• Title/Summary/Keyword: Systems Biology

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Visualizing Live Chromatin Dynamics through CRISPR-Based Imaging Techniques

  • Chaudhary, Narendra;Im, Jae-Kyeong;Nho, Si-Hyeong;Kim, Hajin
    • Molecules and Cells
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    • v.44 no.9
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    • pp.627-636
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    • 2021
  • The three-dimensional organization of chromatin and its time-dependent changes greatly affect virtually every cellular function, especially DNA replication, genome maintenance, transcription regulation, and cell differentiation. Sequencing-based techniques such as ChIP-seq, ATAC-seq, and Hi-C provide abundant information on how genomic elements are coupled with regulatory proteins and functionally organized into hierarchical domains through their interactions. However, visualizing the time-dependent changes of such organization in individual cells remains challenging. Recent developments of CRISPR systems for site-specific fluorescent labeling of genomic loci have provided promising strategies for visualizing chromatin dynamics in live cells. However, there are several limiting factors, including background signals, off-target binding of CRISPR, and rapid photobleaching of the fluorophores, requiring a large number of target-bound CRISPR complexes to reliably distinguish the target-specific foci from the background. Various modifications have been engineered into the CRISPR system to enhance the signal-to-background ratio and signal longevity to detect target foci more reliably and efficiently, and to reduce the required target size. In this review, we comprehensively compare the performances of recently developed CRISPR designs for improved visualization of genomic loci in terms of the reliability of target detection, the ability to detect small repeat loci, and the allowed time of live tracking. Longer observation of genomic loci allows the detailed identification of the dynamic characteristics of chromatin. The diffusion properties of chromatin found in recent studies are reviewed, which provide suggestions for the underlying biological processes.

Risk factors of African swine fever virus in suspected infected pigs in smallholder farming systems in South-Kivu province, Democratic Republic of Congo

  • Bisimwa, Patrick N.;Dione, Michel;Basengere, Bisimwa;Mushagalusa, Ciza Arsene;Steinaa, Lucilla;Ongus, Juliette
    • Journal of Veterinary Science
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    • v.22 no.3
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    • pp.35.1-35.13
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    • 2021
  • Background: African swine fever (ASF) is an infectious viral disease of domestic pigs that presents as a hemorrhagic fever, and for which no effective vaccine is available. The disease has a serious negative social and economic impact on pig keepers. There is limited information on the potential risk factors responsible for the spread of ASF in South Kivu. Objective: The aim of this study was to determine the potential risk factors associated with ASF infection in suspected ASF virus (ASFV)-infected pigs. Methods: We sampled whole blood from 391 pigs. Additionally, 300 pig farmers were interviewed using a structured questionnaire. Viral DNA was detected by using the real-time polymerase chain reaction technique. Results: The majority of pigs sampled, 78% (95% confidence interval [CI], 74.4-82.6), were of local breeds. Over half, 60.4% (95% CI, 55.5-65.2), were female, and most of them, 90.5% (95% CI, 87.6-93.4), were adult pigs (> 1 year old). Viral DNA was detected in 72 of the 391 sampled pigs, indicating an overall infection rate of 18.4% (95% CI, 14.5-22.4). Multivariable logistic regression analysis revealed several risk factors positively associated with ASFV infection: feeding with swill in pen (odds ratio [OR], 3.8; 95% CI, 2.12-6.77); mixed ages of pigs in the same pen (OR, 3.3; 95% CI, 1.99-5.57); introduction of new animals to the farm (OR, 5.4; 95% CI, 1.91-15.28). The risk factors that were negatively (protective) correlated with ASFV positivity were the presence of male animals and the use of an in-pen breeding system. Conclusion: Local pig farmers should be encouraged to adopt proper husbandry and feeding practices in order to increase the number of ASF-free farms.

Development of transgenic potato with improved anthocyanin contents using sweet potato IbMYB1 gene (고구마의 IbMYB1 유전자를 이용한 안토시아닌 고함유 형질전환 감자의 개발)

  • Kim, Yun-Hee;Han, Eun-Hee;Kwak, Sang-Soo;Lee, Shin-Woo
    • Journal of Plant Biotechnology
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    • v.45 no.4
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    • pp.364-368
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    • 2018
  • The R2R3-type protein IbMYB1 transcription factor is a key regulator for anthocyanin biosynthesis in the storage roots of sweet potatoes. It was previously demonstrated that the IbMYB1 expression stimulates anthocyanin pigmentation in tobacco leaves, arabidopsis and storage roots of sweet potatoes. In this study, we generated the transgenic potato plants that express the IbMYB1 genes, which accumulated high levels of anthocyanins under the control of either the tuber-specific patatin (PAT) promoter or oxidative stress-inducible peroxidase anionic 2 (SWPA2) promoter. The PAT-MYB1 transgenic lines exhibited higher anthocyanin levels in the tuber than the empty vector control (EV) or SWPA2-MYB1 plants. When combined, our results indicated that overexpression of the IbMYB1 is a highly promising strategy for the generation of transgenic plants with enhanced tissue specific anthocyanin production.

An alpha-lipoic acid-decursinol hybrid compound attenuates lipopolysaccharide-mediated inflammation in BV2 and RAW264.7 cells

  • Kwon, Mi-Youn;Park, Jiwon;Kim, Sang-Min;Lee, Jooweon;Cho, Hyeongjin;Park, Jeong-Ho;Han, Inn-Oc
    • BMB Reports
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    • v.52 no.8
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    • pp.508-513
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    • 2019
  • In this study, the anti-inflammatory effects of ${\alpha}-lipoic$ acid (LA) and decursinol (Dec) hybrid compound LA-Dec were evaluated and compared with its prodrugs, LA and Dec. LA-Dec dose-dependently inhibited lipopolysaccharide (LPS)-induced nitric oxide (NO) generation in BV2 mouse microglial cells. On the other hand, no or mild inhibitory effect was shown by the Dec and LA, respectively. LA-Dec demonstrated dose-dependent protection from activation-induced cell death in BV2 cells. LA-Dec, but not LA or Dec individually, inhibited LPS-induced increased expressions of induced NO synthase (iNOS) and cyclooxygenase-2 (COX-2) proteins in a dose-dependent manner in both BV2 and mouse macrophage, RAW264.7 cells. Furthermore, LA-Dec inhibited LPS-induced expressions of iNOS, COX-2, interleukin-6, tumor necrosis $factor-{\alpha}$, and $interleukin-1{\beta}$ mRNA in BV2 cells, whereas the same concentration of LA or Dec was ineffective. Signaling studies demonstrated that LA-Dec inhibited LPS-activated signal transducer and activator of transcription 3 and protein kinase B activation, but not nuclear factor-kappa B or mitogen-activated protein kinase signaling. The data implicate LA-Dec hybrid compound as a potential therapeutic agent for inflammatory diseases of the peripheral and central nervous systems.

Spatial and temporal dynamic of land-cover/land-use and carbon stocks in Eastern Cameroon: a case study of the teaching and research forest of the University of Dschang

  • Temgoua, Lucie Felicite;Solefack, Marie Caroline Momo;Voufo, Vianny Nguimdo;Belibi, Chretien Tagne;Tanougong, Armand
    • Forest Science and Technology
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    • v.14 no.4
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    • pp.181-191
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    • 2018
  • This study was carried out in the teaching and research forest of the University of Dschang in Belabo, with the aim of analysing land-cover and land-use changes as well as carbon stocks dynamic. The databases used are composed of three Landsat satellite images (5TM of 1984, 7ETM + of 2000 and 8OLI of 2016), enhanced by field missions. Satellite images were processed using ENVI and ArcGIS software. Interview, focus group discussion methods and participatory mapping were used to identify the activities carried out by the local population. An inventory design consisting of four transects was used to measure dendrometric parameters and to identify land-use types. An estimation of carbon stocks in aboveground and underground woody biomass was made using allometric models based on non-destructive method. Dynamic of land-cover showed that the average annual rate of deforestation is 0.48%. The main activities at the base of this change are agriculture, house built-up and logging. Seven types of land-use were identified; adult secondary forests (64.10%), young secondary forests (7.54%), wetlands (7.39%), fallows (3.63%), savannahs (9.59%), cocoa farms (4.28%) and mixed crop farms (3.47%). Adult secondary forests had the highest amount of carbon ($250.75\;t\;C\;ha^{-1}$). This value has decreased by more than 60% for mixed crop farms ($94.67\;t\;C\;ha^{-1}$), showing the impact of agricultural activities on both forest cover and carbon stocks. Agroforestry systems that allow conservation and introduction of woody species should be encouraged as part of a participatory management strategy of this forest.

Albendazole and Mebendazole as Anti-Parasitic and Anti-Cancer Agents: an Update

  • Chai, Jong-Yil;Jung, Bong-Kwang;Hong, Sung-Jong
    • Parasites, Hosts and Diseases
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    • v.59 no.3
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    • pp.189-225
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    • 2021
  • The use of albendazole and mebendazole, i.e., benzimidazole broad-spectrum anthelmintics, in treatment of parasitic infections, as well as cancers, is briefly reviewed. These drugs are known to block the microtubule systems of parasites and mammalian cells leading to inhibition of glucose uptake and transport and finally cell death. Eventually they exhibit ovicidal, larvicidal, and vermicidal effects on parasites, and tumoricidal effects on hosts. Albendazole and mebendazole are most frequently prescribed for treatment of intestinal nematode infections (ascariasis, hookworm infections, trichuriasis, strongyloidiasis, and enterobiasis) and can also be used for intestinal tapeworm infections (taeniases and hymenolepiasis). However, these drugs also exhibit considerable therapeutic effects against tissue nematode/cestode infections (visceral, ocular, neural, and cutaneous larva migrans, anisakiasis, trichinosis, hepatic and intestinal capillariasis, angiostrongyliasis, gnathostomiasis, gongylonemiasis, thelaziasis, dracunculiasis, cerebral and subcutaneous cysticercosis, and echinococcosis). Albendazole is also used for treatment of filarial infections (lymphatic filariasis, onchocerciasis, loiasis, mansonellosis, and dirofilariasis) alone or in combination with other drugs, such as ivermectin or diethylcarbamazine. Albendazole was tried even for treatment of trematode (fascioliasis, clonorchiasis, opisthorchiasis, and intestinal fluke infections) and protozoan infections (giardiasis, vaginal trichomoniasis, cryptosporidiosis, and microsporidiosis). These drugs are generally safe with few side effects; however, when they are used for prolonged time (>14-28 days) or even only 1 time, liver toxicity and other side reactions may occur. In hookworms, Trichuris trichiura, possibly Ascaris lumbricoides, Wuchereria bancrofti, and Giardia sp., there are emerging issues of drug resistance. It is of particular note that albendazole and mebendazole have been repositioned as promising anti-cancer drugs. These drugs have been shown to be active in vitro and in vivo (animals) against liver, lung, ovary, prostate, colorectal, breast, head and neck cancers, and melanoma. Two clinical reports for albendazole and 2 case reports for mebendazole have revealed promising effects of these drugs in human patients having variable types of cancers. However, because of the toxicity of albendazole, for example, neutropenia due to myelosuppression, if high doses are used for a prolonged time, mebendazole is currently more popularly used than albendazole in anti-cancer clinical trials.

Ethanol inhibits Kv7.2/7.3 channel open probability by reducing the PI(4,5)P2 sensitivity of Kv7.2 subunit

  • Kim, Kwon-Woo;Suh, Byung-Chang
    • BMB Reports
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    • v.54 no.6
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    • pp.311-316
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    • 2021
  • Ethanol often causes critical health problems by altering the neuronal activities of the central and peripheral nerve systems. One of the cellular targets of ethanol is the plasma membrane proteins including ion channels and receptors. Recently, we reported that ethanol elevates membrane excitability in sympathetic neurons by inhibiting Kv7.2/7.3 channels in a cell type-specific manner. Even though our studies revealed that the inhibitory effects of ethanol on the Kv7.2/7.3 channel was diminished by the increase of plasma membrane phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2), the molecular mechanism of ethanol on Kv7.2/7.3 channel inhibition remains unclear. By investigating the kinetics of Kv7.2/7.3 current in high K+ solution, we found that ethanol inhibited Kv7.2/7.3 channels through a mechanism distinct from that of tetraethylammonium (TEA) which enters into the pore and blocks the gate of the channels. Using a non-stationary noise analysis (NSNA), we demonstrated that the inhibitory effect of ethanol is the result of reduction of open probability (PO) of the Kv7.2/7.3 channel, but not of a single channel current (i) or channel number (N). Finally, ethanol selectively facilitated the kinetics of Kv7.2 current suppression by voltage-sensing phosphatase (VSP)-induced PI(4,5)P2 depletion, while it slowed down Kv7.2 current recovery from the VSP-induced inhibition. Together our results suggest that ethanol regulates neuronal activity through the reduction of open probability and PI(4,5)P2 sensitivity of Kv7.2/7.3 channels.

Analysis of Tissue-Specific Interferon Regulatory Factor 3 (IRF3) Gene Expression against Viral Infection in Paralichthys olivaceus

  • Kim, Kyung-Hee;Lee, Sanghyun;Park, Jong-Won;Jung, Hyo Sun;Kim, Julan;Yang, Hyerim;Lee, Jeong-Ho;Lee, Dain
    • Development and Reproduction
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    • v.25 no.4
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    • pp.235-244
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    • 2021
  • Interferon Regulatory Factor 3 (IRF3) is a member of interferon-regulated transcription factor family and is known to play an important role in the innate immune response against viral infections. In this study, the expression of IRF3 in different tissues, developmental stages, and stocking densities of olive flounder was investigated. The expression of IRF3 was observed to gradually increase in early-stage juvenile fish. The highest expression was observed in later-stage juvenile fish when immune tissues were formed. High IRF3 expression was observed in the muscles and the brain tissues. The expression of IRF3 was observed in fish at different stocking densities after viral hemorrhagic septicemia virus (VHSV) infection. It yielded an interesting expression pattern in the muscles and the brain tissues of fish stocked at low density. These observations can be used as basic data for the study of the expression of immune response-related genes against viruses based on stocking density and immune systems in other fish species.

Pathogenesis and prognosis of primary oral squamous cell carcinoma based on microRNAs target genes: a systems biology approach

  • Taherkhani, Amir;Dehto, Shahab Shahmoradi;Jamshidi, Shokoofeh;Shojaei, Setareh
    • Genomics & Informatics
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    • v.20 no.3
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    • pp.27.1-27.13
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    • 2022
  • Oral squamous cell carcinoma (OSCC) is the most prevalent head and neck malignancy, with frequent cervical lymph-node metastasis, leading to a poor prognosis in OSCC patients. The present study aimed to identify potential markers, including microRNAs (miRNAs) and genes, significantly involved in the etiology of early-stage OSCC. Additionally, the main OSCC's dysregulated Gene Ontology annotations and significant signaling pathways were identified. The dataset GSE45238 underwent multivariate statistical analysis in order to distinguish primary OSCC tissues from healthy oral epithelium. Differentially expressed miRNAs (DEMs) with the criteria of p-value < 0.001 and |Log2 fold change| > 1.585 were identified in the two groups, and subsequently, validated targets of DEMs were identified. A protein interaction map was constructed, hub genes were identified, significant modules within the network were illustrated, and significant pathways and biological processes associated with the clusters were demonstrated. Using the GEPI2 database, the hub genes' predictive function was assessed. Compared to the healthy controls, main OSCC had a total of 23 DEMs. In patients with head and neck squamous cell carcinoma (HNSCC), upregulation of CALM1, CYCS, THBS1, MYC, GATA6, and SPRED3 was strongly associated with a poor prognosis. In HNSCC patients, overexpression of PIK3R3, GIGYF1, and BCL2L11 was substantially correlated with a good prognosis. Besides, "proteoglycans in cancer" was the most significant pathway enriched in the primary OSCC. The present study results revealed more possible mechanisms mediating primary OSCC and may be useful in the prognosis of the patients with early-stage OSCC.

A report of 43 unrecorded bacterial species within the phyla Bacteroidetes and Firmicutes isolated from various sources from Korea in 2019

  • Kang, Heeyoung;Kim, Haneul;Yi, Hana;Kim, Wonyong;Yoon, Jung-Hoon;Im, Wan-Taek;Kim, Myung Kyum;Seong, Chi Nam;Kim, Seung Bum;Cha, Chang-Jun;Jeon, Che Ok;Joh, Kiseong
    • Journal of Species Research
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    • v.10 no.2
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    • pp.117-133
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    • 2021
  • In 2019, 43 bacterial strains were isolated from food, soil, marine environments, human, and animals related sources from the Republic of Korea. Based on the analysis of 16S rRNA gene sequence, these isolates were allocated to the phyla Bacteroidetes and Firmicutes as unrecorded species in Korea. The 10 Bacteroidetes strains were classified into the families Bacteroidaceae, Chitinophagaceae, Cytophagaceae, Flavobacteriaceae, and Prolixibacteraceae (of the orders Bacteroidales, Chitinophagales, Cytophagales, Flavobacteriales, and Marinilabiliales, respectively). The 33 Firmicutes strains belonged to the families Bacillaceae, Paenibacillaceae, Planococcaceae, Staphylococcaceae, Clostridiaceae, Lachnospiraceae, Peptostreptococcaceae, Enterococcaceae, Lactobacillaceae, Leuconostocaceae, and Streptococcaceae (of the orders Bacillales, Clostridiales, and Lactobacillales). These unrecorded bacteria were determined based on taxonomic criterion (>98.7%; 16S rRNA gene sequence similarity). In addition, their phylogenetic affiliation, as well as cell and colony morphologies, staining reactions, and physiological and biochemical properties were investigated. Therefore, we report 43 isolates as unrecorded species, and described basic features, isolation source, and locations of these strains.