Biochemical composition,protein quality and amino acid profile of red-toothed triggerfish (Odonus niger), an underutilised fish

The present study aims to assess the seasonal and sex-wise variations in proximate composition, amino acid profile and protein quality of Odonus niger, which will be the first report for this species. High-protein (15.15%–20.05%) and low-fat contents (0.46%–1.94%) were observed for the species studied. The highest protein was observed in September and December for male and female fishes respectively. The total essential amino acid (TEAA) content was 58.65–77.41 mg g⁻¹ (males) and 60.25–77.38 mg g⁻¹ (females) and leucine was the predominant EAA in both the sexes. The EAA index and biological value ranged between 17.50–22.56 and 7.38–12.89 for males and 18.02–22.48 and 7.94–12.80 for female fishes respectively. In male, TEAA followed Sept>Jan > Dec > Aug > Oct > Nov and in female it was Oct > Sept>Dec > Nov > Aug > Jan. The chemical score for histidine and threonine was more than the FAO recommended values. Overall, the nutritional quality of this unconventional species proved very good and comparable with other commercially important lean fishes.     

Fibrinolysis in Platelet Thrombi

The extent and duration of occlusive thrombus formation following an arterial atherothrombotic plaque disruption may be determined by the effectiveness of endogenous fibrinolysis. The determinants of endogenous fibrinolysis are the subject of much research, and it is now broadly accepted that clot composition as well as the environment in which the thrombus was formed play a significant role. Thrombi with a high platelet content demonstrate significant resistance to fibrinolysis, and this may be attributable to an augmented ability for thrombin generation and the release of fibrinolysis inhibitors, resulting in a fibrin-dense, stable thrombus. Additional platelet activators may augment thrombin generation further, and in the case of coronary stenosis, high shear has been shown to strengthen the attachment of the thrombus to the vessel wall. Neutrophil extracellular traps contribute to fibrinolysis resistance. Additionally, platelet-mediated clot retraction, release of Factor XIII and resultant crosslinking with fibrinolysis inhibitors impart structural stability to the thrombus against dislodgment by flow. Further work is needed in this rapidly evolving field, and efforts to mimic the pathophysiological environment in vitro are essential to further elucidate the mechanism of fibrinolysis resistance and in providing models to assess the effects of pharmacotherapy.     

The University of Toronto Continuous Flow Diffusion Chamber (UT-CFDC): A Simple Design for Ice Nucleation Studies

A new instrument, the University of Toronto Continuous Flow Diffusion Chamber (UT-CFDC), has been designed to study ice nucleation at low temperatures. Based on previous continuous flow instruments, it is a parallel plate model that minimizes convective instabilities by operating horizontally with the warmer plate on top. A variable position sample injector can account for effects arising from gravitational settling of ice particles that form. The residence time in the chamber can vary between 2.6 to 25 s and ice particle formation is monitored with a two-channel optical particle counter. Observation of homogeneous freezing of 100 nm sulfuric acid aerosols was used to verify the accuracy of the calculated relative humidities (RHs) in the chamber to be ±4%, where we report onset RHs for 0.1% of the particles freezing in the temperature range of 218 to 243 K. We also show that the chamber accurately establishes conditions of water saturation by conducting water uptake studies onto sulfuric acid aerosol at 243 K. The two channel OPC allows for ice and water droplet formation to be distinguished under such conditions. The chamber is a simple, cheap, and small design that can be readily assembled for laboratory studies.     

Chemically fixed nurse cells for culturing murine or primate embryonic stem cells

In current and past practice, murine or primate embryonic stem (ES) cells are usually cultured on live nurse cells for growth that keeps the cells in an undifferentiated state. It is troublesome, however, to prepare nurse cells for each cell culture and it is difficult to completely remove the nurse cells when they are transferred. In this study, mouse and monkey ES cells were therefore grown on chemically fixed mouse embryonic fibroblast (MEF) or human amniotic epithelial (HAE) cells. MEF cells were fixed by incubation in a glutaraldehyde or formaldehyde solution. HAE cells were immortalized by transfection of hTERT and chemically fixed with the same reagents. When mouse ES cells were cultured on these chemically fixed cells, the mouse ES cells grew well and expressed alkaline phosphatase, SSEA-1, and Oct-3/4 as their markers, indicating their undifferentiated state. The monkey ES cells also grew well and expressed alkaline phosphatase, SSEA-4, and Oct-4 as their markers, indicating their undifferentiated state. Freeze-drying HAE or MEF cells did not change their ability to support the undifferentiated growth of ES cells. Additionally, the chemically fixed cells could be utilized repeatedly in the culture of ES cells. These results demonstrate that chemically fixed nurse cells are useful for the maintenance of ES cells in an undifferentiated state in culture.     

Construction of diploid zygotes by interallelic complementation of ade6 in Schizosaccharomyces japonicus

The construction of diploid cells eases genetic analysis in haploid genetic systems because diploid cells allow for the characterization of essential genes. Here, we report the construction of diploid cells using ade6 point mutants that suppress each other via interallelic complementation in the fission yeast Schizosaccharomyces japonicus var japonicus (Sz. japonicus). We constructed an ade6-domK mutant in addition to the previously described ade6-domE. Phenotypes of both mutants exhibited adenine auxotrophy and red colonies. The mutations complemented the phenotypes in a mutually dependent manner. Diploid zygotes, in which the two mutations were introduced simultaneously into the same cells, were isolated by selecting for adenine independence. Such diploid cells are apparently larger in size than haploid cells, yet have a similar nuclear/cytoplasmic ratio, and thus the nuclear size control that has been reported in Sz. pombe is also present in Sz. japonicus.     

Contents of phthalate in polyvinyl chloride toys

The presence of plasticizers in PVC toys obtained in October 1998 was investigated. Diisononyl phthalate (DINP), di-2-ethylhexyl phthalate (DEHP), di-n-butyl phthalate (DBP), dinonyl phthalate (DNP), diheptyl phthalate (DHP), and di-2-ethylhexyl adipate (DEHA) were detected. The phthalates were found in all of the 68 samples. The principal phthalate found in toys was DINP, which was present in 48 of 68 samples. The DINP content ranged from 15 mg/g to 580 mg/g, and mean content was 308 mg/g. The highest content was found in a pacifier toy. DEHP was present in 20 of 68 samples and the content ranged from 2.0 mg/g to 380 mg/g. The mean content was 162 mg/g. It was found in 60% of domestic toys.     

Efficient expression, purification and characterization of mouse salivary alpha-amylase secreted from methylotrophic yeast, Pichia pastoris

We constructed a secretion vector of mouse salivary alpha-amylase, pPAM, using the AOX1 promoter-terminator and the secretion signal of 128 kDa pGKL killer protein, for an alternative yeast, Pichia pastoris. Taking advantage of multicopy insertion of the expression cassette and optimized growth conditions, we succeeded in highly efficient extracellular production (approximately 240 microg/ml) of mouse alpha-amylase in the 10 ml scale by conventional flask culture: this efficiency was about 90-fold higher than that of Saccharomyces cerevisiae. Growth temperature of cells was critical for efficient production of alpha-amylase. P. pastoris transformants secreted both core-glycosylated and non-glycosylated alpha-amylase molecules with a glycosylated:non-glycosylated ratio of about 20:80. Both glycosylated and non-glycosylated alpha-amylases were purified separately to apparent homogeneity. The signal sequence was correctly processed in both species, and the molecular masses of glycosylated and non-glycosylated alpha-amylase were determined to be 58 600 and 56 300, respectively, by mass spectrometry. We further studied the outer chain glycosylation of engineered mouse alpha-amylase secreted by P. pastoris.     

QoE assessment of fairness in networked game with olfaction: influence of time it takes for smell to reach player

In this paper, we carry out QoE (Quality of Experience) assessment of fairness between players in a networked game with olfaction. In the assessment, we investigate the influence of the time it takes for a smell to reach a player on the fairness. As the networked game, we deal with a fruit harvesting game in which two players play in a 3D virtual space, and we change the time it takes for a smell to reach a player by generating constant delays at two terminals. As a result, for example, we illustrate that the fairness is hardly damaged when the constant delays are smaller than about 500 ms.