• Title/Summary/Keyword: Gene detection

Search Result 1,202, Processing Time 0.026 seconds

Neuroprotective effects of hesperetin on H2O2-induced damage in neuroblastoma SH-SY5Y cells

  • Ha-Rin Moon;Jung-Mi Yun
    • Nutrition Research and Practice
    • /
    • v.17 no.5
    • /
    • pp.899-916
    • /
    • 2023
  • BACKGROUND/OBJECTIVES: Oxidative stress is a fundamental neurodegenerative disease trigger that damages and decimates nerve cells. Neurodegenerative diseases are chronic central nervous system disorders that progress and result from neuronal degradation and loss. Recent studies have extensively focused on neurodegenerative disease treatment and prevention using dietary compounds. Heseperetin is an aglycone hesperidin form with various physiological activities, such as anti-inflammation, antioxidant, and antitumor. However, few studies have considered hesperetin's neuroprotective effects and mechanisms; thus, our study investigated this in hydrogen peroxide (H2O2)-treated SH-SY5Y cells. MATERIALS/METHODS: SH-SY5Y cells were treated with H2O2 (400 µM) in hesperetin absence or presence (10-40 µM) for 24 h. Three-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assays detected cell viability, and 4',6-diamidino-2-phenylindole staining allowed us to observe nuclear morphology changes such as chromatin condensation and apoptotic nuclei. Reactive oxygen species (ROS) detection assays measured intracellular ROS production; Griess reaction assays assessed nitric oxide (NO) production. Western blotting and quantitative polymerase chain reactions quantified corresponding mRNA and proteins. RESULTS: Subsequent experiments utilized various non-toxic hesperetin concentrations, establishing that hesperetin notably decreased intracellular ROS and NO production in H2O2-treated SH-SY5Y cells (P < 0.05). Furthermore, hesperetin inhibited H2O2-induced inflammation-related gene expression, including interluekin-6, tumor necrosis factor-α, and nuclear factor kappa B (NF-κB) p65 activation. In addition, hesperetin inhibited NF-κB translocation into H2O2-treated SH-SY5Y cell nuclei and suppressed mitogen-activated protein kinase protein expression, an essential apoptotic cell death regulator. Various apoptosis hallmarks, including shrinkage and nuclear condensation in H2O2-treated cells, were suppressed dose-dependently. Additionally, hesperetin treatment down-regulated Bax/Bcl-2 expression ratios and activated AMP-activated protein kinase-mammalian target of rapamycin autophagy pathways. CONCLUSION: These results substantiate that hesperetin activates autophagy and inhibits apoptosis and inflammation. Hesperetin is a potentially potent dietary agent that reduces neurodegenerative disease onset, progression, and prevention.

Case Report on NTBC Treatment of Type 1 Tyrosinemia Diagnosed through Newborn Screening (신생아 선별검사를 통해 진단된 1형 타이로신혈증의 NTBC 치료 사례 보고)

  • Ji Eun Jeong;Hwa Young Kim;Jung Min Ko
    • Journal of The Korean Society of Inherited Metabolic disease
    • /
    • v.23 no.2
    • /
    • pp.39-44
    • /
    • 2023
  • Hereditary tyrosinemia type 1 (HT-1) is a metabolic disorder caused by biallelic pathogenic variants in the fumarylacetoacetate hydrolase (FAH) gene, which impairs the function of the FAH enzyme, resulting in the accumulation of tyrosine's toxic metabolites in hepatocytes and renal tubular cells. As a consequence, individuals with HT-1 exhibit symptomatic manifestations. Rapid diagnosis and treatment of HT-1 can prevent short-term death and long-term complications. A 15-day-old boy presented to the outpatient department with elevated levels of tyrosine on his newborn screening tests conducted at the age of 3 and 10 days, respectively. Further blood tests revealed increased levels of alpha-fetoprotein and amino acids including tyrosine and threonine. Urine organic acid tests indicated a significant elevation in tyrosine metabolites, as well as the presence of succinylacetone (SA), which led to the diagnosis of HT-1. Two pathogenic and likely pathogenic variants of FAH compatible with HT-1 were also detected. He began a tyrosine-restricted diet at one month old and received nitisinone (NTBC) at two months old. With continued treatment, the patient's initially elevated AFP level, detection of SA in the urine, and mild hepatomegaly showed improvement. During four years and seven months of treatment, there were no exceptional complications apart from an increase in tyrosine levels and a delay in speech. We report a case of tyrosinemia type 1 detected through newborn screening, treated with dietary restriction and NTBC, with a good prognosis.

  • PDF

Detection of Copy Number Variation of the KIT Gene in the Landrace Breed using an Quantitative Oligonucleotide Ligation Assay(qOLA) (Quantitative Oligonucleotide Ligation Assay(qOLA)를 이용한 Landrace 품종의 KIT 유전자 반복수 변이 탐지)

  • Seo, B.Y.;Kim, J.H.;Nahm, D.W.;Yoo, C.K.;Lee, S.H.;Lee, J.B.;Lim, H.T.;Jung, E.J.;Cho, I.C.;Heo, K.N.;Jeon, J.T.
    • Journal of Animal Science and Technology
    • /
    • v.49 no.5
    • /
    • pp.559-568
    • /
    • 2007
  • Recently, copy number variations (CNV) of genes or genomic segments have been intensively studied and various analysis methods have been developed. In this study, quantitative oligonucleotide ligation assay (qOLA) was applied to investigate CNV of KIT gene in the Landrace breed. A combined assay using qOLA and pyrosequencing, 6 genotype classes, I1/I1 or I3/i (IBe), I1/I2 or I3/IP, I1/I3, I1/IP or I2/i (IBe), I2/I2and I2/IP, were identified from 44 Landrace pigs. Genotype assignment using grouping features of measurements on a scatter plot showed 100% agreement with those using a statistical assignment by PROC FASTCLUS procedure implemented in the SAS package. Two versions (3100 and 3130) of ABI sequencers gave the same genotyping results, indicating there was no influence on qOLA by different versions of instrument, however, the means of standard deviation and coefficient of variation from the qOLA on a ABI 3130 (2.33 and 4.10) was lower than those from the qOLA on a ABI 3100 (2.67 and 4.81). Effect of proteinase K treatment on the PCR product followed by qOLA was very clear because noise peaks were disappeared and the observed ration fit better to the reference ratio corresponding to each genotype.

Development of Detection Method for Oilfish (Ruvettus pretiosus and Lepidocybirium flavobrunneum) as a Food Materials not Usable in Foods (식품원료로 사용금지 대상인 기름치 (기름갈치꼬치 및 흑갈치꼬치) 판별법 개발)

  • Park, Yong-Chjun;Kim, Mi-Ra;Jung, Yong-Hyun;Shin, Joon-Ho;Kim, Kyu-Heon;Lee, Jae-Hwang;Cho, Tae-Yong;Lee, Hwa-Jung;Lee, Sang-Jae;Han, Sang-Bae
    • Journal of Food Hygiene and Safety
    • /
    • v.28 no.1
    • /
    • pp.50-55
    • /
    • 2013
  • Since 1 June 2012, it is prohibited to sell oilfish as a food material but there are still many illegal cases of selling oilfish as if it is tuna or grilled Patagonian toothfish. So it is absolutely crucial to construct the system to distinguish the real food material from oilfish. There are two sorts of oil fish called Ruvettus pretiosus and Lepidocybirium flavobrunneum involved in Percifomes order and Gempylidae class. 16S DNA gene region in mitochondria was selected to design the specific primers. For design species-specific primer, the theoretical experiment were performed for the sequences of R. pretiosus, L. flavobrunneum, Thunnus thynnus, Thunnus albacores, Makaira mitsukurii and Xiphias gladius, registered at the Gene bank from the National Centre for Biotechnology Information, using BioEdit 7.0.9.0. program. Through the analysis of the result from experiments, it was possible to design the 4 kinds of primers to distinguish R. pretiosus and L. flavobrunneum. As a comparison group, 3 kinds of tuna and 4 kinds of billfishes were selected and experimental verification was performed. As a result, for R. pretiosus and L. flavobrunneum, R.P-16S-006-F/R.P-16S-008-R and L.F-16S-004-F/L.F-16S-006-R primers were selected eventually and PCR condition was established. In addition, 178bp and 238bp of PCR products were confirmed from the established condition and non-specific band was not amplified among similar species. Therefore, the species-specific primers developed in this study would be very useful and used in various ways such as internet shopping mall and illegal distributions with fast and scientific results.

Genetic Properties and Antimicrobial Resistance of Campylobacter jejuni Isolates from Diarrhea Patients in Gyeonggi-do (경기도내에서 분리한 캠필로박터 제주니균의 유전적특성 및 항생제내성 연구)

  • Hur, Eun-Seon;Park, Po-Hyun;Kim, Jong-Hwa;Son, Jong-Sung;Yun, Hee-Jeong;Lee, Yea-Eun;Choi, Yun-Sook;Yoon, Mi-Hye;Lee, Jong-Bok
    • Korean Journal of Microbiology
    • /
    • v.49 no.3
    • /
    • pp.228-236
    • /
    • 2013
  • Campylobacter jejuni is one of important food-borne pathogens causing human gastroenteritis. We isolated 42 strains of C. jejuni from diarrhea patients and 4 food-poisoning outbreaks in 2010, Gyeonggi-do. In this study, 42 strains were tested for genetic characteristics, the serotype distribution and antimicrobial resistant rate. The presence of hipO (100%), cdtB (100%), and mutated gyrA (95.2%) genes was detected in C. jejuni by polymerase chain reaction (PCR). Detection of mutated gyrA gene correlated with ciprofloxacin resistance. Forty isolates had mutated gyrA gene and were actually resistant to ciprofloxacin. Furthermore, comparing the gyrA DNA sequence data, ciprofloxacin-resistant isolates had a mutation of the DNA sequence from ACA (threonine) to ATA (isoleucine). But 41 strains (97.6%) of patient isolates were susceptible to erythromycin and azithromycin. A total of 35.7% among 42 C. jejuni isolates were identified into 4 different serotypes. The serotype distribution of C. jejuni strains were shown to be HS2(B), HS3(C), HS4(D), HS19(O). To investigate the genotypes of C. jejuni isolated in Gyeonggi province, repetitive sequence polymerase chain reaction (rep-PCR) analysis and SmaI-digested pulsed-filed gel electrophoresis (PFGE) profile analysis were performed. From the PFGE analysis of 42 C. jejuni strains, 12 clusters of PFGE profile were obtained. On the other hand, 11 clusters of rep-PCR profile were obtained from 42 strains of C. jejuni.

Characteristics of Vibrio vulnificus Isolated in Incheon (인천지역에서 분리된 비브리오 패혈증균의 특성)

  • Oh, Bo-Young;Kim, Jung-Hee;Gong, Young-Woo;JeGal, Seung;Kim, Hye-Yeung;Lee, Mi-Yeon;Hwang, Kyoung-Wha;Koh, Yeon-Ja;Lee, Jae-Mann;Go, Jong-Myoung;Kim, Yong-Hee
    • Korean Journal of Microbiology
    • /
    • v.43 no.4
    • /
    • pp.256-263
    • /
    • 2007
  • We performed the biochemical characteristics, molecular epidemiologocal analysis, and drug susceptibility test on V. vulnificus isolated from environmental sources in Incheon. For this study, 233 strains were isolated from seawater, sediment, shellfish. V. vulnificus isolates were divided into 15 biochemical groups, which were positive for ONPG and Amygdalin test. Among the 209 strains, 206 (98.6%) strains and 110 (52.6%) strains revealed positive for vvhA and viuB gene, and the viuB gene detection rates of V. vulnificus from seawater, shellfish and sediment were 48%, 48.5% and 61.6%, respectively. From disc diffusion test on 175 isolates, most of strains were sensitive to Imipenem (100.0%), Sulfamethoxazole/trimethoprim (98.9%), Tetracycline, Ciprofloxacin (98.3%), Ampicillin/sulbactam (97.1%), Ohloramphenicol (96.6%), Cefepime (94.9%) and Ceftriaxone (94.8%), multi-drug resistance rates was 31.5% of seawater, 34.4% of sediment and 29.2% of shellfish. PFGE was performed on 233 V. vulnificus isolates with the objective of investigating the extent of genetic diversity of these isolates in our region. We could find that at least 126 different PFGE patterns were generated according by 90% of similarity and 13 clusters by 58% of similarity. The major cluster was type I (44.6%) during the most of the year, and type J was frequent pattern in June and October. There were 9 distinct PFGE types in July, 8 types in August, 7 types in June, 6 types in September, 5 types in October 3 types in May and 1 type in March.

Studies on the Physiological Chemistry of Flower Organ and Seed in Ginseng Plant. IV. Variation of Free Amino Acids in the Flower and Seeds of the $F_1$ Plants of the Combinations Panax ginseng ${\times}$ Panax quinquefolium and Panax ginseng ${\times}$ Panax japonicus. (인삼종자형성에 대한 생리화학적 연구 IV. 고려인삼과 미국인삼 및 고려인삼과 죽절인삼 $F_1$의 화기 및 종자 형성과정에 있어서의 유리아미노산의 소장)

  • Jong-Kyu Hwang;Hee-Chun Yang
    • KOREAN JOURNAL OF CROP SCIENCE
    • /
    • v.14
    • /
    • pp.165-172
    • /
    • 1973
  • The sterile phenomenon is frequently found in the inter-species hybrids of ginseng as in other plants. It is known that among the hybrids between Panax Ginseng (PG) and Panax Quinquefolium (PQ), and between Panax Ginseng and Paxax Japonicus (PI), PG${\times}$PI is fertile only very rarely, while PG ${\times}$ PQ is always sterile. Therefore, in order to clarify the relationship between this sterility phenomenon and the metabolism of free amino acids, the changes of free amino acids through the formation of the flower organs and seeds of two hybrids, PG ${\times}$ PQ and PG ${\times}$ PI were investigated by thin layer chromatography. The results are summarized as follows: 1. Distinct differences in the quantity and number of free amino acids were recognized between PG ${\times}$ PQ, PG ${\times}$ PI and their parent plants. From the hybrid PG ${\times}$ PQ, 19 kinds of ninhyrin sensitive substances were detected in all. They were (1) 17 amino acids: alanine, valine, leucine, phenylalanine, proline, hydroxy-proline, serine, threonine, tyrosine, aspartic acid, glutamic acid, lysine, arginine, ${\gamma}$-amino butyric acid, ${\beta}$-alanine, cysteic acid and tryptophan, and (2) two amides: asparagine and glutamine. From the hybrid PG ${\times}$ PI, in addition to the above 19 substances, methionine and one unknown substance were detected. 2. Generally, alanine, as partie acid, glutamic acid, cysteic acid and asparagine were detected in large amounts in the two hybrids as in PG, PG and PJ but it was a noticeable fact concerning these two hybrids that the largest quantity of asparagine was found at microspore satge and pollen mature stage. 3. The decrease of cysteic acid in the two hybrids at the red ripened stage was the same as in PQ and PJ but opposite to the change in PG. The detection of methionine in PG ${\times}$ PJ was worthy of notice. 4. The change of proline was conspicuously different from that in their parent plants. It was detected as a trace of color at the micros pore stage while asparagine was detected in the greatest amount at that time. It is well known that the quantity of proline is closely related to the sterility of plant. This fact was also found true in the formation of ginseng seeds. It was reported as well that asparagine accumulated when proline decreased. 5. The deficiency of proline seemed to be closely related with the sterility of hybrids and with the degradation of pollen in anther. 6. The difference in the changes of free amino acids between the selfed lines of PG, PQ and PJ, and their hybrids seemed to be caused by the transformation of gene-action system by hybridization. On these phenomena along with proline metabolim and its physiological role in seed formation further studies are required.

  • PDF

Molecular Epidemiological Characteristics of Vibrio Parahaemolyticus as Recently wilde-spreaded in Korea (최근 한국에서 유행하는 장염비브리오균의 분자 역학적 특성)

  • 김상숙;이희무;이중복
    • KSBB Journal
    • /
    • v.18 no.6
    • /
    • pp.522-528
    • /
    • 2003
  • The purpose of this study is to inquire into molecular epidemiological characteristics of Vibrio parahaemolyticus. For this study, 120 strains(120 strains of Vibrio parahaemolyticus sampled from diarrhea patients) were examined and analyzed for biochemical characteristics, TDH (thermostable direct hemolysin) antibiotics sensitivity and detection of toxR, gyrE, tdh, and tds gents. G-S PCR (Group Specific Polymerase), PFGE (Pulsed-field Gel Electrophoriesis) methods were performed on the materials from patients were results. 1 Vibrio parahaemolyticus didn't grow in 0% density of NaCl, but the fact was found that those grew in 8% density of NaCl. 2. O:K serotypes of Vibrio parahaemolyticus was turned up in domestic patients was 17 types. Among those O3:K6 was the most, it was 68.3%. 3. In 18 kinds of antibiotic tests resistant against Ampicillin, Ticacillin was comparatively high. the case of resistant against Ampicillin, Ticacillin, Vancomycin at the multiple resistant was 52.5%. 4. Toxin gene tdh had only 109 strains among 120 ones isolated from patients held the genes of 199bp size, and 11 strains was negativity 5. In the test of Kanagawa toxic productivity, 107 strains among strains isolated from patients appeared to be positivity reaction 6. The strain that held trh toxin was only 3, and those among test strains had the genes of 250bp size and that had tdh, trh genes at a time were 3 strains, and TDH toxic productivity of those were 16 times, and it was weak. 7. Group Specific-PCR appeared to be useful in the confirmation of O3:K6 serotype interrelations. 8. Three strains which showed difference of 7 DNA sequence even in the same serotype were detected by the result of analyzing the regular gene, toxRS DNA sequence. These strains are differ from general strains which carry infection easily. 9. These mutual dose epidemiological relations were classified into smaller-parts through PFGE method. As a result of such classify, 3 findings were found. V. parahaemolyticus sampled from diarrhea patients were classified into 3 types. And third, the result obtained through PFGE method can be used as a useful tool in a point of molecular-epidemiological view.

Effective Method for Extraction of Cell-Free DNA from Maternal Plasma for Non-Invasive First-Trimester Fetal Gender Determination: A Preliminary Study

  • Lim, Ji-Hyae;Park, So-Yeon;Kim, Shin-Young;Kim, Do-Jin;Kim, Mee-Jin;Yang, Jae-Hyug;Kim, Moon-Young;Kim, Min-Hyoung;Han, Ho-Won;Choi, Kyu-Hong;Ryu, Hyun-Mee
    • Journal of Genetic Medicine
    • /
    • v.7 no.1
    • /
    • pp.53-58
    • /
    • 2010
  • Purpose: To find the most effective method for extraction of cell-free DNA (cf-DNA) from maternal plasma, we compared a blood DNA extraction system (blood kit) and a viral DNA extraction system (viral kit) for non-invasive first-trimester fetal gender determination. Materials and Methods: A prospective cohort study was conducted with maternal plasma collected from 44 women in the first-trimester of pregnancy. The cf-DNA was extracted from maternal plasma using a blood kit and a viral kit. Quantitative fluorescent-polymerase chain reaction (QF-PCR) was used to detect the SRY gene and AMEL gene. The diagnostic accuracy of the QF-PCR results was determined based on comparison with the final delivery records. Results: A total of 44 women were tested, but the final delivery record was only obtained in 36 cases which included 16 male-bearing and 20 female-bearing pregnancies. For the blood kit and viral kit, the diagnostic accuracies for fetal gender determination were 63.9% (23/36) and 97.2% (35/36), respectively. Conclusion: In non-invasive first-trimester fetal gender determination by QF-PCR, using a viral kit for extraction of cf-DNA may result in a higher diagnostic accuracy.

Carriage Rates and Serogroups of Neisseria meningitides in Children Attending Day Care Centers (유아원에 다니는 소아의 구인강 내 수막구균 보균율과 혈청군)

  • Kim, Nam Hee;Lee, Jina;Lee, Jung Won;Lee, Soo Young;Choi, Eun Hwa;Kim, Kyung Hyo;Kim, Eui Chong;Kang, Jin Han;Lee, Hoan Jong
    • Pediatric Infection and Vaccine
    • /
    • v.16 no.1
    • /
    • pp.31-39
    • /
    • 2009
  • Purpose : Neisseria meningitides is one of the most common causative pathogens of bacteremia and meningitis. Recently protein-conjugated vaccines have been developed and included in the routine vaccination schedule in a few countries. In Korea, carriage rates of N. meningitides among healthy adults have been reported. However, systematic data for childhood carriage rates are not available. This study was performed to evaluate the carriage rates of N. meningitides and the serotype distribution among healthy children attending day care centers. Methods : During the period of January through May 2005, nasopharyngeal swabs and culture were obtained from 904 children attending 13 different day care centers located in Seoul and Gyeonggi Province. The Vitek NHI card was used to identify N. meningitides and the crgA gene was detected via polymerase chain reaction (PCR). Serotype determination was performed by agglutination test using N. meningitides antisera to serotypes A, B, C, D, 29E, W135, X, Y, and Z. PCR for detection of the org2 and saiD gene confirmed serotypes A, B, C, W135, and Y. Results : The mean age among 904 children was 4.5 years; 6.5% (59/904) were children <2 years old, 53.8% (486/904) were 2-5 years old, and 39.7% (359/904) were >5 years old; 52.0% (468/904) were male. N. meningitides was isolated from only 7 children attending 5 different day care centers and the overall carriage rate of N. meningitides was 0.8%. The detected serotypes of N. meningitides were serotype A (n=2), C (n=2), and Y (n=3). Conclusion : The carriage rate of N. meningitides among healthy children attending day care centers was very low in Korea and the detected serotypes were A, C, and Y.

  • PDF