A serine proteinase inhibitor from frog eggs with bacteriostatic activity

By Sephadex G-50 gel filtration, Resource Q anionic exchange and C4 reversed phase liquid high performance liquid chromatography, a proteinase inhibitor protein (Ranaserpin) was identified and purified from the eggs of the odour frog, Rana grahami. The protein displayed a single band adjacent to the molecular weight marker of 14.4 kDa analyzed by SDS-PAGE. The inhibitor protein homogeneity and its molecular weight were confirmed again by MALDI-TOF mass spectrometry analysis. The MALDI-TOF mass spectrum analysis gave this inhibitor protein an m/z of 14422.26 that was matched well with the result from SDS-PAGE. This protein is a serine proteinase inhibitor targeting multiple proteinases including trypsin, elastase, and subtilisin. Ranaserpin inhibited the proteolytic activities of trypsin, elastase, and subtilisin. It has an inhibitory constant (K-i) of 6.2 x 10(-8) M, 2.7 x 10(-7) M and 2.2 x 10(-8) M for trypsin, elastase, and subtilisin, respectively. This serine proteinase inhibitor exhibited bacteriostatic effect on Gram-positive bacteria Bacillus subtilis (ATCC 6633). It was suggested that ranaserpin might act as a defensive role in resistance to invasion of pests or pathogens. This is the first report of serine proteinase inhibitor and its direct defensive role from amphibian eggs. (C) 2007 Elsevier Inc. All rights reserved.

Increasing the genetic uniformity of bighead carp [Aristichthys nobilis (Richardson)] by means of spontaneous diploidization of gynogenetically activated eggs

Three groups of gynogenetic diploid bighead carp were successfully obtained by means of artificial gynogenesis. The activation rates of gynogenesis varied from 75.9% to 98.8%, and the frequency of spontaneous diploidization was around 0.4%. Over 2000 normally gynogenetic diploid fry were obtained in three gynogenetic groups. The haploid karyotype consisted of nine metacentric, 12 submetacentric, three subtelocentric chromosomes and 45 arms. The chromosome number was 48 from gynogenetic diploid. The results showed that the genetic material of offspring was maternal. The aneuploid hybrid embryos of bighead carp and Xingguo red common carp with chromosome numbers ranging from 28 to 73 did not survive post hatch, likely the result of incompatibility between the nucleus and the cytoplasm of two parents. Sixty RAPD primers from three groups were used for total DNA amplification of gynogenetic offspring, maternal and 'paternal' fish. A total of 451 bands were amplified from three kinds of samples above. From maternal bighead carp, 256 bands were amplified; however, there were 251 shared bands between maternal and gynogenetic bighead carp. From artificial gynogenetic offspring, two 'paternal' DNA segments without an expression function were found. An UPGMA tree showed that gynogenetic offspring were closely clustered and the genetic identity among them was very high (0.956).

Oxidative damage in unfertilized eggs of Chinese rare minnow (Gobiocypris rarus) exposed to nonylphenol

In the present study, female Chinese rare minnows (Gobiocypris rarus) were used as in vivo models and exposed to nonylphenol (NP) at concentrations of 1 to 200 mu g/L for 21 d under semistatic conditions. Molecular biomarkers of oxidative stress were measured in unfertilized eggs and included reactive oxygen species (ROS), lipid peroxidation products (thiobarbituric acid-reactive substances [TBARS] and protein carbonyl), superoxide dismutase activity, and glutathione. Cathepsin D activity as an indicator of egg viability also was assayed. Nonylphenol induced ROS formation in unfertilized eggs in all exposed groups compared to the controls. The levels of protein carbonyl and TBARS in unfertilized eggs were significantly increased (p < 0.05) at 10 to 200 and 100 to 200 mu g/L, respectively. Good positive correlations were shown between ROS induction and levels of TBARS and protein carbonyl in eggs (R = 0.918, p < 0.05 and R = 0.784, p < 0.05, respectively). Superoxide dismutase activity in eggs was significantly inhibited (p < 0.05) in the 50 to 200 mu g/L exposure groups. Glutathione levels in eggs were significantly depleted (p < 0.05) at 100 to 200 mu g/L concentrations. In addition, ROS induction resulted in oxidative damage to lipid and protein in chorions. Significant reductions (p < 0.05) of the protein and lipid contents in chorions were both found in the 50 to 200 mu g/L exposure groups. A previous study found that NP exposure could lead to chorion thinning in zebra fish. Thus, the reductions in protein and lipid contents in chorion could be the reason for chorion thinning by NP exposure. Meanwhile, cathepsin D activity was significantly inhibited (p < 0.05) in all exposure groups. The results demonstrated that NP-induced oxidative stress could damage the chorion of unfertilized eggs and lead to a decline in gamete quality in female Chinese rare minnow.

Identification of differentially expressed genes from the cross-subfamily cloned embryos derived from zebrafish nuclei and rare minnow enucleated eggs

Cross-species nuclear transfer (NT) has been used to retain the genetic viability of a species near extinction. However, unlike intra-species NT, most embryos produced by cross-species NT were unable to develop to later stages due to incompatible nucleocytoplasmic interactions between the donor nuclei and the recipient cytoplasm from different species. To study the early nucleocytoplasmic interaction in cross-species NT, two laboratory fish species (zebrafish and rare minnow) from different subfamilies were used to generate cross-subfamily NT embryos in the present study. Suppression subtractive hybridization (SSH) was performed to screen out differentially expressed genes from the forward and reverse subtractive cDNA libraries. After dot blot and real-time PCR analysis, 80 of 500 randomly selective sequences were proven to be differentially expressed in the cloned embryos. Among them, 45 sequences shared high homology with 28 zebrafish known genes, and 35 sequences were corresponding to 22 novel expressed sequence tags (ESTs). Based on functional clustering and literature mining analysis, up-and down-regulated genes in the cross-subfamily cloned embryos were mostly relevant to transcription and translation initiation, cell cycle regulation, protein binding, etc. To our knowledge, this is the first report on the determination of genes involved in the early development of cross-species NT embryos of fish. (C) 2007 Elsevier Inc. All rights reserved.

Cytoplasmic impact on cross-genus cloned fish derived from transgenic common carp (Cyprinus carpio) nuclei and goldfish (Carassius auratus) enucleated eggs

In previous studies of nuclear transplantation, most cloned animals were obtained by intraspecies nuclear transfer and are phenotypically identical to their nuclear donors; furthermore, there was no further report on successful fish cloning since the report of cloned zebrafish. Here we report the production of seven cross-genus cloned fish by transferring nuclei from transgenic common carp into enucleated eggs of goldfish. Nuclear genomes of the cloned fish were exclusively derived from the nuclear donor species, common carp, whereas the mitochondrial DNA from the donor carp gradually disappeared during the development of nuclear transfer (NT) embryos. The somite development process and somite number of nuclear transplants were consistent with the recipient species, goldfish, rather than the nuclear donor species, common carp. This resulted in a long-lasting effect on the vertebral numbers of the cloned fish, which belonged to the range of goldfish. These demonstrate that fish egg cytoplasm not only can support the development driven by transplanted nuclei from a distantly related species at the genus scale but also can modulate development of the nuclear transplants.

Effect of temperature and photoperiod on hatching of eggs of Tokunagayusurika akamusi (Tokunaga) (Diptera : Chironomidae)

The hatching time of eggs of Tokunagayusurika akamusi (Tokunaga) decreased significantly when temperature was increased from 5degreesC to 25degreesC; eggs fail to hatch at 30degreesC. The percentage of T akamusi eggs that developed into normal larva also was negatively correlated with temperature. The hatching time decreased with lengthened photoperiod.

High-frequency characterization of packaging network in to-can photodiode modules

A simple method for analyzing the effects of TO packaging network on the high-frequency response of photodiode modules is presented. This method is established based on the relations of the scattering parameters of the packaging network, photodiode chip, and module. It is shown that the results obtained by this method agree well with those obtained by the conventional comparison method. The proposed method is much more convenient since only the electrical domain measurements are required. (C) 2008 Wiley Periodicals, Inc.

Frequency bandwidth estimation of TO packaging techniques for laser modules

Two simple methods for estimating the potential modulation bandwidth of TO packaging technique are presented. The first method is based upon the comparison of the measured frequency responses of the laser diodes and the TO laser modules, and the second is from the equivalent circuit for the test fixture, the TO header, the submount and the bonding wire. It is shown that the TO packaging techniques used in the experiments can potentially achieve a frequency bandwidth of over 10.5 GHz, and the two proposed methods give similar results.

Methods for characterization of packaging parasitics of semiconductor lasers

Two novel methods for analyzing the parasitics of packaging networks are proposed based on the relations between the scattering parameters of a semiconductor laser before and after packaging, and the experiments are designed and performed using our methods. It is found that the analysis results of the two methods are in good agreement with the measurements. Either of the two methods can provide an alternative approach for characterizing the packaging parasitics for semiconductor lasers, and both are convenient due to the developed measurement techniques. (c) 2005 Wiley Periodicals, Inc.

Silicon bench for passive coupling and packaging of high-frequency optoelectronic devices

Coupling and packaging have become decisive factors in the final performance and cost of high-frequency optoelectronic devices. Here, we report the design and successful fabrication of a silicon bench that integrates a V-groove and high-frequency coplanar waveguide (CPW) on the same high-resistivity silicon wafer as an effective optoelectronic packaging solution.

Potential frequency bandwidth estimation of TO packaging techniques for photodiode modules

Scattering parameters of photodiode chip, TO header and TO packaged module are measured, and the effects of TO packaging network on the high-frequency response of photodiode are investigated. Based on the analysis, the potential bandwidth of TO packaging techniques is estimated from the scattering parameters of the TO packaging network. Another method for estimating the potential bandwidth from the equivalent circuit for the TO packaged photodiode model is also presented. The results obtained using both methods show that the TO packaging techniques used in the experiments can potentially achieve a frequency bandwidth of 22 GHz.

Bonding-wire compensation effect on the packaging parasitics of optoelectronic devices

The effect of bonding-wire compensation on the capacitances of both the submount and the laser diode is demonstrated in this paper. The measured results show that the small-signal magnitude-frequency responses of the TO packaged laser and photodiode modules can be improved by properly choosing the length of the bonding wire. After packaging, the phase-frequency responses of the laser modules can also be significantly improved (c) 2005 Wiley Periodicals, Inc.

Microwave packaging for 10 Gb/s EML modulators - art. no. 60201O

A novel microwave packaging technique for 10Gb/s electro-absorption modulator integrated with distributed feedback laser (EML) is presented. The packaging parasitics and intrinsic parasitics are both well considered, and the packaging circuit was synthetically designed to compensate for the intrinsic parasitic of the chip. A butterfly-packaged EMI module has been successfully developed to prove that. The small-signal modulation bandwidth of the butterfly-packaged module is about 10 GHz. Optical fiber transmission experiments have shown that the module can be used for 10Gb/s optical transmission system. After transmission through 40km,. the power penalty is less than 1 dBm at a bit-error-rate of 10-12.

Design and Fabrication of Low-microwave loss Coplannar Waveguide and precise V groove on Silicon Substrate for Optoelectronic Packaging - art. no. 601938

Optoelectronic packaging has become a most important factor that influences the final performance and cost of the module. In this paper, low microwave loss coplanar waveguide(CPW) on high resistivity silicon(HRS) and precise V groove in silicon substrate were successfully fabricated. The microwave attenuation of the CPW made on HRS with the simple process is lower than 2 dB/cm in the frequency range of 0 similar to 26GHz, and V groove has the accuracy in micro level and smooth surface. These two techniques built a good foundation for high frequency packaging and passive coupling of the optoelectronic devices. Based on these two techniques, a simple high resistivity silicon substrate that integrated V groove and CPW for flip-chip packaging of lasers was completed. It set a good example for more complicate optoelectronic packaging.