Ovarian folliculogenesis and the production of fertilizable oocytes depend on gap junctional intercellular communication within both the developing and the mature follicle. Gap junctions connect oocytes with granulosa cells and granulosa cells with each other. Various nutritional bio-molecules are known to be transferred to the growing oocyte from the granulosa cells via gap junction. Signals that regulate meiotic maturation of fully-grown oocytes pass through the oocyte-granulosa cell gap junctions. Gap junctions also play a critical role in regulating uterine blood flow, contributing to the maternal recognition and also implantation during pregnancy. Due to the challenge of various stressors the in vitro embryo developmental potentials are still suboptimal compared to in vivo. To identify the molecular mechanism of these stressors and to improve the existing embryo developmental potentials, the singlet oxygens quencher lycopene was added to the culture media to counterbalance the oxidative damage caused by ROS. In this study, we have patterned connexin like Cx43, Cx37, Cx32 and Cx26 at protein and transcription level during follicular growth, atresia and blastocyst stage by using immunohistochemistry, conventional PCR and RT-qPCR. Lycopene (0.2 μM) significantly (P < 0.05) increased the gap junctional communication protein (connexin) expression of Cx43, Cx37, Cx32, Cx26 as compared to the control group at both transcription and translation level during follicular growth, atresia and blastocyst stage. Lycopene potentiates ovarian folliculogenesis, provides the production of fertilizable oocytes and improved embryo developmental capabilities by increasing gap junctional intercellular communication.

This study investigated the use of bovine serum albumin (BSA) as alternatives to fetal bovine serum (FBS) in in vitro maturation medium. The oocyte maturation, cumulus cell-oocyte gap junctional communication, and development of bovine embryos were determined by assessing their cell number, lipid content, mitochondrial activity, gene expression and cryo-tolerance. Oocytes were cultured in TCM-199 supplemented with 1 μg/ml estradiol-17ß, 10 μg/ml FSH, 10 ng/ml EGF, 0.6 mM cysteine, 0.2 mM sodium pyruvate and either 8% BSA (BSA group), 10% FBS (FBS group), or neither BSA nor FBS (TCM group), and followed by in vitro fertilization and the zygotes were cultured in SOF-BE1 medium. The differences in embryo development between experimental groups were analyzed by one-way ANOVA. We have shown that the percentages of embryos that underwent cleavage and formed a blastocyst were non significantly different among all experimental groups (37.4 ± 1.5% for FBS group vs. 31.1 ± 3.9% for BSA group and 34.5 ± 1.6% for TCM group, six replicates were performed). Furthermore, there was no significant difference between the percentage of MII oocyte between FBS (71.8 ± 1.9%) and BSA groups (69.3 ± 2.3%). However, culture of oocytes with FBS increased (P < 0.05) the cumulus cell expansion as well as expression of gape junction proteins, CX37 and CX43, at both transcriptional and translation levels. We also found that FBS significantly increased total cell number and decreased the apoptotic index in day-8 blastocyst comparing to BSA group. The beneficial effects of BSA on embryos were associated with significantly reduced intracellular lipid content and increased mitochondrial activity in both oocytes and blastocyst. Taken together, these data suggest that supplementation of maturation medium with BSA, as alternatives to FBS, can be used as defined medium that support consistently the development of IVP bovine embryos.

Mammalian fetal ovaries contains numerous primordial germ cells, however fewer ones can yield mature oocytes due to apoptosis and follicle atresia. Successful in vitro reconstitution of primordial germ cells has recently had a significant effect in the field of assisted reproductive technologies. However, the regulatory mechanisms underlying oogenesis remain unknown and recapitulation of oogenesis in vitro remains unachieved. Therefore, development of methods for obtaining mature oocytes by culturing the fetal ovaries in vitro could contribute to clarify these mechanisms. We adapt an in vitro system for culturing mouse fetal ovaries that support successful follicle assembly and improve oocyte growth and maturation. Ovarian tissues from 12.5 days postcoitum (dpc) fetal mice were cultured in vitro and the matured oocytes were differentiated from primordial germ cells after a 31 days culture period. Our results demonstrate that mouse fetal germ cells are able to form primordial follicles with artificial ovarian cells, and that oocytes within the growing follicles are able to mature normally in vitro. Taken together, this in vitro culture system is expected to aid in the development of new strategies to identify the reasons behind failure of follicle assembly and offer a platform for innovative research into preservation of female germ cells and conservation of endangered species.

The aim of this study was to investigate the role of Src homology 2-containing phosphotyrosine phosphatase SHP2 in intricate signaling network invoked by oocyte to achieve cytoplasmic maturation and also blastocyst development. Activation of SHP2 regulates multicellular differentiation, proliferation and survival through numerous signal pathways. The most prominent pathway is RAS/PI3K and p-AKT signaling cascade, as a result mitogenic effect become enhanced. Oocytes were cultured in cisplatin an anticancer drug, but selective activator of SHP2 and our grouping were SOF medium alone, SOF + EGF, SOF + CISPLATIN 0.3 μM, and SOF + EGF + CISPLATIN 0.3 μM. We evaluated that EGF neutralizes the apoptotic effect of cisplatin as well as maintain the high expression of SHP2, as a result blastocyst development become boosted up. We also found that inhibition of SHP2 with its specific inhibitor PHPS1 5 μM decreases the blastocyst development and neutralizes growth factors effect. The developmental ability and quality of bovine embryos were determined by assessing their cell number, gene expression, immunofluorescence, and immunoblot. The differences in embryo development between experimental groups were analyzed by one-way ANOVA. Our results show that SHP2 have significant effect on MAP kinase pathways which expand the cumulus cells during oocyte maturation and blastocyst development as compare to inhibition of SHP2 with PHPS1. SHP2 not only transduce the signaling of epidermal growth factor but it also has a role in signal transduction of FGF and IGF. The expression of ERK, PI3K/p-AKT and mTOR was increased with EGF, but with the treatment of SHP2 inhibitor the expression of these genes become drop done. So we can conclude from these results that SHP2 is important for oocyte maturation as well as for blastocyst development.

Previous studies have shown that kisspeptin (Kp-10) is expressed in mammalian ovaries; however, the expression and role of Kp-10 in bovine ovarian granulosa cells are still unclear. In this study, we assessed the expression of Kp-10 and its effects on the proliferation and apoptosis of bovine granulosa cells. Immunohistochemical analysis showed that Kp-10 was expressed in the cytoplasm of bovine ovarian granulosa cells. Moreover, MTT (3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl-2- H-tetrazolium bromide) assays showed that 100 nM Kp-10 significantly inhibited the viability of granulosa cells (P<0.01). Flow cytometry analysis showed that Kp-10 could significantly increase accumulation of cells in the G1 phase, decrease accumulation of cells in the S phase, and promote apoptosis in bovine granulosa cells (P<0.05). Additionally, Kp-10 decreased the mRNA levels of Bcl-2, an anti-apoptotic gene; increased the mRNA levels of caspase-3, a pro-apoptotic gene; and increased the mRNA levels of Fas and Fasl, two membrane surface molecule genes (P<0.05). Thus, our findings demonstrated for the first time that Kp-10 inhibited proliferation and promoted apoptosis in bovine ovarian granulosa cells. These findings provide insights into our understanding of the role of Kp-10 in mediating the proliferation of bovine granulosa cells.

Transplantation of stem cells, such as mesenchymal stem cells (MSCs), is a promising strategy for treating several types of intractable disorders. Mechanistically, it could not only replace damaged cells by direct contribution, but also establish an anti-inflammatory or immunomodulatory microenvironment. However, the cellular mechanisms underlying molecular and biological properties of stem cells during ex vivo expansion and also after transplantation in pathological environments remain largely elusive. We recently developed the cyanoacrylamide-based coumarin derivatives (Fluorescent real-time thiol tracer; FreSHtracer*) reversibly react with glutathione for monitoring of glutathione levels in living stem cells. These probes revealed that glutathione levels are heterogeneous among subcellular organelles and among individual cells and show dynamic changes and heterogeneity in repopulating stem cells depending on oxidative-stress or culture conditions. Importantly, a subpopulation of stem cells with high-glutathione levels exhibited increased self-renewal and migration activities in vitro and showed improved therapeutic efficiency in treating asthma. Furthermore, employing a novel combination of longitudinal intravital confocal fluorescence imaging and microcystoscopy in living animals, we investigated the distributions and properties of transplanted multipotent MSCs derived from human embryonic stem cells at single-cell resolution in real-time by performing confocal imaging of bladder tissues in a rat model of IC/BPS for up to 6 months post-transplantation. These novel real-time monitoring strategies demonstrate the novel molecular insight for maintaining stem cell functions and also enhance understanding of the in vivo behaviors of the engrafted stem cells, which is crucial to determine the efficacy and safety of stem cell-based therapies. This strategy may facilitate the translation of various stem cell-based approaches into clinical practice.

Bemisia tabaci (Gennadius) is an important polyphagous pest and transmits plant pathogenic virus to numerous agriculture crops. We compared the efficacy of Beauveria bassiana ARP14 strain with commercialized GHA strain against 2nd instar and 4th instar of B. tabaci. The nymphs were exposed in 1×108 conidia/mL by dipping method. There was no difference in mycosis rate between the two strains in both 2nd and 4th instar. After 72 h, mycosis rate of 2nd instar was 100.0% in B. bassiana ARP14 strain followed by 97.4% in B. bassiana GHA strain. In 4th instar, B. bassiana ARP14 strain caused 100.0% while B. bassiana GHA strain caused 93.3%.

South Korea has over 0.38 million of managed honey bee (Apis cerana) colonies before 2009 years ago, which produce the highest quantity of honey in the Korea; however, almost colony (90%) were collapsed by Korean Sacbrood Virus (KSBV) in South Korea. Korean Sacbrood Virus (KSBV) is the pathogen of A. cerana Sacbrood disease, which poses a serious threat to honeybee A. cerana, and tends to cause bee colony and even the whole apiary collapse. Colony collapse of A. cerana was first reported on the Pyeong-Chang of the South Korea in 2009. Several scientists and governments has been tried research for cure the sacbrood disease in A. cerana colony by medicines and management techniques. Unfortunately, The sacbrood disease dosen`t improve. So, we were developed a better breed of A. cerana for resistance of sacbrood virus by selection and then artificial insemination. A. cerana breeding technique was first successful applied with A. cerana in Korean. Queens was grafted from sacbrood resistance line and then it was growing in sacbrood disease colony that was survived 100%. Altogether selected 18 queens were artificially inseminated and 2,000 drones of A. cerana in Korea was used to evaluate amount of semen collection. We are select two scabrood resistance A. cerana line (R and H). R line be used for rearing the Queen. Drone was reared in H line colony. The RH hybrid were not infected sacbrood virus even spread sacbrood virus (2×106). RH colonies have very excellent hygienic behavior, brood, and sacbrood disease resistance activity.

기후변화는 전지구적 현상이며 온난화를 주패턴으로 하며 이상기상 현상의 빈도가 높아지는 등 기후예측불확실성이 증대로 이어지고 있다. 이에 따라 농업생태계의 기후변화에 대한 적응과 변화를 지표하기 위한 수단으로 말벌류 지표종을 선발하였다. 기후변화지표종 선발을 위한 지표의 객관성을 높이기 위해 지표 수준별로 개량화하였으며, 이를 통해 장수말벌, 등검은말벌, 털보말벌, 황말벌 4종을 지표종으로 선발하였다. 이 종의 장기 모니터링을 위한 무인모니터링 시스템을 개발하였다. 이를 통해 전국적 모니터링 네트워크 등의 구축이 가능할 것으로 판단된다.

Drosophila suzukii (Matsumura) is one of the most destructive pest of thin-skinned fruits such as blueberry, strawberry, raspberry, and cherry. We tested effect of fruit damage on the infestation of D. suzukii by using partially peeled blueberry fruits in assays of behavioral choice and development. In choice test, D. suzukii preferred partially peeled blueberry fruit than normal. But in development test, we did not found difference in adult emergence rate between normal and partially peeled blueberry fruit.

We evaluated three previously known repellents of Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), i.e., carvacrol, cis-jasmone, and methyl jasmonate for the attraction of commercialized predators, Orius laevigatus (Fieber) (Hemiptera: Anthocoridae), and native predator, O. minutus (L.), in Y-tube olfactometer. Higher proportion of O. laevigatus (mated unfed adult females) moved to the arm of methyl jasmonate or cis-jasmone than the arm of clean air. However, O. minutus did not show any significant preference to the chemicals tested. These results suggest that, among the tested chemicals, methyl jasmonate or cis-jasmone would be a good attractant for O. laevigatus.

Lead free (1-x)(0.675BiFeO3-0.325BaTiO3)- xLiTaO3 (BFBTLT, x = 0, 0.01, 0.02, and 0.03, with 0.6 mol% MnO2 and 0.4 mol% CuO) were prepared by a solid state reaction method, followed by air quenching and their crystalline phase, morphology, dielectric, ferroelectric and piezoelectric properties were explored. An X-ray diffraction study indicates that lithium (Li) and tantalum (Ta) were fully incorporated in the BFBT materials with the absence of any secondary phases. Dense ceramic samples (> 92 %) with a wide range of grain sizes from 3.70 μm to 1.82 μm were obtained in the selected compositions (0 ≤ x ≤ 0.03) of BFBTLT system. The maximum temperatures (Tmax) were mostly higher than 420 oC in the studied composition range. The maximum values of maximum polarization (Pmax ≈ 31.01 μC/cm2), remnant polarization (Prem ≈ 22.82 μC/cm2) and static piezoelectric constant (d33 ≈ 145 pC/N) were obtained at BFBT-0.01LT composition with 0.6 mol% MnO2 and 0.4 mol% CuO. This study demonstrates that the high Tmax and d33 for BFBTLT ceramics are favorable for industrial applications.

The ability to recover the nearly limitless supply of uranium contained within the world’s oceans would provide supply security to uranium based fuel cycles. Therefore, in addition to U.S. national laboratories conducting R&D on a system capable of harvesting seawater uranium, a number of collaborative university partners have developed alternative technologies to complement the national laboratory scheme. This works summarizes the systems analysis of such novel uranium recovery technologies along with their potential impacts on seawater uranium recovery. While implementation of some recent developments can reduce the cost of seawater uranium by up to 30%, other researchers have sought to address a weakness while maintaining cost competitiveness.