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This study aimed to determine changes in the hydrolysis of vicagrel, a substrate drug of arylacetamide deacetylase (Aadac) and carboxylesterase 2 (Ces2), in P-glycoprotein (P-gp)-deficient or P-gp-inhibited mice and to elucidate the mechanisms involved.Male wild-type (WT) and P-gp knock-out (KO) mice were used to investigate the systemic exposure of vicagrel thiol active metabolite H4 and platelet response to vicagrel, and the mRNA and protein expression levels of intestinal Aadac and Ces2. Moreover, WT mice were administered vicagrel alone or in combination with elacridar (a potent P-gp inhibitor) to determine drug-drug interactions.Compared with WT mice, P-gp KO mice exhibited significant increases in the systemic exposure of H4, the protein expression levels of intestinal Aadac and Ces2, and inhibition of ADP-induced platelet aggregation by vicagrel. Further, the H4 exposure was positively correlated with intestinal Aadac protein expression levels but did not vary with short-term inhibition of P-gp efflux activity by elacridar.P-gp-deficient mice, rather than elacridar-treated mice, exhibited significant upregulation of intestinal Aadac and Ces2 and thus, enhanced metabolic activation of and platelet response to vicagrel, suggesting that the metabolic activation of vicagrel may vary with P-gp deficiency, not P-gp inhibition, in mice.
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Oomycete pathogens such as Phytophthora secrete a repertoire of effectors into host cells to manipulate host immunity and benefit infection. In this study, we found that an RxLR effector, Avr1d, promoted Phytophthora sojae infection in soybean hairy roots. Using a yeast two-hybrid screen, we identified the soybean E3 ubiquitin ligase GmPUB13 as a host target for Avr1d. By coimmunoprecipitation (Co-IP), gel infiltration, and isothermal titration calorimetry (ITC) assays, we confirmed that Avr1d interacts with GmPUB13 both in vivo and in vitro. Furthermore, we found that Avr1d inhibits the E3 ligase activity of GmPUB13. The crystal structure Avr1d in complex with GmPUB13 was solved and revealed that Avr1d occupies the binding site for E2 ubiquitin conjugating enzyme on GmPUB13. In line with this, Avr1d competed with E2 ubiquitin conjugating enzymes for GmPUB13 binding in vitro, thereby decreasing the E3 ligase activity of GmPUB13. Meanwhile, we found that inactivation of the ubiquitin ligase activity of GmPUB13 stabilized GmPUB13 by blocking GmPUB13 degradation. Silencing of GmPUB13 in soybean hairy roots decreased P. sojae infection, suggesting that GmPUB13 acts as a susceptibility factor. Altogether, this study highlights a virulence mechanism of Phytophthora effectors, by which Avr1d competes with E2 for GmPUB13 binding to repress the GmPUB13 E3 ligase activity and thereby stabilizing the susceptibility factor GmPUB13 to facilitate Phytophthora infection. This study unravels the structural basis for modulation of host targets by Phytophthora effectors and will be instrumental for boosting plant resistance breeding.
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Complexos Multiproteicos/química , Phytophthora/química , Ubiquitina-Proteína Ligases/química , Complexos Multiproteicos/metabolismo , Phytophthora/metabolismo , Doenças das Plantas/microbiologia , Ligação Proteica , Ubiquitina-Proteína Ligases/metabolismoRESUMO
To address the lightweight and real-time issues of coal sorting detection, an intelligent detection method for coal and gangue, Our-v8, was proposed based on improved YOLOv8. Images of coal and gangue with different densities under two diverse lighting environments were collected. Then the Laplacian image enhancement algorithm was proposed to improve the training data quality, sharpening contours and boosting feature extraction; the CBAM attention mechanism was introduced to prioritize crucial features, enhancing more accurate feature extraction ability; and the EIOU loss function was added to refine box regression, further improving detection accuracy. The experimental results showed that Our-v8 for detecting coal and gangue in a halogen lamp lighting environment achieved excellent performance with a mean average precision (mAP) of 99.5%, was lightweight with FLOPs of 29.7, Param of 12.8, and a size of only 22.1 MB. Additionally, Our-v8 can provide accurate location information for coal and gangue, making it ideal for real-time coal sorting applications.
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Severe aplastic anemia (SAA) is a bone marrow failure disorder caused by autoimmune dysfunction. The presentation by dendritic cells (DCs) is the key step in initiating the immune response against unknown antigens in SAA patients. In the previous phase, we found that compared to healthy controls, patients with SAA had an increased proportion of circulating myeloid/conventional dendritic cells (mDCs/cDCs) with enhanced phagocytosis, more secretion of Th1-type cytokines (IL-2, TNF-α, IFN-γ) in the bone marrow, and a reduced proportion of Treg cells. In this study, we found that cDCs sorted from SAA patients had higher expression level of HLA-DQ, co-stimulatory molecules CD86, PTK and ERK1/2 than the remission SAA patients and healthy controls. Moreover, downregulation of HLA-DQ protein levels on cDCs derived from SAA patients resulted in reduced phagocytosis rate and CD86 expression of cDCs. When the cDCs above were co-cultured with CD4+ cells from the same patients, reduced secretion of Th1 type of lymphocyte cytokines was observed. Analysis of clinically relevant data suggests that HLA-DQ expression levels were closely related to disease severity and immune status of patients. These findings show that the role of HLA-DQ in the immunopathogenesis of SAA is potentially important and worth further study.
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Anemia Aplástica , Humanos , Medula Óssea/patologia , Fator de Necrose Tumoral alfa , Antígenos HLA-DQ/metabolismoRESUMO
It is meaningful to understand the conversion pathways of nitrogen during the hydrothermal liquefaction process of microalgae to reveal the related reaction mechanisms and develop effective methods to prevent N from ending in biocrude, which eventually increases the quality of biocrude. Extending from our previous works that mainly focused on two high-protein (>50 wt%) microalgae (Chlorella sp. and Spirulina sp.), Nannochloropsis sp., which has a high lipid content (>70 wt%), was used as the feedstock for this project using the same methodology. The high lipid content in Na. induced less nitrogen during the oil phase and as a result, reduced the heteroatom content while also improving the quality of biocrude. It is worth noting that another investigation was conducted on the model compounds with different types of amino acids to specify the effects of the types of amino acids in the proteins in microalgae on the N pathway and their distribution in the products (aqueous phase, oil, solid, and gas). It was found that the basic amino acid in microalgae caused the formation of more N-heterocyclic compounds in the biocrude. The mass flow based on the mass balance was demonstrated to further refine the map showing the predicted reaction pathway of nitrogen from the previous version.
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Chlorella , Microalgas , Temperatura , Aminoácidos , Microalgas/química , Nitrogênio , Água , Lipídeos , Biocombustíveis , BiomassaRESUMO
Sludge retention time (SRT) regulation is one of the essential management techniques for refined control of the main-sidestream treatment process under the low ammonia density. It is indispensable to understand the effect of SRTs changes on the Nitrifier kinetics to obtain the functional separation of the Nitrifier and the refined control of the nitrification process. In this study, Nitrifier was cultured with conditions of 35 ± 0.5 °C, pH 7.5 ± 0.2, DO 5.0 ± 0.5 mg-O/L, and SRTs were controlled for 40 d, 20 d, 10 d, and 5 d. The net growth rate (µm), decay rate (b), specific growth rate (µ), the yield of the Nitrifier (YA), temperature parameter (TA), and inhibition coefficient (KI) have been measured and extended with the SRT decreases. Instead, the half-saturation coefficient (KS) decreased. In addition, the limited value of pH inhibition occurs (pHUL), and the pH of keeping 5% maximum reaction rate (pHLL) was in a relatively stable state. The trade of kinetics may be induced by the change of species structure of Nitrifier. The Nitrosomonas proportion was increased, and the Nitrospira was contrary with the SRT decreasing. It is a match for the functional separation of Nitrifier when SRTs was 20 d at ambient temperature under the low ammonia density. The kinetics of ammonia-oxidizing organisms (AOO) and nitrite-oxidizing organisms (NOO) in Nitrifier under different SRT conditions should be measured respectively to the refined control of the partial nitrification process in future study.
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Amônia , Esgotos , Reatores Biológicos , Cinética , TemperaturaRESUMO
This study investigated the expression status of signaling lymphocytic activation molecule family 6 (SLAMF6) in CD8+ T lymphocytes of patients with severe aplastic anemia (SAA) and its association with the clinical indicators and immune status of the disease. The effects of SLAMF6 on the function and apoptosis of CD8+ T lymphocytes were also investigated. Levels of SLAMF6 and SLAM-associated protein in the CD8+ T lymphocytes of SAA patients were significantly lower than the normal controls, and they were positively correlated with hematopoietic-related indicators but negatively correlated with the levels of functional molecules of CD8+ T lymphocytes. After blocking SLAMF6, CD8+ T lymphocyte functional molecule secretion was upregulated and RICD was downregulated in SAA patients, suggesting that SLAMF6, is involved in the pathogenetic mechanism of SAA by regulating CD8+ T lymphocyte functional molecule secretion and RICD levels. SLAMF6 may be a novel target for the regulation of CD8+ T lymphocyte homeostasis.
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Anemia Aplástica/metabolismo , Linfócitos T CD8-Positivos/imunologia , Família de Moléculas de Sinalização da Ativação Linfocitária/metabolismo , Adulto , Idoso , Anemia Aplástica/imunologia , Apoptose , Citotoxicidade Imunológica , Regulação para Baixo , Feminino , Hematopoese , Humanos , Masculino , Pessoa de Meia-Idade , Índice de Gravidade de Doença , Família de Moléculas de Sinalização da Ativação Linfocitária/genética , Adulto JovemRESUMO
Exosomes are membrane-bound extracellular vesicles that are produced in the endosomal compartment of most eukaryotic cells. Containing proteins, RNA, and DNA, exosomes mediate intercellular communication between different cell types by transferring their contents and thus are involved in numerous physiological and pathological processes. T cells are an indispensable part of adaptive immunity, and the functions of T cell-derived exosomes have been widely studied. In the more than three decades since the discovery of exosomes, several studies have revealed that T cell-derived exosomes play a novel role in cell-to-cell signaling, especially in inflammatory responses, autoimmunity, and infectious diseases. In this review, we will summarize the function of T cell-derived exosomes and their therapeutic potential.
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Exossomos/fisiologia , Linfócitos T/ultraestrutura , Linfócitos T CD4-Positivos/ultraestrutura , Linfócitos T CD8-Positivos/ultraestrutura , Humanos , Linfócitos T Reguladores/ultraestruturaRESUMO
Applying silver into coatings has become a prevalent method in fabricating antimicrobial surfaces. However, the concerns about durability always exist and limit its applications. Here, a highly inhibitory, active, durable, and easy-to-use silver ions-nanosilver antimicrobial additive for powder coatings was fabricated in this study. Silver nanoparticles were chemically bonded to the Ag, Cu, and Zn-ternary ion-exchanged zeolite by α -lipoic acid, which was then encapsulated by hydrophilic polymers. The fabricated silver ions and silver nanoparticles (Ag+-AgNPs) complementary structure provides a synergistic effect. Ag+ is the main antimicrobial agent, while AgNPs act as a supplementary reservoir of Ag+. As well, the formed thin layer of silver nanoparticles and hydrophilic film prolongs the release of active Ag+ from zeolite, and Ag+ facilitates the activation of AgNPs. The results show that this additive indicates excellent antimicrobial activity to E. coli, S. aureus, P. aeruginosa, and C. albicans, and that the coatings with the additive exhibit over 99.99% reduction rate for the tested bacteria and fungi. The coating film is able to maintain over 99% antimicrobial reduction even after 1200 repeated solution wipings, or over 30 wash cycles of artificial sweat solution, indicating high durability. Furthermore, the yellowness of the coating is not evident (Δb < 2) despite the high loading of silver, and the silver nanoparticles have little impact on gloss, haze, and distinctness of the coating film image.
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BACKGROUND: Glucagon-like peptide 1 (GLP-1) and its analogs are first-line choices for the treatment of type 2 diabetes mellitus. Recent studies have shown that they exhibit antitumor properties in some tumors. We previously found that a GLP-1 analog, exendin-4 (Ex-4), inhibited the growth of prostate cancer cells through suppressing the PI3K/Akt/mTOR pathway, which is activated in response to enzalutamide treatment and reported to be closely related to resistance to enzalutamide. So we speculated that exendin-4 may enhance the sensitivity of prostate cancer to enzalutamide through inhibiting Akt activation. METHODS: LNCap and CWR22RV1 cell lines, as well as mice bearing xenografts formed from the two cells, were used. RESULTS: Exendin-4 in combination with enzalutamide dramatically suppressed tumor growth of prostate cancer cells compared to enzalutamide alone; exendin-4 is capable of antagonizing enzalutamide-induced invasion and migration of both prostate cancer cells (P < .05). Furthermore, the combination treatment significantly reduced Akt and mTOR levels that were triggered by enzalutamide administration, caused a further decrease in nuclear AR localization compared with the enzalutamide as a monotherapy (P < .5), though exendin-4 treatment alone showed no effect on nuclear AR. CONCLUSION: Our study demonstrated that exendin-4 alleviated resistance to enzalutamide, and suggested that exendin-4 combined with enzalutamide may be a more efficacious treatment for patients with advanced prostate cancer.
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Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Exenatida/farmacologia , Feniltioidantoína/análogos & derivados , Neoplasias da Próstata/tratamento farmacológico , Proteínas Proto-Oncogênicas c-akt/metabolismo , Animais , Benzamidas , Processos de Crescimento Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Núcleo Celular/metabolismo , Sinergismo Farmacológico , Ativação Enzimática/efeitos dos fármacos , Exenatida/administração & dosagem , Receptor do Peptídeo Semelhante ao Glucagon 1/biossíntese , Humanos , Masculino , Camundongos Endogâmicos BALB C , Camundongos Nus , Invasividade Neoplásica , Nitrilas , Feniltioidantoína/administração & dosagem , Feniltioidantoína/farmacologia , Neoplasias da Próstata/enzimologia , Neoplasias da Próstata/patologia , Distribuição Aleatória , Receptores Androgênicos/metabolismo , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
This study is aimed at investigating the effects of shikonin, a pyruvate kinase M2 (PKM2) inhibitor, on the functions of myeloid dendritic cells (mDCs) in a mouse model of severe aplastic anemia (AA) generated by total body irradiation and lymphocyte infusion. Flow cytometry and qPCR were used to determine the proportions of PKM2+ mDCs and other immune indicators in the AA mice. Glucose consumption level, pyruvate generation level, and ATP content were used to determine the level of glycolytic metabolism in the mDCs. The survival rates of AA mice were evaluated after the administration of shikonin or the immunosuppressive agent cyclosporin A. The AA mice displayed pancytopenia, decreased CD4+/CD8+ cell ratio, increased perforin and granzyme levels in CD8+ cells, increased costimulatory CD80 and CD86 expressions, and inadequate regulatory T cell number. In vivo animal experiments showed that the shikonin-mediated inhibition of the PKM2 expression in mice was associated with high survival rates. In addition, the administration of cyclosporin A or shikonin decreased the expression of cytotoxic molecules and costimulatory CD80 and CD86 on CD8+ cells. Taken together, the results of this study indicated that shikonin could inhibit the activation and proliferation of mDCs as well as the activation of downstream cytotoxic T cells by reducing the PKM2 level in mDCs.
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Anemia Aplástica/tratamento farmacológico , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/metabolismo , Naftoquinonas/uso terapêutico , Anemia Aplástica/imunologia , Anemia Aplástica/metabolismo , Animais , Antígeno B7-1/metabolismo , Antígeno B7-2/metabolismo , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/metabolismo , Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/metabolismo , Ciclosporina/uso terapêutico , Citometria de Fluxo , Imunossupressores/uso terapêutico , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Piruvato Quinase/genética , Piruvato Quinase/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
BACKGROUND: Regulatory T cells (Tregs) inhibit the activation of cluster of differentiation (CD) 4+ , CD8+ T cells and the antigen-presenting process of antigen-presenting cells, and may play an important role in acquired severe aplastic anemia (SAA). METHODS: Flow cytometry was used to measure CD4+ CD25+ CD127dim Tregs, cytotoxic T lymphocyte antigen 4 (CTLA-4) expression on Tregs, and human leukocyte antigen (HLA)-DQ expression on myeloid dendritic cells (mDCs). The correlations of CTLA-4 and HLA-DQ with immune status and clinical indicators and the changes in these indicators after immunosuppressive therapy (IST) were analyzed. RESULTS: In SAA patients, the number of Tregs and their CTLA-4 expression were low but recovered after IST; the HLA-DQ expression on mDCs was high but decreased after IST. The CTLA-4 expression on Tregs and the HLA-DQ expression on mDCs showed a negative correlation. The CTLA-4 on Tregs was positively but HLA-DQ on mDCs negatively correlated with the number of Tregs, natural killer (NK) cell number, and CD4+ T/CD8+ T ratio. CTLA-4 was positively but HLA-DQ negatively correlated with the percentage of granulocytoid and erythroid cells in bone marrow, white blood cell count in PB, absolute neutrophil count in PB, and the percentage of reticulocytes in PB. CONCLUSIONS: CTLA-4/HLA-DQ may be key in the regulation of Tregs on mDCs in SAA patients. Our findings should be helpful for further investigation of the mechanism of immune pathogenesis in SAA patients. Studies on the regulators of Treg and CTLA-4 activity will be valuable for SAA therapeutic target research and disease monitoring.
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Anemia Aplástica/imunologia , Antígeno CTLA-4/metabolismo , Células Dendríticas/imunologia , Antígenos HLA-DQ/metabolismo , Imunidade Celular , Linfócitos T Reguladores/imunologia , Adolescente , Adulto , Idoso , Criança , Feminino , Humanos , Células Matadoras Naturais/imunologia , Contagem de Linfócitos , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
The objective of this study is to explore the optimal pre-treatment procedures and statistics methods for live/dead bacterial staining using nitrite oxidizing organism (NOO) as the research aim. This staining method was developed and widely utilized to evaluate activated bacterial survival situation, because it is direct and convenience to count live and dead bacteria amount by colour distinguishes (green/red) from pictures taken by microscope. The living cell (green colour) percentage and initial bacterial chemical oxygen demand (COD) could be used for accurate reaction rate calculation at the beginning of tests. While according to the physiological principles, the detection target was limited as the organism has a complete cell shape, that was applicable for the initial phase for decay stage (live cell â particulate dead cell), but it is impossible to evaluate the decayed soluble COD from particulate dead cell during whole reaction. To model the decay stage scientifically, a two-step decay model was developed to cater to the live/dead bacterial staining analysis of biological nitrite oxidizer under inhibition condition of high nitrite concentrations at 35 °C. As results of optimal pre-treatment, a three level ultrasonic wave with 45 seconds was explored, as a reasonable observed picture number, 30 sets with 95% confident interval for datasets statistics was summarized. A set of nitrite oxidizer inhibition test (total COD and oxygen uptake rates) under high nitrite concentrations was simulated using the above model and obtained experimental schemes. Additionally, the disintegration enhancement from particulate dead cell to soluble COD by nitrite was inspected and modelled on the basis of experimental datasets.
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Nitritos , Esgotos , Bactérias , Análise da Demanda Biológica de Oxigênio , Reatores Biológicos , OxirreduçãoRESUMO
We identified a glycoside hydrolase family 12 (GH12) protein, XEG1, produced by the soybean pathogen Phytophthora sojae that exhibits xyloglucanase and ß-glucanase activity. It acts as an important virulence factor during P. sojae infection but also acts as a pathogen-associated molecular pattern (PAMP) in soybean (Glycine max) and solanaceous species, where it can trigger defense responses including cell death. GH12 proteins occur widely across microbial taxa, and many of these GH12 proteins induce cell death in Nicotiana benthamiana. The PAMP activity of XEG1 is independent of its xyloglucanase activity. XEG1 can induce plant defense responses in a BAK1-dependent manner. The perception of XEG1 occurs independently of the perception of ethylene-inducing xylanase. XEG1 is strongly induced in P. sojae within 30 min of infection of soybean and then slowly declines. Both silencing and overexpression of XEG1 in P. sojae severely reduced virulence. Many P. sojae RXLR effectors could suppress defense responses induced by XEG1, including several that are expressed within 30 min of infection. Therefore, our data suggest that PsXEG1 contributes to P. sojae virulence, but soybean recognizes PsXEG1 to induce immune responses, which in turn can be suppressed by RXLR effectors. XEG1 thus represents an apoplastic effector that is recognized via the plant's PAMP recognition machinery.
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Glycine max/imunologia , Glycine max/microbiologia , Glicosídeo Hidrolases/metabolismo , Moléculas com Motivos Associados a Patógenos/metabolismo , Phytophthora/enzimologia , Phytophthora/patogenicidade , Fatores de Virulência/metabolismo , Bactérias/enzimologia , Capsicum/metabolismo , Morte Celular , Resistência à Doença , Glicosídeo Hidrolases/isolamento & purificação , Hidrólise , Solanum lycopersicum/metabolismo , Mutação/genética , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Imunidade Vegetal , Proteínas de Plantas/metabolismo , Sinais Direcionadores de Proteínas , Glycine max/citologia , Nicotiana/citologia , Nicotiana/metabolismo , Nicotiana/microbiologia , VirulênciaRESUMO
A novel thermophilic actinomycete, designated strain 3-12XT, was isolated from mushroom compost in Guangxi University, Nanning, China. The novel isolate contained meso-diaminopimelic acid as the diagnostic diamino acid and the whole-cell sugars were glucose and ribose. The predominant menaquinones were MK-9(H4) and MK-9(H6). The polar phospholipids were diphosphatidylglycerol, hydroxy-phosphatidylethanolamine, phosphatidylmethylethanolamine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannoside, ninhydrin-positive phosphoglycolipids and glycolipids. Major fatty acids were so-C16â:â0 and C17â:â0. The G+C content of the genomic DNA was 74.6â%. The 16S rRNA gene sequence analysis showed that the closest phylogenetic neighbour of strain 3-12XT was Thermomonospora chromogena ATCC 43196T (97.0â%), other closely related strains all belonged to the family Streptosporangiaceae and showed more than 6â% divergence. The chemotaxonomic characteristics of strain 3-12XT were significantly different from Thermomonospora chromogena ATCC 43196T and DNA-DNA hybridization showed low relatedness (48.6-55.6â%) between them, so they should be different species. Thermomonospora chromogena was removed from the genus Thermomonospora by Zhang et al. 1998 on the basis of phylogenetic, chemotaxonomic and phenotypic evidence, but its taxonomic position remains uncertain. Based on the phenotypic and phylogenetic data, strain 3-12XT represents a novel species in a new genus in the family Streptosporangiaceae. The name Thermostaphylospora griseoalba gen. nov., sp. nov. is proposed. The type strain of Thermostaphylospora grisealba is 3-12XT (=DSM 46781T=CGMCC 4.7160T). We also propose transferring Thermomonospora chromogenaZhang et al. 1998 to Thermostaphylospora chromogena comb. nov. (type strain ATCC 43196T=JCM 6244T).
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Actinomycetales/classificação , Agaricales , Filogenia , Microbiologia do Solo , Actinomycetales/genética , Actinomycetales/isolamento & purificação , Técnicas de Tipagem Bacteriana , Composição de Bases , China , Compostagem , DNA Bacteriano/genética , Ácido Diaminopimélico/química , Ácidos Graxos/química , Glicolipídeos/química , Hibridização de Ácido Nucleico , Fosfolipídeos/química , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Vitamina K 2/análogos & derivados , Vitamina K 2/químicaRESUMO
During an investigation exploring potential sources of novel thermophilic species and natural products, a novel thermophilic and alkaliphilic actinomycete with alkaline cellulase producing ability, designated strain 4-2-13T, was isolated from soil of a tropical rainforest in Xishuangbanna, Yunnan province, China. The morphological and chemotaxonomic characteristics of strain 4-2-13T are consistent with those of the members of the genus Streptomyces. The strain forms extensively branched aerial mycelia and substrate mycelia. Spiral spore chains were observed on aerial mycelia; spores were oval to cylindrical, with smooth surfaces. The organism was found to contain LL-diaminopimelic acid as the diagnostic diamino acid in the cell wall peptidoglycan. The whole cell hydrolysates were found to contain glucose and ribose. The cellular fatty acid profile mainly consists of anteiso-C17:0 and iso-C16:0. The menaquinones were identified as MK-9(H8), MK-10(H6) and MK-9(H6). The polar lipids profile were found to consist of diphosphatidylglycerol, phosphatidylmethylethanolamine, a ninhydrin-positive glycophospholipid, phosphatidylinositol, phosphatidylglycerol and unidentified glycolipids. The 16S rRNA gene sequence analysis showed that the organism belongs to the genus Streptomyces and in the 16S rRNA gene tree it formed a distinct phyletic line together with the closely related type strain Streptomyces burgazadensis Z1R7T (95.2% sequence similarity). However, the phenotypic characteristics of strain 4-2-13T are significantly different from those of S. burgazadensis Z1R7T. Based on the phenotypic, chemotaxonomic and phylogenetic characteristics, strain 4-2-13T represents a novel species in the genus Streptomyces, for which the name Streptomyces thermoalkaliphilus sp. nov. is proposed. The type strain is 4-2-13T (= DSM 42159T = CGMCC 4. 7205T).
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Álcalis/metabolismo , Celulase/biossíntese , Floresta Úmida , Microbiologia do Solo , Streptomyces/classificação , Streptomyces/metabolismo , Metabolômica/métodos , Fenótipo , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Streptomyces/isolamento & purificaçãoRESUMO
BACKGROUND: Immuno-related hemocytopenia (IRH) is defined as idiopathic cytopenia of undetermined significance (ICUS) patients with autoantibodies. In our previous studies, we found that IgG1 levels were increased in IRH patients and might cause the destruction of hematopoietic cells. METHODS: In this study, we analyzed IgG subclasses in 30 IRH patients (male:female = 13:17, median age 32 years, range 18 - 56), 15 IRH remission patients (IRH-R) (male:female = 6:9, median age 34, range 20 - 52) and 20 normal controls (male:female = 8:12, median age 27, range 24 - 36) by Cytometric Bead Array, Flow Cytometry and Immunohistochemical staining. RESULTS: Levels of IgG1/IgG3 in the bone marrow supernatant of IRH patents, as well as the proportion of CD5+ B lymphocytes and Th2 cells (CD3+CD8-IL-4+) were higher than those of IRH-R patients and normal controls, and IgG1 levels had a positive correlation with the proportion of Th2 cells. In IRH patients, IgG1 and IgG3 were positive on nucleated erythrocytes and granulocytes, which were negative in IRH-R patients and healthy controls and had inverse correlations with hematopoietic function. Using immunohistochemical staining, IgG1 were also detected on bone marrow biopsies of IRH patients. CONCLUSIONS: The results indicated that IgG1 and IgG3 autoantibodies in IRH patients might play a key role in the IRH pathogenesis and in the abnormal immune function of IRH patients.
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Autoanticorpos/sangue , Imunoglobulina G/sangue , Pancitopenia/sangue , Adolescente , Adulto , Autoanticorpos/imunologia , Autoanticorpos/metabolismo , Criança , Eritrócitos/imunologia , Eritrócitos/metabolismo , Feminino , Citometria de Fluxo , Granulócitos/imunologia , Granulócitos/metabolismo , Humanos , Imunoglobulina G/imunologia , Imunoglobulina G/metabolismo , Linfócitos/imunologia , Linfócitos/metabolismo , Masculino , Pessoa de Meia-Idade , Pancitopenia/imunologia , Pancitopenia/metabolismo , Adulto JovemRESUMO
Severe aplastic anemia (SAA) is an autoimmune disease with destruction of hematopoietic cells by activated T lymphocytes. However, the precise mechanism of cytotoxicity T cells recognizing and attacking CD34(+) cells remains unclear. Here, we investigated the proteome of CD34(+) cells in SAA patients to further explore the pathogenesis of SAA. CD34(+) cells from 29 SAA patients and 20 health controls were isolated by magnetic activated cell sorting. The protein of CD34(+) cells were examined by iTRAQ labeling combination of multidimensional liquid chromatography and tandem mass spectrometry. A total of 156 differential expression proteins in CD34(+) cells were identified. Compared with health controls, 53 proteins were up-regulated and 103 proteins were down-regulated in SAA patients. Specifically, abnormal expression of proteasome subunits, histone variants, dolichyl-diphosphooligosaccharide-protein glycosyltransferase subunit (DAD1) and ATPase inhibitor, mitochondrial isoform 1 precursor(IF1) may relate to the hyperfunction of immune responses and excessive apoptosis of SAA CD34(+) cells.
Assuntos
Anemia Aplástica/metabolismo , Proteínas Reguladoras de Apoptose/metabolismo , Células da Medula Óssea/metabolismo , Proteínas de Membrana/metabolismo , Mitocôndrias/metabolismo , Complexo de Endopeptidases do Proteassoma/metabolismo , Proteínas/metabolismo , Proteoma , Adolescente , Adulto , Antígenos CD34/metabolismo , Apoptose , Células da Medula Óssea/patologia , Criança , Progressão da Doença , Feminino , Histonas/metabolismo , Humanos , Masculino , Pessoa de Meia-Idade , Espectrometria de Massas em Tandem , Adulto Jovem , Proteína Inibidora de ATPaseRESUMO
Heat shock factors (HSFs) are critical regulators of spermatogenesis. However, heat shock responses, the associated components and the underlying functional mechanisms remain to be elucidated. Here, we characterize the expression pattern of HSFY, a member of the HSF family in the testis and epididymis. Its expression in testis and epididymis was initially identified by western blots. Immunofluorescence staining demonstrated that HSFY was confined to the cytoplasm of late spermatocytes and spermatids in adult testes, gonocytes in newborn testes and undifferentiated spermatogonia in 7 days post-parturition testes. In the epididymis, HSFY was predominantly expressed in principal cells. Furthermore, a single transient scrotal heat stress did not change HSFY protein expression in the testes or epididymis, either on the expressional level or in cellular localization. In summary, this study detected the expression pattern of HSFY in the testes and epididymis and demonstrated that its expression was not regulated by transient elevated temperature.