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Dynamic changes of yak (Bos grunniens) gut microbiota during growth revealed by polymerase chain reaction-denaturing gradient gel electrophoresis and metagenomics

  • Nie, Yuanyang;Zhou, Zhiwei;Guan, Jiuqiang;Xia, Baixue;Luo, Xiaolin;Yang, Yang;Fu, Yu;Sun, Qun
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.7
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    • pp.957-966
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    • 2017
  • Objective: To understand the dynamic structure, function, and influence on nutrient metabolism in hosts, it was crucial to assess the genetic potential of gut microbial community in yaks of different ages. Methods: The denaturing gradient gel electrophoresis (DGGE) profiles and Illumina-based metagenomic sequencing on colon contents of 15 semi-domestic yaks were investigated. Unweighted pairwise grouping method with mathematical averages (UPGMA) clustering and principal component analysis (PCA) were used to analyze the DGGE fingerprint. The Illumina sequences were assembled, predicted to genes and functionally annotated, and then classified by querying protein sequences of the genes against the Kyoto encyclopedia of genes and genomes (KEGG) database. Results: Metagenomic sequencing showed that more than 85% of ribosomal RNA (rRNA) gene sequences belonged to the phylum Firmicutes and Bacteroidetes, indicating that the family Ruminococcaceae (46.5%), Rikenellaceae (11.3%), Lachnospiraceae (10.0%), and Bacteroidaceae (6.3%) were dominant gut microbes. Over 50% of non-rRNA gene sequences represented the metabolic pathways of amino acids (14.4%), proteins (12.3%), sugars (11.9%), nucleotides (6.8%), lipids (1.7%), xenobiotics (1.4%), coenzymes, and vitamins (3.6%). Gene functional classification showed that most of enzyme-coding genes were related to cellulose digestion and amino acids metabolic pathways. Conclusion: Yaks' age had a substantial effect on gut microbial composition. Comparative metagenomics of gut microbiota in 0.5-, 1.5-, and 2.5-year-old yaks revealed that the abundance of the class Clostridia, Bacteroidia, and Lentisphaeria, as well as the phylum Firmicutes, Bacteroidetes, Lentisphaerae, Tenericutes, and Cyanobacteria, varied more greatly during yaks' growth, especially in young animals (0.5 and 1.5 years old). Gut microbes, including Bacteroides, Clostridium, and Lentisphaeria, make a contribution to the energy metabolism and synthesis of amino acid, which are essential to the normal growth of yaks.

The Relation of Alexithymia, Somatic Complaints, Emotion and Vocabulary (감정표현불능증(Alexithymia), 신체적 호소, 정서 및 어휘의 관계)

  • Jeon, Hyun-Tae;Lee, Kuy-Haeng;Kim, Jae-Hyun;Kim, Han-Joo;Yoo, Yong-Jin;So, Kwang
    • Korean Journal of Psychosomatic Medicine
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    • v.8 no.1
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    • pp.58-64
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    • 2000
  • Objectives : This study aimed to examine a correlation between the somatic complaints, emotion, vocabulary and alexithymia as a component of personality in normal persons. Methods : 204 subjects were collected by age-based systematic sampling from the 662 persons without confirmed medical illness. We used the Korean version of 20-item Toronto Alexithymia Scale(TAS-20K) to measure alexithymia. The somatic complaints were checked by the list of somatic symptoms on the diagnostic criteria of somatization disorder and major depressive episode in DSM-IV. The vocabulary was evaluated by the total number of associating-words from the spontaneous association of word and the secondary association to given words. The anxiety and depression were evaluated using 5-point self-report scale. Results : 1) The degree of alexithymia was significantly correlated with the somatic complaints, anxiety, depression. 2) The somatic complaints were significantly correlated with the anxiety and depression. 3) The number of associating-words showed negative correlation with the age. 4) The degree of alexithymia was not correlated with the number of associating-words. Conclusion : The more degree of alexithymia increased, the more somatic complaints appeared. There was a significant correlation between the degree of alexithymia, anxiety and depression. But the degree of alexithymia was not correlated with the amount of vocabulary.

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IgG Humoral Immune Response to Extract Proteins of Malassezia Pachydermatis Isolated from a Dog with Atopic Dermatitis (Ad) (아토피 견에서 분리한 M. pachydermatis 추출 단백질에 대한 IgG 체액성 면역 반응의 연구)

  • Kim, Eun-Tae;Kim, Ha-Jung;Lim, Chae-Young;Park, Hee-Myung
    • Journal of Veterinary Clinics
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    • v.25 no.5
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    • pp.340-345
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    • 2008
  • Malassezia pachydermatis (M. pachydermatis) is a component of the normal cutaneous flora of the dog and atopic dermatitis (AD) is one of the most common diseases associated with Malassezia overgrowth in dogs. The purpose of this study was to investigate the humoral response (IgG) to extracts of M. pachydermatis of in a dog with AD. We used Western immunoblotting to identify allergens of M. pachydermatis. Gel electrophoresis of extracts proteins and immunoblotting of sera samples in both an atopic dog and a non-atopic dog were compared. Proteins of 18, 21, 26, 32, 34, 38, 40, 42, 46, 58, 64, 75, 85, and 120 kDa were observed in a serum of atopic dog. However, when serum of a non-atopic dog was used, protein bands were not identified except binding in 50 kDa protein. The results of this study indicate that atopic dogs with M. pachydermatis dermatitis may induce IgG response and also suggest that humoral response to M. pachydermatis could be important in pathogenesis of AD in dogs. However, further studies are required to identify roles of humoral response to M. pachydermatis in canine AD.

Neuropsychological Approaches to Mathematical Learning Disabilities and Research on the Development of Diagnostic Test (신경심리학적 이론에 근거한 수학학습장애의 유형분류 및 심층진단검사의 개발을 위한 기초연구)

  • Kim, Yon-Mi
    • Education of Primary School Mathematics
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    • v.14 no.3
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    • pp.237-259
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    • 2011
  • Mathematics learning disabilities is a specific learning disorder affecting the normal acquisition of arithmetic and spatial skills. Reported prevalence rates range from 5 to 10 percent and show high rates of comorbid disabilities, such as dyslexia and ADHD. In this study, the characteristics and the causes of this disorder has been examined. The core cause of mathematics learning disabilities is not clear yet: it can come from general cognitive problems, or disorder of innate intuitive number module could be the cause. Recently, researchers try to subdivide mathematics learning disabilities as (1) semantic/memory type, (2) procedural/skill type, (3) visuospatial type, and (4) reasoning type. Each subtype is related to specific brain areas subserving mathematical cognition. Based on these findings, the author has performed a basic research to develop grade specific diagnostic tests: number processing test and math word problems for lower grades and comprehensive math knowledge tests for the upper grades. The results should help teachers to find out prior knowledge, specific weaknesses of students, and plan personalized intervention program. The author suggest diagnostic tests are organized into 6 components. They are number sense, conceptual knowledge, arithmetic facts retrieval, procedural skills, mathematical reasoning/word problem solving, and visuospatial perception tests. This grouping will also help the examiner to figure out the processing time for each component.

Characterization of TNNC1 as a Novel Tumor Suppressor of Lung Adenocarcinoma

  • Kim, Suyeon;Kim, Jaewon;Jung, Yeonjoo;Jun, Yukyung;Jung, Yeonhwa;Lee, Hee-Young;Keum, Juhee;Park, Byung Jo;Lee, Jinseon;Kim, Jhingook;Lee, Sanghyuk;Kim, Jaesang
    • Molecules and Cells
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    • v.43 no.7
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    • pp.619-631
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    • 2020
  • In this study, we describe a novel function of TNNC1 (Troponin C1, Slow Skeletal and Cardiac Type), a component of actin-bound troponin, as a tumor suppressor of lung adenocarcinoma (LUAD). First, the expression of TNNC1 was strongly down-regulated in cancer tissues compared to matched normal lung tissues, and down-regulation of TNNC1 was shown to be strongly correlated with increased mortality among LUAD patients. Interestingly, TNNC1 expression was enhanced by suppression of KRAS, and ectopic expression of TNNC1 in turn inhibited KRASG12D-mediated anchorage independent growth of NIH3T3 cells. Consistently, activation of KRAS pathway in LUAD patients was shown to be strongly correlated with down-regulation of TNNC1. In addition, ectopic expression of TNNC1 inhibited colony formation of multiple LUAD cell lines and induced DNA damage, cell cycle arrest and ultimately apoptosis. We further examined potential correlations between expression levels of TNNC1 and various clinical parameters and found that low-level expression is significantly associated with invasiveness of the tumor. Indeed, RNA interference-mediated down-regulation of TNNC1 led to significant enhancement of invasiveness in vitro. Collectively, our data indicate that TNNC1 has a novel function as a tumor suppressor and is targeted for down-regulation by KRAS pathway during the carcinogenesis of LUAD.

Antimicrobial and Antitumor Activities of Seed Extracts of Camellia sinensis L. (차나무 종자 추출물의 항균활성 및 항종양활성)

  • Yoon, Won-Ho;Choi, Jae-Hoon;Lee, Keyong-Ho;Kim, Chang-Han
    • Korean Journal of Food Science and Technology
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    • v.37 no.1
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    • pp.108-112
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    • 2005
  • Antimicrobial and antitumor activities of Camellia sinensis L seed extracts were investigated. Seed extracts showed antifungal activities against Candida albicans IFO 1594 and Cryptococcus neoformans. Inhibition zone of 20 mm was shown by 70% ethanol extract against C. albicans IFO 1594 at 100 mg/mL. Antifungal activity of seed extract was not decreased by heating at 80 and $100^{\circ}C$ for 30 min or at $121^{\circ}C$ for 15 min, indicating heat-stability of seed component. Growth-inhibitory effects were observed in 70 and 10% of tumor cell line SK-OV-3 and normal ceil line NIH/3T3 at $50{\mu}g/mL$, respectively.

Determination of Tocopherol and Tocotrienol Contents in Rice Cooked with Various Cereals (밥의 종류에 따른 토코페롤 및 토코트리에놀 함량 분석 -연구노트-)

  • Kim, Yangsoo;Park, Soonriang;Lee, Young-Sang;Jung, Hwan;Koh, Kwangoh;Kim, Hee-Seon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.8
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    • pp.1289-1292
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    • 2005
  • The objective of this study was to characterize and determine contents of vitamin E isomers (alpha-, beta-, gamma-, delta-tocopherols and tocotrienols) in different steamed rice dishes, with or without other grains. Five different rice dishes were evaluated for the vitamin E nutritional value as major Korean staple foods. They were plain steamed rice (SR) and steamed rice mixed with barley (SRBa), red bean (SRRB), black bean (SRBB), or multi-grains (SRMG) containing a mixture of black rice, barley, red beans, and black beans. Vitamin E isomers were extracted from five grams of freeze-dried samples with hexane after saponification. An analytical method, using a normal-phase HPLC with a UV detector, was developed and used to determine the amount of each vitamin E component. The results showed that SR contained three vitamin E isomers (alpha-tocopherol, alpha-tocotrienol, and gamma-tocotrienol). Alpha-tocopherol and gamma-tocotrienol were de-tected from all samples while only SRBB contained beta-tocopherol and beta-tocotrienol. SRMG showed the highest (3.9$\mu$g/g dry wt) and SRRB showed the lowest alpha-tocopherol (1.3$\mu$g/g dry wt) contents. SRBB contained about 5 to 16 times more gamma-tocopherol (19.7$\mu$g/g dry wt) than othe.5. These results suggested that adding black bean or multi-grains can dramatically improve the vitamin E nutritional values compared to the plain steamed rice (SR). Information obtained from this study can be directly related to the amount of vitamin E intake and can be used to balance the diet for Koreans.

Influence of Polarization Behaviors on the ECM Characteristics of SnPb Solder Alloys in PCB (PCB에서의 ECM 특성에 미치는 SnPb 솔더 합금의 분극거동의 영향)

  • Lee Shin-Bok;Yoo Young-Ran;Jung Ja-Young;Park Young-Bae;Kim Young-Sik;Joo Young-Chang
    • Journal of the Microelectronics and Packaging Society
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    • v.12 no.2 s.35
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    • pp.167-174
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    • 2005
  • Smaller size and higher integration of electronic components make smaller gap between metal conducting layers in electronic package. Under harsh environmental conditions (high temperature/humidity), electronic component respond to applied voltages by electrochemically ionization of metal and metal filament formation, which lead to short failure and this phenomenon is termed electrochemical migration(ECM). In this work, printed circuit board(PCB) is used for determination of ECM characteristics. Copper leads of PCB are soldered by eutectic solder alloys. Insulation breakdown time is measured at $85^{\circ}C,\;85{\%}RH$. CAF is the main mechanism of ECM at PCB. Pb is more susceptible to CAF rather than Sn, which corresponds well to the corrosion resistance of solder materials in aqueous environment. Polarization tests in chloride or chloride-free solutions fur pure metal and eutectic solder alloys are performed to understand ECM characteristics. Lifetime results show well defined log-normal distribution which resulted in biased voltage factor(n=2) by voltage scaling. Details on migration mechanism and lifetime statistics will be presented and discussed.

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Development of a Korean Sexual Satisfaction Scale (한국 성인의 성만족 측정도구 개발)

  • Lee Byung-Sook;Koh Hyo-Jung;Kim Myung-Ae;Im Shin-Hwa
    • Journal of Korean Academy of Fundamentals of Nursing
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    • v.9 no.2
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    • pp.190-200
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    • 2002
  • Purpose; The purpose of this study was to develop an instrument to measure sexual satisfaction in Korean adults. Method: Development of items for the instrument was done by a collecting, reorganizing, reducing and selecting process based on the literature. A total of 19 items were selected in this process. For evaluation of the appropriateness of the 19 items, questionnaires were given to an expert group consisting of 18 professionals (7 men and 11 women). Eighteen items having a response of 70% in 'very good' item or 'good' item were included as items for the instrument. With these 18 items, a 5 point Likert scale was developed, and reliability and validity tests of the scale were done. The subjects for the test were 1,127 Korean adults living in 11 areas in Korea, Data were collected from Dec. 1, 2000 to Feb. 28, 2001 and analyzed with SPSS/PC+ 10.0 program. Item analysis and factor analysis were used to assess the reliability and validity of the scale. Chronbach alpha and corrected item-total correlation were obtained to verify internal consistency, and principal component analysis with the varimax rotation was used to identify structure validity of the developed tool. Result: Reliability of the scale was .9503 and corrected item-total correlations of the items were distributed from .5208 to 7899. As a result of the factor analysis, the items were categorized into three factors. The cumulative variance of the three factors was 67.1%, and all of the variances of the factors were above 5%. Because factor 1 (8 items) indicated a special feeling of psychological satisfaction with sexual activity, it was named 'psychological factor'. Factor 2 (8 items) indicated a feeling of satisfaction in the interaction with sexual partner through communication and variation, therefore it was named 'interactional factor'. Factor 3 (2 items) was about the frequency and duration of sexual intercourse and indicated physical dimensions of satisfaction, so it was named 'physical factor'. The average score for sexual satisfaction of the 1,127 subjects was 65.32 (${\pm}12.71$), and histogram of the score showed a normal distribution with skewness - .425. Conclusion: Finally, a Korean Sexual Satisfaction Scale(KSSS) was accepted as a 5 point Likert scale with 18 items after the statistical test for reliability and validity. In conclusion, KSSS is a useful instrument which is valid and reliable can be used to understand the sexuality of Korean adults.

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Effects of Nicotine on the Expression of Cell Cycle Regulatory Proteins of Human Gingival Fibroblasts (니코틴이 치은섬유아세포의 세포주기 조절 단백질 발현에 미치는 영향)

  • Kim, Tak;Kim, Jae-ho;Pi, Sung-Hee;Kim, Eun-Cheol;You, Yong-Ouk;You, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.31 no.3
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    • pp.597-610
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    • 2001
  • Normal gingival fibroblasts functioning is fundamental for the maintenance of periodontal connective tissue as well as wound healing. Nicotine have been found to affect DNA synthesis and cell proliferation, which appear to depend on the type of cells. This in vitro study was done to determine the effects of nicotine, a major component of tobacco, on cell proliferation, viability, activity, cell cycle distribution, and expression of cell cycle regulatory proteins in human gingival fibroblasts. Nicotine has been tested for 2 days or 4 days in 5 different concentrations; $0.1{\mu}g/ml$; $1{\mu}g/ml$; $10{\mu}g/ml$; $100{\mu}g/ml$; $1000{\mu}g/ml$. To assess cell proliferation and viability, viable and non-viable cells were counted by hemocytometer; to evaluate cellular activity, MTT assay was employed; to analyze cell cycle distribution, fluorescent propidium iodide-DNA complex were measured using fluorocytometer; to determine the expression of cell cycle regulatory proteins, western blot analysis was performed. After 2 days and 4 days incubation respectively, at concentrations of $1{\mu}g/ml$ - $1000{\mu}g/ml$, nicotine significantly inhibited proliferation comparing to non-supplemented controls. The cell viability was significantly decreased after 2 days and 4 days at concentrations of $1{\mu}g/ml$ - $1000{\mu}g/ml$ and at $10{\mu}g/ml$ - $1000{\mu}g/ml$ respectively. After 2 days and 4 days, the cellular activity was significantly decreased at concentrations of $10{\mu}g/ml$ - $1000{\mu}g/ml$. Treatment with $100{\mu}g/ml$ nicotine for 48 hours caused an increase in the proportion of G1-phase cells (from 46.41% to 53.46%) and a decrease in the proportion of S-phase cells (from 17.80% to 14.27%). The levels of cyclin $D_1$ and CDK 4 proteins in nicotine-treated fibroblasts were lower than that of controls, whereas the levels of p16 and pRB were higher than that of controls. These results suggest that the decrease of cell proliferation and lengthened Gap phases (G1) by nicotine may due to the increased expression of p16 and pRB as well as decreased expression of cyclin $D_1$ and CDK 4 in human gingival fibroblasts.

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