Effects of adrenergic agonists on the extrahepatic expression of vitellogenin Ao1 in heart and brain of the Chinese rare minnow (Gobiocypris rarus)

Teleost vitellogenins (VTGs) are large multidomain apolipoproteins and traditionally considered as the estrogen responsive precursors of the major egg yolk proteins. We identified five clones encoding VTGs, about 16% of the random EST clones from our constructed cDNA library from Chinese rare minnow liver tissue treated with 17 beta-estradiol (E2). Full-length vtgAo1 has been obtained based on the sequence information of four partial cDNA inserts by RACE. The inducibility of the vtgAo1 expression in liver by E2 was confirmed by RT-PCR. The presence of vtgAo1 transcripts have been observed primarily in liver. However. a significant level of the vtgAo1 was found in an unexpected location, heart, particularly in atrial cells by RT-PCR and whole mount in situ hybridization analyses. The vtgAo1 mRNA expression in heart and liver tissue could be suppressed by both alpha-adrenergic agonist, phenylephrine (PE) and beta-adrenergic agonist, isoproterenol (ISO). The expression of VTG in the heart observed in the present studies suggested it may provide protection from surplus intracellular lipids in fish cardiomyocytes as triglyceride transport proteins do in mammals. The results also indicated that the production of teleost vtg in vivo can be regulated by riot only estrogenic agents, but adrenergic signals as well. (c) 2008 Elsevier B.V. All rights reserved.

UV-B-induced Oxidative Damage and Protective Role of Exopolysaccharides in Desert Cyanobacterium Microcoleus vaginatus

UV-B-induced oxidative damage and the protective effect of exopolysaccharides (EPS) in Microcoleus vaginatus, a cyanobacterium isolated from desert crust, were investigated. After being irradiated with UV-B radiation, photosynthetic activity (Fv/Fm), cellular total carbohydrates, EPS and sucrose production of irradiated cells decreased, while reducing sugars, reactive oxygen species (ROS) generation, malondialdehyde (MDA) production and DNA strand breaks increased significantly. However, when pretreated with 100 mg/L exogenous EPS, EPS production in the culture medium of UV-B stressed cells decreased significantly; Fv/Fm, cellular total carbohydrates, reducing sugars and sucrose synthase (SS) activity of irradiated cells increased significantly, while ROS generation, MDA production and DNA strand breaks of irradiated cells decreased significantly. The results suggested that EPS exhibited a significant protective effect on DNA strand breaks and lipid peroxidation by effectively eliminating ROS induced by UV-B radiation in M. vaginatus.

Growth and antioxidant system of Escherichia coli in response to microcystin-RR

Microcystins are a kind of cyclic hepatoxins produced by many species of cyanobacteria. The toxic effects of microcystins on animals and plants have been well studied. However, the reports about the effects of microcystins on microbial cells are very limited. In present paper, Escherichia coli was undertaken to determine the effect of microcystin-RR. These results suggested that microcystin-RR could prolong the growth of E. coli when exposed to high concentrations of microcystin-RR and cause the accumulation of ROS and induce the oxidant stress for a short time. The antioxidant system protects E. coli from oxidative damage.

Accumulation of hepatotoxic microcystins in freshwater mussels, aquatic insect larvae and oligochaetes in a large, shallow eutrophic lake (Lake Chaohu) of subtropical China

This paper studied the seasonal changes of two common microcystins (MCs), MC-RR and -LR, in the commercially important mussel Corbicula fluminea in Lake Chaohu, where there occurred dense cyanobacteria. Occasional measurements were also made for MC in the mussel Arconaia lanceolat, the oligochaete Limnodilus hoffineisteri and the insect larva Chironomus sp. Mean MC of C. fluminea was much higher in hepatopancreas than in intestine and foot. Our study is the first to report accumulation of MCs in oligochaetes and aquatic insect larvae. The hi-h contents of MCs in the insect larvae suggest a great possibility for the transfer of MCs to benthos-feeding omnivores like common carp. According to the provisional standard by the WHO, 28.6% of the collected C. fluminea were harmful for human consumption, assuming a daily consumption of 300 by a person. It is recommended that edible mussels should not be collected for human consumption during toxic cyanobacterial blooms in Lake Chaohu.

Inhibition of no tail (ntl) gene expression in zebrafish by external guide sequence (EGS) technique

External guide sequence (EGS) technique, a branch of ribozyme strategy, can be enticed to cleave the target mRNA by forming a tRNA-like structure. In the present study, no tail gene (ntl), a key gene participating in the formation of normal tail, was used as a target for ribonuclease (RNase) P-mediated gene disruption in zebrafish in vivo. Transient expression of pH1-m3/4 ntl-EGS or pH1-3/4 ntl-EGS produced the full no tail phenotype at long-pec stage in proportion as 24 or 35%, respectively. As is expected that the full-length ntl mRNA of embryos at 50% epiboly stage decreased relative to control when injected the embryos with 3/4 EGS or m3/4 EGS RNA. Interestingly, ntl RNA transcripts, including the cleaved by EGS and the untouched, increased. Taken together, these results indicate that EGS strategy can work in zebrafish in vivo and becomes a potential tool for degradation of targeted mRNAs.

Cloning, expression and characterization of aerolysin from Aeromonas hydrophila in Escherichia coli

Aerolysin is a toxin (protein in nature) secreted by the strains of Aeromonas spp. and plays all important role in the virulence of Aeromonas strains. It has also found several applications such as for detection of glycosylphosphatidylinositol (GPI)-anchored proteins etc. A. hydrophila is a ubiquitous Gram-negative bacterium which causes frequent harm to the aquaculture. To obtain a significant amount of recombinant aerolysin in the active form, in this study, we expressed the aerolysin in E. Coli Under the control of T7 RNase promoter. The coding region (AerA-W) of the aerA gene of A. hydrophila XS91-4-1. excluding partial coding region of the signal peptide was cloned into the vector pET32a and then transformed into E. coli b121. After optimizing the expression conditions, the recombinant protein AerA-W was expressed in a soluble form and purified using His-Bind resin affinity chromatography. Recombinant aerolysin showed hemolytic activity in the agar diffusive hemolysis test. Western blot analysis demonstrated good antigenicity of the recombinant protein.

Studies on the restoration succession of PFU microbial communities in a pilot-scale microcosm

In order to imitate the restoration succession process of natural water ecosystem, a laboratory microcosm system of constant-flow-restoration was designed and established. A eutrophycation lake, Lake Donghu, was selected as the subject investigated. Six sampling stations were set on the lake, among which the water of station IV was natural clean water, and others were polluted with different degrees. Polyurethane foam unit microbial communities, which had colonized in the stations for a month, were collected from these stations and placed in their respective microcosms, using clean water of station IV to gradually replace the water of these microcosms. In this process, the healthy community in clean water continuously replaced the damaged communities in polluted water, the restoration succession of the damaged communities was characterized by weekly determination of several functional and structural community parameters, including species number (S), diversity index (DI), community pollution value (CPV), heterotrophy index (HI), and similarity coefficient. Cluster analysis based on similarity coefficient was used to compare the succession discrepancies of these microbial communities from different stations. The ecological succession of microbial communities during restoration was investigated by the variable patterns of these parameters, and based on which, the restoration standards of these polluted stations were suggested in an ecological sense. That was, while being restored, the water of station 0 (supereutrophycation) should be substituted with natural clean water by 95%; station I (eutrophycation), more than 90%; station II (eutrophycation), more than 85%; station III (eutrophycation), about 85%; station V (mesoetitrophycation), less than 50%. The effects of the structural and functional parameters in monitoring and assessing ecological restoration are analyzed and compared. (C) 2007 Elsevier Ltd. All rights reserved.

Development of the microbial communities in Lake Donghu in relation to water quality

There is increasing recognition that protozoa is very useful in monitoring and evaluating water ecological healthy and quality. In order to study the relationship between structure and function of protozoan communities and water qualities, six sampling stations were set on Lake Donghu, a hypereutrophic subtropical Chinese lake. Microbial communities and protists sampling from the six stations was conducted by PFU (Polyurethane foam unit) method. Species number (S), diversity index (DI), percentage of phytomastigophra, community pollution value (CPV), community similarity and heterophy index (HI) were mensurated. The measured indicators of water quality included total phosphorus (TP), dissolved oxygen (DO), Chemical oxygen demand (COD), NH4 (+), NO2 (-) and NO3-. Every month water samples from stations I, II, III, IV were chemically analyzed for a whole year, Among the chemically analyzed stations, station I was the most heavily polluted, station II was the next, stations III and IV had similar pollution degrees. The variable tendencies of COD, TP, NH3, NO2-, NO3-, and DO during the year was approximately coincident among the six stations. Analysis from the community parameters showed that the pollution of station 0 was much more serious than others, and station V was the most slight. Of the community parameters, CPV and HI were sensitive in reflecting the variables of the water quality. Community similarity index was also sensitive in dividing water qualities and the water quality status of different stations could be correctly classified by the cluster analysis. DI could reflect the tendency of water quality gradient, species number and percentage of Phytomastigophora was not obvious in indicating the water quality gradient.

Use of microbial community to evaluate performance of a wetland system in treating Pb/Zn mine drainage

The performance of a wetland system in treating lead (Pb)/zinc (Zn) mine drainage was evaluated by using the polyurethane foam unit (PFU) microbial community (method), which has been adopted by China as a standardized procedure for monitoring water quality. The wetland system consisted of four cells with three dominant plants: Typha latifolia, Phragmites australis and Paspalum distichum. Physicochemical characteristics [pH, EC, content of total suspended solid (TSS) and metals (Pb, Zn, Cd, and Cu)] and PFU microbial community in water samples had been investigated from seven sampling sites. The results indicated that the concentrations of Pb, Zn, Cd, Cu, and TSS in the mine drainage were gradually reduced from the inlet to the outlet of the wetland system and 99%, 98%, 75%, 83%, and 68% of these metals and TSS respectively, had been reduced in concentration after the drainage passed through the wetland system. A total of 105 protozoan species were identified, the number of protozoa species and the diversity index (DI) gradually increased, while the heterotrophic index (HI) gradually decreased from the inlet to the outlet of the wetland system. The results indicated that DI, HI, and total number species of protozoa could be used as biological indicators indicating the improvement of water quality.

Genetic heterogeneity and ploidy level analysis among different gynogenetic clones of the polyploid gibel carp

Background: Some triploid and tetraploid clones have been identified in the gynogenetic gibel carp, Carassius auratus gibelio Bloch, by karyotypic and cytologic analyses over many years. Further, 5-20% males and karyotypic diversity have been found among their natural and artificial populations. However, the DNA contents and the relation to their ploidy level and chromosome numbers have not been ascertained, and whether normal meiosis occurs in spermatogenesis needs to be determined in the different clones. Methods: The sampled blood cells or sperms were mixed with blood cells from chicken or individual gibel carp and fixed in 70% pre-cooled ethanol overnight at 4degreesC. The mixed cell pellets were washed 2-3 times in 1x phosphate buffered saline and then resuspended in the solution containing 0.5% pepsin and 0.1 M HCl. DNA was stained with propidium iodide solution (40 mug/mL) containing 4 kU/ml RNase. The measurements of DNA contents were performed with Phoenix Flow Systems. Results: Triploid clones A, E, F, and P had almost equal DNA content, but triploid clone D had greater DNA content than did the other four triploid clones. DNA content of clone M (7.01 +/- 0.15 pg/nucleus) was almost equal to the DNA content of clone D (5-38 +/- 0.06 pg/nucleus) plus the DNA content of common carp sperm (1.64 +/- 0.02 pg/nucleus). The DNA contents of sperms from clones A, P, and D were half of their blood cells, suggesting that normal meiosis occurs in spermatogenesis. Conclusions: Flow cytometry is a powerful method to analyze genetic heterogeneity and ploidy level among different gynogenetic clones of polyploid gibel carp. Through this study, four questions have been answered. (a) The DNA content correlation among the five triploid clones and one multiple tetraploid clone was revealed in the gibel carp, and the contents increased with not only the ploidy level but also the chromosome number. (b) Mean DNA content was 0.052 pg in six extra chromosomes of clone D, which was higher than that of each chromosome in clones A, E, F, and P (about 0.032 pg/ chromosome). This means that the six extra chromosomes are larger chromosomes. (c) Normal meiosis occurred during spermatogenesis of the gibel carp, because DNA contents of the sperms from clones A, P, and D were almost half of that in their blood cells. (d) Multiple tetraploid clone M (7.01 +/- 0.15 pg/nucleus) contained the complete genome of clone D (5.38 +/- 0.06 pg/nucleus) and the genome of common carp sperm (1.64 +/- 0.02 pg/nucleus). Cytometry Part A 56A:46-52, 2003. (C) 2003 Wiley-Liss, Inc.

Biological evaluation of running waters in Korea, with special reference to bioassessment using the PFU system

A total of six stations in the Han River system were selected for establishing polyurethane foam units (PFUs) to collect protozoans, including phytomastigophorans, zoomastigophorans, amoebas and ciliates, in July 1993. In the bioassessment of microbial communities using the PFUs, the number of species decreased as pollution intensity increased. The diversity index values calculated at the main stations generally agreed with the pollution status of the stations. Anyang-Chon (Chon means stream) showed the lowest diversity value (1.89), and all stations, except Masok and Anyang-Chon, showed diversity index values ranging from 3.15 to 3.93. The highest heterotrophic index (HI) value was detected in Anyang-Chon followed by Masok-Chon. The number of species at the main stations reached a maximum on day 11 of being exposed to PFUs. The results of S-eq, G and T-90% all suggest that bioassessments using the PFU system were well matched with pollution status of the water. All microbial variables were significantly correlated with comprehensive chemical pollution indices, P-a and P-b, with correlation coefficients ranging from r=0.87 to r=0.96.

Metamorphic InGaAs p-i-n Photodetectors with 1.75 mu m Cut-Off Wavelength Grown on GaAs

Top-illuminated metamorphic InGaAs p-i-n photodetectors (PDs) with 50% cut-off wavelength of 1.75 mu m at room temperature are fabricated on GaAs substrates. The PDs are grown by a solid-source molecular beam epitaxy system. The large lattice mismatch strain is accommodated by growth of a linearly graded buffer layer to create a high quality virtual InP substrate indium content in the metamorphic buffer layer linearly changes from 2% to 60%. The dark current densities are typically 5 x 10(-6) A/cm(2) at 0 V bias and 2.24 x 10(-4) A/cm(2) at a reverse bias of 5 V. At a wavelength of 1.55 mu m, the PDs have an optical responsivity of 0.48 A/W, a linear photoresponse up to 5 mW optical power at -4 V bias. The measured -3 dB bandwidth of a 32 mu m diameter device is 7 GHz. This work proves that InGaAs buffer layers grown by solid source MBE are promising candidates for GaAs-based long wavelength devices.

Impact of thickness of GaN buffer layer on properties of AlN/GaN distributed Bragg reflectors grown by metalorganic chemical vapor deposition

We studied the impact of the thickness of GaN buffer layer on the properties of distributed Bragg reflector (DBR) grown by metalorganic chemical vapor deposition (MOCVD). The samples were characterized by using metallographic microscope, transmission electron microscope (TEM), atomic force microscopy (AFM), X-ray diffractometer (XRD) and spectrophotometer. The results show that the thickness of the GaN buffer layer can significantly affect the properties of the DBR structure and there is an optimal thickness of the GaN buffer layer. This work would be helpful for the growth of high quality DBR structures.

Observation of the surface circular photogalvanic effect in InN films

A sizable spin-dependent photocurrent related to the interband transition in InN films is observed. The surface charge accumulation layer is suggested to be the origin of the circular photogalvanic current, which is consistent with the result of uniaxial strain experiments and the comparison of front and back incidence. The homogeneous photocurrent demonstrates the existence of spin splitting in the InN surface layer, and the structure inversion asymmetry (SIA)-dominant mechanism indicates a great possibility for the manipulation of spin splitting, which would undoubtedly benefit further research and applications of spintronics. Crown Copyright (C) 2009 Published by Elsevier Ltd. All rights reserved.

Surface characteristics of SiO2-TiO2 strip fabricated by laser direct writing

SiO2-TiO2 sol-gel films are deposited on SiO2/Si by dip-coating technique. The SiO2-TiO2 strips are fabricated by laser direct writing using all ytterbium fiber laser and followed by chemical etching. Surface structures, morphologies and roughness of the films and strips are characterized. The experimental results demonstrate that the SiO2-TiO2 sol-gel film is loose in Structure and a shrinkage concave groove forms if the film is irradiated by laser beam. The surface roughness of both non-irradiated and laser irradiated areas increases with the chemical etching time. But the roughness of laser irradiated area increases more than that of non-irradiated area under the same etching time. After being etched for 28 s, the surface roughness value of the laser irradiated area increases from 0.3 nm to 3.1 nm.