Parallel and Convergent Evolution of the Dim-Light Vision Gene RH1 in Bats (Order: Chiroptera)

Rhodopsin, encoded by the gene Rhodopsin (RH1), is extremely sensitive to light, and is responsible for dim-light vision. Bats are nocturnal mammals that inhabit poor light environments. Megabats (Old-World fruit bats) generally have well-developed eyes, while microbats (insectivorous bats) have developed echolocation and in general their eyes were degraded, however, dramatic differences in the eyes, and their reliance on vision, exist in this group. In this study, we examined the rod opsin gene (RH1), and compared its evolution to that of two cone opsin genes (SWS1 and M/LWS). While phylogenetic reconstruction with the cone opsin genes SWS1 and M/LWS generated a species tree in accord with expectations, the RH1 gene tree united Pteropodidae (Old-World fruit bats) and Yangochiroptera, with very high bootstrap values, suggesting the possibility of convergent evolution. The hypothesis of convergent evolution was further supported when nonsynonymous sites or amino acid sequences were used to construct phylogenies. Reconstructed RH1 sequences at internal nodes of the bat species phylogeny showed that: (1) Old-World fruit bats share an amino acid change (S270G) with the tomb bat; (2) Miniopterus share two amino acid changes (V104I, M183L) with Rhinolophoidea; (3) the amino acid replacement I123V occurred independently on four branches, and the replacements L99M, L266V and I286V occurred each on two branches. The multiple parallel amino acid replacements that occurred in the evolution of bat RH1 suggest the possibility of multiple convergences of their ecological specialization (i.e., various photic environments) during adaptation for the nocturnal lifestyle, and suggest that further attention is needed on the study of the ecology and behavior of bats.

Distinct Evolutionary Patterns Between Two Duplicated Color Vision Genes Within Cyprinid Fishes

We investigated the molecular evolution of duplicated color vision genes (LWS-1 and SWS2) within cyprinid fish, focusing on the most cavefish-rich genus-Sinocyclocheilus. Maximum likelihood-based codon substitution approaches were used to analyze the evolution of vision genes. We found that the duplicated color vision genes had unequal evolutionary rates, which may lead to a related function divergence. Divergence of LWS-1 was strongly influenced by positive selection causing an accelerated rate of substitution in the proportion of pocket-forming residues. The SWS2 pigment experienced divergent selection between lineages, and no positively selected site was found. A duplicate copy of LWS-1 of some cyprinine species had become a pseudogene, but all SWS2 sequences remained intact in the regions examined in the cyprinid fishes examined in this study. The pseudogenization events did not occur randomly in the two copies of LWS-1 within Sinocyclocheilus species. Some cave species of Sinocyclocheilus with numerous morphological specializations that seem to be highly adapted for caves, retain both intact copies of color vision genes in their genome. We found some novel amino acid substitutions at key sites, which might represent interesting target sites for future mutagenesis experiments. Our data add to the increasing evidence that duplicate genes experience lower selective constraints and in some cases positive selection following gene duplication. Some of these observations are unexpected and may provide insights into the effect of caves on the evolution of color vision genes in fishes.

Identification of a WSSV neutralizing scFv antibody by phage display technology and in vitro screening

White spot syndrome virus (WSSV) is one of the most significant viral pathogens causing high mortality and economic damage in shrimp aquaculture. Although intensive efforts were undertaken to detect and characterize WSSV infection in shrimp during the last decade, we still lack methods either to prevent or cure white spot disease. Most of the studies on neutralizing antibodies from sera have been performed using in vivo assays. For the first time, we report use of an in vitro screening method to obtain a neutralizing scFv antibody against WSSV from a previously constructed anti-WSSV single chain fragment variable region (scFv) antibody phage display library. From clones that were positive for WSSV by ELISA, 1 neutralizing scFv antibody was identified using an in vitro screening method based on shrimp primary lymphoid cell cultures. The availability of a neutralizing antibody against the virus should accelerate identification of infection-related genes and the host cell receptor, and may also enable new approaches to the prevention and cure of white spot disease.

A screening study on persistent bioaccumulation toxins (PBT) in fuel combustion soot

Persistent bioaccumulative toxins (PBTs) are organic substances that are persistent, bioaccumulative and can cause severe toxic effects (e.g. potential oncogens, mutagenic, endocrine disrupters) to human health or environment which are the ones that need special attention. PBT chemicals could be released to the environment from several types of sources and are ubiquitous in environment. However, fast and efficiency monitoring and assessment methods to investigate PBTs in environment are still lacking. In this study, a cleaning-up procedure of analyzing PBTs in fuels combustion soot was developed and its performance was assessed through comparing the chromatograms of crude extracts with their cleaned extracts after the cleaning-up procedure. The results showed that polycyclic aromatic compounds (PACs) were the main components in fuel combustion soot and the clean-up procedure developed in this paper can be well used as the method of analyzing PBTs in fuels combustion soot.

Characterization of two membrane-associated protease genes obtained from screening out-membrane protein genes of Flavobacterium columnare G(4)

In order to identify genes encoding the outer membrane proteins (OMPs) of the myxobacter Flavobacterium columnare G(4), the expression library of the bacterium was screened by using rabbit antisera developed against its OMPs. Positive colonies of Escherichia coli M15 containing fragments encoding the bacterial OMPs were selected for cloning the relevant genes by genomic walking methods. Two genes encoding a membrane-associated zinc metalloprotease and prolyl oligopeptidase are reported in this paper. The membrane-associated zinc metalloprotease gene (map) is 1800 bp in length, coding for 449 amino acids (aa). Despite the presence of a conserved motif HEXXH for all metalloproteases, the special HEXXH similar to 32 aa similar to E motif of the F. columnare G(4) Map and its low level of identity with other reported zinc-containing metalloproteases may imply that the membrane-associated zinc metalloprotease of F. columnare G(4) represents a new family of zincins. The gene encoding prolyl oligopeptidase (Pop), a serine proteinase, is 2352 bp in length, coding for 649 aa. Sequence homology analysis revealed that the Pop is also novel as it has <50% identity with other reported prolyl oligopeptidase family proteins. The present study represents the first to employ anti-fish bacterial OMP sera to screen genes of membrane-associated proteases of fish pathogenic bacteria, and to provide necessary information for the examination of the role of the two genes in the infection and pathogenesis of F. columnare.

Differential screening and characterization analysis of the egg envelope glycoprotein ZP3 cDNAs between gynogenetic and gonochoristic crucian carp

Gynogenetic silver crucian carp, Carassius auratus gibelio, is an intriguing model. system. In the present work, a systemic study has been initiated by introducing suppression subtractive hybridization technique into this model system to identify the differentially expressed genes in oocytes between gynogenetic silver crucian carp and its closely related gonochoristic color crucian carp. Five differential cDNA fragments were identified from the preliminary screening, and two of them are ZP3 homologues. Moreover, the full length ZP3 cDNAs were cloned from their oocyte cDNA libraries. The length of ZP3 cDNAs were 1378 bp for gyno-carp and 1367 bp for gono-carp, and they can be translated into proteins with 435 amino acids. Obvious differences are not only in the composition of amino acids, but also in the number of potential O-linked oligosaccharide sites. In addition, gyno-carp ZP3 amino acid sequence has an unexpected higher identity value with common carp (83.5%) than that with the closely related gono-carp (74.7%). The unique homology may be originated from the ancient hybridization. Northern blot analysis confirmed that expression of the ZP3 gene occurred exclusively in the oocytes. Because O-linked oligosaccharides on ZP3 have been demonstrated to play very important roles in fertilization, it is suggested that the extra O-linked glycosylation sites may be related to the unique sperm-egg recognition mechanism in gynogenesis.

A new enzyme immunoassay for PCDD/F TEQ screening in environmental samples: Comparison to micro-EROD assay and to chemical analysis

A new Enzyme ImmunoAssay (EIA) for PCDD/F TEQ measurement in extracts of environmental samples was described. The bioassay TEQ which derived from EIA and EROD were compared with each other and with results from chemical analysis. For all environmental samples, the EROD-TEQ is higher than the value from chemical analysis. However, the EIA-TEQ is much more identical with the value from chemical analysis. Our results indicate that the EIA assay is a complementary method to the EROD assay and should be useful as a rapid and sensitive screening tool for environmental samples in many situations. (C) 1999 Elsevier Science Ltd. All rights reserved

Theoretical analysis and experimental study of the space-charge-screening effect in uni-traveling-carrier photodiode

A new optimized structure of an UTC (uni-traveling-carrier) photodiode is developed and epitaxied by metal-organic chemical vapor deposition. We fabricated a UTC photodiode of 30 mu m in diameter. Theoretical simulation based on drift-diffusion model was used to analyze the space-charge-screening effect in UTC photodiode primarily in two aspects: the carrier concentrations and the space electric field. The simulation results were generally in agreement with the experimental data.

A programmable SIMD vision chip for real-time vision applications

A programmable vision chip for real-time vision applications is presented. The chip architecture is a combination of a SIMD processing element array and row-parallel processors, which can perform pixel-parallel and row-parallel operations at high speed. It implements the mathematical morphology method to carry out low-level and mid-level image processing and sends out image features for high-level image processing without I/O bottleneck. The chip can perform many algorithms through software control. The simulated maximum frequency of the vision chip is 300 MHz with 16 x 16 pixels resolution. It achieves the rate of 1000 frames per second in real-time vision. A prototype chip with a 16 x 16 PE array is fabricated by the 0.18 mu m standard CMOS process. It has a pixel size of 30 mu m x 40 mu m and 8.72 mW power consumption with a 1.8 V power supply. Experiments including the mathematical morphology method and target tracking application demonstrated that the chip is fully functional and can be applied in real-time vision applications.

A 1,000 Frames/s Programmable Vision Chip with Variable Resolution and Row-Pixel-Mixed Parallel Image Processors

A programmable vision chip with variable resolution and row-pixel-mixed parallel image processors is presented. The chip consists of a CMOS sensor array, with row-parallel 6-bit Algorithmic ADCs, row-parallel gray-scale image processors, pixel-parallel SIMD Processing Element (PE) array, and instruction controller. The resolution of the image in the chip is variable: high resolution for a focused area and low resolution for general view. It implements gray-scale and binary mathematical morphology algorithms in series to carry out low-level and mid-level image processing and sends out features of the image for various applications. It can perform image processing at over 1,000 frames/s (fps). A prototype chip with 64 x 64 pixels resolution and 6-bit gray-scale image is fabricated in 0.18 mu m Standard CMOS process. The area size of chip is 1.5 mm x 3.5 mm. Each pixel size is 9.5 mu m x 9.5 mu m and each processing element size is 23 mu m x 29 mu m. The experiment results demonstrate that the chip can perform low-level and mid-level image processing and it can be applied in the real-time vision applications, such as high speed target tracking.

A novel vision chip for high-speed target tracking

This paper presents a novel vision chip for high-speed target tracking. Two concise algorithms for high-speed target tracking are developed. The algorithms include some basic operations that can be used to process the real-time image information during target tracking. The vision chip is implemented that is based on the algorithms and a row-parallel architecture. A prototype chip has 64 x 64 pixels is fabricated by 0.35 pm complementary metal-oxide-semiconductor transistor (CMOS) process with 4.5 x 2.5 mm(2) area. It operates at a rate of 1000 frames per second with 10 MHz chip main clock. The experiment results demonstrate that a high-speed target can be tracked in complex static background and a high-speed target among other high-speed objects can be tracked in clean background.

The research of range-gated laser night vision system on cars - art. no. 68351V

The characteristic of several night imaging and display technologies on cars are introduced. Compared with the current night vision technologies on cars, Range-gated technology can eliminate backscattered light and increase the SNR of system. The theory of range-gated image technology is described. The plan of range-gated system on cars is designed; the divergence angle of laser can be designed to change automatically, this allows overfilling of the camera field of view to effectively attenuate the laser when necessary. Safety range of the driver is calculated according to the theory analysis. Observation distance of the designed system is about 500m which is satisfied with the need of safety driver range.

Research of Face Location System Based on Human Vision Simulations

In this paper we present a robust face location system based on human vision simulations to automatically locate faces in color static images. Our method is divided into four stages. In the first stage we use a gauss low-pass filter to remove the fine information of images, which is useless in the initial stage of human vision. During the second and the third stages, our technique approximately detects the image regions, which may contain faces. During the fourth stage, the existence of faces in the selected regions is verified. Having combined the advantages of Bottom-Up Feature Based Methods and Appearance-Based Methods, our algorithm performs well in various images, including those with highly complex backgrounds.