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1.
J Physiol Sci ; 74(1): 2, 2024 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-38166513

RESUMO

Establishing specific reference intervals (RIs) of serum 25-hydroxyvitamin D3 [25(OH)D] for children is essential for improving the accuracy of diagnosis and prognosis monitoring of diseases such as rickets and growth retardation. The study including 6,627 healthy children was conducted to establish specific RIs of 25(OH)D for children in Nanning area of China. The results showed that there were statistically significant differences among age, season, and gender of serum 25(OH)D levels, and the age-specific RIs of serum 25(OH)D were 20.3 ~ 53.6 ng/mL for 0 ~ ≤ 1 year and 18.9 ~ 49.6 ng/mL for 2 ~ ≤ 3 years. The age-, season-specific RIs of serum 25(OH)D for 4 ~ ≤ 6 years in spring-summer and autumn-winter were 15.8 ~ 42.6 ng/mL and 15.2 ~ 37.7 ng/mL, respectively. The age-, gender-specific RIs of serum 25(OH)D for 7 ~ ≤ 18 years for males and females were 12.1 ~ 36.1 ng/mL and 10.8 ~ 35.3 ng/mL, respectively. This study successfully established the RIs of serum 25(OH)D, which may help to improve disease diagnosis and monitoring for children in the Nanning area of China.


Assuntos
Calcifediol , Vitamina D , Masculino , Criança , Feminino , Humanos , Adolescente , Estações do Ano , China
2.
J Microbiol Immunol Infect ; 55(6 Pt 2): 1246-1254, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34924339

RESUMO

BACKGROUND/PURPOSE: Biofilms formed by Klebsiella pneumoniae on medical devices increase infection risk. Fimbriae and capsule polysaccharides (CPSs) are important factors involved in biofilm formation. KP1_4563 in K. pneumoniae NTUH-K2044, a small protein containing the DUF1471 domain, was reported to inhibit type 3 fimbriae function. In this study, we aimed to determine whether the KP1_4563 homolog is conserved in each K. pneumoniae isolate and what role it has in Klebsiella biofilms. METHODS: The genomes of K. pneumoniae NTUH-K2044, CG43, MGH78578, KPPR1 and STU1 were compared. The KP1_4563 homolog in K. pneumoniae STU1 was named orfX. Biofilms of wild-type and orfX mutant strains from K. pneumoniae STU1 and one clinical isolate, 83535, were quantified. Transcription levels of the type 3 fimbrial genes, mrkA and mrkH, were investigated by RT-qPCR. MrkA of the wild-type and orfX mutant were observed by Western blotting. The morphology of bacterial cells was observed by transmission electron microscopy (TEM). Bacterial CPSs were quantified. RESULTS: The gene and upstream region of orfX were conserved among the five K. pneumoniae isolates. Deletion of orfX enhanced Klebsiella biofilm formation. However, the amount of mRNA from mrkA and mrkH and the level of MrkA protein were not different between the wild type and orfX mutant. In contrast, the amount of CPS in orfX mutants was increased, compared to their parental strains, STU1 and 83535. CONCLUSION: The role of orfX is speculated to be conserved in most K. pneumoniae isolates. OrfX negatively controlled biofilm formation by reducing CPS, not type 3 fimbriae, production.


Assuntos
Infecções por Klebsiella , Klebsiella pneumoniae , Humanos , Klebsiella pneumoniae/genética , Klebsiella pneumoniae/metabolismo , Regulação Bacteriana da Expressão Gênica , Biofilmes , Fímbrias Bacterianas/genética , Fímbrias Bacterianas/metabolismo , Infecções por Klebsiella/microbiologia
3.
Bioresour Technol ; 302: 122889, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32033841

RESUMO

The increasing amounts of food wastage and accumulation generated per annum due to the growing human population worldwide often associated with environmental pollution issues and scarcity of natural resources. In view of this, science community has worked towards in finding sustainable approaches to replace the common practices for food waste management. The agricultural and food processing wastes rich in nutrients are often the attractive substrates for the bioconversion for valuable bioproducts such as industrial enzymes, biofuel and bioactive compounds. The sustainable approaches on the re-utilization of food wastes as the industrial substrates for production of valuable bioproducts has meet the goals of circular bioeconomy, results in the diversify applications and increasing market demands for the bioproducts. This review discusses the current practice and recent advances on reutilization of food waste for bioconversion of valuable bioproducts from agricultural and food processing wastes.


Assuntos
Eliminação de Resíduos , Gerenciamento de Resíduos , Agricultura , Biocombustíveis , Alimentos
4.
Appl Biochem Biotechnol ; 191(1): 346-359, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-31863348

RESUMO

1,3-Propanediol (1,3-PDO) has numerous industrial applications in the synthesis of the monomer of the widely used fiber polytrimethylene terephthalate. In this work, the production of 1,3-PDO by Klebsiella pneumoniae is increased by dual-substrate cultivation and fed-batch fermentation. Experimental results indicate that the production of 1,3-PDO can be elevated to 16.09 g/L using a dual substrate ratio (of glucose to crude glycerol) of 1/30 and to 20.73 g/L using an optimized dual-substrate ratio of 1/20. Ultimately, the optimal dual-substrate feeding for a 5 L scale fed-batch fermenter that maximizes 1,3-PDO production (29.69 g/L) is determined. This production yield is better than that reported in most related studies. Eventually, the molecular weight and chemical structure of 1,3-PDO were obtained by FAB-MS, 1H-NMR, and 13C-NMR. Also, in demonstrating the effectiveness of the fermentation strategy in increasing the production and production yield of 1,3-PDO, experimental results indicate that the fermentation of 1,3-PDO is highly promising for commercialization.


Assuntos
Klebsiella pneumoniae/crescimento & desenvolvimento , Propilenoglicóis/metabolismo , Glucose/metabolismo , Glicerol/metabolismo
5.
Prep Biochem Biotechnol ; 50(1): 74-81, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31517565

RESUMO

Ectoine has fostered the development of products for skin care and cosmetics. In this study, we employed the marine bacterial strain Marinococcus sp. MAR2 to increase ectoine production by optimizing medium constituents using Response Surface Methodology (RSM) and a fed-batch strategy. The results from the steepest ascent and central composite design indicated that 54 g/L of yeast extract, 14.0 g/L of ammonium acetate, 74.4 g/L of sodium glutamate, and 6.2 g/L of sodium citrate constituted the optimal medium with maximum ectoine production (3.5 g/L). In addition, we performed fed-batch culture in the bioreactor, combining pH and dissolved oxygen to produce ectoine by Marinococcus sp. MAR2. The ectoine production, content, and productivity of 5.6 g/L, 10%, and 3.9 g/L/day were further reached by a fed-batch culture. Thus, the ectoine production by Marinococcus sp. MAR2 using RSM and fed-batch strategy shows its potential for industrial production.


Assuntos
Diamino Aminoácidos/metabolismo , Bacillaceae/metabolismo , Técnicas de Cultura Celular por Lotes/métodos , Microbiologia Industrial/métodos , Acetatos/análise , Acetatos/metabolismo , Bacillaceae/crescimento & desenvolvimento , Técnicas de Cultura Celular por Lotes/instrumentação , Reatores Biológicos , Meios de Cultura/química , Meios de Cultura/metabolismo , Desenho de Equipamento , Fermentação , Microbiologia Industrial/instrumentação , Citrato de Sódio/análise , Citrato de Sódio/metabolismo , Glutamato de Sódio/análise , Glutamato de Sódio/metabolismo
6.
Bioresour Technol ; 291: 121891, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31387049

RESUMO

This work studies a series of strategies in the production of lutein by Scenedesmus obliquus CWL-1 under mixotrophic cultivation. Our experimental results revealed that the optimal conditions associated with light-related strategies were 12 h light period followed by a 12 h dark period and blue to red light under mixotrophic cultivation. Under such conditions, the biomass, lutein content and lutein productivity were maximized to 9.88 (g/L), 1.78 (mg/g) and 1.43 (mg/L/day), respectively. Moreover, the assimilation of 4.5 g/L of calcium nitrate into S. obliquus CWL-1 increased the maximal biomass (12.73 g/L) and the highest maximal lutein productivity (3.06 mg/L/day), while the assimilation of 1.5 g/L of calcium nitrate yielded the highest maximal lutein content of 2.45 mg/g. The highest maximal lutein productivity of 4.96 (mg/L/day) was obtained when fed-batch fermentation was conducted, and this value was approximately 11-folds that obtained using the batch system.


Assuntos
Luteína/biossíntese , Microalgas/metabolismo , Scenedesmus/metabolismo , Biomassa , Fermentação , Luz
7.
J Biosci Bioeng ; 128(3): 332-336, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30935782

RESUMO

Hydroxyectoine, an ectoine derivative, is the most common compatible solute in halophilic microorganisms for resisting harsh environments. Compatible solutes can be utilized in fields such as cosmetics, medicine, and biochemistry. Moderately halophilic microorganisms produce much less hydroxyectoine as compared with ectoine. In this study, we first evaluate the effect of medium formulation (i.e., yeast extract (YE) medium and high yeast extract (HYE) medium) on hydroxyectoine production. In addition, an investigation of hydroxyectoine production by Halomonas salina under optimal conditions for vital factors (i.e., iron and α-ketoglutarate) and hydroxylase activity was also carried out. As a result, hydroxyectoine production was obviously elevated (0.9 g/L to 1.8 g/L) when the HYE medium was utilized. Furthermore, hydroxyectoine production further increased to 2.4 g/L when both the α-ketoglutarate and iron factors were added to the HYE medium in the early stationary phase. In addition, we found that ectoine hydroxylase activity increased more when a combination of iron and α-ketoglutarate was used than when either was used alone. The results showed that the alteration of iron and α-ketoglutarate clearly stimulated the expression of ectoine hydroxylase, which in turn affected hydroxyectoine synthesis. This study also showed that hydroxyectoine production was further raised from 2.4 g/L to 2.9 g/L when 50 mM of α-ketoglutarate and 1 mM of iron were added to the HYE medium. Ultimately, the experimental results showed using the optimal conditions further elevated the hydroxyectoine production yield to 2.90 g/L, which was over 3-fold higher than the best results obtained from the original medium.


Assuntos
Diamino Aminoácidos/metabolismo , Fermentação/fisiologia , Halomonas/metabolismo , Reatores Biológicos/microbiologia , Meios de Cultura/química , Halomonas/enzimologia , Ferro/química , Ácidos Cetoglutáricos/química , Engenharia Metabólica/métodos , Técnicas Microbiológicas/métodos , Oxigenases de Função Mista/genética , Oxigenases de Função Mista/metabolismo , Tolerância ao Sal
8.
Sci Total Environ ; 657: 871-881, 2019 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-30677952

RESUMO

Soil dissolved organic matter (DOM) alters heavy metal availability, but whether straw amendment can manipulate soil selenium (Se) speciation and availability through DOM mineralization remains unclear. In this study, allochthonous maize straw and selenate were incubated together in four different soils for 1 y. The transformation and availability of DOM associated Se (DOM-Se) was investigated during aging. Results indicated that soil solution and soil particle surfaces were dominated by hexavalent hydrophilic acid-bound Se (Hy-Se). The amount of fulvic acid bound Se in soil solution (SOL-FA-Se) was higher than humic acid bound Se in soil solution (SOL-HA-Se), except in krasnozems, and mainly existed as hexavalent Se (Se(VI)). Tetravalent Se (Se(IV)) was the main valence state of FA-Se adsorbed on soil particle surfaces (EX-FA-Se) after 5 w of aging. The proportion of soil-available Se (SOL + EX-Se) decreased with increasing straw rate. However, under an application rate of 7500 kg·hm-2, soluble Se fraction (SOL-Se) reduction was minimal in acidic soils (18.7%-34.7%), and the organic bound Se fraction (OM-Se) was maximally promoted in alkaline soils (18.2%-39.1%). FA and HON could enhance the availability of Se in the soil solution and on particle surfaces of acidic soil with high organic matter content. While Se incorporation with HA could accelerate the fixation of Se into the solid phase of soil. Three mechanisms were involved in DOM-Se aging: (1) Reduction, ligand adsorption, and inner/outer-sphere complexation associated with the functional groups of straw-derived DOM, including hydroxyls, carboxyl, methyl, and aromatic phenolic compounds; (2) interconnection of EX-FA-Se between non-residual and residual Se pools; and (3) promotion by soil electrical conductivity (EC), clay, OM, and straw application. The dual effect of DOM on Se aging was highly reliant on the characteristics of the materials and soil properties. In conclusion, straw amendment could return selenium in soil and reduce soluble Se loss.


Assuntos
Recuperação e Remediação Ambiental/métodos , Selênio/análise , Solo/química , Agricultura , Benzopiranos/química , China , Interações Hidrofóbicas e Hidrofílicas , Caules de Planta/química , Ácido Selênico/química , Selênio/química , Selênio/farmacocinética , Poluentes do Solo/análise , Poluentes do Solo/farmacocinética , Espectroscopia de Infravermelho com Transformada de Fourier , Fatores de Tempo
9.
Nanoscale Adv ; 1(12): 4745-4754, 2019 Dec 03.
Artigo em Inglês | MEDLINE | ID: mdl-36133126

RESUMO

In recent years, flexible light-emitting devices (LEDs) have become the main focus in the field of display technology. Graphene, a two-dimensional layered material, has attracted great interest in LEDs due to its excellent properties. However, there are many problems such as efficiency, lifetime, and flexibility not well solved. Herein, we have successfully prepared a flexible LED using laser-induced reduced graphene oxide (LIRGO). The LIRGO LED achieves a luminescence lifetime of over 60 hours and a wall plug efficiency of up to 1.4% in a vacuum environment of 0.02 Pa. There are many small luminescent spots randomly distributed on 3.5 × 5 mm2 of LIRGO. LIRGO's luminous behavior can be controlled by modifying the supply voltage and laser reduction intensity. We also explore LIRGO's applications by testing it in different packages and customizable bulbs. Furthermore, as an interesting demo, the LIRGO device can be used to mimic constellations with visual shapes. This work demonstrates LIRGO's great potential in many fields, such as flexible and miniature light sources and displays.

10.
J Biosci Bioeng ; 126(6): 783-789, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30401454

RESUMO

The biosynthesis of prodigiosin (PG) from Serratia marcescens involves the coupling of a bipyrrole, 4-methoxy-2,2'-bipyrrole-5-carboxaldehyde (MBC), with a monopyrrole, 2-methyl-3-n-amyl-pyrrole (MAP), and formation of a linear tripyrrole (PG). We constructed mutant strains in which either the MBC biosynthesis by S. marcescens BMJ816 or the MAP biosynthesis by S. marcescens AMJ817. S. marcescens BMJ816 and AMJ817 confirmed that they lose the ability to synthesize PG when they are cultivated alone. An experiment was also conducted in which cultures of the two mutant strains were grown to the early exponential phase in a semi-defined medium, and one suspension culture was inoculated with the other. This approach yielded 103 mg/L PG. The findings suggest that the addition of precursors may enhance PG production by microorganisms.


Assuntos
Técnicas Microbiológicas/métodos , Prodigiosina/biossíntese , Serratia marcescens/genética , Serratia marcescens/metabolismo , Técnicas de Cocultura , Técnicas de Transferência de Genes , Organismos Geneticamente Modificados , Pirróis/metabolismo , Transformação Bacteriana
11.
J Biosci Bioeng ; 125(5): 578-584, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29331525

RESUMO

This study attempted to utilize Halomonas salina BCRC17875 to produce ectoine by optimizing the agitation speed and medium composition. In addition, the chemical structure of ectoine produced by H. salina BCRC17875 was determined. The results indicate that ectoine production reached 3.65 g/L at 38 h of cultivation when the agitation rate and NaCl concentration were fixed at 200 rpm and 2.0 M, respectively. It reached 9.20 g/L at 44 h of cultivation when the major medium components were yeast extract (56 g/L), glutamate (74.40 g/L), and ammonium sulfate (14 g/L). After the nitrogen concentration had been evaluated, evaluation of the nitrogen concentration revealed that the ectoine production reached 11.80 g/L at 44 h of cultivation when 56 g/L of yeast extract and 28 g/L of ammonium sulfate were used. Ectoine production reached 13.96 g/L at 44 h of cultivation when the carbon/nitrogen ratio was fixed at 3/1 using 84 g/L of yeast extract and 28 g/L of ammonium sulfate. Furthermore, the identification of ectoine were identified and characterized by fast atom bombardment mass spectrometry (FAB-MS) and 1H NMR. The results demonstrated a fermentation strategy was successful in increasing ectoine production, and that the fermentation medium of ectoine had commercialization potential.


Assuntos
Diamino Aminoácidos/metabolismo , Técnicas de Cultura Celular por Lotes/métodos , Halomonas/citologia , Halomonas/metabolismo , Diamino Aminoácidos/química , Técnicas Bacteriológicas/métodos , Carbono/metabolismo , Fermentação , Nitrogênio/metabolismo , Cloreto de Sódio/química
12.
Biomed Res Int ; 2018: 2710892, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30662902

RESUMO

Thermosensitive hydrogels are attractive alternative scaffolding materials for minimally invasive surgery through a simple injection and in situ gelling. In this study, a novel poly(ester-amide) polymer, methoxy poly(ethylene glycol)-poly(pyrrolidone-co-lactide) (mPDLA, P3L7) diblock copolymer, was synthesized and characterized for cartilage tissue engineering. A series of amphiphilic diblock copolymers was synthesized by ring-opening polymerization of mPEG 550, D,L-lactide, and 2-pyrrolidone. By dynamic light scattering analysis and tube-flipped-upside-down method, viscoelastic properties of the mPDLA diblock copolymer solution exhibited sol-gel transition behavior as a function of temperature. An in vitro degradation assay showed that degradation acidity was effectively reduced by introducing the 2-pyrrolidone monomer into the polyester hydrogel. Besides, mPDLA exhibited great biocompatibility in vitro for cell encapsulation due to a high swelling ratio. Moreover, cell viability and biochemical analysis proved that the mPDLA hydrogel presented a great chondrogenic response. Taken together, these results demonstrate that mPDLA hydrogels are promising injectable scaffolds potentially applicable to cartilage tissue engineering.


Assuntos
Plásticos Biodegradáveis/química , Cartilagem/química , Hidrogéis/química , Poliaminas/química , Poliésteres/química , Polímeros/química , Animais , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Dioxanos/química , Polietilenoglicóis/química , Coelhos , Temperatura , Engenharia Tecidual/métodos
13.
Bioresour Technol ; 245(Pt B): 1455-1460, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28596072

RESUMO

The aim of this work was to study the feasibility of surface displaying synthetic phytochelatin (EC) on Saccharomyces cerevisiae to overcome the inhibitory effect of heavy metals on ethanol production. Via the fusion of a gene encoding EC to an α-agglutinin gene, the engineered S. cerevisiae was able to successfully display EC on its surface. This surface engineered yeast strain exhibited an efficient cadmium adsorption capability and a remarkably enhanced cadmium tolerance. Moreover, its ethanol production efficiency was significantly improved as compared to a control strain in the presence of cadmium. Similar results could also be observed in the presence of other metals, such as nickel, lead and copper. Overall, this method allows simultaneous biorefinery and heavy metal removal when using heavy metal-contaminated biomass as raw materials.


Assuntos
Metais Pesados , Fitoquelatinas , Saccharomyces cerevisiae , Biodegradação Ambiental , Biomassa , Cádmio , Etanol
14.
J Biosci Bioeng ; 124(2): 184-188, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28325660

RESUMO

The aim of this study was to use a modified bioreactor system for simultaneous saccharification of cellulose and bioethanol production. We tested Aspergillus niger and Trichoderma reesei for cellulose saccharification and Zymomonas mobilis for bioethanol production simultaneously in this modified bioreactor. The results showed that various carboxymethylcellulose (CMC) concentrations (10, 15, or 20 g/L) as a substrate for A. niger and T. reesei yielded bioethanol production of 0.51, 0.78, and 0.89 g/L and a CMC conversion rate of 10.2%, 10.7%, and 8.89%, respectively. These data suggested that at 10 g/L CMC as a substrate, the CMC conversion rate was higher than that at the other concentrations. When CMC concentration exceeded 15 g/L, bioethanol production was prolonged to 40 h. These results were attributed to the viscosity of CMC. This study also tested Napier grass (an agricultural byproduct) for bioethanol production. The results revealed bioethanol production and the theoretical Napier grass conversion rate were 0.38 g/L and 12.6%, respectively, for 13-h cultivation when the feeding concentration of Napier grass was 10 g/L. When Napier grass concentration was increased to 15 g/L, bioethanol production and the Napier grass conversion rate reached 0.51 g/L and 23%, respectively, after 14-h cultivation. Eventually, the experimental results indicated using agricultural waste for bioethanol production has been become a potential strategy.


Assuntos
Aspergillus niger/metabolismo , Etanol/metabolismo , Pennisetum/microbiologia , Trichoderma/metabolismo , Zymomonas/metabolismo , Reatores Biológicos/microbiologia , Celulose/metabolismo , Etanol/análise , Fermentação , Hidrólise , Pennisetum/química , Pennisetum/metabolismo
15.
Artif Organs ; 38(6): 484-92, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24571555

RESUMO

Antibiotic-loaded acrylic bone cement has been frequently used as an infection prophylaxis or antibiotic-loaded spacer in infected arthroplasty. In addition, daptomycin has been used recently against broad spectrum Gram-positive organisms. The goal of this in vitro study is to investigate the bacteriacidal and mechanical properties of daptomycin-incorporated polymethylmethacrylate (PMMA) bone cement and evaluate its feasibility for clinical use. Daptomycin (0.5, 1, or 2 g) was premixed with 40 g of PMMA bone cement powder before curing. The mechanical properties of the daptomycin-loaded acrylic bone cement (DLABC) were estimated following standard guidance, and the release profile and kinetics of daptomycin from PMMA were analyzed. The antimicrobial efficacy of DLABC was determined with a zone of inhibition (ZOI) assay against Staphylococcus aureus, Staphylococcus epidermis, Enterococcus faecalis, and Enterococcus faecium, respectively. The results showed that the compressive strength, of PMMA bone cement, which was higher than 100 MPa in all groups, was sufficient according to ISO 5833 after incorporation of daptomycin. The encapsulated daptomycin was released for 2 weeks with a 9.59 ± 0.85%, 15.25 ± 0.69%, and 20.64 ± 20.33% released percentage on the first day in the low, mid, and high groups, respectively. According to the calculated release kinetics, incorporated daptomycin should be 3.3 times the original dose to double its release. Although all recipes of DLABC had a microbial inhibitory effect, the effect with a higher encapsulated amount of daptomycin was more significant. Therefore, we believe that daptomycin can be locally delivered from PMMA bone cement at the surgical site as a prophylactic or treatment for osteomyelitis against Gram-positive organisms with intact cement function.


Assuntos
Antibacterianos/farmacologia , Artroplastia/métodos , Bactérias/efeitos dos fármacos , Cimentos Ósseos/química , Daptomicina/farmacologia , Portadores de Fármacos , Polimetil Metacrilato/química , Antibacterianos/química , Artroplastia/efeitos adversos , Bactérias/crescimento & desenvolvimento , Força Compressiva , Daptomicina/química , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Estudos de Viabilidade , Cinética , Teste de Materiais , Solubilidade
16.
J Tissue Eng Regen Med ; 7(8): 665-72, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22408003

RESUMO

This study evaluates the crosslinkage effect of chondroitin sulphate C (CSC) and type II collagen (COL II) on chondrogenesis of mesenchymal stem cells (MSCs) in vitro and in vivo. In the in vitro studies, our results show that the weight ratio CSC:COL II that reaches 1.2:100 (CSC1.2/100 -COL II scaffold) can provide an optimal microenvironment for MSC chondrogenesis. When MSCs are cultured in this CSC1.2/100 -COL II scaffold, the chondrogenic gene expression of cultured cells is upregulated, while the osteogenic gene expression of these is downregulated. In addition, MSCs cultivated in the CSC1.2/100 -COL II scaffold are found to express the highest glycosaminoglycans:DNA ratio as compared to those in scaffolds of other CSC:COL II ratios. Histological and immunohistological evidence also supports the result. In the in vivo study, our results show that MSCs cultivated in the CSC1.2/100 -COL II scaffold demonstrate a better repair ability on cartilage lesions than does the COL II scaffold. After 1 month in vivo, the injected MSCs in the CSC1.2/100 -COL II scaffold show lacuna structures and stimulate the formation of type II collagen at the defective sites. Six months after transplantation, the generated cells in the CSC1.2/100 -COL II group show higher gene expressions of type II collagen and aggrecan but lower gene expression of type I collagen at the defective sites than those in the COL II group. The results strongly suggest that CSC1.2/100 -COL II scaffold can serve as a potential candidate for cartilage repair and improve the chondrogenesis of MSCs in general.


Assuntos
Condrogênese , Sulfatos de Condroitina/química , Colágeno Tipo II/química , Células-Tronco Mesenquimais/metabolismo , Alicerces Teciduais/química , Cartilagem/citologia , Cartilagem/metabolismo , Células Cultivadas , Colágeno Tipo I/biossíntese , Regulação da Expressão Gênica , Humanos , Células-Tronco Mesenquimais/citologia
17.
Toxicol Lett ; 212(1): 83-9, 2012 Jul 07.
Artigo em Inglês | MEDLINE | ID: mdl-22579953

RESUMO

RAD51 is essential for homologous recombination (HR)-mediated repair of DNA double-strand breaks (DSBs) in mammalian cells. RAD51 is an attractive target for anticancer drugs, given high RAD51 levels are frequently observed in many human tumors and associated with increased resistance to DSBs-inducing chemotherapeutics. Prodigiosin is a bacterial tripyrrole pigment with potent anticancer activity and also provokes DSBs. We hereby aimed to elucidate the role of RAD51 in prodigiosin-induced cytotoxicity. Prodigiosin was found to down-regulate RAD51 in multiple human breast carcinoma cell lines irrespective of p53 status. Mechanistically, prodigiosin lowered RAD51 mRNA expression, whereas blockade of proteasome-mediated degradation failed to restore RAD51 levels following prodigiosin treatment. In addition, prodigiosin triggered phosphorylation of JNK and p38 MAPK, while pharmacological inhibition of JNK or p38 MAPK attenuated prodigiosin-mediated inhibition of RAD51 mRNA expression. Lastly, cells with enforced RAD51 expression showed increased resistance to prodigiosin-induced cytotoxicity as well as inhibition of colony formation. Collectively, we conclude that RAD51 down-regulation represents one of the modes of prodigiosin's cytotoxic action, ostensibly by augmenting the genotoxic effect of prodigiosin through suppression of RAD51-mediated HR repair. Our findings further implicate the use of prodigiosin to potentiate the cytotoxicity of DSB-inducing chemotherapeutics through RAD51 down-regulation.


Assuntos
Antineoplásicos/farmacologia , Neoplasias da Mama/tratamento farmacológico , Prodigiosina/farmacologia , Rad51 Recombinase/metabolismo , Neoplasias da Mama/enzimologia , Neoplasias da Mama/metabolismo , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Feminino , Humanos , Immunoblotting , MAP Quinase Quinase 4/metabolismo , RNA Mensageiro/química , RNA Mensageiro/genética , Rad51 Recombinase/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
18.
J Biosci Bioeng ; 114(2): 198-203, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22578592

RESUMO

Lignocellulose was converted into reducing sugars by using saccharification enzymes from cocultivated Trichoderma reesei and Aspergillus niger and reducing sugars as nutrients for Zymomonas mobilis to produce bioethanol in an immobilization system. After 96 h of cultivation, cocultivated T. reesei and A. niger had enzymatical synergistic effects that enabled a reducing sugar production of 1.29 g/L and a cellulose conversion rate of 23.27%. An 18% total inoculum concentration and a 1/1 inoculation ratio of T. reesei to A. niger obtained a reducing sugar production rate and a cellulose conversion rate of 2.57 g/L and 46.27%, respectively. The co-immobilization cultivation results showed that using polyurethane as a carrier optimized total saccharification enzyme activity at an inoculum ratio of 1/1 and a total inoculum concentration of 6.5×10(6)spores/mL. Based on the experimental results, the bioreactor design was further modified to enhance bioethanol production. The three strains (A. niger, T. reesei and Z. mobilis) were cocultivated with a co-immobilization cultivation system. The experimental results showed that, after 24 h cultivation, bioethanol production reached 0.56 g/L, and reducing sugar conversion rate reached 11.2% when using carboxymethylcellulose (CMC) substrates. The experimental results confirmed that the modified bioreactor enhances bioethanol production. However, further experiments are needed to determine how to prevent multi-stage failure of reducing medium volume.


Assuntos
Reatores Biológicos , Celulose/metabolismo , Etanol/metabolismo , Alginatos , Aspergillus niger/enzimologia , Aspergillus niger/crescimento & desenvolvimento , Carboximetilcelulose Sódica/metabolismo , Células Imobilizadas/enzimologia , Células Imobilizadas/metabolismo , Celulose/química , Ácido Glucurônico , Ácidos Hexurônicos , Lignina/química , Lignina/metabolismo , Microesferas , Trichoderma/enzimologia , Trichoderma/crescimento & desenvolvimento , Zymomonas/crescimento & desenvolvimento , Zymomonas/metabolismo
19.
Appl Biochem Biotechnol ; 166(3): 796-804, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22203393

RESUMO

Cupriavidus necator is well known for its ability to accumulate polyhydroxybutyrate (PHB). When supplemented with propionic acid (or sodium propionate) in the growth medium, the bacterium is also able to synthesize polyhydroxybutyrate-co-hydroxyvalerate (PHBV). In order to increase the fraction of 3-hydroxyvalerate (3HV) in PHBV, we cloned the propionate permease gene prpP from C. necator and the propionyl-CoA synthase gene prpE from Cupriavidus taiwanensis and transformed into an Escherichia coli containing phaCAB operon of C. necator. The effects on PHBV accumulation in cells co-expressed with phaCAB and prpE or prpP in the media contained mixed carbon sources (glucose and sodium propionate) were evaluated. The HV fraction in PHBV increased when prpE or prpP was overexpressed in the cells. Concentrations of yeast extracts could also affect the fraction of HV. In addition, when glucose was replaced by sodium pyruvate, sodium succinate, or sodium gluconate, only PHB were detected in the recombinant strains.


Assuntos
Proteínas de Bactérias/genética , Coenzima A Ligases/genética , Cupriavidus necator/genética , Proteínas de Membrana Transportadoras/genética , Ácidos Pentanoicos/metabolismo , Proteínas de Bactérias/metabolismo , Clonagem Molecular , Coenzima A Ligases/metabolismo , Cupriavidus/genética , Cupriavidus/metabolismo , Cupriavidus necator/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Expressão Gênica , Glucose/metabolismo , Hidroxibutiratos/metabolismo , Proteínas de Membrana Transportadoras/metabolismo , Óperon , Poliésteres/metabolismo , Propionatos/metabolismo , Engenharia de Proteínas/métodos , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo
20.
Mar Drugs ; 9(4): 615-624, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21731553

RESUMO

Polyhydroxybutyrate (PHB) is one of the polyhydroxyalkanoates (PHAs) which has biodegradable and biocompatible properties. They are adopted in the biomedical field, in, for example, medical implants and drug delivery carriers. This study seeks to promote the production of PHB by Vibrio sp. BM-1, isolated from a marine environment by improving constituents of medium and implementing an appropriate fermentation strategy. This study successfully developed a glycerol-yeast extract-tryptone (GYT) medium that can facilitate the growth of Vibrio sp. BM-1 and lead to the production of 1.4 g/L PHB at 20 h cultivation. This study also shows that 1.57 g/L PHB concentration and 16% PHB content were achieved, respectively, when Vibrio sp. BM-1 was cultivated with MS-GYT medium (mineral salts-supplemented GYT medium) for 12 h. Both cell dry weight (CDW) and residual CDW remained constant at around 8.2 g/L and 8.0 g/L after the 12 h of cultivation, until the end of the experiment. However, both 16% of PHB content and 1.57 g/L of PHB production decreased rapidly to 3% and 0.25 g/L, respectively from 12 h of cultivation to 40 h of cultivation. The results suggest that the secretion of PHB depolymerase that might be caused by the addition of mineral salts reduced PHB after 12 h of cultivation. However, work will be done to explain the effect of adding mineral salts on the production of PHB by Vibrio sp. BM-1 in the near future.


Assuntos
Hidroxibutiratos/metabolismo , Poliésteres/metabolismo , Vibrio/metabolismo , Materiais Biocompatíveis/química , Materiais Biocompatíveis/metabolismo , Hidrolases de Éster Carboxílico/metabolismo , Meios de Cultura , Fermentação , Hidroxibutiratos/química , Minerais/química , Poliésteres/química , Sais/química , Fatores de Tempo , Vibrio/crescimento & desenvolvimento
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