A genome-wide association study identifies two novel susceptibility loci and trans population polygenicity associated with bipolar disorder.

Genome-wide association studies (GWASs) have identified several susceptibility loci for bipolar disorder (BD) and shown that the genetic architecture of BD can be explained by polygenicity, with numerous variants contributing to BD. In the present GWAS (Phase I/II), which included 2964 BD and 61 887 control subjects from the Japanese population, we detected a novel susceptibility locus at 11q12.2 (rs28456, P=6.4 × 10-9), a region known to contain regulatory genes for plasma lipid levels (FADS1/2/3). A subsequent meta-analysis of Phase I/II and the Psychiatric GWAS Consortium for BD (PGC-BD) identified another novel BD gene, NFIX (Pbest=5.8 × 10-10), and supported three regions previously implicated in BD susceptibility: MAD1L1 (Pbest=1.9 × 10-9), TRANK1 (Pbest=2.1 × 10-9) and ODZ4 (Pbest=3.3 × 10-9). Polygenicity of BD within Japanese and trans-European-Japanese populations was assessed with risk profile score analysis. We detected higher scores in BD cases both within (Phase I/II) and across populations (Phase I/II and PGC-BD). These were defined by (1) Phase II as discovery and Phase I as target, or vice versa (for 'within Japanese comparisons', Pbest~10-29, R2~2%), and (2) European PGC-BD as discovery and Japanese BD (Phase I/II) as target (for 'trans-European-Japanese comparison,' Pbest~10-13, R2~0.27%). This 'trans population' effect was supported by estimation of the genetic correlation using the effect size based on each population (liability estimates~0.7). These results indicate that (1) two novel and three previously implicated loci are significantly associated with BD and that (2) BD 'risk' effect are shared between Japanese and European populations.

What is the primary mover of water dynamics?

Even today, the H-bonded cluster structure of water still stands as a major point of debate in the science of liquids. Much of this discussion is devoted to understand its dynamic nature. This has a direct impact on deciphering the many anomalies of water such as its exceptional heat capacity. Of these properties, dielectric permittivity and relaxation are of particular interest. The argument rages over whether the almost Debye-like character of the dispersion is the result of the reorientation of an apparent dipole moment of the water cluster or simply the cumulative effect of single water molecule reorientation. Furthermore, like many glass formers, it has a high frequency excess wing that does not fit into the accepted models of a single relaxation time of the main peak. Herein, we present evidence that the microscopic origins of both the excess wing and the main relaxation process of pure water are the same. The origin of these two features is explored and we suggest a new paradigm for water relaxation based on the concept of a proton cascade leading to a cluster reorientation.

Work-life balance of female versus male surgeons in Hong Kong based on findings of a questionnaire designed by a Japanese surgeon.

PURPOSE: We evaluated the attitudes of female and male surgeons in Hong Kong to work, personal life, and work-life balance, through a questionnaire survey. METHODS: We sent an online questionnaire survey designed by a female Japanese surgeon to 142 female surgeons practicing in Hong Kong, while its customized version was sent to 953 of their male counterparts. RESULTS: "Home life" and "Work" were the first and second priorities for both the women and the men. More of the female surgeons reported that they could not find enough time to participate in community activities (p = 0.038) or rest (p = 0.024). Both reported moderate satisfaction at work (p = 0.114) and in their life outside work (p = 0.346), as well as equality at home (p = 0.548) and at work (p = 0.177). Of the men, 89 % agreed with the necessity for granting paternity leave and reported that they would like to work part-time during the child-rearing years (p = 0.013). CONCLUSIONS: The number of female surgeons involved in key roles is increasing in parallel with the increasing number of female medical students. The introduction of paternity leave for male surgeons is an important concern. The issues of work-life balance may need to be addressed to attract more talent into an important specialty in Hong Kong and this is likely to apply to other parts of Asia as well.

Micronized benzoic acid decreases the concentration necessary to preserve acidic beverages against Alicyclobacillus.

AIMS: The aim of this study was a challenge testing the effect of lower concentrations of micronized benzoic acid against two strains of Alicyclobacillus. METHODS AND RESULTS: The effect of micronized benzoic acid was compared with the usual levels of untreated commercial sodium benzoate and benzoic acid, at the challenge temperature of 45°C. The size of the benzoic acid particles was determined by scanning electron microscopy. The diameter of the micronized particles was around 10 µm with a maximum length of 200 µm, while the untreated preservative structures were irregular with lengths up to 500 µm. A continuous bactericidal effect against two Alicyclobacillus strains, throughout the 28-day period, was observed with 50 mg l(-1) of micronized benzoic acid, but when the untreated preservative was used, the same lethal effect was not achieved even after doubling its concentration. CONCLUSIONS: The antimicrobial activity of benzoic acid was improved by micronization. The process proved to be an effective alternative to reduce the benzoic acid concentration necessary to ensure stability of an orange juice matrix. SIGNIFICANCE AND IMPACT OF THE STUDY: The results proved that the micronization process represents an alternative to reduce the required food preservative concentration; this method increased the stability of the compound, which maintains its bioavailability.

Soft-x-ray harmonic comb from relativistic electron spikes.

We demonstrate a new high-order harmonic generation mechanism reaching the "water window" spectral region in experiments with multiterawatt femtosecond lasers irradiating gas jets. A few hundred harmonic orders are resolved, giving µJ/sr pulses. Harmonics are collectively emitted by an oscillating electron spike formed at the joint of the boundaries of a cavity and bow wave created by a relativistically self-focusing laser in underdense plasma. The spike sharpness and stability are explained by catastrophe theory. The mechanism is corroborated by particle-in-cell simulations.

Enhancement of photon number reflected by the relativistic flying mirror.

Laser light reflection by a relativistically moving electron density modulation (flying mirror) in a wake wave generated in a plasma by a high intensity laser pulse is investigated experimentally. A counterpropagating laser pulse is reflected and upshifted in frequency with a multiplication factor of 37-66, corresponding to the extreme ultraviolet wavelength. The demonstrated flying mirror reflectivity (from 3 x 10(-6) to 2 x 10(-5), and from 1.3 x 10(-4) to 0.6 x 10(-3), for the photon number and pulse energy, respectively) is close to the theoretical estimate for the parameters of the experiment.

Electron optical injection with head-on and countercrossing colliding laser pulses.

A high stability electron bunch is generated by laser wakefield acceleration with the help of a colliding laser pulse. The wakefield is generated by a laser pulse; the second laser pulse collides with the first pulse at 180 degrees and at 135 degrees realizing optical injection of an electron bunch. The electron bunch has high stability and high reproducibility compared with single pulse electron generation. In the case of 180 degrees collision, special measures have been taken to prevent damage. In the case of 135 degrees collision, since the second pulse is countercrossing, it cannot damage the laser system.

Mechanism of ASC-mediated apoptosis: bid-dependent apoptosis in type II cells.

Apoptosis-associated speck-like protein containing a CARD (ASC) is an adaptor molecule that mediates apoptotic and inflammatory signals, and implicated in tumor suppression. However, the mechanism of ASC-mediated apoptosis has not been well elucidated. Here, we investigated the molecular mechanisms of ASC-mediated apoptosis in several cell lines using a caspase recruitment domain 12-Nod2 chimeric protein that transduces the signal from muramyl dipeptide into ASC-mediated apoptosis. Experiments using dominant-negative mutants, small-interfering RNAs and peptide inhibitors for caspases indicated that caspase-8 was generally required for ASC-mediated apoptosis, whereas a requirement for caspase-9 depended on the cell type. In addition, caspase-like apoptosis-regulatory protein (CLARP)/Fas-like inhibitor protein, a natural caspase-8 inhibitor, suppressed ASC-mediated apoptosis, and Clarp-/- mouse embryonic fibroblasts were highly sensitive to ASC-mediated apoptosis. Bax-deficient HCT116 cells were resistant to ASC-mediated apoptosis as reported previously, although we failed to observe colocalization of ASC and Bax in cells. Like Fas-ligand-induced apoptosis, the ASC-mediated apoptosis was inhibited by Bcl-2 and/or Bcl-XL in type-II but not type-I cell lines. Bid was cleaved upon ASC activation, and suppression of endogenous Bid expression using small-interfering RNAs in type-II cells reduced the ASC-mediated apoptosis. These results indicate that ASC, like death receptors, mediates two types of apoptosis depending on the cell type, in a manner involving caspase-8.

Terahertz spectroscopy of native-conformation and thermally denatured bovine serum albumin (BSA).

Proteins are expected to exhibit collective vibrational modes at terahertz frequencies. We have developed a promising approach to measure these motions by using a membrane device to hold samples. Samples of bovine serum albumin (BSA) in native and thermally denatured conformations were measured using terahertz time-domain spectroscopy. Clear differences were observed in transmittance and phase between native-conformation BSA samples and thermally denatured BSA samples. Time-domain data shows that samples exhibited relative time shifts when compared with a standard. Results suggest that there were differences in dielectric responses in the BSA samples, and these are probably associated with molecular conformational changes in the membrane device.

Sub-MeV tunably polarized X-ray production with laser Thomson backscattering.

Reported in this article is the generation of unique polarized x-rays in the sub-MeV region by means of the Thomson backscattering of the Nd:YAG laser photon with a wavelength of 1064 nm on the 150 MeV electron from the microtron accelerator. The maximum energy of the x-ray photons is estimated to be about 400 keV. The total energy of the backscattered x-ray pulse is measured with an imaging plate and a LYSO scintillator. The angular divergence of the x-rays is also measured by using the imaging plate. We confirm that the x-ray beam is polarized according to the laser polarization direction with the Compton scattering method. In addition, we demonstrate the imaging of the object shielded by lead with the generated x-rays.

The yeast MOT2 gene encodes a putative zinc finger protein that serves as a global negative regulator affecting expression of several categories of genes, including mating-pheromone-responsive genes.

The STE4 gene encodes the beta subunit of a heterotrimeric G protein that is an essential component of the pheromone signal transduction pathway. To identify downstream component(s) of Ste4, we sought pseudo-revertants that restored mating competence to ste4 mutants. The suppressor mot2 was isolated as a recessive mutation that restored conjugational competence to a temperature-sensitive ste4 mutant and simultaneously conferred a temperature-sensitive growth phenotype. The MOT2 gene encodes a putative zinc finger protein, the deletion of which resulted in temperature-sensitive growth, increased expression of FUS1 in the absence of pheromones, and suppression of a deletion of the alpha-factor receptor. On the other hand, sterility resulting from deletion of STE4 was not suppressed by the mot2 deletion. These phenotypes are similar to those associated with temperature-sensitive mutations in CDC36 and CDC39, which are proposed to encode general negative regulators of transcription rather than factors involved in the pheromone response pathway. Deletion of MOT2 also caused increased transcription of unrelated genes such as GAL7 and PHO84. Overexpression of MOT2 suppresses the growth defect of temperature-sensitive mutations in CDC36 and CDC39. These observations suggest that Mot2 functions as a general negative regulator of transcription in the same processes as Cdc36 and Cdc39.

mda-7 gene transfer sensitizes breast carcinoma cells to chemotherapy, biologic therapies and radiotherapy: correlation with expression of bcl-2 family members.

Current therapies used in the treatment of breast cancer are limited by systemic toxicity, rapid drug metabolism and intrinsic and acquired drug resistance. We have previously shown that adenoviral-mediated transfer of the melanoma differentiation-associated gene-7 (mda-7) elicits growth inhibition and apoptosis in various tumor types. Here, we evaluate the effects of Ad-mda7, alone and in combination with other therapies, against a panel of nine breast tumor cell lines and their normal counterparts; we report selective Ad-mda7-mediated p53-independent growth inhibition, G2/M cell cycle arrest, and apoptosis. In vivo, Ad-mda7 induced p53-independent tumor growth inhibition (P<0.004) in multiple xenograft models. We then evaluated the combination of Ad-mda7 with agents commonly used to treat breast cancer: radiotherapy (XRT), Tamoxifen, Taxotere, Adriamycin, and Herceptin. These agents exhibit diverse modes of action, including formation of bulky adducts, inhibition of DNA replication (Adriamycin, XRT), damage to microtubules (Taxotere), nonsteroidal estrogen antagonists (Tamoxifen), or Her2/neu receptor blockade (Herceptin). Treated with conventional anticancer drugs or radiation, MDA-7-expressing cells display additive or synergistic cytotoxicity and apoptosis that correlates with decreased BCL-2 expression and BAX upregulation. In vivo, animals that received Ad-mda7 and XRT underwent significant reduction of tumor growth (P<0.002). This is the first report of the synergistic effects of Ad-mda7 combined with chemotherapy or radiotherapy on human breast carcinoma cells.

Lactoferrin inhibits cholesterol accumulation in macrophages mediated by acetylated or oxidized low-density lipoproteins.

When macrophages are incubated with acetylated or oxidized low-density lipoproteins (Ac- or OxLDL), cellular cholesteryl esters (CE) increase significantly. In the present study, we investigated the effect of whey protein on Ac- or OxLDL mediated accumulation of CE in macrophages and found that lactoferrin (Lf), a minor protein component of whey, inhibits the accumulation of CE dose-dependently. In the presence of bovine Lf (1 mg/ml), CE accumulation in macrophages incubated with AcLDL (100 micrograms of protein/ml) decreased by more than 80%. Human Lf was less potent than bovine Lf, and bovine transferrin had no effect. Binding of 125I-AcLDL to macrophages was also inhibited by Lf. Agarose gel electrophoresis revealed that Lf binds to Ac- or OxLDLs and neutralizes their negative charges. These results indicate that Lf inhibits the binding of modified LDLs to macrophages by direct interaction with modified LDLs, resulting in their loss of function as ligands of the scavenger receptor. Modification of the arginine residues of Lf with 1,2-cyclohexanedione abolished its ability to bind to AcLDL, suggesting that a region rich in basic amino acid residues near the N-terminus of Lf, which resembles the ligand-binding site of the scavenger receptor, may be responsible for this binding ability. As a result, the inhibitory effect of Lf on CE accumulation in macrophages was significantly weakened by this modification. Our results suggest the possibility that Lf in the blood stream may act as an anti-atherogenic agent in vivo.

Identification of the bactericidal domain of lactoferrin.

We report the existence of a previously unknown antimicrobial domain near the N-terminus of lactoferrin in a region distinct from its iron-binding sites. A single active peptide representing this domain was isolated following gastric pepsin cleavage of human lactoferrin, and bovine lactoferrin, and sequenced by automated Edman degradation. The antimicrobial sequence was found to consist mainly of a loop of 18 amino acid residues formed by a disulfide bond between cysteine residues 20 and 37 of human lactoferrin, or 19 and 36 of bovine lactoferrin. Synthetic analogs of this region similarly exhibited potent antibacterial properties. The active peptide of bovine lactoferrin was more potent than that of human lactoferrin having effectiveness against various Gram-negative and Gram-positive bacteria at concentrations between 0.3 microM and 3.0 microM, depending on the target strain. The effect of the isolated domain was lethal causing a rapid loss of colony-forming capability. Our studies suggest this domain is the structural region responsible for the bacterial properties of lactoferrin.

Intervention of maternal transmission of HTLV-1 in Nagasaki, Japan.

Seroepidemiological and laboratory virological evidences strongly suggested that endemicity of HTLV-1 in Nagasaki Japan depends on maternal infant infections via breast milk. The most obvious way to prove this concept was an intervention study with refraining from breast-feeding by carrier mothers. Most infected babies seroconverted by the age of 12 months, which made it possible to diagnose the infection at the age of 12 months for the statistical purpose. Serology and PCR on both adults and children were consistent each other, suggesting the absence of seronegative carriers. The intervention study revealed that approximately 80% of maternal infection was prevented by refraining from breast feeding by carrier mothers. The remaining fraction of infections in formula-fed babies suggested an alternative infection pathway. Although intrauterine infections has been suggested by others to explain the PCR-positive cord blood samples. However, groups of cord blood-positive children and seroconverted children were distinct each other. Therefore, the presence of HTLV-1 provirus in the cord blood can not be a marker of intrauterine infection. Mothers who infected a child has approximately 10 times higher risk of another infection for the next baby than those who did not.

Promoter analysis of RPE65, the gene encoding a 61-kDa retinal pigment epithelium-specific protein.

PURPOSE: To identify the functional promoter region and cis-acting elements that regulate the expression of RPE65, the retinal pigment epithelium (RPE)-specific gene responsible for certain forms of autosomal recessive childhood-onset severe retinal dystrophy. METHODS: A human genomic DNA clone containing the 5'-flanking region of RPE65 was isolated and, 4.0 kb proximal to the transcription start site, was sequenced and analyzed for the presence of transcription factor-binding sites. Promoter activity was assayed by transient transfection of luciferase reporter constructs containing nested deletions of the upstream sequence in the human RPE cell lines ARPE19 and D407, as well as in the SK-Mel-28 and HeLa cell lines. Specific DNA protein-binding sites present in the 340 bp upstream of the transcription start site were identified by DNase I footprint analysis. RESULTS: Sequence analysis places the polymorphic marker, D1S2803, within the RPE65 upstream region and identifies a number of sequences homologous to the gene encoding the cellular retinaldehyde-binding protein. Functional analysis indicates that basal promoter activity is conferred by the sequence from -83 to +39 and is approximately equivalent in all cell lines tested, with no other control elements detected in 3.6 kb of the upstream sequence. At least eight protected regions are identified in DNase I footprint assays, including sequences corresponding to the predicted TATA box, AP-4, and nuclear factor-1 DNA protein-binding sites. CONCLUSIONS: These findings localize the basal promoter activity of RPE65, identify potential cis-acting elements that act as positive regulators of gene expression, and suggest that additional regulatory elements are likely to be involved in restricting gene expression to the retinal pigment epithelium. Identification of promoter elements and genetic markers in the upstream sequence will enable the screening of patients with retinal degeneration for possible mutations that affect RPE65 expression.

A frequency-agile terahertz-wave parametric oscillator.

We demonstrated a pulse-to-pulse frequency-tunable LiNbO3 terahertz-wave parametric oscillator, pumped with a Q-switched Nd:YAG laser. Rapid tuning from 1 to 2 THz, with random frequency accessibility, was achieved by rotating the pump beam angle using an optical beam scanner and a telescope.

Trabeculectomy with mitomycin. A comparative study with fluorouracil.

We conducted a prospective, randomized study to compare mitomycin with fluorouracil for efficacy and safety as an adjunct to trabeculectomy in eyes with glaucoma with poor surgical prognosis. Thirty-two eyes of 32 patients were randomly allocated to either mitomycin (17 eyes) or fluorouracil therapy (15 eyes). Mitomycin (0.2 mg) was applied only once during trabeculectomy, and fluorouracil (5 mg) was subconjunctivally injected 10 times in the 2 weeks after surgery. Fifteen eyes (88%) in the mitomycin group and seven (47%) in the fluorouracil group achieved an intraocular pressure of less than or equal to 20 mm Hg without antiglaucoma medication. The follow-up period was 7 to 12 months. Corneal complications were less common in the group given mitomycin than in that given fluorouracil (12% vs 53%). The incidence of other complications was similar between the two groups.

Effects of prostaglandin D2, lipoxins and leukotrienes on sleep and brain temperature of rats.

Prostaglandin (PG) D2 and four lipoxygenase-derived eicosanoids [lipoxins (LX) A4 and B4, and leukotrienes (LT) C4 and D4] were examined for their effects on sleep and brain temperature in freely-behaving rats. In the first series of experiments, PGD2 was infused into the third ventricle at four different locations between 23:00 and 05:00. In a location apposed to the medial preoptic area (MPO), PGD2 at doses 1, 10 and 100 pmol/min, increased the slow wave sleep (SWS) by 23% (p < or = 0.01), 35% (p < or = 0.05) and 44% (p < or = 0.01), respectively, during the infusion period. In the second series of experiments, LXs and LTs were infused at the location apposed to MPO. Significant increases in SWS were detected with LXA4 at 100 pmol/min (14%, p < or = 0.05), LXB4 at 100 pmol/min (20%, p < or = 0.05), and LTD at 10 pmol/min (17%, p < or = 0.05). An increase in paradoxical sleep (PS) was produced by PGD2 at 1 and 10 pmol/min infusion (p < or = 0.05), but not by any of the lipoxygenase-derived eicosanoids examined. PGD2 also elevated the mean brain temperature during infusion by 0.2 degrees C and 0.9 degrees C at infusion doses 10 and 100 pmol/min, respectively. But PGD2 infusion at 1 pmol/min did not elevate the brain temperature. LXs (excluding LXB4 at 100 pmol/min) and LTs did not alter the brain temperature significantly at the tested doses. We conclude that PGD2 is the most effective sleep promoter among the eicosanoids examined so far.

Intravenous administration of inorganic selenium compounds, inhibitors of prostaglandin D synthase, inhibits sleep in freely moving rats.

Prostaglandin (PG) D2 has been postulated to be an endogenous sleep-promoting factor. Biosynthesis of PGD2 is catalyzed by PGD synthase (prostaglandin-H2 D-isomerase, EC 5.3.99.2), the activity of which is inhibited by inorganic selenium compounds such as SeCl4 and Na2SeO3. We recently examined the effect of intracerebroventricular administration of these selenium compounds on sleep in rats, and demonstrated time- and dose-dependent sleep inhibition. To establish whether this effect of selenium is also produced when the compound is administered systemically, we devised a procedure for intravenous catheterization and examined the effect of these selenocompounds on sleep-wake activity in freely moving rats (n = 35). Each test compound was administered into the inferior vena cava continuously between 11.00 and 17.00 h on the experimental day. SeCl4 time- and dose-dependently inhibited sleep at infusion rates of 5, 7.5, 10 and 20 nmol/microliters per min. During the SeCl4 infusion at 20 nmol/microliters per min, slow-wave sleep and paradoxical sleep were reduced to 63% and 50% of their respective baseline values. Na2SeO3 exhibited a similar sleep inhibition, though Na2SO3 was ineffective. Infusion of SeCl4 at 10 nmol/microliters per min or below produced no consistent changes in the mean brain temperature, or food and water intake during the infusion period. During the nocturnal period subsequent to SeCl4 infusion, sleep was increased by a rebound phenomenon, while a decrease in brain temperature and inhibition of food and water intake dose-dependently occurred. We conclude that systemic administration of these PGD synthase inhibitors has a sleep-reducing potency.