• Title/Summary/Keyword: Housekeeping gene

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KRT13, FAIM2 and CYP2W1 mRNA Expression in Oral Squamous Cell Carcinoma Patients with Risk Habits

  • Hartanto, Firstine Kelsi;Karen-Ng, Lee Peng;Vincent-Chong, Vui King;Ismail, Siti Mazlipah;Mustafa, Wan Mahadzir Wan;Abraham, Mannil Thomas;Tay, Keng Kiong;Zain, Rosnah Binti
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.3
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    • pp.953-958
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    • 2015
  • Background: Expression of KRT13, FAIM2 and CYP2W1 appears to be influenced by risk habits, thus exploring the associations of these genes in oral squamous cell cancer (OSCC) with risk habits, clinico-pathological parameters and patient survival may be beneficial in identifying relevant biomarkers with different oncogenic pathways. Materials and Methods: cDNAs from 41 OSCC samples with and without risk habits were included in this study. Quantitative real-time PCR was used to analyze KRT13, FAIM2 and CYP2W1 in OSCC. The housekeeping gene (GAPDH) was used as an endogenous control. Results: Of the 41 OSCC samples, KRT13 was down-regulated in 40 samples (97.6%), while FAIM2 and CYP2W1 were down-regulated in 61.0% and 48.8%, respectively. Overall, there were no associations between KRT13, FAIM2 and CYP2W1 expression with risk habits, selected socio-demographic and clinico-pathological parameters and patient survival. Conclusions: Although this study was unable to show significance, there were some tendencies in the associations of KRT13, FAIM2 and CYP2W1 expression in OSCC with selected clinic-pathological parameters and survival.

Xanthomonas euvesicatoria Causes Bacterial Spot Disease on Pepper Plant in Korea

  • Kyeon, Min-Seong;Son, Soo-Hyeong;Noh, Young-Hee;Kim, Yong-Eon;Lee, Hyok-In;Cha, Jae-Soon
    • The Plant Pathology Journal
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    • v.32 no.5
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    • pp.431-440
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    • 2016
  • In 2004, bacterial spot-causing xanthomonads (BSX) were reclassified into 4 species-Xanthomonas euvesicatoria, X. vesicatoria, X. perforans, and X. gardneri. Bacterial spot disease on pepper plant in Korea is known to be caused by both X. axonopodis pv. vesicatoria and X. vesicatoria. Here, we reidentified the pathogen causing bacterial spots on pepper plant based on the new classification. Accordingly, 72 pathogenic isolates were obtained from the lesions on pepper plants at 42 different locations. All isolates were negative for pectolytic activity. Five isolates were positive for amylolytic activity. All of the Korean pepper isolates had a 32 kDa-protein unique to X. euvesicatoria and had the same band pattern of the rpoB gene as that of X. euvesicatoria and X. perforans as indicated by PCR-restriction fragment length polymorphism analysis. A phylogenetic tree of 16S rDNA sequences showed that all of the Korean pepper plant isolates fit into the same group as did all the reference strains of X. euvesicatoria and X. perforans. A phylogenetic tree of the nucleotide sequences of 3 housekeeping genes-gapA, gyrB, and lepA showed that all of the Korean pepper plant isolates fit into the same group as did all of the references strains of X. euvesicatoria. Based on the phenotypic and genotypic characteristics, we identified the pathogen as X. euvesicatoria. Neither X. vesicatoria, the known pathogen of pepper bacterial spot, nor X. perforans, the known pathogen of tomato plant, was isolated. Thus, we suggest that the pathogen causing bacterial spot disease of pepper plants in Korea is X. euvesicatoria.

Zinc Status Assessment by Analysis of Mononuclear Cell Metallothionein mRNA Using Competitive-Reverse Transcriptase-Polymerase Chain Reaction

  • Lee, Soo-Lim;Yoon, Jin-Sook;Kwon, Chong-Suk;Beattie, John H.;Kwun, In-Sook
    • Preventive Nutrition and Food Science
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    • v.9 no.3
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    • pp.276-282
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    • 2004
  • Marginal Zn deficiency is prevalent through the world and yet human zinc status has not been properly assessed due to the lack of a reliable diagnostic indicator. One potential possibility for zinc status assessment using Zn-binding protein, metallothionein (MT)-mRNA, has been proposed. The purpose of the present study was aimed to show whether measurement of mononuclear cell (MNC) MT mRNA, using a competitive-reverse transcriptase-polymerase chain reaction (competitive-RT-PCR) assay, could indicate zinc status in human subjects. In this study, MNC MT-mRNA expression was measured using a competitive-RT-PCR to compare before and after 14 days of zinc supplementation (50 mg Zn/das zinc gluconate). RT-PCR oligonucleotide primers which were designed to amplify both a 278 bp segment of the human MT-2A cDNA and a 198 bp mutant competitor cDNA template from MNCs, were prepared. MT-2A mRNA was normalized by reference to the housekeeping gene, $\beta$-actin, mRNA for which was also measured by competitive-RT-PCR. There was considerable inter-individual variation in MT-mRNA concentration and yet, the mean MT-2A mRNA level increased 4.7-fold after Zn supplementation, as compared to before Zn supplementation. This MT-2A mRNA level was shown as the same pattern and, even more sensitive assay, compared to the conventional plasma and red blood cells (RBCs) Zn assessment in which plasma and RBCs zinc levels increased 2.3- and 1.2-fold, respectively (p<0.05). We suggest that MT competitive-RT-PCR can be a useful assessment tool for evaluating human zinc status.

Insights into Systems for Iron-Sulfur Cluster Biosynthesis in Acidophilic Microorganisms

  • Myriam, Perez;Braulio, Paillavil;Javiera, Rivera-Araya;Claudia, Munoz-Villagran;Omar, Orellana;Renato, Chavez;Gloria, Levican
    • Journal of Microbiology and Biotechnology
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    • v.32 no.9
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    • pp.1110-1119
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    • 2022
  • Fe-S clusters are versatile and essential cofactors that participate in multiple and fundamental biological processes. In Escherichia coli, the biogenesis of these cofactors requires either the housekeeping Isc pathway, or the stress-induced Suf pathway which plays a general role under conditions of oxidative stress or iron limitation. In the present work, the Fe-S cluster assembly Isc and Suf systems of acidophilic Bacteria and Archaea, which thrive in highly oxidative environments, were studied. This analysis revealed that acidophilic microorganisms have a complete set of genes encoding for a single system (either Suf or Isc). In acidophilic Proteobacteria and Nitrospirae, a complete set of isc genes (iscRSUAX-hscBA-fdx), but not genes coding for the Suf system, was detected. The activity of the Isc system was studied in Leptospirillum sp. CF-1 (Nitrospirae). RT-PCR experiments showed that eight candidate genes were co-transcribed and conform the isc operon in this strain. Additionally, RT-qPCR assays showed that the expression of the iscS gene was significantly up-regulated in cells exposed to oxidative stress imposed by 260 mM Fe2(SO4)3 for 1 h or iron starvation for 3 h. The activity of cysteine desulfurase (IscS) in CF-1 cell extracts was also upregulated under such conditions. Thus, the Isc system from Leptospirillum sp. CF-1 seems to play an active role in stressful environments. These results contribute to a better understanding of the distribution and role of Fe-S cluster protein biogenesis systems in organisms that thrive in extreme environmental conditions.

Identification of heat shock protein70-2 and protamine-1 mRNA, proteins, and analyses of their association with fertility using frozen-thawed sperm in Madura bulls

  • Zulfi Nur Amrina Rosyada;Berlin Pandapotan Pardede;Ekayanti Mulyawati Kaiin;Ligaya I.T.A Tumbelaka;Dedy Duryadi Solihin;Bambang Purwantara;Mokhamad Fakhrul Ulum
    • Animal Bioscience
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    • v.36 no.12
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    • pp.1796-1805
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    • 2023
  • Objective: This study aims to identify heat shock protein70-2 (HSP70-2) and protamine-1 (PRM1) mRNA and protein in Madura bull sperm and demonstrate their relation as bull fertility biomarkers. Methods: The Madura bull fertility rates were grouped based on the percentage of first service conception rate (%FSCR) as high fertility (HF) (79.04%; n = 4), and low fertility (LF) (65.84%; n = 4). mRNA of HSP70-2 and PRM1 with peptidylprolyl isomerase A (PPIA) as a housekeeping gene were determined by quantitative real-time polymerase chain reaction, while enzyme-linked immunoassay was used to measure protein abundance. In the post-thawed semen samples, sperm motility, viability, acrosome integrity, and sperm DNA fragmentation index were analyzed. Data analysis was performed on the measured parameters of semen quality, relative mRNA expression, and protein abundance of HSP70-2 and PRM1, among the bulls with various fertility levels (HF and LF) in a one-way analysis of variance analysis. The Pearson correlation was used to analyze the relationship between semen quality, mRNA, proteins, and fertility rate. Results: Relative mRNA expression and protein abundance of HSP70-2 and PRM1 were detected and were found to be highly expressed in bulls with HF (p<0.05) and were associated with several parameters of semen quality. Conclusion: HSP70-2 and PRM1 mRNA and protein molecules have great potential to serve as molecular markers for determining bull fertility.

Effect of adrenalectomy on gene expression of adrenoceptor subtypes in the hypothalamic paraventricular nucleus

  • Kam, Kyung-Yoon;Shin, Seung Yub;Han, Seong Kyu;Li, Long Hua;Chong, Wonee;Baek, Dae Hyun;Lee, So Yeong;Ryu, Pan Dong
    • Korean Journal of Veterinary Research
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    • v.44 no.2
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    • pp.207-215
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    • 2004
  • It is well known that the hypothalamic-pituitary-adrenocortical (HPA) axis is under the negative feedback control of adrenal corticosteroids. Previous studies have suggested that glucocorticoids can regulate neuroendocrine cells in the paraventricular nucleus (PVN) by modulating catecholaminergic transmission, a major excitatory modulator of the HPA axis at the hypothalamic level. But, the effects of corticosteroids on the expression of adrenoceptor subtypes are not fully understood. In this work, we examined mRNA levels of six adrenoceptor subtypes (${\alpha}_{1A}$, ${\alpha}_{1B}$, ${\alpha}_{2A}$, ${\alpha}_{2B}$, ${\beta}_1$ and ${\beta}_2$) in the PVN of normal and adrenalectomized (ADX) rats. Total RNA ($2.5{\mu}g$) was extracted from PVN micropunches of brain slices ($500{\mu}m$) and analyzed by reverse transcription-polymerase chain reaction (RT-PCR). The levels of corticotropin-releasing hormone (CRH) mRNA were increased in the ADX rats relative to normal rats, indicating that the PVN had been liberated from the negative feedback of corticosteroids. Among the six adrenoceptor subtypes examined, mRNA levels for ${\alpha}_{1B}$- and ${\beta}_1$-adrenoceptors were increased, but the level for ${\beta}_2$-adrenoceptors was decreased in the ADX rats. The mRNA levels for the other three subtypes and for the general and neuronal specific housekeeping genes, glyceroaldehyde-3-phosphate dehydrogenase (GAPDH) and N-enolase, respectively, were not changed in the ADX rats. In conclusion, the results indicate that adrenal steroids selectively regulate the gene expression of adrenoceptor subtypes in the PVN.

Bacterial Spot Disease of Green Pumpkin by Pseudomonas syringae pv. syringae (Pseudomonas syringae pv. syringae에 의한 애호박 세균점무늬병)

  • Park, Kyoung-Soo;Kim, Young-Tak;Kim, Hye-Seong;Lee, Ji-Hye;Lee, Hyok-In;Cha, Jae-Soon
    • Research in Plant Disease
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    • v.22 no.3
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    • pp.158-167
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    • 2016
  • A pathogen that causes a new disease on green pumpkin in the nursery and the field was characterized and identified. Symptoms of the disease on green pumpkin were water soaking lesions and spots with strong yellow halo on leaf, brown lesion on flower, and yellow spot on fruit. The bacterial isolates from the leaf spot were pathogenic on the 8 curcubitaceae crop plants, green pumpkin, figleaf gourd, wax gourd, young pumpkin, zucchini, cucumber, melon, and oriental melon, whereas they did not cause the disease on sweet pumpkin and watermelon. They were Gram-negative, rod shape with polar flagella, fluorescent on King's B agar and LOPAT group 1a by LOPAT test. Their Biolog substrate utilization patterns were similar to Pseudomonas syringae pv. syringae's in Biolog database. Phylogenetic trees with 16S rRNA gene sequences and multilocus sequence typing (MLST) with nucleotide sequences of 4 housekeeping genes, gapA, gltA, gyrB, rpoD and those of P. syringae complex strains in the Plant Associated and Environmental Microbes Database (PAMDB) showed that the green pumpkin isolates formed in the same clade with P. syringae pv. syringae strains. The clade in MLST tree was in the genomospecies 1 group. The phenotypic and genotypic characteristics suggested that the isolates from green pumpkin lesion were P. syringae pv. syringae.