An Effective Algorithm to Find a Cost Minimizing Gateway Deployment for Node-Replaceable Wireless Sensor Networks.

In this paper, we present an efficient way to find a gateway deployment for a given sensor network topology. We assume that the expired sensors and gateways can be replaced and the locations of the gateways are chosen among the given sensor nodes. The objective is to find a gateway deployment that minimizes the cost per unit time, which consists of the maintenance and installation costs. The proposed algorithm creates a cost reference and uses it to find the optimal deployment via a divide and conquer algorithm. Comparing all cases is the most reliable way to find the optimal gateway deployment, but this is practically impossible to calculate, since its computation time increases exponentially as the number of nodes increases. The method we propose increases linearly, and so is suitable for large scale networks. Additionally, compared to stochastic algorithms such as the genetic algorithm, this methodology has advantages in computational speed and accuracy for a large number of nodes. We also verify our methodology through several numerical experiments.

Preparation and Characterization of Carbon Fibers from Lyocell Precursors Grafted with Polyacrylamide via Electron-Beam Irradiation.

Carbon fibers, which act as reinforcements in many applications, are often obtained from polyacrylonitrile (PAN). However, their production is expensive and results in waste problems. Therefore, we focused on producing carbon fibers from lyocell, a cellulose-based material, and analyzed the effects of the process parameters on their mechanical properties and carbon yields. Lyocell was initially grafted with polyacrylamide (PAM) via electron-beam irradiation (EBI) and was subsequently stabilized and carbonized. Thermal analysis showed that PAM grafting increased the carbon yields to 20% at 1000 °C when compared to that of raw lyocell, which degraded completely at about 600 °C. Stabilization further increased this yield to 55%. The morphology of the produced carbon fibers was highly dependent on PAM concentration, with fibers obtained at concentrations ≤0.5 wt.% exhibiting clear, rigid, and round cross-sections with smooth surfaces, whereas fibers obtained from 2 and 4 wt.% showed peeling surfaces and attachment between individual fibers due to high viscosity of PAM. These features affected the mechanical properties of the fibers. In this study, carbon fibers of the highest tensile strength (1.39 GPa) were produced with 0.5 wt.% PAM, thereby establishing the feasibility of using EBI-induced PAM grafting on lyocell fabrics to produce high-performance carbon fibers with good yields.

A Discrete Velocity Kinetic Model with Food Metric: Chemotaxis Traveling Waves.

We introduce a mesoscopic scale chemotaxis model for traveling wave phenomena which is induced by food metric. The organisms of this simplified kinetic model have two discrete velocity modes, [Formula: see text] and a constant tumbling rate. The main feature of the model is that the speed of organisms is constant [Formula: see text] with respect to the food metric, not the Euclidean metric. The uniqueness and the existence of the traveling wave solution of the model are obtained. Unlike the classical logarithmic model case there exist traveling waves under super-linear consumption rates and infinite population pulse-type traveling waves are obtained. Numerical simulations are also provided.

Cryopreservation of putative pre-pubertal bovine spermatogonial stem cells by slow freezing.

Development of techniques for the preservation of mammalian spermatogonial stem cells (SSCs) is a critical step in commercial application of SSC based technologies, including species preservation, amplification of agriculturally valuable germ lines, and human fertility preservations. The objective of this study was to develop an efficient cryopreservation protocol for preservation of bovine SSCs using a slow freezing technique. To maximize the efficiency of SSC cryopreservation, the effects of various methods (tissue vs. cell freezing) and cryoprotective agents (trehalose, sucrose, and polyethylene glycol [PEG]) were tested. Following thawing, cells were enriched for undifferentiated spermatogonia by differential plating and evaluated for recovery rate, proliferation capacity, and apoptosis. Additionally, putative stem cell activity was assessed using SSC xenotransplantation. The recovery rate, and proliferation capacity of undifferentiated spermatogonia were significantly greater for germ cells frozen using tissue freezing methods compared to cell freezing methods. Cryopreservation in the presence of 200 mM trehalose resulted in significantly greater recovery rate, proliferation capacity, and apoptosis of germ cells compared to control. Furthermore, cryopreservation using the tissue freezing method in the presence of 200 mM trehalose resulted in the production of colonies of donor-derived germ cells after xenotransplantation into recipient mouse testes, indicating putative stem cell function. Collectively, these data indicate that cryopreservation using tissue freezing methods in the presence of 200 mM trehalose is an efficient cryopreservation protocol for bovine SSCs.

Rigid Formation Control on a Sphere: A Heterogeneous System Approach.

We consider a heterogeneous multiagent system for tracking multiple targets with a rigid formation on a unit sphere, where the targets and chasing agents are governed by single-and double-integrator models, respectively. To make asymptotic rendezvous between agents and their corresponding targets, we use an autonomous system consisting of attraction forces and velocity alignments. If the target's position, velocity, and acceleration information are available, we derive a multiagent system for complete rendezvous and obtain its exponential convergence result. If we have only the location and velocity information of the targets, we provide an autonomous system for practical rendezvous and the corresponding mathematical analysis. To prove the main results, we employ frame-rotation-structure decomposition for the double-integrator model and the geometric properties of a rigid formation on a sphere. We also provide numerical simulations to confirm our mathematical results and apply them to multiagent dynamics with a rigid formation that patrols the boundary line for a certain area on the sphere.

Target Tracking Systems on a Sphere With Topographic Information.

We consider the target tracking problem on a sphere with topographic structure. For a given moving target on the unit sphere, we suggest a double-integrator autonomous system of multiple agents that track the given target under the topographic influence. Through this dynamic system, we can obtain a control design for target tracking on the sphere and the adapted topographic data provides an efficient agent trajectory. The topographic information, described as a form of friction in the double-integrator system, affects the velocity and acceleration of the target and agents. The target information required by the tracking agents consists of position, velocity, and acceleration. We can obtain practical rendezvous results when agents utilize only target position and velocity information. If the acceleration data of the target is accessible, we can get the complete rendezvous result using an additional control term in the form of the Coriolis force. We provide mathematically rigorous proofs for these results and present numerical experiments that can be visually confirmed.

Co-synthesis and Electrochemical Investigation of the Nitrogen-Doped Carbon Layer with Metallic Nano Beads on the SiOx Anode for Lithium Secondary Batteries.

The high theoretical capacity (∼2000 mAh g-1) of silicon suboxide (SiOx, with 1 < x < 2) can solve the energy density issue of the graphite anode in Li-ion batteries. In addition, it has an advantage in terms of volume expansion or side reactions compared to pure Si or Li metals, which are considered as next-generation anode materials. However, the loading content of SiOx is limited in commercial anodes because of its low cycle stability and initial coulombic efficiency. In this study, a nitrogen-doped carbon layer with Cu beads (N-C/Cu) derived from copper phthalocyanine (CuPc) is applied to a SiOx electrode to improve its electrochemical performance. The SiOx electrode is simultaneously coated with a Cu- and N-doped carbon layer using CuPc. N-C/Cu synergistically enhances the electric conductivity of the electrode, thus improving its electrochemical performance. The SiOx/N-C/Cu composite has better cyclability and higher capacity (1095.5 mAh g-1) than the uncoated electrode, even after 200 cycles in the 0.5 C condition. In full-cell cycling with NCM811 cathodes, the SiOx (60 wt % of SiOx, with a n/p ratio of 1.1) and graphite-mixed (7.8 wt % of SiOx, with a n/p ratio of 1.1) anodes also show improved electrochemical performances in the same conditions.

Development of cyclic peptomer inhibitors targeting the polo-box domain of polo-like kinase 1.

The polo-box domain (PBD) of polo-like kinase 1 (Plk1) is essentially required for the function of Plk1 in cell proliferation. The availability of the phosphopeptide-binding pocket on PBD provides a unique opportunity to develop novel protein-protein interaction inhibitors. Recent identification of a minimal 5-residue-long phosphopeptide, PLHSpT, as a Plk1 PBD-specific ligand has led to the development of several peptide-based inhibitors, but none of them is cyclic peptide. Through the combination of single-peptoid mimics and thio-ether bridged cyclization, we successfully demonstrated for the first time two cyclic peptomers, PL-116 and PL-120, dramatically improved the binding affinity without losing mono-specificity against Plk1 PBD in comparison with the linear parental peptide, PLHSpT. These cyclic peptomers could serve as promising templates for future drug designs to inhibit Plk1 PBD.

Integrated dataset of the Korean Genome and Epidemiology Study cohort with estimated air pollution data.

Public concern about the adverse health effects of air pollution has grown rapidly in Korea, and there has been increasing demand for research on ways to minimize the health effects of air pollution. Integrating large epidemiological data and air pollution exposure levels can provide a data infrastructure for studying ambient air pollution and its health effects. The Korean Genome and Epidemiology Study (KoGES), a large population-based study, has been used in many epidemiological studies of chronic diseases. Therefore, KoGES cohort data were linked to air pollution data as a national resource for air pollution studies. Air pollution data were produced using community multiscale air quality modeling with additional adjustment of monitoring data, satellite-derived aerosol optical depth, normalized difference vegetation index, and meteorological data to increase the accuracy and spatial resolution. The modeled air pollution data were linked to the KoGES cohort based on participants' geocoded residential addresses in grids of 1 km (particulate matter) or 9 km (gaseous air pollutants and meteorological variables). As the integrated data become available to all researchers, this resource is expected to serve as a useful infrastructure for research on the health effects of air pollution.

Synergistic effects of glutathione and ß-mercaptoethanol treatment during in vitro maturation of porcine oocytes on early embryonic development in a culture system supplemented with L-cysteine.

Various methods have been used to remove reactive oxygen species (ROS) generated from in vitro culture (IVC) conditions that can cause cell injury or death, including the application of low oxygen (O(2)) tension and the addition of antioxidants. The beneficial effects of antioxidants and O(2) tension on IVC of porcine embryos, however, are controversial among researchers. In this study, we sought to determine the effects and optimal concentrations of antioxidants for the development of porcine embryos in an IVC system. Specifically, we examined the synergistic effects of antioxidants on development to the blastocyst stage in a culture system supplemented with L-cysteine during IVM. Of the antioxidants tested (melatonin, glutathione (GSH), ß-mercaptoethanol (ß-ME), N-acetylcysteine (NAC) and dithiothreitol (DTT)), addition of GSH (1 mM) or ß-ME (25 µM) significantly increased development to the blastocyst stage compared with the controls without antioxidant treatment (22.2 ± 4.2% for 1 mM GSH, 25.9 ± 2.2% for 25 µM ß-ME and 12-13% for the control, P<0.05). In addition, the mean cell number per blastocyst was increased by approximately 1.7-fold in the presence of GSH or ß -ME. These GSH- and ß-ME-induced increases in development to the blastocyst stage and total cell number, however, were not mimicked by melatonin, NAC or DTT, all of which are ROS scavengers. The combination of GSH or ß-ME with L-cysteine significantly reduced high O(2) tension-induced ROS production (P<0.05). These results suggest that a combination of 1 mM GSH or 25 µM ß-ME with 1 mM L-cysteine could be used for production of high quality porcine blastocysts in IVC systems.

Effect of estradiol benzoate or GnRH treatment prior to superstimulation in CIDR-treated, Korean native cows (Bos taurus).

The objective of this study was to evaluate the effectiveness of superovulatory protocols by synchronizing the emergence of the follicular wave using estradiol benzoate (EB) or GnRH in CIDR-treated, Korean cows. Sixty-six cows were used in the study and these were divided into three groups. The standard group comprised cows that were between days 8 and 12 of their estrous cycle (n=22). The remaining 44 cows, at all other stages of the estrous cycle, received CIDR and were assigned to two treatment groups that received either 2mg EB (EB-CIDR group, n=22) or 100 microg GnRH (GnRH-CIDR group, n=22) 1 day after CIDR insertion. Gonadotropin treatment began between the 8th and 12th days of the estrous cycle in the standard group, 5 days after EB injection in the EB-CIDR group, and 3 days after GnRH injection in the GnRH-CIDR group. All cows were superovulated with porcine FSH (pFSH) twice daily, with the dose (total 28 mg) decreasing gradually over 4 days. On the 5th and 6th injections of pFSH, 25 and 15 mg doses of PGF(2alpha) were administered. CIDR was withdrawn at the 7th pFSH injection and the cows received 200 microg GnRH at 24h after CIDR withdrawal. Cows were artificially inseminated twice at 36 and 48 h post-CIDR withdrawal and embryos were recovered 7 days after the 1st insemination. The numbers of preovulatory follicles (22.9-28.2), ovulated preovulatory follicles (17.6-21.7) and CL (15.9-17.9) detected by ultrasonography did not differ among groups (P>0.05). Similarly, the numbers of total ova (6.7-10.0), transferable embryos (4.0-6.0), degenerate embryos (1.1-1.8) and unfertilized ova (1.3-4.3) did not differ among groups (P>0.05). Progesterone and estradiol concentrations during superovulation treatments and at embryo recovery were also the same in all groups (P>0.05). We conclude that in CIDR-treated Korean native cows, superovulatory treatments that follow administration of either EB or GnRH (at any stage of the estrous cycle) result in both a superovulatory response and embryo yield comparable to conventional superovulation protocols.

Expression and regulation of interleukin 6 and its receptor at the maternal-conceptus interface during pregnancy in pigs.

It has been well established that interleukin 6 (IL6), a pleiotropic cytokine with multiple functional roles, is widely expressed in the female reproductive tract and mediates blastocyst implantation and placental development in many species. Uterine expression of IL6 during early pregnancy has been studied in pigs, but expression and function of IL6 at the maternal-placental interface throughout pregnancy have not been determined. Thus, we examined expression of IL6 and its receptors, IL6 receptor (IL6R) and GP130, in the uterine endometrium on Days 12 and 15 of the estrous cycle, and Days 12, 15, 30, 60, 90, and 114 of pregnancy, conceptus on Days 12 and 15, and chorioallantoic tissues on Days 30, 60, 90, and 114 of pregnancy in pigs. The expression of IL6, IL6R, and GP130 mRNA in the endometrial tissues increased dramatically during mid-to late-pregnancy and decreased at term. IL6, IL6R, and GP130 mRNAs were also expressed in conceptus and chorioallantoic tissues. Expression of IL6 mRNA was mainly localized to endometrial epithelial and stromal cells and chorioallantoic tissues, while IL6R and GP130 mRNAs were localized to glandular epithelial cells during pregnancy. The expression of IL6 mRNA was decreased by estrogen and progesterone treatment, whereas increasing doses of IL1ß induced the expression of IL6 mRNA, but not IL6R and GP130 mRNAs, in endometrial tissue explants. These results indicate that expression of IL6 and its receptors at the feto-maternal interface is regulated in a stage- and cell-type-specific manner during pregnancy, suggesting that IL6 and its receptor signaling system may play an important role in the maintenance of pregnancy in pigs.

Novel Acinetobacter parvus HANDI 309 microbial biomass for the production of N-acetyl-ß-d-glucosamine (GlcNAc) using swollen chitin substrate in submerged fermentation.

BACKGROUND: N-acetyl-ß-d-glucosamine (GlcNAc)6 is extensively used as an important bio-agent and a functional food additive. The traditional chemical process for GlcNAc production has some problems such as high production cost, low yield, and acidic pollution. Therefore, to discover a novel chitinase that is suitable for bioconversion of chitin to GlcNAc would be of great value. RESULTS: Here, we describe the complete isolation and functional characterization of a novel exo-chitinase from Acinetobacter parvus HANDI 309 for the conversion of chitin. The identified exo-chitinase mainly produced N-acetyl-d-glucosamine, using chitin as a substrate by submerged fermentation. The A. parvus HANDI 309 biofuels producing exo-chitinase were characterized by TLC, and was further validated and quantified by HPLC. Furthermore, the optimal temperature and pH for the exo-chitinase activity was obtained in the culture conditions of 30 °C and 7.0, respectively. The maximum growth of the stationary phase was reached in 24 h after incubation. These results suggest that A. parvus HANDI 309 biofuels producing exo-chitinases may have great potential in chitin to N-acetyl-d-glucosamine conversion. CONCLUSIONS: The excellent thermostability and hydrolytic properties may give the exo-chitinase great potential in chitin to GlcNAc conversion in industry. This is the first report that A. parvus HANDI 309 is a novel bacterial strain that has the ability to produce an enormous amount of exo-chitinase-producing bio-agents in a short time on an industrial scale without any pretreatment, as well as being potentially valuable in the food and pharmaceutical industries.

Exploring the binding nature of pyrrolidine pocket-dependent interactions in the polo-box domain of polo-like kinase 1.

BACKGROUND: Over the years, a great deal of effort has been focused on the design and synthesis of potent, linear peptide inhibitors targeting the polo-like kinase 1 (Plk1), which is critically involved in multiple mitotic processes and has been established as an adverse prognostic marker for tumor patients. Plk1 localizes to its intracellular anchoring sites via its polo-box domain, and inhibiting the Plk1 polo-box domain has been considered as an approach to circumvent the specificity problems associated with inhibiting the conserved adenosine triphosphate-binding pocket. The polo-box domain consists of two different binding regions, such as the unique, broader pyrrolidine-binding pocket and the conserved, narrow, Tyr-rich hydrophobic channel, among the three Plk polo-box domains (Plks 1-3), respectively. Therefore, the studies that provide insights into the binding nature of the unique, broader pyrrolidine-binding pocket might lead to the development of selective Plk1-inhibitory compounds. METHODOLOGY/PRINCIPAL FINDINGS: In an attempt to retain the monospecificity by targeting the unique, broader pyrrolidine-binding pocket, here, for the first time, a systematic approach was undertaken to examine the structure-activity relationship of N-terminal-truncated PLHSpTM derivatives, to apply a site-directed ligand approach using bulky aromatic and non-aromatic systems, and to characterize the binding nature of these analogues using X-ray crystallographic studies. We have identified a new mode of binding interactions, having improved binding affinity and retaining the Plk1 polo-box domain specificity, at the pyrrolidine-binding pocket. Furthermore, our data revealed that the pyrrolidine-binding pocket was very specific to recognize a short and bulky hydrophobic ligand like adamantane, whereas the Tyr-rich hydrophobic channel was specific with lengthy and small hydrophobic groups. CONCLUSION/SIGNIFICANCE: The progress made using our site-directed ligands validated this approach to specifically direct the ligand into the unique pyrrolidine-binding region, and it extends the applicability of the strategy for discovering selective protein-protein interaction inhibitors.

Discovery of torezolid as a novel 5-hydroxymethyl-oxazolidinone antibacterial agent.

A series of novel substituted pyridyl phenyl oxazolidinone analogues were synthesized and their structure-activity relationship (SAR) was investigated based on in vitro and in vivo antibacterial activities. The minimum inhibitory concentrations (MICs) of the synthesized compounds against methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE) ranged from 0.12 to 2.0 µg/mL, and against Haemophilus influenzae (Hi) from 2.0 to 8.0 µg/mL. Compared to linezolid, only four compounds (11, 12, 21 and 29) showed higher in vitro antibacterial activities and better in vivo protective effects in mice. To improve the aqueous solubility, various prodrugs of compound 11 (DA-7157), which exerted a potency that was enhanced by 2-8-fold compared to that of linezolid, were synthesized. Among the prodrugs, the phosphate compound 42 exhibited excellent aqueous solubility (>50mg/mL in DW) and good pharmacokinetic profiles, along with better in vivo efficacy than linezolid. This compound 42 is currently undergoing clinical trials with the brand name Torezolid.

The effect of reduced dose and number of treatments of FSH on superovulatory response in CIDR-treated Korean native cows.

The objective of this study was to investigate the effect of dosage and number of days of follicle stimulating hormone (FSH) treatment on superovulatory response in controlled internal drug release (CIDR)-treated Korean native cows. Forty cows underwent two superovulatory treatments with a crossover design. Cows, at random stages of the estrous cycle, received a CIDR together with injections of 1 mg estradiol benzoate and 50 mg progesterone, and gonadotropin treatment began 4 days later. The cows were divided into 2 groups based on the dosage and number of days of treatment with porcine FSH; a total of 28 mg FSH was given in twice daily intramascular injections in decreasing doses over 4 days (5, 5, 4, 4, 3, 3, 2 and 2 mg; T1 group, n=20) or a total of 24 mg FSH was given in twice daily decreasing doses over 3 days (5, 5, 4, 4, 3 and 3 mg; T2 group, n=20). This was followed by the alternate treatment in the subsequent superovulation. The cows were treated identically in all other respects. PGF(2alpha) (25 mg and 15 mg) was given with the 5th and 6th injections of FSH, CIDR were withdrawn at the 6th FSH injection and the cows received 200 microg GnRH 36 h after CIDR withdrawal. The cows were artificially inseminated twice, at 48 and 60 h after CIDR withdrawal, using commercial semen from four Korean native bulls, and embryos were recovered 6 or 7 days after the 2nd insemination. The numbers of corpora lutea (CL; 7.9+/-1.0 vs. 8.3+/-1.1) and large follicles (1.2+/-0.2 vs. 1.3+/-0.3) present at the time embryo recovery, as detected by ultrasonography, did not differ between the T1 and T2 groups (P>0.05). Similarly, the numbers of total ova/embryos (6.2+/-0.9 vs. 6.4+/-1.1), transferable embryos (3.4+/-0.8 vs. 3.2+/-0.7), degenerate embryos (0.8+/-0.2 vs. 1.0+/-0.3) and unfertilized ova (2.1+/-0.5 vs. 2.2+/-0.5) did not differ between the groups (P>0.05). These data indicate that a reduced dose (24 vs. 28 mg) and number of treatments (6 vs. 8) of FSH for superovulation of CIDR-treated Korean native cows does not affect the embryo yield.

A CIDR-based timed embryo transfer protocol increases the pregnancy rate of lactating repeat breeder dairy cows.

This study evaluated the pregnancy rates following either a controlled internal drug release (CIDR)-based timed artificial insemination (TAI) or an embryo transfer (TET) protocol compared with that following a single PGF(2alpha) injection and AI after estrus (AIE) in lactating repeat breeder dairy cows. Fifty-three lactating dairy cows diagnosed as repeat breeders were used in this study and were randomly assigned to the following three treatments. (1) Cows, at random stages of the estrous cycle, received a CIDR device and 2 mg estradiol benzoate (EB; Day 0), a 25 mg PGF(2) (alpha) injection at the time of CIDR removal on Day 7 and a 1 mg EB injection on Day 8. The cows then received TAI 30 h (Day 9) after the second EB injection using dairy semen (TAI group, n=13). (2) Cows, at random stages of the estrous cycle, received the same hormonal treatments as in the TAI group. The cows then received TET on Day 16 using frozen-thawed blastocysts or morula embryos collected from Korean native cattle donors (TET group, n=13). (3) Cows, at the luteal phase, received a 25 mg injection of PGF(2alpha) and AIE using dairy semen (control group, n=27). The ovaries of the cows in the TET group were examined by transrectal ultrasonography to determine ovulation of the preovulatory follicles, and blood samples were collected for serum progesterone (P4) analysis. The pregnancy rate was significantly higher in the TET group (53.8%) than in the control (18.5%) or TAI (7.7%) groups (P<0.05). The ultrasonographic observations demonstrated that all the cows in the TET group ovulated the preovulatory follicles and concomitantly formed new corpora lutea. Accordingly, the mean serum P4 concentration remained constant between Day 0 and Day 7 of the luteal phase, decreased dramatically on Day 8 (P<0.01) and subsequently increased by Day 16 (P<0.01). These data suggest that the CIDR-based TET protocol can be used to effectively increase the pregnancy rate in lactating repeat breeder dairy cows.

Effect of exposure duration of ovaries and oocytes at ambient temperature on parthenogenetic development of porcine follicular oocytes.

This study was conducted to evaluate the effects of exposing porcine ovaries to 30-33 C during transportation for 4 h and subsequently room temperature (25 C) for 6 h of storage on in vitro maturation (IVM) and subsequent parthenogenetic development of oocytes collected from the ovaries. After IVM, oocytes having a tight oopalsm membrane and no signs of degeneration were exposed to Dulbecco's phosphate-buffered saline (DPBS) with 7% ethanol (v/v) for 7 min to induce parthenogenetic activation. Moreover, we also determined whether exposure of the collected oocytes to room temperature for 1, 2 and 4 h in DPBS or porcine follicular fluid (pFF) affected parthenogenetic development. When porcine ovaries were stored after transportation, oocytes collected from the stored ovaries showed a significantly higher rate of degeneration after 65 h of IVM (58.4%) and a significantly lower rate of cleavage after parthenogenetic activation (40.1%) than oocytes collected from ovaries immediately after transportation (38.9% and 47.4%, respectively). However, there was no significant difference in developmental rates to the morula and blastocyst stages between these two groups (14.4% and 14.3%, respectively). The duration of preservation, 1, 2, and 4 h, of oocytes in DPBS did not affect parthenogenetic development. In contrast, when preserved for 4 h in pFF, the developmental rates of the oocytes were significantly decreased. This suggested that some factor(s) in follicular fluid affects the developmental rate of oocytes with the passage of time in ambient conditions. These results suggest that even after 6 h storage of ovaries, oocytes having normal morphology after IVM have the same rate of parthenogenetic development as oocytes collected from ovaries just after 4 h of transportation, except for a lower cleavage rate, and that exposure of oocytes to room temperature for 4 h in DPBS does not affect their parthenogenetic developmental competence.

Pronucleus formation, DNA synthesis and metaphase entry in porcine oocytes following intracytoplasmic injection of murine spermatozoa.

The onset of pronucleus formation and DNA synthesis in porcine oocytes following the injection of porcine or murine sperm was determined in order to obtain insights into species-specific paternal factors that contribute to fertilisation. Similar frequencies of oocytes with female pronuclei were observed after injection with porcine sperm or with murine sperm. In contrast, male pronuclei formed 8-9 h following the injection of porcine sperm, and 6-8 h following the injection of murine sperm. After pronucleus formation maternally derived microtubules were assembled and appeared to move both male and female pronuclei to the oocyte centre. A few porcine oocytes entered metaphase 22 h after the injection of murine sperm, but normal cell division was not observed. The mean time of onset of S-phase in male pronuclei was 9.7 h following porcine sperm injection and 7.4 h following mouse sperm injection. Ultrastructural observation revealed that male pronuclei derived from murine sperm in porcine oocytes are morphologically similar to normal male pronuclei in porcine zygotes. These results suggest that species-specific paternal factors influence the onset of pronucleus formation and DNA synthesis. However, normal nuclear cytoplasmic interactions were observed in porcine S-phase oocytes following murine sperm injection.