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BackgroundThe oriental melon (Cucumis meloL. var.makuwa) is one of the most important cultivated cucurbits grown widely in Korea, Japan, and northern China. It is cultivated because its fruit has a sweet aromatic flavor and is rich in soluble sugars, organic acids, minerals, and vitamins. In order to elucidate the genetic and molecular basis of the developmental changes that determine size, color, and sugar contents of the fruit, we performedde novotranscriptome sequencing to analyze the genes expressed during fruit development.ResultsWe identified a total of 47,666 of representative loci from 100,875 transcripts and functionally annotated 33,963 of the loci based on orthologs inArabidopsis thaliana. Among those loci, we identified 5,173 differentially expressed genes, which were classified into 14 clusters base on the modulation of their expression patterns. The expression patterns suggested that the differentially expressed genes were related to fruit development and maturation through diverse metabolic pathways. Analyses based on gene set enrichment and the pathways described in the Kyoto Encyclopedia of Genes and Genomes suggested that the expression of genes involved in starch and sucrose metabolism and carotenoid biosynthesis were regulated dynamically during fruit development and subsequent maturation.ConclusionOur results provide the gene expression patterns related to different stages of fruit development and maturation in the oriental melon. The expression patterns give clues about important regulatory mechanisms, especially those involving starch, sugar, and carotenoid biosynthesis, in the development of the oriental melon fruit.

It has been shown that acute and chronic alcohol administrations increase the production of reactive oxygen species, lower cellular antioxidant levels and enhance oxidative stress in many tissues. We recently reported that cytosolic NADP(+)-dependent isocitrate dehydrogenase (IDPc) functions as an antioxidant enzyme by supplying NADPH to the cytosol. Upon exposure to ethanol, IDPc was susceptible to the loss of its enzyme activity in HepG2 cells. Transfection of HepG2 cells with an IDPc small interfering RNA noticeably downregulated IDPc and enhanced the cells' vulnerability to ethanol-induced cytotoxicity. Our results suggest that suppressing the expression of IDPc enhances ethanol-induced toxicity in HepG2 cells by further disruption of the cellular redox status.

Myeoloperoxidase catalyses the formation of hypochlorous acid (HOCl) via reaction of H(2)O(2) with Cl(-) ion. Although HOCl is known to play a major role in the human immune system by killing bacteria and other invading pathogens, excessive generation of this oxidant is known to cause damage to tissue. Recently, it was demonstrated that the control of mitochondrial redox balance and oxidative damage is one of the primary functions of mitochondrial NADP(+)-dependent isocitrate dehydrogenase (IDPm) to supply NADPH for antioxidant systems. This study investigated whether the IDPm would be a vulnerable target of HOCl as a purified enzyme and in intact cells. Loss of enzyme activity was observed and the inactivation of IDPm was reversed by thiols. Transfection of HeLa cells with an IDPm small interfering RNA (siRNA) markedly enhanced HOCl-induced oxidative damage to cells. The HOCl-mediated damage to IDPm may result in the perturbation of the cellular antioxidant defense mechanisms and subsequently lead to a pro-oxidant condition.

Singlet oxygen is a highly reactive form of molecular oxygen that may harm living systems by oxidizing critical cellular macromolecules and it also promotes deleterious processes such as cell death. Recently, we demonstrated that the control of redox balance and the cellular defense against oxidative damage are the primary functions of cytosolic NADP(+)-dependent isocitrate dehydrogenase (IDPc) through supplying NADPH for antioxidant systems. In this report, we demonstrate that modulation of IDPc activity in HL-60 cells regulates singlet oxygen-induced apoptosis. When we examined the protective role of IDPc against singlet oxygen-induced apoptosis with HL-60 cells transfected with the cDNA for mouse IDPc in sense and antisense orientations, a clear inverse relationship was observed between the amount of IDPc expressed in target cells and their susceptibility to apoptosis. The results suggest that IDPc plays an important protective role in apoptosis of HL-60 cells induced by singlet oxygen.

Purpose Inflammation-based prognostic scores including neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and lymphocyte-to-monocyte ratio (LMR) are associated with oncologic outcomes in diverse malignancies. We evaluated the predictive value of pretreatment prognostic scores in differentiating nonmuscle invasive (NMIBC) and muscle invasive bladder cancer (MIBC). Materials and Methods Consecutive transurethral resection of bladder tumour (TURBT) cases from January 2011 to December 2013 were analysed retrospectively. Patient demographics, tumour characteristics and prognostic scores results were recorded. Receiver operating characteristics curves were used to determine prognostic score cutoffs. Univariate and multivariate binomial logistic regression analysis was performed to evaluate the association between variables and MIBC. Results A total of 226 patients were included, with 175 and 51 having NMIBC (stages Ta and T1) and MIBC (stage T2+) groups, respectively. Median age was 75 years and 174 patients were male. The NLR cutoff was 3.89 and had the greatest area under the curve (AUC) of 0.710, followed by LMR (cutoff218; AUC, 0.642). Full blood count samples were taken a median of 12 days prior to TURBT surgery. Multivariate logistic regression analysis identified tumour grade G3 (odds ration [OR], 32.848; 95% confidence interval [CI], 9.818-109.902; p=0.000), tumour size≥3 cm (OR, 3.353; 95% CI, 1.347-8.345; p=0.009) and NLR≥3.89 (OR, 8.244; 95% CI, 2.488-27.316; p=0.001) as independent predictors of MIBC. Conclusions NLR may provide a simple, cost-effective and easily measured marker for MIBC. It can be performed at the time of diagnostic flexible cystoscopy, thereby assisting in the planning of further treatment.

NADPH is an important cofactor in many biosynthesis pathways and the regeneration of reduced glutathione, critically important in cellular defense against oxidative damage. It is mainly produced by glucose 6-phosphate dehydrogenase (G6PD), malic enzyme, and the cytosolic form of NADP(+)-dependent isocitrate dehydrogenase (IDPc). Little information is available about the role of IDPc in antioxidant defense. In this study we investigated the role of IDPc against cytotoxicity induced by oxidative stress by comparing the relative degree of cellular responses in three different NIH3T3 cells with stable transfection with the cDNA for mouse IDPc in sense and antisense orientations, where IDPc activities were 3-4-fold higher and 35% lower, respectively, than that in the parental cells carrying the vector alone. Although the activities of other antioxidant enzymes, such as superoxide dismutase, catalase, glutathione reductase, glutathione peroxidase, and G6PD, were comparable in all transformed cells, the ratio of GSSG to total glutathione was significantly higher in the cells expressing the lower level of IDPc. This finding indicates that IDPc is essential for the efficient glutathione recycling. Upon transient exposure to increasing concentrations of H(2)O(2) or menadione, an intracellular source of free radicals and reactive oxygen species, the cells with low levels of IDPc became more sensitive to oxidative damage by H(2)O(2) or menadione. Lipid peroxidation, oxidative DNA damage, and intracellular peroxide generation were higher in the cell-line expressing the lower level of IDPc. However, the cells with the highly over-expressed IDPc exhibited enhanced resistance against oxidative stress, compared to the control cells. This study provides direct evidence correlating the activities of IDPc and the maintenance of the cellular redox state, suggesting that IDPc plays an important role in cellular defense against oxidative stress.

Recently, we demonstrated that the control of cytosolic and mitochondrial redox balance and the cellular defense against oxidative damage is one of the primary functions of NADP+-dependent isocitrate dehydrogenase (ICDH) through supply of NADPH for antioxidant systems. When exposed to various reactive oxygen species such as hydrogen peroxide, singlet oxygen generated by photoactivated dye, superoxide anion, and hydroxyl radical produced by metal-catalyzed Fenton reactions, ICDH was susceptible to oxidative modification and damage, which was indicated by the loss of activity, fragmentation of the peptide as well as by the formation of carbonyl groups. Oxidative damage to ICDH was inhibited by antioxidant enzymes, free radical scavengers, and spin-trapping agents. The structural alterations of modified enzymes were indicated by the increase in thermal instability and binding of the hydrophobic probe 8-anilino-1-naphthalene sulfonic acid (ANSA). The reactive oxygen species-mediated damage to ICDH may result in the perturbation of cellular antioxidant defense mechanisms and subsequently lead to a pro-oxidant condition.

A soluble protein from Saccharomyces cerevisiae specifically provides protection against a thiol-containing oxidation system but not against an oxidation system without thiol. This 25-kDa protein acts as a peroxidase but requires a NADPH-dependent thioredoxin system or a thiol-containing intermediate, and was thus named thioredoxin peroxidase (TPx). The protective role of TPx in the cellular defense against heat shock (42 or 48 degreesC), which may increase oxidative stress in cells sufficiently to form reactive oxygen species harmful to cellular function, was investigated in a wild-type and a mutant yeast strain in which the tsa gene that encodes TPx was disrupted by homologous recombination. Upon exposure under aerobic conditions to heat shock there was a distinct difference between these two strains in growth kinetics and viability. The wild-type strain was more resistant to killing by heat than the mutant strain. In addition, the expression of the tsa gene in Escherichia coli caused an increase in thermotolerance. The expression of the tsa gene increased under heat shock; however, modulation of activities of other antioxidant enzymes, such as catalase, superoxide dismutase, glucose 6-phosphate dehydrogenase, and glutathione reductase as well as the total glutathione level, remained unaltered in both strains under heat shock. The induction of heat shock protein HSP104 was not significantly different in the two strains under heat shock. The results indicate that the lack of TPx expression may be solely responsible for the thermosensitive phenotype of tsa mutant cells. When the oxidation of 2', 7'-dichlorofluorescin was used to examine hydroperoxide production in yeast cells, tsa mutant cells showed a 2.5- to 3.5-fold increase in fluorescence upon exposure to heat stress compared to wild-type cells. The antioxidant, N-acetylcysteine, prevented intracellular peroxide formation in response to heat shock. The carbonyl content of extract, the indicative marker of oxidative damage to protein, from tsa mutant cells was higher than that from wild-type cells. These results suggest that TPx may play a direct role in cellular defense against heat shock, presumably functioning as an antioxidant protein.

Superoxide dismutase (SOD) is a key enzyme in the antioxidant system of the cells. When exposed to a metal-catalyzed oxidation (MCO) system composed of Fe3+, O2, and thiol as an electron donor copper, zinc SOD (CuZnSOD) was susceptible to oxidative modification and damage as indicated by the loss of activity, fragmentation and aggregation of peptide as well as by the formation of carbonyl groups. Oxidative damage to CuZnSOD was inhibited by diethylenetriaminepentaacetic acid as well as by free radical scavengers and spin-trapping agents. The results of the present study indicate that hydrogen peroxide may be generated from a thiol/Fe3+/O2 system and that hydroxyl free radicals, produced by metal-catalyzed Fenton reactions, may be the ultimate species mediating the SOD damage. Incubation with the MCO system resulted in the release of Cu ions from CuZnSOD. Incubation with the thiol-MCO did not significantly increase the formation of 2-oxohistidine in CuZnSOD. The lack of formation of 2-oxohistidine, as well as the pronounced preventive effect of spin-traps on the thiol-MCO-mediated damage to CuZnSOD, indicates that inactivation might actually be predominantly due to global oxidation rather than a site-specific oxidation. The thiol-MCO-mediated damage to SOD may result in the perturbation of cellular antioxidant defense mechanisms and subsequently lead to a pro-oxidant condition.

Oxidative DNA damage caused by hydrogen peroxide was enhanced by copper, zinc superoxide dismutase (CuZnSOD) in a concentration-dependent manner, as reflected by the formation of 8-hydroxy-2'-deoxyguanosine (8-OH-dG) and strand breaks. Hydroxyl radical scavengers such as sodium azide, mannitol and 5,5-dimethyl-1-pyrroline N-oxide (DMPO), a metal chelator, diethylenetriamine-pentaacetic acid, and catalase decreased strand breaks and 8-OH-dG formation in DNA. The deoxyribose assay showed that hydroxyl free radicals were generated in the reaction of CuZnSOD with H2O2. CuZnSOD also caused enhancement of mutation in the pUC18 lacZ' gene in the presence of H2O2 when measured as a loss of alpha-complementation. Based on these results, we interpret the effects of CuZnSOD on hydrogen peroxide induced DNA damage and mutation as due to reactive oxygen species, probably hydroxyl free radicals, formed predominantly by the reaction of hydrogen peroxide and free Cu2+ released from oxidatively damaged CuZnSOD.

Angular pregnancy refers to implantation of the embryo just medial to uterotubal junction in the lateral angle of the uterine cavity. This is differentiated from interstitial pregnancy in which the gestational sac is located within intramural portion of the tube. Angular pregnancy is categorized as intrauterine pregnancy and the outcome is known favorable. However, there have been few reports about adverse perinatal complications such as abortion, retained placenta, placenta accreta, placenta percreta and uterine rupture. We report a case of angular pregnancy complicated with preterm labor at 25 gestational weeks, placenta accreta and postpartum endometritis. This case suggests that angular pregnancy should be differentiated from normal intrauterine pregnancy because of its potential risk of adverse outcome.

A novel method based on a statistical model for the fundamental-frequency (F0) synthesis in Mandarin text-to-speech is proposed. Specifically, a statistical model is employed to determine the relationship between F0 contour patterns of syllables and linguistic features representing the context. Parameters of the model were empirically estimated from a large training set of sentential utterances. Phonologic rules are then automatically deduced through the training process and implicitly memorized in the model. In the synthesis process, contextual features are extracted from a given input text, and the best estimates of F0 contour patterns of syllable are then found by a Viterbi algorithm using the well-trained model. This method can be regarded as employing a stochastic grammar to reduce the number of candidates of F0 contour pattern at each decision point of synthesis. Although linguistic features on various levels of input text can be incorporated into the model, only some relevant contextual features extracted from neighboring syllables were used in this study. Performance of this method was examined by simulation using a database composed of nine repetitions of 112 declarative sentential utterances of the same text, all spoken by a single speaker. By closely examining the well-trained model, some evidence was found to show that the declination effect as well as several sandhi rules are implicitly contained in the model. Experimental results show that 77.56% of synthesized F0 contours coincide with the VQ-quantized counterpart of the original natural speech. Naturalness of the synthesized speech was confirmed by an informal listening test.

【Fe powders with elongated and aggregated structure as heat pellet material for thermal battery applications were prepared by spray pyrolysis under various preparation conditions. The precursor powders with spherical shapes and hollow morphologies turned into Fe powders after reduction at a temperature of 615 $^{\circ}C$ under 20% $H_2$ /Ar gas. The powders had pure Fe crystal structures irrespective of the preparation conditions of the precursor powders in the spray pyrolysis. The morphologies and mean sizes of the Fe powders are affected by the preparation conditions of the precursor powders in the spray pyrolysis. Therefore, the ignition sensitivities and the burn rates of the heat pellets formed from the Fe powders prepared by spray pyrolysis are affected by the preparations of the precursor powders. The Fe powders prepared under the optimum preparation conditions have a BET surface area of 2.9 $m^2g^1$ . The heat pellets prepared from the Fe powders with elongated and aggregated structure have a good ignition sensitivity of 1.1W and a high burn rate of 18 $cms^1$ .】

INK4a and human telomerase reverse transcriptase (hTERT ) genes and the p16 and hTERT protein expressions in invasive ductal carcinoma of the breast. In addition, we evaluated the relationship between the methylation status of the two genes and their protein expressions. Methods: We performed methylation-specific PCR (MSP) and immunohistochemical staining in 63 breast cancer specimens. Results: There was no statistical association between p16 INK4a gene methylation and the histological grade (tumor grade, tumor size and lymph node status). Methylation of the hTERT promoter did show significant differences according to the histological tumor grade and tumor size, but there was no clinical significance. Methylation of the p16 INK4a and hTERT genes was found in 22.2% and 31.8% of the specimens, respectively. A negative p16 protein expression (0-10% expression rate) was observed in 38.1% of the specimens (24 of 63). A positive hTERT expression (more than a 25% expression rate) was observed in 73.0% of the specimens (46 of 63). There was no statistical significance in the relationship between the methylation status and the protein expression. Conclusion: Our data suggest that methylation of the p16 and hTERT genes is not associated with their protein expressions according to Immunohistochemisty. There seemed to be another complicated mechanism for p16 inactivation and hTERT activation in breast cancer. (J Breast Cancer 2007; 10:59-67)

Major depressive disorder is associated with impairments in processing emotional stimuli, and residual impairments are observed during remission, possibly indicating trait vulnerability. Stimuli with social context represent a distinct class of emotional stimuli, which in healthy volunteers are associated with specific neural substrates but have not previously been studied relative to vulnerability to depression.To explore whether individuals with remitted major depressive disorder had altered neuronal processing of social emotional stimuli.Cross-sectional design using functional magnetic resonance imaging, combined with a cognitive activation task.General community of greater Manchester, England.Twenty-five unmedicated participants fully remitted from major depressive disorder and 29 age-matched control subjects.Neuronal responses to positive and negative social interaction images vs valence-matched images with less overt social context.Participants with remitted depression showed attenuated frontopolar response relative to controls for positive and negative images depicting social interactions. For negative social images, participants with remitted depression also showed reduced latero-orbitofrontal response relative to controls.In the absence of current symptoms, individuals with remitted major depressive disorder showed reduced frontopolar processing of stimuli showing social interactions, a reduction not seen for stimuli showing individual successes and failures and, therefore, not simply an effect of emotional valence. These results suggest a specific trait abnormality in social emotional processing associated with vulnerability to depression, which may have implications for understanding social cognition mechanisms and for developing effective psychological therapies.

Background Active ‘hands-on’ participation in the ‘hot-seat’ during immersive simulation-based training (SBT) induces stress for participants, which is believed to be necessary to improve performance. We hypothesized that observers of SBT can subsequently achieve an equivalent level of non-technical performance as ‘hot-seat’ participants despite experiencing lower stress. Methods We randomized 37 anaesthesia trainees into two groups to undergo three consecutive SBT scenarios. Eighteen ‘hot-seat’ trainees actively participated in all three scenarios, and 19 ‘observer’ trainees were directed to observe the first two scenarios and participated in the ‘hot-seat’ only in scenario 3. Salivary cortisol (SC) was measured at four time points during each scenario. Primary endpoint for stress response was the change in SC (ΔSC) from baseline. Performance was measured using the Anaesthetist’s Non-Technical Skills (ANTS) Score. Results Mean SC increased in all participants whenever they were in the ‘hot-seat’ role, but not when in the observer role. Hot-seat ΔSC (mcg/dL) for scenarios 1, 2, and 3 were 0.122 (p = 0.001), 0.074 (p = 0.047), and 0.085 (p = 0.023), respectively. Observers ΔSC (mcg/dL) for scenarios 1, 2, and 3 were −0.062 (p = 0.091), 0.010 (p = 0.780), and 0.144 (p = 0.001), respectively. Mean ANTS scores were equivalent between the ‘hot-seat’ (40.0) and ‘observer’ (39.4) groups in scenario 3 (p = 0.733). Conclusions Observers of SBT achieved an equivalent level of non-technical performance, while experiencing lower stress than trainees repeatedly trained in the ‘hot-seat’. Our findings suggest that directed observers may benefit from immersive SBT even without repeated ‘hands-on’ experience and stress in the hot-seat. The directed observer role may offer a less stressful, practical alternative to the traditional ‘hot-seat’ role, potentially rendering SBT accessible to a wider audience. Trial registration ClinicalTrials.gov Identifier NCT02211378, registered August 5, 2014, retrospectively registered. Electronic supplementary material The online version of this article (doi:10.1186/s41077-017-0040-7) contains supplementary material, which is available to authorized users.