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1.
Arch Med Res ; 50(6): 368-376, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31678895

RESUMO

BACKGROUND: This study aims to evaluate whether the Vitamin D receptor (VDR) gene polymorphisms were associated with systemic sclerosis (SSc) in a Chinese Han population. METHODS: Using a hospital-based case-control study including 100 SSc patients and 100 healthy controls. Single nucleotide polymorphisms (SNPs) in the VDR region were genotyped by the improved multiplex ligase detection reaction (i MLDR) method. Haplotypes were also constructed after linkage disequilibrium (LD) analysis. RESULTS: Eight SNPs (rs731236 (TaqI), rs2228570 (FokI), rs7975232 (ApaI), rs1544410 (BsmI), rs11574010 (Cdx2), rs739837 (BglI), rs757343 (Tru9I) and rs11168267) were included. There were significant differences between SSc patients and healthy individuals in ApaI and BglI genotype (both adjusted p = 0.008). Through the genotyping, significantly association of SSc were found for: dominant model of ApaI and BglI (both OR (95% CI) = 1.80 (1.03,3.16), p = 0.040). Furthermore, the elevation of erythrocyte sedimentation rate (ESR) had a higher percentage of BglI GT genotype frequency (p = 0.034) and dominant model of ApaI (p = 0.016) in SSc. There was high linkage disequilibrium was detected in BglI and ApaI polymorphisms (r2 = 1.0, D' = 1.0), Tru9I and rs11168267 (r2 = 0.926, D' = 0.969), respectively. No significant difference were found in these four haplotypes (all p >0.05). The correlation between VD levels and VDR gene polymorphisms was not statistically significant. CONCLUSIONS: Our preliminary study indicates the ApaI and BglI genotype may possibly have a role in the pathogenesis of SSc patients. Dominant model of ApaI and BglI GT genotype frequency may be associated with the increased risk of ESR.

2.
Int J Rheum Dis ; 22(10): 1832-1840, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31464381

RESUMO

AIM: To examine the associations between female menstrual or reproductive factors and the development of systemic sclerosis (SSc) in China. METHODS: In this hospital-based case-control study, for each subject, data on reproductive and menstrual factors such as number of births, abortions, and age at menarche were obtained by structured questionnaire. Risk estimates, measured by the odds ratio (OR) and 95% confidence interval (CI), were obtained by unconditional logistics regression. Furthermore, meta-analysis was performed and pooled OR with 95% CI for the number of pregnancies and abortions were calculated. RESULTS: There were 166 SSc and 392 female controls seen during the study period. The results showed women with late menarche age (≥17 years) were less likely than those with earlier age at menarche to develop SSc (OR 0.347, 95% CI 0.174-0.693) and compared with women without abortion, women with abortion (1 time) were at reduced risk of developing SSc (P = .036). After adjusting for potential confounders such as occupation and body mass index (BMI), late age at menarche (≥17 years) was associated with a decreased risk of SSc (OR 0.187, 95% CI 0.068-0.513), but abortions were not significantly related to SSc. The meta-analysis revealed there was no association between SSc and abortions or number of pregnancies. No significant publication bias was observed (P > .05). CONCLUSION: Late age at menarche was associated with a reduced risk of SSc but abortion may not be an independent risk factor for SSc. Further investigations are required to verify our findings.

3.
Autoimmunity ; 52(1): 21-26, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30822156

RESUMO

OBJECTIVES: Recent evidence has demonstrated that UBASH3A play a pivotal role in multiple autoimmune diseases. In this study, we explored the association between UBASH3A gene single-nucleotide polymorphisms (SNPs) and rheumatoid arthritis (RA) in a Chinese Han population. We also comparatively evaluated the UBASH3A expression profile in peripheral blood mononuclear cells (PBMCs) from patients with RA and healthy controls. METHODS: Four UBASH3A polymorphisms (rs1893592, rs11203203, rs2277798, and rs3788013) were studied in 553 patients with RA and 587 controls in a Chinese population. Genotyping was performed using the Fluidigm 192.24 Dynamic Array Integrated Fluidic Circuit (IFC). For gene expression study, UBASH3A mRNA levels of 30 RA patients and 31 healthy individuals were assessed by real-time quantitative polymerase chain reaction (RT-qPCR). Data were analyzed by SPSS 19.0 software. RESULTS: A significant association between rs1893592 polymorphism and RA was found under all genetic models (all p<.05). We also discovered a significant association between rs3788013 polymorphism and RA in the allele and genotype distributions, as well as the recessive model (all p<.05). Moreover, we found the genotype distribution and allele frequency of rs1893592 were significantly associated with RF phenotype in the RA patients (χ2 = 6.786, p=.034; χ2 = 4.534, p=.033; respectively). We also found the genotype distribution and allele frequency of rs2277798 were significantly associated with anti-CCP phenotype in the RA patients (χ2 = 7.873, p=.020; χ2 = 4.473, p=.034; respectively). However, we did not detect any significant associations between rs11203203 and RA susceptibility and autoantibody profiles (all p>.05). The mRNA expression of UBASH3A was increased in PBMCs of patients with RA when compared to healthy controls (p=.001). CONCLUSIONS: Our observations suggested that the dysregulation of UBASH3A might be associated with the pathogenesis of RA, and UBASH3A gene polymorphisms (rs1893592 and rs3788013) might contribute to RA susceptibility in Chinese Han population.

4.
Neurol Sci ; 39(2): 225-234, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29110148

RESUMO

Vitamin D receptor (VDR) polymorphisms have been inconsistently investigated in multiple sclerosis (MS). However, published studies demonstrated differences concerning design and effect size. A meta-analysis is necessary to determine the magnitude of the association between VDR polymorphisms and MS risk. The aim of the current study was to quantify the magnitude of the association between BsmI, FokI, ApaI, and TaqI VDR polymorphisms and MS risk. Following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, we conducted a systematic search and meta-analysis of the VDR gene polymorphisms and the risk of MS. The pooled odds ratios (OR) and 95% confidence interval (CI) were calculated by using Stata Version 11.0 with dominant and recessive models and allele analyses. A total of 4013 cases and 4218 controls in 24 case-control studies were included in the meta-analyses. The results did not indicate an association between any of the VDR polymorphisms and the risk of MS among overall populations, Asians, and Caucasians. However, our subgroup analysis suggests that the A allele was associated with MS risk in Asian populations (P = 0.005, OR = 1.267, 95% CI 1.074-1.496). Interestingly, the sensitivity analysis excluding studies with controls not in HWE showed insignificant association between the A allele and MS risk (P = 0.211), which was different from those in the non-sensitivity analysis. Our preliminary results indicate the VDR gene ApaI, BsmI, FokI, and TaqI polymorphisms may not be associated with elevated MS risk among overall populations. But ApaI polymorphism may confer different susceptibility to MS among different populations, and more well-designed studies with a large sample size are still needed to validate our results.


Assuntos
Predisposição Genética para Doença , Esclerose Múltipla/genética , Polimorfismo Genético/genética , Receptores de Calcitriol/genética , Grupo com Ancestrais do Continente Asiático/genética , Estudos de Casos e Controles , Humanos
5.
Int J Rheum Dis ; 20(12): 2053-2061, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28984075

RESUMO

OBJECTIVES: To identify the association of glucocorticoid receptor (GR) gene polymorphism with systemic lupus erythematosus (SLE). METHODS: A case-control study was carried out, in which 400 Chinese patients with SLE and 400 normal people were enrolled. DNA was extracted using a genomic DNA extraction kit, and tagged single nucleotide polymorphisms (SNPs) were identified by Haploview (4.0) Project from the Chinese HapMap Project. Eighteen tagged SNPs of the GR gene were genotyped by the Multiplex SNaPshot technique. RESULTS: Two GR gene SNPs were associated with the pathogenesis of SLE: rs6865292 (dominant model: crude odds ratio [OR] = 1.526, 95% CI: 1.151-2.025, P = 0.003; adjusted OR = 1.525, 95% CI: 1.149-2.023, P = 0.004; PBH = 0.036) and rs9324921 (dominant model: crude OR = 1.556, 95% CI: 1.173-2.062, P = 0.002; adjusted OR = 1.553, 95% CI: 1.171-2.060, P = 0.002; PBH = 0.036). The haplotype analysis of GR gene SNPs manifested that the haplotype of 'CCGGG' (OR = 2.701, 95% CI: 1.348-5.410, P = 0.004; PBH = 0.036) was a risk factor for the development of SLE. A lower frequency of A-allele of SNP rs4607376 (P = 0.021; OR = 0.794, 95% CI: 0.652-0.966, PBH = 0.126), higher frequency of C-allele of SNP rs6865292 (P = 0.019, OR = 1.317, PBH = 0.126) and A-allele of SNP rs9324921 (P = 0.019, OR = 1.317, PBH = 0.126) may be risk factors for developing SLE. The rs7719514 (recessive model: crude P = 0.044; adjusted P = 0.044, PBH = 0.264), rs7701443 (recessive model: crude P = 0.044, adjusted P = 0.045; PBH = 0.264), rs4607376 (recessive model: crude P = 0.027; adjusted P = 0.026; PBH = 0.264) and haplotype 'CAGCG' (P = 0.044; PBH = 0.198) showed marginal association with the pathogenesis of SLE. In the case group, there were no significant differences between non-lupus nephritis and lupus nephritis. Further, we found that the SNP rs12054797 (F = 3.228, P = 0.041, PBH = 0.342), rs2963156 (F = 3.163, P = 0.043, PBH = 0.342) might be marginally associated with disease activity. CONCLUSIONS: The study indicates that GR genetic polymorphisms may play a major role in the pathogenesis and development of SLE.


Assuntos
Lúpus Eritematoso Sistêmico/genética , Polimorfismo de Nucleotídeo Único , Receptores de Glucocorticoides/genética , Adulto , Grupo com Ancestrais do Continente Asiático/genética , Estudos de Casos e Controles , Distribuição de Qui-Quadrado , China/epidemiologia , Feminino , Frequência do Gene , Estudos de Associação Genética , Predisposição Genética para Doença , Haplótipos , Heterozigoto , Homozigoto , Humanos , Modelos Logísticos , Lúpus Eritematoso Sistêmico/diagnóstico , Lúpus Eritematoso Sistêmico/etnologia , Nefrite Lúpica/diagnóstico , Nefrite Lúpica/etnologia , Nefrite Lúpica/genética , Masculino , Pessoa de Meia-Idade , Razão de Chances , Fenótipo , Fatores de Risco , Adulto Jovem
6.
Mod Rheumatol ; 27(2): 298-305, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-27321124

RESUMO

BACKGROUND: Systemic sclerosis is an chronic inflammatory autoimmune diseases. Adipokine has been reported to play an important role in modulating immune responses. Recent studies suggest that adipokine also plays some roles in the pathogenesis of systemic sclerosis (SSc). However, published data regarding the relationship between plasma/serum adipokine levels and SSc are contradictory. The aim of this study was at performing a meta-analysis to derive a more accurate estimation and further investigate the association of plasma/serum leptin and adiponectin levels with SSc patients. METHODS: PubMed, and Web of Science databases (up to Feb 20, 2016) were used to obtain all relative published literatures. The study quality was assessed by the Newcastle-Ottawa scale. Pooled standard mean difference (SMD) with 95% confidence interval (CI) was calculated by random-effect model analysis. RESULTS: A total of fourteen studies were finally included in this meta-analysis. Among them, six of which were studied for the serum adiponectin levels in SSc patients, six of which were studied for the serum leptin levels in SSc patients, and two of them were studied both for serum adiponectin levels and serum leptin levels in SSc patients. The meta-analysis results showed that the serum adiponectin levels in SSc patients were significantly lower than that in normal controls (SMD = -0.608 ng/ml, 95% CI = -1.029 to -0.186, p = 0.005). However, there were no significant differences in serum leptin levels between SSc patients and healthy controls (SMD = -0.990 ng/ml, 95% CI = -2.340 to 0.359, p = 0.150). The subgroup analysis showed that Asia SSc patients with age less than 50 years old had lower plasma/serum adiponectin levels when compared with controls. CONCLUSION: The serum adiponectin levels, but not serum leptin levels, in SSc patients were significantly lower than that in normal controls.


Assuntos
Adiponectina/sangue , Leptina/sangue , Escleroderma Sistêmico/sangue , Adulto , Estudos de Casos e Controles , Humanos
7.
Breast ; 31: 214-218, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-27915203

RESUMO

PURPOSE: The assess the clinical value of different types of surgical procedures and further analyze the effect of adjuvant radiation therapy (RT) for adenoid cystic carcinoma (ACC) of the breast. METHODS: Patients with ACC of the breast were identified using a population-based national registration database (Surveillance, Epidemiology, and End Results, SEER). The Kaplan-Meier method and Cox regression models were performed to determine the impact of the surgical procedures and adjuvant RT associated with cause-specific survival (CSS) and overall survival (OS). RESULTS: A total of 478 patients with ACC of the breast were identified. The median follow-up was 59 months. The 10-year CSS and OS were 87.5% and 75.3%, respectively. For the Kaplan-Meier analysis, the 5-year CSS were 96.1%, 91.8%, 90.2%, and 94.1% in patients that received lumpectomy + adjuvant RT, lumpectomy alone, mastectomy alone, and mastectomy + adjuvant RT, respectively (p = 0.026). In the multivariate Cox analyses, lumpectomy + adjuvant RT was an independent prognostic factor for CSS and OS. Patients that received lumpectomy + adjuvant RT had better survival rates than patients that underwent lumpectomy only (CSS, p = 0.018; OS, p = 0.031) and mastectomy only (CSS, p = 0.010; OS, p = 0.004). CONCLUSION: ACC of the breast has an excellent prognosis. Breast-conserving surgery is a reasonable alternative for patients with ACC of the breast, and adjuvant RT after lumpectomy improved survival rates.


Assuntos
Neoplasias da Mama/patologia , Neoplasias da Mama/radioterapia , Carcinoma Adenoide Cístico/radioterapia , Carcinoma Adenoide Cístico/secundário , Mastectomia Segmentar , Neoplasias da Mama/cirurgia , Carcinoma Adenoide Cístico/cirurgia , Feminino , Seguimentos , Humanos , Estimativa de Kaplan-Meier , Metástase Linfática , Pessoa de Meia-Idade , Gradação de Tumores , Estadiamento de Neoplasias , Modelos de Riscos Proporcionais , Radioterapia Adjuvante , Programa de SEER , Taxa de Sobrevida , Carga Tumoral
8.
Onco Targets Ther ; 9: 67-73, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26730205

RESUMO

OBJECTIVE: To investigate the prognostic value of the lymph node ratio (LNR) in patients with small-cell carcinoma of the cervix (SCCC) after cancer-directed surgery using a population-based national registry (Surveillance Epidemiology and End Results [SEER]). METHODS: We retrospectively reviewed the data of SCCC patients in the SEER database from 1980 to 2012. The prognostic impact of LNR with respect to cause-specific survival (CSS) and overall survival (OS) was analyzed. RESULTS: A total of 118 patients with SCCC were identified. The median follow-up was 30.5 months. All these patients were treated with cancer-directed surgery and lymphadenectomy. Sixty (50.8%) patients had nodal metastases. The median LNR was 0.16 in patients with positive lymph nodes. Univariate analysis showed that prognostic factors such as International Federation of Gynecology and Obstetrics (FIGO) stage, nodal status, LNR, and local treatment modalities affected CSS and OS (P<0.05). Multivariate analysis showed that LNR was an independent prognostic factor for CSS and OS. Patients with a higher LNR had worse CSS (hazard ratio [HR]: 8.832; 95% confidence interval [CI]: 3.762-20.738; P<0.001) and OS (HR: 8.462; 95% CI: 3.613-19.821; P<0.001). LNR was associated with CSS and OS by stage, especially in FIGO stage I-II patients. CONCLUSION: LNR is an independent prognostic factor in SCCC patients and it may help to individualize adjuvant therapy.

9.
J Biosci Bioeng ; 118(1): 78-81, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24508023

RESUMO

D-P-Hydroxyphenylglycine (D-HPG) is a precursor required for the synthesis of semi-synthetic antibiotics. This unnatural amino acid can be produced by a transformation reaction mediated by D-hydantoinase (D-HDT) and d-amidohydrolase. In this study, a method was developed to integrate production and immobilization of recombinant D-HDT in vivo. This was approached by first fusion of the gene encoding D-HDT with phaP (encoding phasin) of Ralstonia eutropha H16. The fusion gene was then expressed in the Escherichia coli strain that carried a heterologous synthetic pathway for polyhydroxyalkanoate (PHA). As a result, d-HDT was found to associate with isolated PHA granules. Further characterization illustrated that D-HDT immobilized on PHA exhibited the maximum activity at pH 9 and 60°C and had a half-life of 95 h at 40°C. Moreover, PHA-bound d-HDT could be reused for 8 times with the conversion yield exceeding 90%. Overall, it illustrates the feasibility of this approach to facilitate in vivo immobilization of enzymes in heterologous E. coli strain, which may open a new avenue of enzyme application in industry.


Assuntos
Amidoidrolases/metabolismo , Escherichia coli/genética , Amidoidrolases/genética , Cupriavidus necator/genética , Enzimas Imobilizadas/genética , Enzimas Imobilizadas/metabolismo , Escherichia coli/metabolismo , Glicina/análogos & derivados , Glicina/metabolismo , Corpos de Inclusão , Lectinas de Plantas/genética , Poli-Hidroxialcanoatos/biossíntese
10.
Inflammation ; 36(6): 1424-30, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23839650

RESUMO

Although glucocorticoids (GCs) are effective in inducing remission in systemic lupus erythematosus (SLE) patients, there is a significant variation in response to therapeutic GCs, and some patients do not achieve full remission. The aim of this study was to explore the impact of environmental factors on the efficacy of GCs in a Chinese population with SLE. This was a prospective cohort study, and a total of 260 SLE patients treated with GCs (prednisone) were followed up for 12 weeks. The efficacy of GCs was measured with the scores on SLE disease activity index. Environmental factors were collected using a questionnaire. Single-variable analysis and multivariate logistic regression analysis were used to discriminate the impact of environmental factors on the efficacy of GCs. Two hundred forty-seven patients (95.00 %) completed the 12-week follow-up. Among these patients, 131 (53.04 %) were classified into sensitive group and 116 (46.96 %) were classified into insensitive group. Results from logistic analysis showed that the following environmental factors were significantly associated with decreased efficacy of GCs: high salt intake (OR = 3.464, 95%CI = 1.481-8.102, P = 0.004), introverted personality (OR = 3.550, 95%CI = 1.901-6.628, P < 0.0001), experience with negative life events (OR = 5.526, 95%CI = 1.612-18.946, P = 0.007), and history of allergy (OR = 2.966, 95%CI = 1.312-6.704, P = 0.009). These results indicate that environmental factors, including salt intake, personality, experience with negative life events, and history of allergy, may play an important role in the efficacy of GCs in the Chinese population with SLE.


Assuntos
Exposição Ambiental/efeitos adversos , Glucocorticoides/uso terapêutico , Lúpus Eritematoso Sistêmico/tratamento farmacológico , Prednisona/uso terapêutico , Adulto , China , Estudos de Coortes , Feminino , Humanos , Hipersensibilidade , Acontecimentos que Mudam a Vida , Masculino , Personalidade , Estudos Prospectivos , Cloreto de Sódio , Inquéritos e Questionários
11.
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
12.
Methods Mol Biol ; 765: 113-23, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21815090

RESUMO

Genetic manipulation of Escherichia coli strains for desired traits is the most applied strain engineering approach in industrial applications. For chromosomal insertion of genes and controlled expression of genomic genes in E. coli, the replicon-free and markerless method is described based on a series of conditional-replication plasmids called phage-integration vectors. They mainly carry the multiple cloning site and the prophage attachment site, which are sandwiched by two FRT sites. With the aid of the phage integrase from conditional-replication helper plasmids, the passenger genes of either foreign or native type incorporated into the integration vectors can be specifically integrated into bacterial genome at the prophage attachment site. Finally, the inserted DNA containing replicon and/or selective markers in integrants can be eliminated by the act of the FLP recombinase provided from a conditional-replication helper plasmid.


Assuntos
Cromossomos Bacterianos/genética , Escherichia coli/genética , Expressão Gênica , Genes/genética , Engenharia Genética/métodos , Vetores Genéticos/genética , Mutagênese Insercional/genética , Genes Bacterianos/genética , Genoma Bacteriano/genética
13.
Biotechnol Bioeng ; 108(7): 1662-72, 2011 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-21337329

RESUMO

Targeting of non-phagocytic tumor cells and prompt release of gene cargos upon entry into tumors are two limiting steps in the bacterial gene delivery path. To tackle these problems, the non-pathogenic Escherichia coli strain BL21(DE3) was engineered to display the anti-HER2/neu affibody on the surface. After co-incubation with tumor cells for 3 h, the anti-HER2/neu affibody-presenting E. coli strain was selectively internalized into HER2/neu-positive SKBR-3 cells. The invasion efficiency reached as high as 30%. Furthermore, the bacteria were equipped with the phage ϕX174 lysin gene E-mediated autolysis system. Carrying the transgene (e.g., eukaryotic green fluorescent protein, GFP), the tumor-targeting bacteria were subjected to the thermal shock to trigger the autolysis system upon entry into HER2/neu-positive cells. Flow cytometric analysis revealed that 3% of infected cells expressed GFP 24 h post thermal induction. Overall, the results show a promise of the proposed approach for developing bacteria as a delivery carrier.


Assuntos
Escherichia coli/genética , Escherichia coli/fisiologia , Técnicas de Transferência de Genes , Transferência Genética Horizontal , Transgenes , Anticorpos/imunologia , Anticorpos/metabolismo , Adesão Celular , Linhagem Celular Tumoral , Humanos , Ligação Proteica , Receptor ErbB-2/imunologia , Receptor ErbB-2/metabolismo
14.
J Biosci Bioeng ; 111(3): 336-42, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21159550

RESUMO

A halophilic bacterium isolated from a salt environment in southern Taiwan was identified as a Marinococcus sp. ECT1. This bacterium could synthesize and accumulate intracellular ectoine as a compatible solute capable of resisting osmotic stress in a hyper-osmotic environment. This study also developed a semi-synthesized medium (YAMS medium), capable of facilitating the growth of this Marinococcus sp. ECT1 with 600 mg/L crude ectoine production. Moreover, Marinococcus sp. ECT1 was grown on YAMS medium containing different initial yeast extract concentrations (C(YE)) (0 to 60 g/L) to demonstrate how C(YE) affects crude ectoine production. While the maximum cell concentration was increased by 23-fold when the C(YE) was 40 g/L, the maximum crude ectoine production reached 2.5 g/L when C(YE) was 40 g/L. In addition to demonstrating the success of the fermentation strategy of ectoine in increasing the production and production yield, experimental results further demonstrated that the fermentation medium of ectoine is highly promising for commercialization. Furthermore, the molecular weight and chemical structure of ectoine were identified and characterized by FAB-MS and (1)H-NMR.


Assuntos
Diamino Aminoácidos/biossíntese , Bacillaceae/metabolismo , Fermentação , Microbiologia Industrial , Bacillaceae/crescimento & desenvolvimento , Carbono/metabolismo , Meios de Cultura/química , Nitrogênio/metabolismo , Salinidade , Cloreto de Sódio/química , Taiwan
15.
Int J Mol Sci ; 11(11): 4526-38, 2010 Nov 12.
Artigo em Inglês | MEDLINE | ID: mdl-21151454

RESUMO

Fengycin, a lipopeptide biosurfactant, was produced by indigenous Bacillus subtilis F29-3 isolated from a potato farm. Although inhibiting the growth of filamentous fungi, the fengycin is ineffective against yeast and bacteria. In this study, fengycin was isolated from fermentation broth of B. subtilis F29-3 via acidic precipitation (pH 2.0 with 5 N HCl) followed by purification using ultrafiltration and nanofiltration. The purified fengycin product was characterized qualitatively by using fast atom bombardment-mass spectrometer, Fourier transform infrared spectrometer, ultraviolet-visible spectrophotometer, (13)C-nuclear magnetic resonance spectrometer and matrix assisted laser desorption ionization-time of flight, followed by quantitative analysis using reversed-phase HPLC system. This study also attempted to increase fengycin production by B. subtilis F29-3 in order to optimize the fermentation medium constituents. The fermentation medium composition was optimized using response surface methodology (RSM) to increase fengycin production from B. subtilis F29-3. According to results of the five-level four-factor central composite design, the composition of soybean meal, NaNO(3), MnSO(4)·4H(2)O, mannitol-mannitol, soybean meal-mannitol, soybean meal-soybean meal, NaNO(3)-NaNO(3) and MnSO(4)·4H(2)O-MnSO(4)·4H(2)O significantly affected production. The simulation model produced a coefficient of determination (R(2)) of 0.9043, capable of accounting for 90.43% variability of the data. Results of the steepest ascent and central composite design indicated that 26.2 g/L of mannitol, 21.9 g/L of soybean meal, 3.1 g/L of NaNO(3) and 0.2 g/L of MnSO(4)·4H(2)O represented the optimal medium composition, leading to the highest production of fengycin. Furthermore, the optimization strategy increased the fengycin production from 1.2 g/L to 3.5 g/L.


Assuntos
Bacillus subtilis/metabolismo , Lipopeptídeos/química , Bacillus subtilis/crescimento & desenvolvimento , Biotecnologia/métodos , Meios de Cultura/química , Fermentação , Lipopeptídeos/biossíntese
16.
Appl Biochem Biotechnol ; 162(8): 2355-64, 2010 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-20556541

RESUMO

Polyhydroxyalkanoates are polyesters synthesized by numerous microorganisms. These polyesters are biodegradable and have similar properties to those of conventional plastics. Cupriavidus taiwanensis strain 184 is phylogenetically related to the well-known polyhydroxybutyrate (PHB) producer Ralstonia eutropha (Cupriavidus necator) and is also shown to be able to accumulate significant amounts of PHB. In this study, we cloned the PHB synthesis genes (phaCAB) from C. taiwanensis 184 into Escherichia coli for biosynthesis of PHB. The recombinant E. coli strains were able to synthesize significant amounts of PHB. The PHB amounted to about 66~70% of total cell material of these recombinant strains.


Assuntos
Cupriavidus/genética , Escherichia coli/genética , Genes Bacterianos/genética , Poliésteres/metabolismo , Clonagem Molecular , DNA Recombinante/genética , Expressão Gênica , Dados de Sequência Molecular , Óperon/genética , Poliésteres/isolamento & purificação , Reação em Cadeia da Polimerase
17.
Zhonghua Nei Ke Za Zhi ; 48(2): 136-9, 2009 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-19549470

RESUMO

OBJECTIVE: To characterize the clinical characteristics of lupus mesenteric vasculitis (LMV). METHODS: Analyzing the clinical, laboratory and treatment data of LMV patients hospitalized from 2002.1.1 to 2007.12.31 retrospectively. RESULTS: (1) The three common manifestations were abdominal pain, diarrhea and vomit with the prevalence rate of 77%, 70% and 67% respectively. (2) The majority of LMV cases were active vital organ (28/30), kidney (24/30) and hematological system (18/30) were the main organs of involvement. Ten patients had hydroureteronephrosis, and 8 patients had intestinal pseudo-obstruction at the same time. (3) Systemic lupus erythematosus disease activity index (SLEDAI) score was > or = 10 in 80% (24/30) of patients. The progression of LMV was accompanied with new-onset leucopenia or worsening leucopenia or hypocomplementemia in 10 cases. (4) Blood antinuclear antibodies were positive in 27 patients detected, and anti-SSA antibody was positive in 15 (56%), anti-U(1)RNP antibody was positive in 14 (52%). (5) Fourteen cases had bowel wall thickening with target sign or mesenteric vessels with palisade or comb sign in contrast CT scan of abdomen. (6) Twenty-seven cases were treated with orally or intravenous medium to high dose steroid therapy and recovered from LMV. CONCLUSIONS: (1) Abdominal pain, diarrhea and vomit were frequent manifestations of LMV patients. (2) LMV was one of the serious complications of systemic lupus erythematosus (SLE), and usually accompanied by active SLE in other organs. (3) A drop in the white blood cell count or complement C(3) titer might be correlate with the occurrence of LMV. It needs to further investigate the relationship between LMV and the high positive rate of anti-SSA and anti-U(1)RNP antibody. (4) LMV patients responded well to intravenous high dose methylprednisolone.


Assuntos
Gastroenterite/etiologia , Lúpus Eritematoso Sistêmico/complicações , Vasculite/etiologia , Adolescente , Adulto , Feminino , Gastroenterite/diagnóstico , Gastroenterite/tratamento farmacológico , Glucocorticoides/uso terapêutico , Humanos , Lúpus Eritematoso Sistêmico/diagnóstico , Lúpus Eritematoso Sistêmico/tratamento farmacológico , Masculino , Artérias Mesentéricas/patologia , Veias Mesentéricas/patologia , Mesentério/irrigação sanguínea , Pessoa de Meia-Idade , Estudos Retrospectivos , Vasculite/diagnóstico , Vasculite/tratamento farmacológico , Adulto Jovem
18.
Appl Microbiol Biotechnol ; 76(1): 67-74, 2007 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-17457541

RESUMO

Rhamnolipid is one of the most commonly used biosurfactants with the ability to reduce the surface tension of water from 72 to 30 mN/m. An indigenous isolate Pseudomonas aeruginosa S2 possessing excellent ability to produce rhamnolipid was used as a model strain to explore fermentation technology for rhamnolipid production. Using optimal medium and operating conditions (37 degrees C, pH 6.8, and 250 rpm agitation) obtained from batch fermentation, P. aeruginosa S2 was able to produce up to 5.31 g/l of rhamnolipid from glucose-based medium. To further improve the rhamnolipid yield, a pH-stat fed-batch culture was performed by maintaining a constant pH of 6.8 through manipulating glucose feeding. The effect of influent glucose concentration on rhamnolipid yield and productivity was investigated. Using the pH-stat culture, a maximum rhamnolipid concentration (6.06 g/l) and production rate (172.5 ml/h/l) was obtained with 6% glucose in the feed. Moreover, combining pH-stat culture with fill-and-draw operation allowed a stable repeated fed-batch operation for approximately 500 h. A marked increase in rhamnolipid production was achieved, leading to the best rhamnolipid yield of approximately 9.4 g/l during the second repeated run.


Assuntos
Glicolipídeos/biossíntese , Pseudomonas aeruginosa/metabolismo , Tensoativos/metabolismo , Meios de Cultura , Fermentação , Glucose , Concentração de Íons de Hidrogênio , Pseudomonas aeruginosa/crescimento & desenvolvimento
19.
Biotechnol Prog ; 23(3): 661-6, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-17461551

RESUMO

An indigenous strain Pseudomonas aeruginosa S2 (P. aeruginosa S2), isolated from diesel-contaminated soil, produced extracellular surface-active material identified as rhamnolipid. Due to its excellent surface activity, rhamnolipid is known to be well-suited for stimulating the bioremediation efficiency of oil contaminated sites. To improve production yield of rhamnolipid with P. aeruginosa S2, various carbon and nitrogen sources were screened to select favorable ones leading to better biosurfactant production yield. It was found that using 4% glucose could attain better rhamnolipid yield, while 50 mM NH4NO3 appeared to be the most preferable nitrogen source. Meanwhile, the effect of carbon to nitrogen ratio (C/N ratio) on rhamnolipid yield was also investigated, and the optimal C/N ratio was identified as approximately 11.4. Moreover, response surface methodology (RSM) was applied to optimize the trace element concentration for rhamnolipid production. Results from two-level design indicate that concentrations of MgSO4 and FeSO4 were the most significant factors affecting rhamnolipid production. Using steepest ascent method and RSM analysis, an optimal medium composition was determined, giving a rhamnolipid production yield of 2.37 g/L in 100 h at 37 degrees C and 200 rpm agitation. Scale-up production of rhamnolipid in a well-controlled 5 L jar fermentor using the optimal medium and operating condition (at 37 degrees C and pH 6.8) further elevated the biosurfactant production yield to 5.31 g/L (in 97 h), which is over 2-fold higher than the best results obtained from shake-flask tests.


Assuntos
Glicolipídeos/metabolismo , Pseudomonas aeruginosa/metabolismo , Tensoativos/metabolismo , Carbono/farmacologia , Cromatografia Líquida de Alta Pressão , Meios de Cultura/farmacologia , Fermentação/efeitos dos fármacos , Espectrometria de Massas , Nitrogênio/farmacologia , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/crescimento & desenvolvimento
20.
Biotechnol Lett ; 26(10): 799-802, 2004 May.
Artigo em Inglês | MEDLINE | ID: mdl-15269550

RESUMO

Serratia marcescens SS-1 and its SpnR-defective isogenic mutant, SMdeltaR, produced an extracellular surfactant able to decrease surface tension of water from 72 to 37 dyne cm(-1) (SMdeltaR strain) and to 45 dyne cm(-1) (SS-1 strain). The biosurfactant also emulsified kerosene and diesel with a maximum emulsion index of 77% (diesel and kerosene) for the SMdeltaR strain, and 72% (kerosene) and 40% (diesel) for the SS-1 strain. Deletion of spnR gene appeared to enhance biosurfactant production. Model simulations suggest that biosurfactant production by the two strains was growth-associated. The SMdeltaR strain had a yield coefficient of 22-32% g dry cell(-1), which is 32-50% higher than that of the SS-1 strain.


Assuntos
Serratia marcescens/crescimento & desenvolvimento , Serratia marcescens/metabolismo , Tensoativos/química , Tensoativos/metabolismo , Transativadores/deficiência , Emulsões/química , Gasolina , Engenharia Genética/métodos , Querosene , Proteínas Repressoras , Serratia marcescens/classificação , Especificidade da Espécie , Tensão Superficial
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