Short and long peptidoglycan recognition proteins (PGRPs) in zebrafish, with findings of multiple PGRP homologs in teleost fish

Peptidoglycan recognition protein (PGRP) specifically binds to peptidoglycan and is considered to be one of the pattern recognition proteins in the innate immunity of insect and mammals. Using a database mining approach and RT-PCR, multiple peptidoglycan recognition protein (PGRP) like genes have been discovered in fish including zebrafish Danio rerio, Japanese pufferfish TakiFugu rubripes and spotted green pufferfish Tetraodon nigroviridis. They share the common features of those PGRPs in arthropod and mammals, by containing a conserved PGRP domain. Based on the predicted structures, the identified zebrafish PGRP homologs resemble short and long PGRP members in arthropod and mammals. The identified PGRP genes in T. nigroviridis and TakiFugu rubripes resemble the long PGRPs, and the short PGRP genes have not been found in T. nigroviridis and TakiFugu rubripes databases. Computer modelling of these molecules revealed the presence of three alpha-helices and five or six beta-strands in all fish PGRPs reported in the present study. The long PGRP in teleost fish have multiple alternatively spliced forms, and some of the identified spliced variants, e.g., tnPGRP-L3 and tnPGRP-L4 (in: Tetraodon nigroviridis), exhibited no characters present in the PGRP homologs domain. The coding regions of zfPGRP6 (zf: zebrafish), zfPGRP2-A, zfPGRP2-B and zfPGRP-L contain five exons and four introns; however, the other PGRP-like genes including zfPGRPSC1a, zfPGRPSC2, tnPGRP-L1-, tnPGRP-L2 and frPGRP-L (fr: Takifugu rubripes) contain four exons and three introns. In zebrafish, long and short PGRP genes identified are located in different chromosomes, and an unknown locus containing another long PGRP-like gene has also been found in zebrafish, demonstrating that multiple PGRP loci may be present in fish. In zebrafish, the constitutive expressions of zfPGRP-L, zfPGRP-6 and zfPGRP-SC during ontogeny from unfertilized eggs to larvae, in different organs of adult, and the inductive expression following stimulation by Flavobacterium columnare, were detected by real-time PCR, but the levels and patterns varied for different PGRP genes, implying that different short and long PGRPs may play different roles in innate immune response. (c) 2007 Elsevier Ltd. All rights reserved.

Retention of the developmental pluripotency in medaka embryonic stem cells after gene transfer and long-term drug selection for gene targeting in fish

Embryonic stem (ES) cells provide a unique tool for introducing random or targeted genetic alterations, because it is possible that the desired, but extremely rare recombinant genotypes can be screened by drug selection. ES cell-mediated transgenesis has so far been limited to the mouse. In the fish medaka (Oryzias latipes) several ES cell lines have been made available. Here we report the optimized conditions for gene transfer and drug selection in the medaka ES cell line MES1 as a prelude for gene targeting in fish. MES1 cells gave rise to a moderate to high transfection efficiency by the calcium phosphate co-precipitation (5%), commercial reagents Fugene (11%), GeneJuice (21%) and electroporation (>30%). Transient gene transfer and CAT reporter assay revealed that several enhancers/promoters and their combinations including CMV, RSV and ST (the SV40 virus early gene enhancer linked to the thymidine kinase promoter) were suitable regulatory sequences to drive transgene expression in the MES1 cells. We show that neo, hyg or pac conferred resistance to G418, hygromycin or puromycin for positive selection, while the HSV-tk generated sensitivity to ganciclovir for negative selection. The positive-negative selection procedure that is widely used for gene targeting in mouse ES cells was found to be effective also in MES1 cells. Importantly, we demonstrate that MES1 cells after gene transfer and long-term drug selection retained the developmental pluripotency, as they were able to undergo induced differentiation in vitro and to contribute to various tissues and organs during chimeric embryogenesis.

Spawning areas and early development of long spiky-head carp (Luciobrama macrocephalus) in the Yangtze River and Pearl River, China

The spawning areas and early development of long spiky-head carp, Luciobrama macrocephalus (Lacepede), an endemic fish species in China, were investigated in the Yangtze River and Pearl River of central and southeastern China between 1961 and 1993. The potamodromous fish migrated upstream to spawn between May and July as the floodwater began to rise. The water-hardened eggs drifted down the river, and the embryos and larvae developed in the course of drifting. The spawning areas of the fish were widely found in the upper and middle main channels and large tributaries. Two large dams (Gezhouba dam and Danjiangkou dam) did not significantly impact on the reproduction of the fish. Fifty stages of the early development from one cell to the juvenile with fully formed fins were observed and characterized pictorially. The larvae of long spiky-head carp could be distinguished from the larvae of other co-occurring species by counting the number of somites and comparing the proportion of sizes of eye to otic capsule.

Long-term (1956-1999) dynamics of phosphorus in a shallow, subtropical Chinese lake with the possible effects of cyanobacterial blooms

This paper describes the long-term dynamics of phosphorus concentrations in both the lake water and the sediment in a subtropical Chinese lake, Lake Donghu. The total phosphorus (TP) concentration in the lake water experienced an upward trend from the 1950s, and peaked in 1983/1984, but declined obviously afterwards. From the 1950s to the 1990s, TP content in the upper 10 cm sediment of the lake increased steadily from 0.307 to 1.68 mg Pg DW-1 at Station I and from 0.151 to 0.89 mg Pg DW-1 at Station II, respectively. The TP increase in the lake water before mid-1980s was mainly attributed to the massive input of sewage P. The outbreak of cyanobacterial blooms coincided with the peaks of TP and Orthophosphate (PO4-P) in the water in mid-1980s, and the maximum TP of the water reached as high as 1.349 mg/1 at Station I and 0.757 mg/l at Station II (in 1984), respectively. The declines of TP and PO4-P in the water after mid-1980s was coincident with the disappearance of cyanobacterial bloom. (C) 2001 Elsevier Science Ltd. All rights reserved.

Long-term changes of planktonic rotifers in a subtropical Chinese lake dominated by filter-feeding fishes

1. The long-term changes (1956-1998) in density and species composition of planktonic rotifers were studied at two sampling stations (I, II) of Lake Donghu, a shallow eutrophic Chinese Lake densely stocked with filter-feeding fishes. Annual average densities of rotifers increased with an increase in fish yield and eutrophication, whilst species number decreased from 82 in 1962-1963 to 62 in 1994-1998. 2. During 1962-98, some species such as Anuraeopsis fissa, Polyarthra spp. (including P. dolichoptera & P. vulgaris), Trichocerca pusilla and Synchaeta oblonga increased their percentage in abundance remarkably, whilst the proportion of Keratella cochlearis decreased at two relatively eutrophic stations from 19 to 4.2% at Station I and from 30 to 3.2% at Station IL 3. The high r(max) of A. fissa probably made it more successful than other rotifers under high predation pressure by planktivorous fish. The decrease in the K. cochlearis population might be attributed partly to predation by Cyclops vicinus. 4. Small rotifers were less vulnerable to fish predation than large-sized cladocerans. Decreases in cladocerans coincided with increases in rotifers, suggesting that the indirect effect of fish predation on cladocerans might have partly contributed to the population development of rotifers in Lake Donghu during recent decades. 5. We also conducted surveys (1994-1998) of seasonal dynamics of rotifers at four sampling stations (I-IV) which have varied in trophic status after fragmentation of the lake in the 1960s. A total of 75 species were identified at the four stations. Both densities and biomass of rotifers were considerably higher in the two more eutrophic stations than in the two less eutrophic stations. This indicates that the population increase of rotifers at Stations I and II during recent decades might be partly attributed to eutrophication of the lake water.

Impacts of filter-feeding fishes on the long-term changes of crustacean zooplankton in a eutrophic subtropical Chinese lake

Long-term changes In the crustacean zooplankton community (calanoid and cyclopoid copepods and cladocerans) were studied in Lake Donghu, a shallow and eutrophic Chinese lake. This lake had been earlier stocked with two pump Alter-feeding Ashes, silver carp (Hypopthalmichthys molitrix) and bighead carp (Aristichthys nobilis). During the 1950s and the mid-1980s, the ratio of copepods to cladocerans was relatively stable but showed a general increase thereafter. From the early-1980s to the 1990s, calanoid/cyclopoid ratios decreased obviously. In the 1990s, Cyclops vicinus, Diaphanosoma brachyurum, and Moina micrura were dominant the abundance of C. vicinus and M. micrura increased significantly; and D, brachyurum showed a substantial decrease. The study shows that under extremely high pressure of Ash predation, the species which could recover rapidly from fish predation would be the most likely to survive and increase their numbers.

Long-term toxic impact of 2,3,7,8-tetrachlorodibenzo-p-dioxin on the reproduction, sexual differentiation, and development of different life stages of Gobiocypris rarus and Daphnia magna

The sexual ratio of Gobiocypris rarus exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and 17 beta -estradiol from embryo to sexually mature revealed feminization and overdevelopment of connective tissue in male fish gonad in 2-30 pg/L TCDD concentration range. Daphnia magna was not sensitive to the high dose of TCDD (0.1-1000 ng/ml), but the reproduction of D. magna treated with TCDD decreased after the 8th day. 7-Ethoxyresorufin-O-deethylase (EROD) activities in newly fertilized eggs of G. rarus exposed to TCDD dosage groups (1000-100,000 pg/L) were significantly induced and increased with TCDD concentrations at the early life stage, while no difference was found between low TCDD dosage groups (<100 pg/L), but a good relationship between the EROD activity and the TCDD concentration was observed during a long-term developmental stage. There was a pericardial edema formed in a 2-week yolk-sac at the concentration of 1000 pg/L TCDD. But in the exposure group (2 pg/L TCDD for 120 days), the cell nuclei of hepatocytes was far from the center and packed toward the cell membrane; the cristae of most mitochondria in the cell dropped and collapsed; the rough endoplasmic reticulum broke into fragments; and numerous lipid droplets formed in the cell. (C) 2001 Academic Press.

Long-term changes of Copepoda community (1957-1996) in a subtropical Chinese lake stocked densely with planktivorous filter-feeding silver and bighead carp

In contrast to the relatively well documented impact of particulate-feeding fish on zooplankton communities, little attention has been devoted to the impact of filter-feeding fish. Filter-feeding silver and bighead carp are the most intensively cultured fish species in Asia and comprise much of the production of Chinese aquaculture. However, little information is known about the impact of either fish on the zooplankton community. Long-term changes in the Copepoda community (1957-1996) were studied at two sampling stations of a subtropical Chinese lake (Lake Donghu) dominated by silver and bighead carp. For both calanoids and cyclopoids, the littoral station (I) was much more resource profitable than the pelagic station (II). There has been a tremendous increase in the annual fish catch over the past 30 years due to the increased stocking with fingerlings of the two carp species. There was a notably higher fish density at Station I than at Station II. Cyclopoid abundance was notably higher at Station I than at Station II during the 1950s to the 1980s, while the reverse became true in the 1990s. This is probably because when fish abundance increased to an extremely high level, the impact of fish predation on the cyclopoids became more important than that of food resources at the littoral station. At both stations, cyclopoid abundance was relatively low in spite of the presence of abundant prey. Similarly, calanoid density did not differ significantly between the two stations in the 1950s and 1960s, but was significantly lower at Station I than at Station II during the 1980s and 1990s. Such changes are attributed to the gradient of fish predation between the stations and an increasing predation pressure by the fish. The increased fish predation also correlated with a shift in summer-dominant calanoids from larger species to smaller ones. In conclusion, the predaceous cyclopoids are affected by fish predation to a much lesser extent than the herbivorous calanoids, and therefore increased predation by filter-feeding fish results in a definite increase in the cyclopoid/calanoid ratio. Predation by filter-feeding fish has been a driving force in shaping the copepod community structure of Lake Donghu during the past decades.

Distribution, transformation, and long-term accumulation of polychlorinated dibenzo-p-dioxins and dibenzofurans in different tissues of fish and piscivorous birds

The present study monitored 10-year-old fish and piscivorous birds from sites contaminated for many Stars. The data reflected the results of actual, long-term environmental exposures, The results demonstrate that different tissues of fish have quite different concentrations of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/F), The concentration order of PCDD/F within fish is liver congruent to egg congruent to intestines kidney congruent to hearts gill congruent to bladders > muscle > brain. The concentration order of PCDD/F within piscivorous birds was livers egg congruent to hearts muscle congruent to stomachs brain, The results obtained also demonstrate that the accumulation patterns of piscivorous birds and fish are quite different. The tissues of fish and piscivorous birds have different capacities for bioaccumulation and biotransformation of PCDD/F; variable proportions of TEQs were also found throughout their bodies. In fish, toxic equivalency quotient (TEQ): PCDD/F ratios in various tissues ranged from 0.01 to 0.07, whereas in birds the ratios ranged from 0.07 to 0.43. If the concentrations are normalized with lipid content, the results vary less. The effect of different lipid properties is obvious in the case of brain tissue, which is richer in phospholipids. (C) 2000 Academic Press.

Long-term changes in crustacean zooplankton and water quality in a shallow, eutrophic Chinese lake densely stocked with fish

From surveys made in 1962-1963, 1973-1974, 1979-1996 at two Stations in Lake Donghu, a shallow eutrophic water body near Wuhan, P. R. China, the authors, derive long-term changes in species composition, standing crop and body-size of planktonic crustaceans. The species number decreased from the 1960s to the 1990s. The cladocerans dropped from 46 (1960s) to 26 (1980s) to 13 (1990s); the copepods decreased from 14 (1960s) to 10 (1980s) to 7 (1990s). From the mid-1980s on, the dominant crustaceans also changed: Daphnia hyalina and D. carinata ssp. were replaced by Moina micrura and Diaphanosoma brachyurum at Stations 1 and 2, respectively; Cyclops vicinus replaced Mesocyclops leuckarti. Densities and biomass of Cladocera decreased markedly after 1987. Annual average densities and biomass of cladocerans were statistically differences between 1962-1986 and 1987-1996 (P < 0.01). Annual average densities of Daphnia (Station 1 + Station 2) were negatively correlated with fish yield. Since the 1980s, annual average body length of Cladocera and Calanoida decreased, while annual average body length of Cyclopoida increased. In the same years, average body length of copepods was lower during May-October than during January-April and November-December. A 12-yr data analysis showed annual average concentration of chlorophyll-a (Chl-a) to be negatively correlated with annual average density of Daphnia, whilst lake transparency was positively correlated with annual average densities of Daphnia. The results imply that, since Daphnia feeds efficiently on phytoplankton, it could decrease concentration of Chl-a, and enhance water transparency.

Long rectangle resonator 1550 nm AlGaInAs/InP lasers

1550 nm AlGaInAs/InP long rectangle resonator lasers with three sides surrounded by SiO2 and p electrode layers are fabricated by planar technology, and room-temperature continuous-wave lasing is realized for a laser with a length of 53 mu m and a width of 2 mu m. Multiple peaks with wavelength intervals of Fabry-Perot mode intervals and mode Q factors of about 400 and a lasing mode with a Q factor over 8000 are observed from the lasing spectrum at threshold current. The numerical results of the FDTD simulation indicate that the lasing mode may be a whispering-gallery mode, which is a coupled mode of two high-order transverse modes of the waveguide.

Fracture Properties of LPCVD Silicon Nitride and Thermally Grown Silicon Oxide Thin Films From the Load-Deflection of Long Si3N4 and SiO2/Si3N4 Diaphragms

The bulge test is successfully extended to the determination of the fracture properties of silicon nitride and oxide thin films. This is achieved by using long diaphragms made of silicon nitride single layers and oxide/nitride bilayers, and applying comprehensive mechanical model that describes the mechanical response of the diaphragms under uniform differential pressure. The model is valid for thin films with arbitrary z-dependent plane-strain modulus and prestress, where z denotes the coordinate perpendicular to the diaphragm. It takes into account the bending rigidity and stretching stiffness of the layered materials and the compliance of the supporting edges. This enables the accurate computation of the load-deflection response and stress distribution throughout the composite diaphragm as a function of the load, in particular at the critical pressure leading to the fracture of the diaphragms. The method is applied to diaphragms made of single layers of 300-nm-thick silicon nitride deposited by low-pressure chemical vapor deposition and composite diaphragms of silicon nitride grown on top of thermal silicon oxide films produced by wet thermal oxidation at 950 degrees C and 1050 degrees C with target thicknesses of 500, 750, and 1000 mn. All films characterized have an amorphous structure. Plane-strain moduli E-ps and prestress levels sigma(0) of 304.8 +/- 12.2 GPa and 1132.3 +/- 34.4 MPa, respectively, are extracted for Si3N4, whereas E-ps = 49.1 +/- 7.4 GPa and sigma(0) = -258.6 +/- 23.1 MPa are obtained for SiO2 films. The fracture data are analyzed using the standardized form of the Weibull distribution. The Si3N4 films present relatively high values of maximum stress at fracture and Weibull moduli, i.e., sigma(max) = 7.89 +/- 0.23 GPa and m = 50.0 +/- 3.6, respectively, when compared to the thermal oxides (sigma(max) = 0.89 +/- 0.07 GPa and m = 12.1 +/- 0.5 for 507-nm-thick 950 degrees C layers). A marginal decrease of sigma(max) with thickness is observed for SiO2, with no significant differences between the films grown at 950 degrees C and 1050 degrees C. Weibull moduli of oxide thin films are found to lie between 4.5 +/- 1.2 and 19.8 +/- 4.2, depending on the oxidation temperature and film thickness.

Fracture properties of silicon carbide thin films by bulge test of long rectangular membrane

This paper reports the mechanical properties and fracture behavior of silicon carbide (3C-SiC) thin films grown on silicon substrates. Using bulge testing combined with a refined load-deflection model of long rectangular membranes, which takes into account the bending stiffness and prestress of the membrane material, the Young's modulus, prestress, and fracture strength for the 3C-SiC thin films with thicknesses of 0.40 and 1.42 mu m were extracted. The stress distribution in the membranes under a load was calculated analytically. The prestresses for the two films were 322 +/- 47 and 201 +/- 34 MPa, respectively. The thinner 3C-SiC film with a strong (111) orientation has a plane-gstrain moduli of 415 +/- 61 GPa, whereas the thicker film with a mixture of both (111) and (110) orientations exhibited a plane-strain moduli of 329 +/- 49 GPa. The corresponding fracture strengths for the two kinds of SiC films were 6.49 +/- 0.88 and 3.16 +/- 0.38 GPa, respectively. The reference stresses were computed by integrating the local stress of the membrane at the fracture over edge, surface, and volume of the specimens and were fitted with Weibull distribution function. For the 0.40-mu m-thick membranes, the surface integration has a better agreement between the data and the model, implying that the surface flaws are the dominant fracture origin. For the 1.42-mu m-thick membranes, the surface integration presented only a slightly better fitting quality than the other two, and therefore, it is difficult to rule out unambiguously the effects of the volume and edge flaws.

Spectroscopy of long wavelength coupled quantum dots

We obtained a low density of coupled InAs/GaAs quantum dots (QDs) with an emission wavelength of around 1.3 mu m at room temperature. Atomic force microscopy and transmission electronic microscopy reveal that the dot size difference and the lateral displacement between the two dots are related to the spacer thickness. Spectroscopy of the coupled QD ensembles is considerably influenced by the spacer thickness.

Long-wavelength light emission from self-assembled heterojunction quantum dots

The authors report the optical characteristics of GaSb/InAs/GaAs self-assembled heterojunction quantum dots (QDs). With increasing GaSb deposition, the room temperature emission wavelength can be extended to 1.56 mu m. The photoluminescence mechanism is considered to be a type-II transition with electrons confined in InAs and holes in GaSb.(C) 2008 American Institute of Physics.