The present study was designed to determine the effect of barn or cycle of grazing on changes of biochemical metabolites in prepartum and changes of milk composition in postpartum of dairy cows. For this purpose, a total of sixteen 25 months old Holstein primiparous dairy cows were allocated in two groups (n=8) with an average body weight of 571.61 ± 35.30 kg (Barn) and 578.10 ± 39.20 kg (Grazing). The study was conducted from June 2018 to October 2018. Results revealed that barn raised dairy cows had a higher increase in their serum albumin and calcium level on day 14 prepartum. However, the level of palmitic acid, saturated fatty acid increased significantly, and the level of fat, oleic acid, γ-linoleic acid, arachidonic acid and unsaturated fatty acids decreased significantly in barn raised dairy cow’s milk on day 14 postpartum. There were no significant differences observed with respect to all other biochemical metabolites, fatty acids and minerals between barn raised and cycle grazing dairy cows during prepartum and postpartum. Our study results could serve to a better understanding of barn raised cow with respect to changes of biochemical metabolites in prepartum and changes of milk composition, fatty acids and minerals content in grazing dairy cows in postpartum for estimating their physiological status.

BALB/c mice were vaccinated with Brucella (B.) abortus recombinant protein L27 (50S ribosomal protein L27) cloned into a pMal vector system. L27 was induced, purified and injected intraperitoneally (IP). Mice were vaccinated on 0-, 15- and 35-day. Serum cytokines were evaluated on 36- and 49-day from first vaccination. Mice were intraperitoneally infected with 5×104 CFU of virulent B. abortus 544 on day-50 and sacrificed after two weeks from infection. Bacterial burden from the spleen was quantified and showed a 0.7- and 0.9-log reduction in vaccinated mice in comparison to PBS and MBP (maltose binding protein) groups respectively. Cytokines in the serum demonstrated increased interferon-gamma (IFN-γ) and other pro-inflammatory cytokines such as macrophage chemoattractant protein-1 (MCP-1) and interleukin 6 (IL-6). On the other hand, interleukin 10 (IL-10) was attenuated in the sera of vaccinated mice. This cytokine profile is indicative of a cell-mediated type of immune response which is favorable for the eradication of intracellular infections. The current study showed the potential of another B. abortus ribosomal protein in inducing protective immunity against B. abortus infection.

A simple and fast analytical method based on liquid chromatography-tandem mass spectrometry was developed for detection of the veterinary drugs acetanilide, anthranilic acid, antipyrine, cyproheptadine, diphenhydramine, DLmethylephedrine, and phenacetin in bovine milk. The target analytes were extracted from milk samples by using acetonitrile followed by clean-up with C18 and liquid-liquid purification with saturated n-hexane. A reverse-phase analytical column was employed with a mobile phase comprising (A) 0.1% formic acid in distilled water and (B) 0.1% formic acid in acetonitrile to achieve the best chromatographic separation. Matrix-matched calibration curves (r2 ≥ 0.9986) were constructed using six concentrations (1, 2, 5, 10, 20, and 40 μg/kg) of drugs in the milk matrix. Recoveries at three drug-spiking levels (5, 10, and 20 μg/kg) ranged from 71.2% to 103.8% with intra-day and inter-day relative standard deviation (RSD) values of ≤ 8.6%. The calculated limits of quantification (LOQ) were 0.19-7.1 μg/kg.

A previous studies depicting origin and sequence variability of the species using DNA barcoding region with the samples collected from Korea showed relatively low sequence variability. Thus, additional markers that reveal higher variability were necessitated to scrutinize population structure in connection with dispersal and invasive dynamics among international populations. Therefore, we sequenced two complete mitochondrial genomes (mitogenomes) of M. pruinosa from the two haplotypes occurring in Korea (H1 and H3). Comparison of the two mitogenomes each with 16,312 and 16,314 bp evidenced that one region located in the A+T-rich region to provide higher number of haplotypes (4 vs. 3), sequence divergence (1.636% vs. 0.636%), and variable sites (7 vs. 3) than those of DNA barcoding region from the screening test using 13 representative individuals. This variable region, in concatenation with the currently available DNA barcoding region might be useful for population genetic analysis of worldwide populations including those of Korea. †These authors contributed equally to this paper.

The morphological identification of alate aphids needs reliable and appropriate methods such as morphometrics. In previous studies, the seven species which aphids have been known as migrating to potato had been separated into genus levels using specific points of forewing vein (Landmarks). Adding the number of secondary sensillum of antennae, setae of abdomen, etc., we experimented to separate four species of genus Aphis (Aphis craccivora, Aphis fabae, Aphis gossypii, Aphis spiraecola). After taking each of 25X twenty photographies of four species, we calculated and measured coordinates of landmarks, the number of secondary sensillum of antennae, dorsal setae of 8th abdominal segment, anterior/posterior genital plate setae and cauda, and then performed the principal component analysis using PAST3 and the canonical variate analysis using MORPHO J, only with a landmarks. As results, in PCA, Aphis gossypii has been separated, but the others are overlapping. In CVA, all species of aphis has been separated.

최근 파밤나방 등 다양한 나비목 해충에서 diamide 계통의 살충제 (IRAC group 28)의 저항성이 많이 보고가 되고 있다. 국내에서도 이미 배추좀나방과 파밤나방의 저항성이 보고되었는데, 본 연구에서 사용된 강릉에서 2018년 채집한 파밤나방의 경우 Chlorantraniliprole에 대하여 감수성 대비 2,500배 이상의 높은 저항성을 나타냈다. 추천 농도 처리 시 30% 이하의 방제가를 나타냈으며 Cyantraniliprole, Cyclaniliprole, Flubendiamide에서 높은 교차저항성을 나타냈다. 이 저항성 집단에서 diamide 계통 살충제의 작용점인 ryanodine receptor에 이미 보고된 G4946E 돌연변이는 발견되지 않았으나 새로운 I4760M 돌연변이를 발견하였는데, 2014년 배추좀나방에서 보고된 돌연변이와 위치가 동일하였다. 따라서 I4760M 돌연변이는 파밤나방에서 diamide 저항성에 관여 할 것으로 판단한다.

목적 : 본 연구는 신체적 활동이 경도인지장애 노인의 실행기능에 미치는 효과의 근거를 제공하고자 한다. 연구방법 : Preferred Reporting Items for Systematic Reviews and Meta-Analysis(PRISMA) guidelines의 보고 지침 체크리스트 항목을 바탕으로 체계적 고찰방법을 사용하였다. 전자 데이터베이스인 PubMed, CINAHLPychINFO, Embase, Cochrane를 사용하여 자료를 수집하고, Cochrane Risk of Bias Tool을 통하여 각 연구의 비뚤림 위험을 평가하였다. 검색어에는 “MCI” OR “mild cognitive impairment” AND “physical activit*” OR “physical performance” OR “physical exercise*”, AND “executive function*”을 사용하여 최종 2005년부터 2017년까지 무작위대조군 연구 11편을 선정하였다. 결과 : 중재는 주로 그룹으로 진행 되었으며 복합적 중재보다는 단일 중재가 많았다. 중재를 진행한 전문가는 피트니스 전문가, 물리치료사, 작업치료사가 많았다. 실행기능을 측정한 도구로는 TMT-A(21.4%)와 TMT-B(17.9%)가 가장 많았으며, 중재로 사용된 신체적 활동으로는 유산소 운동과 (8편, 72.7%) 무산소 운동 (7편, 63.6%)이 많았다. 유산소 및 무산소 운동을 포함한 중재는 실행기능 향상에 유의한 효과가 있었으나, 여가 활동을 제공한 중재는 실행기능에 유의한 변화가 없었다. 결론 : 본 연구를 통해 신체적 활동이 경도인지장애 노인의 실행기능에 효과를 확인할 수 있었다. 작업치료는 경도인지장애노인의 실행기능 향상을 위한 중재로 인지적 접근과 더불어 신체적 활동에 관한 다양한 연구들이 활발하게 이루어 져야 할 것이다.

In this study, we examined the protective immunity of a combination of seven Brucella abortus recombinant proteins; superoxide dismutase (rSodC), riboflavin synthase subunit beta (rRibH), 50S ribosomal protein (50s rL7/L12), nucleoside diphosphate kinase (rNdk), malate dehydrogenase (rMDH), arginase (rRocF), and elongation factor (rTsf) cloned in a pMal vector system and expressed in DH5α. Mice groups were immunized thrice with a combined subunit vaccine (CSV-7) at 0, 2, and 5 weeks and subsequently challenged with B. abortus at 5 × 104 CFU at 6 weeks. At four weeks post-infection, the mice were sacrificed and the bacterial burden in their spleens was quantified. Results revealed bacterial log reductions of 0.63 and 0.34 in comparison to PBS and maltose-binding protein (MBP), respectively. Cytokine profiling revealed a marked increase in IFN-γ (interferon-gamma), MCP-1 (macrophage chemoattractant protein-1) and IL-6 (interleukin 6) cytokines at 5-weeks post-immunization. On the other hand, only TNF was heightened at 7-weeks post-immunization. In general, this cytokine profile is consistently reflective of a Th1 immune response, which is beneficial for host immunoresistance.

Exosomes are Nano-sized lipid vesicles secreted from mammalian cells containing diverse cellular materials such as proteins, lipids, and nucleotides. Multiple lines of evidence indicate that in saliva, exosomes and their contents such as microRNAs (miRNAs) mediate numerous cellular responses upon delivery to recipient cells. The objective of this study was to characterize the different expression profile of exosomal miRNAs in saliva samples, periodically isolated from a single periodontitis patient. Unstimulated saliva was collected from a single patient over time periods for managing periodontitis. MicroRNAs extracted from each phase were investigated for the expression of exosomal miRNAs. Salivary exosomal miRNAs were analyzed using Affymetrix miRNA arrays and prediction of target genes and pathways for its different expression performed using DIANA-mirPath, a web-based, computational tool. Following the delivery of miRNA mimics (hsa-miR-4487, -4532, and -7108-5p) into human gingival fibroblasts, the expression of pro-inflammatory cytokines and activation of the MAPK pathway were evaluated through RT-PCR and western blotting. In each phase, 13 and 43 miRNAs were found to be differently expressed (|FC| ≥ 2). Among these, hsa-miR-4487 (|FC|=9.292005) and hasmiR- 4532 (|FC|=18.322697) were highly up-regulated in the clinically severe phase, whereas hsa-miR-7108-5p (|FC|= 12.20601) was strongly up-regulated in the clinically mild phase. In addition, the overexpression of miRNA mimics in human gingival fibroblasts resulted in a significant induction of IL-6 mRNA expression and p38 phosphorylation. The findings of this study established alterations in salivary exosomal miRNAs which are dependent on the severity of periodontitis and may act as potential candidates for the treatment of oral inflammatory diseases.

This study evaluated the protective effects of a combination of eight B. abortus recombinant proteins that were cloned and expressed into a pMal vector system and DH5α: nucleoside diphosphate kinase (rNdk), 50S ribosomal protein (rL7/L12), malate dehydrogenase (rMDH), DNA starvation/stationary phase protection protein (rDps), elongation factor (rTsf), arginase (rRocF), superoxide dismutase (rSodC), and riboflavin synthase subunit beta (rRibH). The proteins were induced, purified, and administered intraperitoneally into BALB/c mice. The mice were immunized three times at weeks 0, 2, and 5 and then infected intraperitoneally (IP) with 5×104 CFU of virulent B. abortus 544 one week after the last immunization. The spleens were collected and the bacterial burden was evaluated at four weeks post-infection. The results showed that this combination produced a significant reduction of the bacterial burden in the spleen with a log reduction of 1.01 compared to the PBS group. Cytokine analysis revealed induction of the cell-mediated immune response in that TNF (tumor necrosis factor) and proinflammatory cytokines IL-6 (Interleukin 6) and MCP-1 (macrophage chemoattractant protein-1) were elevated significantly. In summary, vaccination with a combination of eight different proteins induced a significant protective effect indicative of a cell mediated immune response.

The ovum pick up(OPU) technique can be used to produce embryos after in vitro culture of ovarian oocytes, can be used for early securement for effective herd early proliferation and excellent Hanwoo genetic resources, It is attracting attention as a very important technique for establishing technology. In addition to in vitro culture techniques, the number of oocytes retrieved depends on the environment and timing of the OPU. This study was conducted to compare and examine seasonal effect to the differences in the number of recovered oocytes, recovery rate and embryo development rate using Korean cattle kept in animal genetic resource research center by OPU technique. The grade of COCs was evaluated by microscopic examination. Grade A had 3 or more layers of cumulus cell and compact cytoplasm. Grade B had 1~3 layers of cumulus cell and compact cytoplasm. Grade C had 1 layers cumulus cell and compact cytoplasm. Grade D was denuded oocyte and poor cytoplasm. The recovery rate was 47.8±3.4% in summer (June to August) and 51.6±3.8% in autumn (September to October). The number of oocytes was 5.7±0.6 in summer and 5.2±0.7 in autumn. Oocyte grade A and B was 46.2%±6.3% in summer and 51.1±5.0% in autumn. The cleavage rate was 46.1±7.1% in summer and 65.0±11.3% in autumn. Blastocyst development rate was 19.9±9.4% in summer and 29.0±8.7% in autumn. There was no difference the recovery rate of oocytes and the number of embryos between summer and autumn. Cleavage rate and blastocyst rate of autumn was higher than summer. we will investigate to study the appropriate method for the production of Hanwoo embryos and the systematic comparison.

The present study was to assess the in vivo embryo production efficiency using the semen separated according to sex during superovulation in Hanwoo. Seventy Hanwoo donor cows were flushed on day 7 of estrus cycle with same FSH and artificial insemination by the same technicians. Embryos were recovered on 7 days after the third insemination by flushing the uterus with embryo collection medium. KPN semen straws used artificial insemination contained 20 million sperm (total number 60 million per donor). Sex-sorted semen straws contained 4 million sperm (total number 12 million per donor). The results obtained were as follows: No differences were observed in the efficiency of superovulation rates on KPN semen 87%, and sexed semen 100%, respectively. The mean numbers of total embryos are each 12.58 ± 8.31 and 13.25 ± 7.86. The mean numbers of transferable embryos, sexed semen were significantly lower than KPN semen (3.75 ± 1.98 vs. 8.23 ± 6.07, P<0.05). The rates of unfertilized embryos from superovulation using sexed semen were significantly higher than KPN semen (50% vs. 15%, P<0.05). The rate of degenerated 2-cell embryos from sexed and KPN semen was 60.87% and 11.11%, respectively (p<0.05). In conclusion, these results indicate that superovulation using sexed semen was useful, but efficient embryo production was important to reducing the damage caused by the Flowcytometer-based sperm sorting procedure.

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.

Epizootic HPAIV, H5N6, and H5N8 infections produced severe loss in poultry and wild birds in the Republic of Korea from 2016 to 2017. But pathological lesions and antigen distribution of the novel HPAIV H5N6 clade 2.3.4.4 in natural cases have been rarely reported. Herein, we describe the pathological lesions and antigen localization in chickens (layer and Korean native), ducks, and Japanese quail naturally infected by HPAIV H5N6. Grossly, severe reddening, swelling, and some necrotic foci, which were similar to septicemia or viremia, were observed in skin and many visceral organs including trachea, lung, liver, spleen, and pancreas. Histopathologically, pulmonary congestion and edema, as well as necrotizing hepatitis, splenitis, pancreatitis, myocarditis, and encephalitis were observed. Immunohistochemically, numerous HPAIV antigens were detected in necrotic parenchymal cells and in blood vessels of the respiratory, lymphoid, digestive, urinary, nervous, and cardiovascular systems. The results indicate that HPAIV H5N6 spread to the entire body via blood and caused severe damage throughout the entire body. The HPAI H5N6 clade 2.3.4.4 virus was isolated from samples of all four cases.