Diabetic retinopathy (DR) can cause irreversible blindness and is the severest microvascular complication in the eyes of patients with diabetic mellitus (DM). The identification of susceptibility factors contributing to development of DR is helpful for identifying predisposed patients and improving treatment efficacy. Although proteomics analysis is useful for identifying protein markers related to diseases, it has never been used to explore DR-associated susceptibility factors in the aqueous humor (AH). To better understand the pathophysiology of DR and to identify DR-associated risk factors, a gel-based proteomics analysis was performed to compare AH protein profiles of DM patients with and without development of DR. MALDI-TOF MS was then performed to identify protein spots that were differentially expressed between the two groups and western blot analysis was used to validate the expressional change of protein demonstrated by proteomics. Our proteomics and bioinformatics analysis identified 11 proteins differentially expressed between DR and control groups. These proteins are linked to biological networks associated with nutrition transport, microstructure reorganization, angiogenesis, anti-oxidation, and neuroprotection. The data may provide potential AH biomarkers and susceptibility factors for predicting DR development, and provide an insight into the underlying pathophysiological mechanisms of DR. This article is part of a Special Issue entitled: Proteomics: The clinical link. 相似文献
We evaluated the presence of Ca, Na, K, Cu and Zn in the lenses and aqueous humour of rabbits treated with an Nd:YAG laser to induce opacity of the crystalline. The mean concentrations of the elements found in control lenses were: Ca: 15.8+/-5.2 mg/kg; Na: 1.2+/-0.6 g/kg; K: 10.3+/-3.3 g/kg; Cu: 0.19+/-0.06 mg/kg; Zn: 20.6+/-3.0 mg/kg. With the exception of K and Zn, the values found in the lenses of treated eyes (Ca: 135+/-24 mg/kg; Na: 4.3+/-1.5 g/kg; K: 10.1 +/- 3.2 g/kg; Cu: 0.47+/-0.17 mg/kg; Zn: 21.8+/-4.2 mg/kg) were significantly higher than in the controls. On the other hand, the concentrations found in the aqueous humour of treated eyes (Ca: 21.7+/-4.5 mg/l; Na: 0.66+/-0.21 g/l; K: 0.29+/-0.10 g/l; Cu: 0.035+/-0.009 mg/l; Zn: 0.079+/-0.01 mg/l) were significantly lower than those of the controls. The greatest difference was observed for Na (-68.6%) and Cu (-52.7%), followed by Ca and Zn (-35.0% and -35.2%, respectively). A positive correlation was found between Ca and Na in treated lenses (r2 = 0.9226, p < 0.0001) whereas inverse correlations were found for both Ca (r2 = 0.9788, p<0.0001) and Na (r2 = 0.9491, p<0.0001) between the concentrations found in the lenses and in the aqueous humour of treated eyes. 相似文献
Barley hull, a lignocellulosic biomass, was pretreated using aqueous ammonia, to be converted into ethanol. Barley hull was soaked in 15 and 30 wt.% aqueous ammonia at 30, 60, and 75 °C for between 12 h and 11 weeks. This pretreatment method has been known as “soaking in aqueous ammonia” (SAA). Among the tested conditions, the best pretreatment conditions observed were 75 °C, 48 h, 15 wt.% aqueous ammonia and 1:12 of solid:liquid ratio resulting in saccharification yields of 83% for glucan and 63% for xylan with 15 FPU/g-glucan enzyme loading. Pretreatment using 15 wt.% ammonia for 24–72 h at 75 °C removed 50–66% of the original lignin from the solids while it retained 65–76% of the xylan without any glucan loss.
Addition of xylanase along with cellulase resulted in synergetic effect on ethanol production in SSCF (simultaneous saccharification and co-fermentation) using SAA-treated barley hull and recombinant E. coli (KO11). With 3% w/v glucan loading and 4 mL of xylanase enzyme loadings, the SSCF of the SAA treated barley hull resulted 24.1 g/L ethanol concentration at 15 FPU cellulase/g-glucan loading, which corresponds to 89.4% of the maximum theoretical yield based on glucan and xylan.
SEM results indicated that SAA treatment increased surface area and the pore size. It is postulated that these physical changes enhance the enzymatic digestibility in the SAA treated barley hull. 相似文献
Carboxymethyl cellulase (CMCase) hydrolyses cellulose into glucose and is useful in various industrial applications. Conventional CMCase purification methods are rather complicated and time-consuming; thus, a cost-effective strategy for CMCase recovery is on demand. Polyethylene-glycol (PEG)/sodium citrate aqueous biphasic system (ABS) was adopted in this study to investigate the effectiveness of the ABS in the recovery of extracellular Bacillus subtilis CMCase from fermentation broth. Comprehensive optimization steps were executed that took into consideration the ABS variables of PEG molecular weight, tie-line length (TLL), volume ratio (VR), crude loading, pH and the addition of sodium chloride (NaCl). A CMCase recovery yield (YB) of 88.82% ± 0.69, a purification fold (PF) of 4.8 and a partition coefficient (K) of 0.44 ± 0.03 were achieved from the bottom phase of the PEG 6000/citrate ABS with TLL of 42.16% (w/w), VR of 0.29, 1% of (w/w) NaCl, pH 7.0, and 20% (w/w) crude loading. CMCase was mainly segregated to the salt-rich bottom phase because of the hydrophilicity of the enzyme surface. The highly effective recovery technique was further confirmed by SDS-PAGE analysis. Overall, the present study suggests that the ABS is a potential purification strategy for extracellular CMCase. 相似文献
The current optimization of IG-105 (3) on the carbazole-ring provided a series of new carbazole sulfonamides derivatives 13a–13m. All of the compounds have been evaluated against HepG2 cells (hepatoma cancer) for antiproliferative activity. Compounds that showed activity better or comparable to that of 3 versus HepG2 were evaluated against MCF-7 (breast cancer), MIA PaCa-2 (pancreatic cancer), and Bel-7402 (hepatoma/liver cancer) for antiproliferative activity. Of the seven compounds selected for further study five (13b, 13g, 13j, 13k and 13l) were found to give IC50 values against the four cell lines comparable to those for 3. Two compounds (13f and 13i) were more active than 3 and their activity against HepG2 and MCF-7 (IC50:0.01–0.07 μM) approached that of the positive controls podophyllotoxin (podo) and CA-4. Most of compounds showed aqueous solubility (0.11–19.60 μg/mL at pH 7.4 and 2.0) better than 3. These promising results warrant further development of new compounds 13f and 13i as potential potent antitumor drug candidates. 相似文献
This work describes the partition of a Schistosoma mansoni tegumental antigen produced by a recombinant Escherichia coli strain using an aqueous two-phase system (ATPS) composed of polyethylene glycol (PEG) and purified hydroxypropyl-starch (Reppal PES 100). The effects of the polymer molecular weight, tie line length and pH on antigen partitioning were investigated. The detection of the antigen in both phases was determined by ELISA. The system composed of PEG 8000 (5.1% w/w) and Reppal PES 100 (13.0% w/w) led to a yield of 92% and a purification factor of 12 concerning the antigen in the PEG-rich phase. It was observed that antigen partition in ATPSs was strongly affected by the pH and tie line length. In addition, it was possible in a single step, to remove the cell debris, which precipitated at the interface of the system. 相似文献
The effect of two inorganic salts, ammonium sulphate and potassium dihydrogenphosphate, on the partitioning of pectinases produced by Polyporus squamosus in polyethylene glycol/crude dextran aqueous two-phase system is reported. Presence of both salts at different concentration did not affect partition of biomass, so fungal growth was occurring exclusively in the bottom phase. At 30 mmol (NH4)2SO4/l in two-phase medium, the partition coefficient of endo-pectinase was 3.9, and it was 80% improved in comparison to that obtained at twofold lower salt concentration. On the other hand, higher (NH4)2SO4 concentration increased total exo-pectinase activity produced, but did not affect substantially its partition parameters. Increasing phosphate concentration stimulated partition of both enzymes to the top phase: at 0.2 mol KH2PO4/l the partition coefficient for exo-pectinase was about 20% higher than at 0.1 mol/l, and one-sided partition of endo-pectinase was accomplished, and consequently maximal top phase yield. 相似文献