RESUMO
Elevated intraocular pressure (IOP) in glaucoma causes retinal ganglion cell (RGC) loss and damage to the optic nerve. Although IOP is controlled pharmacologically, no treatment is available to restore retinal and optic nerve function. In this paper, we aimed to develop a novel gene therapy for glaucoma using an AAV2-based thioredoxin 2 (Trx2)-exoenzyme C3 transferase (C3) fusion protein expression vector (scAAV2-Trx2-C3). We evaluated the therapeutic effects of this vector in vitro and in vivo using dexamethasone (DEX)-induced glaucoma models. We found that scAAV2-Trx2-C3-treated HeLa cells had significantly reduced GTP-bound active RhoA and increased phosphor-cofilin Ser3 protein expression levels. scAAV2-Trx2-C3 was also shown to inhibit oxidative stress, fibronectin expression, and alpha-SMA expression in DEX-treated HeLa cells. NeuN immunostaining and TUNEL assay in mouse retinal tissues was performed to evaluate its neuroprotective effect upon RGCs, whereas changes in mouse IOP were monitored via rebound tonometer. The present study showed that scAAV2-Trx2-C3 can protect RGCs from degeneration and reduce IOP in a DEX-induced mouse model of glaucoma, while immunohistochemistry revealed that the expression of fibronectin and alpha-SMA was decreased after the transduction of scAAV2-Trx2-C3 in murine eye tissues. Our results suggest that AAV2-Trx2-C3 modulates the outflow resistance of the trabecular meshwork, protects retinal and other ocular tissues from oxidative damage, and may lead to the development of a gene therapeutic for glaucoma.
Assuntos
Glaucoma , Pressão Intraocular , Humanos , Camundongos , Animais , Células Ganglionares da Retina/metabolismo , Fibronectinas/metabolismo , Tiorredoxinas/metabolismo , Células HeLa , Transferases/metabolismo , Glaucoma/genética , Glaucoma/terapia , Glaucoma/metabolismo , Modelos Animais de DoençasRESUMO
In addition to laser photocoagulation, therapeutic interventions for diabetic retinopathy (DR) have heretofore consisted of anti-VEGF drugs, which, besides drawbacks inherent to the treatments themselves, are limited in scope and may not fully address the condition's complex pathophysiology. This is because DR is a multifactorial condition, meaning a gene therapy focused on a target with broader effects, such as the mechanistic target of rapamycin (mTOR), may prove to be the solution in overcoming these concerns. Having previously demonstrated the potential of a mTOR-inhibiting shRNA packaged in a recombinant adeno-associated virus to address a variety of angiogenic retinal diseases, here we explore the effects of rAAV2-shmTOR-SD in a streptozotocin-induced diabetic mouse model. Delivered via intravitreal injection, the therapeutic efficacy of the virus vector upon early DR processes was examined. rAAV2-shmTOR-SD effectively transduced mouse retinas and therein downregulated mTOR expression, which was elevated in sham-treated and control shRNA-injected (rAAV2-shCon-SD) control groups. mTOR inhibition additionally led to marked reductions in pericyte loss, acellular capillary formation, vascular permeability, and retinal cell layer thinning, processes that contribute to DR progression. Immunohistochemistry showed that rAAV2-shmTOR-SD decreased ganglion cell loss and pathogenic Müller cell activation and proliferation, while also having anti-apoptotic activity, with these effects suggesting the therapeutic virus vector may be neuroprotective. Taken together, these results build upon our previous work to demonstrate the broad ability of rAAV2-shmTOR-SD to address aspects of DR pathophysiology further evidencing its potential as a human gene therapeutic strategy for DR.
Assuntos
Diabetes Mellitus , Retinopatia Diabética , Animais , Dependovirus/genética , Diabetes Mellitus/metabolismo , Retinopatia Diabética/genética , Retinopatia Diabética/metabolismo , Retinopatia Diabética/terapia , Vetores Genéticos/genética , Camundongos , RNA Interferente Pequeno/metabolismo , Retina/patologia , Serina-Treonina Quinases TOR/genética , Serina-Treonina Quinases TOR/metabolismoRESUMO
Expanding on previous demonstrations of the therapeutic effects of adeno-associated virus (AAV) carrying small-hairpin RNA (shRNA) in downregulating the mechanistic target of rapamycin (mTOR) in in vivo retinal vascular disorders, vascular endothelial growth factor (VEGF)-stimulated endothelial cells were treated with AAV2-shmTOR to examine the role of mTOR inhibition in retinal angiogenesis. AAV2-shmTOR exposure significantly reduced mTOR expression in human umbilical vein endothelial cells (HUVECs) and decreased downstream signaling cascades of mTOR complex 1 (mTORC1) and mTORC2 under VEGF treatment. Moreover, the angiogenic potential of VEGF was significantly inhibited by AAV2-shmTOR, which preserved endothelial integrity by maintaining tight junctions between HUVECs. These data thus support previous in vivo studies and provide evidence that AAV2-shmTOR induces therapeutic effects by inhibiting the neovascularization of endothelial cells.
Assuntos
Dependovirus , Fator A de Crescimento do Endotélio Vascular , Dependovirus/genética , Dependovirus/metabolismo , Células Endoteliais da Veia Umbilical Humana , Humanos , RNA Interferente Pequeno/farmacologia , Serina-Treonina Quinases TOR/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
Androgen exerts its functions by binding with an androgen receptor (AR). It can activate many signaling pathways that are important to the progression of castration-resistant prostate cancer (CRPC). Here, we characterized the rapid proteomic changes seen at 5, 15, 30, and 60 min after the androgen treatment of VCaP cells via the tandem mass tag (TMT) labeling strategy. A total of 5529 proteins were successfully identified and quantified. Dynamic time profiling of protein expression patterns allowed us to identify five protein clusters involved in various stages of androgen-initiated signal transmission and processing. More details of protein functions and localization patterns, and our elucidation of an AR-interacting protein network, were obtained. Finally, we validated the expression level of AR-regulated proteins known to be significantly regulated in CRPC patients using the mouse xenograft model and patient samples. Our work offers a systematic analysis of the rapid proteomic changes induced by androgen and provides a global view of the molecular mechanisms underlying CRPC progression.
RESUMO
Strain U15T, rod-shaped, catalase and oxidase positive, non-motile, hot pink pigmented, Gram-negative bacterium, was isolated from soil of Udo port, Udo Island, South Korea. Growth was observed at 10-48 °C, pH 6-11, and 0% (w/v) NaCl. Optimum growth conditions are 30-40 °C, pH 7-10, and 0% (w/v) NaCl. Phylogenetic analysis based on 16S rRNA sequences showed that strain U15T forms a distinct clade with type strains of the family Cytophagaceae, with similarities below 89%. The major polar lipids are phosphatidylethanolamine and phosphatidylserine. Strain U15T was found to contain MK-7 as the only menaquinone, and iso-C15:0, C16:1ω5c, iso-C17:0 3-OH and summed feature 3 (comprising C16:1ω7c and/or C16:1ω6c) as the major fatty acids (> 10%). The DNA G + C content of strain U15T was determined to be 54.3 mol%. Based on the phylogenetic, phenotypic characteristics and chemotaxonomic analysis data, strain U15T (= KCTC 62116T = JCM 32361T) should be classified as representing a novel species of a novel genus within the family Cytophagaceae for which the name Tellurirhabdus rosea gen. nov., sp. nov., is proposed.
Assuntos
Cytophagaceae/isolamento & purificação , Microbiologia do Solo , Técnicas de Tipagem Bacteriana , Composição de Bases , Cytophagaceae/classificação , Cytophagaceae/genética , Cytophagaceae/metabolismo , DNA Bacteriano/genética , Ácidos Graxos/química , Ácidos Graxos/metabolismo , Filogenia , RNA Ribossômico 16S/genética , República da CoreiaRESUMO
A short rod-shaped, yellow-orange pigmented, strictly aerobic bacterium, designated as strain H2T, was isolated from the wetland soil of Halla Mountain, Jeju-island, South Korea. Growth was observed at temperatures of 10-30 °C (optimum at 25-30 °C), pH of 6-8 (optimum at pH 7), and salt concentrations of 0-1% (w/v) NaCl (optimum at 0%). The strain H2T was found to be a catalase and oxidase-positive, non-motile, Gram-negative bacterium. On the basis of 16S rRNA gene sequence similarity and phylogenetic analysis, strain H2T was found to be related to the members of the Chitinophagaceae family, being closely related to Taibaiella chishuiensis AY17T (94.3% sequence similarity). The major polar lipids are phosphatidylethanolamine and glycolipid. Strain H2T contained MK-7 as the only menaquinone as well as iso-C15:0, iso-C15:1 G and iso-C17:0 3-OH as the major fatty acids (> 15%). The DNA G+C content of strain H2T was determined to be 48.3 mol%. Based on the phylogenetic, phenotypic characteristics and chemotaxonomic analysis data, strain H2T (= KCTC 62115T = JCM 32353T) should be classified as representative of a novel species of a novel genus within the family Chitinophagaceae, for which the name Edaphocola aurantiacus gen. nov., sp. nov., is proposed.
Assuntos
Bacteroidetes/isolamento & purificação , Microbiologia do Solo , Técnicas de Tipagem Bacteriana , Bacteroidetes/classificação , Bacteroidetes/genética , Bacteroidetes/metabolismo , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Ácidos Graxos/metabolismo , Filogenia , RNA Ribossômico 16S/genética , República da Coreia , Solo/química , Áreas AlagadasRESUMO
A Gram-negative, non-motile, aerobic bacterium, designated strain IP7T, was isolated from seawater at the shore of the Incheon Eulwang-ri beach, South Korea. Cells of strain IP7T are straight or slightly rod-shaped and colonies are round, convex and orange-yellow. Strain IP7T is flexirubin-negative, mild halophile, catalase-and oxidase-positive, and produces a yellow-orange carotenoid pigment. Growth is optimal at 30°C, pH 7-9, and 2.0-4.0% NaCl (w/v). On the basis of 16S rRNA gene sequence similarity, strain IP7T is affiliated with genus Aestuariibaculum in the family Flavobacteriaceae, the closest relative being Aestuariibaculum suncheonense SC17T (98.3% sequence similarity). The DNA G + C content of the novel strain is 37.4 mol%. The only quinone is MK-6 menaquinone. Iso-branched C15:0, iso-branched C15:1 G, and iso-branched C17:0 3-OH are major fatty acids. The major polar lipids are phosphatidylethanolamine, an unidentified aminoglycolipid and two unidentified glycolipids. The DNA-DNA hybridization value of strain IP7T with Aestuariibaculum suncheonense SC17T is 28.87%. Based on the collective DNA-DNA hybridization, biochemical, phylogenetic and physiological data, we report a novel species of the genus Aestuariibaculum for which the name Aestuariibaculum marinum sp. nov. is proposed. The type strain is IP7T (= KCTC 52521T = JCM 31725T).
Assuntos
Flavobacteriaceae/classificação , Flavobacteriaceae/isolamento & purificação , Filogenia , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano , Ácidos Graxos/análise , Flavobacteriaceae/genética , Flavobacteriaceae/fisiologia , Hibridização de Ácido Nucleico , Fosfatidiletanolaminas/análise , Pigmentação , RNA Ribossômico 16S/genética , República da Coreia , Água do Mar/microbiologia , Especificidade da Espécie , Vitamina K 2/análogos & derivadosRESUMO
A novel Gram-negative, orange pigmented, strictly aerobic bacterium, designated strain IP9T, was isolated from seawater at the sea shore of Incheon Eulwang-ri beach, South Korea. Cells of strain IP9T were observed to be straight or slightly curved rods and colonies to be round and convex. Strain IP9T was found to be catalase and oxidase positive, and non-motile. Growth was observed in the temperature range of 10-37 °C (optimum at 30 °C), pH range of 6-10 (optimum at pH 7-8) and salt concentration range of 0-7% (w/v) NaCl (optimum at 0-1%). On the basis of 16S rRNA gene sequence similarity and phylogenetic analysis, strain IP9T was found to be related to the members of the family Flavobacteriaceae, being closely related to Hwangdonia seohaensis KCTC 32177T (95.3% sequence similarity). The DNA G + C content of the novel strain was determined to be 39.1 mol%. The major polar lipids were found to be phosphatidylethanolamine, three unidentified aminoglycolipids and two unidentified glycolipids. The major fatty acids (> 10%) were identified as iso-C15:0 and iso-C17:0 3-OH. The predominant quinone was found to be menaquinone 6 (MK-6). Based on the biochemical, phylogenetic and physiological data, we conclude that strain IP9T (= KCTC 52523T = JCM 31732T) represents the type species of a novel genus of the family Flavobacteriaceae for which the name Thalassorhabdus aurantiaca gen. nov., sp. nov. is proposed.
Assuntos
Flavobacteriaceae/metabolismo , Água do Mar/microbiologia , Composição de Bases/genética , Flavobacteriaceae/genética , Fosfatidiletanolaminas/metabolismo , Filogenia , RNA Ribossômico 16S/genética , República da Coreia , TemperaturaRESUMO
A novel Gram-stain-negative bacterium, designated strain S8T, was isolated from a soil sample obtained in Gyeonggi Province, Republic of Korea. Cells of strain S8T were endospore-forming, motile by means of peritrichous flagella, and rod-shaped. S8T colonies were round, convex, wavy and white. Strain S8T grew optimally at 37 °C, pH 6-8, and up to 2.0â% (w/v) NaCl. Based on 16S rRNA gene sequence similarity, strain S8T was affiliated with the genus Paenibacillus in the family Paenibacillaceae and was most closely related to Paenibacillus yonginensis DCY84T and Paenibacillus physcomitrellae XBT (98.8 and 97.1â% sequence similarity). The DNA G+C content of the novel strain was 53.1±0.3 mol%. Strain S8T contained diphosphatidylglycerol, phosphatidylglycerol, two phospholipids, four aminophospholipids, an aminolipid and three unidentified lipids. The major fatty acid was anteiso-branched C15â:â0. The quinone was menaquinone MK-7. The peptidoglycan of strain S8T contained meso-diaminopimelic acid. The DNA-DNA hybridization values of strain S8T with P. yonginensis KCTC 33428T and P. physcomitrellae DSM 29851T were 44â% and 32â%, respectively. Data from the DNA-DNA hybridization, biochemical, phylogenetic and physiological analyses indicate that strain S8T (=KCTC 33848T=JCM 31672T) represents a novel species of the genus Paenibacillus, for which the name Paenibacillus mobilis sp. nov. is proposed.
Assuntos
Paenibacillus/classificação , Filogenia , Microbiologia do Solo , Técnicas de Tipagem Bacteriana , Composição de Bases , Parede Celular/química , DNA Bacteriano/genética , Ácido Diaminopimélico/química , Ácidos Graxos/química , Hibridização de Ácido Nucleico , Paenibacillus/genética , Paenibacillus/isolamento & purificação , Peptidoglicano/química , Fosfolipídeos/química , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Vitamina K 2/análogos & derivados , Vitamina K 2/químicaRESUMO
Viral factors interact with host cellular proteins, leading to dysregulation of signaling pathways. The Wnt pathway is known to participate in embryonic development and oncogenesis under dysregulation conditions. A downstream factor of the Wnt signaling pathway, ß-catenin, activates T-cell factor (TCF)-dependent transcription, which contributes to cell proliferation and tumorigenesis. In this study, we demonstrated that viral protein kinase (vPK) encoded by Kaposi's sarcoma-associated herpesvirus inhibits the Wnt signaling pathway without affecting nuclear localization and expression of ß-catenin. Coimmunoprecipitation and chromatin immunoprecipitation assays revealed that vPK interacts with ß-catenin, reducing the binding affinity on TCF binding regions as well as interactions of ß-catenin with TCF4. Overexpression of vPK led to reduced mRNA expression of cyclin D1, a well-known transcriptional product of Wnt signaling, suggesting that vPK effectively regulates the host signaling pathway through direct interactions with cellular proteins.
Assuntos
Herpesvirus Humano 8/enzimologia , Proteínas Quinases/metabolismo , Fator de Transcrição 4/metabolismo , Via de Sinalização Wnt/fisiologia , beta Catenina/metabolismo , Células HEK293 , HumanosRESUMO
A novel Gram-stain-negative bacterial strain, designated strain S10T, was isolated from soil collected in a rice field in Goyang, South Korea. Cells of strain S10T were strictly aerobic, motile and rod-shaped. Colonies were round, convex, smooth and white. The strain grew optimally at 37 °C, pH 7.0 and 0â% (w/v) NaCl. Phylogenetic analysis of the 16S rRNA gene sequence of strain S10T revealed that the bacterium belongs to the family Comamonadaceae and is related to members of the genus Hydrogenophaga, with Hydrogenophaga caeni EMB71T being its closest relative (97.9â% sequence similarity). The DNA G+C content of strain S10T was 68.2 mol%. Strain S10T contained phosphatidylethanolamine and diphosphatidylglycerol as the major polar lipids. The major fatty acids were C16â:â0 and summed feature 3 (C16â:â1ω7c and/or iso-C15â:â0 2-OH). The predominant respiratory quinone was ubiquinone Q-8. DNA-DNA hybridization values of strain S10T with Hydrogenophaga caeni KCTC 12613T, Hydrogenophaga atypica DSM 15342T and Hydrogenophaga defluvii DSM 15341T were 16.1±4.8, 49.0±3.2 and 21.9±8.8â%, respectively. Based on phylogenetic distinctiveness, DNA-DNA hybridization and specific physiological and biochemical characteristics, strain S10T (=KCTC 52520T=JCM 31711T) is classified as a novel species of the genus Hydrogenophaga, for which the name Hydrogenophagasoli sp. nov. is proposed.
Assuntos
Comamonadaceae/classificação , Oryza , Filogenia , Microbiologia do Solo , Técnicas de Tipagem Bacteriana , Composição de Bases , Comamonadaceae/genética , Comamonadaceae/isolamento & purificação , DNA Bacteriano/genética , Ácidos Graxos/química , Hibridização de Ácido Nucleico , Fosfolipídeos/química , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Ubiquinona/químicaRESUMO
Prostate cancer (PC) is one of the leading causes of cancer death in men. It commonly develops in older males, but the number of younger men diagnosed with the disease has increased in recent years. Hormone therapies, such as chemical and surgical methods that inhibit androgen synthesis or androgen receptor (AR) activation, have been used for advanced disease. However, castration-resistant PC (CRPC), which exhibits androgen-independent mechanisms for activating AR, develops after a few years of such treatment and no therapy is available. In this study, we examined differences in the proteomes associated with the androgen-dependent (DHT treatment) and -independent (FSK, forskolin treatment) AR signaling conditions in LNCaP prostate cancer cells. Moreover, we used EPI-001, which inhibits AR-mediated transcriptional activity, to examine whether the observed differences in protein expression were directly affected by AR-mediated mechanisms. A total of 213 protein spots were matched in our 2-dimensional gel electrophoresis (2DE) analysis and 8 proteins with significant expression changes in our 5 different treatment groups were identified by mass spectrometry. Among these proteins, expression levels of PEPCK-M, catalase, tubulin alpha chain, alpha-enolase, and endoplasmic reticulum resident protein 29 were significantly altered by DHT and the levels of HSP 90 and EF-Tu were changed by FSK. These changes were blocked by EPI-001 in DHT-regulated proteins, PEPCK-M, catalase, and tubulin alpha chain and FSK-regulated EF-Tu protein. The results from our immunohistochemical analysis using in vivo xenograft models were consistent with the 2DE data. We therefore report the first identification of differences in proteins that are significantly regulated under androgen-dependent and -independent AR signaling conditions. Our findings could suggest a possible molecular mechanism through which AR is activated in the absence and/or presence of androgen, and might help explain the inhibitory action of EPI-001 on CRPC.
Assuntos
Androgênios/farmacologia , Neoplasias da Próstata/metabolismo , Proteoma/metabolismo , Transdução de Sinais/efeitos dos fármacos , Animais , Compostos Benzidrílicos/farmacologia , Linhagem Celular Tumoral , Cloridrinas/farmacologia , Colforsina/farmacologia , Di-Hidrotestosterona/farmacologia , Eletroforese em Gel Bidimensional , Humanos , Imuno-Histoquímica , Masculino , Camundongos , Fator Tu de Elongação de Peptídeos/metabolismo , Fosfoenolpiruvato Carboxiquinase (ATP)/metabolismo , Neoplasias da Próstata/patologia , Proteômica , Espectrometria de Massas em Tandem , Tubulina (Proteína)/metabolismo , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
A yellow-colored Gram-negative strain, Arct 1-12T, was isolated from a soil sample collected in Seoul Women's University, South Korea, and grown on R2A agar at 25 °C. Growth of strain Arct 1-12T was observed at a temperature range of 15-30 °C (optimal 25 °C), but not at 4 or 42 °C. The strain tolerated up to 1% NaCl (w/v) and displayed optimal growth in the absence of NaCl. Growth occurred at pH 6.0-9.0 (optimally at pH 7). According to the 16S rRNA gene sequence, the strain is moderately related to Spirosoma spitsbergense DSM 19989T (93.54%), S. endophyticum EX36T (93.25%), S. linguale LMG 10896T (92%), S. luteum DSM 19990T (93.16%), S. panaciterrae DSM 21099T (91.09%), S. oryzae RHs22T (90.37%), and S. rigui WPCB118T (91.54%). Chemotaxonomic analyses revealed that strain Arct 1-12T possesses MK-7 as the predominant menaquinone, a polar lipid profile consisting of phosphatidylethanolamine, an unknown aminolipid, an unknown aminophospholipid, and an unknown lipid, and iso-C15:0, C16:1 ω5c and Summed Feature 3 (C16:1 ω6c and/or C16:1 ω7c) as the major fatty acids. The DNA G+C content is 52.3 mol %. Based on polyphasic evidence, strain Arct 1-12T (=JCM 31025 T = KCTC 42814T) is classified as the type strain of a novel Spirosoma species for which the name Spirosoma areae sp. nov. is proposed.
Assuntos
Cytophagaceae/classificação , Microbiologia do Solo , Técnicas de Tipagem Bacteriana , Composição de Bases , Cytophagaceae/genética , Cytophagaceae/isolamento & purificação , Cytophagaceae/metabolismo , Código de Barras de DNA Taxonômico , Metabolômica/métodos , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNARESUMO
Strain Mol12T, which presented in the form of Gram-negative, motile, non-spore forming rod-shaped, was isolated from soil in South Korea and characterized to determine its taxonomic position. The strain grew at 20-30°C (optimum 30°C) and pH 7.0-10.0 (optimum pH 8.0) with 1% (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences showed that the strain Mol12T was most closely related to Ensifer terangae LMG 7834T (96.78%), Rhizobium daejeonense KCTC 12121T (96.43%), Ensifer adhaerens Casida AT (96.28%). Chemotaxonomic data showed that the predominant fatty acids were Summed Feature 8 (C18:1 ω7c and/or C18:1 ω6c; 53.02%) and C18:1 ω7c 11-methyl (24.01%). Its complex polar lipid contained major amounts of diphosphatidylglycerol (DPG), phosphatidylethanolamine (PE) and Q-10 as the predominant ubiquinone. The DNA G+C content of strain Mol12T was determined to be 60.9 mol%. Based on the phylogenetic, chemotaxonomic, and phenotypic data, strain Mol12T (=KCTC 42816T =JCM 31049T) ought to be classified as a type strain of a novel species, for which the name Ensifer collicola sp. nov. is proposed.
Assuntos
Rhizobiaceae/isolamento & purificação , Microbiologia do Solo , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano , DNA Ribossômico , Ácidos Graxos/análise , Hibridização de Ácido Nucleico , Fosfolipídeos/análise , Filogenia , RNA Ribossômico 16S/genética , República da Coreia , Rhizobiaceae/classificação , Rhizobiaceae/genética , Rhizobiaceae/fisiologia , Rhizobium , Análise de Sequência de DNARESUMO
Asian dust or yellow sand events in East Asia are a major issue of environmental contamination and human health, causing increasing concern. A high amount of dust particles, especially called as particulate matter 10 (PM10), is transported by the wind from the arid and semi-arid tracks to the Korean peninsula, bringing a bacterial population that alters the terrestrial and atmospheric microbial communities. In this study, we aimed to explore the bacterial populations of Asian dust samples collected during November-December 2014. The dust samples were collected using the impinger method, and the hypervariable regions of the 16S rRNA gene were amplified using PCR followed by pyrosequencing. Analysis of the sequencing data were performed using Mothur software. The data showed that the number of operational taxonomic units and diversity index during Asian dust events were higher than those during non-Asian dust events. At the phylum level, the proportions of Proteobacteria, Actinobacteria, and Firmicutes were different between Asian dust and non-Asian dust samples. At the genus level, the proportions of the genus Bacillus (6.9%), Arthrobacter (3.6%), Blastocatella (2%), Planomicrobium (1.4%) were increased during Asian dust compared to those in non-Asian dust samples. This study showed that the significant relationship between bacterial populations of Asian dust samples and non-Asian dust samples in Korea, which could significantly affect the microbial population in the environment.
Assuntos
Microbiologia do Ar , Poluentes Atmosféricos/análise , Poeira/análise , Material Particulado/análise , Monitoramento Ambiental/métodos , Humanos , Metagenômica , RNA Ribossômico 16S/genética , SeulRESUMO
A yellow-pigmented and strictly aerobic bacterial strain, designated FJY8T, was isolated from the soil of Goyang, South Korea. The cells of FJY8T were Gram-reaction-negative, non-motile rods. Colonies were circular, convex and transparent. Strain FJY8T grew optimally at 30 °C, with 0â% (w/v) NaCl and at pH 8. Phylogenetic analysis of the 16S rRNA gene sequence of FJY8T revealed a clear affiliation of this bacterium to the family Lysobacteraceae, and it was related to members of the genus Lysobacter, with Lysobacter xinjiangensis KCTC 22558T being its closest relative (98.7â% sequence similarity). The DNA G+C content was 68.0±0.4 mol%. Diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol were identified as the major polar lipids, and an unidentified phospholipid and two unidentified aminophospholipids were also detected as the minor polar lipids. The major fatty acids were iso-C16â:â0, summed feature 9 (iso-C17â:â1ω9c and/or C16â:â0 10-methyl) and iso-C15â:â0. Only ubiquinone-8 (Q-8) was detected as the isoprenoid quinone. DNA-DNA hybridization values of strain FJY8T with Lysobacter xinjiangensisRCML-52T and Lysobacter mobilis9NM-14T were 55.8±2.0 and 45.2±4.8â%, respectively. On the basis of DNA-DNA hybridization, phylogenetic distinctiveness, and some physiological and biochemical tests, strain FJY8T (=KCTC 42810T=JCM 31019T) represents a novel species of the genus Lysobacter, for which the name Lysobacter humi sp. nov. is proposed.
Assuntos
Lysobacter/classificação , Filogenia , Microbiologia do Solo , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Lysobacter/genética , Lysobacter/isolamento & purificação , Hibridização de Ácido Nucleico , Fosfolipídeos/química , Pigmentação , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Ubiquinona/químicaRESUMO
Kaposi's sarcoma-associated herpesvirus (KSHV) is associated with formation of Kaposi's sarcoma, multicentric Castleman's disease, and primary effusion lymphoma. Replication and transcription activator (RTA) genes are expressed upon reactivation of KSHV, which displays a biphasic life cycle consisting of latent and lytic replication phases. RTA protein expression results in KSHV genome amplification and successive viral lytic gene expression. Transcriptional activity of viral lytic genes is regulated through epigenetic modifications. In Raji cells latently infected with Epstein-Barr virus, various modifications, such as acetylation and methylation, have been identified at specific lysine residues in histone H4 during viral reactivation, supporting the theory that expression of specific lytic genes is controlled by histone modification processes. Data obtained from chromatin immunoprecipitation and quantitative real-time PCR analyses revealed alterations in the H4K8ac and H4K20me3 levels at lytic gene promoters during reactivation. Our results indicate that H4K20me3 is associated with the maintenance of latency, while H4K8ac contributes to KSHV reactivation in infected TREx BCBL-1 RTA cells.
RESUMO
During Asian dust events, a relatively high concentration of particulate matter is transported by wind from arid and semi-arid regions, such as the Gobi and Taklamakan deserts, to nearby countries, including China, Korea, and Japan. The dust particles contain various microorganisms, which can affect human health as well as the environmental microbe population. In the current study, we investigated the characteristics of the airborne bacterial community during Asian dust events between February and March 2015 in South Korea. Bacterial diversity indexes such as operational taxonomic units, Chao1 and Inverse Simpson index were increased, along with total 16S rRNA gene copy number during Asian dust events. The bacterial community structure during Asian dust events was clearly distinguishable from that during non-Asian dust days. The genera Bacillus and Modestobacter were increased 3.9- and 2.7-fold, respectively, while Escherichia-Shigella was decreased by 89.8%. A non-metric multidimensional scaling plot with metadata analysis revealed association of particulate matter concentration, but not temperature, humidity or wind speed, with bacterial community structure, suggesting that the newly transported dust particles contain various microorganisms that influence the airborne bacterial environment.
Assuntos
Microbiologia do Ar , Poluentes Atmosféricos/análise , Poeira/análise , Material Particulado/análise , Vento , Bactérias/classificação , Bactérias/genética , China , Monitoramento Ambiental/métodos , Humanos , Japão , Filogenia , RNA Ribossômico 16S/genética , República da Coreia , Estações do Ano , Análise de Sequência de DNARESUMO
Kaposi's sarcoma-associated herpesvirus (KSHV) is an etiological agent of Kaposi's sarcoma and primary effusion lymphoma. Like other herpesviruses, KSHV has two distinct life cycles: latent and lytic. Among KSHV latent genes, viral interferon regulatory factor 3 (vIRF3), which shares homology with cellular IRFs, is a multifunctional protein. To identify unknown functions of vIRF3, we performed luciferase-reporter assays in the presence of vIRF3. These analyses revealed that overexpression of vIRF3 inhibited T-cell factor (TCF)-dependent transcriptional activity. This TCF-dependent transcription was associated with the Wnt signaling pathway, which normally regulates embryonic development, but contributes to oncogenesis under dysregulated conditions. Using a mutagenesis analysis, we identified a CREB-binding protein-interaction motif (LXXLL) in vIRF3 as an important region for its inhibitory activity. Collectively, our findings provide insight into the dysregulation of host signaling pathways in KSHV-infected cells.
Assuntos
Proteína de Ligação a CREB/química , Herpesvirus Humano 8/fisiologia , Interações Hospedeiro-Patógeno/genética , Fatores Reguladores de Interferon/metabolismo , Fatores de Transcrição TCF/antagonistas & inibidores , Proteínas Virais/metabolismo , Motivos de Aminoácidos , Núcleo Celular/metabolismo , Células HEK293 , Herpesvirus Humano 8/metabolismo , Humanos , Fatores Reguladores de Interferon/química , Fatores Reguladores de Interferon/genética , Mutação , Fatores de Transcrição TCF/metabolismo , Transcrição Gênica , Proteínas Virais/química , Proteínas Virais/genética , Latência Viral , Via de Sinalização Wnt , beta Catenina/metabolismoRESUMO
A gram-positive, nonmotile, rod-shaped, nonflagellated, aerobic bacterium, designated strain DSD2(T) was isolated from a seawater sample from Sadong wharf, Ulleung-Island, South Korea. Strain DSD2(T) was found to be able to grow at pH ranging from 6 to 11 (optimum 7-8), 0-7 % (w/v) NaCl (optimum 0 %), at 10-42 °C (optimum 37 °C). 16S rRNA gene sequence analysis revealed that strain DSD2(T) was 95.8 % similar to the type strain of Kineosporia rhamnosa KACC 15195(T), 95.8 % similar to Angustibacter aerolatus KACC 15527(T), and 95.5 % similar to Kineococcus xinjiangensis KCTC 19474(T) as its closest relatives. A neighbor-joining phylogenetic tree based on the 16S rRNA gene sequence showed that strain DSD2(T) related to Micrococcineae and Kineosporiineae suborder clade. The major polar lipids were phosphoglycolipids and phospholipids. Strain DSD2(T) was found to contain MK-8 (H2) and MK-9 (H4) as the predominant menaquinone and iso-C16:0 as the major fatty acid. The isolate contained meso-diaminopimelic acid (meso-A2pm) with alanine, glutamic acid, and glycine as the diagnostic diamino acid. The DNA G+C content of strain DSD2(T) was 73.2 mol%. On the basis of phylogenetic, biochemical, chemotaxonomic, and other physiological characteristics, strain DSD2(T) is assigned to a novel species of a novel genus within the suborder Kineosporiineae and the name Thalassiella azotovora gen nov., sp. nov., is proposed. The type strain is DSD2(T) (= KCTC 39634(T) = JCM 31134(T)).