• Title/Summary/Keyword: single gene analysis

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Obestatin is present in saliva: alterations in obestatin and ghrelin levels of saliva and serum in ischemic heart disease

  • Ozbay, Yilmaz;Aydin, Suleyman;Dagli, A. Ferda;Akbulut, Mehmet;Dagli, Necati;Kilic, Nermin;Rahman, Ali;Sahin, Ibrahim;Polat, Veli;Ozercan, H. Ibrahim;Arslan, Nadi;Sensoy, Dogan
    • BMB Reports
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    • v.41 no.1
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    • pp.55-61
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    • 2008
  • Ghrelin and obestatin are a single gene products and are a multiple functional peptides that regulates energy homeostasis, and food intake. In the present work, we studied the secretion of ghrelin and its co-secreted peptide obestatin in 44 patients with ischemic heart disease with that of 27 healthy matched controls. Here we first conducted using an immunohistochemistry assay to screen whether human salivary glands have any obestatin immunoreactivity. Then, serum and saliva obestatin and acylated ghrelin levels were determined by using Radioimmunoassay. Our immunohistochemical analysis demonstrated that obestatin was localized in the striated and excretory duct of human salivary gland. We also report for the first time that obestatin, like ghrelin, is present in human salivary gland and saliva. No evidence of the role of obestatin or ghrelin saliva levels in the context of ischemic heart disease was found. Salivary ghrelin and obestatin levels are correlated in controls with the blood levels. Determination of salivary values could represent a non-invasive alternative to serum ones that can be useful in clinical practice.

Association of GRIA1 polymorphisms with ovarian response to human menopausal gonadotropin in Iranian women

  • Golestanpour, Hossein;Javadi, Gholamreza;Sheikhha, Mohammad Hasan
    • Clinical and Experimental Reproductive Medicine
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    • v.47 no.3
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    • pp.207-212
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    • 2020
  • Objective: Glutamate ionotropic receptor AMPA type subunit 1 (GRIA1) is a subunit of a ligand-gated ion channel that regulates the secretion of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) by controlling the release of gonadotropin-releasing hormone. Few studies have investigated the association between the GRIA1 gene and human infertility. This study evaluated the association of the GRIA1 rs548294 C > T and rs2195450 G > A polymorphisms with the ovarian response to human menopausal gonadotropin (HMG) in Iranian women. Methods: One hundred women with histories of at least 1 year of infertility were included. On the second day of menstruation, patients were injected with HMG; on the third day, blood samples were collected. After hormonal analysis, the GRIA1 rs548294 C > T and rs2195450 G > A genotypes of samples were identified via the restriction fragment length polymorphism method, and on day 9, the number of follicles was assessed via ultrasound. Results: For the GRIA1 rs548294 C > T and rs2195450 G > A single nucleotide polymorphisms, the subjects with CT and GG genotypes, respectively, displayed the highest mean FSH level, LH level, and number of follicles on day 9 of the menstrual cycle (p< 0.05). Significant positive correlations were observed between LH and FSH (p< 0.01), LH and follicle count (p< 0.01), FSH and age (p< 0.05), follicle count and age (p= 0.048), and FSH and follicle count (p< 0.01). Conclusion: This study showed a significant relationship between GRIA1 polymorphisms and ovarian response to the induction of ovulation. Therefore, determining patients' GRIA1 genotype may be useful for improving treatment and prescribing suitable doses of ovulation-stimulating drugs.

Effects of Gleditsiae Spina(GS) on the Polycystic Ovary Induced by Estradiol Valerate in Rats (조각자(皂角刺)가 estradiol valerate로 유발된 백서(白鼠)의 다낭성 난소에 미치는 영향)

  • Gu, Hee-Jun;Cho, Sung-Hee
    • The Journal of Korean Obstetrics and Gynecology
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    • v.23 no.2
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    • pp.71-84
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    • 2010
  • Purpose: In the theory of traditional medicine, Glenditsia spina(GS) can resolve carbuncle, relive swelling, dispel wind and destroy parasites. This study was designed to investigate the effects of GS on gene expression of ovarian tissue in polycystic ovary syndrome(PCOS) rats. Methods: In this experiment, female rats injected with a single dose of 2 mg estradiol valerate(EV) and GS was given for 5 weeks. The genetic profile for the effects on ovarian tissue in PCOS rats was measured using microarray technique, and the functional analysis on these genes was conducted. Results: 985 genes were increased in control and restored to normal level in GS group. (B), 733 genes were decreased in control group and restored to normal level in GS group. (F). Metabolic pathways related in B group genes were Graft-versus-host disease, Allograft rejection, Autoimmune thyroid disease, Cytokine-cytokine receptor interaction, Small cell lung cancer, Type I diabetes mellitus. Metabolic pathways related in F group genes were Antigen processing and present, Adipocytokine signalling pathway, Focal adhesion, ECM-receptor interaction, Pancreatic cancer, Notch signalling pathway, Tight junction. The network of total protein interactions was measured using cytoscape program, and some key molecules, such as c-Fos, c-Myc, ABL1 related in B group, MAPK8, RASA1, CALR related in F group that can be used for elucidation of therapeutical mechanism of medicine in future were identified. Conclusion: These results suggest possibility of GS as anti-cancer and anti-hyperplasia drug in PCOS. In addition, the present author also suggests that related mechanisms are involved in suppression of proto-oncogene such as c-Fos, c-Myc and ABL1, and in regulation of cell cycle such as RASA1.

Identification of DNA Variations Using AFLP and SSR Markers in Soybean Somaclonal Variants

  • Lee, Suk-Ha;Jung, Hyun-Soo;Kyujung Van;Kim, Moon-Young
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.1
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    • pp.69-72
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    • 2004
  • Somaclonal variation, defined as phenotypic and genetic variations among regenerated plants from a parental plant, could be caused by changes in chromosome structure, single gene mutation, cytoplasm genetic mutation, insertion of transposable elements, and DNA methylation during plant regeneration. The objective of this study was to evaluate DNA variations among somaclonal variants from the cotyledonary node culture in soybean. A total of 61 soybean somaclones including seven $\textrm{R}_1$ lines and seven $\textrm{R}_2$ lines from Iksannamulkong as well as 27 $\textrm{R}_1$ lines and 20 $\textrm{R}_2$ lines from Jinju 1 were regenerated by organogenesis from the soybean cotyledonary node culture system. Field evaluation revealed no phenotypic difference in major agronomic traits between somaclonal variants and their wild types. AFLP and SSR analyses were performed to detect variations at the DNA level among somaclonal variants of two varieties. Based on AFLP analysis using 36 primer sets, 17 of 892 bands were polymorphic between Iksannamulkong and its somaclonal variants and 11 of 887 bands were polymorphic between Jinju 1 and its somaclonal variants, indicating the presence of DNA sequence change during plant regeneration. Using 36 SSR markers, two polymorphic SSR markers were detected between Iksannamulkong and its somaclonal variants. Sequence comparison amplified with the primers flanking Satt545 showed four additional stretches of ATT repeat in the variant. This suggests that variation at the DNA level between somaclonal variants and their wild types could provide basis for inducing mutation via plant regeneration and broadening crop genetic diversity.

Isolation, Physicochemical Properties and Toxicities of Territrem A' and B'

  • Ling, Kuo Huang;Peng, Fu Chuo;Chen, Bai Jiun;Wang, Yu;Lee, Gene Hsiang
    • Korean Journal of Pharmacognosy
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    • v.17 no.2
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    • pp.153-160
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    • 1986
  • We have isolated two new metabolites of territrem, designated as territrem $A'\;(TRA';\;C_{28}H_{30}O_{10})$ and $B'\;(TRB';\; C_{29}H_{34}O_{10})$ from chloroform extract of rice culture of Aspergillus terreus 23-1, using the same isolation procedure as that for territrem A, B and C(TRA, TRB, TRC). The present isolation procedure gave about 5 mg of TRA' and 10 mg of TRB' from 4 kg of rice culture per batch. Analysis of the high resolution mass spectrum showed that the molecular composition of TRA' and TRB' are $C_{28}H_{30}O_{10}$ and $C_{29}H_{34}O_{10}$ respectively, Some results of physicochemical properties were presented in this paper. Single crystal X-ray diffractometry of TRB' showed that the three dimensional structure of TRB' has not changed significantly from that of $TRB\;(C_{29}H_{34}O_9)$, except for the insertion of one oxygen atom into TRB to make additional pyran in the E-ring. It is also suggested that the aromatic moiety of TRA' is similar to that of $TRA\;(C_{28}H_{30}O_9)$ and the rest non-aromatic portions resemble to those of TRB'. The tremorgenic activity, lethality and inhibitory effect on acetylcholine esterase of TRA' and TRB' are greatly reduced comparing to that of TRA and TRB.

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Identification of Brucella melitensis isolates originating from Mongolia and diagnostic real-time PCR evaluation using a specific SNP (몽골 유래 Brucella melitensis 동정 및 특이 SNP를 이용한 real-time PCR법에 의한 진단 평가)

  • Kang, Sung-Il;Kim, Ji-Yeon;Kim, Suk Mi;Lee, Jin Ju;Sung, So-Ra;Kim, Yeon-Hee;Jung, Suk Chan;Her, Moon
    • Korean Journal of Veterinary Research
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    • v.55 no.2
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    • pp.105-110
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    • 2015
  • A real-time PCR assay using hybridization probe (HybProbe) has been developed to detect Brucella (B.) melitensis strains. The primer and HybProbe sets were designed based on the gap gene of chromosome I with a specific single nucleotide polymorphism of B. melitensis. Specificity of the assay was confirmed by comparison to reference Brucella species and other related strains. In the melting curve analysis, B. melitensis generated a peak at $67^{\circ}C$ unlike those for other Brucella species observed at $61^{\circ}C$. Sensitivity of the assay for B. melitensis ranged from 20 ng to 200 fg of genomic DNA. The ability to identify 94 Mongolian B. melitensis isolates using the real-time PCR assay was identical to that of classical biotyping methods and differential multiplex PCR. These data showed that this new molecular technique is a simple and quick method for detecting B. melitensis, which will be important for the control and prevention of brucellosis.

Therapeutic Potential of an Anti-diabetic Drug, Metformin: Alteration of miRNA expression in Prostate Cancer Cells

  • Avci, Cigir Biray;Harman, Ece;Dodurga, Yavuz;Susluer, Sunde Yilmaz;Gunduz, Cumhur
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.2
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    • pp.765-768
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    • 2013
  • Background and Aims: Prostate cancer is the most commonly diagnosed cancer in males in many populations. Metformin is the most widely used anti-diabetic drug in the world, and there is increasing evidence of a potential efficacy of this agent as an anti-cancer drug. Metformin inhibits the proliferation of a range of cancer cells including prostate, colon, breast, ovarian, and glioma lines. MicroRNAs (miRNAs) are a class of small, non-coding, single-stranded RNAs that downregulate gene expression. We aimed to evaluate the effects of metformin treatment on changes in miRNA expression in PC-3 cells, and possible associations with biological behaviour. Materials and Methods: Average cell viability and cytotoxic effects of metformin were investigated at 24 hour intervals for three days using the xCELLigence system. The $IC_{50}$ dose of metformin in the PC-3 cells was found to be 5 mM. RNA samples were used for analysis using custom multi-species microarrays containing 1209 probes covering 1221 human mature microRNAs present in miRBase 16.0 database. Results: Among the human miRNAs investigated by the arrays, 10 miRNAs were up-regulated and 12 miRNAs were down-regulated in the metformin-treated group as compared to the control group. In conclusion, expression changes in miRNAs of miR-146a, miR-100, miR-425, miR-193a-3p and, miR-106b in metformin-treated cells may be important. This study may emphasize a new role of metformin on the regulation of miRNAs in prostate cancer.

Functional Expression and Characterization of Recombinant NADPH-P450 Reductase from Malassezia globosa

  • Lee, Hwa-Youn;Park, Hyoung-Goo;Lim, Young-Ran;Lee, Im-Soon;Kim, Beom-Joon;Seong, Cheul-Hun;Chun, Young-Jin;Kim, Dong-Hak
    • Journal of Microbiology and Biotechnology
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    • v.22 no.1
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    • pp.141-146
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    • 2012
  • Malassezia globosa is a common pathogenic fungus that causes skin diseases including dandruff and seborrheic dermatitis in humans. Analysis of its genome identified a gene (MGL_1677) coding for a putative NADPH-P450 reductase (NPR) to support the fungal cytochrome P450 enzymes. The heterologously expressed recombinant M. globosa NPR protein was purified, and its functional features were characterized. The purified protein generated a single band on SDS-PAGE at 80.74 kDa and had an absorption maximum at 452 nm, indicating its possible function as an oxidized flavin cofactor. It evidenced NADPH-dependent reducing activity for cytochrome c or nitroblue tetrazolium. Human P450 1A2 and 2A6 were able to successfully catalyze the O-deethylation of 7-ethoxyresorufin and the 7-hydroxylation of coumarin, respectively, with the support of the purified NPR. These results demonstrate that purified NPR is an orthologous reductase protein that supports cytochrome P450 enzymes in M. globosa.

Agronomic Characteristics of Rice Recombinant Inbred Lines (RILs) Developed from a Cross of Nonganbyeo and BG 279

  • Lee, Jeom-Ho;Kim, Nam-Soo;Cho, Youn-Sang;Song, Moon-Tae;Hwang, Hung-Goo;Moon, Huhn-Pal
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.44 no.1
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    • pp.86-94
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    • 1999
  • A set of rice recombinant inbred lines was developed from a cross between a Tongil type variety, Nonganbyeo, and an indica variety, BG276, by the single seed descent method. The number of the lines in the population was 272. All the agronomic characters studied except ADV (alkali-digestion value) showed continuous variation among the RILs, implying that their inheritance mode should be quantitative. The patterns of the variation in the RILs were either normal or skewed distribution. ADVs of RILs were segregated into two groups with 1:1 ratio, indicating that ADVs in this KIL population might be controlled by one major gene. Transgressive variations were also observed in all characters. Heritability values of the characters varied from 0.488 in brown/rough rice ratio to 0.895 in alkali-digestion value. In the analysis of genotypic and phenotypic correlations, the character of yield was positively correlated with 8 different agronomic characters. The number of panicles per hill was negatively correlated with culm length, panicle length, and number of spikelets per panicle. Grain length was positively correlated with grain width, grain thickness, grain length/width ratio, white belly, ADV, and amylose. However, grain length/width ratio was negatively correlated with grain width. White core was also negatively correlated with white belly and ADV.

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First detection of avian polyomavirus by PCR from Alexandrine Parakeet (Psittacula eupatria) in Korea (대본청 앵무(Psittacula eupatria )로부터 PCR에 의한 avian polyomavirus 최초 검출)

  • Kim, Hee-Jung;Lee, Sun-Rock;Park, Choi-Kyu
    • Korean Journal of Veterinary Service
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    • v.37 no.3
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    • pp.213-218
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    • 2014
  • In early April 2014, a month-old Alexandrine Paraqeet (Psittacula eupatria) that was raised in a domestic aviary located in Gyungju-si, Korea was suddenly died and submitted to Animal Disease Intervention Center, Kyungpook National University in order to diagnose the causative agent. In post-mortem examination, the bird had abnormally developed feathers on the neck and abdomen region and subcutaneous hemorrhages on the neck and cheek adjacent to the beak. At necropsy, the bird had hemorrhage on the muscle of the femoral region, ascites, multi-focal hemorrhages on the epicardium, and diffuse hemorrhages on the sub-serosa of proventriculus and gizzard, suggesting typical avian polyomavirus (APV) infection. The partial large tumor (T) antigen gene of APV was detected by PCR from tissues of the heart, lung, liver, kidney, proventriculus and feathers of the APV-suspected birds. However, other pathogenic virus-specific nucleic acid common with psittacine birds such as avian bornavirus, psittacine beak and feather disease virus and psittacid herpesvirus were not detected from the mixed tissue samples of the bird, indicating this case is due to single infection of APV. Nucleotide sequence analysis of the partially amplified large T antigen DNA was confirmed to have 99~100% homology with that of the previously reported APV strains. This case report describes the first detection of APV in Alexandrine Paraqeet in Korea.