Porcine spermatogonial stem cells (SSCs) prefer three-dimensional (3D) culture systems to 2D ones for the maintenance of self-renewal. Of the many 3D culture systems, agar-based hydrogels are candidates for supporting porcine SSC self-renewal, and there are various types of agar powder that can be used. In this study, we sought to identify an agar-based 3D hydrogel system that exhibited strong efficacy in the maintenance of porcine SSC self-renewal. First, 3D hydrogels with different mechanics were prepared with various concentrations of Bacto agar, lysogeny broth (LB) agar, and agarose powder, and the 3D hydrogel with the strongest alkaline phosphatase (AP) activity and greatest increase in colony size was identified for the different types of agar powder. Second, among the porcine SSCs cultured in the different 3D hydrogels, we analyzed the colony formation, morphology, and size; AP activity; and transcription and translation of porcine SSC-related genes, and these were compared to determine the optimal 3D hydrogel system for the maintenance of porcine SSC self-renewal. We found that 0.6% (w/v) Bacto agar-, 1% (w/v) LB agar-, and 0.2% (w/v) agarose-based 3D hydrogels showed the strongest maintenance of AP activity and the most pronounced increase in colony size in the culture of porcine SSCs. Moreover, among these hydrogels, the strongest transcription and translation of porcine SSC-related genes and largest colony size were detected in porcine SSCs cultured in the 0.2% (w/v) agarose-based 3D hydrogel, whereas there were no significant differences in colony formation and morphology. These results demonstrate that the 0.2% (w/v) agarose-based 3D hydrogel can be effectively used for the maintenance of porcine SSC self-renewal.

In all mammalian species, progesterone is essential in the preparation for and maintenance of pregnancy, if it occurs. Progesterone primes the endometrium for possible implantation and inhibits uterine contraction until birth. Aldo-keto reductases (AKRs) belong to a superfamily of NADPH-dependent reductases that act on a wide range of substrates, including simple carbohydrates, steroid hormones, and endogenous prostaglandins. 20-alpha hydroxysteroid dehydrogenase (20α-HSD; EC.1.1.1.149) enzyme belongs to the family of aldo-keto reductases. 20α-HSD predominantly converts progesterone into its biologically inactive form 20α-hydroxyprogesterone (20α -OHP), and plays a crucial role in the termination of pregnancy and initiation of parturition. In addition, the activity of 20α-HSD during the luteal phase is known to be inhibited by prolactin. We have been reporting on the molecular characterizations of placental and ovarian 20α-HSD in the bovine, pig, deer and monkey. In this study, we focused on the 20α-HSD expression in testis(6, 9, 12, 18 and 21 days after birth) of miniature pig. The protein expression and localization were detected by Western blotting and Immunohistochemical analysis. 20α-HSD protein was detected at molecular weight of 37-kDa by Western blot analysis. Also the RNA expression were detected by Reverse Transcription-PCR and quantification PCR. Additionally, We are going to analysis the function and role of 20α-HSD in the pig testis.

Zika virus, mosquito-borne disease, caused by mosquitoes has been increased the importance. From March to September, twice a month from 7 different points (3 residential areas, 3 migratory bird sanctuary and 1 cattle shed) were collected using BG trap and BL trap. After identifying the mosquitoes collected, we confirmed the virus infection. Total 26,531 mosquitoes (6 genus 9 species) were collected, virus has been detected from the 3 species (Aedes vexans, Cuilex tritaeniorhynchus, and Amigeres subalbatus) of mosquitoes of them. It showed the highest peak in August, and then gradually decreased. The most common mosquito species was collected Aedes vexan (16,637) in the cattle shed.

Despite many researches related with in-vitro culture of porcine spematogonial stem cells (SSCs), adherent culture system widely used has shown a limitation in the maintenance of porcine SSC self-renewal. Therefore, in order to overcome this obstacle, suspension culture, which is known to have numerous advantage over adherent culture, was applied to the culture of porcine SSCs. Porcine SSCs retrieved from neonatal testes were suspension-cultured for 5 days or 20 days, and characteristics of suspension-cultured porcine SSCs including proliferation, alkaline phosphatase (AP) activity, and self-renewal-specific gene expression were investigated and compared with those of adherent-cul-tured porcine SSCs. As the results, the suspension-cultured porcine SSCs showed entirely non-proliferative and significantly higher rate of AP-positive cells and expression of self-renewal-specific genes than the adherent-cultured porcine SSCs. In addition, long-term culture of porcine SSCs in suspension condition induced significant decrease in the yield of AP staining-positive cells on post-day 10 of culture. These results showed that suspension culture was inappropriate to culture porcine SSCs, because the culture of porcine SSCs in suspension condition didn’t stimulate proliferation and maintain AP activity of porcine SSCs, regardless of culture periods.

Background : 1-Deoxynojirimycin (DNJ) is a representative compound of the antidiabetic constituent in mulberry leaves (Morus alba L.). The hot water extracts of mulberry leaves were fermented with lactic acid bacteria in order to analysis the changes of the DNJ contents and α-glucosidase inhibition. Methods and Results : The mulberry leaves were extracted with hot water (121℃, 3 hr). The extracts were fermented with nine strains of lactic acid bacteria such as Lactobacillus acidophilus, in order to increase the contents of DNJ and α-glucosidase inhibition. DNJ contents in the fermented extracts were determinated by HPLC analysis. The α-glucosidase inhibition was mesured by comparing dose-inhibition curves of α-glucosidase and IC50 value. The DNJ contents after fermentation have increased in the almost fermented extracts. Especially, DNJ of the extracts fermented with L. acidophilus was increased from 8.38 ㎍ ㎖ -1 to 21.77 ㎍ ㎖-1. IC50 values of the α-glucosidase inhibition were shown to be decreased in the fermented extracts. The extracts fermented with L. casei KCTC 3109 was determined at 290.04 ㎍ ㎖-1, resulting it is lower about 140 ㎍ ㎖-1 than 429.76 ㎍ ㎖-1 of the control. Conclusion : From the above results, we may suggest that the lactic acid fermentation of mulberry leaves extracts can more enhance the hypoglycemic activities such as DNJ contents.

Background : The hypoglycemic effects of mulberry leaves extract were evaluated by comparing the abilities on glucose uptake in C2C12 myotubes and 3T3-L1 adipocyte. Glucose uptake of the extracts were identified to be enhanced by bio-conversion using cellulolytic enzyme like Viscozyme. Methods and Results : The mulberry (Morus alba L) leaves were extracted with 30% ethanol or hot water. The hypoglycemic compounds such as Moracin C and, Quercetin and 1-Deoxynojirimycin were identified from the extracts of mulberry leaves. The extracts were fermented using kinds of celluolytic enzymes, which were vicozyme, pectinase, β-glucosidase and xylanase, in order to increase the contents of hypoglycemic constituents in the extracts. The hypoglycemic effects of the fermented extracts were evaluated by comparing the abilities on glucose uptake in C2C12 myotubes and 3T3-L1 adipocyte. The extracts of mulberry leaves fermented with only Viscozyme were identified to increase glucose uptake in C2C12 myotubes and 3T3-L1 adipocyte by supplement of the concentration of 10 μM extracts, compared to insulin as control. However, bio-conversion effects by other enzymes were not shown in the treatments, suggesting hypoglycemic constituents in the extracts of mulberry leaves can be conversed to more active compounds by cellulolytic enzyme treatment like viscozyme. Conclusion : From the above results, we may suggest that the hypoglycemic constituents in mulberry leaves extracts can be conversed to more active compounds by cellulolytic enzymes.

The vascular system of plants consists of two conducting tissues, xylem and phloem, which differentiate from procambium cells. Xylem serves as a transporting system for water and signaling molecules and is formed by sequential developmental processes, including cell division/expansion, secondary cell wall deposition, vacuole collapse, and programmed cell death (PCD). PCD during xylem differentiation is accomplished by degradation of cytoplasmic constituents, and it is required for the formation of hollow vessels, known as tracheary elements (TEs). Our recent study revealed that the small GTPase RabG3b acts as a regulator of TE differentiation through its autophagic activation. By using an Arabidopsis in vitro cell culture system, we showed that autophagy is activated during TE differentiation. Overexpression of a constitutively active RabG3b (RabG3bCA) significantly enhances both autophagy and TE differentiation, which are consistently suppressed in transgenic plants overexpressing a dominant negative form (RabG3bDN) or RabG3bRNAi (RabG3bRNAi), a brassinosteroidinsensitive mutant bri1-301, and an autophagy mutant atg5-1. Wood (called secondary xylem) is the most abundant biomass produced by land plants including Populus and Eucalyptus, and therefore is considered to be one of the most cost-effective and renewable bioenergy resources. In an attempt to enhance xylem differentiation and thus to improve biomass traits in poplars, we generated transgenic poplars overexpressing the RabG3bCA form. As notable phenotypes, both stem height and diameter were increased and xylem area in vascular bundles was significantly expanded in RabG3bCA transgenic poplars compared to control plants. Taken together, these results demonstrate that RabG3b regulates xylem differentiation in both Arabidopsis and Populus. This study enhances our understanding of biological mechanisms underlying wood formation and serve as a framework to engineer the quality and quantity of wood as useful biomass.

Ionizing radiation is known to cause chromosomal alterations such as inversions and deletions and affects gene expression within the plant genome. To monitor the genome-wide transcriptome changes by ionizing radiation, we used rice Affimetrix GeneChip microarray to identify genes that are up- or down regulated by gamma-ray (200 Gy, 60Co source), cosmic-ray and ion beam (40 Gy, 220 MeV carbon ion). The overall expression patterns between gamma-ray and ion beam were similar but cosmic-ray was regulated differently. Combined results from all 3 radiations identified 27 up-regulated genes and 188 down regulated genes. These results mean the induction of similar mechanism changes in treatments of gamma ray and ion beam. However the different expression in treatment of cosmic-ray might be due to the other environmental conditions. Among the commonly up- or down- regulated genes, we chose highly up- or down- regulated several genes and confirmed its regulation in response to ionizing radiation exposure by RT-PCR analysis. Moreover, we showed that specific co-expression networks of candidate radio marker genes by ARACNE algorithm. Our results present profiles of gene expression related to different ionizing radiation and marker gene to predict sensitivity to ionizing radiation, such as GS (glutelin subunit) and FBX322.

This study is to survey the ultrastructure of gamete cells and micropyle of pre-fertilized and post-fertilized eggs after HCG hormone treatment by transmission and scanning electron microscopy (TEM and SEM) in E. akaara, E. bruneus and E. septemfasciatus. These fishes are economical importance species for Jeju coastal area resources. In spite of its an importance resources, details studies on the ultrastructural aspects of gamete cells for its reproductive biology have not been undertaken. Morphological features of ovulation process have been studied during its normal occurrence in the reproductive cycle of these fish by light microscopy. Moreover, it has been studied for many years to induce spawning by environmental factors (day length, water temperature etc) or injection of HCG for ovulation in these species. Studies on the micropyle was mainly focused on the eggs of insects, fresh water and a few sea water fishes. Micropylar structure of fish displays morphological characteristics of interspecies-specific by inhabitant environment and spawning feature. On the other hand, it is an importance cue for a taxonomical indicator and identification fish eggs. SEM studies were performed on growing and mature oocytes obtained by stripping and cannulation from 3 grouper species sampled between July and August in spawning season. The outer layer of chorion of preovulatory growing stage oocytes could be divided into four layers; zona pellucida, follicular cell layers consisted of granulosa and thecal cells layer and the most outer ovigerous lamella. Ovulation process of mature stage oocytes initiated by rupture of ovigerous lamella and ovulated by contraction of follicular cell layers. Besides, the micropylar shape of ripe stage oocytes in E. akaara, E. bruneus and E. septemfasciatus presented volcano or crateriform-like cylindrical form. Internal structure of micropylar vestibule displayed cylindrical clockwise 8 or 10 spiral arrangement structure in these species. The micropyle diameter and apparatus at the animal pole differ significantly among the 3 species. The difference in their diameters suggests species-specific in the correlation between spermatozoal head size and micropylar diameter for polyspermy prevention and hybridization during fertilization. Besides, after artificial fertilization, the vestibule morphologically transformed into dom-shape and pillar-shape for fertilization cone formation. Pores of various sizes in the 3 grouper species were somewhat regularly distributed in concentric circles only around the micropyle. In particular, large pores had numerous gill filament-shaped projections connected to oolemma. These structures are suggested to be related to gas exchange, osmoregulation, and micronutrient influx or efflux between eggs and water during fertilization and egg development. In addition, spermatozoa ultrastructure was examined in 3 grouper species. TEM investigation revealed that, in all species, spermatozoa display a round head, a nucleus containing highly condensed, filamentous chromatin clusters, no acrosome, a short midpieces consisting of numerous mitochondria and the proximal and distal centrioles and a flagellum exhibiting the typical axoneme structure (9+2). Especial, both E. akaara and E. bruneus display regular laternal fins in flagella, but in E. septemfasciatus, no fins in flagella with hook shape tails.