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.

Triploid and tetraploid Zhikong scallop, Chlamys farreri Jones et Preston, produced by inhibiting polar body I

Triploid scallops are valuable for aquaculture because of their enlarged adductor muscle, and tetraploids are important for the commercial production of triploids. We tested tetraploid induction in the zhikong scallop by inhibiting polar body I in newly fertilized eggs. The ploidy of resultant embryos was determined by chromosome counting at 2- to 4-cell stage and by flow cytometry thereafter. Embryos from the control groups were mostly diploids (79%), along with some aneuploids. Embryos from the treated groups were 13% diploids, 18% triploids, 26% tetraploids, 13% pentaploids, and 36% aneuploids. Tetraploids, pentaploids, and most aneuploids suffered heavy mortality during the first week and became undetectable among the larvae at day 14. Five tetraploids (2%) were found among a sample of 267 spat from one of the replicates, and none was detected at day 450. The adductor muscle of triploid scallops was 44% heavier (P < .01) than that of diploids, confirming the value of the triploid technology in this species.

Effects of body size and water temperature on food consumption and growth in the sea cucumber Apostichopus japonicus (Selenka) with special reference to aestivation

To investigate the effects of body size and water temperature on feeding and growth in the sea cucumber Apostichopus japonicus (Selenka), the maximum rate of food consumption in terms of energy (C-maxe; J day(-1)) and the specific growth rate in terms of energy (SGRe; % day(-1)) in animals of three body sizes (mean +/- SE) - large (134.0 +/- 3.5 g), medium (73.6 +/- 2.2 g) and small (36.5 +/- 1.2 g) - were determined at water temperatures of 10, 15, 20, 25 and 30 degrees C. Maximum rate of food consumption in terms of energy increased and SGRe decreased with increasing body weight at 10, 15 and 20 degrees C. This trend, however, was not apparent at 25 and 30 degrees C, which could be influenced by aestivation. High water temperatures (above 20 degrees C) were disadvantageous to feeding and growth of this animal; SGRe of A. japonicus during aestivation was negative. The optimum temperatures for food consumption and for growth were similar and were between 14 and 15 degrees C, and body size seemed to have a slight effect on the optimal temperature for food consumption or growth. Because aestivation of A. japonicus was temperature dependent, the present paper also documented the threshold temperatures to aestivation as indicated by feeding cessation. Deduced from daily food consumption of individuals, the threshold temperature to aestivation for large and medium animals (73.3-139.3 g) was 24.5-25.5 degrees C, while that for small animals (28.9-40.7 g) was between 25.5 and 30.5 degrees C. These values are higher than previous reports; differences in sign of aestivation, experimental condition and dwelling district of test animals could be the reasons.

Seasonal variation of reproduction rates and body size of Calanus sinicus in the Southern Yellow Sea, China

Annual variations of egg production rate (EPR) and clutch size of Calanus sinicus, as well as body size of females (prosome length and dry weight), were investigated at a series of stations in the Southern Yellow Sea by onboard incubation. Calanus sinicus was spawning in all the 11 cruises investigated, and the annual variation of EPR was bimodal. Monthly average EPR was highest from May to July, respectively, 5.97, 5.36 and 6.30 eggs female(-1) d(-1), then decreased dramatically to only 1.37 eggs female(-1) d(-1) in August and attained the lowest 1.07 eggs female(-1) d(-1) in October. In November, average EPR increased again to 4.31 eggs female(-1) d(-1). Seasonal variation of clutch size was similar to EPR, except that it decreased gradually after August rather than dramatically as did EPR. Prosome length of females was maximum in May and minimum in October, but dry weight was highest in November. Monthly average EPR correlated better with prosome length than dry weight, while clutch size was rather determined by dry weight of females. It is suggested that egg production of C. sinicus was active during two discontinuous periods when both surface and bottom temperature fell into its favorite range (i.e. 10-23degreesC), and different reproductive strategies were adopted in these two reproductive peaks: other than the highest EPR, longer prosome length was also achieved by C. sinicus from May to July, while females in November developed shorter bodies but accumulated more energy for reproduction.

Metabolism of polychaete Neanthes japonica Izuka: relations to temperature, salinity and body weight

Polychaete Neanthes japonica is a species geographically specific in China and Japan with important scientific implication and commercial value. In this study, the relations of body weight, salinity and temperature to oxygen consumption and ammonia excretion of N. japonica were determined. Three different groups in body weight (large: 2.34 +/- 0.36 g, middle: 1.50 +/- 0.21 g and small: 0.62 +/- 0.12 g) were set for all experiments. Results show that the body weight is negatively related to the rates of oxygen consumption and ammonia excretion; and the relationship is significant. The oxygen consumption and ammonia excretion at 24A degrees C decreased at salinity from 5 to 30 and increased above 30, indicating that both lower and higher salinity are adverse and certain degree of salinity stress is necessary for enhancing the energy demand. At salinity 30, rising temperature from 18A degrees C to 30A degrees C, the oxygen consumption increased before 27A degrees C and then decreased. However, the relation of ammonia excretion and temperature seems more complex. Two-way ANOVA shows that salinity, temperature and body weight all have a significant effect on the oxygen consumption and ammonia excretion of the worm. Moreover, interaction between salinity/temperature and body weight is also significant. O:N (oxygen/nitrogen) ratio varies greatly in this case from 5.97 to 463.22, indicating that N. japonica can regulate the type of metabolic substrate against environment changes.

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.

Effects of diet and body size on phosphorus utilization of Liza haematocheila T. & S.

A 21-d laboratory experiment was conducted to study, the phosphorus (P) utilization of two different diets by redlip mullet Liza haematocheila T. & S. Sand-filtered water in salinity 30 and temperature 25 degrees C was used. Twenty-nine fish individuals were divided into three groups: 11 to group 1 (G1) fed on diet 1, 11 to group 2 (G2) fed on diet 2, and 7 to contrast group. Diet 1 was a commercial feed, more valuable in nutrition than diet 2 that similar to natural detritus. The results show the intake phosphorus (IP) of G1 was significantly higher than that of G2, and both increased linearly with body size at a certain amount of diet. The retention phosphorus (RP) in fish of G1 was lower than G2. The relationship between retention phosphorus and body size was positive and stronger in G2. Significant difference in faecal phosphorus (FP) was found between G1 and G2. Body size significantly impacted the excretion phosphorus (EP) in G1 but G2. The loss of intake phosphor-us in G1 was 10.83-20.27 mg per g fish weight gain, higher than that in G2 for 6.63-9.56. Of the phosphor-us, about 10% was allocated into growth, 50% in faeces, and the rest lost in excretion. The main part of phosphorus was lost in faeces but excretion. The phosphorus budget of the fish could be described as 100IP = 7.40RP + 47.39FP + 36.63EP (Diet 1) or 100IP = 11.93RP + 56.64FP + 21.76EP (Diet 2).

Effects of different light sources and illumination methods on growth and body color of shrimp Litopenaeus vannamei

Shrimps Litopenaeus vannamei with initial body weight of 2.108 +/- 0.036 g were sampled for specific growth rates (SGR) and body color measurements for 50 days under different light sources (incandescent lamp, IL; cool-white fluorescent lamp, FL; metal halide lamp, MHL; and control without lamp) and different illumination methods (illumination only in day, IOD, and illumination day and night, IDN). Body color of L. vannamei was measured according to the free astaxanthin concentration (FAC) of shrimp. The SGR, food intake (FI), feed conversion efficiency (FCE) and FAC of shrimps showed significant differences among the experimental treatment groups (P < 0.05). Maximum and minimum SGR occurred under IOD by MHL and IDN by FL, respectively (difference 56.34%). The FI of shrimp for the control group did not rank lowest among treatments, confirming that shrimp primarily use scent, not vision, to search for food. FI and FCE of shrimps were both the lowest among treatment groups under IDN by FL and growth was slow, thus FL is not a preferred light source for shrimp culture. Under IOD by MHL, shrimps had the highest FCE and the third highest FI among treatment groups ensuring rapid growth. FAC of shrimp were about 3.31 +/- 0.20 mg/kg. When under IOD by MHL and IDN by FL, FAC was significantly higher than the other treatments (P < 0.05). To summarize, when illuminated by MHL, L. vannamei had not only vivid body color due to high astaxanthin concentration but also rapid growth. Therefore, MHL is an appropriate indoor light source for shrimp super-intensive culture. SGR of shrimp was in significantly negative correlation to FAC of shrimp (P < 0.05). Thus, when FAC increased, SGR did not always follow, suggesting that the purpose of astaxanthin accumulation was not for growth promotion but for protection against intense light. (c) 2005 Elsevier B.V. All rights reserved.

Chromosome segregation in fertilized eggs from triploid Pacific oysters, Crassostrea gigas (Thunberg), following inhibition of polar body 1

Chromosome segregation in fertilized eggs from triploid Pacific oysters, following inhibition of the first polar body (PB1), was studied with acetic orcein staining techniques. To block the release of PB1, fertilized eggs were treated with 0.5 mg/l of cytochalasin B (CB). Four types of segregation were observed, namely, ''tripolar segregation'' (54.5%), ''united bipolar segregation'' (12%), ''separated bipolar segregation'' (2.5%), and ''incomplete united bipolar segregation'' (4%). The remaining 23% could not be classified because of chromosome disorganization, but appeared to be variants of the above. It seemed clear that the predominant pattern that gave rise to tetraploids was united bipolar segregation, although certain separated bipolar segregations might also lead to the formation of tetraploids. The sequential events of meioses observed in CB-treated eggs are described. The asynchrony of meiotic events and possible mechanisms for the various types of chromosome segregation are discussed.

Photoperiodic regulation in energy intake, thermogenesis and body mass in root voles (Microtus oeconomus)

The present study was designed to examine whether photoperiod alone was effective to induce seasonal regulations in physiology in root voles (Microtus oeconomus) from the Qinghai-Tibetan plateau noted for its extreme cold environment. Root voles were randomly assigned into either long photoperiod (LD; 16L: 8D) or short photoperiod (SD; 8L: 16D) for 4 weeks at constant temperature (20 degrees C). At the end of acclimation, SD voles showed lower body mass and body fat coupled with higher energy intake than LD voles. SD greatly enhanced thermogenic capacities in root voles, as indicated by elevated basal metabolic rate (BMR), nonshivering thermogenesis (NST), mitochondrial protein content and uncoupling protein-1 (UCP1) content in brown adipose tissue (BAT). Although no variations in serum leptin levels were found between SD and LD voles, serum leptin levels were positively correlated with body mass and body fat mass, and negatively correlated with energy intake and UCP1 content in BAT, respectively. To summarize, SD alone is effective in inducing higher thermogenic capacities and energy intake coupled with lower body mass and body fat mass in root voles. Leptin is potentially involved in the photoperiod induced body mass regulation and thermogenesis in root voles. (c) 2006 Elsevier Inc. All rights reserved.

Seasonal thermogenesis and body mass regulation in plateau pikas (Ochotona curzoniae)

Changes in photoperiod, ambient temperature and food availability trigger seasonal acclimatization in physiology and behavior of many animals. In the present study, seasonal adjustments in body mass and in several physiological, hormonal, and biochemical markers were examined in wild-captured plateau pikas (Ochotona curzoniae) from the Qinghai-Tibetan plateau. Our results showed that plateau pikas maintained a relatively constant body mass throughout the year and showed no seasonal changes in body fat mass and circulating levels of serum leptin. However, nonshivering thermogenesis, cytochrome c oxidase activity, and mitochondrial uncoupling protein 1 (UCP1) contents in brown adipose tissues were significantly enhanced in winter. Further, serum leptin levels were positively correlated with body mass and body fat mass while negatively correlated with UCP1 contents. Together, these data suggest that plateau pikas mainly depend on increasing thermogenic capacities, rather than decreasing body mass, to cope with cold, and leptin may play a potential role in their thermogenesis and body mass regulation.

Seasonal changes in weight and body composition of yak grazing on alpine-meadow grassland in the Qinghai-Tibetan plateau of China

Forty-five male yaks (born April 2001) were studied to determine how seasonal changes on the Qinghai-Tibetan plateau affected BW and body composition. Thirty yaks were weighed monthly from birth to 26 mo of age to determine seasonal changes in BW. The remaining 15 yaks were allocated randomly to five groups (three yaks per group), designated for slaughter at 13, 15, 18, 22, and 25 mo to measure seasonal effects on body chemical composition. All yaks were grazed on the alpine-meadow grassland of the plateau without any supplementation. All BW and body composition data were calculated on an individual basis. Body weight and body composition data were both compared across seven growth periods spanning 2 yr and defined by season. From April (birth) to December 2001 of the first growing season, yak BW increased (P < 0.01); however, during the subsequent cold season (December 2001 to May 2002), BW decreased (P < 0.01). The second growing season ran from May 2002 (13 mo of age) to October 2002 (18 mo of age), and the second live weight-loss season ran from October 2002 until May 2003. The weight loss experienced by yaks during the first weight loss season was 25.64% of the total weight gain in the first growing season. The weight loss experienced by yaks during the second weight-loss season was 29.73% of the total weight gain in the second growing season. Energy retention in the second growing season was 291.07 MJ, 50.8% of which was consumed during the subsequent cold season. Energy accumulation in the summer (from May to July) and fall (from July to October) of the second growing season did not differ (5.01 and 6.30 MJ/kg of EBW gain, respectively; P = 0.63). The energy mobilized during the second winter (from October 2002 to February 2003) was 16.49 MJ/kg of EBW, and in the second spring (from February to May 2003), it was 9.06 MJ/kg of EBW. These data suggest that the decrease in grazing yak BW during the first cold season is much less than during the second cold season, and that the energy content per unit of BW mobilized is greater (P = 0.02) in winter than in spring. Results from this study demonstrate highly efficient compensatory growth in grazing yaks following the first weight loss period during the first cold season. This benefit could be exploited by herders to improve yak production. Yaks may have developed a type of self-protection mechanism to overcome the long cold seasons in the Qinghai-Tibetan plateau.

柴西南区下干柴沟组下段层序地层格架内沉积相展布及有利储集砂体分布研究

Lower member of the lower Ganchaigou Formation in the southwestern of Qaidam Basin is one of the main targeted exploration zones. With the advancement of exploration, the targets are gradually switching into the lithologic reservoirs and it is urgent to gain the more precise research results in distribution of sedimentary facies and sandstones. Guided by the theory of sequence stratigraphy and sedimentology as well as on the basis of many logging data, drillings, seismic data and chemical tests, the paper comprehensively analyzes the sedimentary facies and sandstones in the lower member of lower Ganchaigou Formation in the southern of Chaixi. According to the identification marks of the key interface in sequence stratigraphy, the key interfaces in lower member of lower Ganchaigou Formation in the southwestern of Qaidam Basin are identified as two third-order sequences SQ1、SQ2. By calibrating the synthetic seismogram, the seismic sequence, well drilling and logging sequences are united. Based on the works above, this paper chooses seven primary cross-sections and builds connecting-well stratigraphic correlation of seven main connecting-well sections. Ultimately, the high-resolution sequence stratigraphic frameworks in the lower member of the lower Ganchaigou Formation, which are uniform to logging and seismic data, are figured out. In terms of study on each sequence features, the main style of the base-level cycle overlay which forms the third-order sequence is confirmed. It contains asymmetric “becoming deep upward” style and symmetry style. Researching on the spreading characters of sequence stratigraphy indicates that SQ1 and SQ2 are rather thicker near northwest well Shashen 20 and Shaxin1 while they are quite thiner near Hongcan 1, Yuejin, Qie 4 and Dong8-Wu3, and the thickness of SQ1 is thicker than SQ2.Based on the deep analysis of the marks for depositional facies, it is proposed that the lake facies and braid river deltas facies mainly occurred in study areas. Besides, the sorts of sub-facies and micro-facies model are divided and described. Under the control of high-resolution sequence stratigraphic framework, three source directions from Arlarer Mountain、Qimantage Mountain and Dongchai Mountain are identified by using the features of heavy mineral assemblage and paleogeomorphy. In addition, regularities of distribution sedimentary facies in sequence stratigraphic framework are studied in accordance with research thinking of the "point" (single well) "line" (section) "face" (plane). In the stage of lower member in the lower Ganchaigou Formation in the southwestern of Qaidam Basin, it is at the early phrase of evolution of the lake basin with the gradual outspread and the rise of the lake level. Combined with physical analysis of reservoir sands formed in different sedimentary environment, the paper studies the style of favorable sandstone bodies that are underwater distributary channel of braided rive delta front, coarse sand in mouth bar and the sand body in sand flat of shore-shallow lacustrine facies. Finally, this article comprehensively analyzes the distribution relationship between sedimentary facies and favorable sandstone body and proposes the ideas that sequence SQ1 Yuejin area, well east 8-wu3 area, well qie4-qie1 area and well hongcan2 area are distributed areas of favorable sandstone.