• Title/Summary/Keyword: GAD

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Characterization of γ-Aminobutyric acid(GABA) produced by a lactic acid bacterium from button mushroom bed

  • Lee, Yun-Seok;Song, Tae-Young;Kong, Won-Sik;Yoon, Min-Ho
    • Journal of Mushroom
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    • v.11 no.4
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    • pp.181-186
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    • 2013
  • ${\gamma}$-Aminobutyric acid(GABA) is a four carbon non-protein amino acid that has several well-known physiological functions, such as a postsynaptic inhibitory neurotransmitter in the brain and induction of hypotensive and tranquilizer effects. A lactic acid bacterium was isolated from button mushroom bed, which is showing high GABA productivity by TLC or HPLC analysis. The strain was identified as Lactobacillus hilgardii by analysis of 16S rDNA gene sequence. When the maximum production of GABA by L. hilgardii was investigated with various concentration of monosodium glutamate, the yield of GABA reached to be 53.65 mM at 1% mono sodium glutamate (MSG) in flask cultivation. A Glutamate decarboxylase (GAD) enzyme, which was known to convert MSG to GABA, was purified from a cell-free extract of L. hilgardii and the molecular weights of purified GAD was estimated to 60,000 by SDS-PAGE. The optimum pH and temperature of GAD were at pH4.6 and at $37^{\circ}C$, respectively. The GAD activity was increased by the addition of sulfate ions such as ammonium sulfate, sodium sulfate and magnesium sulfate, indicating that the increase of hydrophobic interaction causes the increase of GAD activity.

Stress Coping Strategies and Cognitive Characteristics of Somatic Symptom Perception in Patients with Generalized Anxiety Disorder (범불안장애 환자의 스트레스 대처방식과 신체증상 지각에 대한 인지적 특성)

  • Kim, Yong;Park, Jong-Il;Park, Tae-Won;Chung, SangKeun;Yang, Jong-Chul
    • Anxiety and mood
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    • v.13 no.2
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    • pp.100-107
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    • 2017
  • Objective : The purpose of this study was to investigate strategies used to cope with stress and the cognitive characteristics of somatic symptom perception in patients with generalized anxiety disorder(GAD). Methods : A total of 55 patients meeting DSM-5 criteria for GAD and 55 normal controls were recruited for participation in this study. We evaluated subjects using The Way of Stress Coping Questionnaire (SCQ), Somato-Sensory Amplification Scale (SSAS), Symptom Interpretation Questionnaire (SIQ), and the Generalized Anxiety Disorder for 7 Item (GAD-7). We analyzed data using an independent t-test and Pearson's correlation analysis. Results : In terms of SCQ, GAD patients presented significantly lower scores on seeking social support and higher scores on wishful thinking than normal controls. GAD patients had significantly greater amplification of physical sensation in SSAS and higher scores in physical, psychological interpretation subsets of SIQ than normal controls. GAD-7 scores were positively correlated with physical interpretation scores on SIQ. Conclusion : Results reveal that patients with GAD have insufficient coping strategies for stress, greater amplification of body sensations, and tendency towards a physical, psychological interpretation of somatic symptoms.

Characterization of a Glutamate Decarboxylase (GAD) from Enterococcus avium M5 Isolated from Jeotgal, a Korean Fermented Seafood

  • Lee, Kang Wook;Shim, Jae Min;Yao, Zhuang;Kim, Jeong A;Kim, Hyun-Jin;Kim, Jeong Hwan
    • Journal of Microbiology and Biotechnology
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    • v.27 no.7
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    • pp.1216-1222
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    • 2017
  • To develop starters for the production of functional foods or materials, lactic acid bacteria producing ${\gamma}-aminobutyric$ acid (GABA) were screened from jeotgals, Korean fermented seafoods. One isolate producing a high amount of GABA from monosodium $\text\tiny{L}$-glutamate (MSG) was identified as Enterococcus avium by 16S rRNA gene sequencing. E. avium M5 produced $18.47{\pm}1.26mg/ml$ GABA when incubated for 48 h at $37^{\circ}C$ in MRS broth with MSG (3% (w/v)). A gadB gene encoding glutamate decarboxylase (GAD) was cloned and overexpressed in E. coli BL21 (DE3) using the pET26b (+) expression vector. Recombinant GAD was purified through a Ni-NTA column and the size was estimated to be 53 kDa by SDS-PAGE. Maximum GAD activity was observed at pH 4.5 and $55^{\circ}C$and the activity was dependent on pyridoxal 5'-phosphate. The $K_m$ and $V_{max}$ values of GAD were $3.26{\pm}0.21mM$ and $0.0120{\pm}0.0001mM/min$, respectively, when MSG was used as a substrate. Enterococcus avium M5 secretes a lot of GABA when grown on MRS with MSG, and the strain is useful for the production of fermented foods containing a high amount of GABA.

Properties of a Novel Glutamate Decarboxylase (GAD) from Levilactobacillus brevis B737 Isolated from Cabbage Kimchi

  • Tae Jin Kim;Min Jae Kim;Bong Sin Kim;Ji Yeon Yoo;Yun Ji Kang;Jeong Hwan Kim
    • Microbiology and Biotechnology Letters
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    • v.50 no.3
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    • pp.319-327
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    • 2022
  • γ-Aminobutyric acid (GABA) is a multi-functional compound with broad applications for food industry. GABA producing bacteria were isolated from cabbage kimchi. Among them, B737 was the best GABA producer when culture supernatants were analyzed by TLC. B737 was identified as Levilactobacillus brevis by 16S rRNA gene sequencing. Its glutamate decarboxylase (GAD) gene was cloned by PCR and the nucleotide sequence determined. B737 GAD consisting of 485 amino acids is the largest in size among GADs reported from LAB so far. gadB from L. brevis B737 was overexpressed in Escherichia. coli BL21(DE3) using pET26b(+).pET26b(+). The recombinant GAD was purified and its size was 55 kDa by SDS-PAGE. Maximum GAD activity was observed at pH 5 and 40℃ and the activity was dependent on pyridoxal 5'-phosphate. Km and Vmax of recombinant GAD were 6.2 ± 0.06 mM and 0.34 ± 0.002 mM/min, respectively. L. brevis B737 can be used as a starter for fermented foods with high GABA contents.

Evaluation of reference value of anti-Glutamic acid decarboxylase antibody for cerebrospinal fluid (뇌척수액에서 항 Glutamic acid decarboxylase 항체검사의 참고치 설정)

  • Park, Min-Ho;Shin, Sun-Young;Youn, Tae-Seok;Shin, Hi-Jung;Noh, Gyeong-Woon
    • The Korean Journal of Nuclear Medicine Technology
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    • v.21 no.2
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    • pp.28-30
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    • 2017
  • Purpose Anti-Glutamic acid decarboxylase antibody test (GAD Ab) has been used as a predictor of type 1 diabetes. GAD Ab has also been shown to be highly potent in cerebrospinal fluid (CSF) of patients with suspected diabetic peripheral neuropathy. Recently, it has been known that clinical significance of GAD Ab using CSF is useful for the neurological disorders. However, the reference value of anti-GAD Ab has been provided only for serum. In this experiment, we estimated the reference value of anti-GAD antibody for CSF in neurological patients. Materials and Methods A total of 211 neurological patients were enrolled. Serum and CSF were analyzed by radioimmunoassay (RIA) using commercial RIA anti-GAD Ab kit (RSR, London, United Kingdom). Normal saline was used as the normal CSF control because CSF is most similar to 0.9% normal saline. Results The mean value of normal CSF control was 1.97 U/mL, and two standard deviations (2SD) was 1.44 U/mL. Based on this data, the expected reference range of CSF could be estimated from 0.54 U/mL to 3.40 U/mL Conclusion The reference range of normal CSF control using normal saline obtained with Hoffmann's method. However, there will be a need to validate the CSF reference values using human normal CSF.

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The Effect of the Transmission of Coronavirus Disease-2019 on the Mentality of Parents and Children After the First Wave of Infections (1차 확산기 이후 코로나바이러스감염증-2019의 전파가 부모와 아동의 심리에 미치는 영향)

  • Kim, Jeongyeon;Lee, Koeun;Nam, Okhyung;Lee, Hyo-seol;Choi, Sungchul;Kim, Kwangchul;Kim, Misun
    • Journal of the korean academy of Pediatric Dentistry
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    • v.48 no.3
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    • pp.269-279
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    • 2021
  • The purpose of this study is to evaluate the effect of the spread of a new type of coronavirus infection (COVID-19) on the mental state in school-age children and parents focusing on the aspects of sleep disorders and depression. A questionnaire survey was conducted for 123 parents and 108 school-age children who visited Department of Pediatric Dentistry, Kyung Hee University Dental Hospital at Gangdong from April 2, 2020 through April 25, 2020, via the direct writing method. Participants were assessed with Pittsburgh Sleep Quality Index, Generalized Anxiety Disorder (GAD)-7, Center for Epidemiology Scale for Depression. Logistic regressions were used with a level of significance of 5%. The prevalence of GAD, depression, and poor sleep in parents were 34.1%, 17.1% and 44.7%, respectively. The prevalence of GAD in children was 20.4%. Logistic regression showed that stress from Emergency Alert Messages about COVID-19 was associated with GAD and depression in parents. In children, the degree of emotional change after COVID-19 was associated with GAD. This study confirmed that there was a change in the psychological status of children and guardians due to the epidemic of coronavirus disease-2019, and it would be necessary to consider their psychological status during dental treatment.

Analysis of Dietary Behavior and Related Factors According to the Level of Generalized Anxiety Disorder in Adolescents: Using the 16th-18th(2020-2022) Korea Youth Risk Behavior Survey (청소년의 범불안장애 수준에 따른 식생활 행태 및 관련 요인 분석: 제16차-제18차 (2020년-2022년) 청소년건강행태조사를 이용하여)

  • Kye, Eun Seul;Kim, Yookyung;Lee, Jung Woo
    • Journal of Korean Home Economics Education Association
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    • v.35 no.4
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    • pp.33-45
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    • 2023
  • The purpose of this study is to investigate the association between dietary behavior and related factors according to the generalized anxiety disorder (GAD) in adolescents. The results of analyzing demographic characteristics according to the GAD showed that gender, age, BMI, type of school, residence type, father's nationality, smoking and drinking experience, perceived health status, perception of body shape, economic status, academic achievement, physical activity, sedentary time and smartphone usage time were related to the GAD (p<0.01). The results of analyzing dietary behavior according to the GAD showed that the frequency of water intake, sweetened beverage intake, fast food intake, fruit intake and breakfast intake were related to the GAD (p<0.001). The results of analyzing depression and suicidality according to the GAD showed that depression, suicidal ideation, planning, and attempts were all related to the GAD (p<0.001). Even after adjusting for factors corresponding to demographic characteristics, it was confirmed that the level of GAD had a significant effect on depression and suicidality. This study is significant in that it provided basic data on adolescent mental health problems by analyzing dietary behavior and related factors according to the level of GAD.

Production of γ-Aminobutyric Acid Using Immobilized Glutamate Decarboxylase from Lactobacillus plantarum (Lactobacillus plantarum 유래 글루탐산 탈탄산효소의 고정화를 이용한 γ-aminobutyric acid의 생산)

  • Lee, Sang-Jae;Lee, Han-Seung;Lee, Dong-Woo
    • Microbiology and Biotechnology Letters
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    • v.43 no.3
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    • pp.300-305
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    • 2015
  • The glutamate decarboxylase gene (gadB) from Lactobacillus plantarum WCFS1 was cloned and expressed as an N-terminal hexa-histidine-tagged fusion protein in Escherichia coli BL21 (DE3) as the host strain. Purified glutamate decarboxylase (GAD) was immobilized onto porous silica beads by covalent coupling. The pH dependence of activity and stability of the immobilized GAD was significantly altered, when compared to those of the free enzyme. Immobilized GAD was stable in the range of pH 3.5 to 6.0. The resulting packed-bed reactor produced 41.7 g of γ-aminobutyric acid/l·h at 45℃.

Effects and Optimization of Gamma-Amino Butyric Acid (GABA) Production Process using Glutamate Decarboxylase (GAD) (Glutamate Decarboxylase (GAD)를 이용한 Gamma-Amino Butyric Acid (GABA) 생산 및 최적화)

  • Kim, Eui Jin;Lee, Jung-Heon
    • KSBB Journal
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    • v.29 no.6
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    • pp.426-431
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    • 2014
  • pH controlled batch reactor and bubble column reactors have been developed in this research. They were used to produce high concentration of GABA and to determine optimal pH for GABA production. Glutamate decarboxylase (GAD) was isolated from recombinant E. coli and used for GABA production from monosodium glutamate (MSG). pH control was inevitable because the pH increased with MSG consumption. GAD showed highest activity at acidic conditions at pH 5.5 but the optimal pH for GABA production was pH 6.0. When 1.5 mole of MSG was used as reactant, the 1.05 mole of GABA was produced after 10 hrs batch reaction. Using bubble column reactors, 80 % of MSG was converted to GABA for 6 hrs reaction and 1.2 mole of GABA was produced.

Contents and Severity of Worries in Outpatients with Generalized Anxiety Disorder Compared to Normal Control (범불안장애 환자와 일반인의 걱정 내용 및 심각도의 비교)

  • Kim, Hwigon;Kim, Daeho;Oh, Daeyoung;Seo, Ho Jun;Huh, Hyu Jung;Chae, Jeong Ho
    • Anxiety and mood
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    • v.11 no.1
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    • pp.47-53
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    • 2015
  • Objective : Generalized anxiety disorder (GAD) is a common anxiety disorder characterized by excessive worries and anxiety symptoms. To distinguish between pathological and normal worries, this study compared the contents and severity of worries among patients with GAD with those of matched non-psychiatric control Methods : Thirty outpatients with GAD receiving treatment at the psychiatric department of Hanyang University Guri Hospital were recruited. As a matched control group, we selected 30 adults without psychiatric history who participated in the Anxiety Disorder Survey, during the Mental Health Exposition held in Seoul in April, 2014. Matched demographic data were age, sex, education, and household income. Self-questionnaires included demographic data, contents of worry (10 categories of worries and each severity), and the Pennsylvania State Worry Questionnaire (PSWQ). Results : There was no statistical difference in the contents of worry between patient and control groups. However, the total PSWQ scores significantly higher in the GAD group. Conclusion : Our findings suggest that patients and non-patients worry about the similar contents and worries in GAD cannot be differentiated by what people worry about. Only the severity of worry was a distinguishable feature. Our finding supported inclusion of the diagnostic criterion of "excesslve wornes" in GAD.