Evaluation of different culture conditions for high-level soluble expression of human cyclin A(2) with pET vector in BL21 (DE3) and spectroscopic characterization of its inclusion body structure

In this paper, we evaluated various parameters of culture condition affecting high-level soluble expression of human cyclin A, in Escherichia coli BL21(DE3), and demonstrated that the highest protein yield was obtained using TB(no glycerol) + 0.5% glucose medium at 25 degrees C. By single immobilized metal ion affinity chromatography, we got highly purified human cyclin A(2) with a yield ranged from 20 to 30 mg/L. By amyloid-diagnostic dye ThT binding and Fourier transform infrared spectroscopy, we observed a significant decrease in alpha-helix content and an increase in beta-sheet structure in cyclin A(2) inclusion body in comparison to its native protein, and confirmed the resemblance of the internal organization of cyclin A(2) inclusion body and amyloid fibrils.

Domain decomposition hybrid method for numerical simulation of bluff body flows:theoretical model and application

The discrete vortex method is not capable of precisely predicting the bluff body flow separation and the fine structure of flow field in the vicinity of the body surface. In order to make a theoretical improvement over the method and to reduce the difficulty in finite-difference solution of N-S equations at high Reynolds number, in the present paper, we suggest a new numerical simulation model and a theoretical method for domain decomposition hybrid combination of finite-difference method and vortex method. Specifically, the full flow. field is decomposed into two domains. In the region of O(R) near the body surface (R is the characteristic dimension of body), we use the finite-difference method to solve the N-S equations and in the exterior domain, we take the Lagrange-Euler vortex method. The connection and coupling conditions for flow in the two domains are established. The specific numerical scheme of this theoretical model is given. As a preliminary application, some numerical simulations for flows at Re=100 and Re-1000 about a circular cylinder are made, and compared with the finite-difference solution of N-S equations for full flow field and experimental results, and the stability of the solution against the change of the interface between the two domains is examined. The results show that the method of the present paper has the advantage of finite-difference solution for N-S equations in precisely predicting the fine structure of flow field, as well as the advantage of vortex method in efficiently computing the global characteristics of the separated flow. It saves computer time and reduces the amount of computation, as compared with pure N-S equation solution. The present method can be used for numerical simulation of bluff body flow at high Reynolds number and would exhibit even greater merit in that case.

Evaluation of induced triploid shrimp Penaeus (Fenneropenaeus) chinensis cultured under laboratory conditions

Triploid Penaeus (Fenneropenaeus) chinensis was successfully produced by heat shock. Their metamorphosis, the relationship between body weight and length and difference in appearance between triploids and their diploid siblings under laboratory culture were studied. Hematological studies showed a smaller number of haemocytes, but larger cell volume, in triploids than in diploids. Triploid shrimp did not show higher growth during the immature stage, but exhibited superior growth during the maturation stage. Characteristics of reproductive organs indicated that triploid shrimp may be sterile and sex ratio can be changed through triploidization of shrimp. This paper summarizes the progress made in triploid shrimp research which would be helpful in understanding more about triploids of crustaceans. (c) 2006 Elsevier B.V. All rights reserved.

C, N and P regeneration by a detritivorous fish, Liza haematocheila T. & S.: effects of temperature, diet and body size

Detritus, as a nutrients reservoir, affects the trophic structure and dynamics of communities and supports a greater diversity of species and longer food chains. Detritivorous fish is an important organism to regenerate the nutrients from sediments. Despite the numerous studies on the nutrients cycle in fish, only a few attempts have been made to quantify the regenerating ability. In the present study, we chose the common detritivorous fish redeye mullet as the research object. Redeye mullet is also a common poly-culture fish in China. Diet, including a commercial diet mostly used in aquaculture and a home-made diet with contents close to detritus, was used and considered as a fixed factor. Temperature was also considered as a fixed factor as much research has shown that temperature has significant effects on fish metabolism. Moreover, body size was regarded as a covariate under analysis of covariance. Three key nutrients, namely carbon, nitrogen and phosphorus, were used to measure the nutrient-regenerating ability of redeye mullet under laboratory conditions. The results showed that the nutrient regeneration in percent of the consumption decreased with increasing temperature. Carbon and nitrogen regeneration of redeye mullet fed on commercial diet was lower than those of the home-made diet group, while the opposite was found for phosphorus. In each group, the amount of regenerated nutrients increased linearly with body size. Fed on the home-made diet, 5-g fish at 25 degrees C can regenerate 210.822 mg C, 37.533 mg N and 0.727 mg P per day.