A Dicer-1 gene from white shrimp Litopenaeus vannamei: Expression pattern in the processes of immune response and larval development

Dicer is a member of the RNAase III family which catalyzes the cleavage of double-stranded RNA to small interfering RNAs and micro RNAs, and then directs sequence-specific gene silencing. In this paper, the full-length cDNA of Dicer-1 was cloned from white shrimp Litopenaeus vannamei (designated as LvDcr1). It was of 7636 bp, including a poly A tail, a 5' UTR of 136 bp, a 3' UTR of 78 bp, and an open reading frame (ORF) of 7422 bp encoding a putative protein of 2473 amino acids. The predicted amino acid sequence comprised all recognized functional domains found in other Dicer-1 homologues and showed the highest (97.7%) similarity to the Dicer-1 from tiger shrimp Penaeus mondon. Quantitative real-time PCR was employed to investigate the tissue distribution of LvDcr1 mRNA, and its expression in shrimps under virus challenge and larvae at different developmental stages. The LvDcr1 mRNA could be detected in all examined tissues with the highest expression level in hemocyte, and was up-regulated in hemocytes and gills after virus injection. These results indicated that LvDcr1 was involved in antiviral defense in adult shrimp. During the developmental stages from fertilized egg to postlarva VII, LvDcr1 was constitutively expressed at all examined development stages, but the expression level varied significantly. The highest expression level was observed in fertilized eggs and followed a decrease from fertilized egg to nauplius I stage. Then, the higher levels of expression were detected at nauplius V and postlarva stages. LvDcr1 expression regularly increased at the upper phase of nauplius, zoea and mysis stages than their prophase. The different expression of LvDcr1 in the larval stages could provide clues for understanding the early innate immunity in the process of shrimp larval development. (C) 2010 Elsevier Ltd. All rights reserved.

Broader pattern of tandem repeats in the mitochondrial control region of Perciformes

Perciformes, the largest order of vertebrates with 20 suborders, is the most diverse fish order that dominates vertebrate ocean life. The complete mitochondrial control region (CR) of Trichiurus japonicus (Trichiuridae, Scombroidei) and Pampus sp. (Stromateidae, Stromateoidei) were amplified and sequenced. Together with data from GenBank, the tandem repeats in the mitochondrial CR from 48 species, which covered nine suborders of Perciformes, are reported in this study. The tandem repeats tend to be long in the suborder Percoidei and Stromateoidei. The identical repeats in 21 species of Cichlidae suggest a common origin and have existed before species divergence. Larimichthys crocea shows tandem repeats instead of the typical structure of the central conserved sequence blocks, which was first reported in Perciformes and vertebrates. This might have resulted from interruption of the polymerase activity during the H-strand synthesis. The four broader patterns presented here for the tandem repeats, including those in both the 5' and 3' ends, only in the either 5' or 3' end, and in the central conserved domain of the control region, will be useful for understanding the evolution of species.

CfLGBP, a pattern recognition receptor in Chlamys farreri involved in the immune response against various bacteria

Lipopolysaccharide and beta-1, 3-glucan binding protein (LGBP) is a kind of pattern recognition receptor, which can recognize and bind LPS and beta-1, 3-glucan, and plays curial roles in the innate immune defense against Gram-negative bacteria and fungi. In this study, the functions of LGBP from Zhikong scallop Chlamys farreri performed in innate immunity were analyzed. Firstly, the mRNA expression of CfLGBP in hemocytes toward three typical PAMPS stimulation was examined by realtime PCR. It was up-regulated extremely (P < 0.01) post stimulation of LPS and beta-glucan, and also exhibited a moderate up-regulation (P < 0.01) after PGN injection. Further PAMPs binding assay with the polyclonal antibody specific for CfLGBP proved that the recombinant CfLGBP (designated as rCfLGBP) could bind not only LPS and beta-glucan, but also PGN in vitro. More importantly, rCfLGBP exhibited obvious agglutination activity towards Gram-negative bacteria Escherichia coil, Gram-positive bacteria Bacillus subtilis and fungi Pichia pastoris. Taking the results of immunofluorescence assay into account, which displayed CfLGBP was expressed specifically in the immune cells (hemocytes) and vulnerable organ (gill and mantle), we believed that LGBP in C farreri, serving as a multi-functional PRR, not only involved in the immune response against Gram-negative and fungi as LGBP in other invertebrates, but also played significant role in the event of anti-Gram-positive bacteria infection. As the first functional research of LGBP in mollusks, our study provided new implication into the innate immune defense mechanisms of C. farreri and mollusks. (C) 2010 Elsevier Ltd. All rights reserved.

Cflec-5, a pattern recognition receptor in scallop Chlamys farreri agglutinating yeast Pichia pastoris

C-type lectins are a superfamily of carbohydrate-recognition proteins which play crucial roles as pattern recognition receptors (PRRs) in the innate immunity. In this study, the full-length cDNA of a C-type lectin was cloned from scallop Chlamys farreri (designated as Cflec-5) by expression sequence tag (EST) analysis and rapid amplification of cDNA ends (RACE) approach The full-length cDNA of Cflec-5 was of 1412 bp. The open reading frame encoded a polypeptide of 153 amino acids, including a signal sequence and a conserved carbohydrate-recognition domain with the EPN motif determining the mannose-binding specificity The deduced amino acid sequence of Cflec-5 showed high similarity to members of C-type lectin superfamily. The quantitative real-time PCR was performed to investigate the tissue distribution of Cflec-5 mRNA and its temporal expression profiles in hemocytes post pathogen-associated molecular patterns (PAMPs) stimulation. In healthy scallops, the Cflec-5 mRNA was mainly detected in gill and mantle, and marginally in other tissues The mRNA expression of Cflec-5 could be significantly induced by lipopolysaccharide (LPS) and glucan stimulation and reached the maximum level at 6 h and 12 h, respectively But its expression level did not change significantly during peptidoglycan (PGN) stimulation The function of Cflec-5 was investigated by recombination and expression of the cDNA fragment encoding its mature peptide in Escherichia coli Rosetta Gami (DE3) The recombinant Cflec-5 agglutinated Pichia pastoris in a calcium-independent way The agglutinating activity could be inhibited by D-mannose. LPS and glucan, but not by D-galactose or PGN. These results collectively suggested that Cflec-5 was involved in the innate Immune response of scallops and might contribute to nonself-recognition through its interaction with various PAMPs (C) 2010 Elsevier Ltd All rights reserved

Tectonic and geodynamic pattern of marginal seas on the West Pacific inferred from multi-satellite altewetry geoid

With the rapid development of satellite observations, we can use the altimetry geoid to study submarine tectonics and geodynamics. On the basis of the 4' x 4' geoid undulation calculated from altimeter data of Geosat, ERS-1/2 and Topex/Poseidon on the West Pacific, located between 0degreesN similar to 45degreesN, 100degreesC similar to 150degreesE, Bouguer, Glenni and isostatic geoid undulation are obtained from correction of gravitational potential of the global topography and isostacy. Moho discontinuity depth is inversed by the Glenni geoid undulation, and the stress field from small-scale mantle convection is reasonably calculated from the isostatic geoid undulation. The results show that within the Philippine Sea and the South China Sea, short-wavelength lineations of the geoid undulation are parallel or cross to magnetic lineations and rifting ridges. The Moho depth of marginal sea basins becomes shallow southward, and its values are similar to that of the Philippine Sea. These facts show that strength of tectonic activities are almost the same on the both sides of the Ryukyu-Taiwan-Philippine are. Various kinds of tectonic features with different driving mechanisms of small-middle and large-scale of mantle convection, however, display a special pattern of tectonics and geodynamics of the continental marginal seas distinguished from oceans and continents.

Distribution pattern of marine flagellate and its controlling factors in the central and north part of the Huanghai Sea in early summer

A survey was carried out in the central and north part of the Huanghai Sea (34.5degrees similar to 37.0degreesN, 120.5degrees similar to124.0degreesE) during June 12 similar to 27, 2000. It was found that the abundance of marine flagellate ranged from 45 to 1278 cell/ml, 479 cell/ml in average. Flagellate was more abundant in the central part than in the north part of Huanghai Sea, and the abundance decreased with the increasing distance from the coast, showing a similar distribution pattern with isotherm. Vertically, high density of flagellate was always presented in the bottom of thermocline, and formed a dense accumulation in the central area of the Huanghai Sea Cold Water Mass. The effects of physical and biological factors on the distribution of marine flagellate in early summer were discussed. Water temperature (especially the existence of thermocline) rather than salinity showed significant effect on the distribution pattern of marine flagellate in the Huanghai Sea in early summer. When comparing the abundance of marine flagellate with that of other microorganisms, it revealed a comparatively stable relationship among these organhisms, with a ratio of heterotrophic bacteria: cyanobacteria: flagellate: dinoflagellate: ciliate being 10(5) 10(3):10(2):10(1):10(0).

Preliminary study of the dynamic origin of the distribution pattern of bottom sediments on the continental shelves of the Bohai Sea, Yellow Sea and East China Sea

The bottom sediment types in the Bohai Sea, Yellow Sea and East China Sea (BYECS) are diversified, and their distribution pattern is very complicated. However, the bottom sediment types can be simplified to be sandy sediment, clayey sediment and mixed sediment, which comprise the complicated distribution pattern of bottom sediment in the BYECS. The continental shelves of the BYECS are broad, with shallow water depths and tidal currents which are permanent and dominate the marine dynamics in the BYECS. Based on numerical simulation of tidal elevations and currents in the BYECS, the rates of suspended load transport and bed load transport during a single tidal cycle for sediments of eight different grain size ranges are calculated. The results show that any sediment, whose threshold velocity is less than that of tidal current, has the same transport trend. Suspended load transport rare, bed load transport rate, and the ratio of the former to the latter decrease with grain size becoming coarser and coarser. The erosion/accretion patterns of sediments with different grain sizes are determined by the sediment transport rate divergences, and the results show that the patterns are the same for sediments with different grain sizes. Three main bottom sediment types, i.e. sandy sediment mainly composed of fine sand, clayey sediment mainly composed of silty clay, and mixed sediment mainly composed of fine sand, silt, and clay, are obtained by computation. The three bottom sediment types and their distribution pattern are consistent not only with sediment transport field and the sea bed erosion/accretion pattern obtained by simulation, but also with field data of bottom sediment types and divisions. In the BYECS, sand ridges form mainly in the areas with strong rectilinear tidal currents, sand sheets form mainly in the areas dominated by strong rotatory tidal currents, and clayey sediments, i.e. mud patches, form mainly in the areas with weak tidal currents. Hence, not only the sandy sediments but also the clayey sediments in the BYECS are formed under the control of the whole tidal current field of the BYECS. The three main bottom sediment types are not isolated respectively-in fact, they constitute a whole tidal depositional system. Under the condition with no cyclonic cold eddy, the clayey sediments in the BYECS can form in weak tidal current environments. Therefore, a cold eddy is not necessary for the deposition of clayey sediments in the BYECS. (C) 2000 Academic Press.

Studies on the transmission of WSSV (white spot syndrome virus) in juvenile Marsupenaeus japonicus via marine microalgae

We studied the possible role that marine microalgae may play during the outbreaks of WSS (white spot syndrome). In order to elucidate the possibility of marine microalgae carrying WSSV (white spot syndrome virus), six marine microallgae (Isochr.vsis galbana, Skeletonema costatum, Chlorella sp., Heterosigma akashiwo, Scrippsiella trochoidea, Dunaliella salina) were co-cultured with adult Marsupenaeus japollicus infected with WSSV and were assayed daily by nested-PCR to study whether they could carry WSSV. Further experiments were conducted to investigate whether the virus carried by microalgae could re-infect juvenile M. japonicus. Results showed that all of the experimental microalgae, except H. akashiwo could carry WSSV, and among them, Chlorella sp. and S. trochoidea had the strongest WSSV-carrying ability. Unlike other invertebrate carriers of WSSV, the WSSV detections in microalgae, which were positive after I and 3 days, were negative after 10 days of incubation. WSSV detection results in juvenile M. japonicus showed that the juvenile shrimp were re-infected by co-cultured Chlorella sp., although the juvenile M. japonicus carried so small an amount of WSSV that it could only be detected by nested-PCR. The results of this experiment suggest that microalgae might be one possible horizontal transmission pathway for WSSV. Further research, however, is required to better understand the factors behind the different carrying abilities and virus-carrying mechanisms of different microalgae. (c) 2007 Elsevier Inc. All rights reserved.

大型反倾岩体变形破坏机制及稳定性分析———以金沙江宽谷坝址龙蟠边坡为例

A large number of catastrophic accidents were aroused by the instability and destruction of anti-dip rock masses in the worldwide engineering projects, such as hydropower station, mine, railways and so on. Problems in relation to deformation and failure about anti-dip rock slopes are significant for engineering geology research. This dissertation takes the Longpan slope in the Jinsha River as a case to study the deformation mechanism of large-scale anti-dip rock masses and the slope stability analysis method. The primary conclusions are as follows. The Dale Reach of Jinsha River, from Longpan to the debouchment of Chongjiang tributary, is located in the southeastern margin of the Qinghai-Tibet Plateau. Longpan slope is the right embankment of Dale dam, it is only 26 km to the Shigu and 18 km to Tiger Leaping Gorge. The areal geology tectonic structures here area are complicated and blurry. Base on the information of geophysical exploration (CSAMT and seismology) and engineering geological investigation, the perdue tectonic pattern of Dale Reach is put forward for the first time in this paper. Due to the reverse slip of Longpan fault and normal left-rotation of Baihanchang fault, the old faulted valley came into being. The thick riverbed sediments have layered characters of different components and corresponding causes, which attribute to the sedimentary environments according with the new tectonic movements such as periodic mountain uplifting in middle Pleistocene. Longpan slope consists of anti-dip alternate sandstone and slate stratums, and the deformable volume is 6.5×107m3 approximately. It was taken for an ancient landslide or toppling failure in the past so that Dale dam became a vexed question. Through the latest field surveying, displacement monitoring and rock masses deforming characters analyses, the geological mechanism is actually a deep-seated gravitational bending deformation. And then the discrete element method is used to simulate the deforming evolution process, the conclusion accords very well with the geo-mechanical patterns analyses. In addition strength reduction method based on DEM is introduced to evaluate the factor of safety of anti-dip rock slope, and in accordance with the expansion way of the shear yielding zones, the progressive shear failure mechanism of large-scale anti-dip rock masses is proposed for the first time. As an embankment or a close reservoir bank to the lower dam, the stability of Longpan slope especially whether or not resulting in sliding with high velocity and activating water waves is a key question for engineering design. In fact it is difficult to decide the unified slip surface of anti-dip rock slope for traditional methods. The author takes the shear yielding zones acquired form the discrete element strength reduction calculation as the potential sliding surface and then evaluates the change of excess pore pressure and factor of stability of the slope generated by rapid drawdown of ponded water. At the same time the dynamic response of the slope under seismic loading is simulated through DEM numerical modeling, the following results are obtained. Firstly the effective effect of seismic inertia force is resulting in accumulation of shear stresses. Secondly the discontinuous structures are crucial to wave transmission. Thirdly the ultimate dynamic response of slope system takes place at the initial period of seismic loading. Lastly but essentially the effect of earthquake load to bringing on deformation and failure of rock slope is the coupling effect of shear stresses and excess pore water pressure accumulation. In view of limitations in searching the critical slip surface of rock slope of the existing domestic and international software for limit equilibrium slope stability analyses, this article proposes a new method named GA-Sarma Algorithm for rock slope stability analyses. Just as its name implies, GA-Sarma Algorithm bases on Genetic Algorithm and Sarma method. GA-Sarma Algorithm assumes the morphology of slip surface to be a broken line with traceability to extend along the discontinuous surface structures, and the slice boundaries is consistent with rock mass discontinuities such as rock layers, faults, cracks, and so on. GA-Sarma Algorithm is revolutionary method that is suitable for global optimization of the critical slip surface for rock slopes. The topics and contents including in this dissertation are closely related to the difficulties in practice, the main conclusions have been authorized by the engineering design institute. The research work is very meaningful and useful for the engineering construction of Longpan hydropower station.

条件反射性免疫抑制及其脑机制的c-fos研究

To explore the neural mechanisms underlying conditioned immunomodulation, this study employed the classical taste aversion (CTA) behavioral paradigm to establish the conditioned humoral and cellular immunosuppression (CIS) in Wistar rats, by paring saccharin (CS) with intraperitoneal (i.p.) injection of an immunosuppressive drug cyclophophamide (UCS). C-fos immunohistochemistry method was used to observe the changes of the neuronal activities in the rat brain during the acquisition, expression and extinction of the conditioned immunosuppression (CIS). The followings are the main results: 1. Five days after one trial of CS-UCS paring, reexposure to CS alone significantly decreased the level of the anti-ovalbumin (OVA) IgG in the peripheral serum. Two trials of CS-UCS paring and three reexposures to CS not only resulted in further suppression of the primary immune response, but also reduced the numbers of peripheral lymphocytes and white blood cells. This finding indicates that CS can induce suppression of the immune function, and the magnitude of the effects is dependent on the intensity of training. 2. On day 5 following two trials of CS-UCS pairing, CS suppressed the spleen lymphocytes responsiveness to mitogens ConA, PHA and PWM, and decreased the numbers of peripheral lymphocytes and white blood cells. On day 15, only PHA induced lymphocyte proliferation was suppressed by CS. On day 30, presentation of CS did not have any effect on these immune parameters. These results suggest that the conditioned suppression of the cellular immune function can retain 5-15 days, and extinct after 30 days. 3. CTA was easily induced by one or two CS-UCS parings, and remained robust even after 30 days. These data demonstrate that CIS can be dissociated from CTA, and they may be mediated by different neural mechanisms. 4. Immunohistochemistry assays revealed a broad pattern of c-fos expression throughout the rat brain following the CS-UCS pairing and reexposure to CS, suggesting that many brain regions are involved in CIS. Some brain areas including the solitary tract nucleus (Sol), lateral parabrachial nucleus (LPB) and insular cortex (IC), showed high level c-fos expressions in response to both CS and UCS, suggesting that they may be involved in the transmission and integration of the CS and UCS signals in the brain. There were dense c-FOS positive neurons in the paraverntricular nucleus (PVN) and supraoptic nucleus (SO) of hypothalamus, subfornical organ (SFO) and area postrema (AP) etc. after two trials of CS-UCS paring and after the reexposure to CS 5 days later, but not in the first training and after the extinction of CIS (30 days later). The results reflect that these nuclei may have an important role in CIS expression, and may also response to the immunosuppression of UCS. The conditioned training and reexposure to CS 5 days later induced high level c-fos expression in the cingulate cortex (Cg), central amygdaloid nucleus (Ce), intermediate part of lateral septal nucleus (LSI) and ventrolateral parabrachial nucleus (VLPB) etc. But c-fos induction was not apparent when presenting CS 30 days later. These brain regions are mainly involved in CIS, and may be critical structures in the acquisition and expression of CIS. Some brain regions, including the frontal cortex (Fr), ventral orbital cortex (VO), IC, perirhinal cortex (PRh), LPB and the medial part of solitary nucleus (SolM), showed robust c-FOS expression following the conditioning training and reexposure to CS both on day 5 and day 30, suggesting that they are critically involved in CTA.

Some Extensions of the K-Means Algorithm for Image Segmentation and Pattern Classification

In this paper we present some extensions to the k-means algorithm for vector quantization that permit its efficient use in image segmentation and pattern classification tasks. It is shown that by introducing state variables that correspond to certain statistics of the dynamic behavior of the algorithm, it is possible to find the representative centers fo the lower dimensional maniforlds that define the boundaries between classes, for clouds of multi-dimensional, mult-class data; this permits one, for example, to find class boundaries directly from sparse data (e.g., in image segmentation tasks) or to efficiently place centers for pattern classification (e.g., with local Gaussian classifiers). The same state variables can be used to define algorithms for determining adaptively the optimal number of centers for clouds of data with space-varying density. Some examples of the applicatin of these extensions are also given.

Pattern Motion Perception: Feature Tracking or Integration of Component Motions?

A key question regarding primate visual motion perception is whether the motion of 2D patterns is recovered by tracking distinctive localizable features [Lorenceau and Gorea, 1989; Rubin and Hochstein, 1992] or by integrating ambiguous local motion estimates [Adelson and Movshon, 1982; Wilson and Kim, 1992]. For a two-grating plaid pattern, this translates to either tracking the grating intersections or to appropriately combining the motion estimates for each grating. Since both component and feature information are simultaneously available in any plaid pattern made of contrast defined gratings, it is unclear how to determine which of the two schemes is actually used to recover the plaid"s motion. To address this problem, we have designed a plaid pattern made with subjective, rather than contrast defined, gratings. The distinguishing characteristic of such a plaid pattern is that it contains no contrast defined intersections that may be tracked. We find that notwithstanding the absence of such features, observers can accurately recover the pattern velocity. Additionally we show that the hypothesis of tracking "illusory features" to estimate pattern motion does not stand up to experimental test. These results present direct evidence in support of the idea that calls for the integration of component motions over the one that mandates tracking localized features to recover 2D pattern motion. The localized features, we suggest, are used primarily as providers of grouping information - which component motion signals to integrate and which not to.