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1.
Artigo em Inglês | MEDLINE | ID: mdl-38869487

RESUMO

A Gram-stain-positive, aerobic bacterium, designated as YPD9-1T, was isolated from the gut contents of a spotty belly greenling, Hexagrammos agrammus, collected near Dokdo island, South Korea. The rod-shaped cells were oxidase-positive, and catalase-negative. The major cellular fatty acids were anteiso-C15 : 0, iso-C15 : 0, C16 : 0, iso-C16 : 0 and iso-C17: 0. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and two unidentified lipids. The DNA G+C content was 47.6 mol% and the predominant respiratory quinone was menaquinone MK-7. The 16S rRNA gene sequence of YPD9-1T showed low sequence similarities to species of the genus Paenibacillus, Paenibacillus pocheonensis Gsoil 1138T (97.21 % of sequence similarity), Paenibacillus aestuarii CJ25T (97.12 %) and Paenibacillus allorhizoplanae JJ-42T (96.89 %). The results of phylogenetic analysis based on 16S rRNA gene sequences indicated that YPD9-1T formed a distinct branch among other species of the genus Paenibacillus. The digital DNA-DNA hybridisation, average nucleotide identity, and average amino acid identity values between YPD9-1T and the related species were in the ranges of 15.3-16.2 %, 74.1-78.4 %, and 71.1-71.9 %, respectively, which are below the species cutoff values. On the basis of the results of the polyphasic analysis, we conclude that strain YPD9-1T represents a novel species of the genus Paenibacillus, for which the name Paenibacillus hexagrammi sp. nov. is proposed. The type strain of Paenibacillus hexagrammi is YPD9-1T (=KCTC 43424T =LMG 32988T).


Assuntos
Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano , Ácidos Graxos , Paenibacillus , Filogenia , RNA Ribossômico 16S , Análise de Sequência de DNA , Vitamina K 2 , RNA Ribossômico 16S/genética , DNA Bacteriano/genética , República da Coreia , Ácidos Graxos/análise , Ácidos Graxos/química , Paenibacillus/isolamento & purificação , Paenibacillus/classificação , Paenibacillus/genética , Vitamina K 2/análogos & derivados , Vitamina K 2/análise , Animais , Hibridização de Ácido Nucleico , Fosfolipídeos/análise , Fosfolipídeos/química
2.
Fish Shellfish Immunol ; 151: 109681, 2024 Jun 12.
Artigo em Inglês | MEDLINE | ID: mdl-38871142

RESUMO

The EGF-containing fibulin-like extracellular matrix protein 2 (EFEMP2) is involved in connective tissue development, elastic fiber formation, and tumor growth. In this study, we characterized the cDNA of EFEMP2 (PoEFEMP2), a member of the fibulin family of ECM proteins, in the olive flounder Paralichthys olivaceus. The coding region of PoEFEMP2 encodes a protein that contains six calcium-binding EGF-like (EGF-CA) domains and four complement Clr-like EGF-like (cEGF) domains. PoEFEMP2 shows 67.51-96.77 % similarities to orthologs in a variety of fish species. PoEFEMP2 mRNA was detected in all tissues examined; the highest levels of PoEFEMP2 mRNA expression were observed in the heart, testis, ovary and muscle. The PoEFEMP2 mRNA level increases during early development. In addition, the PoEFEMP2 mRNA level increased at 3 h post-infection (hpi) and decreased from 6 to 48 hpi in flounder Hirame natural embryo (HINAE) cells infected with viral hemorrhagic septicemia virus (VHSV). Disruption of PoEFEMP2 using the clustered regularly interspaced short palindromic repeats/CRISPR-associated-9 (CRISPR/Cas9) system resulted in a significant upregulation of VHSV G mRNA levels and immune-related genes expression in knockout cells. These findings implicate PoEFEMP2 in antiviral responses in P. olivaceus.

3.
Mycoses ; 67(5): e13748, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38783563

RESUMO

BACKGROUND: Dermatophytosis is a common and major public health concern worldwide. Despite the increasing availability of antifungal drugs, relapses and untreated cases of dermatophyte infections are reported. Therefore, novel antifungal agents are required. Aminopyrrolnitrin (APRN) shows promise for dermatophytosis treatment because of its antifungal activity. OBJECTIVES: This study aimed to assess the antifungal properties of APRN against Trichophyton verrucosum (T. verrucosum), in both laboratory settings and a guinea pig model. METHODS: The minimum inhibitory concentrations (MICs) of APRN and enilconazole against T. verrucosum were determined according to the CLSI M38 method. The skins of 16 male guinea pigs were infected with 1.0 × 108 conidia of T. verrucosum and the animals were grouped into sets of four: negative control group (NC) received normal saline; positive control group (PC) received 2 µg/mL of enilconazole; and APRN4 and APRN8 received 4 and 8 µg/mL of APRN, respectively. Clinical, mycological and histological efficacies were measured after 10 days. RESULTS: The MIC90 of APRN and enilconazole against T. verrucosum was 4 and 2 µg/mL, respectively. The clinical scores of PC, APRN4, and APRN8 were significantly lower than those of NC. Clinical and mycological efficacies were higher for APRN8, APRN4 and PC. No fungi were observed in the skin tissues of APRN4 and APRN8, while fungi were observed in 50% of the PC. CONCLUSION: APRN showed antifungal activity against T. verrucosum in vitro and in vivo and is a potential candidate for the treatment of dermatophytosis.


Assuntos
Antifúngicos , Modelos Animais de Doenças , Testes de Sensibilidade Microbiana , Tinha , Trichophyton , Animais , Cobaias , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Trichophyton/efeitos dos fármacos , Tinha/tratamento farmacológico , Tinha/microbiologia , Masculino , Pele/microbiologia
4.
Aging Male ; 26(1): 6-12, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36633207

RESUMO

INTRODUCTION: A positive association between benign prostate hyperplasia (BPH)/lower urinary tract symptoms (LUTS) and metabolic syndrome (MetS) was reported in several studies, but studies from Asia often showed conflicting results. MATERIALS AND METHODS: Medical records were obtained from a health promotion center database between 2021 and 2022. Men without a history of treatment for LUTS were evaluated using the International Prostate Symptom Score (IPSS), Overactive Bladder Symptom Score (OABSS), transrectal ultrasonography. RESULTS: Of 1345 individuals, 603 (44.8%) had MetS. Older age, higher IPSS values, higher prevalence rates of BPH and overactive bladder, higher triiodothyronine, and lower testosterone and sex-hormone binding globulin were observed in individuals with MetS than in individuals without MetS. The severity of LUTS significantly increased in the individuals with MetS (p = .002). In individuals with MetS, age, HbA1c, and cerebrovascular disease (CVD) were associated with IPSS. For OABSS, age, HbA1c, thyroid-stimulating hormone (TSH), coronary artery occlusive disease, and CVD were identified as predictors. CONCLUSIONS: We confirmed the positive correlation between MetS and BPH/LUTS in Korean. Factors including TSH and atherosclerosis affected LUTS in individuals with MetS. These findings suggested a potential role of thyroid hormones and atherosclerosis in the etiology and treatment of BPH/LUTS in patients with MetS.


Assuntos
Sintomas do Trato Urinário Inferior , Síndrome Metabólica , Hiperplasia Prostática , Humanos , Masculino , Doenças Cardiovasculares/epidemiologia , Hemoglobinas Glicadas/análise , Sintomas do Trato Urinário Inferior/epidemiologia , Síndrome Metabólica/epidemiologia , Hiperplasia Prostática/epidemiologia , República da Coreia/epidemiologia , Bexiga Urinária Hiperativa/epidemiologia
5.
Nano Lett ; 22(13): 5069-5076, 2022 07 13.
Artigo em Inglês | MEDLINE | ID: mdl-35648998

RESUMO

Structural colors have advantages compared with chemical pigments or dyes, such as iridescence, tunability, and unfading. Many studies have focused on developing the ability to switch ON/OFF the structural color; however, they often suffer from a simple and single stimulus, remaining structural colors, and target selectivity. Herein, we present regionally controlled multistimuli-responsive structural color switching surfaces. The key part is the utilization of a micropatterned DNA-hydrogel assembly on a single substrate. Each hydrogel network contains a unique type of stimuli-responsive DNA motifs as an additional cross-linker to exhibit swelling/deswelling via stimuli-responsive DNA interactions. The approach enables overcoming the existing limitations and selectively programming the DNA-hydrogel to a decrypted state (ON) and an encrypted state (OFF) in response to multiple stimuli. Furthermore, the transitions are reversible, providing cyclability. We envision the potential of our method for diverse applications, such as sensors or anticounterfeiting, requiring multistimuli-responsive structural color switching surfaces.


Assuntos
DNA , Hidrogéis , Corantes , DNA/química , Hidrogéis/química
6.
Int J Syst Evol Microbiol ; 72(12)2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36748413

RESUMO

A Gram-stain-negative, facultatively anaerobic, flagellated and coccoid, ovoid or rod-shaped bacterial strain, NIFS-20-8T, was isolated from the intestine of an olive flounder (Paralichthys olivaceus) from the East Sea, Republic of Korea. The neighbour-joining phylogenetic tree of 16S rRNA gene sequences showed that strain NIFS-20-8T fell within the clade comprising the type strains of Enterovibrio species. Strain NIFS-20-8T exhibited 16S rRNA gene sequence similarities of 97.2 and 97.1 % to the type strains of Enterovibrio nigricans and Enterovibrio norvegicus, respectively, and of 96.6-97.0 % to the type strains of the other Enterovibrio species. The average nucleotide identity and digital DNA-DNA hybridization values between the genomic sequence of strain NIFS-20-8T and those of the type strains of four Enterovibrio species were 73.8-75.0 and 19.8-21.1 %, respectively. The DNA G+C content of strain NIFS-20-8T from genomic sequence data was 50.55 mol%. Strain NIFS-20-8T contained Q-8 as the predominant ubiquinone and summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c), C16 : 0 and C18 : 1 ω7c as the major fatty acids. The major polar lipids detected in stain NIFS-20-8T were phosphatidylethanolamine and phosphatidylglycerol. Distinguishing phenotypic properties, together with phylogenetic and genetic distinctiveness, revealed that strain NIFS-20-8T is separated from recognized Enterovibrio species. On the basis of the data presented here, strain NIFS-20-8T is considered to represent a novel species of the genus Enterovibrio, for which the name Enterovibrio paralichthyis sp. nov. is proposed. The type strain is NIFS-20-8T (= KCTC 82873T=NBRC 115237T).


Assuntos
Ácidos Graxos , Linguado , Animais , Ácidos Graxos/química , Linguado/genética , Água do Mar/microbiologia , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , DNA Bacteriano/genética , Composição de Bases , Técnicas de Tipagem Bacteriana , República da Coreia
7.
Artigo em Inglês | MEDLINE | ID: mdl-35156920

RESUMO

A Gram-stain positive, aerobic, irregularly rod-shaped, non-spore-forming bacterium, designated as BN130099T, was isolated from farmland soil sampled in Goesan-gun, Chungbuk, Republic of Korea. Phylogenetic analysis of its 16S rRNA gene sequence showed that the strain is closely related to Nocardioides pelophilus KACC 19192T with 98.11 % similarity. The DNA G+C content of strain BN130099T was 68.84 mol% (draft genome sequence). The genome sequence of BN130099T displayed key enzymes involved in bioremediation of organic pollutants and biosynthetic clusters of saquayamycin. The strain contained ll-2,6-diaminopimelic acid in the cell-wall peptidoglycan and MK-8(H4) as the major respiratory quinone. The predominant fatty acid was iso-C16 : 0. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine and phosphatidylinositol. The results of physiological and biochemical characterization allowed the phenotypic differentiation of strain BN130099T from N. pelophilus KACC 19192T. The strain represents a novel species of the genus Nocardioides, for which we propose the name Nocardioides humilatus sp. nov. The type strain is BN130099T (=KCTC 49079T=CCTCC AB 2018135T).


Assuntos
Nocardioides/classificação , Filogenia , Microbiologia do Solo , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Fazendas , Ácidos Graxos/química , Nocardioides/isolamento & purificação , Fosfolipídeos/química , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Vitamina K 2/análogos & derivados , Vitamina K 2/química
8.
Fish Shellfish Immunol ; 127: 666-671, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35803510

RESUMO

Tripartite motif-containing (TRIM) proteins are conserved throughout the metazoan kingdom, and the TRIM subset finTRIM is highly diversified in fish. We isolated TRIM16 cDNA, a member of the finTRIM family, from the olive flounder Paralichthys olivaceus (PoTRIM16). PoTRIM16 contained a 1,725-bp coding sequence encoding a 574-amino acid polypeptide, which in turn contained a really interesting new gene (RING) finger domain, B-box-type zinc finger (B-BOX), nuclease SbcCD subunit C (SbcC), structural maintenance of chromosome (SMC prok B), and stonustoxin (SNTX) subunit alpha (SPRY-PRY-SNTX). Multiple alignment of related sequences revealed that PoTRIM16 showed 86.63-97.40% identity with fish orthologues, and a phylogenetic tree was constructed of vertebrates. PoTRIM16 mRNA was detected in all tissues examined; levels were highest in the eye and ovary. PoTRIM16 mRNA expression was investigated during early development. Under VHSV infection, PoTRIM16 mRNA was downregulated in the liver of P. olivaceus. This is the first study to characterize fish-specific finTRIM in P. olivaceus, which may play a role in the immune response against virus infection.


Assuntos
Doenças dos Peixes , Linguado , Novirhabdovirus , Animais , Feminino , Novirhabdovirus/fisiologia , Filogenia , RNA Mensageiro/metabolismo
9.
Cell Mol Life Sci ; 78(1): 207-225, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32140747

RESUMO

NAD(P)-dependent steroid dehydrogenase-like (NSDHL), an essential enzyme in human cholesterol synthesis and a regulator of epidermal growth factor receptor (EGFR) trafficking pathways, has attracted interest as a therapeutic target due to its crucial relevance to cholesterol-related diseases and carcinomas. However, the development of pharmacological agents for targeting NSDHL has been hindered by the absence of the atomic details of NSDHL. In this study, we reported two X-ray crystal structures of human NSDHL, which revealed a detailed description of the coenzyme-binding site and the unique conformational change upon the binding of a coenzyme. A structure-based virtual screening and biochemical evaluation were performed and identified a novel inhibitor for NSDHL harboring suppressive activity towards EGFR. In EGFR-driven human cancer cells, treatment with the potent NSDHL inhibitor enhanced the antitumor effect of an EGFR kinase inhibitor. Overall, these findings could serve as good platforms for the development of therapeutic agents against NSDHL-related diseases.


Assuntos
3-Hidroxiesteroide Desidrogenases/metabolismo , Inibidores Enzimáticos/metabolismo , 3-Hidroxiesteroide Desidrogenases/antagonistas & inibidores , 3-Hidroxiesteroide Desidrogenases/química , 3-Hidroxiesteroide Desidrogenases/genética , Sítios de Ligação , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Colesterol/química , Cristalografia por Raios X , Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , Receptores ErbB/antagonistas & inibidores , Receptores ErbB/metabolismo , Cloridrato de Erlotinib/química , Cloridrato de Erlotinib/metabolismo , Cloridrato de Erlotinib/farmacologia , Humanos , Cinética , Simulação de Acoplamento Molecular , Mutagênese Sítio-Dirigida , NAD/química , NAD/metabolismo , Estrutura Terciária de Proteína , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/isolamento & purificação , Transdução de Sinais
10.
BMC Urol ; 22(1): 164, 2022 Oct 29.
Artigo em Inglês | MEDLINE | ID: mdl-36309672

RESUMO

BACKGROUND: To analyze grayscale values for hypoechoic lesions matched with target lesions evaluated using prebiopsy magnetic resonance imaging (MRI) according to the Prostate Imaging-Reporting and Data System (PI-RADS). METHODS: We collected data on 420 target lesions in patients who underwent MRI/transrectal ultrasound fusion-targeted biopsies between January 2017 and September 2020. Images of hypoechoic lesions that matched the target lesions on MRI were stored in a picture archiving and communication system, and their grayscale values were estimated using the red/green/blue scoring method through an embedded function. We analyzed imaging data using grayscale values. RESULTS: Of the 420 lesions, 261 (62.1%) were prostate cancer lesions. There was no difference in the median grayscale values between benign and prostate cancer lesions. However, grayscale ranges (41.8-98.5 and 42.6-91.8) were significant predictors of prostate cancer and clinically significant prostate cancer (csPC) in multivariable logistic regression analyses. Area under the curve for detecting csPC using grayscale values along with conventional variables (age, prostate-specific antigen levels, prostate volume, previous prostate biopsy results, and PI-RADS scores) was 0.839, which was significantly higher than that for detecting csPC using only conventional variables (0.828; P = 0.036). Subgroup analysis revealed a significant difference for PI-RADS 3 lesions between grayscale values for benign and cancerous lesions (74.5 vs. 58.8, P = 0.008). Grayscale values were the only significant predictive factor (odds ratio = 4.46, P = 0.005) for csPC. CONCLUSIONS: Distribution of grayscale values according to PI-RAD 3 scores was potentially useful, and the grayscale range (42.6-91.8) was a potential predictor for csPC diagnosis.


Assuntos
Neoplasias da Próstata , Masculino , Humanos , Neoplasias da Próstata/patologia , Imageamento por Ressonância Magnética/métodos , Gradação de Tumores , Estudos Retrospectivos , Biópsia Guiada por Imagem/métodos
11.
Sensors (Basel) ; 22(12)2022 Jun 16.
Artigo em Inglês | MEDLINE | ID: mdl-35746337

RESUMO

This paper presents an on-chip implementation of an analog processor-in-memory (PIM)-based convolutional neural network (CNN) in a biosensor. The operator was designed with low power to implement CNN as an on-chip device on the biosensor, which consists of plates of 32 × 32 material. In this paper, 10T SRAM-based analog PIM, which performs multiple and average (MAV) operations with multiplication and accumulation (MAC), is used as a filter to implement CNN at low power. PIM proceeds with MAV operations, with feature extraction as a filter, using an analog method. To prepare the input feature, an input matrix is formed by scanning a 32 × 32 biosensor based on a digital controller operating at 32 MHz frequency. Memory reuse techniques were applied to the analog SRAM filter, which is the core of low power implementation, and in order to accurately grasp the MAC operational efficiency and classification, we modeled and trained numerous input features based on biosignal data, confirming the classification. When the learned weight data was input, 19 mW of power was consumed during analog-based MAC operation. The implementation showed an energy efficiency of 5.38 TOPS/W and was differentiated through the implementation of 8 bits of high resolution in the 180 nm CMOS process.


Assuntos
Técnicas Biossensoriais , Redes Neurais de Computação , Aprendizagem
12.
Int J Syst Evol Microbiol ; 71(11)2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34752209

RESUMO

A Gram-stain-negative, strictly aerobic, non-motile and rod-shaped bacterial strain, MYP1-1T, was isolated from the intestine of a stalked sea squirt (Styela clava) of the South Sea in the Republic of Korea. The neighbour-joining phylogenetic tree based on 16S rRNA gene sequences revealed that strain MYP1-1T clustered with the type strains of Halocynthiibacter species and Pseudohalocynthiibacter aestuariivivens. Strain MYP1-1T exhibited 16S rRNA gene sequence similarity values of 97.0-97.6 % to the type strains of Halocynthiibacter namhaensis, Halocynthiibacter arcticus and P. aestuariivivens. The phylogenetic tree based on genomic sequences showed that strain MYP1-1T formed a distinct branch separating it from the type strains of two Halocynthiibacter species and P. aestuariivivens and other taxa. The DNA G+C content of strain MYP1-1T from its genomic sequence was 55.0 mol%. Strain MYP1-1T contained Q-10 as the predominant ubiquinone and C18 : 1 ω7c as the major fatty acid. The major polar lipids of strain MYP1-1T were phosphatidylcholine, phosphatidylglycerol, one unidentified lipid and one unidentified aminolipid. The differences in fatty acid and polar lipid profiles and other differential phenotypic properties made it reasonable to distinguish strain MYP1-1T from the genera Halocynthiibacter and Pseudohalocynthiibacter. On the basis of the polyphasic taxonomic investigations, we conclude that strain MYP1-1T constitutes a new genus and species within the class Alphaproteobacteria, for which the name Paenihalocynthiibacter styelae gen. nov., sp. nov. is proposed. The type strain is MYP1-1T (=KCTC 82143T=NBRC 114355T).


Assuntos
Filogenia , Rhodobacteraceae , Urocordados , Animais , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Fosfolipídeos/química , RNA Ribossômico 16S/genética , República da Coreia , Rhodobacteraceae/classificação , Rhodobacteraceae/isolamento & purificação , Água do Mar , Análise de Sequência de DNA , Ubiquinona/análogos & derivados , Ubiquinona/química , Urocordados/microbiologia
13.
Mar Drugs ; 19(8)2021 Aug 06.
Artigo em Inglês | MEDLINE | ID: mdl-34436290

RESUMO

American oyster defensin (AOD) was previously purified from acidified gill extract of the American oyster, Crassostrea virginica. AOD is composed of 38 amino acids with three disulfide bonds and exhibits strong antimicrobial activity against Gram-positive bacteria as well as significant activity against Gram-negative bacteria. Here, to develop promising peptides into antibiotic candidates, we designed five arginine-rich analogs (A0, A1, A2, A3, and A4), predicted their loop and extended strand/random structures-including nine amino acids and a disulfide bond derived from the C-terminus of AOD-and described their antimicrobial and cytotoxic effects, as well as their modes of action. In our experimental results, the A3 and A4 analogs exhibited potent antimicrobial activity against all test organisms-including four Gram-positive bacteria, six Gram-negative bacteria, and Candida albicans-without cell toxicity. A sequence of experiments, including a membrane permeabilization assay, DNA binding study, and DNA polymerization inhibition test, indicated that the two analogs (A3 and A4) possibly did not act directly on the bacterial membrane but instead interacted with intracellular components such as DNA or DNA amplification reactions. AOD analogs also showed strong bacterial inhibition activity in the plasma environment. In addition, analog-treated microbial cells clearly exhibited membrane disruption, damage, and leakage of cytoplasmic contents. Collectively, our results suggest that two analogs, A3 and A4, have potent antimicrobial activity via DNA interaction and have the potential for development into novel antimicrobial agents.


Assuntos
Antibacterianos/farmacologia , Defensinas/farmacologia , Ostreidae , Animais , Organismos Aquáticos , Eritrócitos/efeitos dos fármacos , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Hemólise/efeitos dos fármacos , Humanos , Testes de Sensibilidade Microbiana , Fitoterapia
14.
Int J Mol Sci ; 22(3)2021 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-33572970

RESUMO

MAF1 is a global suppressor of RNA polymerase III-dependent transcription, and is conserved from yeast to human. Growing evidence supports the involvement of MAF1 in the immune response of mammals, but its biological functions in fish are unknown. We isolated and characterized Maf1 from the olive flounder Paralichthys olivaceus (PoMaf1). The coding region of PoMaf1 comprised 738 bp encoding a 245-amino-acid protein. The deduced PoMAF1 amino acid sequence shared features with those of MAF1 orthologues from vertebrates. PoMaf1 mRNA was detected in all tissues examined, and the levels were highest in eye and muscle tissue. The PoMaf1 mRNA level increased during early development. In addition, the PoMaf1 transcript level decreased during viral hemorrhagic septicemia virus (VHSV) infection of flounder hirame natural embryo (HINAE) cells. To investigate the role of PoMaf1 in VHSV infection, single-cell-derived PoMaf1 knockout HINAE cells were generated using the clustered regularly interspaced short palindromic repeats/CRISPR-associated-9 (CRISPR/Cas9) system, and cell clones with complete disruption of PoMaf1 were selected. PoMaf1 disruption increased the VHSV glycoprotein (G) mRNA levels during VHSV infection of HINAE cells, implicating PoMAF1 in the immune response to VSHV infection. To our knowledge, this is the first study to characterize fish Maf1, which may play a role in the response to viral infection.


Assuntos
Doenças dos Peixes/genética , Proteínas de Peixes/genética , Linguado/genética , Septicemia Hemorrágica/veterinária , Novirhabdovirus/fisiologia , Proteínas Repressoras/genética , Animais , Sistemas CRISPR-Cas , Linhagem Celular , Doenças dos Peixes/imunologia , Proteínas de Peixes/imunologia , Linguado/imunologia , Linguado/fisiologia , Septicemia Hemorrágica/genética , Septicemia Hemorrágica/imunologia , Interações Hospedeiro-Patógeno , Novirhabdovirus/imunologia , Filogenia , Proteínas Repressoras/imunologia , Transcrição Gênica
15.
Biochem Biophys Res Commun ; 527(1): 264-269, 2020 06 18.
Artigo em Inglês | MEDLINE | ID: mdl-32446378

RESUMO

Toxin-antitoxin (TA) systems are ubiquitously found in bacteria and are related to cell maintenance and survival under environmental stresses such as heat shock, nutrient starvation, and antibiotic treatment. Here, we report for the first time the crystal structure of the Staphylococcus aureus TA complex YoeBSa1-YefMSa1 at a resolution of 1.7 Å. This structure reveals a heterotetramer with a 2:2 stoichiometry between YoeBSa1 and YefMSa1. The N-terminal regions of the YefMSa1 antitoxin form a homodimer characteristic of a hydrophobic core, and the C-terminal extended region of each YefMSa1 protomer makes contact with each YoeBSa1 monomer. The binding stoichiometry of YoeBSa1 and YefMSa1 is different from that of YoeB and YefM of E. coli (YoeBEc and YefMEc), which is the only structural homologue among YoeB-YefM families; however, the structures of individual YoeBSa1 and YefMSa1 subunits in the complex are highly similar to the corresponding structures in E. coli. In addition, docking simulation with a minimal RNA substrate provides structural insight into the guanosine specificity of YoeBSa1 for cleavage in the active site, which is distinct from the specificity of YoeBEc for adenosine rather than guanosine. Given the previous finding that YoeBSa1 exhibits fatal toxicity without inducing persister cells, the structure of the YoeBSa1-YefMSa1 complex will contribute to the design of a new category of anti-staphylococcal agents that disrupt the YoeBSa1-YefMSa1 complex and increase YoeBSa1 toxicity.


Assuntos
Proteínas de Bactérias/química , Toxinas Bacterianas/química , Endorribonucleases/química , Staphylococcus aureus/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/isolamento & purificação , Toxinas Bacterianas/genética , Toxinas Bacterianas/isolamento & purificação , Endorribonucleases/genética , Endorribonucleases/isolamento & purificação , Simulação de Acoplamento Molecular , Conformação Proteica
16.
Biochem Biophys Res Commun ; 532(2): 173-178, 2020 11 05.
Artigo em Inglês | MEDLINE | ID: mdl-32838967

RESUMO

Acylphosphatase is the smallest enzyme that is widely distributed in many diverse organisms ranging from archaebacteria to higher-eukaryotes including the humans. The enzyme hydrolyzes the carboxyl-phosphate bonds of the acyl phosphates which are important intermediates in glycolysis, membrane pumps, tricarboxylic acid cycle, and urea biosynthesis. Despite its biological importance in critical cellular functions, very limited structural investigations have been conducted on bacterial acylphosphatases. Here, we first unveiled the crystal structure of SaAcP, an acylphosphatase from gram-positive S. aureus at the atomic level. Structural insights on the active site together with mutation study provided greater understanding of the catalytic mechanism of SaAcP as a bacterial acylphosphatase and as a putative apyrase. Furthermore, through NMR titration experiment of SaAcP in its solution state, the dynamics and the alterations of residues affected by the phosphate ion were validated. Our findings elucidate the structure-function relationship of acylphosphatases in gram-positive bacteria and will provide a valuable basis for researchers in the field related to bacterial acylphosphatases.


Assuntos
Hidrolases Anidrido Ácido/química , Hidrolases Anidrido Ácido/metabolismo , Staphylococcus aureus/enzimologia , Hidrolases Anidrido Ácido/genética , Trifosfato de Adenosina/metabolismo , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Benzoatos/metabolismo , Domínio Catalítico , Cristalografia por Raios X , Espectroscopia de Ressonância Magnética , Modelos Moleculares , Conformação Proteica , Acilfosfatase
17.
Fish Shellfish Immunol ; 99: 342-352, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32061872

RESUMO

We isolated and purified an antimicrobial peptide (AMP) from the mantle of the hard-shelled mussel, Mytilus coruscus. The peptide was purified through C18 reversed-phase high-performance liquid chromatography, and displayed antibacterial activity. Total molecular mass of 11,182 Da was determined using matrix-assisted laser desorption ionization time-of-flight mass spectrophotometry. The N-terminal 23-amino acid sequence of its purified peak was obtained through Edman degradation, revealing 82% identity with myticusin-1 of M. coruscus. Complete sequence of the target peptide was determined through cDNA cloning and rapid amplification of cDNA ends. The complete sequence comprised 574 bp with a 387-bp open reading frame (ORF) encoding 24 amino acids of a signal peptide and 104 amino acids of a mature peptide, which was named myticusin-beta. Furthermore, we discovered two novel isoforms of myticusin-beta. We constructed and expressed recombinant myticusin-beta, which displayed antimicrobial activity against gram-positive (Bacillus cereus, Bacillus subtilis, Clostridium perfringens, Staphylococcus aureus, Streptococcus iniae, Streptococcus mutans) and gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa, Vibrio alginolyticus, Klebsiella pneumoniae). Purified recombinant myticusin-beta also showed anti-parasitic activity at various concentrations. A short AMP analog was designed and synthesized based on the sequence of myticusin-beta, with markedly improved antimicrobial activity. Expression of myticusin-beta was detected in the mantle at the highest level, followed by hemocytes. The results obtained in this work suggest that myticusin-beta is an immune-related AMP of M. coruscus and an effective alternative to antibiotics.


Assuntos
Peptídeos Catiônicos Antimicrobianos/metabolismo , Peptídeos Catiônicos Antimicrobianos/farmacologia , Mytilus/metabolismo , Animais , Peptídeos Catiônicos Antimicrobianos/química , Bactérias/efeitos dos fármacos , Candida albicans/efeitos dos fármacos , Humanos
18.
Macromol Rapid Commun ; 41(7): e2000013, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32068937

RESUMO

Thermadapt shape memory polymers (SMPs), utilizing a variety of dynamic covalent bond exchange mechanisms, have been extensively studied in recent years but it is still challenging to address several constraints in terms of limited accuracy and complexity for constructing 3D shape memory structures. Here, an effective and facile preparation of thermadapt SMPs based on elemental sulfur-derived poly(phenylene polysulfide) networks (PSNs) is presented. These SMPs possess intrinsic near-infrared (NIR)-induced photothermal conversion properties for spatiotemporal control of their plasticity and elasticity. The NIR-controllable plasticity and elasticity of the PSNs enable versatile shape manipulation of 3D multi-shape memory structures, including building block assembly, reconfiguration, shape fixing/recovery, and repair.


Assuntos
Materiais Biocompatíveis/química , Sulfetos/química , Enxofre/química , Elasticidade , Raios Infravermelhos , Teste de Materiais , Estrutura Molecular , Tamanho da Partícula
19.
Int J Syst Evol Microbiol ; 69(9): 2781-2786, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31246165

RESUMO

A Gram-stain-negative, aerobic, non-motile and ovoid or rod-shaped bacterial strain, MRS2T, was isolated from an intestine of Nile tilapia (Oreochromisniloticus) collected from the Republic of Korea. Strain MRS2T grew optimally at 30 °C and in the presence of 0-2.0 % (w/v) NaCl. A neighbour-joining phylogenetic tree of 16S rRNA gene sequences showed that strain MRS2T clustered with the type strains of Empedobacter species. It exhibited the highest 16S rRNA gene sequence similarity (98.5 %) to the type strain of Empedobacter falsenii and sequence similarities of 97.4-97.6 % to the type strains of two other Empedobacter species. Strain MRS2T contained MK-6 as the predominant ubiquinone and summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c), iso-C17 : 0 3-OH and iso-C15 : 0 as the major fatty acids. The major polar lipids of strain MRS2T were phosphatidylethanolamine, one unidentified lipid and one unidentified aminolipid. The DNA G+C contents of strain MRS2T were 32.2 mol% or 30.65 mol%. Strain MRS2T exhibited DNA-DNA relatedness values of 12-20 % to the type strains of Empedobacter falsenii, Empedobacter brevis and Empedobacter stercoris. The average nucleotide identity values between strain MRS2T and five strains of E. falsenii and E. brevis were 84.8-91.0 %. The phylogenetic, genetic and differential phenotypic properties indicated that strain MRS2T is separated from Empedobacter species. On the basis of the data presented here, strain MRS2T is considered to represent a novel species of the genus Empedobacter, for which the name Empedobactertilapiae sp. nov. is proposed. The type strain is MRS2T (=KCTC 62904T=NBRC 113550T).


Assuntos
Ciclídeos/microbiologia , Flavobacteriaceae/classificação , Intestinos/microbiologia , Filogenia , Animais , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Flavobacteriaceae/isolamento & purificação , Flavobacterium/genética , Fosfatidiletanolaminas/química , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Vitamina K 2/análogos & derivados , Vitamina K 2/química
20.
Int J Syst Evol Microbiol ; 69(10): 3109-3115, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31334695

RESUMO

A Gram-stain-negative, aerobic, non-motile and coccoid, ovoid or rod-shaped bacterial strain, designated MYP11T, was isolated from seawater around Jeju island, Republic of Korea and identified by polyphasic taxonomic study. A neighbour-joining phylogenetic tree of 16S rRNA gene sequences showed that strain MYP11T joined the cluster comprising the type strains of Shimiaabyssi, Shimiaaestuarii and Shimiaaquaeponti, showing 16S rRNA gene sequence similarities of 96.3-96.8 %. Strain MYP11T exhibited 16S rRNA gene sequence similarity values of 94.2-94.9 % to the type strains of other Shimia species. In the upgma dendrogram based on the average nucleotide identity values of genomic sequences, strain MYP11T formed an evolutionary lineage independent of those of Shimia species and other taxa. Strain MYP11T contained Q-10 as the predominant ubiquinone and C18 : 1 ω7c and cyclo C19 : 0 ω8c as the major fatty acids. The major polar lipids of strain MYP11T were phosphatidylcholine, phosphatidylglycerol, two unidentified lipids and one unidentified aminolipid. The DNA G+C content of strain MYP11T was 63.1 or 61.5 mol%. The differences in the fatty acid and polar lipid profiles and DNA G+C contents made it reasonable to distinguish strain MYP11T from the type strains of S. abyssi, S. aestuarii and S. aquaeponti and those of other Shimia species. On the basis of the polyphasic data presented here, strain MYP11T is considered to constitute a new genus and species within the class Alphaproteobacteria, for which the name Aliishimia ponticola gen. nov., sp. nov. is proposed. The type strain is MYP11T (=KCTC 62899T=NBRC 113544T).


Assuntos
Filogenia , Rhodobacteraceae/classificação , Água do Mar/microbiologia , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Fosfolipídeos/química , RNA Ribossômico 16S/genética , República da Coreia , Rhodobacteraceae/isolamento & purificação , Análise de Sequência de DNA , Ubiquinona/análogos & derivados , Ubiquinona/química
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