A pregnancy diagnosis is an important standard for control of livestock’s reproduction in paricular dairy cattle. High reproductive performance in dairy animals is a essential condition to realize of high life-time production. Pregnancy diagnosis is crucial to shortening the calving interval by enabling the farmer to identify open animals so as to treat or re-breed them at the earliest opportunity. MicroRNAs are short RNA molecules which are critically involved in regulating gene expression during both health and disease. This study is sought to establish the feasible of circulating miRNAs as biomarkers of early pregnancy in cattle. We applied Illumina small-RNA sequencing to profile miRNAs in plasma samples collected from 12 non-pregnant cows (“open” cows: samples were collected before insemination (non-pregnant state) and after pregnancy check at the indicated time points) on weeks 0, 4, 8, 12 and 16. Using small RNA sequencing we identified a total of 115 miRNAs that were differentially expressed weeks 16 relative to non-pregnancy (“open” cows). Weeks 8, 12 and 16 of pregnancy commonly showed a distinct increase in circulating levels of miR-221 and miR-320a. Through genome-wide analyses we have successfully profiled plasma miRNA populations associated with pregnancy in cattle. Their application in the field of reproductive biology has opened up opportunities for research communities to look for pregnancy biomarker molecules in dairy cattle.

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.

Oxidative stress is one of common cause of fatty changes in the liver. Antioxidant capacity was confirmed in various vegetables including black radish (Raphanus sativus L. var niger). Fermentation of vegetables using Lactobacillus plantarum has been known to generate bioactive components. This study was conducted to determine if fermented black radish (FBR) ameliorates oxidative liver injury induced by CCl4 in rats. To accomplish this, FBR (250 and 500 mg/ kg) was orally administered to rats for 7 consecutive days, single CCl4 (1.5 mL/kg) treatment or no treatment orally. Serum chemistry at 24 hours after CCl4 injury showed that FBR (500 mg/kg) significantly reduced the level of both alanine aminotransferase and aspartate aminotransferase in CCl4 exposed rats. Moreover, FBR treatment significantly increased radical-scavenging effects in livers with the reduction of lipid peroxidation in CCl4 exposed rats. Histopathologic findings including Kupffer cell activation in the liver of each group matched those of serum chemistry. Collectively, black radish, through fermentation, exerts hepatoprotective capacity in CCl4 induced liver injury in rats through anti-oxidation.

Most patients with chronic low back pain experience functional disability of trunk muscle, and limitations in physical activity. While there are many types of exercise programs available, in recent years sling exercise has been emerging as the exercise program for spinal stabilization. It has been supported by a great amount of research with positive findings on its effectiveness. This research studies the effects of bridging exercise, conducted on a sling, on pain level and trunk muscle activation in supine, sidelying, and prone positions during a 4 weeks period. 10 healthy people(normal group, n=10) and 28 patients with low back pain participated in this study. 28 patients were divided into two groups; one group participated in exercise with the sling(experimental group, n=14) and the other group exercised without the sling(control group, n=14). They were asked to use the Numerical Rating Scale(NRS) to answer to the level of their pain they felt (no pain: 0 point, severe pain: 10 points). During sling bridging exercises, the muscle activity level in each muscle measured in each position was standardized as three seconds of EMG signals during five seconds MVIC. In conclusion, the experimental group with four weeks of sling bridging exercise experienced a statistically significant reduction in the pain level(p<.05) and increase in the muscle activities of erector spinae when in supine position, internal oblique when in sidelying position, and rectus abdominis in prone position(p<.05). Regular sling bridging exercise reduces the low back pain and enhances other trunk muscle activation, thereby positively affect spinal stabilization.

In using both hands, everyone dominantly use one hand and it is called left-handedness or right-handedness person. Measurements of grip and pinch strength provide objective indexes to represent functional integrity of the upper extremity. This study was conducted for thirty female college students(19 right-handedness and 11 lefthandedness). For assessment of the type of handedness, questionnaire was used; for grip strength, Jamar dynamometer was used; for pinch strength, Jamar pinch gauge was used. In right handedness, the grip and pinch strength of the dominant right hand was significantly higher than those of the non-dominant hand. In addition, regular exercises were shown to give influences on reduction of strength gaps between dominant and non-dominant hands. In both groups of left and right handedness, the grip and pinch strength of the dominant hand were significantly higher than those of the non-dominant hand, and regular exercises were shown to give influences on reduction of strength gaps between dominant and non-dominant hand.

From a conversation-analytic perspective, this paper examines the ways in which the Korean sentence-ending suffix (SES) -ketun is used in naturally occurnng sp아<en Korean 이 scourse. Focusing on the question of how the practice of informing implemented by the ketun-utterance provides a basis for the sequentially implemented action of account-giving (Kim & Suh 2뼈)， it a n따yzes (i) the nature of the information marked by -ketun in terms of how it is grounded, (ii) the sequential contexts where the informing sequence that -ketun generates is embedded, and (iii) features of the ketun -marked account with reference to the practices of formulating non-negotiable upshot, evo싸 ng positionally relevant identities, unpackaging information geared to enlightening the address얹. and stance-shifting. These practices are analyzed as being constitutive of the action-organizational features of the ketun-marked account, whose upshot is proposed to be that of evo써 ng and managing the information gap or disparity in knowledge asseπed by the speal<er to exist between the partlclpants.

Background: Angelica gigas, A. sinensis, and A. acutiloba are commercially important in the herbal medicine market, and among them, A. gigas has the highest economic value and price. However, their similar morphological traits are often used for fraud. Despite their importance in herbal medicine, recognition of the differences between Angelica species is currently inadequate. Methods and Results: A multiplex polymerase chain reaction (PCR) method was developed for direct detection and identification of A. gigas, A. sinensis, and A. acutiloba. The gene for the distinction of species was targeted at ITS in the nucleus and trnC-petN gene in chloroplasts. The optimized multiplex PCR in the present study utilized each Angelica species-specific primer pairs. Each primer pair yielded products of 229 base pairs (bp) for A. gigas, 53 bp for A. sinensis, 170 bp for A. acutiloba. Additionally non-specific PCR products were not detected in similar species by species-specific primers. Conclusions: In the present study, a multiplex-PCR assay, successfully assessed the authenticity of Angelica species (A. gigas, A. sinensis, and A. acutiloba). and whole genome amplification (WGA) was performed after DNA extraction to identify, the species in the product. The detection method of raw materials developed in the present study could be applied to herbal medicine and health functional food management.

Survival of embryos largely depends on yolk processing during early development. Proteolytic enzymes, cathepsin B & D (ctsb & ctsd) are known to have some important roles in yolk processing of various fish species. Mature female red spotted groupers were injected with human chorionic gonadotropin (HCG) to induce ovulation. The fertilized eggs and embryos were sampled at 0, 4 and 24 HPF (hours post fertilization). Survivals of each groups of embryos were checked at 24 and 48 HPH (hours post hatching). Transcripts of ctsb & ctsd showed the highest level at 0 HPF and relatively high at 4 HPF, but greatly decreased at 24 HPF. In bad egg quality group (BE, embryos survived until 24 HPH), transcript level of ctsb at 4 HPF were significantly lower than the transcript level at the same stage in good egg quality group (GE, embryos survived until 48 HPH) while no significant change of ctsb transcript level was observed at 0 or 24 HPF between BE and GE. Transcript level of ctsd was decreased at 24 HPF, but the difference was not as strong as the case of ctsb transcript. These results suggest that maternal ctsb transcript rather than ctsd transcript is likely to be involved in egg quality resulting in the difference of survival rate of embryos at early developmental period in this species.

Red spotted grouper, Epinephelus akaara, is a popular aquaculture species and a protogynous hermaphrodite. Induction of artificial sex change at the time of primary sex differentiation is of interest but has not been successful due to the lack of necessary basic information. To find out the potential neuroendocrine influence on the primary sex differentiation, the expression of key genes in the brain was investigated during the formation of ovarian cavity. Expression of cyp19a1b, esr1, gnrhr1, fsh, lh and cga in the brain was positively associated with the formation of ovarian cavity, showing gradual increase as the formation proceeds. However, the expression of gnrh1 was suppressed during the early part of the ovarian cavity formation, signifying potential hypothalamic influence on the primary sex differentiation in this species.

Background : Despite the presence of various bioactive compounds in ginseng, there is lack of study on the variations of bioactive compounds in ginseng according to the cultivation of soil and the applied fertilizer types (or amount). Therefore, this study aims to examine the variations of 37 fatty acids (FA) and 8 vitamin E (Vit-E) vitamers in 6-year-old ginseng root cultivated in different soil types with different fertilizers regimes. Methods and Results : The profiling of 37 FAs and 8 Vit-E vitamers in 6-year-old ginseng roots was measured by gas chromatography coupled with a flame ionization detector, and then these results were statistically analyzed with chemometrics. The FA and Vit-E content in ginseng roots varied significantly with respect to soil cultivation conditions due to organic fertilizer types and amounts used. Unsaturated FA in ginseng is approximately 2.7 fold higher than the saturated FA. Linoleic, palmitic, and oleic acids were the most abundant FAs found in the ginseng roots. Also, the major Vit-E vitamer found in ginseng root is α -tocopherol. In particular, the application of rice straw compost or food waste fertilizer was increased to create nutritionally-desirable FAs and bioactive Vit-E in ginseng root. In addition, phytonutrient profiling coupled with chemometrics can be used to discriminate the cultivation conditions of ginseng. Conclusion : This preliminary study extends our understanding about the variations of FA and Vit-E in ginseng root depending on cultivation conditions. Hence, these results can be useful as basic information for reliable ginseng production containing high amounts of phytonutrients in a paddy-converted field.

Red spotted grouper, Epinephelus akaara is a popular aquaculture species in many Asian countries. This species is a protogynous hermaphrodite that first differentiates into female and changes to male later. Due to this reproductive characteristic, stable supply of male and female gametes is a key to the success of seed production in this species. Thus, understanding early gonadal differentiation is required to develop effective sex control techniques. Red spotted grouper were reared in indoor tanks and sampled every 5 days from 40 days post-hatch (DPH) to 130 DPH. Changes of gonadal tissues were examined and analyzed by means of histology. A pair of gonadal primordium has already existed underneath the kidney in the posterior part of the body cavity at 38 DPH when this study began. Gonadal primordia of 38, 40 DPH consisted of germ cells surrounded by a few somatic cells. The blood vessel was observed in the gonadal primordium at 45 DPH. The number of somatic cells and size of gonadal primordium increased age-dependently up to 60 DPH. Formation of ovarian cavity was obvious by two protuberant aggregations of somatic cells at 65 DPH. Completed ovarian cavity and oogonia were first observed in the gonad of one fish sample at 105 DPH. Based on these histological observations, it can be suggested that induction of primary male differentiation could be more successfully applied at around 60 DPH in this species.

Red spotted grouper (Epinephelus akaara) is one of the most popular and important grouper species for aquaculture in South-East and East Asia thanks to its fast growth and high market value. This species is known as a protogynous hermaphrodite which first differentiates into females and changes to males later. Natural sex change in this species occurs at 2 or 3 years of age. Many studies have been conducted, so far, to develop standardized methods for artificial sex reversal by treatment with sex steroid hormones (Bhandari et al., 2005; Lee et al., 2014). However, sex-changed male groupers showed testes with ovarian cavity, and their sperm production was very low. Induction of primary sex differentiation directly into males would be an alternative approach. Identification of the exact primary sex differentiation period is the prerequisite for this approach. Red spotted grouper were reared in indoor tanks and acclimatized at 25±2℃ under natural photoperiod. Fish were sampled at 60 days after hatching. The samples were processed for histological analysis using standard techniques in an automatic tissue processor, sectioned at 8 μm in thickness and stained with haematoxylin and eosin. The result of this study shows the morphological characteristics of gonadal primordium, and suggests that the timing of gonadal differentiation in red spotted grouper is at least 10 days earlier than the previous study of Lee et al. (2014) which observed gonadal primordium at 70 days after hatching.