Lysophosphatidic acid (LPA) is a bioactive lipid messenger involved in the pathogenesis of chronic inflammation and various diseases. Recent studies have shown an association between periodontitis and neuroinflammatory diseases such as Alzheimer’s disease, stroke, and multiple sclerosis. However, the mechanistic relationship between periodontitis and neuroinflammatory diseases remains unclear. The current study found that lysophosphatidic acid receptors 1 (LPAR1) and 6 (LPAR6) exhibited increased expression in primary microglia and astrocytes. The primary astrocytes were then treated using medium conditioned to mimic periodontitis through addition of Porphyromonas gingivalis lipopolysaccharides, and an increased nitric oxide (NO) production was observed. Application of conditioned medium from human periodontal ligament stem cells with or without LPAR1 knockdown showed a decrease in the production of NO and expression of inducible nitric oxide synthase and interleukin 1 beta. These findings may contribute to our understanding of the mechanistic link between periodontitis and neuroinflammatory diseases.

Mesenchymal stem cells in the dental pulp exhibit a tendency for differentiation into various dental lineages and hold great potential as a major conduit for regenerative treatment in dentistry. Although they can be readily isolated from teeth, the exact characteristics of these stem cells have not been fully understood so far. When compared to twodimensional (2D) cultures, three-dimensional (3D) cultures have the advantage of enriching the stem cell population. Hence, 3D-organoid culture and 3D-sphere culture were applied to dental pulp cells in the current study. Although the establishment of the organoid culture proved unsuccessful, the 3D-sphere culture readily initiated the stable generation of cell aggregates, which continued to grow and could be passaged to the second round. Interestingly, a significant increase in SOX2 expression was detected in the 3D-spheroid culture compared to the 2D culture. These results indicate the enrichment of the stemness-high population in the 3D-sphere culture. Thus, 3D-sphere culture may act as a link between the conventional and 3D-organoid cultures and aid in understanding the characteristics of dental pulp stem cells.

본 연구는 원통형 종이포트를 활용한 토마토 육묘시, 염스트레스를 활용하여 고온기 도장 억제가능성을 검토하기 위하여 수행되었다. 시험구는 K2SO4, KCl과 KH2PO4을 각 5, 10 dS·m-1로 처리하였고, 또한, 토마토 모종에 고염도의 칼륨을 처리하여 수분 및 저온스트레스 환경에서의 적응성 및 생존성을 조사하였다. 조사결과, 처리 농도가 높아질수록 지상·지하부 건물중, 옆면적, 순동화율 (NAR)이 감소하고, 경경과 충실도는 증가하였다. 수분 스트레스 처리 이후, 대조구는 심한 위조현상을 보였지만, KCl처리구는 양호하였다. 상대수분함량은 대조구에서 23%, KCl처리구에서 8% 감소 하였다. 또한, 대조구에 비하여 KCl 처리구는 저장시(9, 12 및 15°C) 모종의 손상 비율이 낮았다. 이와 같은 결과로 보아, KCl과 같은 고농도의 칼륨 처리가 원통형 종이포트 토마토 육묘의 도장 억제에 효과적이며 환경 스트레스 내성을 향상시키는 것으로 판단된다.

Lysophosphatidic acid (LPA) is a lipid messenger mediated by G protein-coupled receptors (LPAR1-6). It is involved in the pathogenesis of certain chronic inflammatory and autoimmune diseases. In addition, it controls the self-renewal and differentiation of stem cells. Recent research has demonstrated the close relationship between periodontitis and various diseases in the human body. However, the precise role of LPA in the development of periodontitis has not been studied. We identified that LPAR1 was highly expressed in human periodontal ligament stem cells (PDLSCs). In periodontitis-mimicking conditions with Porphyromonas gingivalis -derived lipopolysaccharide (Pg-LPS) treatment, PDLSCs exhibited a considerable reduction in the cellular viability and osteogenic differentiation potential, in addition to an increase in the inflammatory responses including tumor necrosis factor-α and interleukin-1β expression and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) activation. Of the various LPAR antagonists, pre-treatment with AM095, an LPAR1 inhibitor, showed a positive effect on the restoration of cellular viability and osteogenic differentiation, accompanied by a decrease in NF-κB signaling, and action against Pg-LPS. These findings suggest that the modulation of LPAR1 activity will assist in checking the progression of periodontitis and in its treatment.

Clear cell odontogenic carcinoma (CCOC), a very rare neoplasm located mostly in the mandible, has been regarded as a benign tumor. However, due to the accumulation of case reports, CCOC has been reclassified as a malignant entity by the World Health Organization. Patients with CCOC present with regional swelling and periodontal indications with variable pain, often remaining misdiagnosed for a long period. CCOC has slow growth but aggressive behavior, requiring radical resection. Histologic analysis revealed the monophasic, biphasic, and ameloblastic types of CCOC with clear cells and a mixed combination of polygonal and palisading cells. At the molecular level, CCOC shows the expression of cytokeratin and epithelial membrane antigen, along with markers that assign CCOC to the sarcoma family. At the genetic level, Ewing sarcoma breakpoint region 1-activating transcription factor 1 fusion is regarded as the key feature for identification. Nevertheless, the scarcity of cases and dependence on histological data delay the development of an efficient therapy. Regarding the high recurrence rate and the potential of distant metastasis, further characterization of CCOC is necessary for an early and accurate diagnosis.

The global mushroom industry has grown rapidly in recent years in terms of beneficial effects, market value, and demand. India has a wide range of agro-climatic conditions and is largely an agricultural country with a cultivated area of about 4.37 %, generating about 620 million tons of agro waste annually. Mushroom cultivation not only helps recycle agro wastes, but also fills the nutritional gap prevalent among a large population of India. Recently, government industrial policy and creative innovation has promoted research and other endeavors aiming towards the cultivation of mushrooms. Mushroom cultivation in India was initiated in Solan, in the mid-sixties. Mushroom cultivation has been successful in temperate regions of the Himalayas, the Western Ghats, and the hills of northeast India. Recently, many unemployed people have begun to adopt mushroom cultivation as a means of self-employment. It is high time that Indian mushroom cultivators and consumers became aware of the nutritional and medicinal values of cultivated and wild species of mushrooms. The total mushroom production in India between 2010 and 2017 was approximately 0.13 million tons, accounting for a 4.3% increase in the average growth rate of mushrooms per annum. In particular, the total production of white button mushrooms is the highest, with a share of about 73% of total mushroom production. In this review article, we have analyzed the current scenario of the Indian mushroom industry and its contribution to the economic growth of the country.

Xyrosaris Meyrick, 1907 is a small genus of the family Yponomeutidae, established by Meyrick (1907) for its type species Xyrosaris dyropa Meyrick, 1907. A total of nine species were recognized in the world. Most of species of the genus have been described by Meyrick at the early 20th century. Up to date, X. lichneuta Meyrick, 1918 has been reported from Korea. In the present study, we review the genus Xyrosaris from Korea, with two new species: X. sp.1 and X. sp.2. Key to species, illustrations of adult, male and female genitalia, diagnosis and short description are provided.

The genus Meganola (Nolidae, Nolinae) was established by Dyar (1898), with the type species Meganola conspicua Dyar, 1898, from America (Type locality: Texas, Colorado, Arizona). This genus comprises 80 species described in the Palaearctic and Oriental region. In this study, genus Meganola Dyar is reviewed from Korea, with description of a new species. We redescribed 12 species and one new species. Illustrations of adults and genitalia of all Korean species are provided, with a key to the genus of Meganola based on the male genitalia.

The mesenchymal stem cells (MSCs) that reside in dental tissues hold a great potential for future applications in regenerative dentistry. In this study, we used human dental pulp cells, isolated from the molars (DPCs), in order to establish the organoid culture. DPCs were established after growing pulp cells in an MSC expansion media (MSC-EM). DPCs were subjected to organoid growth media (OGM) in comparison with human dental pulp stem cells (DPSCs). Inside the extracellular matrix in the OGM, the DPCs and DPSCs readily formed vessel-like structures, which were not observed in the MSC-EM. Immunocytochemistry analysis and flow cytometry analysis showed the elevated expression of CD31 in the DPCs and DPSCs cultured in the OGM. These results suggest endothelial cell-prone differentiation of the DPCs and DPSCs in organoid culture condition.

Recently, we published a microinjection method for generating transgenic cattle using the DNA transposon system and their analysis by next-generation sequencing (Yum et al. Sci Rep. 2016 Jun 21;6:27185). In that study, we generated transgenic cattle using two different types of DNA transposon system, sleeping beauty (SB) and piggybac (PB), carrying Yellow fluorescent protein with SB (SB-YFP, female) and green fluorescent protein with PB (PB-GFP, male) under the control of the ubiquitous CAG promoter, respectively. The female and male founder cattle have been grown up to date (the female age: 40 months old, the male age: 33 months old) without any health issues. In genomic instability and blood analysis, there was no significant differences between wild type and founder cattle. In the present study, we confirmed germ-line transmission of the transposon-mediated transgene integrations and ubiquitous and persistent expression of transgene in second generation of offspring (F1). The F1 was born without any assistance and expressed GFP in the eyes without UV light. The ubiquitous expression of GFP was detected in skin fibroblast from the ear tissue and confirmed by genomic DNA PCR, which suggest that the transgene from the PB-GFP was successfully transmitted. Unfortunately, no transgene from SB-YFP were identified. To confirm the transgene integration site, the genomic DNA from blood was extracted and performed next-generation sequencing (NGS). The GFP gene was integrated in chromosome 4 (two copies), and 6. As results, a total of two copies of paternal transgene transmitted into the F1. All the integrated position was not related with coding region and there was no significant difference in genomic variants between transgenic and non-transgenic cattle. To our knowledge, this is the first report of germ-line transmission through non-viral transgenic founder cattle. Those transgenic cattle will be valuable resource to many fields of biomedical research and agricultural science.

Lycorine, a natural alkaloid extracted from the Amaryllidaceae plant family, was reported to various physiological and pharmacological effects including anti-cancer activity. Nevertheless, there is no report of the anticancer effect of lycorine in oral cancer cells. The effects of lycorine on cell proliferation and apoptosis were examined through trypan blue exclusion assay, 4’-6-diamidino-2-phenylindole (DAPI) stain, Live/Dead assay, Western blot analysis and RT-PCR. Lycorine suppressed cell viability and induced apoptosis in MC3 and HSC-3 cell lines. Lycorine decreased survivin protein but did not affect its mRNA. It regulated survivin through accelerating protein degradation in a time-dependent manner although neither proteasome nor lysosome was not associated with lycorine-mediated protein degradation. Collectively, our results suggest that lycorine may be a potential therapeutic anti-cancer drug candidate for the treatment of human oral cancer.

CRISPR/Cas9-induced knock-out/-in can be occurred at specific locus in the genome by non-homologous end joining (NHEJ) or homology directed repair (HDR). Here, we demonstrate the targeted insertion into the specific loci of embryo fertilized by semen from transgenic cattle via CRISPR/Cas9 system. Recently, we published on the efficient generation of transgenic cattle using the DNA transposon system (Yum et al. Sci Rep. 2016 Jun 21;6:27185). In the study, eight transgenic cattle were born following transposon-mediated gene delivery system (Sleeping Beauty and Piggybac transposon system) via microinjection. In the analysis of their genome stability using next-generation sequencing, there was no significant difference in the number of genetic variants between transgenic and non-transgenic cattle. All the transgenic cattle have grown up to date (the oldest age: 33 months old, the youngest age: 15 months old) without any health issue. One of transgenic male cattle expressing GFP reached puberty and semen was collected. Over 200 frozen semen straws were produced and some were used for in vitro fertilization (IVF). On seven days after IVF, expression of GFP was observed at blastocyst stage and was seen in 80% of the embryos. Another application is to edit the GFP locus of the transgenic cattle because long-term and ubiquitous expression of transgene didn’t affect their health. In one cell stage embryos produced using GFP frozen-thawed semen, microinjection of sgRNA for GFP, Cas9, together with donor DNA that included RFP and homology arms to link the double-strand break of sgRNA target site into fertilized eggs resulted in expression of RFP. This indicated that the GFP locus of transgenic cattle shows potential candidates for stable insertion of the functional transgene. Knock-out/-in for editing GFP locus using CRISPR-Cas9 might be a valuable approach for the next generation of transgenic models by microinjection. In conclusion, we demonstrated P-112 that transgenic cattle via transposon system are healthy to date and germ-line competence was confirmed. The GFP locus will be used as the potential target site for future gene engineering via genome-editing technology. Finally, all those animals could be a valuable agricultural and veterinary science resource for studying the effects of gene manipulation on biomedical research and medicine. This work was supported by BK21 PLUS Program for Creative Veterinary Science and Seoul Milk Coop (SNU 550-20160004).

The CRISPR/Cas9 system is proved to be a powerful tool for knock-out and knock-in in various species. By introducing genetic materials of two components (Cas9 and small guide (sg) RNA) into cells or pronuclear of the fertilized embryo, gene editing occurs. Some studies reported that efficiency of gene editing would be increased as Cas9 was integrated into cells or animals since Cas9 is indispensable in the CRISPR/Cas9 system. Accordingly, the production of Cas9 expressing cattle may provide the broadly used gene editing platform in cattle. For this study, Cas9 and RFP genes were cloned into PiggyBac (PB) transposon system. PB-Cas9-RFP and transposase were microinjected into 1436 in vitro fertilized embryos and 241 blastocysts were formed. Blastocysts with RFP expression accounting for 14.1% of total formed blastocysts were selected and transferred into 5 recipient cow. After gestation periods, four transgenic cattle were delivered without any veterinary assistance. From a transgenic cattle, ear skin tissue was collected for primary culture. On those primary cells, sgRNAs in DNA form for various genes such as PRNP, RB1 and BLG were transfected as 2ug of sgRNA per 5x105 cells using Nucleo factor system (Neon®, invitrogen, program#16). As expected, every group of each sgRNA delivered was confirmed to be mutated by T7E1assay. Those data demonstrated that for the first time, transgenic cattle with Cas9 expression were born, grown up to date and will be avaluable resource for genome-editing in cattle. This work was supported by BK21PLUS Program for Creative Veterinary Science and Seoul Milk Coop (SNU550-20160004).

This study investigated the antibacterial effects of Galla rhois extract (GRE) against Campylobacter jejuni and Campylobacter coli. The minimum inhibitory concentrations (MICs) of GRE against C. jejuni and C. coli were 0.28 and 0.55 mg/mL, respectively, and the corresponding minimum bactericidal concentrations (MBCs) were 4.4 and 5.5 mg/mL. C. jejuni treated with the MIC, MBC or 2×MBC of GRE showed significant inhibition of growth compared with that of the control group during the incubation period, and no viable bacteria were detected at 24 h after incubation. C. coli treated with MIC, MBC or 2×MBC of GRE also showed inhibition of growth compared with that of the control group during the incubation period, and in the C. coli cultures treated with MBC and 2×MBC of GRE, no viable bacteria were detected at 24 h after incubation. In conclusion, GRE is a candidate antibacterial agent against C. jejuni and C. coli, and may have applications for the control of Campylobacter infection in poultry.

본 연구는 오배자 에탄올 추출물 (GRE), 염소산나트륨 (SC) 그리고 오배자 에탄올 추출물과 염소산나트륨 합제(GS)의 B. abortus에 대한 항균효과를 확인하기 위해 수행 되었다. GRE, SC 그리고 GS를 B. abortus에 처리하여 배양한 후, B. abortus의 생존수를 확인하였으며, 마우스 탐식세포 내 감염된 B. abortus의 증식 억제효과를 경시별 (2, 24, 48시간)로 조사하였다. GRE, SC 그리고 GS는 각각 400 μg/mL 이하, 15 mM 그리고 0.6GS (GS 1, GRE 1,000 μg/mL + SC 30 mM) 이하의 농도에서 세포독성을 나타나지 않았다. 모든 처리구에서 B. abortus의 생존율은 용량- 의존적으로 현저하게 감소하는 결과를 나타내었다. 또한, GRE (400 μg/mL), SC (15 mM) 그리고 0.5GS (GRE 500 μg/mL + SC 15 mM)를 처리한 세포에서 배양 48시간 후에, B. abortus의 증식이 통계적으로 유의성 있게 감소하였으며 (GRE, p < 0.01; SC and 0.5GS, p < 0.001), 특히, GS를 처리한 경우, B. abortus의 세포내 증식이 GRE와 SC의 상승작용에 의한 강력한 항균효과를 나타내었다. 결론적으로, GS는 B. abortus에 대한 항균물질로서 유용할 뿐만 아니라, 식육과 우유 위생 분야에 적용할 수 있을 것으로 생각된다.