With about 80% of the global economy expected to shift to the global market by 2030, exports of reverse direct purchase products, in which foreign consumers purchase products from online shopping malls in Korea, are growing 55% annually. As of 2021, sales of reverse direct purchases in South Korea increased 50.6% from the previous year, surpassing 40 million. In order for domestic SMEs(Small and medium sized enterprises) to enter overseas markets, it is important to come up with export strategies based on various market analysis information, but for domestic small and medium-sized sellers, entry barriers are high, such as lack of information on overseas markets and difficulty in selecting local preferred products and determining competitive sales prices. This study develops an AI-based product recommendation and sales price estimation model to collect and analyze global shopping malls and product trends to provide marketing information that presents promising and appropriate product sales prices to small and medium-sized sellers who have difficulty collecting global market information. The product recommendation model is based on the LTR (Learning To Rank) methodology. As a result of comparing performance with nDCG, the Pair-wise-based XGBoost-LambdaMART Model was measured to be excellent. The sales price estimation model uses a regression algorithm. According to the R-Squared value, the Light Gradient Boosting Machine performs best in this model.

The present study was aimed to estimate the effect of ensiling period and bacterial inoculants on chemical compositions and fermentation characteristics on rye silage harvested at delayed stage. Rye (Secale cereale L.) was harvested after 20 days of heading stage (29.4% dry matter, DM). The harvested rye forage was applied with different inoculants following: applications of distilled water (CON), Lactobacillus brevis (LBB), Leuconostoc holzapfelii (LCH), or mixture of LBB and LCH at 1:1 ratio (MIX). Each forage was ensiled into 20 L mini bucket silo (5 kg) for 50 (E50D) and 100 (E100D) days in triplicates. The E50D silages had higher in vitro digestibilities of DM (IVDMD, p<0.001) and neutral detergent fiber (IVNDFD, p=0.013), and lactate (p=0.009), and acetate (p=0.011) than those of E100D, but lower pH, lactic acid bacteria (LAB), and yeast. By inoculant application, LCH had highest IVDMD and IVNDFD (p<0.05), while MIX had highest lactate and lowest pH (p<0.05). The CON and LCH in E50D had highest LAB and yeast (p<0.05), whereas LBB in E100D had lowest (p<0.05). Therefore, this study concluded that LCH application improved the nutrient digesbility (IVDMD and IVNDFD) of lignified rye silage, and longer ensiling period for 100 days enhanced the fermentation characteristics of silage compared to ensiling for 50 days.

Recently, unmanned aerial vehicles (UAV, Drone) are highly regarded for their potential in the agricultural field, and research and development are actively conducted for various purposes. Therefore, in this study, to present a framework for tracking research trends in UAV use in the agricultural field, we secured a keyword search strategy and analyzed social network, a methodology used to analyze recent research trends or technological trends as an analysis model applied. This study consists of three stages. As a first step in data acquisition, search terms and search formulas were developed for experts in accordance with the Keyword Search Strategy. Data collection was conducted based on completed search terms and search expressions. As a second step, frequency analysis was conducted by country, academic field, and journal based on the number of thesis presentations. Finally, social network analysis was performed. The analysis used the open source programming language 'Python'. Thanks to the efficiency and convenience of unmanned aerial vehicles, this field is growing rapidly and China and the United States are leading global research. Korea ranked 18th, and bold investment in this field is needed to advance agriculture. The results of this study's analysis could be used as important information in government policy making.

DNA 바코드를 기반으로 하여 외국발 국내 입항 선박에서 검출되는 편승자 해충(hitchhiker insect pests)에 대한 모니터링을 수행하였다. 국내 입항 선박 조사는 2018년 6월 1일부터 2018년 9월 17일까지 약 109일간 111개 선박에 대해 실시하였다. 모니터링 대상 해충은 ‘보고잡기법(simply collecting method by hands)’으로 확보하였으며, 총 336개체 에 대해 종동정을 실시하였다. 그 결과, 확보 해충 중 농림축산검역본부에 ‘관리해충(Regulated insect pest)’으로 등재되어 있는 Noctua pronuba를 포함하여 총 13종 21개체(Amata formosae (Erebidae, Lepidoptera, 1개체), Arippara disticha (Pyralidae, Lepidoptera, 1개체), Chondracris rosea (Acrididae, Orthoptera, 4개체), Cyrtacanthacris tatarica (Acrididae, Orthoptera, 1개체), Euhampsonia serratifera (Notodontidae, Lepidoptera, 3개체), Lemyra rhodophilodes (Erebidae, Lepidoptera, 2개체), Lymantria xylina (Erebidae, Lepidoptera, 1개체), Malacosoma dentata (Lasiocampidae, Lepidoptera, 3개체), Neochauliodes meridionalis (Corydalidae, Megaloptera, 1개체), Noctua pronuba (Noctuidae, Lepidoptera, 1개체), Parasa pastoralis (Limacodidae, Lepidoptera, 1개체), Psilogramma lukhtanovi (Sphingidae, Lepidoptera, 1개체), Syntypistis viridipicta (Notodontidae, Lepidoptera, 1개체))의 국내 미서식종이 채집되었으며, 확보된 21개체 중 15개체는 살아있는 상태로 검출되었다. 특히, 관리해충인 N. pronuba의 경우, 싱가포르에서 광양을 거쳐 포항항으로 입항한 선박에서 검출되었는데, 이 종은 유럽을 비롯하여 중동 및 중앙아시아까지 서식하는 종으로 1990년대 북미 대륙에 침입한 것으로 확인된 바 있다. 이에 따라, N. pronuba를 비롯하여 현 조사에 확인된 국내 미서식 편승자 해충의 편승 유무에 대한 정밀 모니터링 뿐만 아니라 위 종들에 대한 체계적인 위험성 평가 역시 필요할 것으로 사료된다.

Cryopreservation of bovine embryos is used to efficiently implant surrogate mothers. It has been widely accepted that high lipid content in the oocyte interrupts its survival during freeze-thaw cycles. Serum component in the culture medium is thought to increase the embryo`s lipid contents. Conversely, L-carnitine stimulates lipid metabolism by transporting long chain fatty acids into the mitochondria. Objective of this study was to analyze the effect of L-carnitine supplementation in IVM medium and defined IVC medium on the development, lipid contents and the cryosurvival of bovine IVF embryos. 0.0, 1.5, 3.0 and 6.0 mM L-carnitine was supplemented in IVM medium, respectively (IVM-LC 0.0, LC 1.5, LC 3.0 and LC 6.0). Development rate from the 2cell to the morula stages was higher in IVM-LC 3.0 groups than those of IVM-LC 6.0 (p<0.05). But there were no significant differences among the other groups in the blastocyst rates and lipid content results. When 0.0, 1.5, 3.0 and 6.0 mM L-carnitine were supplemented in IVC medium (IVC-LC 0.0, LC 1.5, LC 3.0 and LC 6.0), development competence was not significantly different between those embryos. Lipid contents of embryos treated L-carnitine (IVC-LC 1.5, 3.0 and 6.0) were significantly lower than embryos of non-treated group. L-carnitine was supplemented 0.0, 1.5, 3.0, 6.0 mM during IVM and 3.0 mM during IVC (LC 0.0 - 3.0, LC 1.5 – 3.0, LC 3.0 – 3.0, LC 6.0 – 3.0) and cryosurvival of blastocysts confirmed after freezing-thawing. There were no significant differences on development, but LC 3.0 – 3.0 was significantly lower lipid contents than other groups. And LC 3.0 – 3.0 had better survival rates and hatched rates of blastocysts than LC 0.0 – 0.0. In conclusion, supplementation of L-carnitine in defined IVC medium decreases lipid contents. And L-carnitine supplementation improves cryosurvival and developmental ability of bovine IVF embryos.

The early-onset familial Alzheimer's disease (EOFAD/ FAD), the less common type of Alzheimer's disease (AD) currently affects a vast number of individuals worldwide. This type is being inherited as an autosomal dominant fashion. Missense mutations on Amyloid precursor protein (APP) and Presenilins 1 and 2 (PSEN1 & PSEN2) are known as major genetic factors in FAD. Conversely, missense mutations on microtubule-associated protein tau (MAPT) are also thought to involve. Up to date, several triple-transgenic animal models with muted forms of the human APP, PSENs and MAPT have been reported. Compared to other animals, canines are more emotional and their disease signs can be easily diagnosed. This attempt was to develop a triple transgenic canine model for the AD. We have obtained the coding sequences of APP, PSEN1 and MAPT from Dana-Farber/Harvard Cancer Center DNA resource core at HMS and incorporated several common AD mutations. The transgenic construct is composed of hNSE (ENO2) promoter-driven three AD genes fused together with modified 2A sequences. It was transfected into the canine fetal fibroblasts which were then used to perform somatic cell nuclear transfer (SCNT). The viable transgenic embryos were obtained after in vitro culture and the GFP was detected. In this study, we have successfully produced viable triple transgenic canine cloned embryos using SCNT technique. These transgenic canine embryos will be further developed into canines with FAD. The transgenic canines will be a good candidate in the AD research field.

This study was conducted to examine the effect of new inoculants on in vitro digestibility and fermentation characteristics of high moisture rye silage. Rye was harvested at heading stage and divided into 5 treatments, following: No additives(CON); L. plantarum R48-27(NI1); L. buchneri R4-26(NI2); mixture of NI1 and NI2 at 1:1 ratio(MIX); and L. buchneri(LB). The rye forage was ensiled into 10 L bucket silo for 100 days. In vitro digestibility of dry matter and neutral detergent fiber were highest(p<0.05) in NI2 silage. The pH in NI2 and LB silages were lower(p<0.05) than CON silage. Lactate concentration was highest(p<0.05) in NI1 silage. While concentrations of acetate and propionate were highest(p<0.05) in MIX silage. Lactates : acetate ratio was highest(p<0.05) in NI1 silage, but lowest in LB silage. Butyrate concentrations of NI2 and LB silages were lower(p<0.05) than that in CON and NI1 silages. Lactic acid bacteria (LAB) count in all inoculated silages was higher(p<0.05) than that in CON silage, while yeast count in LB silage was lower than in CON, NI1, and MIX silages. In conclusion, application of NI2 inoculant could improve potentially fermentation quality and digestibility of high moisture rye silage.

This study was carried out to estimate the effect of selected inoculants on chemical compositions and fermentation characteristics of rye silage. Rye was harvested at dough stage and divided into 5 treatments, following: No additives (CON); L. plantarum R48-27 (LP27); L. buchneri R4-26 (LB26); Mixture of LP27 and LB26 at 1:1 ratio (MIX); and L. buchneri (LB). The rye forage was ensiled into 10 L bucket silo for 100 days. The contents of NDF and ADF were lowest (P<0.05) in LB26. The pH in LB26, MIX, and LB were lower (P<0.05) than CON and LP27. Lactate content in LB was higher (P<0.05) than the others, while acetate content in LB26 and LB were higher (P<0.05) than that in CON and LP27. Lactate to acetate ratio was highest (P<0.05) in LB, but lowest in LB26. Lactic acid bacteria (LAB) count in LB was higher (P<0.05) than that in CON, while yeast count in CON was lower than in all silages applied inoculants. In conclusion, silages inoculated with LB26 could improve potentially the aerobic stability caused by increases of acetate and propionate concentrations.

The aim of this study was to evaluate the role of demineralized and particulate autogenous tooth, and interleukin-6 in bone regeneration. A demineralized and particulate autogenous tooth was prepared and human osteoblast-like cells (MG63) and human osteosarcoma cells were inoculated into the culture. The rate of cell adhesion, proliferation and mineralization were examined, and the appearance of cellular attachment was observed. An 8 mm critical size defect was created in the cranium of rabbits. Nine rabbits were divided into three groups including: An experimental group A (3 rabbits), in which a demineralised and particulate autogenous tooth was grafted; an experimental group B (3 rabbits), in which a demineralized, particulate autogenous tooth was grafted in addition to interleukin-6 (20 ng/mL); and a control group. The rabbits were sacrificed at 1, 2, 4 and 6 weeks for histopathological examination with H-E and Masson’s Trichrome, and immunohistochemistry with osteocalcin. The cell-based assay showed a higher rate of cell adhesion, mineralization and cellular attachment in the experimental group A compared with the control group. The animal study revealed an increased number of osteoclasts, newly formed and mature bones in the experimental group A compared with the control group. Eventually, a higher number of osteoclasts were observed in the experimental group B. However, the emergence of newly formed and mature bone was lower than in the experimental group A. The current results suggest that treatment with demineralized and particulate autogenous tooth and interleukin-6 is not effective in stimulating bone regeneration during the bone grafting procedure.

This study was conducted to estimate the effect of home or hetero fermentative lactic acid bacteria(LAB) on chemical composition, fermentation quality, and aerobic stability of rye silage. Rye forage was harvested at dough stage(28.9% of dry matter), chopped to 3-5 cm length, and divided into 4 piles for different inoculations as treatment, following 1) No additives(CON); 2) Lactobacillus plantarum at rate of 1.5 x 105 cfu/g of fresh forage(LP); 3) L. buchneri at rate of 1.2 x 105 cfu/g of fresh forage(LB); and 4) Mixture of LP and LB at 1:1 ratio(MIX). Rye silage was ensiled into 20 L bucket silo in quadruplicate for 0, 1, 4, 7, and 100 day periods. After 100 days of ensiling, the silage treated with LB had lower acid detergent fiber content(p<0.05), but higher in vitro dry matter digestibility(p<0.05). The LB and MIX reduced (p<0.05) pH more rapidly than CON and LP across the ensiling days, but had no difference on 100 days. Silage treated LP had lowest(p<0.05) acetic acid, but highest(p<0.05) propionic acid. In contrast, LB treated silage had highest(p<0.05) acetic acid, but lowest(p<0.05) propionic acid with the absence of butyric acid. On microbial count, LP treated silage had lowest(p<0.05) LAB, yeast, and aerobic stability, whereas LB and MIX treated silages had highest(p<0.05). Mold was not detected across all silages. Therefore, it could be concluded that heterofermentative LAB solely or combo with homofermentative LAB might improve in vitro dry matter digestibility, fermentation characteristics, and aerobic stability of rye silage harvested at dough stage.

Oocyte is the central factor in the bi-directional communication axis in the ovarian follicles. It controls the cumulus or granulosa cells to perform functions which are beneficial for its own development via secreting paracrine growth factors, including GDF9 and BMP15. The aim of this study was to investigate whether the recombinant GDF9 and BMP15 are able to promote meiotic resumption and cumulus expansion of canine COCs during IVM, as well as to demonstrate the actions of GDF9 and BMP15 in regulating the expression of connexin transcripts in the ovarian granulosa cells. As results, GDF9 and BMP15 significantly improved the meiotic resumption rate and cumulus expansion by activating ERK1/2 signaling. Treatments with GDF9 significantly improved the expression of CyclinB1 but inhibited the expression of Cx43 transcripts. In addition, cumulus expansion genes (MAPK1, Ptgs2, Tnfaip6 and Ptx3) were differentially improved by GDF9 and BMP15. In the ovarian granulosa cells, GDF9 suppressed the expression of Cx43 transcripts by binding ALK4/5/7 receptors and activation Smad2/3 signaling, whereas, BMP15 stimulated the expression of Cx43 transcripts by binding ALK2/3/6 receptors and activating Smad1/5/8 signaling. In conclusion, by regulating functions of granulosa/cumulus cells, oocyte has the potential to enhance the growth and maturation of itself.

Somatic cell nuclear transfer (SCNT) is the technique which generates embryos by transferring diploid nucleus into an enucleated oocyte, it has produced specific animals successfully in a variety of species. However, the developmental capacity of SCNT embryos is still relatively lower than that of embryos produced in vivo. Oocyte is a kind of lipid rich cells, its quality limits the efficiency of embryo production. L-carnitine is a co-enzyme facilitating the transportation of long chain fatty acids across the inner mitochondria membrane where fatty acids are used for generating adenosine triphosphate (ATP) via beta-oxidation. It also has antioxidant actions which may protect mitochondrial membranes and DNA against damage induced by reactive oxygen species (ROS). Whether L-carnitine is functional in bovine SCNT embryos are unknown. Therefore, the objective of this study was to examine the effects of L-carnitine on oocyte maturation and developmental competence of subsequent SCNT embryos. L-carnitine was supplemented during IVM, then intracellular ROS and GSH levels, mitochondrial activity, gene expression of COCs were analyzed at the end of IVM. SCNT embryos were produced subsequently, apoptosis detection and gene expression evaluation were performed in blastocysts. In the results, treatments with 1.5 mM and 3 mM L-carnitine significantly improved maturation rates (P<0.05). Treatments with 3 mM L-carnitine effectively induced improvement in nuclear maturation, intracellular GSH levels and mitochondrial activity, as well as a reduction in intracellular ROS levels (P<0.05). mRNA levels of CPT1A, ACAA1, ACAA2, AREG, EREG, SOD1, GPX4, GLUT1 and CDC2 transcripts were effectively up-regulated by 3 mM L-carnitine treatments (P < 0.05). Similarly, 3mM L-carnitine induced an increase in blastocyst developmental rates and an improvement in blastocyst quality (P<0.05). Our study indicates that L-carnitine treatment during IVM improves oocyte nuclear maturation and subsequent SCNT embryo development.

The mesenchymal stem cells (MSC) has been investigated as a source of stem cell therapy to replace and treat damaged cells. Human endometrial epithelial and stromal cells was isolated from hysterectomy tissue and the direct evidence of stem/progenitor cells in the human endometrium was identified. Endometrium derived stem cells (EnMSCs) are known to have a high proliferative ability, genetic stability, lack of tumorigenicity and low immnunogenicity during long-term cultivation. Here, we aimed to identify MSC in canine endometrium and characterize its potential to differentiate into decidua cells. EnMSCs were isolated from thrown-away spayed uterus of adult canine depending on their estrus cycle, and identified by flow cytometry, immunocytochemistry and flow cytometry with MSC specific markers. We then characterized the ability of EnMSCs by the doubling-time analysis, colony-forming units and MSC differentiation assays. Isolated EnMSCs expressed stem cell specific genes (Sox2, Oct4, Nanog, MCAM, Endoglin, Susd2 and IGTB) and MSC surface markers (CD90, CD44 and CD117). EnMSCs are also differentiated into adipogenic, osteogenic and chondrogenic cells morphologically under modified conditions with the expression of lineage specific genetic markers. EnMSCs showed higher proliferation ability than canine amniotic fluid derived MSCs which were used as a positive control. EnMSCs were cultured at low density (10, 20, cells/cm2) and initiated to form small colonies of loosely-arranged cells and gradually formed large colonies of densely-packed cells which underwent self-renewal with high proliferative potential which is similar to the clonogenicity feature of human endometrium-derived stem cells. EnMSCs were then induced to differentiate into decidua cells with 0.5 mM dbcAMP. After 14 days, EnMSCs changed their morphology into the elongated and rounded shape. The induced decidual cells expressed PRL and IGFBP1 which are typically expressed in decidua cells. In conclusion, we successfully isolated and characterized MSC in the canine endometrium which differentiated into decidua cells. These results showed that endometrium may be a promising source of stem cells, and furthermore raise the possibility of canine EnMSCs as a novel hypothetical decidualisation model of infertility associated with decidualisation insufficiency and implantation failure.

Oocyte is the central factor in the bi-directional communication axis in the ovarian follicles. It controls the cumulus or granulosa cells to perform functions which are beneficial for its own development via secreting paracrine growth factors, including GDF9 and BMP15. The aim of this study was to investigate whether the recombinant GDF9 and BMP15 are able to promote meiotic resumption and cumulus expansion of canine COCs during IVM, as well as to demonstrate the actions of GDF9 and BMP15 in regulating the expression of connexin transcripts in the ovarian granulosa cells. As results, GDF9 and BMP15 significantly improved the meiotic resumption rate and cumulus expansion by activating ERK1/2 signaling. Treatments with GDF9 significantly improved the expression of CyclinB1 but inhibited the expression of Cx43 transcripts. In addition, cumulus expansion genes (MAPK1, Ptgs2, Tnfaip6 and Ptx3) were differentially improved by GDF9 and BMP15. In the ovarian granulosa cells, GDF9 suppressed the expression of Cx43 transcripts by binding ALK4/5/7 receptors and activation Smad2/3 signaling, whereas, BMP15 stimulated the expression of Cx43 transcripts by binding ALK2/3/6 receptors and activating Smad1/5/8 signaling. In conclusion, by regulating functions of granulosa/cumulus cells, oocyte has the potential to enhance the growth and maturation of itself.

The OPU technology has been largely used in order to enhance genetic improvement in domestic animals. This study demonstrated that OPU in Hanwoo can be used for the effective technique to improve the reproductive efficiency. This experiment showed the longest times of OPU ever carried out in Hanwoo. In this study four donors were selected from Hanwoo by using DNA extraction and SNP maker. Individual donors have genes which are CAPN1, CAST and FASN and that contain dominant position. CAPN1, CAST genes are often thought to be related with tenderness and FASN gene is thought to be associated with oleic acid which is identified a monounsaturated fatty acid found naturally in many plant and animal products. In experiment 1, OPU technique was used to evaluate the influence of the number of oocytes recovery rate per session. Totally 50 times OPU sessions were performed and oocytes recovery at every 10 times session was evaluated. In case of H4, the OPU session could be done around 30 times after her calving. Compared to the average number of oocytes recovery, H1 was more efficient than H3. Considering this results, the current study showed that animals have considerable individual variation in numbers of oocytes. In this study, the average recovery rate in Hanwoo is similar to the recovery rate of the Bos Taurus. Individual donors have no significant difference among group for recovery rate during 30 sessions. However, Thoese showed significant decrease in the number of oocytes recovery rate after 40 session of OPU treatment. Therefore we conclude that the Hanwoo donor is considered to be suitable for 30 times of OPU treatment. In experiment 2, OPU technique was used to evaluate the influence of the number of recovery rate monthly. The average number of oocytes recovery rate showed no significantly difference for five months. However, the number of oocytes recovery rate decreased significantly during three months after first five months experiments. In experiment 3, whether donors are parity or non-parity, the average number of oocytes recovery rate was checked. However, we could not find any significant result from this experiment. In experiment 4, the developmental rate of in vitro produced embryos from OPU was compared with that from slaughterhouse. From this, cleavage rate of oocytes from OPU is significantly less than that from slaughterhouse. In conclusion, this study included a thorough analysis of oocytes and embryo production by OPU from Hanwoo. OPU can be successfully performed under a continuous regime for 8 month in Hanwoo. The current study shows the clear proof that OPU could be adopted to produce oocytes and embryos of better quality as an advanced technique replacing the usage of MOET in Korea. Finally, elucidation the basis for numerous oocytes obtained from Hanwoo may contribute to a better understanding of reproductive physiology in cattle.

Somatic cell nuclear transfer (SCNT) has been considered for preserving genetically valuable or endangered animals. Sapsaree is a Natianal Monument in Korea to maintian a pure pedigree. The aim of this study was to produce azoospermia Sapsaree using SCNT and identify normal reproductive ability of cloned azoospermia Sapsaree. Ear skin biopsy was performed on a thirteen-year-old azoospermia Sapsaree and ear skin fibroblasts were isolated for SCNT as donor cells. The fibroblasts were injected into enucleated in vivo matured oocytes, the couplets were electrical fused by two pulse of direct current (55 V for 15 μs) using titanium and platinum fusion needle and activated by calcium ionophore. Cloned embryos were surgically transferred into oviducts of natuarally estrus cycle synchronized recipient dogs. The fusion rate of platinum needle was 70%, which was higher than those of titanium needle (64.1%). Developmental rate to the 8 cells and 10 cell stages was higher in platinum needle group (24% and 16%, respectively) than those of platinum needle group (14.8% and 3.1%, respectively). Total 35 SCNT embryos were transferred into oviducts of 3 recipient dogs and one recipient finally delivered a puppy by caesarean section. As results, this study demonstrated that platinum fusion needle could be successfully make the reconstructed embryos and improve the efficiency of canine SCNT. Cloning azoospermia Sapsaree may contribute to conserve genetically valuable and unique pedigree. And further study should be confirm whether cloned live dog is azoospermia.

Prolonged communication between oocytes and the surrounding somatic cells is one of the unique reproductive physiology in canine. Paracrine Kit ligand (KITL) signaling is a well-known communication between granulosa cells and the oocyte. KITL is a cytokine growth factor secreted by granulosa cells that signals via the c-kit receptor expressed by oocytes. Paracrine factors, including growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15), exert their effects by binding with the kinase receptors expressed on the granulosa cells. However, the regulations of GDF9 and BMP15 in the canine KITL expression are currently poorly understood. Therefore, we investigated the effects of GDF9 and BMP15 on the expression of KITL in canine ovarian granulosa cells in vitro. In Annexin V assay recombinant GDF9 and BMP15 did not induce apoptosis in the cultured ovarian granulosa cells. When treated, FSH significantly increased KITL expression, and hCG suppressed its expression. When both FSH and hCG were treated, the expression of KITL was affected by GDF9 and BMP15 in dose and time dependent manner in the luteal granulosa cells. GDF9 (10 ng/mL) significantly decreased KITL expression after12 h. BMP15 (10 ng/mL) significantly also decreased KITL expression after 24 h. Western blot and immunochemistry results indicate that GDF9 activated Smad2/3. After blocking ALK 4/5/7 receptors by SB, GDF9 failed to activate Smad2/3, also BMP15 did not activate Smad1/5/8 after blocking ALK 2/3/6 receptors by DM. So GDF9 exerts its effects via using ALK 4/5/7 receptors to activate SMAD2/3 signaling, and BMP15 binds ALK 2/3/6 receptors to activate SMAD1/5/8 signaling. The expression of KITL was not changed by SB or DM treatment. However, the effect of GDF9 and BMP15, which decreased the expression of KITL, was suppressed by SB or DM treatment. In conclusion, this study provides the first evidence that recombinant GDF9 and BMP15 decrease KITL expression in canine ovarian granulosa cells.

The cancer and Parkinson's disease associated protein DJ-1 is multifunctional protein that involves in diverse cellular process. DJ-1 protein has a cellular protective role and promoted cell survival under an oxidative stress. However, the cellular protective mechanism of DJ-1 is not fully understand, and we needs to be further study their functions in novel organisms. In the present study, we investigated the protective role of DJ-1 against induced oxidative stress in canine cell line. On the basis of these experiments, canine DJ-1 overexpressing and null cell lines were established. The stable overexpression and down regulation of DJ-1 efficiency confirmed by the western blot analysis. Subsequently, the DJ-1 gene transfected cell lines and control cells were subjected to induced the oxidative stress, and then cell viability, cell proliferation assay, cellular apoptosis detection analysis (Annexin V and TUNEL assay), intracellular ROS and mitochondrial activity were measured appropriately. The results showed that DJ-1 overexpressed cells were up-regulated cell viability under oxidative stress conditions induced by the rotenone and hydrogen peroxide (H2O2), whereas loss of DJ-1 cells were down-regulated the cell survival activity. Additionally, overexpression of DJ-1 cells increased cell resistance to oxidative stress and inhibited the elevation of cell death and cellular ROS induced apoptosis. Moreover, DJ-1 overexpressed cells was increased mitochondrial functions by using confocal microscopy with MitoTracker staining. On the contrary to this, DJ-1 null cells show defective cellular protection and mitochondria activity against oxidative stress conditions. Our data indicate that canine DJ-1 protein attenuates cellular apoptosis and ROS generation, enhances the cellular survival activity and promote mitochondrial function under the oxidative stress, likewise other mammalian cells. Importantly, DJ-1 overexpression may be an important part of a protective strategy as a sensor for oxidative stress.

To obtain information on the sanitary of indoor environment in greenhouses used for shiitake cultivation, bacteria associated with larvae and imagoes of Sciaridae flies, pest to shiitake mushroom, were isolated and identified. A total of 1,048 bacterial colonies were isolated from the flies’ larvae and 984 bacterial colonies were isolated from the flies’ imagoes. Based on molecular analysis of the 16S rDNA sequence, Achromobacter xylosoxidans and Tetrathiobacter kashmirensis of β-proteobacteria, Enterobacter asburiae and Raoultella ornithinolytica of γ -proteobacteria, Curtobacterium sp. and Microbacterium thalassium of Actinobacteria, and Penibacillus taichungensis of Firmicutes were identified from the colonies of the flies’ larvae. While, Bacillus megaterium, B. thuringiensis, Lysinbacillus sphaericus and L. fusifomis of Firmicutes, Microbacterium thalassium and Citricoccus parietis of Actinobacteria, and Enterobacter asburiae of γ-proteobacteria were identified from the flies’ imagoes. Some of the isolated bacterial species were known be human pathogens. Overall, the results of this study suggested that mushroom fly carrying human pathogenic bacteria is one of sources impact on the sanitary of indoor environment of greenhouses used for shiitake cultivation.