Thermal stability of carp G-actin monitored by loss of polymerization activity using an extrinsic fluorescent probe

ABSTRACT:   The thermal stability of carp G-actin was investigated by monitoring loss of actin polymerization ability. To determine the amount of native actin remaining after heat treatment, actin was labeled with a fluorescence reagent, N-(1-pyrene)iodoacetamide. The loss of polymerization ability of carp actin during heat treatment, at between 45 and 55°C, occurred faster than that of chicken actin. The inactivation rate was influenced by concentrations of ATP and Ca²⁺ in solution. With the increase of Ca²⁺ concentration, the inactivation of carp actin was markedly suppressed. Furthermore, the activation energy of the inactivation of carp actin obtained from an Arrhenius plot was similar to that of chicken actin. These results indicated that the thermal instability of carp G-actin was due to the low affinites of ATP and Ca²⁺ for carp actin described in a previous report.     

Bioaccumulation of cadmium and its biochemical effect on selected tissues of the catfish (<Emphasis Type="Italic">Clarias gariepinus</Emphasis>)

The present study examines the pattern of accumulation of cadmium (Cd) and its biochemical effects on selected tissues of a variety of African catfish (Clarias gariepinus), after exposure to various doses of Cd. The results obtained indicate that at the end of 21 days of exposure, the total tissue organ cadmium concentration followed the pattern kidney > gill > liver > muscle for each of the exposure concentrations. The levels of Cd in these organs were higher than those in ambient water. Moreover, while the rate of uptake of Cd increased with time in the kidney, liver and muscle, it decreased in the gill. Superoxide dismutase (SOD) was significantly elevated only in the kidney of catfish treated with 0.2 and 0.4 ppm of Cd for 7 days compared with the control. Conversely, gill SOD was significantly decreased in the same concentrations of Cd-treated catfish relative to the control. Statistically similar levels of SOD were observed in the liver, brain and muscle with all the treatments after the same duration of treatment. In the fish exposed for 21 days, SOD activity was significantly decreased in the kidney with a corresponding increase in lipid peroxidation (LPO), but it manifested only with the 0.2- and 0.4-ppm Cd treatment relative to the control. In the liver, however, Cd exposure significantly increased SOD in the 0.2- and 0.4-ppm treatments of the same duration. In conclusion, the present study indicates that the accumulation of Cd and its effect on SOD and LPO in C. gariepinus is dependent on concentration, tissue and time     

Differential HSP90α expression in fish hepatocytes from polluted estuary during summer

The molecular chaperone, heat shock protein 90α (HSP90α), plays an important role in protein folding, degradation of denatured proteins and steroid activation. It is essential for the maintenance of cellular integrity and survival when induced in response to environmental, physical and chemical stresses. In the present investigation the effect of environmental stress on HSP90α expression was examined in grey mullet Mugil cephalus living in either a contaminated (Ennore) or uncontaminated (Kovalam) estuary over two seasons: Hepatocytes were isolated from grey mullet of both estuaries. Oxidative stress was determined along with HSP90α in these fish. Additionally, immunohistochemical changes were studied to confirm the HSP90α expression. Comparison of the results revealed enhanced hepatocyte oxidative stress and HSP90α expressions in fish from Ennore to a significant extent than fish from Kovalam. Also, the results showed significant seasonal variations with maximum expression observed during summer compared to the monsoon season. Overexpression of HSP90α in hepatocytes exposed to chronic environmental stress by pollutants may confer differential effects on cell survival by protecting against oxidative stress induced changes. The results also indicate that seasonal variations have significant effect on the HSP90α expression.     

Specific dynamic action in two body size groups of the southern catfish (<Emphasis Type="Italic">Silurus meridionalis</Emphasis>) fed diets differing in carbohydrate and lipid contents

Specific dynamic action (SDA), the energy costs associated with meal digestion and assimilation, is generally affected by body size and food composition. We assessed the postprandial metabolic response and calculated SDA in two size groups of the southern catfish (Silurus meridionalis), each fed one of two diets, high lipid or high carbohydrate, at a meal size of 4% the body mass. Using a continuous-flow respirometer, we determined the oxygen consumption rate at 2-h intervals until the postprandial oxygen consumption rate returned to the prefeeding level. None of the parameters (resting metabolic rate, Rpeak, factorial ratio, time-to-peak, duration, energy expended on SDA, or SDA coefficient) were significantly affected by diet nor was there an interaction between diet and body mass. Rpeak and energy expended on SDA for the whole fish body were significantly higher in the larger fish than the smaller one in both dietary treatments, whereas no significant effect of body size was found in mass specific values. Factorial ratio (range 3.41 to 3.60), peak time (range 9.6 to 12.7 h), SDA coefficient (range 9.36 to 10.36%), and SDA duration (range 62.0 to 71.0 h) did not significantly differ between body size groups. These results suggest that in S. meridionalis the percentage of assimilated energy allocated to SDA may be independent of the body mass.     

Evaluation of three phytoplankton species as food for the pearl oyster <Emphasis Type="Italic">Pinctada fucata</Emphasis>

In the pearl cultivation farms of the Ehime Prefecture, Japan, mass mortalities of the pearl oyster Pinctada fucata have occurred since 1994. The occurrences of mass mortality roughly coincided with a shift of the dominant phytoplankton from Skeletonema and Chaetoceros to Chaetoceros and Nitzschia all of which belong to Bacillariophyceae. Hence, we evaluated Nitzschia, together with Chaetoceros and Isocrysis, as food for the oyster. Wet weights, lengths, widths, glycogen contents, and growth rates in terms of wet weight of the oysters in all the feeding treatments were significantly higher than those in the non-feeding treatment. The highest glycogen content (2.34%) and growth rate (2.21 g month⁻¹) were found in the Chaetoceros treatment. Growth rate in the Isocrysis treatment (1.63 g month⁻¹) was also high, although glycogen content in this treatment (0.41%) was low. In the Nitzschia treatment, growth rate of the oyster (0.94 g month⁻¹) was the lowest and glycogen content (0.83%) was also low relative to that in the Chaetoceros treatment. Chlorophyll a concentration in fecal pellets was lowest in the Nitzschia treatment (<2.7 μg mg⁻¹), suggesting more complete digestion of Nitzschia by the oyster. Thus, Nitzschia was edible and digestible but not assimilated by P. fucata. We propose the following scenario for the relationship between Nitzschia dominance and mass mortality. When Nitzschia dominates in a culture area, the physiological condition of P. fucata deteriorates due to low assimilation of Nitzschia by the oyster, followed by susceptibility of the oyster to infection by agents lethal to the oyster.     

The harvest of the freshwater crayfish <Emphasis Type="Italic">Astacus leptodactylus</Emphasis> Eschscholtz in Turkey: harvest history,impact of crayfish plague,and present distribution of harvested populations

This review focuses on the present distribution of populations of the crayfish Astacus leptodactylus that are harvested in Turkey. It also examines the history of this harvest and the impact that crayfish plague has had on them. Crayfish plague, caused by the fungus-like organism, Aphanomyces astaci Schikora, 1906, is a severe parasite of freshwater crayfish and has caused a lot of damage to A. leptodactylus populations in Turkey since 1984. Turkey was the largest provider of A. leptodactylus to Western Europe from 1970 (or possibly earlier) until 1986. For example, the peak production was reached in the early 1980s, with over 5,000 tonnes being exported in 1984. On the other hand, as a result of the crayfish plague the harvest of A. leptodactylus was reduced severely in most populations in Turkey after 1985. The harvest was only 320 tonnes in 1991. After the occurrence of crayfish plague in Turkey, in order to increase crayfish production uncontrolled A. leptodactylus stockings have been carried out in many waterbodies throughout Turkey. These introductions have caused an increase in the number of A. leptodactylus populations, but exploitation of A. leptodactylus is still under the pressure of the plague, although there has been a steady increase in crayfish production in recent years. The harvest increased to 2,317 tonnes in 2004. Fortunately, among those populations affected by crayfish plague, large amounts of A. leptodactylus can still be harvested from three lakes, ?znik (Bursa), E?irdir (Isparta) and Çivril (Denizli). Thus, it seems that A. leptodactylus has a degree of resistance to crayfish plague. It is therefore interesting to investigate the resistance of A. leptodactylus caught from these populations to crayfish plague.     

Transpositional feeding rhythm of loach <Emphasis Type="Italic">Misgurnus anguillicaudatus</Emphasis> from larvae to juveniles and its ontogenesis under artificial rearing conditions

The diel feeding rhythm and ontogenesis during early life stage of loach Misgurnus anguillicaudatus were investigated under experimental conditions (light: L 06:00–18:00, D 18:00–06:00 h). Morphological and behavioral developments of loach from newly hatched to 40 days after hatching were observed. Larvae were able to prey on daphnia 3–4 days after hatching at 23 ± 0.5°C. As the larvae grew, they showed an increasing feeding capacity and a distinct feeding rhythm. Feeding intensity and incidence for day-4 larvae were highest at 10:00 and 16:00 h. The highest levels of feeding intensity for day-12 larvae occurred at 08:00, 12:00, and 18:00 h as did feeding incidence. By day 20, when the larvae metamorphosed, the highest levels of feeding intensity occurred at 06:00, 18:00, and 24:00 h and were concurrent with the highest feeding incidence. After metamorphosis, feeding capacity had again increased considerably and, in contrast to the earlier stages before day 20, feeding intensity for day-30 juveniles peaked at 05:00 and 20:00 h, about 1–2 h after the maximum feeding incidence. The feeding rhythm of loach juveniles at day 40 was almost the same as the day-30 juveniles. The estimated maximum daily feeding rates were 43.1%, 33.4%, 19.0%, 12.8%, and 5.8% of body weight on days 4, 12, 20, 30, and 40, respectively. Thus, loach was found to have different feeding rhythms in the pre- and post-metamorphosis stages, with the highest feeding activity in daytime during the larval planktonic stage before metamorphosis, and intensely nocturnal feeding behavior during the juvenile benthic stage after metamorphosis.     

育珠期三角帆蚌的生长及其与珍珠增长的关系

通过对三角帆蚌壳长、壳宽、蚌总重（整体湿重）、内脏团湿重、壳重和珍珠重等指标在5-11月间的逐月连续测定,研究了常规养殖模式下育珠期2龄三角帆蚌在主要生长季节的生长规律及其与珍珠生长的相关性。结果表明：珍珠重(PW)与壳长(SL)、壳宽(SW)的关系分别为PW=0.0008SL3.3946 (R2=0.6948)和PW=0.7809SW1.0227 (R2=0.6888);而珍珠重(PW)与蚌重(TW)、内脏团重(BW)和壳重(W)的关系分别为PW=0.0021TW1.5434 (R2=0.7337),PW=0.107BW0.9125 (R2=0.7158)和PW=0.0324W1.1259 (R2=0.7101);三角帆蚌总重(TW)与壳长(SL)、 壳宽(SW)的关系最适合用指数函数进行拟合,其关系式分别为TW=16.003e0.1681SL,(R2=0.7961),和TW=64.311e0.1372SW (R2=0.6822);而内脏团重(BW)和壳重(W)与壳长(SL)、壳宽(SW)的关系则适合为幂函数曲线拟合,关系式分别为BW=0.0188SL3.1427,(R2=0.6927);W=0.0656SL2.7721, (R2=0.8271)和BW=12.446SW0.8974 (R2=0.617);W=19.876SW0.802 (R2=0.7563)。本研究结果发现,珍珠生长与蚌总重、壳长和壳宽等外部测量指标之间的相关性显著,从而无须通过杀蚌取珠而直接通过这些外部生长指标的测定就能较好地了解珍珠的生长,更好地为珍珠养殖生产和管理提供指导。     

CYP450 epoxygenase/EET pathway attenuates adipose inflammation of obese mice through HIF-1α

AIM To investigate the effects of cytochrome P450 （CYP450） epoxygenase/epoxyeicosatrienoic acid （EET） pathway on insulin resistance in obese mice, and to explore the possible mechanisms. METHODS High-fat diet-induced obesity model was established in C57BL/6Cnc mice, and the obese mice were randomly divided into 3 groups, including obesity group （treated with saline; n=10）, EET group （treated with 11,12-EET; n=10） and EET inhibitor 14,15-epoxyeicosa-5（Z）-enoic acid （EEZE） group （n=10）. Normal C57BL/6Cnc mice （n=10） treated with saline served as control. Protein expression of CYP2J2 （one of CYP450 epoxygenases） and hypoxia-inducible factor-1α （HIF-1α） was measured by Western blot. Vessel-like structure was detected by immunofluorescence staining. The serum levels of insulin, tumor necrosis factor-α （TNF-α）, interleukin （IL）-1β, IL-6 and monocyte chemoattractant protein-1 （MCP-1） were measured by ELISA. RESULTS In obese mice, homeostasis model assessment of insulin resistance （HOMA-IR） values were increased, the protein level of CYP2J2 was reduced, and the protein level of HIF-1α was increased in adipose tissues as compared with the controls （P<0.05）. The serum levels of MCP-1, IL-1β, IL-6 and TNF-α were also significantly increased in obese mice （P<0.05）. After treatment with 11, 12-EET, the HOMA-IR values were decreased compared with vehicle-treated obese mice, HIF-1α expression levels were decreased in the adipose tissue, and the serum levels of MCP-1, IL-1β, IL-6 and TNF-α were reduced （P<0.05）. Immunohistochemical results of adipose tissue from vehicle-treated obese mice showed a marked decrease in vessel-like structures （CD31-positive） compared with normal control mice （P<0.05）. EET treatment significantly increased the newly formed vessel-like structures in the visceral adipose tissues of obese mice as compared with vehicle-treated obese mice （P<0.05）. CONCLUSION High-fat diet-induced obesity and insulin resistance are closely related to the CYP450 pathway. Exogenous EETs effectively decrease obesity-induced insulin resistance possibly through pro-angiogenesis and attenuation of hypoxia and inflammation.     

陆地棉脱水素基因GhDHN1启动子序列分析

为了筛选棉花抗逆转基因育种的候选启动子,根据陆地棉脱水素基因GhDHN1的cDNA序列和陆地棉基因组序列,利用生物信息学分析方法,获得棉花陆地棉脱水素基因GhDHN1启动子序列。结果表明,该启动子上多个位点含有启动子的基本元件TATA-box和CAAT-box,并含有非生物逆境胁迫响应元件、响应植物激素顺式作用元件、多种光调控相关的顺式作用元件以及胚乳表达顺式调控元件等。推测GhDHN1基因的启动子受光诱导,同时受低温和干旱胁迫等非生物逆境的诱导,参与棉花的生长发育。