Molecular cloning of porcine growth differentiation factor 9 (GDF-9) cDNA and its role in early folliculogenesis: direct ovarian injection of GDF-9 gene fragments promotes early folliculogenesis

Growth differentiation factor-9 (GDF-9) is a growth factor secreted by oocytes in growing ovarian follicles. To investigate the ovarian function of GDF-9 in pigs, we first cloned porcine GDF-9 complementary DNA (cDNA), and then injected its gene fragments into the ovary in gilts. Porcine GDF-9 has open reading frame (ORF) homologies of 81.4%, 84.6%, 84.2%, 72.7% and 72.6% with its human, bovine, ovine, rat and mouse counterparts respectively. Regarding the deduced amino-acid sequence of the mature protein, the corresponding homologies reach 92.1%, 97.8%, 97.0%, 89.6% and 88.1% respectively. To investigate the role of GDF-9 in early folliculogenesis, the ovaries of 2-month-old prepubertal gilts were injected with GDF-9 gene fragments. The injection of porcine GDF-9 gene fragments resulted in an increase in the number of primary, secondary and tertiary follicles, concomitant with a decrease in the number of primordial follicles. These results indicated that exogenous GDF-9 can promote early folliculogenesis in the porcine ovary, and that a technique for direct ovarian injection of GFD-9 gene fragments may contribute to a novel therapy for prevention and treatment of infertility associated with ovarian dysfunction.     

The protein encoded by cancer/testis gene D40/AF15q14 is localized in spermatocytes, acrosomes of spermatids and ejaculated spermatozoa

We have previously identified and cloned a human gene, D40, that is preferentially expressed in testis among normal organs, while it is widely expressed in various human tumor cell lines and primary tumors derived from different organs. In this report, we have examined the expression and localization of this protein in human testis with an antibody specific to D40 protein. In Western analyses, the anti-D40 antibody recognized a major band with a molecular mass of 300 kDa and a minor band of 250 kDa. These bands were not observed in the testis lysates from patients with Sertoli-cell-only syndrome and with Kleinfelter syndrome, who lack germ cells of the testis, indicating that D40 protein is expressed in the germ cells of normal testis. Immunohistochemical studies have revealed that D40 protein is highly expressed in spermatocytes and in the pre-acrosome of round spermatids. In the acrosome, D40 protein expression is observed not inside but outside the acrosome membrane. This is consistent with the finding that the amino-acid sequence at the amino terminal of the D40 protein lacks a hydrophobic signal peptide that is required for proteins to translocate to the membrane. Expression of D40 protein is observed in the acrosome of ejaculated spermatozoa as well, although the level is low compared with that in the pre-acrosome of spermatids. These results suggest that D40 protein plays important roles in spermatogenesis, especially in the formation and maintenance of the acrosome.     

Lipid analog with 2-nitrophenol trigger designed for liposome fusion at physiological pH

A novel lipid analog with two long alkyl (C₁₆) chains, an aspartate skeleton, a connecting alkyl (C₈) chain, and 2-nitrophenol trigger group is synthesized by an efficient synthetic route, which can induce liposome fusion at physiological pH.     

Preparation of carbon microparticle assemblies from phenolic resin using an inverse opal templating method

Three-dimensionally (3D) well-ordered carbon microparticle assemblies with different particle morphologies were fabricated by infiltration of phenolic resin solution into SiO₂ inverse opal structures and subsequent carbonization. The effect of the phenolic resin solution concentration and the carbonization temperature on the morphology of the fabricated carbon microparticles was investigated. At a carbonization temperature of 1000 °C, carbon microparticles with interlocked bridges were obtained when the concentration of phenolic resin solution is 40 wt% and hollow carbon microparticles with opened window channels were obtained at a concentration of 30–35 wt%. When the carbonization temperature was decreased to 500 °C, carbon microparticles with interlocked bridges also were observed, even when the phenolic resin concentration was 30 wt%. The structures and properties of the carbon microparticles and their assemblies were characterized using SEM, XRD, and N₂ adsorption.     

Synthesis and properties of ellipsoidal hematite/silicone core-shell particles

Monodispersed ellipsoidal hematite particles were synthesized and coated with silicone using a silane-coupling agent, phenyltriethoxysilane. The thickness of the silicone shell was controlled by the concentration of phenyltriethoxysilane and the presence of n-hexadecyltrimethylammonium chloride, which was thought to modify the surface properties of the hematite particles to be organophilic. The hematite/silicone core-shell particles were strongly hydrophobic and had a good dispersibility and stability in toluene. Hollow ellipsoidal silicone particles were obtained by the dissolution of hematite with hydrochloric acid from the core-shell particles.     

Determination of the thermal conductivity of argon and nitrogen over a wide temperature range through data evaluation and shock-tube experiments

Reliable and well-established methods to measure the thermal conductivity of gases are available only in the moderate temperature range, namely, up to about 1000 K. In the present study, a set of the most probable thermal conductivity values of components of gaseous combustion products in a wide range of temperatures has been obtained through an optimum combination of three procedures: critical assessment of available data in the moderate temperature range, experimental determination by the shock-tube method at high temperatures, and theoretacal estimation of temperature dependence in the intermediate temperature range. Among the components of combustion products, one monatomic gas and one diatomic gas, namely, argon and nitrogen, were studied in the present paper. The shock-tube measurements have been performed in the temperature ranges 1000–4500 K for argon and 500–2200 K for nitrogen. The results of the critical evaluation and the shock-tube measurements have been combined with the aid of theoretically assumed temperature dependence of the thermal conductivity.Invited paper presented at the Ninth Symposium on Thermophysical Properties, June 24–27, 1985, Boulder, Colorado, U.S.A.     

A new detection method for the primary cosmic heavy nuclei by the plastic track detector CR-39

A new multiple scanning method is developed and applied to detect cosmic heavy nuclei. The plastic track detectors CR-39 exposed to cosmic heavy nuclei are over-etched in 8.0N NaOH for 94 h at 80°C. We found that the etched cones can easily be scanned on the plastic plates without using an optical microscope. The trajectories of heavy nuclei can be reconstructed with the naked eyes by superposing four or more over-etched plates. The charge detection threshold is obtained by this method, and appears to be 10. The charge resolution is estimated to be 1.0 charge units for iron nuclei. By adopting this method, the scanning time is much shortened and CR-39 plastic chamber size can easily be scaled up. The method is very useful for detecting particles with low flux intensity, like for searching magnetic monopoles or for measuring the high energy spectra of cosmic heavy nuclei.     

Determination of bisphenol A in canned vegetables and fruit by high performance liquid chromatography

A high performance liquid chromatograph y (HPLC) method was developed for the determination of bisphenol A (BPA) that had migrated into canned fruit and vegetables. BPA was extracted with acetonitrile from the solid portion of canned food, and with an OASIS HLB cartridge from the aqueous portion, respectively. Both extracts were cleaned up on a Florisil cartridge. The HPLC separation was carried out on a Wakosil II 3C18 RS column (4.6 × 150mm) with acetonitrile-water (40:60, v/v) as a mobile phase with a flow rate of 0.8 ml/min. BPA was detectable by UV detector at 228 nm and determined with the similarity of chromatographic peak spectrum by multiwavelength detector (similarity index was 0.99 or above). The quantification limits were 10 ng/g for the solid portion and 5 ng/ml for the aqueous portion, respectively. BPA was mainly detected in the solid portion of canned food and found at the maximum level of 11 μg per can. To verify migration into the solid portion of canned food, a partitioning experiment was carried out.     

A reinforcement learning with switching controllers for a continuous action space

Reinforcement learning (RL) attracts much attention as a technique for realizing computational intelligence such as adaptive and autonomous decentralized systems. In general, however, it is not easy to put RL to practical use. This difficulty includes the problem of designing a suitable action space for an agent, i.e., satisfying two requirements in trade-off: (i) to keep the characteristics (or structure) of an original search space as much as possible in order to seek strategies that lie close to the optimal, and (ii) to reduce the search space as much as possible in order to expedite the learning process. In order to design a suitable action space adaptively, in this article, we propose a RL model with switching controllers based on Q-learning and an actor-critic to mimic the process of an infant’s motor development in which gross motor skills develop before fine motor skills. Then a method for switching controllers is constructed by introducing and referring to the “entropy.” Further, through computational experiments by using a path-planning problem with continuous action space, the validity and potential of the proposed method have been confirmed.