• Title/Summary/Keyword: total RNA

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The Effects of Temperature, Starvation, and Kind of Foods on Growth, RNA/DNA Ratios and Protein Contents during the Early Life Stages of Olive Flounder (Paralichthys olivaceus)

  • Gwak Woo-Seok;Tanaka Masaru
    • Fisheries and Aquatic Sciences
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    • v.7 no.4
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    • pp.192-203
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    • 2004
  • The effects of temperatures, starvation, and kind of foods on growth, RNA/DNA ratio and protein contents during metamorphosis and early juvenile stage of olive flounder (Paralichthys olivaceus) were examined. During metamorphosis, warm-acclimated fish showed higher RNA and DNA content than those of the cold-acclimated fish, excepting H stage (28 DAH) at which the ratio was higher at cold temperature. RNA/DNA ratio during metamorphosis showed similar values at two temperatures tested. However, after 42 DAH warm-acclimated juveniles had higher DNA content compared with cold-acclimated fish, resulted in marked decreases in RNA/DNA ratios. Higher RNA content at H stage of cold-acclimated fish was consistent with an increase in protein content. Growth of fish rearing at warm temperature was higher than those of fish at cold temperature during all experiments. In starvation experiment, contents of DNA, RNA and protein significantly decreased. Even though there were no significant differences in total length (TL) and body weight between the live mysid-fed and artificial pellet-fed fish at 35 mm TL, both RNA/DNA and protein/DNA ratios of the former group was significantly higher than those of the latter due primarily to lower DNA content of the live mysid-fed group. The results from this study suggest that temperature, starvation and kind of foods should be considered when RNA/DNA ratio applied to assessing the cultured larval and juvenile fish condition.

Analysis of H3K4me3-ChIP-Seq and RNA-Seq data to understand the putative role of miRNAs and their target genes in breast cancer cell lines

  • Kotipalli, Aneesh;Banerjee, Ruma;Kasibhatla, Sunitha Manjari;Joshi, Rajendra
    • Genomics & Informatics
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    • v.19 no.2
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    • pp.17.1-17.13
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    • 2021
  • Breast cancer is one of the leading causes of cancer in women all over the world and accounts for ~25% of newly observed cancers in women. Epigenetic modifications influence differential expression of genes through non-coding RNA and play a crucial role in cancer regulation. In the present study, epigenetic regulation of gene expression by in-silico analysis of histone modifications using chromatin immunoprecipitation sequencing (ChIP-Seq) has been carried out. Histone modification data of H3K4me3 from one normal-like and four breast cancer cell lines were used to predict miRNA expression at the promoter level. Predicted miRNA promoters (based on ChIP-Seq) were used as a probe to identify gene targets. Five triple-negative breast cancer (TNBC)-specific miRNAs (miR153-1, miR4767, miR4487, miR6720, and miR-LET7I) were identified and corresponding 13 gene targets were predicted. Eight miRNA promoter peaks were predicted to be differentially expressed in at least three breast cancer cell lines (miR4512, miR6791, miR330, miR3180-3, miR6080, miR5787, miR6733, and miR3613). A total of 44 gene targets were identified based on the 3'-untranslated regions of downregulated mRNA genes that contain putative binding targets to these eight miRNAs. These include 17 and 15 genes in luminal-A type and TNBC respectively, that have been reported to be associated with breast cancer regulation. Of the remaining 12 genes, seven (A4GALT, C2ORF74, HRCT1, ZC4H2, ZNF512, ZNF655, and ZNF608) show similar relative expression profiles in large patient samples and other breast cancer cell lines thereby giving insight into predicted role of H3K4me3 mediated gene regulation via the miRNA-mRNA axis.

Cytokine Inductions and Intracellular Signal Profiles by Stimulation of dsRNA and SEB in the Macrophages and Epithelial Cells

  • Jun-Pyo Choi;Purevsuren Losol;Ghazal Ayoub;Mihong Ji;Sae-Hoon Kim;Sang-Heon Cho;Yoon-Seok Chang
    • IMMUNE NETWORK
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    • v.22 no.2
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    • pp.15.1-15.16
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    • 2022
  • Foreign molecules, including viruses and bacteria-derived toxins, can also induce airway inflammation. However, to the best of our knowledge, the roles of these molecules in the development of airway inflammation have not been fully elucidated. Herein, we investigated the precise role and synergistic effect of virus-mimicking double-stranded RNA (dsRNA) and staphylococcal enterotoxin B (SEB) in macrophages and epithelial cells. To identify cytokine expression profiles, both the THP-1-derived macrophages and BEAS-2B epithelial cells were stimulated with dsRNA or SEB. A total of 21 cytokines were evaluated in the culture supernatants. We observed that stimulation with dsRNA induced cytokine production in both cell types. However, cytokine production was not induced in SEB-stimulated epithelial cells, compared to the macrophages. The synergistic effect of dsRNA and SEB was evaluated observing cytokine level and intracellular phospho-signaling. Fifteen different types were detected in high-dose dsRNA-stimulated epithelial cells, and 12 distinct types were detected in macrophages; those found in macrophages lacked interferon production compared to the epithelial cells. Notably, a synergistic effect of cytokine induction by co-stimulation of dsRNA and SEB was observed mainly in epithelial cells, via activation of most intracellular phosphor-signaling. However, macrophages only showed an accumulative effect. This study showed that the type and severity of cytokine productions from the epithelium or macrophages could be affected by different intensities and a combination of dsRNA and SEB. Further studies with this approach may improve our understanding of the development and exacerbation of airway inflammation and asthma.

Studies on Freezing Tolerance of Mulberry, Morus species -Relation between Freezing Tolerance and Some Substances in Mulberry Branches- (뽕나무 내한성에 관한 연구 -특히 지조함유물질과 내한성과의 관계에 대하여-)

  • 김호락
    • Journal of Sericultural and Entomological Science
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    • v.22 no.1
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    • pp.7-25
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    • 1980
  • Some substances and freezing tolerance in the mulberry (Morus species) branch have been studied on the basis of varietal differences and harvesting times along with harvesting methods in autumn. The results obtained are summarized as follows; 1. The highest freezing tolerance was shown in the varieties of Yongcheon-chou, Jasan, Kang-weon No. 3 and Ichihei, the medium in Roso. Kairyonezumigaeshi, Yanagida and Kokuso No. 28, and the lowest in Ichinose, Mokuso, Kokuso No. 21 and Suweousang No. 3. 2. There was a signifiant negative correlation (r= -0.59*) between death atop percentage in the field and the temperature required to kill 50% of the mulberry buds (T$_{50}$) with the harvesting times and methods in autumn. Cold hardening occurred in the early through the end of September with the peak at the mid-september. During this period, leaf harvest decreased freezing tolerance with remarkable decrease due to picking all the leaves and leaving several leaves at the base of branch. Greater cold hardening was induced by leaving several leaves after topping. 3. Negative correlations were observed between freezing tolerance and the contents of soluble (r =-0.70*) and crude (r= -0.70*) protein. However, positive correlations were shown between freezing tolerance and total carbohydrate contents per crude (r=0.31*) and per soluble (r=0.71*) protein . There were also positive correlations between freezing tolerance and total sugar (r=0.67*) and RNA content (r=0.99**). No relationships of dry matter. fat. total carbohydrate and DNA contents were observed to the freezing tolerance. 4. Such sugars as raffinose. lactose, sucrose, glucose, fructose. arabinose. xylose. ribose (assumed) and rhamnose were detected in winter mulberry branch. Major sugars such as sucrose, glucose, and fructose were supposed to have higher relationship to the freezing tolerance than the other sugars. 5. Late harvesting increased RNA content except in the case of total leaf picking at mid-September. Leaf picking decreased RNA content. Some amount of RNA was, however, maintained by leaving several leaves after topping Leaving upper-middle leaves of a branch showed high RNA content. Leaving young leaves at the top and the overmatured leaves at the base showed low content. A positive correlation (r=0.51*) was noted between RNA content and freezing tolerance in the different harvesting methods.s.

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Turnover of $^{32}P$ and Total P in $^{32}P$-labeled Chlorella Cells in a Standard "Cold" Medium. ($^{32}P$-Labeled Chlorella의 정상배지에 있어서의 $^{32}P$ 및 Total P의 전환)

  • Lee, Yung-Nok;Chin, Pyung
    • Korean Journal of Microbiology
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    • v.4 no.1
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    • pp.14-20
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    • 1966
  • 1. Uniformly $^{32}P$-labeled Chlorella cells were further grown in a standard "cold" medium and aliquots of the algal cells were taken out at the beginning of, and at intervals during the culture, and subjected to analyze the contents of $^{32}$ P and total P in various fractions of the cell constituents. 2. When the $^{32}P$--labeled algae were grown in a normal "cold" medium, the P-contents in the fractions of DNA and protein increased. In the meantime the $^{32}P$- in acid-insoluble polyphosphate fraction decreased considerably, while that in RNA-polyphosphate complex significantly increased. 3. It was inferred that, under the experimental conditions of the present study, the phosphorus in polyphosphate seems to be transferred to RNA polyposphate complex and the phosphorus used in the synthesis of DNA and protein was, directly or indirectly, taken from those fractions above.ose fractions above.

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Vitellogenin and Its mRNA Induction by $Estradiol-17\beta$ in the Primary Culture of Hepatocytes in the Rainbow Trout, Oncorhynchus mykiss

  • Hwang Un-Gi
    • Fisheries and Aquatic Sciences
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    • v.4 no.4
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    • pp.186-191
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    • 2001
  • Vitellogenin (VTG) and VTG mRNA induction by $estradiol-17\beta\;(E_2)$ were examined in the primary cultures of hepatocyte in the rainbow trout. Hepatocytes were precultured for 2 days, then $E_2$ was added and cultured for another 5 days. Media and hepatocytes were then analyzed by electrophoresis and Northern blotting for VTG and VTG mRNA, respectively. The hepatocytes were formed a few aggregates within 5 days without further spreading to a monolayer. Cell viability and high DNA content were maintained during the incubation. The hepatocyte culture with E2 induced a weak VTG band at a molecular weight of 175kDa on Day 2 after $E_2$ addition. The relative amount of VTG was expressed in percentage of total protein concentrations. VTG was gradually increased as $1.9\%$ on Day 2, $6.3\%$ on Day 4 and $7.3\%$ on Day 5. VTG mRNA band was detected at about 6.6 kb in the culture with $E_2$ at day 1 of culture. The level of VTG mRNA expression linearly increased with time until Day 5 (r=0.97).

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Seasonal Change of RNA/DNA Ratio and Blood Characteristics of Black Sea Bream Acanthopagrus schlegeli Habituated in Geojae Costal Area, Kyungnam Province, Korea (경남 거제해역에 서식하는 감성돔(Acanthopagrus schlegeli)의 연간 RNA/DNA 및 혈액학적 특성 변화)

  • Kim, Su-Kyoung;Shim, Na-Young;Lee, Do-Hyun;Kim, Dae Hyun;Yoon, Seong-Jong
    • Journal of Environmental Science International
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    • v.22 no.1
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    • pp.37-45
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    • 2013
  • The monthly variations of blood characteristics and RNA/DNA of black sea bream, Acanthopagrus schlegeli, habituated in Geojae costal area were analysed to determine health condition of natural stocks in terms of gonad maturation and spawning season from March 2010 to February 2011. Spawning season determinated by gonadosomatic index is from June to August. RNA/DNA ratio of black sea bream muscle was strongly correlated with spawning season. During the gonad maturation RNA/DNA ratio in dorsal muscle tissue was decreased contrast to rapid increase during spawning season. Blood composition factors increased in terms of gonad maturation are aspartate aminotransferase, cholesterol, triglyceride, total protein, glucose, globulin, alkaline phosphatase and inorganic phosphate. Other blood factors increased during spawning season are alanine aminotransferase, blood urea nitrogen, uric acid and lactate dehydrogenase.

Regulation of Pharmacogene Expression by microRNA in The Cancer Genome Atlas (TCGA) Research Network

  • Han, Nayoung;Song, Yun-Kyoung;Burckart, Gilbert J.;Ji, Eunhee;Kim, In-Wha;Oh, Jung Mi
    • Biomolecules & Therapeutics
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    • v.25 no.5
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    • pp.482-489
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    • 2017
  • Individual differences in drug responses are associated with genetic and epigenetic variability of pharmacogene expression. We aimed to identify the relevant miRNAs which regulate pharmacogenes associated with drug responses. The miRNA and mRNA expression profiles derived from data for normal and solid tumor tissues in The Cancer Genome Atlas (TCGA) Research Network. Predicted miRNAs targeted to pharmacogenes were identified using publicly available databases. A total of 95 pharmacogenes were selected from cholangiocarcinoma and colon adenocarcinoma, as well as kidney renal clear cell, liver hepatocellular, and lung squamous cell carcinomas. Through the integration analyses of miRNA and mRNA, 35 miRNAs were found to negatively correlate with mRNA expression levels of 16 pharmacogenes in normal bile duct, liver, colon, and lung tissues (p<0.05). Additionally, 36 miRNAs were related to differential expression of 32 pharmacogene mRNAs in those normal and tumorigenic tissues (p<0.05). These results indicate that changes in expression levels of miRNAs targeted to pharmacogenes in normal and tumor tissues may play a role in determining individual variations in drug response.

Characterization of X-linked RNA Transcripts in Matured Bovine Spermatozoa

  • Jeon, Byeong-Gyun;Kumar, B. Mohana;Rho, Gyu-Jin
    • Reproductive and Developmental Biology
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    • v.35 no.3
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    • pp.209-214
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    • 2011
  • Although the function and utility of RNA transcripts derived from matured spermatozoa remains unclear, they might play important roles in the establishment of a paternal genome and subsequently embryo development. Herein, we investigated the expression of X-chromosome linked RNA transcripts in matured bovine spermatozoa. The total RNA was extracted from the matured spermatozoa, and then converted to cDNA. Autosomal genes (ACT-${\beta}$ and H-2A) and X-chromosome linked genes (ANT3, HPRT, MeCP2, RPS4X, XIAP, XIST and ZFX) were analyzed for the characterization of X-chromosome linked RNA transcripts and compared to female fibroblasts by RT-PCR. The transcripts of autosomal genes (ACT-${\beta}$ and H2A) and X-chromosome linked genes (ANT3, HPRT, MeCP2, RPS4X and ZFX) were not detected in spermatozoa. However, XIAP (X-linked inhibitor of apoptosis protein) and XIST (X-chromosome inactive-specific transcript, a kind of paternal imprinted gene) transcripts were detected in spermatozoa, and relative levels of XIAP and XIST transcripts were similar and 0.5-fold lower when compared to female fibroblasts, respectively. Based on the findings, it is summarized that the presence of RNA transcripts of XIAP and XIST in the isolated spermatozoa may imply their role in inhibition of apoptosis and induction of X-chromosome inactivation in embryo development.

Induction of ER-stress by Heat Shock in the Thyrocytes

  • Kwon, Ki-Sang;Kwon, O-Yu;Yang, Young-Mo
    • Biomedical Science Letters
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    • v.12 no.4
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    • pp.435-438
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    • 2006
  • In eukaryotes, ER stress induces UPR (unfolded protein response) via IRE1 activation which sends a molecular signal for XBP1 mRNA splicing in the cytosol. During this mRNA splicing, 23 nt removed in which contains PstI site and then resulting XBP1 product is not digested with PstI restriction enzyme. In this study, using this XBP1 mRNA splicing mechanism, the effect of heat shock on thyrocytes is studied, because heat shock response in the thyrocytes needs more study to understand thyroid physiology under alternative environments. ER inducible drugs (tunicamycin, DTT, $Ca^{2+}$ ionopore A23187, BFA) induce ER stress in the thyrocytes. From 3 hours after heat shock, ER stress is induced and which is reversible when heat shock is without. While $Ca^{2+}$ ionopore A23187 is reversible from ER stress by washing out the drug, thapsigagin is irreversible. Other ER inducible drugs are not so sensitive to ER stress repairing. XBP1 mRNA splicing in a cell is very available method to detect ER stress. It needs only a small quantity of total RNA and processing also very easy.

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