Berberine and itraconazole are not synergistic in vitro against Aspergillus fumigatus isolated from clinical patients

The incidence of Aspergillus fumigatus infections has become more frequent as a consequence of widespread immunosuppression. At present, the number of available antifungal agents in the clinic is limited, and most of them, such as itraconazole (ICZ), are toxic and show resistance. Berberine (BER) is a plant alkaloid used in the clinic mainly for alimentary infections. We have used BER and ICZ to measure in vitro resistance in A. fumigatus isolated from clinical patients. The minimum inhibitory concentration ranges of BER and ICZ were 4-256 and 0.031-0.250 μg/mL, respectively. In addition, against A. fumigatus IFM 40808 strain, the MIC?? values of BER and ICZ were 8 and 0.125 μg/mL. Using this strain, we compared the giant colonies with or without BER, and concluded that BER could restrain A. fumigatus mycelial growth and conidial pigment production. Combinations of the two drugs were also tested by the checkerboard assay to identify any functional interactions between them. Thirty-two out of 42 isolates had FICI values > 4.0, indicating that two drugs were mutually antagonistic. In conclusion, it is not advised that BER and ICZ be used in the clinic at the same time. Our results indicated that BER may inhibit A. fumigatus through the ergosterol biosynthesis pathway, like ICZ.     

Purification and characterization of an exoinulinase from Aspergillus fumigatus

An extracellular exoinulinase was purified from the crude extract of Aspergillus fumigatus by ammonium sulfate precipitation, followed by successive chromatographies on DEAE-Sephacel, Sephacryl S-200, concanavalin A-linked amino-activated silica, and Sepharose 6B columns. The enzyme was purified 25-fold, and the specific activity of the purified enzyme was 171 IU/mg of protein. Gel filtration chromatography revealed a molecular weight of about 200 kDa, and native polyacrylamide gel electrophoresis (PAGE) showed an electrophoretic mobility corresponding to a molecular weight of about 176.5 kDa. Sodium dodecyl sulfate-PAGE analysis revealed three closely moving bands of about 66, 62.7, and 59.4 kDa, thus indicating the heterotrimeric nature of this enzyme. The purified enzyme appeared as a single band on isoelectric focusing, with a pI of about 8.8. The enzyme activity was maximum at pH 5.5 and was stable over a pH range of 4.0–9.5, and the optimum temperature for enzyme activity was 60°C. The purified enzyme retained 35.9 and 25.8% activities after 4 h at 50 and 55°C, respectively. The inulin hydrolysis activity was completely abolished with 1 mM Hg⁺⁺, whereas EDTA inhibited about 63% activity. As compared to sucrose, stachyose, and raffinose, the purified enzyme had lower K _m (0.25 mM) and higher V _max (333.3 IU/mg) values for inulin.     

Effects of gamma irradiation on solid and lyophilised phospholipids

The effects of gamma irradiation (25 kGy) as a sterilisation method for phospholipids (distearoylphosphatidylcholine and distearoylphosphatidylglycerol) were investigated. ³¹P-NMR revealed minor chemical degradation of the phospholipids but lower dynamic viscosity and pseudoplasticity, lower turbidity, higher diffusion constant, smaller size, more negative zeta potential and changes in the phase transition behaviour of the subsequently produced liposomes were observed. The observed changes could to some extent be explained by the irradiation-induced degradation products (distearoylphosphatidic acid, fatty acids, lysophospholipids).     

Effects of gamma and electron beam irradiation on polymer electrolytes

Polymer electrolytes based on poly(ethylene oxide) and lithium salts have been widely studied in recent years. In order to enhance the room temperature ionic conductivity of PEO-LiX complexes, various techniques, such as addition of plasticizers and crown ether, and also irradiation by γ and electron beams have been investigated. The enhancement of the conductivity by irradiation has been accounted for the decreasing of the crystallinity of PEO-LiX. We reviewed these results and have investigated the degradation processes of PEO using Tb³⁺ fluorescence probes. We have also studied on the effects of irradiation of polymers such as poly(acrylic acid) (PAA), poly(methacrylic acid) (PMA) and PEO using Tb⁺³ fluorescence probe. Various monomers containing SO₃H and COOH have been grafted on poly(ethylene oxide) using irradiation technique. The structures and ionic conductivities of Li and Na salts of irradiated products were investigated in detail.     

Effects of aeration on gamma irradiation of sewage sludge

In this paper the effect of aeration on gamma irradiation of sewage sludge was investigated to examine the potential solubilization of solids in sewage sludge to ultimately reduce the solids volume for disposal. Results showed that aeration increased the effectiveness of gamma radiation. The efficiency of sludge solubilization with aeration was increased by around 25% compared to that without aeration at an irradiation dose of 2.5–9 kGy. The soluble protein, polysaccharide and humic (like) substance concentrations were higher under aerated conditions. With aeration the overall reaction appears to be oxidative as evidenced by the higher nitrate and nitrite ion concentrations in solution.     

Expression and Characterization of a Novel Enantioselective Lipase from Aspergillus fumigatus

A 1,080-bp cDNA (CGMCC 2873) encoding of a cold-active lipase of Aspergillus fumigatus (AFL67) was cloned and expressed in Escherichia coli for the first time. The new lipase, AFL67, was one-step purified by 8.30 folds through Ni?CNTA affinity chromatography with a recovery of 86.8?%. The specific activity of purified AFL67 was 449?U?mg^?1 on p-NP hexanoate. AFL67 preferentially hydrolyzed p-nitrophenyl esters of short- and medium-chain fatty acids, with p-nitrophenyl hexanoate the maximum. The optimum temperature and pH was 15?°C and 7.5, respectively. The purified AFL67 was stable at 10?C25?°C for 30?min, and in the pH range of 6.0?C9.0 for 16?h (at 4?°C). Its activity was increased by 47 and 50?%, in the presence of 10?% (v/v) ethanol and isopropanol, respectively. The new lipase AFL67 highly enantioselectively deacylated (S)-??-acetoxyphenylacetic acid (APA) and o-Cl-APA, m-Cl-APA, and p-Cl-APA to (S)-mandelic acid and its derivates. These features render this cold-active novel lipase AFL67 attractive for biotechnological applications in the field of enantioselective synthesis of chiral mandelic acids, o-acylated mandelic acids, and their derivates and detergent additives.     

Effects of aromatic diacetylenes on polyurethane degradation by gamma irradiation

The effects of four aromatic diacetylenes on the gamma-irradiation-induced degradation of a commercial polyurethane were studied. Addition of 1 wt% of diphenylbutadiyne, which is homogeneously distributed in the polymer, effectively suppressed the polymer chain degradation. The dose required to decrease tensile strength by 50% was found to be 582 kGy for the polyurethane with 1 wt% of diphenylbutadiyne, while a dose of only 310 kGy for the polyurethane itself. The Young's modulus of the polyurethane alone decreased with dose; meanwhile the films with diphenylbutadiyne did not change. The films with p,p′-dinitrodiphenylbutadiyne behaved differently from others due to their inhomogeneous composition. The amide substituted diacetylenes also showed protecting effect, but in less extent due to the steric effect.     

Effects of gamma irradiation on poly(ethylene isophthalate)

Radiation methods are largely used for polymerisation and polymer modification, since irradiation induces transformations in the structure of materials which can be exploited to improve their performance. On the other hand, combined action of ionising radiation and oxygen may lead to degradation of the polymer, with worsening of properties such as mechanical strength or electrical insulation resistance. Therefore, the change of the chemical and physical properties of polymers under irradiation is a dynamic topic of research. In this work there are discussed data on the physical features of a polyester, poly(ethylene isophthalate) (PEI), subjected to gamma irradiation up to 1 MGy. PEI is a semicrystalline polymer with a structure similar to polyethylene terephthalate. Viscosity and differential scanning calorimetry measurements were carried out which allowed the monitoring of changes in the structure in terms of variations in the molecular weight, as well as of the percentage crystallinity depending on the dose. Furthermore, positron annihilation lifetime spectroscopy supplied information on the free volume present in the amorphous phase of the irradiated polymer.     

Effects of gamma irradiation on the single crystal ergosterol: An EPR study

Single crystals of ergosterol were investigated by Electron Paramagnetic Resonance (EPR) technique, with γ irradiation of the crystals at different orientations in the magnetic field between temperatures of 120 and 380 K, and the spectra were found to be slightly dependent on temperature. Because of the importance of ergosterol it is important to determine the irradiation effects on this molecule.Taking into consideration the chemical structure and the experimental spectra of the irradiated single crystal ergosterol, we found that two paramagnetic species which were labeled as radical A, CH_2αH_β, and radical B, CH_αH_βH_γH_σ, were produced in the host crystal. The EPR parameters; spectroscopic splitting factor, g, and hyperfine coupling constant, a, were determined for each radical.     

Effects of storage and gamma irradiation on (japonica) waxy rice

Japonica cultivar, Taichung waxy 70 (TCW 70), was exposed to gamma radiation with doses ranging from 0 to 2.0 kGy. The effects of gamma irradiation on waxy rice pasting properties and the qualities of its food product, mochi, were compared to the effects of storage over 12 months. Doses ranging from 0.5 to 2 kGy can decrease the paste viscosity of waxy rice as those obtained after 6 months of storage. Radiation treatments were less effective to decrease the hardness of mochi than waxy rice samples stored over 6 months. It was shown the effects of gamma irradiation on shortening the japonica waxy rice aging time and improving the quality of rice products, like mochi, were not as good as the effects of storage.     

Effects of gamma irradiation and accelerated aging on GO/UHMWPE nanocomposites

This article investigates irradiated and accelerated aged graphene oxide (GO)/ultrahigh molecular weight polyethylene (UHMWPE) nanocomposites. The prepared GO/UHMWPE nanocomposites are gamma-irradiated at a high irradiation dose in a vacuum and then accelerated aging procedure is performed at 80°C in an air oven for 21 days. Irradiated and aged samples are characterized by Raman spectrum, Fourier transform infrared (FT-IR) spectrum, differential scanning calorimetry, contact angle, and gel content. Filling GO reduces the intensity of Raman spectrum of UHMWPE and irradiation or aging cannot affect vibrational modes of UHMWPE and GO/UHMWPE. The result of the FT-IR spectrum shows that UHMWPE and GO/UHMWPE basically have the same oxidation index values, whether with irradiation or accelerated aging. Irradiation or aging can slightly increase the melting temperature. GO, irradiation, or aging can significantly increase the crystallinity and improve wetting properties. In irradiated GO/UHMWPE, GO is able to maintain the efficiency of the cross-linking. However, after aging, the cross-linking density of GO/UHMWPE is reduced significantly. According to the above results, it is proposed that GO shows a very weak scavenging free radicals capacity in GO/UHMWPE composites and cannot display antioxidant capacity.     

Effects of gamma irradiation on cell-wall constituents of some agricultural residues

The effects of 150 kilogray (kGy) of γ irradiation on cell-wall constituents of cottonwood (CW), lentils straw (LS), apple pruning products (AP) and olive cake (OC) were investigated. Samples were irradiated by γ irradiation at a dose level of 150 kGy under identical conditions of temperature and humidity and analyzed for crude fibre (CF), neutral-detergent fibre (NDF), acid detergent fibre (ADF) and acid-detergent lignin (ADL). The results indicate that γ irradiation decreased CF contents by about 29% for CW, LS and AP and by 17% for OC. NDF values were also decreased by about 4% for CW and OC, and by about 12% for LS and AP. γ Irradiation treatment also decreased ADF values only for CW by 8%. ADL contents decreased by 8% for CW and 5% for OC with no effects for LS and AP. The percentage of cellulose (CL): CF ratio increased by 30, 34, 38 and 20% for CW, LS, AP and OC, respectively. Also, the percentage of hemicellulose (HCL): CF increased by 57% for CW and 16% for OC and decreased by 7% for LS and AP. The percentage of HCL: ADL increased by 22% for CW but decreased by 33% for LS and AP with no changes for OC. There were no changes in CL: ADL ratio for all residues.     

Effects of gamma irradiation on chromatophores and volatile components of grass shrimp muscle

Effects of medium-dose (5 kGy) irradiation on chromatophores and volatile components of shrimp muscle were studied by light microscope and gas chromatography. Medium-dose irradiation treatment can inhibit the dispersion of erythrin in chromatophores but not the dispersion of melanins. No significant change was found in volatile composition of shrimp after irradiation.     

Effects of gamma irradiation on the physical and structural properties of β-glucan

This study was carried out to evaluate the effect of gamma irradiation on the physical and structural properties of β-glucan. β-Glucan solution (10%, w/v) was exposed to a cobalt-60 source (10, 30, and 50 kGy). Gel permeation chromatography data showed that the average molecular weight of irradiated β-glucan significantly decreased as the irradiation dose increased. In addition, gamma irradiation improved the solubility and decreased the viscosity of β-glucan by the radiolysis of the glycosidic bonds, and this effect was dependent upon the absorbed dose. Fourier transform infrared spectroscopy results showed that the functional groups of β-glucan were not significantly affected by gamma irradiation. Scanning electron microscopy results showed that the irradiated β-glucan was deformed into smaller granules. Therefore, gamma irradiation could be used in commercial processes as an effective method to resolve the physical problems involved in the use of β-glucan with high viscosity and low solubility.     

Expression and Characterization of a Novel Lipase from Aspergillus fumigatus with High Specific Activity

A novel lipase gene from Aspergillus fumigatus, afl1-1, was cloned and expressed with a molecular mass of 38 kDa in Escherichia coli for the first time. The recombinant lipase had a preference for short carbon chain p-nitrophenyl esters, especially toward C2 p-nitrophenyl ester and exhibited potent hydrolysis activity that had not been observed. The optimum pH and temperature of this new enzyme were 8.5 and 65 °C, respectively. The recombinant lipase (AFL1-1) is an alkaline enzyme which was stable in the pH range 6.0～8.5 for 16 h (at 4 °C) and at 30～50 °C for 1 h. It is an intracellular enzyme which was purified approximately 8.47-fold with an overall yield of 86.1% by single-step Ni-NTA affinity purification, with a very high specific activity of approximately 1.00?×?10(3) U mg(-1) on a standard substrate of p-nitrophenyl acetate. The Michaelis-Menten kinetic parameters V (max) and K (m) of the lipase were 1.37 mM mg(-1) min(-1) and 14.0 mM, respectively. Ca(2+) and other metal ions could not activate the lipase. According to the homology analysis and site-directed mutagenesis assay, the catalytic triad of the recombinant lipase was identified as Ser-165, Asp-260, and His-290 residues.     

Effects of gamma irradiation on chemical,microbial quality and shelf life of shrimp

In the present study the combined effect of gamma irradiation (1, 3 and 5 kGy) and storage at two temperatures: refrigeration (+4 °C) and frozen (?18 °C), on the shelf-life extension of fresh shrimp meat was investigated. The study was based on microbiological and physicochemical changes occuring in the shrimp samples. Total volatile base nitrogen values and trimethylamine values for irradiated shrimp samples were significantly lower than non-irradiated samples at both storage temperatures, and the rate of decrease was more pronounced in samples irradiated at the higher dose (p<0.05). Thiobarbituric acid values for irradiated shrimp samples were significantly higher than non-irradiated samples at both storage temperatures (p<0.05). pH values of shrimp samples were affected significantly by both irradiating dose and storage temperatures (p<0.05). Microbial counts for non-irradiated shrimp samples were higher than the respective irradiated samples at both storage temperatures (p<0.05). The results revealed that irradiation at high dose (5 kGy) might enhance lipid oxidation, although the growth of microorganisms and protein oxidation was inhibited.     

Effects of gamma irradiation on chemical composition and ruminal protein degradation of canola meal

Gamma irradiation of canola meal (at doses of 25, 50 and 75 kGy) could alter its ruminal protein degradation characteristics by cross-linking of the polypeptide chains. This processing resulted in decrease (linear effect, P<0.001) of ruminal protein degradation and increase (linear effect, P<0.001) of intestinal protein digestibility. The results showed that gamma irradiation at doses higher than 25 kGy can be used as a cross-linking agent to improve protein properties of supplements in ruminant nutrition.