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.

Auxetic nuclei in embryonic stem cells exiting pluripotency.

Embryonic stem cells (ESCs) self-renew in a state of naïve pluripotency in which they are competent to generate all somatic cells. It has been hypothesized that, before irreversibly committing, ESCs pass through at least one metastable transition state. This transition would represent a gateway for differentiation and reprogramming of somatic cells. Here, we show that during the transition, the nuclei of ESCs are auxetic: they exhibit a cross-sectional expansion when stretched and a cross-sectional contraction when compressed, and their stiffness increases under compression. We also show that the auxetic phenotype of transition ESC nuclei is driven at least in part by global chromatin decondensation. Through the regulation of molecular turnover in the differentiating nucleus by external forces, auxeticity could be a key element in mechanotransduction. Our findings highlight the importance of nuclear structure in the regulation of differentiation and reprogramming.

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.

STUDY ON THE MODE OF PRIMARY CONTROL IN THE EGG OF GYNOGENETIC CRUCIAN CARP FOR INHIBITING THE DEVELOPMENT OF 2 TYPES OF SPERM NUCLEI

By comparing the different developmental characteristics of two types of sperm nuclei which were from gynogenetic fish (crucian carp) and amphimictic fishes (red carp, red goldfish and sex-reversal red carp) respectively in the eggs of gynogenetic crucian carp, it was preliminarily revealed that there existed selective inhibiting actions of the primary control in the eggs of crucian carp for inhibiting the development of the two types of sperm nuclei. To homologous sperms, the primary control showed weak effect, thus leading to the decondensation of homologous sperm nuclei at different degrees in the eggs of crucian carp. But to heterologous sperms, the primary control showed strong effects, resulting in the total inhibition of the development of heterologous sperm nuclei. Moreover, our experimental results also showed that the different developmental behavior of the two types of sperm nuclei might have a great relationship to the changes of the sex ratio in the population of gynogenetic crucian carp. The infiltration of "the genetic materials in sperm nuclei" into the female nucleus at random might play an important role in male emergence in the naturally gynogenetic population of crucian carp.

Electron spin relaxation by nuclei and holes in single InAs quantum dots

Electron spin relaxation of charged excitons X+ and X2+ are investigated by time-resolved and polarization-resolved photoluminescence spectroscopy. For X+ configuration, the electron spin relaxation shows a typical decay curve induced by hyperfine interaction with nuclei, whereas for X2+ state the electron spin relaxation is affected not only by nuclei but also by electron-hole exchange interaction, leading to a power-law time dependence.

A three-dimensional analytical solution of the microheater temperature before bubble nucleation

A three-dimensional analytical solution of the microheater temperature based on heat diffusion equation is developed and compared with experimental results. Dimensionless parameters are introduced to analyze the temperature rise time and the distribution under steady state. To study the microheater temperatures before bubble nucleation, a set of working fluids and microheaters are considered. It is shown that the dimensionless time xi(-)(0) required for the temperature rise from room to 95% of the steady state temperature is about 75, not dependent on working fluids and microheaters. Heat transfer to the surrounding liquid is mainly caused by conduction, not by convection and radiation mechanisms. The microheater length affects the surface temperature uniformity, while its width influences the steady temperatures significantly, yielding the transition from heterogeneous to homogeneous nucleation mechanism from square microheaters to narrow line microheaters. 

Effect of pulse heating parameters on the microscale bubble dynamics at a microheater surface

We study the effects of pulse heating parameters on the micro bubble behavior of a platinum microheater (100 mu m x 20 mu m) immersed in a methanol pool. The experiment covers the heat fluxes of 10-37 MW/m(2) and pulse frequencies of 25-500 Hz. The boiling incipience is initiated at the superheat limit of methanol, corresponding to the homogeneous nucleation. Three types of micro boiling patterns are identified. The first type is named as the bubble explosion and regrowth, consisting of a violent explosive boiling and shrinking, followed by a slower bubble regrowth and subsequent shrinking, occurring at lower heat fluxes. The second type, named as the bubble breakup and attraction, consists of the violent explosive boiling, bubble breakup and emission, bubble attraction and coalescence process, occurring at higher heat fluxes than those of the first type. The third type, named as the bubble size oscillation and large bubble formation, involves the initial explosive boiling, followed by a short periodic bubble growth and shrinking. Then the bubble continues to increase its size, until a constant bubble size is reached which is larger than the microheater length. 

Periodic bubble emission and appearance of an ordered bubble sequence (train) during condensation in a single microchannel

Condensation of steam in a single microchannel, silicon test section was investigated visually at low flow rates. The microchannel was rectangular in cross-section with a depth of 30 pm, a width of 800 mu m and a length of 5.0 mm, covered with a Pyrex glass to allow for visualization of the bubble formation process. By varying the cooling rate during condensation of the saturated water vapor, it was possible to control the shape, size and frequency of the bubbles formed. At low cooling rates using only natural air convection from the ambient environment, the flow pattern in the microchannel consisted of a nearly stable elongated bubble attached upstream (near the inlet) that pinched off into a train of elliptical bubbles downstream of the elongated bubble. It was observed that these elliptical bubbles were emitted periodically from the tip of the elongated bubble at a high frequency, with smaller size than the channel width. The shape of the emitted bubbles underwent modifications shortly after their generation until finally becoming a stable vertical ellipse, maintaining its shape and size as it flowed downstream at a constant speed. These periodically emitted elliptical bubbles thus formed an ordered bubble sequence (train). At higher cooling rates using chilled water in a copper heat sink attached to the test section, the bubble formation frequency increased significantly while the bubble size decreased, all the while forming a perfect bubble train flowing downstream of the microchannel. The emitted bubbles in this case immediately formed into a circular shape without any further modification after their separation from the elongated bubble upstream. The present study suggests that a method for controlling the size and generation frequency of microbubbles could be so developed, which may be of interest for microfluidic applications. The breakup of the elongated bubble is caused by the large Weber number at the tip of the elongated bubble induced by the maximum vapor velocity at the centerline of the microchannel inside the elongated bubble and the smaller surface tension force of water at the tip of the elongated bubble.

Transient flow patterns and bubble slug lengths in parallel microchannels with oxygen gas bubbles produced by catalytic chemical reactions

Transient flow patterns and bubble slug lengths were investigated with oxygen gas (O-2) bubbles produced by catalytic chemical reactions using a high speed camera bonded with a microscope. The microreactor consists of an inlet liquid plenum, nine parallel rectangular microchannels followed by a micronozzle, using the MEMS fabrication technique. The etched surface was deposited by the thin platinum film, which is acted as the catalyst. Experiments were performed with the inlet mass concentration of the hydrogen peroxide from 50% to 90% and the pressure drop across the silicon chip from 2.5 to 20.0 kPa. The silicon chip is directly exposed in the environment thus the heat released via the catalytic chemical reactions is dissipated into the environment and the experiment was performed at the room temperature level. It is found that the two-phase flow with the catalytic chemical reactions display the cyclic behavior. A full cycle consists of a short fresh liquid refilling stage, a liquid decomposition stage followed by the bubble slug flow stage. At the beginning of the bubble slug flow stage, the liquid slug number reaches maximum, while at the end of the bubble slug flow stage the liquid slugs are quickly flushed out of the microchannels. Two or three large bubbles are observed in the inlet liquid plenum, affecting the two-phase distributions in microchannels. The bubble slug lengths, cycle periods as well as the mass flow rates are analyzed with different mass concentrations of hydrogen peroxide and pressure drops. The bubble slug length is helpful for the selection of the future microreactor length ensuring the complete hydrogen peroxide decomposition. Future studies on the temperature effect on the transient two-phase flow with chemical reactions are recommended.

Study on the kinetics of hydrate formation in a bubble column

Gas hydrate formation experiments were performed using methane in the presence of tetrahydrofuran (THF) in aqueous solution in a transparent bubble column in which a single pipe or a sintered plate was used to produce bubbles. The mole fraction of THF in aqueous solution was fixed at 6%. The hydrate formation kinetic behaviors on the surface of the rising bubble, the mechanical stability of hydrate shell formed on the surface of the bubble, the interactions among the bubbles with hydrate shell were observed and investigated morphologically. The rise velocities of individual bubbles with hydrate shells of different thickness and the consumption rates of methane gas were measured. A kinetic model was developed to correlate the experimentally measured gas consumption rate data. It was found that the hydrate formation rate on the surface of the moving bubble was high, but the formed hydrate shell was not very easy to be broken up. The bubbles with hydrate shells tended to agglomerate rather than merge into bigger bubble. This kind of characteristic of hydrate shell hindered the further formation of hydrate and led to the lower consumption rate of methane. The consumption rate of methane was found to increase with the decrease of temperature or increase of pressure. The increase of gas flux led to a linear increase in consumption rate of methane. It was demonstrated that the developed kinetic model could be used to correlate the consumption rate satisfyingly.