How does a topological inversion change the evolutionary constraints on membrane proteins?

The members of the aquaporin family and those of the ClC chloride ion channel family consist of two-fold tandem repeats. The orientation of the N-terminal domain against membrane is opposite to that of the C-terminal domain. Several lines of evidence suggest that the extracellular and the cytoplasmic environments impose different evolutionary constraints on proteins (e.g. positive-inside rule). Therefore, the different constraints would affect the corresponding regions of the two domains, which are exposed to the different environments. To examine this hypothesis, the N- and the C-terminal domains were aligned and the difference in residue composition or conservation pattern between the two domains was calculated at each alignment site by several methods. Then, the residues corresponding to the sites exhibiting significant difference were mapped onto the tertiary structure. In spite of the difference in the methods, the mapped residues clustered on the pore surface of the channel; in contrast, the number of the residues mapped on the extracellular or cytoplasmic sides of the proteins was small. A minor modification of the methods improved the sensitivity to detect sites related to the positive-inside rule. The results support our hypothesis about the relationship between the topological inversion and the different constraints.     

Variations of emotional display rules within and across cultures: A comparison between Canada, USA, and Japan.

This study investigates emotional display rules for seven basic emotions. The main goal was to compare emotional display rules of Canadians, US Americans, and Japanese across as well as within cultures regarding the specific emotion, the type of interaction partner, and gender. A total of 835 university students participated in the study. The results indicate that Japanese display rules permit the expression of powerful (anger, contempt, and disgust) significantly less than those of the two North American samples. Japanese also think that they should express positive emotions (happiness, surprise) significantly less than the Canadian sample. Furthermore, Japanese varied the display rules for different interaction partners more than the two North American samples did only for powerful emotions. Gender differences were similar across all three cultural groups. Men expressed powerful emotions more than women and women expressed powerless emotions (sadness, fear) and happiness more than men. Depending on the type of emotion and interaction partner some shared display rules occurred across culture and gender. The implications of these findings are discussed in relation to cultural dimensions and other cultural characteristics. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Possible Action of Olaparib for Preventing Invasion of Oral Squamous Cell Carcinoma In Vitro and In Vivo

Despite recent advances in treatment, the prognosis of oral cancer remains poor, and prevention of recurrence and metastasis is critical. Olaparib is a PARP1 inhibitor that blocks polyADP-ribosylation, which is involved in the epithelial–mesenchymal transition (EMT) characteristic of tumor recurrence. We explored the potential of olaparib in inhibiting cancer invasion in oral carcinoma using three oral cancer cell lines, HSC-2, Ca9-22, and SAS. Olaparib treatment markedly reduced their proliferation, migration, invasion, and adhesion. Furthermore, qRT-PCR revealed that olaparib inhibited the mRNA expression of markers associated with tumorigenesis and EMT, notably Ki67, Vimentin, β-catenin, MMP2, MMP9, p53, and integrin α2 and β1, while E-Cadherin was upregulated. In vivo analysis of tumor xenografts generated by injection of HSC-2 cells into the masseter muscles of mice demonstrated significant inhibition of tumorigenesis and bone invasion by olaparib compared with the control. This was associated with reduced expression of proteins involved in osteoclastogenesis, RANK and RANKL. Moreover, SNAIL and PARP1 were downregulated, while E-cadherin was increased, indicating the effect of olaparib on proteins associated with EMT in this model. Taken together, these findings confirm the effects of olaparib on EMT and bone invasion in oral carcinoma and suggest a new therapeutic strategy for this disease.     

Pressure-assisted capillary electrophoresis electrospray ionization mass spectrometry for analysis of multivalent anions

We describe a method, based on pressure-assisted capillary electrophoresis coupled to electrospray ionization mass spectrometry (PACE/ESI-MS), that allows the simultaneous and quantitative analysis of multivalent anions, such as citrate isomers, nucleotides, nicotinamide-adenine dinucleotides, and flavin adenine dinucleotide, and coenzyme A (CoA) compounds. Key to the analysis was using a noncharged polymer, poly(dimethylsiloxane), coated to the inner surface of the capillary to prevent anionic species from adsorbing onto the capillary wall. It was also necessary to drive a constant liquid flow toward the MS by applying air pressure to the inlet capillary during electrophoresis to maintain a conductive liquid junction between the capillary and the electrospray needle. Although theoretical plates were inferior to those obtained by CE/ESI-MS using a cationic polymer-coated capillary, the PACE/ESI-MS method improved reproducibility and sensitivity of these anions. Eighteen anions were separated by PACE and selectively detected by a quadrupole mass spectrometer with a sheath-flow electrospray ionization interface. The relative standard deviations (n = 6) of the method were better than 0.6% for migration times and between 1.4% and 6.2% for peak areas. The detection limits for these species were between 0.4 and 3.7 micromol/L with pressure injection of 50 mbar for 30 s (30 nL), that is, mass detection limits calculated in the range from 12 to 110 fmol at a signal-to-noise ratio of 3. The utility of the method was demonstrated by analysis of citrate isomers, nucleotides, dinucleotides, and CoA compounds extracted from Bacillus subtilis cells.     

Synergetic Effects of PARP Inhibitor AZD2281 and Cisplatin in Oral Squamous Cell Carcinoma in Vitro and in Vivo

Cisplatin is a commonly used chemotherapeutic drug for treatment of oral carcinoma, and combinatorial effects are expected to exert greater therapeutic efficacy compared with monotherapy. Poly(ADP-ribosyl)ation is reported to be involved in a variety of cellular processes, such as DNA repair, cell death, telomere regulation, and genomic stability. Based on these properties, poly(ADP-ribose) polymerase (PARP) inhibitors are used for treatment of cancers, such as BRCA1/2 mutated breast and ovarian cancers, or certain solid cancers in combination with anti-cancer drugs. However, the effects on oral cancer have not been fully evaluated. In this study, we examined the effects of PARP inhibitor on the survival of human oral cancer cells in vitro and xenografted tumors derived from human oral cancer cells in vivo. In vitro effects were assessed by microculture tetrazolium and survival assays. The PARP inhibitor AZD2281 (olaparib) showed synergetic effects with cisplatin in a dose-dependent manner. Combinatorial treatment with cisplatin and AZD2281 significantly inhibited xenografted tumor growth compared with single treatment of cisplatin or AZD2281. Histopathological analysis revealed that cisplatin and AZD2281 increased TUNEL-positive cells and decreased Ki67- and CD31-positive cells. These results suggest that PARP inhibitors have the potential to improve therapeutic strategies for oral cancer.     

Consumer Acceptability of Muffins with Flaxseed (Linum usitatissimum)

Flaxseed has recently gained attention as a functional food. The effect of adding flaxseed (7.3%, 11.6%, and 15.5%) to muffins on consumer acceptability was evaluated by a focus group, hedonic scoring, and food action rating scale (FACT). The majority of consumers (82.6%, P < 0.001) rated the control muffin (0 % flaxseed) higher than the flax muffin for appearance, color, flavor, texture, overall acceptability, and food acceptance. Flaxseed muffin (11.6%) was “neither liked nor disliked” to “liked slightly” in overall acceptability. This may have been accounted for by the observation that many persons aged 18 to 25 y did not think they needed to consume “health foods”. This “indifferent” and in some cases “negative” attitude toward the flax muffins, which they considered a “health food”, may have influenced the ratings the flax muffins received. Texture and flavor were strongly and positively correlated (P < 0.01) to overall acceptability. A flaxseed muffin provided at least 16% fiber of the daily recommended value.     

Crystallization of fully hydrogenated and interesterified fat and vegetable oil

Owing to public concern regarding the adverse health effects of trans fatty acids, an alternative technology to trans fats has recently become an important issue. The interesterification of fully hydrogenated vegetable oil and liquid oil blends is one of the most versatile options. This paper reports a physical analysis of high-melting fat (HMF) prepared through the interesterification of fully hydrogenated soybean oil and regular soybean oil, and through fractionation. The thermal and structural properties of the HMF blended with salad oil at a mass ratio of 4:1 (called the HMF blend, hereafter), which was prepared as a model fat blend for margarine, were assessed using X-ray diffraction (XRD), differential scanning calorimetry (DSC), and polarized light microscopy (POM). To observe the polymorphic transformation, all samples were aged after crystallization, and the development of granular crystals during the aging process was observed. We found that the granular crystals are made of SOS/SSO, POS/PSO, and (SOS+POS)/(SSO+PSO) molecular compounds, all of which easily transform into β form with a double-chain-length structure.     

Quantification of amygdalin,prunasin, total cyanide and free cyanide in powdered loquat seeds

ABSTRACT

Amygdalin, prunasin, total cyanide and free cyanide concentrations in 12 powdered loquat seeds were investigated. Loquat (Eriobotrya japonica) is a species of flowering plant in the family Rosaceae, and its fruit is quite popular in Japan. Amygdalin and prunasin were measured using LC–MS/MS. Total cyanide was measured by enzymatic treatment, steam distillation and colorimetric quantification using the pyridine–pyrazolone method. Free cyanide was measured without enzymatic treatment. The mean concentrations of amygdalin, prunasin, total cyanide and free cyanide in powdered loquat seeds were 5900, 760, 410 and 44 mg/kg, respectively. The range of each quantitative value was extensive. Seven out of twelve samples were at risk for exceeding the acute reference dose (ARfD) of cyanide.     

Preparation and critical current density of hot pressed Bi2Sr2Ca2Cu3Ox/Ag tapes

Bi2Sr2Ca2Cu3Ox(Bi-2223)/Ag tapes have been prepared by hot pressing performed at 800–840 °C for 12–96 h under 6 or 12 MPa in air. The highest transport critical current density, Jc, is 3900 A cm–2 at 77 K and 8800 A cm–2 at 65 K under zero magnetic field, which is observed in tape hot pressed at 820 °C under 12 MPa for 24 h twice. The tape has undergone a cold pressing under 260 MPa between hot pressings. Jc is limited to 120 A cm–2 in tape hot pressed for 48 h continuously, in spite of total hot pressing time, temperature and pressure all being the same as for tape hot pressed for 24 h twice. It is found that alternate hot pressing and cold pressing is effective in the preparation of Bi-2223 tapes with high Jc, which is determined by the strength of grain coupling. Grain coupling is strengthened in tape hot pressed with an intermediate cold pressing. © 1998 Chapman & Hall     

Effect of the crystallinity of kaolinite precursors on the properties of mesoporous silicas

Mesoporous silicas (MesoPSs) were hydrothermally synthesized from calcined and selectively acid-leached kaolinites with a range of crystallinity, using cetyltrimethyl ammonium bromide (CTABr), to investigate the effect of the kaolinite crystallinity on the porous properties of the resulting MesoPSs. Four kaolinites were used, with Hinckley indices ranging from 0.51 to 1.20 and (001) crystallite sizes ranging from 20 to 37 nm. After calcination at 600 °C for 24 h they were selectively leached with 2.5 M H₂SO₄ at 90 °C for 2 h to prepare microporous silica (MicroPSs). The Si/Al ratios of these MicroPSs varied from 21 to 82 and their specific surface areas (S_BET) ranged from 169 to 370 m²/g, these parameters tending to increase with decreasing Hinckley index of the kaolinite. MesoPSs were synthesized by reacting the resulting MicroPSs with CTABr in NaOH solution under hydrothermal conditions. The MicroPS was mixed with CTABr, NaOH and water in the molar ratio (MicroPS):CTABr:NaOH:H₂O = 1:0.1:0.3:150. The synthesis was carried out by stirring the suspension at room temperature for 24 h, aging for 24 h, hydrothermal treatment at 110 °C for 24 h and calcination at 560 °C for 6 h to remove the surfactants. The S_BET values of the resulting MesoPSs ranged from 932 to 1240 m²/g, correlating with the S_BET values of the precursor MicroPS and the crystallinity of the kaolinite starting materials.