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1.
Mol Cell Proteomics ; 22(10): 100637, 2023 Oct.
Article in English | MEDLINE | ID: mdl-37648026

ABSTRACT

cAMP response element-binding protein (CREB) regulated transcriptional coactivator 2 (CRTC2) is a critical transcription factor that maintains glucose homeostasis by activating CREB. Energy homeostasis is maintained through multiple pathways; therefore, CRTC2 may interact with other transcription factors, particularly under metabolic stress. CRTC2 liver-specific KO mice were created, and the global proteome, phosphoproteome, and acetylome from liver tissue under high-fat diet conditions were analyzed using liquid chromatography-tandem mass spectrometry and bioinformatics analysis. Differentially regulated proteins (DRPs) were enriched in metabolic pathways, which were subsequently corroborated through animal experiments. The consensus DRPs from these datasets were used as seed proteins to generate a protein-protein interaction network using STRING, and GeneMANIA identified fatty acid synthase as a mutually relevant protein. In an additional local-protein-protein interaction analysis of CRTC2 and fatty acid synthase with DRPs, sterol regulatory element binding transcription factor 1 (SREBF1) was the common mediator. CRTC2-CREB and SREBF1 are transcription factors, and DNA-binding motif analysis showed that multiple CRTC2-CREB-regulated genes possess SREBF1-binding motifs. This indicates the possible induction by the CRTC2-SREBF1 complex, which is validated through luciferase assay. Therefore, the CRTC2-SREBF1 complex potentially modulates the transcription of multiple proteins that fine-tune cellular metabolism under metabolic stress.

2.
Proteomics ; 24(11): e2300062, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38829178

ABSTRACT

Extracellular vesicles (EVs) are membrane-surrounded vesicles released by various cell types into the extracellular microenvironment. Although EVs vary in size, biological function, and components, their importance in cancer progression and the potential use of EV molecular species to serve as novel cancer biomarkers have become increasingly evident. Cancer cells actively release EVs into surrounding tissues, which play vital roles in cancer progression and metastasis, including invasion and immune modulation. EVs released by cancer cells are usually chosen as a gateway in the search for biomarkers for cancer. In this review, we mainly focused on molecular profiling of EV protein constituents from breast cancer, emphasizing mass spectrometry (MS)-based proteomic approaches. To further investigate the potential use of EVs as a source of breast cancer biomarkers, we have discussed the use of these proteins as predictive marker candidates. Besides, we have also summarized the key characteristics of EVs as potential therapeutic targets in breast cancer and provided significant information on their implications in breast cancer development and progression. Information provided in this review may help understand the recent progress in understanding EV biology and their potential role as new noninvasive biomarkers as well as emerging therapeutic opportunities and associated challenges.


Subject(s)
Biomarkers, Tumor , Breast Neoplasms , Extracellular Vesicles , Mass Spectrometry , Proteomics , Humans , Breast Neoplasms/metabolism , Breast Neoplasms/pathology , Biomarkers, Tumor/metabolism , Biomarkers, Tumor/analysis , Extracellular Vesicles/metabolism , Female , Mass Spectrometry/methods , Proteomics/methods
3.
EMBO J ; 39(6): e102214, 2020 03 16.
Article in English | MEDLINE | ID: mdl-32030804

ABSTRACT

Spinal cord microglia contribute to nerve injury-induced neuropathic pain. We have previously demonstrated that toll-like receptor 2 (TLR2) signaling is critical for nerve injury-induced activation of spinal cord microglia, but the responsible endogenous TLR2 agonist has not been identified. Here, we show that nerve injury-induced upregulation of sialyltransferase St3gal2 in sensory neurons leads to an increase in expression of the sialylated glycosphingolipid, GT1b. GT1b ganglioside is axonally transported to the spinal cord dorsal horn and contributes to characteristics of neuropathic pain such as mechanical and thermal hypersensitivity. Spinal cord GT1b functions as an TLR2 agonist and induces proinflammatory microglia activation and central sensitization. Pharmacological inhibition of GT1b synthesis attenuates nerve injury-induced spinal cord microglia activation and pain hypersensitivity. Thus, the St3gal2-GT1b-TLR2 axis may offer a novel therapeutic target for the treatment of neuropathic pain.


Subject(s)
Gangliosides/metabolism , Neuralgia/therapy , Peripheral Nerve Injuries/therapy , Signal Transduction , Toll-Like Receptor 2/agonists , Animals , Gangliosides/antagonists & inhibitors , Gene Expression Regulation , Inflammation , Male , Mice , Mice, Inbred C57BL , Microglia/metabolism , Neuralgia/etiology , Peripheral Nerve Injuries/etiology , Rats , Rats, Sprague-Dawley , Sensory Receptor Cells , Sialyltransferases/genetics , Sialyltransferases/metabolism , Spinal Cord/metabolism , Toll-Like Receptor 2/metabolism
4.
Biochem Biophys Res Commun ; 720: 150098, 2024 Aug 06.
Article in English | MEDLINE | ID: mdl-38749190

ABSTRACT

Telomerase reverse transcriptase (TERT) not only upholds telomeric equilibrium but also plays a pivotal role in multiple non-canonical cellular mechanisms, particularly in the context of aging, cancer, and genomic stability. Though depletion of SIRT1 in mouse embryonic fibroblasts has demonstrated telomere shortening, the impact of SIRT1 on enabling TERT to regulate telomeric homeostasis remains enigmatic. Here, we reveal that SIRT1 directly interacts with TERT, and promotes the nuclear localization and stability of TERT. Reverse transcriptase (RT) domain of TERT and N-terminus of SIRT1 mainly participated in their direct interaction. TERT, concomitantly expressed with intact SIRT1, exhibits nuclear localization, whereas TERT co-expressed with N-terminal-deleted SIRT1 remains in the cytosol. Furthermore, overexpression of SIRT1 enhances the nuclear localization and protein stability of TERT, akin to overexpression of deacetylase-inactive SIRT1, whereas N-terminal-deleted SIRT1 has no effect on TERT. These findings suggest a novel regulatory role of SIRT1 for TERT through direct interaction. This interaction provides new insights into the fields of aging, cancer, and genome stability governed by TERT and SIRT1.


Subject(s)
Sirtuin 1 , Telomerase , Animals , Humans , Mice , Cell Nucleus/metabolism , Enzyme Stability , HEK293 Cells , Protein Binding , Protein Stability , Sirtuin 1/metabolism , Sirtuin 1/genetics , Telomerase/metabolism , Telomerase/genetics
5.
Mol Psychiatry ; 28(2): 810-821, 2023 02.
Article in English | MEDLINE | ID: mdl-36253443

ABSTRACT

Autism spectrum disorder (ASD) is a major neurodevelopmental disorder in which patients present with core symptoms of social communication impairment, restricted interest, and repetitive behaviors. Although various studies have been performed to identify ASD-related mechanisms, ASD pathology is still poorly understood. CNTNAP2 genetic variants have been found that represent ASD genetic risk factors, and disruption of Cntnap2 expression has been associated with ASD phenotypes in mice. In this study, we performed an integrative multi-omics analysis by combining quantitative proteometabolomic data obtained with Cntnap2 knockout (KO) mice with multi-omics data obtained from ASD patients and forebrain organoids to elucidate Cntnap2-dependent molecular networks in ASD. To this end, a mass spectrometry-based proteometabolomic analysis of the medial prefrontal cortex in Cntnap2 KO mice led to the identification of Cntnap2-associated molecular features, and these features were assessed in combination with multi-omics data obtained on the prefrontal cortex in ASD patients to identify bona fide ASD cellular processes. Furthermore, a reanalysis of single-cell RNA sequencing data obtained from forebrain organoids derived from patients with CNTNAP2-associated ASD revealed that the aforementioned identified ASD processes were mainly linked to excitatory neurons. On the basis of these data, we constructed Cntnap2-associated ASD network models showing mitochondrial dysfunction, axonal impairment, and synaptic activity. Our results may shed light on the Cntnap2-dependent molecular networks in ASD.


Subject(s)
Autism Spectrum Disorder , Mice , Animals , Multiomics , Mice, Knockout , Neurons/metabolism , Axons/metabolism , Membrane Proteins/genetics , Membrane Proteins/metabolism , Nerve Tissue Proteins/metabolism
6.
Eur Radiol ; 2024 Feb 15.
Article in English | MEDLINE | ID: mdl-38358528

ABSTRACT

OBJECTIVES: The carcinogenic risks of CT radiation in children and adolescents remain debated. We aimed to assess the carcinogenic risk of CTs performed in children and adolescents with minor head trauma. METHODS: In this nationwide population-based cohort study, we included 2,411,715 patients of age 0-19 with minor head trauma from 2009 to 2017. We excluded patients with elevated cancer risks or substantial past medical radiation exposure. Patients were categorized into CT-exposed or CT-unexposed group according to claim codes for head CT. The primary outcome was development of hematologic malignant neoplasms. Secondary outcomes included development of malignant solid neoplasms and benign neoplasms in the brain. We measured the incidence rate ratio (IRR) and incidence rate difference (IRD) using G-computation with Poisson regression adjusting for age, sex, hospital setting, and the type of head trauma. RESULTS: Hematologic malignant neoplasms developed in 100 of 216,826 patients during 1,303,680 person-years in the CT-exposed group and in 808 of 2,194,889 patients during 13,501,227 person-years in the CT-unexposed group. For hematologic malignant neoplasms, the IRR was 1.29 (95% CI, 1.03-1.60) and the IRD was 1.71 (95% CI, 0.04-3.37) per 100,000 person-years at risk. The majority of excess hematologic malignant neoplasms were leukemia (IRR, 1.40 [98.3% CI, 1.05-1.87]; IRD, 1.59 [98.3% CI, 0.02-3.16] per 100,000 person-years at risk). There were no between-group differences for secondary outcomes. CONCLUSIONS: Radiation exposure from head CTs in children and adolescents with minor head trauma was associated with an increased incidence of hematologic malignant neoplasms. CLINICAL RELEVANCE STATEMENT: Our study provides a quantitative grasp of the risk conferred by CT examinations in children and adolescents, thereby providing the basis for cost-benefit analyses and evidence-driven guidelines for patient triaging in head trauma. KEY POINTS: • This nationwide population-based cohort study showed that radiation exposure from head CTs in children and adolescents was associated with a higher incidence of hematologic malignant neoplasms. • The incidence rate of hematologic malignant neoplasms in the CT-exposed group was 29% higher than that in the CT-unexposed group (IRR, 1.29 [95% CI, 1.03-1.60]), and there were approximately 1.7 excess neoplasms per 100,000 person-years at risk in the CT-exposed group (IRD, 1.71 [0.04-3.37]). • Our study provides a quantified grasp of the risk conferred by CT examinations in children and adolescents, while controlling for biases observed in previous studies via specifying CT indication and excluding patients with predisposing conditions for cancer development.

7.
RNA Biol ; 21(1): 1-18, 2024 Jan.
Article in English | MEDLINE | ID: mdl-38361426

ABSTRACT

Protein aggregation, a consequence of misfolding and impaired proteostasis, can lead to cellular malfunctions such as various proteinopathies. The mechanisms protecting proteins from aggregation in complex cellular environments have long been investigated, often from a protein-centric viewpoint. However, our study provides insights into a crucial, yet overlooked actor: RNA. We found that depleting RNAs from Escherichia coli lysates induces global protein aggregation. Our quantitative mass spectrometry analysis identified over 900 statistically significant proteins from the Escherichia coli proteome whose solubility depends on RNAs. Proteome-wide characterization showed that the RNA dependency is particularly enriched among acidic proteins, intrinsically disordered proteins, and structural hub proteins. Moreover, we observed distinct differences in RNA-binding mode and Gene Ontology categories between RNA-dependent acidic and basic proteins. Notably, the solubility of key molecular chaperones [Trigger factor, DnaJ, and GroES] is largely dependent on RNAs, suggesting a yet-to-be-explored hierarchical relationship between RNA-based chaperone (termed as chaperna) and protein-based chaperones, both of which constitute the whole chaperone network. These findings provide new insights into the RNA-centric role in maintaining healthy proteome solubility in vivo, where proteins associate with a variety of RNAs, either stably or transiently.


Subject(s)
Escherichia coli Proteins , Escherichia coli , Escherichia coli/genetics , Escherichia coli/metabolism , Proteome/metabolism , Protein Folding , RNA/metabolism , Solubility , Proteomics , Isoelectric Point , Protein Aggregates , Escherichia coli Proteins/metabolism , Molecular Chaperones/genetics , Molecular Chaperones/metabolism , Mass Spectrometry
8.
J Radiol Prot ; 44(1)2024 Mar 28.
Article in English | MEDLINE | ID: mdl-38507784

ABSTRACT

In Korea, decommissioning of nuclear power plants and transportation of the decommissioning waste are expected to expand in the near future. It is necessary to confirm that radiological risks to the public and workers are not significant through radiological safety assessment. The objective of this study is to assess the radiological safety for transportation of RPV waste, which is a major decommissioning waste with relatively high level of radioactivity. It was assumed that the waste would be transported to the Gyeongju disposal facility by land transportation. First, the source term and transportation method of the RPV waste were determined, and the external dose rates from the waste were calculated using MCNP. Then, transportation scenarios were assumed under both normal and accident conditions. Under the scenarios, radiation doses were calculated using the RADTRAN. Under normal operation scenarios without a transportation accident, assuming 40 shipments per year, the average individual doses for the public ranged from 6.56×10-6to 2.18×10-2mSv yr-1. The maximum individual doses for only a single shipment ranged from 2.43×10-6to 3.14×10-1mSv. For cargo handlers and vehicle crew members, the average doses were 2.26×101mSv yr-1and 2.95 mSv yr-1, respectively. Under transportation accident scenarios, average individual radiological risks which are product of the radiation doses and the annual accident rates ranged from 1.14×10-11to 1.61×10-10mSv yr-1by transportation route segment when considering the transportation accident rate. Average individual doses assuming transportation accident occurrence ranged from 2.62×10-4to 1.42×10-3mSv. The maximum individual dose under accident conditions was 7.99×10-2mSv. The calculated doses were below the regulatory limits in Korea. However, relatively high doses were observed for cargo handlers and vehicle crew members because of conservative assumptions. This study results can be used as basic data for the radiological safety assessment for the decommissioning waste transportation in the future.


Subject(s)
Fukushima Nuclear Accident , Radiation Monitoring , Humans , Nuclear Power Plants , Radiation Dosage , Radiation Monitoring/methods , Republic of Korea
9.
EMBO J ; 38(10)2019 05 15.
Article in English | MEDLINE | ID: mdl-30979777

ABSTRACT

The multifunctional influenza virus protein PB1-F2 plays several roles in deregulation of host innate immune responses and is a known immunopathology enhancer of the 1918 influenza pandemic. Here, we show that the 1918 PB1-F2 protein not only interferes with the mitochondria-dependent pathway of type I interferon (IFN) signaling, but also acquired a novel IFN antagonist function by targeting the DEAD-box helicase DDX3, a key downstream mediator in antiviral interferon signaling, toward proteasome-dependent degradation. Interactome analysis revealed that 1918 PB1-F2, but not PR8 PB1-F2, binds to DDX3 and causes its co-degradation. Consistent with intrinsic protein instability as basis for this gain-of-function, internal structural disorder is associated with the unique cytotoxic sequences of the 1918 PB1-F2 protein. Infusing mice with recombinant DDX3 protein completely rescued them from lethal infection with the 1918 PB1-F2-producing virus. Alongside NS1 protein, 1918 PB1-F2 therefore constitutes a potent IFN antagonist causative for the severe pathogenicity of the 1918 influenza strain. Our identification of molecular determinants of pathogenesis should be useful for the future design of new antiviral strategies against influenza pandemics.


Subject(s)
DEAD-box RNA Helicases/metabolism , Influenza, Human/virology , Interferons/metabolism , Orthomyxoviridae/pathogenicity , Viral Proteins/physiology , A549 Cells , Animals , Dogs , Female , HEK293 Cells , History, 20th Century , Humans , Influenza, Human/epidemiology , Influenza, Human/history , Madin Darby Canine Kidney Cells , Mice , Mice, Inbred BALB C , Orthomyxoviridae/metabolism , Pandemics , Proteolysis , Signal Transduction , U937 Cells , Viral Proteins/metabolism , Virulence/physiology
10.
Pharmacol Res ; 187: 106610, 2023 Jan.
Article in English | MEDLINE | ID: mdl-36521573

ABSTRACT

Gastric cancer (GC) occurs in the gastric mucosa, and its high morbidity and mortality make it an international health crisis. Therefore, novel drugs are needed for its treatment. The use of natural products and their components in cancer treatments has shown promise. Therefore, this study aimed to evaluate the effect of 8-paradol, a phenolic compound isolated from ginger (Zingiber officinale Roscoe), on GC and determine its underlying mechanisms of action. In this study, repeated column chromatography was conducted on ginger EtOH extract to isolate gingerol and its derivatives. The cytotoxicity of the eight ginger compounds underwent a (3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide) tetrazolium reduction (MTT) assay. 8-paradol showed the most potent cytotoxicity effect among the isolated ginger compounds. The underlying mechanism by which 8-paradol regulated specific proteins in AGS cells was evaluated by proteomic analysis. To validate the predicted mechanisms, AGS cells and thymus-deficient nude mice bearing AGS xenografts were used as in vitro and in vivo models of GC, respectively. The results showed that the 8-paradol promoted PINK1/Parkin-associated mitophagy, mediating cell apoptosis. Additionally, the inhibition of mitophagy by chloroquine (CQ) ameliorated 8-paradol-induced mitochondrial dysfunction and apoptosis, supporting a causative role for mitophagy in the 8-paradol-induced anticancer effect. Molecular docking results revealed the molecular interactions between 8-paradol and mitophagy-/ apoptosis-related proteins at the atomic level. Our study provides strong evidence that 8-paradol could act as a novel potential therapeutic agent to suppress the progression of GC by targeting mitophagy pathway.


Subject(s)
Adenocarcinoma , Stomach Neoplasms , Zingiber officinale , Mice , Animals , Humans , Zingiber officinale/chemistry , Zingiber officinale/metabolism , Mitophagy , Stomach Neoplasms/drug therapy , Mice, Nude , Molecular Docking Simulation , Proteomics , Apoptosis , Protein Kinases/metabolism , Ubiquitin-Protein Ligases/metabolism
11.
Virus Genes ; 59(4): 624-634, 2023 Aug.
Article in English | MEDLINE | ID: mdl-37119398

ABSTRACT

In the present study, two new Bacillus subtilis phages, BSTP4 and BSTP6, were isolated and studied further. Morphologically, BSTP4 and BSTP6 are podoviruses with complete genome of 19,145 (39.9% G + C content) and 19,367 bp (39.8% G + C content), respectively, which became among the smallest Bacillus phages. Three most prominent structural proteins were separated and identified as pre-neck appendage, major head, and head fiber proteins using LC-MS/MS. Both phages encode putative terminal proteins (TP) and contain short inverted terminal repeats (ITRs) which may be important for their replication. In addition, non-coding RNA (pRNA) and parS sites were identified which may be required for DNA packaging and their maintenance inside the host, respectively. Furthermore, the phage genome sequences show significant similarity to B. subtilis group species genome sequences. Finally, phylogenomic and phylogenetic analyses suggest that BSTP4 and BSTP6 may form a new species in the genus Salasvirus, subfamily Picovirinae of family Salasmaviridae. Considering the small numbers of ICTV-accepted B. subtilis phages and the importance of the host in the food industry and biotechnology, the current study helps to improve our understanding of the diversity of B. subtilis phages and shed light on the phage-host relationships.


Subject(s)
Bacillus Phages , Podoviridae , Bacillus subtilis/genetics , Phylogeny , Chromatography, Liquid , Genome, Viral , Tandem Mass Spectrometry , Podoviridae/genetics , Bacillus Phages/genetics , Sequence Analysis
12.
Mol Cell Proteomics ; 20: 100017, 2021.
Article in English | MEDLINE | ID: mdl-33592500

ABSTRACT

Extracellular vesicle (EV) proteins from acute myeloid leukemia (AML) cell lines were analyzed using mass spectrometry. The analyses identified 2450 proteins, including 461 differentially expressed proteins (290 upregulated and 171 downregulated). CD53 and CD47 were upregulated and were selected as candidate biomarkers. The association between survival of patients with AML and the expression levels of CD53 and CD47 at diagnosis was analyzed using mRNA expression data from The Cancer Genome Atlas database. Patients with higher expression levels showed significantly inferior survival than those with lower expression levels. ELISA results of the expression levels of CD53 and CD47 from EVs in the bone marrow of patients with AML at diagnosis and at the time of complete remission with induction chemotherapy revealed that patients with downregulated CD53 and CD47 expression appeared to relapse less frequently. Network model analysis of EV proteins revealed several upregulated kinases, including LYN, CSNK2A1, SYK, CSK, and PTK2B. The potential cytotoxicity of several clinically applicable drugs that inhibit these kinases was tested in AML cell lines. The drugs lowered the viability of AML cells. The collective data suggest that AML cell-derived EVs could reflect essential leukemia biology.


Subject(s)
Biomarkers, Tumor/metabolism , Extracellular Vesicles/metabolism , Leukemia, Myeloid, Acute/metabolism , Adolescent , Adult , Aged , Antigens, CD/genetics , Antigens, CD/metabolism , Cells, Cultured , Female , Humans , Male , Middle Aged , Protein Kinases/metabolism , Proteomics , Young Adult
13.
Ecotoxicol Environ Saf ; 262: 115119, 2023 Jun 14.
Article in English | MEDLINE | ID: mdl-37327520

ABSTRACT

Eicosanoids play important roles in inflammation, allergy, fever, and immune responses. In the eicosanoid pathway, cyclooxygenase (COX) catalyzes the conversion of arachidonic acid to prostaglandins and is a crucial target of nonsteroidal anti-inflammatory drugs (NSAIDs). Thus, toxicological studies on the eicosanoid pathway are important for drug discovery and the evaluation of adverse health outcomes due to environmental contaminants. However, experimental models are limited owing to concerns regarding ethical standards. Thus, new alternative models for evaluating toxic effects on the eicosanoid pathway must be developed. To this end, we adopted an invertebrate species, Daphnia magna, as an alternative model. D. magna was exposed to ibuprofen, a major NSAID, for 6 and 24 h. Transcription of eicosanoid-related genes (pla2, cox, pgd synthase, pgd2r2, ltb4dh, and lox) was analyzed by qPCR, eicosanoids (arachidonic acid, prostaglandin F2, dihydroxy prostaglandin F2, and 5-hydroxyeicosatetraenoate) were quantified by multiple reaction monitoring, and enzyme-linked immunosorbent assay was used to determine protein levels of arachidonic acid and prostaglandin E2 (PGE2). After 6 h of exposure, transcription of the pla2 and cox genes was downregulated. In addition, the whole-body level of arachidonic acid, an upstream of COX pathway, increased by over 1.5-fold. The levels of PGE2, a downstream of COX pathway, decreased after 24 h of exposure. According to our results, it is expected that the eicosanoid pathway might be conserved in D. magna, at least partially. This indicates the plausibility of D. magna as an alternative model for the screening of new drugs or chemical toxicity.

14.
J Radiol Prot ; 43(2)2023 04 24.
Article in English | MEDLINE | ID: mdl-36996806

ABSTRACT

The objective of this study is to update the national and regional indoor radon concentrations in South Korea and assess indoor radon exposure. Based on the previously published survey results and the collected measurement data of surveys conducted since 2011, a total of 9271 indoor radon measurement data covering 17 administrative divisions are finally used for analysis. The annual effective dose from the indoor radon exposure is calculated using dose coefficients recommended by the International Commission on Radiological Protection. The population-weighted average indoor radon concentration was estimated to be a geometric mean of 46 Bq m-3(GSD = 1.2) with 3.9% of all samples showing values exceeding 300 Bq m-3. The regional average indoor radon concentration ranged from 34 to 73 Bq m-3. The radon concentrations in detached houses were relatively higher than those in public buildings and multi-family houses. The annual effective doses to the Korean population due to indoor radon exposure were estimated to be 2.18 mSv. The updated values in this study might better represent the national indoor radon exposure level in South Korea because they contain more samples and cover a wider range of geographical areas than previous studies.


Subject(s)
Air Pollutants, Radioactive , Air Pollution, Indoor , Radiation Monitoring , Radon , Air Pollutants, Radioactive/analysis , Air Pollution, Indoor/analysis , Radon/analysis , Republic of Korea , Housing
15.
BMC Genomics ; 23(1): 835, 2022 Dec 16.
Article in English | MEDLINE | ID: mdl-36526963

ABSTRACT

BACKGROUND: Despite the applications of Bacillus subtilis group species in various sectors, limited information is available regarding their phages. Here, 61 B. subtilis group species-infecting phages (BSPs) were studied for their taxonomic classification considering the genome-size, genomic diversity, and the host, followed by the identification of orthologs taxonomic signature genes. RESULTS: BSPs have widely ranging genome sizes that can be bunched into groups to demonstrate correlations to family and subfamily classifications. Comparative analysis re-confirmed the existing, BSPs-containing 14 genera and 21 species and displayed inter-genera similarities within existing subfamilies. Importantly, it also revealed the need for the creation of new taxonomic classifications, including 28 species, nine genera, and two subfamilies (New subfamily1 and New subfamily2) to accommodate inter-genera relatedness. Following pangenome analysis, no ortholog shared by all BSPs was identified, while orthologs, namely, the tail fibers/spike proteins and poly-gamma-glutamate hydrolase, that are shared by more than two-thirds of the BSPs were identified. More importantly, major capsid protein (MCP) type I, MCP type II, MCP type III and peptidoglycan binding proteins that are distinctive orthologs for Herelleviridae, Salasmaviridae, New subfamily1, and New subfamily2, respectively, were identified and analyzed which could serve as signatures to distinguish BSP members of the respective taxon. CONCLUSIONS: In this study, we show the genomic diversity and propose a comprehensive classification of 61 BSPs, including the proposition for the creation of two new subfamilies, followed by the identification of orthologs taxonomic signature genes, potentially contributing to phage taxonomy.


Subject(s)
Bacillus , Bacteriophages , Bacteriophages/genetics , Bacillus/genetics , Bacillus subtilis/genetics , Genomics , Genome, Viral , Phylogeny
16.
Int J Mol Sci ; 23(9)2022 May 07.
Article in English | MEDLINE | ID: mdl-35563621

ABSTRACT

Non-alcoholic steatohepatitis (NASH) has pathological characteristics similar to those of alcoholic hepatitis, despite the absence of a drinking history. The greatest threat associated with NASH is its progression to cirrhosis and hepatocellular carcinoma. The pathophysiology of NASH is not fully understood to date. In this study, we investigated the pathophysiology of NASH from the perspective of glycolysis and the Warburg effect, with a particular focus on microRNA regulation in liver-specific macrophages, also known as Kupffer cells. We established NASH rat and mouse models and evaluated various parameters including the liver-to-body weight ratio, blood indexes, and histopathology. A quantitative phosphoproteomic analysis of the NASH rat model livers revealed the activation of glycolysis. Western blotting and immunohistochemistry results indicated that the expression of pyruvate kinase muscle 2 (PKM2), a rate-limiting enzyme of glycolysis, was upregulated in the liver tissues of both NASH models. Moreover, increases in PKM2 and p-PKM2 were observed in the early phase of NASH. These observations were partially induced by the downregulation of microRNA122-5p (miR-122-5p) and occurred particularly in the Kupffer cells. Our results suggest that the activation of glycolysis in Kupffer cells during NASH was partially induced by the upregulation of PKM2 via miR-122-5p suppression.


Subject(s)
Liver Neoplasms , MicroRNAs , Non-alcoholic Fatty Liver Disease , Pyruvate Kinase/metabolism , Animals , Disease Models, Animal , Down-Regulation , Glycolysis , Kupffer Cells/metabolism , Liver Neoplasms/metabolism , Mice , MicroRNAs/genetics , MicroRNAs/metabolism , Muscles/metabolism , Non-alcoholic Fatty Liver Disease/metabolism , Pyruvate Kinase/genetics , Rats
17.
J Am Chem Soc ; 143(15): 5836-5844, 2021 04 21.
Article in English | MEDLINE | ID: mdl-33834782

ABSTRACT

The identification of each cell type is essential for understanding multicellular communities. Antibodies set as biomarkers have been the main toolbox for cell-type recognition, and chemical probes are emerging surrogates. Herein we report the first small-molecule probe, CDgB, to discriminate B lymphocytes from T lymphocytes, which was previously impossible without the help of antibodies. Through the study of the origin of cell specificity, we discovered an unexpected novel mechanism of membrane-oriented live-cell distinction. B cells maintain higher flexibility in their cell membrane than T cells and accumulate the lipid-like probe CDgB more preferably. Because B and T cells share common ancestors, we tracked the cell membrane changes of the progenitor cells and disclosed the dynamic reorganization of the membrane properties over the lymphocyte differentiation progress. This study casts an orthogonal strategy for the small-molecule cell identifier and enriches the toolbox for live-cell distinction from complex cell communities.


Subject(s)
B-Lymphocytes/cytology , Cell Membrane/metabolism , Fluorescent Dyes/chemistry , T-Lymphocytes/cytology , Animals , B-Lymphocytes/chemistry , B-Lymphocytes/immunology , Bone Marrow Cells/cytology , Bone Marrow Cells/metabolism , Cell Differentiation , Cell Membrane/chemistry , Flow Cytometry , Lipidomics , Mice , T-Lymphocytes/chemistry , T-Lymphocytes/immunology
18.
Eur Radiol ; 31(7): 5160-5171, 2021 Jul.
Article in English | MEDLINE | ID: mdl-33439320

ABSTRACT

OBJECTIVES: To compare image quality and radiation dose between dual-energy subtraction (DES)-based bone suppression images (D-BSIs) and software-based bone suppression images (S-BSIs). METHODS: Chest radiographs (CXRs) of forty adult patients were obtained with the two X-ray devices, one with DES and one with bone suppression software. Three image quality metrics (relative mean absolute error (RMAE), peak signal-to-noise ratio (PSNR), and structural similarity index (SSIM)) between original CXR and BSI for each of D-BSI and S-SBI groups were calculated for each bone and soft tissue areas. Two readers rated the visual image quality for original CXR and BSI for each of D-BSI and S-SBI groups. The dose area product (DAP) values were recorded. Paired t test was used to compare the image quality and DAP values between D-BSI and S-BSI groups. RESULTS: In bone areas, S-BSIs had better SSIM values than D-BSI (94.57 vs. 87.77) but worse RMAE and PSNR values (0.50 vs. 0.20; 20.93 vs. 34.37) (all p < 0.001). In soft tissue areas, S-BSIs had better SSIM values than D-BSI (97.56 vs. 91.42) but similar RMAE and PSNR values (0.29 vs. 0.27; 31.35 vs. 29.87) (all p < 0.001). Both readers gave S-BSIs significantly higher image quality scores than D-BSI (p < 0.001). The mean DAP in software-related images (0.98 dGy·cm2) was significantly lower than that in the DES-related images (1.48 dGy·cm2) (p < 0.001). CONCLUSION: Bone suppression software significantly improved the image quality of bone suppression images with a relatively lower radiation dose, compared with dual-energy subtraction technique. KEY POINTS: • Bone suppression software preserves structure similarity of soft tissues better than dual-energy subtraction technique in bone suppression images. • Bone suppression software achieves superior image quality for lung lesions than dual-energy subtraction technique in bone suppression images. • Bone suppression software can decrease the radiation dose over the hardware-based image processing technique.


Subject(s)
Radiography, Dual-Energy Scanned Projection , Radiography, Thoracic , Adult , Humans , Radiation Dosage , Radiographic Image Interpretation, Computer-Assisted , Software , Subtraction Technique
19.
Arch Virol ; 166(6): 1795-1799, 2021 Jun.
Article in English | MEDLINE | ID: mdl-33839920

ABSTRACT

In this study, bacteriophage BSP7, a novel Bacillus subtilis-infecting member of the family Siphoviridae, was isolated from a Korean soybean-based fermented food, Deonjang, using B. subtilis ATCC 21336 as a host. The genome is 55,455 bp long with 39.92% G+C content. A total of 70 ORFs with no tRNA were detected in the genome. A distinct feature of the BSP7 genome among B. subtilis-infecting Siphoviridae family phages is the presence of putative ORFs related to biosynthesis of 7-cyano-7-deazaguanine (PreQ0), a precursor of queuosine and archaeosine biosynthesis. Bioinformatic analysis revealed that the genome of BSP7 does not exhibit any significant similarities to other phages with sequences in the NCBI database. A comparative genomic analysis also confirmed the uniqueness of BSP7 within the family Siphoviridae.


Subject(s)
Bacillus subtilis/virology , Genome, Viral , Guanine/analogs & derivatives , Siphoviridae/genetics , Base Sequence , DNA, Viral/genetics , Gene Expression Regulation, Viral/physiology , Guanine/biosynthesis , Siphoviridae/isolation & purification , Viral Proteins/genetics , Viral Proteins/metabolism
20.
Cryobiology ; 99: 140-148, 2021 04.
Article in English | MEDLINE | ID: mdl-33242477

ABSTRACT

Cryopreserved oocytes are inevitably exposed to cold stress, which negatively affects the cellular aspects of the oocytes. Lipidomic analysis of the oocytes reveals quantitative changes in lipid classes under conditions of cold stress, leading to potential freezing-vulnerability. We had previously shown that specific phospholipids are significantly downregulated in vitrified-warmed mouse oocytes compared to those in fresh oocytes. In this study, we examined whether supplementation of polyethylene glycol 8000 (PEG 8000) during vitrification influences the lipidome of the oocytes. We used liquid chromatography with tandem mass spectrometry (LC-MS/MS) to study the alteration in the lipidome in three groups of mouse oocytes: fresh, vitrified-warmed, and vitrified with PEG 8000-warmed during vitrification. In these groups, we targeted to analyze 21 lipid classes. We profiled 132 lipid species in the oocytes and statistical analyses revealed lipid classes that were up- or downregulated in these groups. Overall, our data revealed that several classes of lipids were affected during vitrification, and that oocytes vitrified with PEG 8000 to some extent alleviated the levels of changes in phospholipid and sphingolipid contents during vitrification. These results suggest that phospholipids and sphingolipids are influenced by PEG 8000 during vitrification and that PEG 8000 can be considered as a potential candidate for preserving membrane integrity during oocyte cryopreservation.


Subject(s)
Lipidomics , Vitrification , Animals , Chromatography, Liquid , Cryopreservation/methods , Dietary Supplements , Mice , Oocytes , Polyethylene Glycols , Tandem Mass Spectrometry
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