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1.
J Orthop Sci ; 2023 Oct 18.
Artigo em Inglês | MEDLINE | ID: mdl-37863682

RESUMO

BACKGROUND: The purpose of this study was to investigate the clinical effect of an intra-articular and local infiltration injection of a compound analgesic mixture of ropivacaine and compound betamethasone on the repair of the triangular fibrocartilage complex under wrist arthroscopy. METHODS: This prospective, double-blind, randomized study involved 20 patients with Atzei type 2 or 3 injuries of the triangular fibrocartilage complex who underwent repair under wrist arthroscopy. Patients were divided into two groups (n = 10) according to the systematic random sampling method. The test group was injected with a "cocktail" mixture for pain relief. The control group was injected with normal saline. The visual analog scale (VAS) pain score, pinch force, wrist joint mobility, wrist joint function score (PRWE score), occurrence of adverse reactions and dosage of analgesic drugs were evaluated before and after the operation in the two groups. RESULTS: The resting pain of the patients in the test group was less severe than that of the control group at 12 h, 24 h and 48 h after the operation (P < 0.05), and the pinch force of the patients in the test group was significantly greater than that of the control group at 1 d, 2 d and 3 d after the operation (P < 0.01). The amount of postoperative analgesics used in the test group was significantly lower than that in the control group (P < 0.01), and the patient satisfaction rate in the test group was higher than that in the control group (P < 0.05). There were no postoperative adverse effects in either group. CONCLUSION: An intra-articular and local infiltration injection of a "cocktail" analgesic mixture in the repair of triangular fibrocartilage complex under wrist arthroscopy can provide good pain control in the early postoperative period and reduce the amount of postoperative analgesic drugs administered, thus improving clinical safety. LEVEL OF EVIDENCE: Level II; Randomized Controlled Trial; Treatment Study.

2.
J Sci Food Agric ; 103(2): 944-956, 2023 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-36066553

RESUMO

BACKGROUND: Gastric mucosal injury caused by ethanol is a common gastrointestinal disease. Quinoa (Chenopodium quinoa Willd.), as a nutrient-rich grain, plays a significant role in preventing and treating gastric mucosal damage. The present study aimed to explore the protective effect of quinoa on alcohol-induced gastric mucosal damage and its possible mechanism. RESULTS: The ethanol-induced gastric mucosal injury rat model was used for in vivo experiments and H2 O2 -induced GES-1 cells for in vitro experiments to elucidate the protective effect of quinoa. The results show that quinoa water extract can increase the superoxide dismutase level and decrease the malondialdehyde level in vitro and in vivo. Furthermore, quinoa also reduced the bleeding point and bleeding area in rats with ethanol-induced gastric mucosal injury and improved gastric histopathological changes. H2 O2 significantly increased the levels of inflammatory factors in GES-1 cells, which were markedly ameliorated by quinoa water extract. Likewise, quinoa water extract regulated the protein expression levels of Nrf2, Keap1, HO-1, p-IKK, and p-NF-κB through Nrf2 and nuclear factor-κB signaling pathways, reducing the production of oxidative stress and inflammation, thereby repairing the damaged gastric mucosa. CONCLUSION: The findings of this study demonstrated that quinoa shows protective effect against ethanol-induced gastric mucosal injury through its anti-inflammatory and anti-oxidant effects. We propose that our research will provide a reference for quinoa as a functional food. © 2022 Society of Chemical Industry.


Assuntos
Chenopodium quinoa , Úlcera Gástrica , Ratos , Animais , Chenopodium quinoa/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Proteína 1 Associada a ECH Semelhante a Kelch/metabolismo , Mucosa Gástrica/metabolismo , Etanol/metabolismo , Estresse Oxidativo , NF-kappa B/metabolismo , Água/metabolismo , Úlcera Gástrica/induzido quimicamente
3.
Molecules ; 27(20)2022 Oct 20.
Artigo em Inglês | MEDLINE | ID: mdl-36296669

RESUMO

CH4 can be separated from low-concentration coal bed methane (LCCBM) by using the hydrate-based gas separation (HBGS) method. To study the contribution of different cyclic organic compounds to the separation of CH4 in LCCBM, an LCCBM hydrate model was constructed. Based on the Monte Carlo and molecular dynamics theory, we simulated the effect of three cyclic organic compounds-cyclopentane (CP), cyclopentanone (CP-one), and cyclopentanol (CP-ol)-on the stability of the LCCBM hydrate at P = 2 MPa, various temperatures, and discussed the structural stability of the hydrate in depth in terms of final snapshots, radial distribution function, mean square displacement, diffusion coefficient, and potential energy change. The results showed that for the CH4-N2 LCCMM gas mixture, CP showed the best facilitation effect compared to the other two cyclic compounds by maintaining the stability of the LCCBM hydrate well at T = 293 K. The promotion effect of CP-one is between CP and CP-ol, and when the temperature increases to T = 293 K, the oxygen atoms in the water molecule can maintain the essential stability of the hydrate structure, although the orderliness decreases significantly. Moreover, the structure of the hydrate model containing CP-ol is destroyed at T = 293 K, and the eventual escape of CH4 and N2 molecules in solution occurs as bubbles. The research results are important for further exploration of the mechanism of action of cyclic promoter molecules with LCCBM hydrate molecules and promoter preferences.


Assuntos
Dióxido de Carbono , Metano , Dióxido de Carbono/química , Metano/química , Água/química , Simulação de Dinâmica Molecular , Ciclopentanos , Carvão Mineral , Oxigênio
4.
Appl Environ Microbiol ; 85(1)2019 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-30341075

RESUMO

Lincomycin A (Lin-A) is a widely used antibacterial antibiotic fermented by Streptomyces lincolnensis However, the transcriptional regulatory mechanisms underlying lincomycin biosynthesis have seldom been investigated. Here, we first identified a TetR family transcriptional regulator (TFR), SLCG_2919, which negatively modulates lincomycin biosynthesis in S. lincolnensis LCGL. SLCG_2919 was found to specifically bind to promoter regions of the lincomycin biosynthetic gene cluster (lin cluster), including 25 structural genes, three resistance genes, and one regulatory gene, and to inhibit the transcription of these genes, demonstrating a directly regulatory role in lincomycin biosynthesis. Furthermore, we found that SLCG_2919 was not autoregulated, but directly repressed its adjacent gene, SLCG_2920, which encodes an ATP/GTP binding protein whose overexpression increased resistance against lincomycin and Lin-A yields in S. lincolnensis The precise SLCG_2919 binding site within the promoter region of SLCG_2920 was determined by a DNase I footprinting assay and by electrophoretic mobility shift assays (EMSAs) based on base substitution mutagenesis, with the internal 10-nucleotide (nt) AT-rich sequence (AAATTATTTA) shown to be essential for SLCG_2919 binding. Our findings indicate that SLCG_2919 is a negative regulator for controlling lincomycin biosynthesis in S. lincolnensis The present study improves our understanding of molecular regulation for lincomycin biosynthesis.IMPORTANCE TetR family transcriptional regulators (TFRs) are generally found to regulate diverse cellular processes in bacteria, especially antibiotic biosynthesis in Streptomyces species. However, knowledge of their function in lincomycin biosynthesis in S. lincolnensis remains unknown. The present study provides a new insight into the regulation of lincomycin biosynthesis through a TFR, SLCG_2919, that directly modulates lincomycin production and resistance. Intriguingly, SLCG_2919 and its adjoining gene, SLCG_2920, which encodes an ATP/GTP binding protein, were extensively distributed in diverse Streptomyces species. In addition, we revealed a new TFR binding motif, in which SLCG_2919 binds to the promoter region of SLCG_2920, dependent on the intervening AT-rich sequence rather than on the flanking inverted repeats found in the binding sites of other TFRs. These insights into transcriptional regulation of lincomycin biosynthesis by SLCG_2919 will be valuable in paving the way for genetic engineering of regulatory elements in Streptomyces species to improve antibiotic production.


Assuntos
Antibacterianos/biossíntese , Proteínas de Bactérias/genética , Lincomicina/biossíntese , Streptomyces/genética , Fatores de Transcrição/genética , Proteínas de Bactérias/metabolismo , Regulação Bacteriana da Expressão Gênica , Streptomyces/metabolismo , Fatores de Transcrição/metabolismo
5.
J Ind Microbiol Biotechnol ; 46(7): 1013-1024, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31016583

RESUMO

In this work, we found that the Lrp/AsnC family protein SACE_5717 negatively regulated erythromycin biosynthesis in S. erythraea. Disruption of SACE_5717 led to a 27% improvement in the yield of erythromycin in S. erythraea A226. SACE_5717 directly repressed its own gene expression, as well as that of the adjacent gene SACE_5716 by binding to the target sequence 5'-GAACGTTCGCCGTCACGCC-3'. The predicted LysE superfamily protein SACE_5716 directly influenced the export of lysine, histidine, threonine and glycine in S. erythraea. Arginine, tyrosine and tryptophan were characterized as the effectors of SACE_5717 by weakening the binding affinity of SACE_5717. In the industrial S. erythraea WB strain, deletion of SACE_5717 (WBΔSACE_5717) increased erythromycin yield by 20%, and by 36% when SACE_5716 was overexpressed in WBΔSACE_5717 (WBΔSACE_5717/5716). In large-scale 5-L fermentation experiment, erythromycin yield in the engineered strain WBΔSACE_5717/5716 reached 4686 mg/L, a 41% enhancement over 3323 mg/L of the parent WB strain.


Assuntos
Eritromicina/biossíntese , Saccharopolyspora/metabolismo , Engenharia de Proteínas , Saccharopolyspora/genética
6.
Curr Microbiol ; 75(11): 1429-1433, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30030563

RESUMO

Chinese strong-flavor liquor (CSFL), accounting for more than 70% of both Chinese liquor production and sales, was produced by complex fermentation with pit mud. Clostridium kluyveri, an important species coexisted with other microorganisms in fermentation pit mud (FPM), could produce caproic acid, which was subsequently converted to the key CSFL flavor substance ethyl caproate. In this study, we present the first complete genome sequence of C. kluyveri isolated from FPM. Clostridium kluyveri JZZ contains one circular chromosome and one circular plasmid with length of 4,454,353 and 58,581 bp, respectively. 4158 protein-coding genes were predicted and 2792 genes could be assigned with COG categories. It possesses the pathway predicted for biosynthesis of caproic acid with ethanol. Compared to other two C. kluyveri genomes, JZZ consists of longer chromosome with multiple gene rearrangements, and contains more genes involved in defense mechanisms, as well as DNA replication, recombination, and repair. Meanwhile, JZZ contains fewer genes involved in secondary metabolites biosynthesis, transport, and catabolism, including genes encoding Polyketide Synthases/Non-ribosomal Peptide Synthetases. Additionally, JZZ possesses 960 unique genes with relatively aggregating in defense mechanisms and transcription. Our study will be available for further research about C. kluyveri isolated from FPM, and will also facilitate the genetic engineering to increase biofuel production and improve fragrance flavor of CSFL.


Assuntos
Clostridium kluyveri/genética , Genoma Bacteriano , Vinho/microbiologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Sequência de Bases , Caproatos/metabolismo , China , Clostridium kluyveri/isolamento & purificação , Clostridium kluyveri/metabolismo , Etanol/metabolismo , Fermentação , Aromatizantes/metabolismo
7.
J Ind Microbiol Biotechnol ; 45(5): 345-355, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29574602

RESUMO

Streptomyces lincolnensis is generally utilized for the production of lincomycin A (Lin-A), a clinically useful antibiotic to treat Gram-positive bacterial infections. Three methylation steps, catalyzed by three different S-adenosylmethionine (SAM)-dependent methyltransferases, are required in the biosynthesis of Lin-A, and thus highlight the significance of methyl group supply in lincomycin production. In this study, we demonstrate that externally supplemented SAM cannot be taken in by cells and therefore does not enhance Lin-A production. Furthermore, bioinformatics and in vitro enzymatic assays revealed there exist two SAM synthetase homologs, MetK1 (SLCG_1651) and MetK2 (SLCG_3830) in S. lincolnensis that could convert L-methionine into SAM in the presence of ATP. Even though we attempted to inactivate metK1 and metK2, only metK2 was deleted in S. lincolnensis LCGL, named as ΔmetK2. Following a reduction of the intracellular SAM concentration, ΔmetK2 mutant exhibited a significant decrease of Lin-A in comparison to its parental strain. Individual overexpression of metK1 or metK2 in S. lincolnensis LCGL either elevated the amount of intracellular SAM, concomitant with 15% and 22% increase in Lin-A production, respectively. qRT-PCR assays showed that overexpression of either metK1 or metK2 increased the transcription of lincomycin biosynthetic genes lmbA and lmbR, and regulatory gene lmbU, indicating SAM may also function as a transcriptional activator. When metK1 and metK2 were co-expressed, Lin-A production was increased by 27% in LCGL, while by 17% in a high-yield strain LA219X.


Assuntos
Antibacterianos/metabolismo , Lincomicina/metabolismo , Metionina Adenosiltransferase/metabolismo , Streptomyces/metabolismo , Proteínas de Bactérias/metabolismo , Regulação Bacteriana da Expressão Gênica , S-Adenosilmetionina , Metabolismo Secundário , Streptomyces/genética , Fatores de Transcrição
8.
J Ind Microbiol Biotechnol ; 45(6): 447-448, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29846840

RESUMO

In the online published article, row value "pIB139-metK1-metK2" in table 1 has been processed incorrectly. The correct table is given below.

9.
J Shoulder Elbow Surg ; 27(4): 599-605, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29241660

RESUMO

BACKGROUND: Previous studies have reported that glenohumeral internal rotation deficit (GIRD) may increase the risk of shoulder injury. However, the effects of GIRD on baseball pitching among pitchers of different age groups are still unclear. METHODS: The study participants were 24 high school and 24 university pitchers. For each age group, the pitchers were evenly divided into a GIRD group and a normal group. The pitching motion of each participant was captured using a motion analysis system at a sampling frequency of 300 Hz. The kinematics and kinetics of the throwing shoulder and trunk were quantified, and statistical differences between the groups were examined by 2-sample t tests. RESULTS: For both age groups, significant differences were observed in shoulder external rotations of the GIRD and normal groups. Compared with the university pitchers in the normal group, the university pitchers with GIRD exhibited a greater shoulder loading and did more internal rotation work in the acceleration phase. The high school pitchers with GIRD showed a larger trunk tilt and less trunk rotation than the university pitchers with GIRD. However, the university pitchers with GIRD exhibited a larger shoulder posterior force and horizontal adduction torque than the high school pitchers with GIRD. CONCLUSION: Pitchers with GIRD do change their pitching motions, and the greater resulting shoulder joint loading predisposes them to a greater risk of shoulder injury, especially among university pitchers.


Assuntos
Beisebol , Movimento , Rotação , Articulação do Ombro/fisiopatologia , Ombro/fisiopatologia , Tronco/fisiopatologia , Adolescente , Fatores Etários , Desempenho Atlético/fisiologia , Beisebol/lesões , Fenômenos Biomecânicos , Humanos , Masculino , Amplitude de Movimento Articular , Torque , Universidades , Adulto Jovem
10.
J Transl Med ; 15(1): 4, 2017 01 04.
Artigo em Inglês | MEDLINE | ID: mdl-28049487

RESUMO

BACKGROUND: We previously demonstrated that endothelial microparticles (EMPs) are increased in mitral valve diseases and impair valvular endothelial cell function. Perioperative systemic inflammation is an important risk factor and complication of cardiac surgery. In this study, we investigate whether EMPs increase in congenital heart diseases to promote inflammation and endothelial dysfunction. METHODS: The level of plasma EMPs in 20 patients with atrial septal defect (ASD), 23 patients with ventricular septal defect (VSD), and 30 healthy subjects were analyzed by flow cytometry. EMPs generated from human umbilical vascular endothelial cells (HUVECs) were injected into C57BL6 mice, or cultured with HUVECs without or with siRNAs targeting P38 MAPK. The expression and/or phosphorylation of endothelial nitric oxide synthase (eNOS), P38 MAPK, and caveolin-1 in mouse heart and/or in cultured HUVECs were determined. We evaluated generation of nitric oxide (NO) in mouse hearts, and levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in cultured HUVECs and in mice. RESULTS: EMPs were significantly elevated in patients with ASD and VSD, especially in those with pulmonary hypertension when compared with controls. EMPs increased caveolin-1 expression and P38 MAPK phosphorylation and decreased eNOS phosphorylation and NO production in mouse hearts. EMPs stimulated P38 MAPK expression, TNF-α and IL-6 production, which were all inhibited by siRNAs targeting P38 MAPK in cultured HUVECs. CONCLUSIONS: EMPs were increased in adult patients with congenital heart diseases and may contribute to increased inflammation leading to endothelial dysfunction via P38 MAPK-dependent pathways. This novel data provides a potential therapeutic target to address important complications of surgery of congenial heart disease.


Assuntos
Micropartículas Derivadas de Células/metabolismo , Células Endoteliais/metabolismo , Cardiopatias Congênitas/patologia , Cardiopatias Congênitas/fisiopatologia , Adulto , Animais , Caveolina 1/metabolismo , Demografia , Ecocardiografia Doppler , Endotélio Vascular/diagnóstico por imagem , Endotélio Vascular/patologia , Endotélio Vascular/fisiopatologia , Feminino , Cardiopatias Congênitas/sangue , Cardiopatias Congênitas/diagnóstico por imagem , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Interleucina-6/sangue , Masculino , Camundongos , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo III/metabolismo , Fosforilação , Fator de Necrose Tumoral alfa/sangue , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
11.
Metab Eng ; 39: 29-37, 2017 01.
Artigo em Inglês | MEDLINE | ID: mdl-27794466

RESUMO

Leucine-responsive regulatory proteins (Lrps) are a group of transcriptional regulators that regulate diverse cellular processes in bacteria and archaea. However, the regulatory role of Lrps in antibiotic biosynthesis remains poorly understood. In this study, we show that SACE_5388, an Lrp family regulator named as SACE_Lrp, is an efficient regulator for transporting and catabolizing branched-chain amino acids (BCAAs), playing an important role in regulating erythromycin production in Saccharopolyspora erythraea. SACE_Lrp directly controlled the expression of the divergently transcribed SACE_5387-5386 operon putatively encoding a BCAA ABC transporter by interacting with the intergenic region between SACE_Lrp and SACE_5387 (SACE_Lrp-5387-int), and indirectly controlled the expression of ilvE putatively encoding an aminotransferase catabolizing BCAAs. BCAA catabolism is one source of the precursors for erythromycin biosynthesis. Lysine and arginine promoted the dissociation of SACE_Lrp from SACE_Lrp -5387-int, whereas histidine increased their binding. Gene disruption of SACE_Lrp (ΔSACE_Lrp) in S. erythraea A226 resulted in a 25% increase in erythromycin production, while overexpression of SACE_5387-5386 in A226 enhanced erythromycin production by 36%. Deletion of SACE_Lrp (WBΔSACE_Lrp) in the industrial strain S. erythraea WB enhanced erythromycin production by 19%, and overexpression of SACE_5387-5386 in WBΔSACE_Lrp (WBΔSACE_Lrp/5387-5386) increased erythromycin production by 41% compared to WB. Additionally, supplement of 10mM valine to WBΔSACE_Lrp/5387-5386 culture further increased total erythromycin production up to 48%. In a 5-L fermenter, the erythromycin accumulation in the engineered strain WBΔSACE_Lrp/5387-5386 with 10mM extra valine in the industrial culture media reached 5001mg/L, a 41% increase over 3503mg/L of WB. These insights into the molecular regulation of antibiotic biosynthesis by SACE_Lrp in S. erythraea are instrumental in increasing industrial production of secondary metabolites.


Assuntos
Vias Biossintéticas/genética , Eritromicina/biossíntese , Melhoramento Genético/métodos , Proteína Reguladora de Resposta a Leucina/genética , Engenharia Metabólica/métodos , Saccharopolyspora/fisiologia , Proteínas de Bactérias/genética , Eritromicina/isolamento & purificação , Redes e Vias Metabólicas/genética
12.
Appl Microbiol Biotechnol ; 101(14): 5773-5783, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28601893

RESUMO

Lrp/AsnC family regulators have been found in many bacteria as crucial regulators controlling diverse cellular processes. By genomic alignment, we found that SCO3361, an Lrp/AsnC family protein from Streptomyces coelicolor, shared the highest similarity to the SACE_Lrp from Saccharopolyspora erythraea. Deletion of SCO3361 led to dramatic reduction in actinorhodin (Act) production and delay in aerial mycelium formation and sporulation on solid media. Dissection of the mechanism underlying the function of SCO3361 in Act production revealed that it altered the transcription of the cluster-situated regulator gene actII-ORF4 by directly binding to its promoter. SCO3361 was an auto-regulator and simultaneously activated the transcription of its adjacent divergently transcribed gene SCO3362. SCO3361 affected aerial hyphae formation and sporulation of S. coelicolor by activating the expression of amfC, whiB, and ssgB. Phenylalanine and cysteine were identified as the effector molecules of SCO3361, with phenylalanine reducing the binding affinity, whereas cysteine increasing it. Moreover, interactional regulation between SCO3361 and SACE_Lrp was discovered for binding to each other's target gene promoter in this work. Our findings indicate that SCO3361 functions as a pleiotropic regulator controlling secondary metabolism and morphological development in S. coelicolor.


Assuntos
Proteínas de Bactérias/metabolismo , Regulação Bacteriana da Expressão Gênica , Proteína Reguladora de Resposta a Leucina/genética , Streptomyces coelicolor/genética , Streptomyces coelicolor/metabolismo , Fatores de Transcrição/metabolismo , Antraquinonas/metabolismo , Proteínas de Bactérias/genética , Proteínas de Transporte , Cisteína/metabolismo , Deleção de Genes , Perfilação da Expressão Gênica , Proteína Reguladora de Resposta a Leucina/metabolismo , Família Multigênica , Fenilalanina/metabolismo , Regiões Promotoras Genéticas , Metabolismo Secundário , Fatores de Transcrição/genética
13.
Plant Foods Hum Nutr ; 72(1): 54-59, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-27924413

RESUMO

Fresh button mushrooms (Agaricus bisporus) were harvested and treated with a solution of 1.5% CaCl2 + 0.5% citric acid and stored for 16 days at 12 °C. The effects of this treatment on firmness, weight, color, cell wall compositions (cellulose and chitin) and cell wall degrading enzymes (cel1ulase, beta-1, 3 glucanase, chitinase and phenylalanine ammonialyase) were investigated during post-harvest storage. The expressions of major genes (Cel1, Glu1, Chi1 and PAL1) involved in cell wall degradation during post-harvest storage were also monitored. The results revealed that the post-harvest chemical treatment maintained better firmness, weight, color and inhibited cellulase, beta-1, 3 glucanase, chitinase and phenylalanine ammonialyase activities. These findings showed that the down-regulation of cell wall degrading enzymes is a possible mechanism that delays the softening of button mushrooms by the application of combined chemical treatment.


Assuntos
Agaricus/efeitos dos fármacos , Cloreto de Cálcio/farmacologia , Ácido Cítrico/farmacologia , Conservação de Alimentos/métodos , Agaricus/enzimologia , Agaricus/genética , Parede Celular/efeitos dos fármacos , Parede Celular/enzimologia , Regulação para Baixo , Fatores de Tempo
14.
15.
Appl Microbiol Biotechnol ; 100(5): 2257-66, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26552796

RESUMO

Glycosyltransferase DesVII and its auxiliary partner DesVIII from Streptomyces venezulae, homologs of EryCIII and EryCII in Saccharopolyspora erythraea, have previously been demonstrated to be flexible on their substrates in vitro. Herein, we investigated their in vivo function by interspecies complementation in the mutant strains of Sac. erythraea A226. As desVII and desVIII were concomitantly expressed in the ΔeryCIII mutant, the erythromycin A (Er-A) production was restored. Interestingly, co-expression of desVII and desVIII in the ΔeryBV mutant exhibited an increased Er-A yield by 15 % in comparison to A226. Hence, DesVII/DesVIII not only replaced EryCIII to upload D-desosamine to C5 position of 3-O-mycarosyl erythronolide B (MEB) but also in vivo attached L-mycarose, not D-desosamine to C3 position of erythronolide B (EB) with a higher activity than EryBV. Furthermore, expression of desVII in ΔeryCIII and ΔeryBV-CIII partially restored the Er-A production; however, no Er-A was detected while desVII was expressed in ΔeryBV. It was implicated that DesVII coupled with EryCII to form the DesVII/EryCII complex for attaching above two deoxysugars in the absence of EryCIII in Sac. erythraea. In addition, when desVII and desVIII were co-expressed in ΔeryBV-CII, Er-A was recovered with a lower yield than ΔeryBV-CIII. Our study presents an opportunity with Sac. erythraea as a cell factory for macrolide glycodiversification.


Assuntos
Antibacterianos/metabolismo , Eritromicina/metabolismo , Glicosiltransferases/metabolismo , Saccharopolyspora/enzimologia , Saccharopolyspora/metabolismo , Streptomyces/enzimologia , Streptomyces/metabolismo , Técnicas de Inativação de Genes , Teste de Complementação Genética , Glicosiltransferases/genética , Streptomyces/genética
16.
Med Sci Monit ; 22: 460-8, 2016 Feb 12.
Artigo em Inglês | MEDLINE | ID: mdl-26868022

RESUMO

BACKGROUND Colon adenocarcinoma mostly happens at the junction of the rectum and is a common gastrointestinal malignancy. Accumulated evidence has indicated that colon adenocarcinoma develops by genetic alterations and is a complicated disease. The aim of this study was to screen differentially expressed miRNAs (DEMs) and genes with diagnostic and prognostic potentials in colon adenocarcinoma. MATERIAL AND METHODS In this study we screened DEMs and their target genes (DEGs) between 100 colon adenocarcinoma and normal samples in The Cancer Genome Atlas (TCGA) database by using the DEseq toolkit in Bioconductor. Then Go enrichment and KEGG pathway analysis were performed on the selected differential genes by use of the DAVID online tool. A regulation network of miRNA-gene was constructed and analyzed by Cytoscape. Finally, we performed ROC analysis of 8 miRNAs and ROC curves were drawn. RESULTS A total of 159 DEMs and 1921 DEGs were screened, and 1881 pairs of miRNA-target genes with significant negative correlations were also obtained. A regulatory network of miRNA-gene, including 60 cancer-related genes and 47 miRNAs, was successfully constructed. In addition, 5 clusters with several miRNAs regulating a set of target genes simultaneously were identified through cluster analysis. There were 8 miRNAs involved in these 5 clusters, and these miRNAs could serve as molecular biomarkers to distinguish colon adenocarcinoma and normal samples indicated by ROC analysis. CONCLUSIONS The identified 8 miRNAs were closely associated with colon adenocarcinoma, which may have great clinical value as diagnostic and prognostic biomarkers and provide new ideas for targeted therapy.


Assuntos
Adenocarcinoma/genética , Neoplasias do Colo/genética , MicroRNAs/genética , Adenocarcinoma/diagnóstico , Adenocarcinoma/metabolismo , Biomarcadores Tumorais/biossíntese , Biomarcadores Tumorais/genética , Análise por Conglomerados , Neoplasias do Colo/diagnóstico , Neoplasias do Colo/metabolismo , Perfilação da Expressão Gênica , Humanos , MicroRNAs/biossíntese , Prognóstico , Curva ROC
17.
Mol Med ; 21: 134-42, 2015 Jan 12.
Artigo em Inglês | MEDLINE | ID: mdl-25587856

RESUMO

The host tolerance mechanisms to avian influenza virus (H5N1) infection that limit tissue injury remain unknown. Emerging evidence indicates that cystic fibrosis transmembrane conductance regulator (CFTR), a cAMP-dependent Cl(-) channel, modulates airway inflammation. Janus tyrosine kinase (JAK) 3, a JAK family member that plays a central role in inflammatory responses, prominently contributes to the dysregulated innate immune response upon H5N1 attachment; therefore, this study aims to elucidate whether JAK3 activation induced by H5N1 hemagglutinin (HA) inhibits cAMP-dependent CFTR channels. We performed short-circuit current, immunohistochemistry and molecular analyses of the airway epithelium in Jak3(+/+) and Jak3(+/-) mice. We demonstrate that H5N1 HA attachment inhibits cAMP-dependent CFTR Cl(-) channels via JAK3-mediated adenylyl cyclase (AC) suppression, which reduces cAMP production. This inhibition leads to increased nuclear factor-kappa B (NF-κB) signaling and inflammatory responses. H5N1 HA is detected by TLR4 expressed on respiratory epithelial cells, facilitating JAK3 activation. This activation induces the interaction between TLR4 and Gαi protein, which blocks ACs. Our findings provide novel insight into the pathogenesis of acute lung injury via the inhibition of cAMP-dependent CFTR channels, indicating that the administration of cAMP-elevating agents and targeting JAK3 may activate host tolerance to infection for the management of influenza virus-induced fatal pneumonia.


Assuntos
AMP Cíclico/metabolismo , Regulador de Condutância Transmembrana em Fibrose Cística/metabolismo , Glicoproteínas de Hemaglutininação de Vírus da Influenza/metabolismo , Virus da Influenza A Subtipo H5N1/fisiologia , Janus Quinase 3/metabolismo , Pneumonia Viral/metabolismo , Pneumonia Viral/virologia , Receptor 4 Toll-Like/metabolismo , Animais , Regulador de Condutância Transmembrana em Fibrose Cística/genética , Modelos Animais de Doenças , Ativação Enzimática , Expressão Gênica , Humanos , Influenza Humana/metabolismo , Influenza Humana/virologia , Janus Quinase 3/genética , Camundongos , Camundongos Knockout , NF-kappa B/metabolismo , Pneumonia Viral/patologia , Ligação Proteica , Mucosa Respiratória/metabolismo , Mucosa Respiratória/patologia , Mucosa Respiratória/virologia , Transdução de Sinais
18.
Appl Microbiol Biotechnol ; 98(21): 9009-20, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25236801

RESUMO

Lysobacter enzymogenes is a bacterial biological control agent emerging as a new source of antibiotic metabolites, such as heat-stable antifungal factor (HSAF) and the antibacterial factor WAP-8294A2. The regulatory mechanism(s) for antibiotic metabolite biosynthesis remains largely unknown in L. enzymogenes. Clp, a cyclic adenosine monophosphate (cAMP)-receptor-like protein, is shown to function as a global regulator in modulating biocontrol-associated traits in L. enzymogenes. However, the genetic basis of Clp signaling remains unclear. Here, we utilized transcriptome/microarray analysis to determine the Clp regulon in L. enzymogenes. We showed that Clp is a global regulator in gene expression, as the transcription of 775 genes belonging to 19 functional groups was differentially controlled by Clp signaling. Analysis of the Clp regulon detected previously characterized Clp-modulated functions as well as novel loci. These include novel loci involved in antibiotic metabolite biosynthesis and surface motility in L. enzymogenes. We further showed experimentally that Clp signaling played a positive role in regulating the biosynthesis of HSAF and WAP-8294A2, as well as surface motility which is a type-IV-pilus-dependent trait. The regulation by Clp signaling of antibiotic (HSAF and WAP-8294A2) biosynthesis and surface motility was found to be independent. Importantly, we identified a factor Lysobacter acetyltransferase (Lat), a homologue of histone acetyltransferase Hpa2, which was regulated by Clp and involved in HSAF biosynthesis, but not associated with WAP-8294A2 production and surface motility. Overall, our study provided new insights into the regulatory role and molecular mechanism of Clp signaling in L. enzymogenes.


Assuntos
Anti-Infecciosos/metabolismo , Regulação Bacteriana da Expressão Gênica , Locomoção , Lysobacter/fisiologia , Metabolismo Secundário , Transdução de Sinais , Fatores de Transcrição/metabolismo , Perfilação da Expressão Gênica , Lysobacter/genética , Análise em Microsséries
19.
J Sci Food Agric ; 94(15): 3273-80, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-24700113

RESUMO

BACKGROUND: In order to evaluate the effects of nano-CaCO3 -based low density polyethylene (nano-CaCO3 -LDPE) packaging on the quality of fresh-cut sugarcane, concentrations of O2 and CO2 within the packages, overall visual quality (OVQ), total bacterial count (TBC), yeast and mould count (YMC), reducing sugar content and total phenolic content, respiration, ethylene production, and the activities of phenylalanine ammonia-lyase (PAL), polyphenol oxidase (PPO), peroxidase (POD), acid invertase (AI) and neutral invertase (NI) were examined during storage at 10 °C for 5 days. RESULTS: The transmission rate of O2 and CO2 of the nano-CaCO3 -LDPE material was lower than that of LDPE, which lead to the more rapid formation of gas environment with low O2 and high CO2 concentration in the package. TBC and YMC counts of fresh-cut sugarcane were significantly retarded by nano-CaCO3 -LDPE packaging. Nano-CaCO3 -LDPE packaging fresh-cut sugarcane exhibited significantly lower activities of PAL, PPO, POD AI and NI than LDPE packaging fresh-cut sugarcanes during the storage. Meanwhile, nano-CaCO3 -LDPE packaging significantly inhibited the increase of browning index and total phenolic content, while improving OVQ. CONCLUSION: Our results indicated that nano-CaCO3 -LDPE packaging together with the cold storage is a promising approach in inhibiting browning and maintaining quality of fresh-cut sugarcane.


Assuntos
Carbonato de Cálcio , Embalagem de Alimentos/instrumentação , Qualidade dos Alimentos , Nanoestruturas , Polietileno , Saccharum , Carga Bacteriana , Dióxido de Carbono/análise , Catecol Oxidase/metabolismo , Temperatura Baixa , Contagem de Colônia Microbiana , Embalagem de Alimentos/métodos , Conservação de Alimentos/métodos , Reação de Maillard , Oxigênio/análise , Peroxidase/metabolismo , Fenóis/análise , Fenilalanina Amônia-Liase/metabolismo , Saccharum/microbiologia , beta-Frutofuranosidase/metabolismo
20.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 45(3): 529-32, 2014 May.
Artigo em Zh | MEDLINE | ID: mdl-24941833

RESUMO

OBJECTIVE: To analyze the efficacy and safety of subcutaneous administration of bortezomib in the treatment of multiple myeloma (MM) patients. METHODS: A total of 26 MM patients were enrolled in this study and treated with BDT (bortezomib-dexamethasone-thalidomide). In the 26 MM patients, 12 patients received subcutaneous administration of Bortezomib while 14 patients received conventional intravenous administration. The outcomes and adverse effects of two groups were retrospectively evaluated and compared. RESULTS: Overall response (OR) rates in the two groups were 75.00% and 71.43% respectively, in which complete remission (CR) plus very good complete remission (VGPR) rates were 50.00% and 47.14%, while CR rates were 16.67% and 28.57%. There were no statistically significant difference (P > 0.05). Time to achieve effectiveness in two groups was similar (P > 0.05). More than half patients in both groups achieved partial remission after the first treatment course and CR after the fourth course. Compared to the intravenous group, peripheral neuropathy rates remained significantly lower in subcutaneous group (16.67% vs. 64.29%, P = 0.021). The intravenous group had 7.14% grade 3 or worse, peripheral neuropathy but none found in the subcutaneous group. Rash occurred only in subcutaneous group (66.67%), but it was local, mild and transitional. No significant differences of other adverse events between the two groups were observed. CONCLUSION: Subcutaneous administration of bortezomib offers similar efficacy to standard intravenous administration in the treatment of multiple myelom, with an improved safety for lower rate of peripheral neuropathy.


Assuntos
Ácidos Borônicos/administração & dosagem , Mieloma Múltiplo/tratamento farmacológico , Pirazinas/administração & dosagem , Indução de Remissão , Protocolos de Quimioterapia Combinada Antineoplásica , Ácidos Borônicos/uso terapêutico , Bortezomib , Dexametasona , Humanos , Infusões Intravenosas , Injeções Subcutâneas , Pirazinas/uso terapêutico , Estudos Retrospectivos
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