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Sample records for guehomyces pullulans 17-1

  1. Laccases from Aureobasidium pullulans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Laccases are polyphenol oxidases (EC 1.10.3.2) that have numerous industrial and bioremediation applications. Laccases are well known as lignin-degrading enzymes, but these enzymes can play numerous other roles in fungi. In this study, 41 strains of the fungus Aureobasidium pullulans were examined f...

  2. Intraspecific diversity of Aureobasidium pullulans strains from different marine environments

    NASA Astrophysics Data System (ADS)

    Liu, Jia; Liu, Zhiqiang; Chi, Zhenming; Zhang, Liang; Zhang, Dechao

    2009-09-01

    Totally more than 500 yeast strains were isolated from seawater, sea sediments, mud of sea salterns, marine fish guts and marine algae. The results of routine and molecular biology identification methods show that nine strains among these marine yeasts belong to Aureobasidium pullulans, although the morphologies of their colonies are very different. The marine yeasts isolated from different marine environments indicate that A. pullulans is widely distributed in different environmental conditions. These Aureobasidium pullulans strains include A. pullulans 4#2, A. pullulans N13d, A. pullulans HN3-11, A. pullulans HN2-3, A. pullulans JHSc, A. pullulans HN4.7, A. pullulans HN5.3, A. pullulans HN6.2 and A. pullulans W13a. A. pullulans 4#2 could produce cellulase and single cell protein. A. pullulans N13d could produce protease, lipase, amylase and cellulase. Both A. pullulans HN3-11 and A. pullulans HN2-3 were able to produce protease, lipase and cellulase. A. pullulans JHSc could secrete cellulase and killer toxin. Both A. pullulans HN4.7 and A. pullulans HN5.3 could yield lipase and cellulase. A. pullulans W13a was able to secrete extracellular amylase and cellulase while A. pullulans HN4.7 and A. pullulans N13d could produce siderophores. This means that different A. pullulans strains from different marine environments have different physiological characteristics, which may be applied in many different biotechnological industries.

  3. 77 FR 8731 - Aureobasidium pullulans

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-02-15

    ...-5805. II. Background and Statutory Findings In the Federal Register of March 10, 2010 (75 FR 11171... results of a study of the influence of temperature on the growth of the two Aureobasidium pullulans strains, which showed that growth of both strains does not occur at or above 35 C. Since human...

  4. Statistical modeling of pullulan production and its application in pullulan acetate nanoparticles synthesis.

    PubMed

    Sugumaran, K R; Ponnusami, V

    2015-11-01

    This work focused on the optimization of exo-polysaccharide, pullulan production by exploiting cassava bagasse, an agricultural solid waste residue by solid state fermentation and its application in the preparation of pullulan acetate nanoparticles. Statistical approach was investigated to maximize the pullulan production using C/N ratio, initial pH, NaCl and ZnSO4·5H2O. The optimum conditions for maximum yield of pullulan (39.42±0.62mg/gds) were found to be: C/N=25.94, initial pH=5.5 and NaCl=0.55g/L. Using the optimized medium variables, the production of pullulan was investigated in lab scale solid state fermentation. The pullulan produced was characterized by thermo gravimetric and XRD analysis. Also pullulan acetate nanoparticles were synthesized from chemical modification of pullulan and the average particle size of nanoparticles was examined by zeta particle sizer.

  5. Color Variability, Pullulan Production, and Pullulan Degrading Enzyme Activities in Tropical Isolates of Aureobasidium Pullulans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Forty-six isolates from fifteen provinces in Thailand of the polymorphic fungus Aureobasidium pullulans were cultured. They were isolated from leaves, painted wall and wood surfaces. Each isolate was cultured in production medium (PM) containing sucrose and peptone as carbon and nitrogen sources, ...

  6. a-Amylase activity during pullulan production and a-Amylase gene analyses of Aureobasidium pullulans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The fungus Aureobasidium pullulans is the source of commercially produced pullulan, a high molecular weight polysaccharide that is used in the manufacture of edible films. It has been proposed that alpha-amylase negatively affects the molecular weight of pullulan in late cultures. Based on a recen...

  7. The mechanism of improved pullulan production by nitrogen limitation in batch culture of Aureobasidium pullulans.

    PubMed

    Wang, Dahui; Chen, Feifei; Wei, Gongyuan; Jiang, Min; Dong, Mingsheng

    2015-08-20

    Batch culture of Aureobasidium pullulans CCTCC M 2012259 for pullulan production at different concentrations of ammonium sulfate and yeast extract was investigated. Increased pullulan production was obtained under nitrogen-limiting conditions, as compared to that without nitrogen limitation. The mechanism of nitrogen limitation favoring to pullulan overproduction was revealed by determining the activity as well as gene expression of key enzymes, and energy supply for pullulan biosynthesis. Results indicated that nitrogen limitation increased the activities of α-phosphoglucose mutase and glucosyltransferase, up-regulated the transcriptional levels of pgm1 and fks genes, and supplied more ATP intracellularly, which were propitious to further pullulan biosynthesis. The economic analysis of batch pullulan production indicated that nitrogen limitation could reduce more than one third of the cost of raw materials when glucose was supplemented to a total concentration of 70 g/L. This study also helps to understand the mechanism of other polysaccharide overproduction by nitrogen limitation.

  8. Copper sulfate improves pullulan production by bioconversion using whole cells of Aureobasidium pullulans as the catalyst.

    PubMed

    Wang, Dahui; Ju, Xiaomin; Zhang, Gaochuan; Wang, Donghua; Wei, Gongyuan

    2016-10-01

    The effects of mineral salts on pullulan production by bioconversion using whole cells of Aureobasidium pullulans CCTCC M 2012259 as the catalyst were investigated. Copper sulfate (CuSO4) improved pullulan production by 36.2% and 42.3% when added at the optimum concentration of 0.2mg/L to the bioconversion broth or seed medium, respectively, as compared with controls without CuSO4 addition. Pullulan production was further enhanced when CuSO4 was added to both seed medium and bioconversion broth simultaneously. In order to probe the mechanism of CuSO4 improvement, cell viability, membrane integrity, intracellular adenosine triphosphate (ATP) levels and the activities of key enzymes involved in pullulan biosynthesis were determined. As a result, CuSO4 increased the activities of key biosynthetic enzymes, maintained intracellular ATP at a higher level, and accelerated the rate of pullulan secretion, all of which contributed to improved pullulan production by bioconversion.

  9. HLC/pullulan and pullulan hydrogels: their microstructure, engineering process and biocompatibility.

    PubMed

    Li, Xian; Xue, Wenjiao; Liu, Yannan; Li, Weina; Fan, Daidi; Zhu, Chenhui; Wang, Yaoyu

    2016-01-01

    New locally injectable biomaterials that are suitable for use as soft tissue fillers are needed to address a significant unmet medical need. In this study, we used pullulan and human-like collagen (HLC) based hydrogels with various molecular weights (MWs) in combination therapy against tissue defects. Briefly, pullulan was crosslinked with NaIO4 to form a pullulan hydrogel and then may coupled with HLC using the reaction between the -NH2 end-group of HLC and the -CHO group present on the aldehyde pullulan to form the HLC/pullulan hydrogel, wherein the NaIO4 acted as the crosslinking and oxidizing agent. The good miscibility of pullulan and HLC in the hydrogels was confirmed via Fourier transform infrared spectroscopy, scanning electron microscopy, compression testing, enzyme degradation testing, cell adhesions, live/dead staining and subcutaneous filling assays. Here, pullulan hydrogels with various MWs were fabricated and physicochemically characterized. Limitations of the pullulan hydrogels included inflammation, poor mechanical strength, and degradation. By contrast, the properties of the HLC/pullulan hydrogels strongly enhanced. The efficacy of these hydrogels was evaluated both in vitro and in vivo. Our results indicate that HLC/pullulan hydrogels may have therapeutic value as efficient soft tissue fillers, with reduced inflammation, improved cell adhesion and delayed hydrogel degradation.

  10. HLC/pullulan and pullulan hydrogels: their microstructure, engineering process and biocompatibility.

    PubMed

    Li, Xian; Xue, Wenjiao; Liu, Yannan; Li, Weina; Fan, Daidi; Zhu, Chenhui; Wang, Yaoyu

    2016-01-01

    New locally injectable biomaterials that are suitable for use as soft tissue fillers are needed to address a significant unmet medical need. In this study, we used pullulan and human-like collagen (HLC) based hydrogels with various molecular weights (MWs) in combination therapy against tissue defects. Briefly, pullulan was crosslinked with NaIO4 to form a pullulan hydrogel and then may coupled with HLC using the reaction between the -NH2 end-group of HLC and the -CHO group present on the aldehyde pullulan to form the HLC/pullulan hydrogel, wherein the NaIO4 acted as the crosslinking and oxidizing agent. The good miscibility of pullulan and HLC in the hydrogels was confirmed via Fourier transform infrared spectroscopy, scanning electron microscopy, compression testing, enzyme degradation testing, cell adhesions, live/dead staining and subcutaneous filling assays. Here, pullulan hydrogels with various MWs were fabricated and physicochemically characterized. Limitations of the pullulan hydrogels included inflammation, poor mechanical strength, and degradation. By contrast, the properties of the HLC/pullulan hydrogels strongly enhanced. The efficacy of these hydrogels was evaluated both in vitro and in vivo. Our results indicate that HLC/pullulan hydrogels may have therapeutic value as efficient soft tissue fillers, with reduced inflammation, improved cell adhesion and delayed hydrogel degradation. PMID:26478402

  11. Central metabolic pathways of Aureobasidium pullulans CGMCC1234 for pullulan production.

    PubMed

    Sheng, Long; Liu, Chang; Tong, Qunyi; Ma, Meihu

    2015-12-10

    With the purpose of understanding the metabolic network of Aureobasidium pullulans, the central metabolic pathways were confirmed by the activities of the key enzymes involved in different pathways. The effect of different iodoacetic acid concentrations on pullulan fermentation was also investigated in this paper. The activities of phosphofructokinases and glucose-6-phosphate dehydrogenase existed in A. pullulans CGMCC1234, whereas 2-keto-3-deoxy-6-phosphogluconate aldolase activity was not detected. We proposed that the central metabolic pathways of A. pullulans CGMCC1234 included EMP and PPP, but no ED. Pullulan production declined fast as the iodoacetic acid increased, while cell growth offered upgrade firstly than descending latter tendency. Compared to the control group, the ratio of ATP/ADP of 0.60 mM iodoacetic acid group was lower at different stages of pullulan fermentation. The findings revealed that low concentration of iodoacetic acid might impel carbon flux flow toward the PPP, but reduce the flux of the EMP.

  12. Why sucrose is the most suitable substrate for pullulan fermentation by Aureobasidium pullulans CGMCC1234?

    PubMed

    Sheng, Long; Tong, Qunyi; Ma, Meihu

    2016-10-01

    This paper studies the metabolic pathway of sucrose in pullulan fermentation by Aureobasidium pullulans. Because of its high pullulan production, sucrose proved to be the best carbon source for pullulan synthesis by A. pullulans CGMCC1234 (36.3g/L). Compared to other carbon sources, A. pullulans cells reached the stationary phase more quickly in the presence of sucrose. The specific sugar types and concentrations occurring during pullulan fermentation were detected using High Performance Liquid Chromatography (HPLC). HPLC results revealed that sucrose did not simply break down into glucose and fructose in the medium employed. Kestose (22.69g/L) also accumulated during early stages of fermentation (24h), which reduced the osmotic pressure of the extracellular fluid and diminished the inhibition of cell growth and pullulan production. β-Fructofuranosidase activity strongly depended on the carbon source. Sucrose was the best inducer of β-fructofuranosidase production. However, β-fructofuranosidase production did not directly and/or proportionally correlate with the growth of A. pullulans cells. PMID:27542744

  13. Production of high molecular weight pullulan by Aureobasidium pullulans HP-2001 with soybean pomace as a nitrogen source.

    PubMed

    Seo, Hyung-Pil; Son, Chang-Woo; Chung, Chung-Han; Jung, Dae-Il; Kim, Sung-Koo; Gross, Richard A; Kaplan, David L; Lee, Jin-Woo

    2004-12-01

    The production of pullulan by Aureobasidium pullulans HP-2001 was enhanced by yeast extract as a nitrogen source as well as soybean pomace. The highest production of pullulan by A. pullulans HP-2001 with yeast extract was 5.5 g/l whereas that of pullulan with soybean pomace was 7.5 g/l. The gas chromatogram of pullulan produced by A. pullulans HP-2001 with soybean pomace as a nitrogen source showed that the major and minor components were glucose and mannose. The FTIR spectra of pullulans produced with yeast extract, a mixture of yeast extract and soybean pomace, and soybean pomace alone exhibited similar features. The increase in content of reducing sugars after pullulanase treatment of pullulans produced with different nitrogen sources indicated that all the pullulans had alpha-(1,6) glucosidic linkages of alpha-(1,4) linked maltotriose units. The average molecular weights of pullulans produced with various concentrations of yeast extract and soybean pomace ranged from 0.17 to 1.32x10(6) and from 1.32 to 5.66x10(6), respectively. All pullulans produced by A. pullulans HP-2001 in this study had the same basic structures, but their ratios of monomeric components were a little different, which might result in the production of pullulans with different molecular weights.

  14. Aureobasidium pullulans xylanase, gene and signal sequence

    DOEpatents

    Xin-Liang, Li; Ljungdahl, Lars G.

    1997-01-01

    A xylanase from Aureobasidium pullulans having a high specific activity is provided as well as a signal protein for controlling excretion into cell culture medium of proteins to which it is attached. DNA encoding these proteins is also provided.

  15. Multilocus Phylogenetic Analyses, Pullulan Production and Xylanase Activity of Tropical Isolates of Aureobasidium pullulans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aureobasidium pullulans is the source of the commercial polysaccharide, pullulan, and the enzyme, xylanase (EC 3.2.1.8). Isolates are typically off-white to black on solid media, while some tropical isolates have been described as "color variants" with bright pigments of red, yellow, or purple. In...

  16. Production of pullulan by Aureobasidium pullulans from Asian palm kernel: a novel substrate.

    PubMed

    Sugumaran, K R; Gowthami, E; Swathi, B; Elakkiya, S; Srivastava, S N; Ravikumar, R; Gowdhaman, D; Ponnusami, V

    2013-01-30

    Production of a commercially important biodegradable polymer, pullulan, by Aureobasidium pullulans from four agricultural wastes namely wheat bran, rice bran, coconut kernel and palm kernel was evaluated in solid state fermentation. Under the experimental conditions, palm kernel resulted in highest concentration of pullulan (16 g/L) among the four solid substrates. Optimum initial pH and moisture content for pullulan production were found out to be 6.5 and 50% respectively. 18.43 g/L of pullulan was produced from Asian palm kernel with initial pH 6.5 after 7 days of fermentation and yeast like morphology was predominant under this condition. Among different nitrogen sources tried in this study, yeast extract was found to the best. The pullulan produced from palm kernel was characterized by FTIR and (1)H NMR. The results were matching with that of commercial pullulan. Thus, Asian palm kernel appears to be an attractive low cost carbon source for the production of pullulan. PMID:23218356

  17. Pullulan Production by Aureobasidium pullulans ATCC 201253 Cells Adsorbed onto Cellulose Anion and Cation Exchangers

    PubMed Central

    West, Thomas P.

    2012-01-01

    The anion exchanger phosphocellulose and the cation exchanger triethylaminoethyl cellulose were used to immobilize cells of the fungus Aureobasidium pullulans ATCC 201253 and the adsorbed cells were subsequently investigated for their ability to produce the polysaccharide pullulan using batch fermentation. The cells adsorbed on the triethylaminoethyl cellulose at pH 7.5 produced higher pullulan levels than those cells immobilized on phosphocellulose at pH 4.0 for 2 cycles of 168 h at 30 °C. Relative to the initial cycle of 168 h, pullulan production by the cells immobilized on the triethylaminoethyl cellulose decreased slightly after 168 h of the second production cycle while pullulan production by the phosphocellulose-immobilized cells remained about the same after 168 h of the second production cycle. PMID:23762749

  18. Relationship Between a-amylase Activity and Pullulan Profiles, and a-amylase Gene Analyses of the Fungus Aureobasidium Pullulans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Tropical isolates of Aureobasidium pullulans isolated from various habitats in Thailand were classified based on multilocus phylogenetic analyses using concordance analysis of DNA sequences. This fungus is the major source of commercially produced pullulan, a high molecular weight polysaccharide th...

  19. Copper sulfate improves pullulan production by bioconversion using whole cells of Aureobasidium pullulans as the catalyst.

    PubMed

    Wang, Dahui; Ju, Xiaomin; Zhang, Gaochuan; Wang, Donghua; Wei, Gongyuan

    2016-10-01

    The effects of mineral salts on pullulan production by bioconversion using whole cells of Aureobasidium pullulans CCTCC M 2012259 as the catalyst were investigated. Copper sulfate (CuSO4) improved pullulan production by 36.2% and 42.3% when added at the optimum concentration of 0.2mg/L to the bioconversion broth or seed medium, respectively, as compared with controls without CuSO4 addition. Pullulan production was further enhanced when CuSO4 was added to both seed medium and bioconversion broth simultaneously. In order to probe the mechanism of CuSO4 improvement, cell viability, membrane integrity, intracellular adenosine triphosphate (ATP) levels and the activities of key enzymes involved in pullulan biosynthesis were determined. As a result, CuSO4 increased the activities of key biosynthetic enzymes, maintained intracellular ATP at a higher level, and accelerated the rate of pullulan secretion, all of which contributed to improved pullulan production by bioconversion. PMID:27312631

  20. Production of pullulan from raw potato starch hydrolysates by a new strain of Auerobasidium pullulans.

    PubMed

    Wu, Shengjun; Lu, Mingsheng; Chen, Jing; Fang, Yaowei; Wu, Leilei; Xu, Yan; Wang, Shujun

    2016-01-01

    In the present study, hydrolysis of potato starch with marine cold-adapted α-amylase and pullulan production from the hydrolysates by a new strain of Auerobasidium pullulans isolated from sea mud were conducted. The hydrolysis conditions were optimized as follows: reaction time 2h, pH 6.5, temperature 20°C, and α-amylase amount 12 U/g. Under these optimum hydrolysis conditions, the DE value of the potato starch hydrolysates reached to 49.56. The potato starch hydrolysates consist of glucose, maltose, isomaltose, maltotriose, and trace of other maltooligosaccharides with degree of polymerization ranged 4-7. The maximum production of pullulan at 96 h from the hydrolysate of potato starch was 36.17 g/L, which was higher than those obtained from glucose (22.07 g/L, p<0.05) and sucrose (31.42 g/L, p<0.05). Analysis of the high performance liquid chromatography of the hydrolysates of the pullulan product with pullulanase indicated that the main composition is maltotriose, thus confirming the pullulan structure of this pullulan product.

  1. Central metabolic pathways of Aureobasidium pullulans CGMCC1234 for pullulan production.

    PubMed

    Sheng, Long; Liu, Chang; Tong, Qunyi; Ma, Meihu

    2015-12-10

    With the purpose of understanding the metabolic network of Aureobasidium pullulans, the central metabolic pathways were confirmed by the activities of the key enzymes involved in different pathways. The effect of different iodoacetic acid concentrations on pullulan fermentation was also investigated in this paper. The activities of phosphofructokinases and glucose-6-phosphate dehydrogenase existed in A. pullulans CGMCC1234, whereas 2-keto-3-deoxy-6-phosphogluconate aldolase activity was not detected. We proposed that the central metabolic pathways of A. pullulans CGMCC1234 included EMP and PPP, but no ED. Pullulan production declined fast as the iodoacetic acid increased, while cell growth offered upgrade firstly than descending latter tendency. Compared to the control group, the ratio of ATP/ADP of 0.60 mM iodoacetic acid group was lower at different stages of pullulan fermentation. The findings revealed that low concentration of iodoacetic acid might impel carbon flux flow toward the PPP, but reduce the flux of the EMP. PMID:26428132

  2. Aureobasidium pullulans xylanase, gene and signal sequence

    DOEpatents

    Li Xinliang; Ljungdahl, L.G.

    1997-01-07

    A xylanase from Aureobasidium pullulans having a high specific activity is provided, as well as a signal protein for controlling excretion into cell culture medium of proteins to which it is attached. DNA encoding these proteins is also provided. 4 figs.

  3. Peat hydrolysate medium optimization for pullulan production.

    PubMed

    Boa, J M; Leduy, A

    1984-07-01

    Peat hydrolysate, a diluted acid-autoclaved extract of peat, was used as a substrate for the production of the extracellular polysaccharide pullulan by three strains of Aureobasidium pullulans, 140B, 142, and 2552. It was found that the addition of (NH(4))(2)SO(4) and K(2)HPO(4) as sources of nitrogen and phosphate, respectively, is not necessary for the polysaccharide production. The economically optimized culture medium for large-scale production of pullulan contains peat hydrolysate, 0.05% NaCl, 0.02% MgSO(4), and 0.01% antifoam FG-10. The initial pH of peat hydrolysate medium is adjusted to its optimum value of 6.0 with Ca(OH)(2). The total ingredient cost for the production of each kilogram of pullulan with optimized medium is only 1/10 of that with the nonoptimized medium. In this study, a zero cost for peat hydrolysate was assumed, since it is an effluent of the peat and peat processing industries.

  4. Heavy Oils Produced by Aureobasidium pullulans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    From a survey of more than 50 diverse strains of Aureobasidium pullulans, 21 strains were observed to produce extracellular heavy oils. These strains represented at least 6 phylogenetic clades, although more than half fell into clades 9 and 11. Oil colors ranged from bright yellow to malachite. M...

  5. Dichromated pullulan as a novel photosensitive holographic material.

    PubMed

    Panteli, D; Savi, S; Jakovljevic, D

    1998-05-15

    We found that pullulan [an a -glucan with (1?4) and (1?6) linkages] doped with chromium ions posseses promising holographic properties. It is environmentally stable and is capable of registering high spatial frequencies (as many as 1500 lines/mm) and producing surface relief. At 1000 lines/mm pullulan's diffraction efficiency was 4%. Chemical treatment of pullulan with a mixture of water and isopropyl alcohol is simple. However, as expected from analogy with dichromated gelatin, pullulan is sensitive in only the blue-green region of the spectrum. PMID:18087349

  6. Silk fibroin/pullulan blend films: Preparation and characterization

    NASA Astrophysics Data System (ADS)

    Shivananda, C. S.; Rao, B. Lakshmeesha; Madhukumar, R.; Sarojini, B. K.; Somashekhar, R.; Asha, S.; Sangappa, Y.

    2016-05-01

    In this work silk fibroin/pullulan blend films have been prepared by solution casting method. The blend films were examined for structural, and thermal properties using X-ray diffraction (XRD), thermogravimatric (TGA) and differential scanning calorimetry (DSC) analysis. The XRD results indicate that with the introduction of pullulan, the interaction between SF and pullulan in the blend films induced the conformation transition of SF films and amorphous phase increases with increasing pullulan ratio. The thermal properties of the blend films were improved significantly in the blend films.

  7. Pullulan-hyperproducing color variant strain of Aureobasidium pullulans FB-1 newly isolated from phylloplane of Ficus sp.

    PubMed

    Singh, R S; Saini, G K

    2008-06-01

    The studies were carried out for the isolation of efficient pullulan producing strains of Aureobasidium pullulans. Five strains were isolated from phylloplane of different plants. Amongst these, three were producing black pigment melanin, while the remaining two produced pink pigment. These two color variant isolates of A. pullulans were designated as FB-1 and FG-1, and obtained from phylloplane of Ficus benjamina and Ficus glometa, respectively. The parameters employed for the identification of the isolates included morphology, nutritional assimilation patterns and exopolysaccharide (EPS) production. Isolates were compared with standard cultures for EPS production. A. pullulans FB-1 was the best producer of pullulan giving up to 1.9, 1.4 and 1.7 times more pullulan than the control of A. pullulans NCIM 976, NCIM 1048 and NCIM 1049, respectively. The IR spectra of the isolates and standard strains revealed that the polysaccharide was pullulan, but not aubasidan. The study also supported the fact that A. pullulans is a ubiquitous organism and phylloplane being the important niche of the organism. PMID:17869506

  8. Pullulan from peat hydrolyzate fermentation kinetics.

    PubMed

    Boa, J M; Leduy, A

    1987-09-01

    The Luedeking-Piret equation was used to fit the kinetic data of pullulan fermentations from peat hydrolyzate substrate. In batch mode, the kinetic parameters m, n, alpha, and beta varied as a function of fermentation conditions: aeration rate, agitation speed, and temperature. In constant-feed fed-batch mode, the parameters Varied according to the feed rates. In peat hydrolyzate medium, the polysaccharide synthesis was strongly growth associated in batch and continuous fermentations but entirely growth associated in fedbatch fermentations. The fed-batch mode of fermentation with an appropriate feed rate is more advantageous with respect to batch and continuous fermentations. Therefore, if the fermentation is started batchwise and then followed by fed-batch mode at a constant feed rate, the overall polysaccharide productivity (g pullulan/L h) is significantly higher than those obtained with batch or continuous fermentations using the same total medium volume.

  9. Multilocus phylogenetic analyses, pullulan production and xylanase activity of tropical isolates of Aureobasidium pullulans.

    PubMed

    Manitchotpisit, Pennapa; Leathers, Timothy D; Peterson, Stephen W; Kurtzman, Cletus P; Li, Xin-Liang; Eveleigh, Douglas E; Lotrakul, Pongtharin; Prasongsuk, Sehanat; Dunlap, Christopher A; Vermillion, Karl E; Punnapayak, Hunsa

    2009-10-01

    Aureobasidium pullulans is the source of the commercially valuable polysaccharide pullulan and the enzyme xylanase. Isolates are typically off-white to pale pink or black on solid media, while some tropical isolates have been described as 'color variants' with bright pigments of red, yellow or purple. We sequenced 5 loci (internal transcribed spacer, intergenic spacer 1, translation elongation factor-1 alpha, beta tubulin, and RNA polymerase II) from 45 new isolates from Thailand. Based on the phylogenetic analyses, isolates were classified into 12 clades. Each clade showed different colors on different culture media including two clades with 'color variants' and some clades exhibited high levels of pullulan production or xylanase activity. Colony characteristics do not correlate perfectly with DNA sequence phylogeny or the physiological characters, but DNA sequence differences rapidly identify isolates with genetic novelty. PMID:19619651

  10. An extremely thermostable amylopullulanase from Staphylothermus marinus displays both pullulan- and cyclodextrin-degrading activities.

    PubMed

    Li, Xiaolei; Li, Dan; Park, Kwan-Hwa

    2013-06-01

    A gene encoding an amylopullulanase of the glycosyl hydrolase (GH) family 57 from Staphylothermus marinus (SMApu) was heterologously expressed in Escherichia coli. SMApu consisted of 639 amino acids with a molecular mass of 75.3 kDa. It only showed maximal amino acid identity of 17.1 % with that of Pyrococcus furiosus amylopullulanase in all identified amylases. Not like previously reported amylopullulanases, SMApu has no signal peptide but contains a continuous GH57N_Apu domain. It had the highest catalytic efficiency toward pullulan (k cat/K m , 342.34 s(-1) mL mg(-1)) and was extremely thermostable with maximal pullulan-degrading activity (42.1 U/mg) at 105 °C and pH 5.0 and a half-life of 50 min at 100 °C. Its activity increased to 116 % in the presence of 5 mM CaCl2. SMApu could also degrade cyclodextrins, which are resistant to the other amylopullulanases. The initial hydrolytic products from pullulan, γ-CD, and 6-O-maltooligosyl-β-CD were [6)-α-D-Glcp-(1 → 4)-α-D-Glcp-(1 → 4)-α-D-Glcp-(1→]n, maltooctaose, and single maltooligosaccharide plus β-CD, respectively. The final hydrolytic products from above-mentioned substrates were maltose and glucose. These results confirm that SMApu is a novel amylopullulanase of the family GH57 possessing the cyclodextrin-degrading activity of cyclomaltodextrinase. PMID:23001056

  11. An extremely thermostable amylopullulanase from Staphylothermus marinus displays both pullulan- and cyclodextrin-degrading activities.

    PubMed

    Li, Xiaolei; Li, Dan; Park, Kwan-Hwa

    2013-06-01

    A gene encoding an amylopullulanase of the glycosyl hydrolase (GH) family 57 from Staphylothermus marinus (SMApu) was heterologously expressed in Escherichia coli. SMApu consisted of 639 amino acids with a molecular mass of 75.3 kDa. It only showed maximal amino acid identity of 17.1 % with that of Pyrococcus furiosus amylopullulanase in all identified amylases. Not like previously reported amylopullulanases, SMApu has no signal peptide but contains a continuous GH57N_Apu domain. It had the highest catalytic efficiency toward pullulan (k cat/K m , 342.34 s(-1) mL mg(-1)) and was extremely thermostable with maximal pullulan-degrading activity (42.1 U/mg) at 105 °C and pH 5.0 and a half-life of 50 min at 100 °C. Its activity increased to 116 % in the presence of 5 mM CaCl2. SMApu could also degrade cyclodextrins, which are resistant to the other amylopullulanases. The initial hydrolytic products from pullulan, γ-CD, and 6-O-maltooligosyl-β-CD were [6)-α-D-Glcp-(1 → 4)-α-D-Glcp-(1 → 4)-α-D-Glcp-(1→]n, maltooctaose, and single maltooligosaccharide plus β-CD, respectively. The final hydrolytic products from above-mentioned substrates were maltose and glucose. These results confirm that SMApu is a novel amylopullulanase of the family GH57 possessing the cyclodextrin-degrading activity of cyclomaltodextrinase.

  12. 46 CFR 95.17-1 - Application.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 4 2010-10-01 2010-10-01 false Application. 95.17-1 Section 95.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CARGO AND MISCELLANEOUS VESSELS FIRE PROTECTION EQUIPMENT Foam Extinguishing Systems, Details § 95.17-1 Application. (a) Where a foam extinguishing...

  13. 46 CFR 95.17-1 - Application.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 4 2013-10-01 2013-10-01 false Application. 95.17-1 Section 95.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CARGO AND MISCELLANEOUS VESSELS FIRE PROTECTION EQUIPMENT Foam Extinguishing Systems, Details § 95.17-1 Application. (a) Where a foam extinguishing...

  14. 46 CFR 76.17-1 - Application.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 3 2012-10-01 2012-10-01 false Application. 76.17-1 Section 76.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) PASSENGER VESSELS FIRE PROTECTION EQUIPMENT Foam Extinguishing Systems, Details § 76.17-1 Application. (a) Where a foam extinguishing system is installed,...

  15. 46 CFR 76.17-1 - Application.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 3 2011-10-01 2011-10-01 false Application. 76.17-1 Section 76.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) PASSENGER VESSELS FIRE PROTECTION EQUIPMENT Foam Extinguishing Systems, Details § 76.17-1 Application. (a) Where a foam extinguishing system is installed,...

  16. 46 CFR 76.17-1 - Application.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 3 2010-10-01 2010-10-01 false Application. 76.17-1 Section 76.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) PASSENGER VESSELS FIRE PROTECTION EQUIPMENT Foam Extinguishing Systems, Details § 76.17-1 Application. (a) Where a foam extinguishing system is installed,...

  17. 46 CFR 95.17-1 - Application.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 4 2014-10-01 2014-10-01 false Application. 95.17-1 Section 95.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CARGO AND MISCELLANEOUS VESSELS FIRE PROTECTION EQUIPMENT Foam Extinguishing Systems, Details § 95.17-1 Application. (a) Where a foam extinguishing...

  18. 46 CFR 76.17-1 - Application.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 3 2013-10-01 2013-10-01 false Application. 76.17-1 Section 76.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) PASSENGER VESSELS FIRE PROTECTION EQUIPMENT Foam Extinguishing Systems, Details § 76.17-1 Application. (a) Where a foam extinguishing system is installed,...

  19. 46 CFR 95.17-1 - Application.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 4 2012-10-01 2012-10-01 false Application. 95.17-1 Section 95.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CARGO AND MISCELLANEOUS VESSELS FIRE PROTECTION EQUIPMENT Foam Extinguishing Systems, Details § 95.17-1 Application. (a) Where a foam extinguishing...

  20. 46 CFR 76.17-1 - Application.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 3 2014-10-01 2014-10-01 false Application. 76.17-1 Section 76.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) PASSENGER VESSELS FIRE PROTECTION EQUIPMENT Foam Extinguishing Systems, Details § 76.17-1 Application. (a) Where a foam extinguishing system is installed,...

  1. 46 CFR 95.17-1 - Application.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 4 2011-10-01 2011-10-01 false Application. 95.17-1 Section 95.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CARGO AND MISCELLANEOUS VESSELS FIRE PROTECTION EQUIPMENT Foam Extinguishing Systems, Details § 95.17-1 Application. (a) Where a foam extinguishing...

  2. 43 CFR 17.1 - Purpose.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 43 Public Lands: Interior 1 2012-10-01 2011-10-01 true Purpose. 17.1 Section 17.1 Public Lands... DEPARTMENT OF THE INTERIOR Nondiscrimination on the Basis of Race, Color, or National Origin § 17.1 Purpose... to the end that no person in the United States shall, on the grounds of race, color, or...

  3. Multilocus phylogenetic analyses and phenotypic characterization of tropical isolates of Aureobasidium pullulans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aureobasidium pullulans is the source of the commercial polysaccharide, pullulan, and the enzyme, xylanase. The purpose of this study was to classify Aureobasidium isolates using multilocus phylogenetic analysis and determine specific characteristics of each clade....

  4. Bioconversion of industrial solid waste--cassava bagasse for pullulan production in solid state fermentation.

    PubMed

    Sugumaran, K R; Jothi, P; Ponnusami, V

    2014-01-01

    The purpose of the work was to produce commercially important pullulan using industrial solid waste namely cassava bagasse in solid state fermentation and minimize the solid waste disposal problem. First, influence of initial pH on cell morphology and pullulan yield was studied. Effect of various factors like fermentation time, moisture ratio, nitrogen sources and particle size on pullulan yield was investigated. Various supplementary carbon sources (3%, w/w) namely glucose, sucrose, fructose, maltose, mannose and xylose with cassava bagasse was also studied to improve the pullulan yield. After screening the suitable supplement, effect of supplement concentration on pullulan production was investigated. The pullulan from cassava bagasse was characterized by FTIR, (1)H-NMR and (13)C-NMR. Molecular weight of pullulan from cassava bagasse was determined by gel permeation chromatography. Thus, cassava bagasse emerged to be a cheap and novel substrate for pullulan production.

  5. Liamocins from Aureobasidium pullulans: New and highly selective anti-streptococcal agents

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The ubiquitous, black yeast Aureobasidium pullulans, which is used commercially to produce pullulan polysaccharide, also produces several polyol-lipids that we have named liamocins. A survey of >50 strains of A. pullulans identified 21 diverse strains that produce liamocins. Typically the liamocins ...

  6. Recent insights on applications of pullulan in tissue engineering.

    PubMed

    Singh, Ram Sarup; Kaur, Navpreet; Rana, Vikas; Kennedy, John F

    2016-11-20

    Tissue engineering is a recently emerging line of act which assists the regeneration of damaged tissues, unable to self-repair themselves and in turn, enhances the natural healing potential of patients. The repair of injured tissue can be induced with the help of some artificially created polymer scaffolds for successful tissue regeneration. The pullulan composite scaffolds can be used to enhance the proliferation and differentiation of cells for tissue regeneration. The unique pattern of pullulan with α-(1→4) and α-(1→6) linkages along with the presence of nine hydroxyl groups on its surface, endows the polymer with distinctive physical features required for tissue engineering. Pullulan can be used for vascular engineering, bone repair and skin tissue engineering. Pullulan composite scaffolds can also be used for treatment of injured femoral condyle bone, skull bone and full thickness skin wound of murine models, transversal mandibular and tibial osteotomy in goat, etc. This review article highlights the latest developments on applications of pullulan and its derivatives in tissue engineering. PMID:27561517

  7. Pullulan fermentation using a prototype rotational reciprocating plate impeller.

    PubMed

    Lin, Yun; Thibault, Jules

    2013-05-01

    A rotational reciprocating plate impeller prototype, designed to improve the mixing homogeneity of viscous non-Newtonian fermentation broth, has been tested in pullulan fermentations. With this new impeller, the operating levels of several factors were investigated to improve pullulan production with Aureobasidium pullulans ATCC 42023 in a 22-L bioreactor using experimental designs. Because both high molecular weight (MW) and high concentration of pullulan were desired; the exopolysaccharide (EPS) concentration and the broth viscosity were used as optimization objective functions to be maximized. A 6-run uniform design was used to investigate five factors. Under the best operating conditions among the six runs, 29.0 g L(-1) EPS was produced at 102 h. This condition was used as the starting point for further investigation on the two statistically significant factors, the pH and the agitation speed. An 8-run 3-level custom design that investigates up to second-order effects was used in the second stage. An optimal zone of operating conditions for large quantity of high MW pullulan production was identified. A concentration of 23.3 g L(-1) EPS was produced at 78 h. This is equivalent to an EPS productivity of 0.30 g L(-1) h(-1). The corresponding apparent viscosity of the broth was 0.38 Pa s at the shear rate of 10 s(-1). PMID:22940838

  8. Pullulan microcarriers for bone tissue regeneration.

    PubMed

    Aydogdu, Hazal; Keskin, Dilek; Baran, Erkan Turker; Tezcaner, Aysen

    2016-06-01

    Microcarrier systems offer a convenient way to repair bone defects as injectable cell carriers that can be applied with small incisions owing to their small size and spherical shape. In this study, pullulan (PULL) microspheres were fabricated and characterized as cell carriers for bone tissue engineering applications. PULL was cross-linked by trisodium trimetaphosphate (STMP) to enhance the stability of the microspheres. Improved cytocompatibility was achieved by silk fibroin (SF) coating and biomimetic mineralization on the surface by incubating in simulated body fluid (SBF). X-ray diffraction (XRD), scanning electron microscopy (SEM) and fluorescent microscopy analysis confirmed biomimetic mineralization and SF coating on microspheres. The degradation analysis revealed that PULL microspheres had a slow degradation rate with 8% degradation in two weeks period indicating that the microspheres would support the formation of new bone tissue. Furthermore, the mechanical tests showed that the microspheres had a high mechanical stability that was significantly enhanced with the biomimetic mineralization. In vitro cell culture studies with SaOs-2 cells showed that cell viability was higher on SF and SBF coated microspheres on 7th day compared to PULL ones under dynamic conditions. Alkaline phosphatase activity was higher for SF coated microspheres in comparison to uncoated microspheres when dynamic culture condition was applied. The results suggest that both organic and inorganic surface modifications can be applied on PULL microspheres to prepare a biocompatible microcarrier system with suitable properties for bone tissue engineering. PMID:27040238

  9. 15 CFR 17.1 - Licensing rules.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 15 Commerce and Foreign Trade 1 2010-01-01 2010-01-01 false Licensing rules. 17.1 Section 17.1 Commerce and Foreign Trade Office of the Secretary of Commerce LICENSING OF GOVERNMENT-OWNED INVENTIONS IN THE CUSTODY OF THE DEPARTMENT OF COMMERCE Licensing of Rights in Domestic Patents and...

  10. 36 CFR 17.1 - Authority.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... Act of July 15, 1968, 82 Stat. 354, 16 U.S.C. 4601-22(a), authorizes the Secretary of the Interior... 36 Parks, Forests, and Public Property 1 2011-07-01 2011-07-01 false Authority. 17.1 Section 17.1 Parks, Forests, and Public Property NATIONAL PARK SERVICE, DEPARTMENT OF THE INTERIOR CONVEYANCE...

  11. 22 CFR 17.1 - General.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 22 Foreign Relations 1 2010-04-01 2010-04-01 false General. 17.1 Section 17.1 Foreign Relations DEPARTMENT OF STATE PERSONNEL OVERPAYMENTS FROM THE FOREIGN SERVICE RETIREMENT AND DISABILITY FUND UNDER THE FOREIGN SERVICE RETIREMENT AND DISABILITY SYSTEM (FSRDS) AND THE FOREIGN SERVICE PENSION SYSTEM...

  12. Production of novel antibacterial liamocins by strains of Aureobasidium pullulans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Certain strains of Aureobasidium pullulans produce liamocins, heavier-than-water “oils” that accumulate in liquid cultures. Liamocins are surface active, and inhibit mammalian cancer cell lines. Recently, we discovered that liamocins have antibacterial activity with specificity against Streptococcus...

  13. Isolation and Identification of Trehalase from Pullularia pullulans

    PubMed Central

    Merdinger, Emanuel; Lange, Charles F.; Booker, Ben F.

    1971-01-01

    Trehalase has been isolated from Pullularia pullulans. The enzyme, which is specific for trehalose, was purified approximately 800-fold. The optimal pH was found to be 4.0 and the Michaelis dissociation constant, Km, was determined to be 3.2 × 10−3m. PMID:5104982

  14. Fabrication of pullulan and pectin submicron fibers by electrospinning

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pullulan (PUL), a food grade polysaccharide, was fabricated into fibrous mats from fibers of submicron size by electrospinning. The effects of inorganic salts and polyanions present in the electrospinning solution on the properties of the resultant fibers was investigated. The inclusion of exogenous...

  15. Characterization of Pullulan/Chitosan Oligosaccharide/Montmorillonite Nanofibers Prepared by Electrospinning Technique.

    PubMed

    Rabbani, Mohammad Mahbub; Yang, Seong Baek; Park, Soo-Jin; Oh, Weontae; Yeum, Jeong Hyun

    2016-06-01

    Pullulan/Chitosan oligosaccharide (COS)/Montmorillonite (MMT) hybrid nanofibers were electrospun from their aqueous solution using different Pullulan/COS mass ratios and variable amounts of MMT. The effects of Pullulan/COS mass ratios and MMT contents on the morphologies and properties of PulluIan/COS/MMT hybrid nanofibers were investigated. The obtained nanofibers were characterized with field emission-scanning electron microscopy (FE-SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), thermo gravimetric analysis (TGA), and tensile strength measurement. The Pullulan/COS mass ratio and MMT contents significantly influence the morphologies and properties of the Pullulan/COS/MMT hybrid nanofibers. Higher Pullulan contents than COS contents forms uniform and bead free nanofibers. The addition of COS to Pullulan improves the thermal stability of Pullulan/COS blend nanofibers. The incorporation of MMT to the Pullulan/COS/MMT hybrid nanofibers increase their fiber diameter, improves their thermal stability and tensile strength. These morphological changes and property enhancement depend on the amount of MMT added. The XRD and TEM results suggest the coexistence of Pullulan, COS and MMT within polymer matrix through intercalation of polymer chain between silicate layers forming well-ordered multiplayer morphology with alternating polymeric and silicate layers. PMID:27427741

  16. Deoiledjatropha seed cake is a useful nutrient for pullulan production

    PubMed Central

    2012-01-01

    Background Ever increasing demand for fossil fuels is a major factor for rapid depletion of these non-renewable energy resources, which has enhanced the interest of finding out alternative sources of energy. In recent years jatropha seed oil has been used extensively for production of bio-diesel and has shown significant potential to replace petroleum fuels at least partially. De-oiled jatropha seed cake (DOJSC) which comprises of approximately 55 to 65% of the biomass is a byproduct of bio-diesel industry. DOJSC contains toxic components like phorbol esters which restricts its utilization as animal feed. Thus along with the enhancement of biodiesel production from jatropha, there is an associated problem of handling this toxic byproduct. Utilization of DOJSC as a feed stock for production of biochemicals may be an attractive solution to the problem. Pullulan is an industrially important polysaccharide with several potential applications in food, pharmaceuticals and cosmetic industries. However, the major bottleneck for commercial utilization of pullulan is its high cost. A cost effective process for pullulan production may be developed using DOJSC as sole nutrient source which will in turn also help in utilization of the byproduct of bio-diesel industry. Results In the present study, DOJSC has been used as a nutrient for production of pullulan, in place of conventional nutrients like yeast extract and peptone. Process optimization was done in shake flasks, and under optimized conditions (8% DOJSC, 15% dextrose, 28°C temperature, 200 rpm, 5% inoculum, 6.0 pH) 83.98 g/L pullulan was obtained. The process was further validated in a 5 L laboratory scale fermenter. Conclusion This is the first report of using DOJSC as nutrient for production of an exopolysaccharide. Successful use of DOJSC as nutrient will help in finding significant application of this toxic byproduct of biodiesel industry. This in turn also have a significant impact on cost reduction and may lead to

  17. 7 CFR 17.1 - General.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... practicable, under the authority of title I of the Agricultural Trade Development and Assistance Act of 1954... Secretary of Agriculture SALES OF AGRICULTURAL COMMODITIES MADE AVAILABLE UNDER TITLE I OF THE AGRICULTURAL TRADE DEVELOPMENT AND ASSISTANCE ACT OF 1954, AS AMENDED § 17.1 General. (a) What this part covers....

  18. Production of novel types of antibacterial liamocins by diverse strains of Aureobasidium pullulans grown on different culture media

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Objective: The objective was to compare production of antibacterial liamocins by diverse strains of A. pullulans grown on different culture media. Results: Liamocins produced by strains of A. pullulans have potential agricultural and pharmaceutical applications as antibacterials with specificity aga...

  19. Real-time and postprocessing holographic effects in dichromated pullulan.

    PubMed

    Savić, Svetlana; Pantelić, Dejan; Jakovijević, Dragica

    2002-08-01

    Experimental results concerning both real-time and postprocessing (after-development) behavior of a novel photosensitive material, dichromate-sensitized pullulan (DCP), are investigated. The exposure mechanism and possibilities for controlling holographic grating properties are discussed. We have shown that it is possible to maximize the diffraction efficiency of interference gratings after development by controlling diffraction efficiency in real time. Stronger real-time effects of DCP compared with those of dichromated gelatin are achieved. PMID:12153075

  20. Genetic Diversity of Tropical Isolates and Color Variants of Aureobasidium pullulans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The polymorphic fungus Aureobasidium pullulans was isolated from many provinces in tropical Thailand and cultured in pullulan production medium (PM) containing sucrose and peptone as carbon and nitrogen sources, respectively. Liquid PM cultures varied in color from cream to pale pink, light burgund...

  1. Aureobasidium pullulans as a biocontrol agent of postharvest pathogens of apples in Uruguay

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The yeast, Aureobasidium pullulans, was the microorganism most frequently recovered from the surface of apple fruit (cv. Red Delicious) stored in commercial cold chambers for six months. In the present work, ten isolates of Aureobasidium pullulans were assayed to determine if they could control blu...

  2. Phylogenetic classification of Aureobasidium pullulans strains for production of feruloyl esterase

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective was to phylogenetically classify diverse strains of A. pullulans and determine their production of feruloyl esterase. Seventeen strains from the A. pullulans literature were phylogenetically classified. Phenotypic traits of color variation and endo-ß-1,4-xylanase overproduction were as...

  3. Production of anti-streptococcal liamocins from agricultural biomass by Aureobasidium pullulans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Liamocins are unique heavier-than-water “oils” produced by certain strains of the fungus Aureobasidium pullulans. Liamocins have antibacterial activity with specificity for Streptococcus sp. Previous studies reported that liamocin yields were highest from strains of A. pullulans belonging to phylog...

  4. Rheological properties of pullulan-sodium alginate based solutions during film formation.

    PubMed

    Xiao, Qian; Tong, Qunyi; Zhou, Yujia; Deng, Fangming

    2015-10-01

    During film formation, the rheological properties of pullulan, sodium alginate, and blends, dried at 50°C were studied using an oscillatory rheometer. According to the drying curves, the drying process of pullulan, alginate, and blend films was divided into three stages. At the first drying stage, four samples exhibited typical liquid-like viscoelastic behavior. As the drying proceeded (polysaccharide concentration up to 75%), pure pullulan chains formed an entangled network, whereas coupling of alginate molecules gave a weak gel. At this drying stage, complex viscosity data for 75% alginate and blends were fitted with the power law equation. The effects of drying on the mechanical properties of pullulan-sodium alginate based samples were analyzed using the generalized Maxwell model, and their relaxation spectra were determined. The rheological properties during drying obtained from this study is essential for understanding film-forming mechanism and predicting the properties of pullulan-sodium alginate based edible films. PMID:26076600

  5. Effect of oligosaccharides derived from Laminaria japonica-incorporated pullulan coatings on preservation of cherry tomatoes.

    PubMed

    Wu, Shengjun; Lu, Mingsheng; Wang, Shujun

    2016-05-15

    Laminaria japonica-derived oligosaccharides (LJOs) exhibit antibacterial and antioxidant activities, and pullulan is a food thickener that can form impermeable films. The ability of pullulan coatings with various LJO concentrations (1% pullulan+0.1%, 0.2% or 0.3% LJOs) to preserve cherry tomatoes during storage at room temperature was investigated. The LJO-incorporated pullulan coatings were found to effectively reduce respiratory intensity, vitamin C loss, weight loss and softening, as well as to increase the amount of titratable acid and the overall likeness of fruit compared with the control. These effects were observed to be dose-dependent. Therefore, using LJO-incorporated pullulan coatings can extend the shelf life of cherry tomatoes.

  6. Effect of oligosaccharides derived from Laminaria japonica-incorporated pullulan coatings on preservation of cherry tomatoes.

    PubMed

    Wu, Shengjun; Lu, Mingsheng; Wang, Shujun

    2016-05-15

    Laminaria japonica-derived oligosaccharides (LJOs) exhibit antibacterial and antioxidant activities, and pullulan is a food thickener that can form impermeable films. The ability of pullulan coatings with various LJO concentrations (1% pullulan+0.1%, 0.2% or 0.3% LJOs) to preserve cherry tomatoes during storage at room temperature was investigated. The LJO-incorporated pullulan coatings were found to effectively reduce respiratory intensity, vitamin C loss, weight loss and softening, as well as to increase the amount of titratable acid and the overall likeness of fruit compared with the control. These effects were observed to be dose-dependent. Therefore, using LJO-incorporated pullulan coatings can extend the shelf life of cherry tomatoes. PMID:26775974

  7. Effect of pullulan on the water distribution, microstructure and textural properties of rice starch gels during cold storage.

    PubMed

    Chen, Long; Tian, Yaoqi; Tong, Qunyi; Zhang, Zipei; Jin, Zhengyu

    2017-01-01

    The effects of pullulan on the water distribution, microstructure and textural properties of rice starch gels during cold storage were investigated by low field-nuclear magnetic resonance (LF-NMR), scanning electron microscope (SEM), and texture profile analysis (TPA). The addition of pullulan reduced the transversal relaxation time of rice starch gels during cold storage. The microstructure of rice starch gel with 0.5% pullulan was denser and more uniform compared with that of rice starch without pullulan in each period of storage time. With regard to textural properties, 0.01% pullulan addition did not significantly change the texture of rice starch gels, while 0.5% pullulan addition appeared to reduce the hardness and retain the springiness of rice starch gels (P⩽0.05). The restriction effects of pullulan on water mobility and starch retrogradation were hypothesized to be mainly responsible for the water retention, gel structure maintenance, and modification of the textural attributes of rice starch gels.

  8. Hydrolysis of xylan by an immobilized xylanase from Aureobasidium pullulans

    SciTech Connect

    Allenza, P.; Scherl, D.S.; Detroy, R.W.; Leathers, T.D.; Scott, C.D. .

    1986-01-01

    The beta-(1,4)-linked xylose residues that comprise the backbone of the abundant plant polymer xylan can be released by enzymic hydrolysis. Xylanase, which is produced in exceptionally high levels by the color-variant strain Y-2311-1 of A. pullulans, was immobilized onto a macroporous ceramic carrier. Despite a low coupling efficiency, it was possible to run the reactor under a wide range of conditions with flow rates of 3-10 bed volumes/minute of 1% soluble xylan with no detectable leaching of enzyme. The size distribution of products and rate of xylan hydrolysis were very similar for the immobilized and soluble enzymes. (Refs. 13).

  9. Preservation of Brussels Sprouts by Pullulan Coating Containing Oregano Essential Oil.

    PubMed

    Kraśniewska, Karolina; Gniewosz, Małgorzata; Kosakowska, Olga; Cis, Aneta

    2016-03-01

    In this study, the effectiveness of pullulan (a fungal polysaccharide) film containing oregano essential oil (OEO) at 1.0 to 10.0% was evaluated against bacteria, yeasts, and molds. The quality of the sprouts, as determined by weight loss, color, and appearance, was monitored during storage at 2 and 16°C. An organoleptic evaluation of odor preference and odor acceptability of OEO on the Brussels sprouts was also conducted. The antimicrobial activity of pullulan films with OEO increased significantly with the increase in OEO concentration (1 to 10%). Pullulan films with OEO were more effective for inhibiting the growth of yeasts and molds than for inhibiting gram-positive and gram-negative bacteria. Pullulan with 1.0% OEO was an effective combination and was used subsequently as the base coating for maintaining the safety and quality of fresh Brussels sprouts stored at 16°C for 14 days. The pullulan coating containing 1.0% OEO reduced Aspergillus niger populations by 2 log CFU/g. This coating also reduced weight loss in the sprouts. Compared with uncoated Brussels sprouts, the percent weight loss after 14 days was reduced in samples coated with pullulan and with pullulan plus 1% OEO by 3.81 and 6.06%, respectively, after storage at 2°C and by 8.04 and 9.30%, respectively, after storage 16°C. The coating also significantly reduced changes in general appearance and color during storage. Evaluation of the organoleptic properties indicated that pullulan containing OEO had only a slight detrimental effect on odor properties. Incorporating OEO into a delivery system for antimicrobial compounds in pullulan coatings extended the microbiological shelf life of Brussels sprouts. PMID:26939662

  10. Esterases and putative lipases from tropical isolates of Aureobasidium pullulans.

    PubMed

    Kudanga, Tukayi; Mwenje, Eddie; Mandivenga, Faith; Read, John S

    2007-04-01

    Esterases and lipases have been studied in a number of fungi, though very little is known about esterases from Aureobasidium pullulans especially from the African tropics. In this study, forty-two Zimbabwean isolates were screened for lipase activity on tributyrin agar. Extracellular esterase activities of seven selected isolates were studied under varying conditions using para-nitrophenol acetate as substrate. Twenty isolates (48%) showed lipolytic activity; sixteen showed negative results for lipase activity while the rest showed weak activities. Esterase activities in broth cultures ranged from 0.011-0.223 mmol/microg protein/min while activities ranged from 1.5-12.8 U/ml under solid state fermentation. The esterases were optimally active at pH 7.6-8.0, showed a temperature optimum of 35 degrees C and retained more than 50% activity at temperatures up to 60 degrees C and at pH 4.0-7.0 after 150 min. Enzyme production was optimal after 5-6 days with diammonium hydrogen phosphate as nitrogen source. Isolates showed variations in preference for carbon source for esterase production. The A. pullulans esterases differed from most fungal esterases in that they are optimally active in alkaline conditions and are active over a broad pH range. PMID:17440916

  11. Effects of plastic composite support and pH profiles on pullulan production in a biofilm reactor.

    PubMed

    Cheng, Kuan-Chen; Demirci, Ali; Catchmark, Jeffrey M

    2010-04-01

    Pullulan is a linear homopolysaccharide which is composed of glucose units and often described as alpha-1, 6-linked maltotriose. The applications of pullulan range from usage as blood plasma substitutes to environmental pollution control agents. In this study, a biofilm reactor with plastic composite support (PCS) was evaluated for pullulan production using Aureobasidium pullulans. In test tube fermentations, PCS with soybean hulls, defatted soy bean flour, yeast extract, dried bovine red blood cells, and mineral salts was selected for biofilm reactor fermentation (due to its high nitrogen content, moderate nitrogen leaching rate, and high biomass attachment). Three pH profiles were later applied to evaluate their effects on pullulan production in a PCS biofilm reactor. The results demonstrated that when a constant pH at 5.0 was applied, the time course of pullulan production was advanced and the concentration of pullulan reached 32.9 g/L after 7-day cultivation, which is 1.8-fold higher than its respective suspension culture. The quality analysis demonstrated that the purity of produced pullulan was 95.8% and its viscosity was 2.4 centipoise. Fourier transform infrared spectroscopy spectra also supported the supposition that the produced exopolysaccharide was mostly pullulan. Overall, this study demonstrated that a biofilm reactor can be successfully implemented to enhance pullulan production and maintain its high purity.

  12. Protection of Lactobacillus acidophilus NRRL-B 4495 under in vitro gastrointestinal conditions with whey protein/pullulan microcapsules.

    PubMed

    Çabuk, Burcu; Tellioğlu Harsa, Şebnem

    2015-12-01

    In this research, whey protein/pullulan (WP/pullulan) microcapsules were developed in order to assess its protective effect on the viability of Lactobacillus acidophilus NRRL-B 4495 under in vitro gastrointestinal conditions. Results demonstrated that WP/pullulan microencapsulated cells exhibited significantly (p ≤ 0.05) higher resistance to simulated gastric acid and bile salt. Pullulan incorporation into protein wall matrix resulted in improved survival as compared to free cells after 3 h incubation in simulated gastric solution. Moreover WP/pullulan microcapsules were found to release over 70% of encapsulated L. acidophilus NRRL-B 4495 cells within 1 h. The effect of encapsulation during refrigerated storage was also studied. Free bacteria exhibited 3.96 log reduction while, WP/pullulan encapsulated bacteria showed 1.64 log reduction after 4 weeks of storage. PMID:26100319

  13. Protection of Lactobacillus acidophilus NRRL-B 4495 under in vitro gastrointestinal conditions with whey protein/pullulan microcapsules.

    PubMed

    Çabuk, Burcu; Tellioğlu Harsa, Şebnem

    2015-12-01

    In this research, whey protein/pullulan (WP/pullulan) microcapsules were developed in order to assess its protective effect on the viability of Lactobacillus acidophilus NRRL-B 4495 under in vitro gastrointestinal conditions. Results demonstrated that WP/pullulan microencapsulated cells exhibited significantly (p ≤ 0.05) higher resistance to simulated gastric acid and bile salt. Pullulan incorporation into protein wall matrix resulted in improved survival as compared to free cells after 3 h incubation in simulated gastric solution. Moreover WP/pullulan microcapsules were found to release over 70% of encapsulated L. acidophilus NRRL-B 4495 cells within 1 h. The effect of encapsulation during refrigerated storage was also studied. Free bacteria exhibited 3.96 log reduction while, WP/pullulan encapsulated bacteria showed 1.64 log reduction after 4 weeks of storage.

  14. 46 CFR 34.17-1 - Application-T/ALL.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 1 2010-10-01 2010-10-01 false Application-T/ALL. 34.17-1 Section 34.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY TANK VESSELS FIREFIGHTING EQUIPMENT Fixed Foam Extinguishing Systems, Details § 34.17-1 Application—T/ALL. (a) Where a fixed foam extinguishing system...

  15. 46 CFR 96.17-1 - When required.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 4 2013-10-01 2013-10-01 false When required. 96.17-1 Section 96.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CARGO AND MISCELLANEOUS VESSELS VESSEL CONTROL AND MISCELLANEOUS SYSTEMS AND EQUIPMENT Magnetic Compass and Gyrocompass § 96.17-1 When required. (a) All mechanically propelled vessels in ocean...

  16. 46 CFR 96.17-1 - When required.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 4 2012-10-01 2012-10-01 false When required. 96.17-1 Section 96.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CARGO AND MISCELLANEOUS VESSELS VESSEL CONTROL AND MISCELLANEOUS SYSTEMS AND EQUIPMENT Magnetic Compass and Gyrocompass § 96.17-1 When required. (a) All mechanically propelled vessels in ocean...

  17. 46 CFR 34.17-1 - Application-T/ALL.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 1 2013-10-01 2013-10-01 false Application-T/ALL. 34.17-1 Section 34.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY TANK VESSELS FIREFIGHTING EQUIPMENT Fixed Foam Extinguishing Systems, Details § 34.17-1 Application—T/ALL. (a) Where a fixed foam extinguishing system...

  18. 46 CFR 34.17-1 - Application-T/ALL.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 1 2014-10-01 2014-10-01 false Application-T/ALL. 34.17-1 Section 34.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY TANK VESSELS FIREFIGHTING EQUIPMENT Fixed Foam Extinguishing Systems, Details § 34.17-1 Application—T/ALL. (a) Where a fixed foam extinguishing system...

  19. 46 CFR 34.17-1 - Application-T/ALL.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 1 2012-10-01 2012-10-01 false Application-T/ALL. 34.17-1 Section 34.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY TANK VESSELS FIREFIGHTING EQUIPMENT Fixed Foam Extinguishing Systems, Details § 34.17-1 Application—T/ALL. (a) Where a fixed foam extinguishing system...

  20. 46 CFR 34.17-1 - Application-T/ALL.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 1 2011-10-01 2011-10-01 false Application-T/ALL. 34.17-1 Section 34.17-1 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY TANK VESSELS FIREFIGHTING EQUIPMENT Fixed Foam Extinguishing Systems, Details § 34.17-1 Application—T/ALL. (a) Where a fixed foam extinguishing system...

  1. Emergence of Aureobasidium pullulans as human fungal pathogen and molecular assay for future medical diagnosis.

    PubMed

    Chan, Giek Far; Puad, Mohamad Safwan Ahmad; Chin, Chai Fung; Rashid, Noor Aini Abdul

    2011-09-01

    Despite the great importance of Aureobasidium pullulans in biotechnology, the fungus had emerged as an opportunistic human pathogen, especially among immunocompromised patients. Clinical detection of this rare human fungal pathogen presently relies on morphology diagnosis which may be misleading. Thus, a sensitive and accurate quantitative molecular assay for A. pullulans remains lacking. In this study, we presented the microscopy observations of A. pullulans that reveals the phenotypic plasticity of the fungus. A. pullulans-specific primers and molecular beacon probes were designed based on the fungal 18S ribosomal RNA (rRNA) gene. Comparison of two probes with varied quencher chemistry, namely BHQ-1 and Tamra, revealed high amplification efficiency of 104% and 108%, respectively. The optimized quantitative real-time PCR (qPCR) assays could detect and quantify up to 1 pg concentration of A. pullulans DNA. Both assays displayed satisfactory performance parameters at fast thermal cycling mode. The molecular assay has great potential as a molecular diagnosis tool for early detection of fungal infection caused by A. pullulans, which merits future study in clinical diagnosis.

  2. Statistical studies on high molecular weight pullulan production in solid state fermentation using jack fruit seed.

    PubMed

    Sugumaran, K R; Sindhu, R V; Sukanya, S; Aiswarya, N; Ponnusami, V

    2013-10-15

    The purpose of the work was to optimize the medium variables for maximizing pullulan production using jack fruit seed as a low cost substrate by Aureobasidium pullulans in solid state fermentation. Effects of K2HPO4, KH2PO4, ZnSO4·5H2O, MgSO4·7H2O, NaCl, (NH4)2SO4·5H2O, yeast extract, moisture content (%, w/w) in the production medium on pullulan production were studied using Plackett-Burman design. Production of pullulan was significantly affected by the medium variables namely KH2PO4, ZnSO4·5H2O, NaCl and moisture content (%, w/w). Then screened variables were optimized by Box Behnken experiment design. The pullulan obtained was characterized and confirmed by FTIR, (1)H NMR and (13)C NMR. Molecular weight of pullulan was found to be 1.733×10(6) g/mol by gel permeation chromatography (GPC).

  3. [Effects of sperminated pullulans on the pulmonary absorption of insulin].

    PubMed

    Seki, Toshinobu; Fukushi, Nanako; Maru, Hiroto; Kimura, Soichiro; Chono, Sumio; Egawa, Yuya; Morimoto, Kazuhiro; Ueda, Hideo; Morimoto, Yasunori

    2011-02-01

    Sperminated pullulans (SP) having different molecular weights (MWs) were prepared, and the enhancing effect on the pulmonary absorption of insulin in rats was examined. SP acted as enhancers of insulin absorption when a 0.1% solution was applied with insulin simultaneously and their enhancing effects depended on the MW of the SP; the same solutions exhibited low toxicity in the in vivo LDH leaching test. In the in vitro experiments using Calu-3 cells, tight junction-opening effects and a toxic effect of SP in the MTT assay were observed at lower concentrations compared with the in vivo experiments. A mucus layer might interfere with the interaction between SP and the cell surface and might suppress both these effects and toxicity. SP having a high MW will be useful for preparing safe and efficient formulations of peptide and protein drugs. The change in the localization of the tight junction proteins may be related to the permeation-enhancing mechanism of SP.

  4. Deterioration of expanded polystyrene caused by Aureobasidium pullulans var. melanogenum.

    PubMed

    Castiglia, Valeria C; Kuhar, Francisco

    2015-01-01

    An expanded-polystyrene factory located in northern Buenos Aires reported unusual dark spots causing esthetic damage in their production. A fungal strain forming black-olive colonies on extract malt agar medium was isolated from the damaged material and identified as Aureobasidium pullullans var. melanogenum. This fungus is particularly known for its capacity to produce hydrolytic enzymes and a biodegradable extracellular polysaccharide known as pullulan, which is used in the manufacture of packaging material for food and medicine. Laboratory tests were conducted to characterize its growth parameters. It was found that the organism was resistant to a wide range of pHs but did not survive at temperatures over 65°C. The proposed action plan includes drying of the material prior to packaging and disinfection of the machinery used in the manufacturing process and of the silos used for raw material storage. PMID:26165967

  5. Cellularized Bilayer Pullulan-Gelatin Hydrogel for Skin Regeneration.

    PubMed

    Nicholas, Mathew N; Jeschke, Marc G; Amini-Nik, Saeid

    2016-05-01

    Skin substitutes significantly reduce the morbidity and mortality of patients with burn injuries and chronic wounds. However, current skin substitutes have disadvantages related to high costs and inadequate skin regeneration due to highly inflammatory wounds. Thus, new skin substitutes are needed. By combining two polymers, pullulan, an inexpensive polysaccharide with antioxidant properties, and gelatin, a derivative of collagen with high water absorbency, we created a novel inexpensive hydrogel-named PG-1 for "pullulan-gelatin first generation hydrogel"-suitable for skin substitutes. After incorporating human fibroblasts and keratinocytes onto PG-1 using centrifugation over 5 days, we created a cellularized bilayer skin substitute. Cellularized PG-1 was compared to acellular PG-1 and no hydrogel (control) in vivo in a mouse excisional skin biopsy model using newly developed dome inserts to house the skin substitutes and prevent mouse skin contraction during wound healing. PG-1 had an average pore size of 61.69 μm with an ideal elastic modulus, swelling behavior, and biodegradability for use as a hydrogel for skin substitutes. Excellent skin cell viability, proliferation, differentiation, and morphology were visualized through live/dead assays, 5-bromo-2'-deoxyuridine proliferation assays, and confocal microscopy. Trichrome and immunohistochemical staining of excisional wounds treated with the cellularized skin substitute revealed thicker newly formed skin with a higher proportion of actively proliferating cells and incorporation of human cells compared to acellular PG-1 or control. Excisional wounds treated with acellular or cellularized hydrogels showed significantly less macrophage infiltration and increased angiogenesis 14 days post skin biopsy compared to control. These results show that PG-1 has ideal mechanical characteristics and allows ideal cellular characteristics. In vivo evidence suggests that cellularized PG-1 promotes skin regeneration and may

  6. Properties of edible films based on pullulan-chitosan blended film-forming solutions at different pH

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Influences of solution pH on the properties of pullulan-chitosan blended (Pul-Chi) films and the rheological properties of film-forming solutions were investigated. The extended conformation of chitosan in pH 4.0 solution increased intermolecular interactions with pullulan compared to the more compa...

  7. Modification of the mannitol biosynthetic pathway in Aureobasidium pullulans to alter the structure of the polyol lipid liamocin

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aureobasidium pullulans is an important industrial organism that is utilized for the production of numerous bioproducts, such as pullulan, ß-glucan, aureobasidin, and polymalic acid. Our laboratory is also interested in production of the extracellular polyol lipids, called liamocins, produced by cer...

  8. Polyols, not sugars, determine the structural diversity of anti-streptococcal liamocins produced by Aureobasidium pullulans strain NRRL 50380

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Liamocins are polyol-lipids produced by the fungus Aureobasidium pullulans, and have selective antibacterial activity against Streptococcus species. Liamocins produced by A. pullulans strain NRRL 50380 on sucrose medium have a D-mannitol head-group ester linked to 3,5-dihydroxydecanoate acyl chains,...

  9. Green synthesis of the silver nanoparticles mediated by pullulan and 6-carboxypullulan.

    PubMed

    Coseri, Sergiu; Spatareanu, Alina; Sacarescu, Liviu; Rimbu, Cristina; Suteu, Daniela; Spirk, Stefan; Harabagiu, Valeria

    2015-02-13

    Unoxidized and carboxylated pullulan (obtained by pullulan oxidation using TEMPO-sodium hypochlorite-sodium bromide system) have been used as mediators for the silver nanoparticles formation (AgNPs), under environment-friendly conditions: using aqueous solutions, room temperature and notably, by using both mediators as reducing and stabilizing agents. The formation of AgNPs was first screened by measuring the surface plasmon resonance peak in the range of 380-440 nm using UV-vis spectroscopy. The morphology of the synthesized silver nanoparticles was determined by TEM, which indicated that the AgNPs differ on shape and thickness of the polymer shell by varying the silver nitrate concentration, different size and shape of AgNPs was achieved. The presence of elemental silver and the crystalline structure of the AgNPs were confirmed by EDX and XRD analyses. Moreover, the possible functional groups of pullulan (oxidized pullulan) responsible for the reduction and stabilization of AgNPs were evaluated using FT-IR. The results showed that both, pullulan and 6-carboxypullulan could be successfully used as reducing as well as capping agents for the AgNPs synthesis which shows potential antimicrobial activity against Gram positive and Gram negative bacteria.

  10. Ketoprofen-loaded pomegranate seed oil nanoemulsion stabilized by pullulan: Selective antiglioma formulation for intravenous administration.

    PubMed

    Ferreira, Luana M; Cervi, Verônica F; Gehrcke, Mailine; da Silveira, Elita F; Azambuja, Juliana H; Braganhol, Elizandra; Sari, Marcel H M; Zborowski, Vanessa A; Nogueira, Cristina W; Cruz, Letícia

    2015-06-01

    This study aimed to prepare pomegranate seed oil nanoemulsions containing ketoprofen using pullulan as a polymeric stabilizer, and to evaluate antitumor activity against in vitro glioma cells. Formulations were prepared by the spontaneous emulsification method and different concentrations of pullulan were tested. Nanoemulsions presented adequate droplet size, polydispersity index, zeta potential, pH, ketoprofen content and encapsulation efficiency. Nanoemulsions were able to delay the photodegradation profile of ketoprofen under UVC radiation, regardless of the concentration of pullulan. In vitro release study indicates that nanoemulsions were able to release approximately 95.0% of ketoprofen in 5h. Free ketoprofen and formulations were considered hemocompatible at 1 μg/mL, in a hemolysis study, for intravenous administration. In addition, a formulation containing the highest concentration of pullulan was tested against C6 cell line and demonstrated significant activity, and did not reduce fibroblasts viability. Thus, pullulan can be considered an interesting excipient to prepare nanostructured systems and nanoemulsion formulations can be considered promising alternatives for the treatment of glioma.

  11. Development of chitosan-pullulan composite nanoparticles for nasal delivery of vaccines: in vivo studies.

    PubMed

    Cevher, Erdal; Salomon, Stefan K; Somavarapu, Satyanarayana; Brocchini, Steve; Alpar, H Oya

    2015-01-01

    Here, we aimed at developing chitosan/pullulan composite nanoparticles and testing their potential as novel systems for the nasal delivery of diphtheria toxoid (DT). All the chitosan derivatives [N-trimethyl (TMC), chloride and glutamate] and carboxymethyl pullulan (CMP) were synthesised and antigen-loaded composites were prepared by polyion complexation of chitosan and pullulan derivatives (particle size: 239-405 nm; surface charge: +18 and +27 mV). Their immunological effects after intranasal administration to mice were compared to intramuscular route. Composite nanoparticles induced higher levels of IgG responses than particles formed with chitosan derivative and antigen. Nasally administered TMC-pullulan composites showed higher DT serum IgG titre when compared with the other composites. Co-encapsulation of CpG ODN within TMC-CMP-DT nanoparticles resulted in a balanced Th1/Th2 response. TMC/pullulan composite nanoparticles also induced highest cytokine levels compared to those of chitosan salts. These findings demonstrated that TMC-CMP-DT composite nanoparticles are promising delivery system for nasal vaccination. PMID:26480962

  12. 1 CFR 17.1 - Receipt and processing.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 1 General Provisions 1 2014-01-01 2012-01-01 true Receipt and processing. 17.1 Section 17.1 General Provisions ADMINISTRATIVE COMMITTEE OF THE FEDERAL REGISTER PREPARATION, TRANSMITTAL, AND PROCESSING OF DOCUMENTS FILING FOR PUBLIC INSPECTION AND PUBLICATION SCHEDULES Receipt and Processing §...

  13. 1 CFR 17.1 - Receipt and processing.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 1 General Provisions 1 2012-01-01 2012-01-01 false Receipt and processing. 17.1 Section 17.1 General Provisions ADMINISTRATIVE COMMITTEE OF THE FEDERAL REGISTER PREPARATION, TRANSMITTAL, AND PROCESSING OF DOCUMENTS FILING FOR PUBLIC INSPECTION AND PUBLICATION SCHEDULES Receipt and Processing §...

  14. 1 CFR 17.1 - Receipt and processing.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 1 General Provisions 1 2013-01-01 2012-01-01 true Receipt and processing. 17.1 Section 17.1 General Provisions ADMINISTRATIVE COMMITTEE OF THE FEDERAL REGISTER PREPARATION, TRANSMITTAL, AND PROCESSING OF DOCUMENTS FILING FOR PUBLIC INSPECTION AND PUBLICATION SCHEDULES Receipt and Processing §...

  15. 1 CFR 17.1 - Receipt and processing.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 1 General Provisions 1 2011-01-01 2011-01-01 false Receipt and processing. 17.1 Section 17.1 General Provisions ADMINISTRATIVE COMMITTEE OF THE FEDERAL REGISTER PREPARATION, TRANSMITTAL, AND PROCESSING OF DOCUMENTS FILING FOR PUBLIC INSPECTION AND PUBLICATION SCHEDULES Receipt and Processing §...

  16. 1 CFR 17.1 - Receipt and processing.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 1 General Provisions 1 2010-01-01 2010-01-01 false Receipt and processing. 17.1 Section 17.1 General Provisions ADMINISTRATIVE COMMITTEE OF THE FEDERAL REGISTER PREPARATION, TRANSMITTAL, AND PROCESSING OF DOCUMENTS FILING FOR PUBLIC INSPECTION AND PUBLICATION SCHEDULES Receipt and Processing §...

  17. Exceptional oxygen barrier performance of pullulan nanocomposites with ultra-low loading of graphene oxide

    NASA Astrophysics Data System (ADS)

    Uysal Unalan, Ilke; Wan, Chaoying; Figiel, Łukasz; Olsson, Richard T.; Trabattoni, Silvia; Farris, Stefano

    2015-07-01

    Polymer nanocomposites are increasingly important in food packaging sectors. Biopolymer pullulan is promising in manufacturing packaging films or coatings due to its excellent optical clarity, mechanical strength, and high water-solubility as compared to other biopolymers. This work aims to enhance its oxygen barrier properties and overcome its intrinsic brittleness by utilizing two-dimensional planar graphene oxide (GO) nanoplatelets. It has been found that the addition of only 0.2 wt% of GO enhanced the tensile strength, Young’s modulus, and elongation at break of pullulan films by about 40, 44 and 52%, respectively. The light transmittance at 550 nm of the pullulan/GO films was 92.3% and haze values were within 3.0% threshold, which meets the general requirement for food packaging materials. In particular, the oxygen permeability coefficient of pullulan was reduced from 6337 to 2614 mL μm m-2 (24 h-1) atm-1 with as low as 0.05 wt% of GO loading and further to 1357 mL μm m-2 (24 h-1) atm-1 when GO concentration reached 0.3 wt%. The simultaneous improvement of the mechanical and oxygen barrier properties of pullulan was ascribed to the homogeneous distribution and prevalent unidirectional alignment of GO nanosheets, as determined from the characterization and theoretical modelling results. The exceptional oxygen barrier properties of pullulan/GO nanocomposites with enhanced mechanical flexibility and good optical clarity will add new values to high performance food packaging materials.

  18. Effect of pullulan on the water distribution, microstructure and textural properties of rice starch gels during cold storage.

    PubMed

    Chen, Long; Tian, Yaoqi; Tong, Qunyi; Zhang, Zipei; Jin, Zhengyu

    2017-01-01

    The effects of pullulan on the water distribution, microstructure and textural properties of rice starch gels during cold storage were investigated by low field-nuclear magnetic resonance (LF-NMR), scanning electron microscope (SEM), and texture profile analysis (TPA). The addition of pullulan reduced the transversal relaxation time of rice starch gels during cold storage. The microstructure of rice starch gel with 0.5% pullulan was denser and more uniform compared with that of rice starch without pullulan in each period of storage time. With regard to textural properties, 0.01% pullulan addition did not significantly change the texture of rice starch gels, while 0.5% pullulan addition appeared to reduce the hardness and retain the springiness of rice starch gels (P⩽0.05). The restriction effects of pullulan on water mobility and starch retrogradation were hypothesized to be mainly responsible for the water retention, gel structure maintenance, and modification of the textural attributes of rice starch gels. PMID:27507528

  19. Antibacterial activity of liamocins oil from Aureobasidium pullulans is specific for species of Streptococcus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Liamocins are a heterogeneous mixture of denser-than-water oils produced by the fungus Aureobasidium pullulans. Liamocins have unique chemical structures with a mannitol head group linked to long chain polyester tails consisting of multiple 3,5-dihydroxydecanoic acid ester groups, some of which are ...

  20. Poly(beta-L-malic acid) from agricultural substrates by Aureobasidium pullulans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We report here for the first time the production of poly(beta-L-malic acid) (PMA) from agricultural substrates by the yeastlike fungus Aureobasidium pullulans. PMA is a natural biopolyester that has primarily been studied for biomedical uses as a drug carrier. However, PMA also has potential as a ...

  1. Liamocin oil from Aureobasidium pullulans has antibacterial activity with specificity for species of Streptococcus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Liamocin oil from Aureobasidium pullulans NRRL 50380 was tested for antibacterial activity. Liamocins inhibited growth of Streptococcus agalactiae, S. uberis, S. mitis, S. infantarius, and S. mutans, with minimum inhibitory concentrations from 20 'g/ml to 78 'g/ml. Enterococcus faecalis was less sus...

  2. Bioproducts and morphological features of diverse isolates of the fungus Aureobasidium pullulans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aureobasidium pullulans is a fungus included among the “black yeasts.” Although many strains are predominantly yeast-like, the species is actually polymorphic, exhibiting a variety of complex forms. The fungus is ubiquitous, routinely found on the surface of leaves, wood, painted walls, etc. We rece...

  3. QUANTITATIVE IMAGING AND STATISTICAL ANALYSIS OF FLUORESCENCE IN SITU HYBRIDIZATION (FISH) OF AUREOBASIDIUM PULLULANS. (R823845)

    EPA Science Inventory

    Abstract

    Image and multifactorial statistical analyses were used to evaluate the intensity of fluorescence signal from cells of three strains of A. pullulans and one strain of Rhodosporidium toruloides, as an outgroup, hybridized with either a universal o...

  4. Characteristics, protein engineering and applications of microbial thermostable pullulanases and pullulan hydrolases.

    PubMed

    Nisha, M; Satyanarayana, T

    2016-07-01

    Pullulan hydrolyzing enzymes are endoacting, classified based on the substrate specificity and hydrolysis products as pullulanases (type I and II) and pullulan hydrolases (type I, II and III). Pullulanases and pullulan hydrolase type I are produced by bacteria and archaea. Among bacteria, many mesophilic, thermophilic and hyperthermophilic bacteria produce pullulanases and neopullulanases. While pullulan hydrolase type II and type III are produced by fungi and archaea, respectively. These are multi-domain proteins with three conserved catalytic acidic residues of the glycosyl hydrolases. The recent advances in molecular biology and protein engineering via mutagenesis and truncation led to improvement in thermostability, catalytic activity and substrate specificity. Pullulanases are debranching enzymes, which are widely employed in starch saccharification that minimizes the use of glucoamylase (approx. 50 %) and reduces the total reaction time of the industrial starch conversion process. The thermostable amylopullulanases are useful in one-step starch liquefaction and saccharification, which replaces amylolytic enzymes like α-amylase and glucoamylase, thus resulting in the reduction in the cost of sugar production. The enzymes also find application in making resistant starches and as an antistale in bread making. Panose and isopanose containing syrups are useful as prebiotics, while panose has also been reported to display anticarcinogenic activity. This review focuses on the distinguishing features of these enzymes based on the analysis of amino acid sequences and domain structure, besides highlighting recent advances in the molecular biology and protein engineering for enhancing their thermostability, catalytic activity and substrate specificity. This review also briefly summarizes the potential applications of pullulanases and pullulan hydrolases. PMID:27142298

  5. 46 CFR 96.17-1 - When required.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... MISCELLANEOUS SYSTEMS AND EQUIPMENT Magnetic Compass and Gyrocompass § 96.17-1 When required. (a) All mechanically propelled vessels in ocean or coastwise service must be fitted with a magnetic compass. (b) All... gyrocompass in addition to the magnetic compass. (c) Each vessel must have an illuminated repeater for...

  6. 46 CFR 96.17-1 - When required.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... MISCELLANEOUS SYSTEMS AND EQUIPMENT Magnetic Compass and Gyrocompass § 96.17-1 When required. (a) All mechanically propelled vessels in ocean or coastwise service must be fitted with a magnetic compass. (b) All... gyrocompass in addition to the magnetic compass. (c) Each vessel must have an illuminated repeater for...

  7. 46 CFR 195.17-1 - When required.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... MISCELLANEOUS SYSTEMS AND EQUIPMENT Radar § 195.17-1 When required. All mechanically propelled vessels of 1,600 gross tons and over in ocean or coastwise service must be fitted with a marine radar system for surface navigation. Facilities for plotting radar readings must be provided on the bridge....

  8. 46 CFR 195.17-1 - When required.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... MISCELLANEOUS SYSTEMS AND EQUIPMENT Radar § 195.17-1 When required. All mechanically propelled vessels of 1,600 gross tons and over in ocean or coastwise service must be fitted with a marine radar system for surface navigation. Facilities for plotting radar readings must be provided on the bridge....

  9. 46 CFR 195.17-1 - When required.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... MISCELLANEOUS SYSTEMS AND EQUIPMENT Radar § 195.17-1 When required. All mechanically propelled vessels of 1,600 gross tons and over in ocean or coastwise service must be fitted with a marine radar system for surface navigation. Facilities for plotting radar readings must be provided on the bridge....

  10. 46 CFR 195.17-1 - When required.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... MISCELLANEOUS SYSTEMS AND EQUIPMENT Radar § 195.17-1 When required. All mechanically propelled vessels of 1,600 gross tons and over in ocean or coastwise service must be fitted with a marine radar system for surface navigation. Facilities for plotting radar readings must be provided on the bridge....

  11. 46 CFR 195.17-1 - When required.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... MISCELLANEOUS SYSTEMS AND EQUIPMENT Radar § 195.17-1 When required. All mechanically propelled vessels of 1,600 gross tons and over in ocean or coastwise service must be fitted with a marine radar system for surface navigation. Facilities for plotting radar readings must be provided on the bridge....

  12. 46 CFR 96.17-1 - When required.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... MISCELLANEOUS SYSTEMS AND EQUIPMENT Magnetic Compass and Gyrocompass § 96.17-1 When required. (a) All mechanically propelled vessels in ocean or coastwise service must be fitted with a magnetic compass. (b) All... gyrocompass in addition to the magnetic compass. (c) Each vessel must have an illuminated repeater for...

  13. 40 CFR 17.1 - Purpose of these rules.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ....1 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY GENERAL IMPLEMENTATION OF THE EQUAL ACCESS TO JUSTICE ACT IN EPA ADMINISTRATIVE PROCEEDINGS General Provisions § 17.1 Purpose of these rules...) of the Equal Access to Justice Act, Public Law No. 96-481. Under the Act, an eligible party...

  14. 40 CFR 17.1 - Purpose of these rules.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ....1 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY GENERAL IMPLEMENTATION OF THE EQUAL ACCESS TO JUSTICE ACT IN EPA ADMINISTRATIVE PROCEEDINGS General Provisions § 17.1 Purpose of these rules...) of the Equal Access to Justice Act, Public Law No. 96-481. Under the Act, an eligible party...

  15. 40 CFR 17.1 - Purpose of these rules.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ....1 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY GENERAL IMPLEMENTATION OF THE EQUAL ACCESS TO JUSTICE ACT IN EPA ADMINISTRATIVE PROCEEDINGS General Provisions § 17.1 Purpose of these rules...) of the Equal Access to Justice Act, Public Law No. 96-481. Under the Act, an eligible party...

  16. 40 CFR 17.1 - Purpose of these rules.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ....1 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY GENERAL IMPLEMENTATION OF THE EQUAL ACCESS TO JUSTICE ACT IN EPA ADMINISTRATIVE PROCEEDINGS General Provisions § 17.1 Purpose of these rules...) of the Equal Access to Justice Act, Public Law No. 96-481. Under the Act, an eligible party...

  17. 40 CFR 17.1 - Purpose of these rules.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ....1 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY GENERAL IMPLEMENTATION OF THE EQUAL ACCESS TO JUSTICE ACT IN EPA ADMINISTRATIVE PROCEEDINGS General Provisions § 17.1 Purpose of these rules...) of the Equal Access to Justice Act, Public Law No. 96-481. Under the Act, an eligible party...

  18. Antifungal activity of transparent nanocomposite thin films of pullulan and silver against Aspergillus niger.

    PubMed

    Pinto, Ricardo J B; Almeida, Adelaide; Fernandes, Susana C M; Freire, Carmen S R; Silvestre, Armando J D; Neto, Carlos Pascoal; Trindade, Tito

    2013-03-01

    Silver has been mainly investigated as an antibacterial agent and less as a fungicide in which concerns antimicrobial properties. In this research, the antifungal activity of composite films of pullulan and Ag nanoparticles (NP) against Aspergillus niger was evaluated using standard protocols. These new materials were prepared as transparent cast films (66-74 μm thickness) from Ag hydrosols containing the polysaccharide. Fungal growth inhibition was observed in the presence of such silver nanocomposite films. Moreover, disruption of the spores cells of A. niger was probed for the first time by means of scanning electron microscopy (SEM). This effect occurred in the presence of the nanocomposites due to Ag NP dispersed as fillers in pullulan. This polysaccharide was used here as a biocompatible matrix, hence making these nanocomposites beneficial for the development of antifungal packaging materials.

  19. Pullulan-protamine as efficient haemocompatible gene delivery vector: synthesis and in vitro characterization.

    PubMed

    Priya, S S; Rekha, M R; Sharma, Chandra P

    2014-02-15

    Biodegradable non-viral vectors with good transfection efficiency is essential for successful gene delivery. The purpose of this study was to design a non-viral vector by conjugating protamine to pullulan and elucidate the potential use of pullulan protamine conjugate (PPA) as an effective, non toxic and haemocompatible gene delivery system. The particle size and surface charge were measured using Nanosizer. Derivatization was confirmed by NMR, FTIR and DSC analyses. Acid base titration revealed the buffering behaviour of the conjugate. The protection of DNA from nuclease enzyme and interaction of plasma components on the stability of nanoplexes were also analysed. The uptake studies confirmed the plasmid delivery into the nucleus and the inhibitor studies determined the uptake mechanism. Transfection experiments revealed the capability of PPA to cellular uptake in C6 cells and facilitate high gene expression. Thus, PPA proves to be a promising non-viral vector.

  20. LX-17-1 Stockpile Returned Material Lot Comparison

    SciTech Connect

    Gagliardi, F.; Pease, S.; Willey, T.

    2015-02-18

    Many different lots of LX-17 have been produced over the years. Two varieties of LX-17, LX-17-0 and LX-17-1, have at one point or another been a part of the Livermore stockpile systems. LX-17-0 was made with dry-aminated TATB whereas LX-17-1 was made with wet-aminated TATB. Both versions have the same TATB to Kel-F 800 mass ratio of 92.5%/7.5%. Both kinds of LX-17 were formulated at Holston during the late 1970s or early to mid-1980s and were certified to have met the necessary specifications that cover the purity, particle size range, explosive to binder ratio, etc. In recent years, Trevor Willy and others have performed a detailed evaluation of solid parts made from each of the LX-17 lots manufactured at Holston. Using the Advanced Light Source at LBNL, Willey and his colleagues radiographed many samples from isostatic pressings using the same scanning conditions. In their investigation they identified that even though the bulk composition can be the same, there may exist a large spread in how smoothly the TATB and binder were distributed within the radiographed volume of different lots of material.1 Overall, the dry-aminated TATB-based material, LX-17-0, had a smooth TATB and binder distribution, whereas the wet-aminated TATB-based LX-17-1 showed a wide range of binder distributions. The results for five different LX-17-1 lots are shown in Figure 1. The wide variation in material distribution has raised the question about whether or not this sort variability will cause significant differences in mechanical behavior.

  1. Improving functionality of whole egg powder by the addition of gelatine, lactose, and pullulan.

    PubMed

    Koç, Mehmet; Koç, Banu; Susyal, Gonca; Yilmazer, Melike Sakin; Bağdatlıoğlu, Neriman; Kaymak-Ertekin, Figen

    2011-01-01

    The addition of gelatine (G), lactose (L), pullulan (P), and their mixtures at equal ratios (gelatine + lactose [GL] and gelatin + pullulan [GP]) to whole egg prior to drying was studied. Their effects on the functional properties (emulsion and gelling ability, water holding capacity of gel, foaming, color) and soluble protein content of spray dried egg powder during the 6 mo of storage at 20 °C and 50% relative humidity were investigated. It was demonstrated that the emulsion and foaming stability, water holding capacity of gel, and color change were significantly affected by the storage time, whereas storage time did not affect the strength of gel texture prepared by egg powders. Gelatine and pullulan improved the foaming stability and water holding capacity. Lactose caused a decrease in emulsion and foaming stability values. The maximum color change was observed for the plain egg powder, showing that mixing whole egg with carbohydrate- and/or protein-based additives before the drying process preserved the color of egg powder. Adding carbohydrate and/or protein caused significant changes in functional properties of egg powder. PMID:22416722

  2. Printed paper sensors for serum lactate dehydrogenase using pullulan-based inks to immobilize reagents.

    PubMed

    Kannan, Balamurali; Jahanshahi-Anbuhi, Sana; Pelton, Robert H; Li, Yingfu; Filipe, Carlos D M; Brennan, John D

    2015-09-15

    In this study, a paper-based point-of-care (POC) colorimetric biosensor was developed for the detection of lactate dehydrogenase in serum using a nonporous, oxygen impermeable reversibly gelling polysaccharide material based on pullulan. The pullulan could be printed onto paper surfaces along with all required assay reagents, providing a means for high-stability immobilization of all reagents on paper. Serum containing lactate dehydrogenase (LDH) was directly spotted on to the pullulan-coated bioactive paper and provided quantitative colorimetric data that was comparable to that obtained with a conventional plate-reader method. The paper strip was found to be highly stable and could be stored at 4 °C for at least 10 weeks with no loss in performance, as compared to a complete loss in performance within 1 day when the reagents were printed without the stabilizing polysaccharide. The ease of fabrication coupled with the high stability of the printed reagents provides a facile platform for easily manufactured POC sensors.

  3. Functional Biomaterials: Solution Electrospinning and Gelation of Whey Protein and Pullulan

    NASA Astrophysics Data System (ADS)

    Sullivan, Stephanie Tolstedt

    Utilizing biomaterials that are biodegradable, biocompatible and edible serve well for food products as well as biomedical applications. Biomaterials whey protein and pullulan both have these characteristics. Whey proteins (WP) have been used in food products for many years and more recently in pharmaceutical products. They have the ability to form both gels and stable foams. Pullulan (PULL) has also been used in both food and pharmaceutical products, and is a highly water soluble, non-gelling polysaccharide and has been used primarily as a film former. Herein, we investigate the ability of whey protein and pullulan to form nanofibers and gels. Combining their distinct properties allows the ability to uniquely manipulate nanofiber and gel characteristics and behavior for a variety of applications, from food to even tissue scaffolding. First, we determined the electrospinnability of aqueous whey protein solutions. Both whey protein isolate (WPI) and one of its major components beta--lactoglobulin (BLG), either in native or denatured form, yielded interesting micro and nanostructures when electrosprayed; while nanofiber production required blending with a spinnable polymer, poly(ethylene oxide) (PEO). WP:PEO solutions were also successfully electrospun at acidic pH (2≤pH≤3), which could improve shelf life. Fourier Transform Infrared Reflectance (FTIR) analysis of WP:PEO fiber mat indicated some variation in WP secondary structure with varying WPI concentration (as WPI increased, % alpha-helix increased and beta-turn decreased) and pH (as pH decreased from neutral (7.5) to acidic (2), % beta-sheet decreased and alpha-helix increased). X-ray Photoelectron Spectroscopy (XPS) also confirmed the presence of WP on the surface of the blend fibers, augmenting the FTIR analysis. Interestingly, WP:PEO composite nanofibers maintained its fibrous morphology at temperatures as high as 100 °C, above the 60 °C PEO melting point. Further, we show that the blend mats retained a

  4. Genetic Modification of the Marine-Isolated Yeast Aureobasidium melanogenum P16 for Efficient Pullulan Production from Inulin.

    PubMed

    Ma, Zai-Chao; Liu, Nan-Nan; Chi, Zhe; Liu, Guang-Lei; Chi, Zhen-Ming

    2015-08-01

    In this study, in order to directly and efficiently convert inulin into pullulan, the INU1 gene from Kluyveromyces maximum KM was integrated into the genomic DNA and actively expressed in the high pullulan producer Aureobasidium melanogenum P16 isolated from the mangrove ecosystem. After the ability to produce pullulan from inulin by different transformants was examined, it was found that the recombinant strain EI36, one of the transformants, produced 40.92 U/ml of inulinase activity while its wild-type strain P16 only yielded 7.57 U/ml of inulinase activity. Most (99.27 %) of the inulinase produced by the recombinant strain EI36 was secreted into the culture. During the 10-l fermentation, 70.57 ± 1.3 g/l of pullulan in the fermented medium was attained from inulin (138.0 g/l) within 108 h, high inulinase activity (42.03 U/ml) was produced within 60 h, the added inulin was actively hydrolyzed by the secreted inulinase, and most of the reducing sugars were used by the recombinant strain EI36. This confirmed that the genetically engineered yeast of A. melanogenum strain P16 was suitable for direct pullulan production from inulin.

  5. Genetic Modification of the Marine-Isolated Yeast Aureobasidium melanogenum P16 for Efficient Pullulan Production from Inulin.

    PubMed

    Ma, Zai-Chao; Liu, Nan-Nan; Chi, Zhe; Liu, Guang-Lei; Chi, Zhen-Ming

    2015-08-01

    In this study, in order to directly and efficiently convert inulin into pullulan, the INU1 gene from Kluyveromyces maximum KM was integrated into the genomic DNA and actively expressed in the high pullulan producer Aureobasidium melanogenum P16 isolated from the mangrove ecosystem. After the ability to produce pullulan from inulin by different transformants was examined, it was found that the recombinant strain EI36, one of the transformants, produced 40.92 U/ml of inulinase activity while its wild-type strain P16 only yielded 7.57 U/ml of inulinase activity. Most (99.27 %) of the inulinase produced by the recombinant strain EI36 was secreted into the culture. During the 10-l fermentation, 70.57 ± 1.3 g/l of pullulan in the fermented medium was attained from inulin (138.0 g/l) within 108 h, high inulinase activity (42.03 U/ml) was produced within 60 h, the added inulin was actively hydrolyzed by the secreted inulinase, and most of the reducing sugars were used by the recombinant strain EI36. This confirmed that the genetically engineered yeast of A. melanogenum strain P16 was suitable for direct pullulan production from inulin. PMID:25985744

  6. Functional Biomaterials: Solution Electrospinning and Gelation of Whey Protein and Pullulan

    NASA Astrophysics Data System (ADS)

    Sullivan, Stephanie Tolstedt

    Utilizing biomaterials that are biodegradable, biocompatible and edible serve well for food products as well as biomedical applications. Biomaterials whey protein and pullulan both have these characteristics. Whey proteins (WP) have been used in food products for many years and more recently in pharmaceutical products. They have the ability to form both gels and stable foams. Pullulan (PULL) has also been used in both food and pharmaceutical products, and is a highly water soluble, non-gelling polysaccharide and has been used primarily as a film former. Herein, we investigate the ability of whey protein and pullulan to form nanofibers and gels. Combining their distinct properties allows the ability to uniquely manipulate nanofiber and gel characteristics and behavior for a variety of applications, from food to even tissue scaffolding. First, we determined the electrospinnability of aqueous whey protein solutions. Both whey protein isolate (WPI) and one of its major components beta--lactoglobulin (BLG), either in native or denatured form, yielded interesting micro and nanostructures when electrosprayed; while nanofiber production required blending with a spinnable polymer, poly(ethylene oxide) (PEO). WP:PEO solutions were also successfully electrospun at acidic pH (2≤pH≤3), which could improve shelf life. Fourier Transform Infrared Reflectance (FTIR) analysis of WP:PEO fiber mat indicated some variation in WP secondary structure with varying WPI concentration (as WPI increased, % alpha-helix increased and beta-turn decreased) and pH (as pH decreased from neutral (7.5) to acidic (2), % beta-sheet decreased and alpha-helix increased). X-ray Photoelectron Spectroscopy (XPS) also confirmed the presence of WP on the surface of the blend fibers, augmenting the FTIR analysis. Interestingly, WP:PEO composite nanofibers maintained its fibrous morphology at temperatures as high as 100 °C, above the 60 °C PEO melting point. Further, we show that the blend mats retained a

  7. Versatile Types of MRI-Visible Cationic Nanoparticles Involving Pullulan Polysaccharides for Multifunctional Gene Carriers.

    PubMed

    Huang, Yajun; Hu, Hao; Li, Rui-Quan; Yu, Bingran; Xu, Fu-Jian

    2016-02-17

    Owing to the low cytotoxicity and excellent biocompatibility, polysaccharides are good candidates for the development of promising biomaterials. In this paper, a series of magnetic resonance imaging (MRI)-visible cationic polymeric nanoparticles involving liver cell-targeting polysaccharides were flexibly designed for multifunctional gene delivery systems. The pullulan-based vector (PuPGEA) consisting of one liver cell-targeting pullulan backbone and ethanolamine-functionalized poly(glycidyl methacrylate) (denoted by BUCT-PGEA) side chains with abundant hydroxyl units and secondary amine was first prepared by atom transfer radical polymerization. The resultant cationic nanoparticles (PuPGEA-GdL or PuPGEA-GdW) with MRI functions were produced accordingly by assembling PuPGEA with aminophenylboronic acid-modified Gd-DTPA (GdL) or GdW10O36(9-) (GdW) via the corresponding etherification or electrostatic interaction. The properties of the PuPGEA-GdL and PuPGEA-GdW nanoparticles including pDNA condensation ability, cytotoxicity, gene transfection, cellular uptake, and in vitro and in vivo MRI were characterized in details. Such kinds of cationic nanoparticles exhibited good performances in gene transfection in liver cells. PuPGEA-GdW demonstrated much better MRI abilities. The present design of PuPGEA-based cationic nanoparticles with the liver cell-targeting polysaccharides and MRI contrast agents would shed light on the exploration of tumor-targetable multifunctional gene delivery systems. PMID:26841955

  8. Continuous gluconic acid production by isolated yeast-like mould strains of Aureobasidium pullulans.

    PubMed

    Anastassiadis, S; Aivasidis, A; Wandrey, C

    2003-04-01

    By extensive microbial screening, about 50 strains with the ability to secrete gluconic acid were isolated from wild flowers. The strains belong to the yeast-like mould Aureobasidium pullulans (de Bary) Arnaud. In shake flask experiments, gluconic acid concentrations between 23 and 140 g/l were produced within 2 days using a mineral medium. In batch experiments, various important fermentation parameters influencing gluconic acid production by A. pullulans isolate 70 (DSM 7085) were identified. Continuous production of gluconic acid with free-growing cells of the isolated yeast-like microorganisms was studied. About 260 g/l gluconic acid at total glucose conversion could be achieved using continuous stirred tank reactors in defined media with residence times (RT) of about 26 h. The highest space-time-yield of 19.3 g l(-1) x h(-1)) with a gluconic acid concentration of 207.5 g/l was achieved with a RT of 10.8 h. The possibility of gluconic acid production with biomass retention by immobilised cells on porous sinter glass is discussed. The new continuous gluconate fermentation process provides significant advantages over traditional discontinuous operation employing Aspergillus niger. The aim of this work was the development of a continuous fermentation process for the production of gluconic acid. Process control becomes easier, offering constant product quality and quantity.

  9. Superabsorbent polysaccharide hydrogels based on pullulan derivate as antibacterial release wound dressing.

    PubMed

    Li, Huanan; Yang, Jing; Hu, Xiaona; Liang, Jie; Fan, Yujiang; Zhang, Xingdong

    2011-07-01

    To accomplish ideal wound dressing, hydrogels based on a natural polysaccharide, pullulan were synthesized by chemical cross-linking. The tensile strengths of the hydrogel films (1 mm thick) were determined to range from 0.663 to 1.097 MPa in proportion to cross-linking degrees and water contents. The swelling study of the hydrogels in water showed remarkable water absorption property with swelling ratio up to 4000%, which provided the hydrogel with quick hemostatic ability and prevent the wound bed from accumulation of exudates. The water vapor transmission rate and water retention of the hydrogels were found to be in the range of 2213-3498 g/m²/day and 34.74-45.81% (after 6 days), indicating that the hydrogel can maintain a moist environment over wound bed, which could prevent the dehydration of the wound bed and prevent the scab formation. Biocompatibility test revealed that the hydrogels were not cytotoxic. The hydrogel could load antimicrobial agents and effectively suppress bacterial proliferation to protect the wound from bacterial invasion. These results suggest that the pullulan hydrogels prepared in this study may have high potential as new ideal wound-dressing materials.

  10. Natural antifouling compounds produced by a novel fungus Aureobasidium pullulans HN isolated from marine biofilm.

    PubMed

    Gao, Min; Su, Rongguo; Wang, Ke; Li, Xuzhao; Lu, Wei

    2013-12-15

    A fungus, Aureobasidium pullulans, was isolated from marine biofilm and identified. A bioassay-guided fractionation procedure was developed to isolate and purify antifouling compounds from A. pullulans HN. The procedure was: fermentation broth-aeration and addition of sodium thiosulfate-graduated pH and liquid-liquid extraction-SPE purification-GC-MS analysis. Firstly, the fermentation broth was tested for its toxicity. Then it was treated with aeration and addition of sodium thiosulfate, and its toxicity was almost not changed. Lastly, antifouling compounds were extracted at different pH, the extract had high toxicity at pH 2 but almost no toxicity at pH 10, which suggested the toxicants should be fatty acids. The EC50 of the extract against Skeletonema costatum was 90.9 μg ml(-1), and its LC50 against Balanus amphitrete larvae was 22.2 μg ml(-1). After purified by HLB SPE column, the EC50 of the extract against S. costatum was 49.4 μg ml(-1). The myristic and palmitic acids were found as the main toxicants by GC-MS. PMID:24210009

  11. An Injectable Enzymatically Crosslinked Carboxymethylated Pullulan/Chondroitin Sulfate Hydrogel for Cartilage Tissue Engineering.

    PubMed

    Chen, Feng; Yu, Songrui; Liu, Bing; Ni, Yunzhou; Yu, Chunyang; Su, Yue; Zhu, Xinyuan; Yu, Xiaowei; Zhou, Yongfeng; Yan, Deyue

    2016-01-01

    In this study, an enzymatically cross-linked injectable and biodegradable hydrogel system comprising carboxymethyl pullulan-tyramine (CMP-TA) and chondroitin sulfate-tyramine (CS-TA) conjugates was successfully developed under physiological conditions in the presence of both horseradish peroxidase (HRP) and hydrogen peroxide (H2O2) for cartilage tissue engineering (CTTE). The HRP crosslinking method makes this injectable system feasible, minimally invasive and easily translatable for regenerative medicine applications. The physicochemical properties of the mechanically stable hydrogel system can be modulated by varying the weight ratio and concentration of polymer as well as the concentrations of crosslinking reagents. Additionally, the cellular behaviour of porcine auricular chondrocytes encapsulated into CMP-TA/CS-TA hydrogels demonstrates that the hydrogel system has a good cyto-compatibility. Specifically, compared to the CMP-TA hydrogel, these CMP-TA/CS-TA composite hydrogels have enhanced cell proliferation and increased cartilaginous ECM deposition, which significantly facilitate chondrogenesis. Furthermore, histological analysis indicates that the hydrogel system exhibits acceptable tissue compatibility by using a mouse subcutaneous implantation model. Overall, the novel injectable pullulan/chondroitin sulfate composite hydrogels presented here are expected to be useful biomaterial scaffold for regenerating cartilage tissue. PMID:26817622

  12. An Injectable Enzymatically Crosslinked Carboxymethylated Pullulan/Chondroitin Sulfate Hydrogel for Cartilage Tissue Engineering

    NASA Astrophysics Data System (ADS)

    Chen, Feng; Yu, Songrui; Liu, Bing; Ni, Yunzhou; Yu, Chunyang; Su, Yue; Zhu, Xinyuan; Yu, Xiaowei; Zhou, Yongfeng; Yan, Deyue

    2016-01-01

    In this study, an enzymatically cross-linked injectable and biodegradable hydrogel system comprising carboxymethyl pullulan-tyramine (CMP-TA) and chondroitin sulfate-tyramine (CS-TA) conjugates was successfully developed under physiological conditions in the presence of both horseradish peroxidase (HRP) and hydrogen peroxide (H2O2) for cartilage tissue engineering (CTTE). The HRP crosslinking method makes this injectable system feasible, minimally invasive and easily translatable for regenerative medicine applications. The physicochemical properties of the mechanically stable hydrogel system can be modulated by varying the weight ratio and concentration of polymer as well as the concentrations of crosslinking reagents. Additionally, the cellular behaviour of porcine auricular chondrocytes encapsulated into CMP-TA/CS-TA hydrogels demonstrates that the hydrogel system has a good cyto-compatibility. Specifically, compared to the CMP-TA hydrogel, these CMP-TA/CS-TA composite hydrogels have enhanced cell proliferation and increased cartilaginous ECM deposition, which significantly facilitate chondrogenesis. Furthermore, histological analysis indicates that the hydrogel system exhibits acceptable tissue compatibility by using a mouse subcutaneous implantation model. Overall, the novel injectable pullulan/chondroitin sulfate composite hydrogels presented here are expected to be useful biomaterial scaffold for regenerating cartilage tissue.

  13. Isolation and characterization of pullulan-degrading Anoxybacillus species isolated from Malaysian hot springs.

    PubMed

    Chai, Yen Yen; Kahar, Ummirul Mukminin; Md Salleh, Madihah; Md Illias, Rosli; Goh, Kian Mau

    2012-06-01

    Two thermophilic bacteria (SK3-4 and DT3-1) were isolated from the Sungai Klah (SK) and Dusun Tua (DT) hot springs in Malaysia. The cells from both strains were rod-shaped, stained Gram positive and formed endospores. The optimal growth of both strains was observed at 55 degrees C and pH 7. Strain DT3-1 exhibited a higher tolerance to chloramphenicol (100 microg ml(-1)) but showed a lower tolerance to sodium chloride (2%, w/v) compared to strain SK3-4. Phylogenetic analysis based on 16S rRNA gene sequences revealed that both strains belong to the genus Anoxybacillus. High concentrations of 15:0 iso in the fatty acid profiles support the conclusion that both strains belong to the genus Anoxybacillus and exhibit unique fatty acid compositions and percentages compared to other Anoxybacillus species. The DNA G + C contents were 42.0 mol% and 41.8 mol% for strains SK3-4 and DT3-1, respectively. Strains SK3-4 and DT3-1 were able to degrade pullulan and to produce maltotriose and glucose, respectively, as their main end products. Based on phenotypic and chemotaxonomic characteristics, 16S rRNA gene sequences, and the DNA G + C content, we propose that strains SK3-4 and DT3-1 are new pullulan-degrading Anoxybacillus strains.

  14. An Injectable Enzymatically Crosslinked Carboxymethylated Pullulan/Chondroitin Sulfate Hydrogel for Cartilage Tissue Engineering

    PubMed Central

    Chen, Feng; Yu, Songrui; Liu, Bing; Ni, Yunzhou; Yu, Chunyang; Su, Yue; Zhu, Xinyuan; Yu, Xiaowei; Zhou, Yongfeng; Yan, Deyue

    2016-01-01

    In this study, an enzymatically cross-linked injectable and biodegradable hydrogel system comprising carboxymethyl pullulan-tyramine (CMP-TA) and chondroitin sulfate-tyramine (CS-TA) conjugates was successfully developed under physiological conditions in the presence of both horseradish peroxidase (HRP) and hydrogen peroxide (H2O2) for cartilage tissue engineering (CTTE). The HRP crosslinking method makes this injectable system feasible, minimally invasive and easily translatable for regenerative medicine applications. The physicochemical properties of the mechanically stable hydrogel system can be modulated by varying the weight ratio and concentration of polymer as well as the concentrations of crosslinking reagents. Additionally, the cellular behaviour of porcine auricular chondrocytes encapsulated into CMP-TA/CS-TA hydrogels demonstrates that the hydrogel system has a good cyto-compatibility. Specifically, compared to the CMP-TA hydrogel, these CMP-TA/CS-TA composite hydrogels have enhanced cell proliferation and increased cartilaginous ECM deposition, which significantly facilitate chondrogenesis. Furthermore, histological analysis indicates that the hydrogel system exhibits acceptable tissue compatibility by using a mouse subcutaneous implantation model. Overall, the novel injectable pullulan/chondroitin sulfate composite hydrogels presented here are expected to be useful biomaterial scaffold for regenerating cartilage tissue. PMID:26817622

  15. Superabsorbent polysaccharide hydrogels based on pullulan derivate as antibacterial release wound dressing.

    PubMed

    Li, Huanan; Yang, Jing; Hu, Xiaona; Liang, Jie; Fan, Yujiang; Zhang, Xingdong

    2011-07-01

    To accomplish ideal wound dressing, hydrogels based on a natural polysaccharide, pullulan were synthesized by chemical cross-linking. The tensile strengths of the hydrogel films (1 mm thick) were determined to range from 0.663 to 1.097 MPa in proportion to cross-linking degrees and water contents. The swelling study of the hydrogels in water showed remarkable water absorption property with swelling ratio up to 4000%, which provided the hydrogel with quick hemostatic ability and prevent the wound bed from accumulation of exudates. The water vapor transmission rate and water retention of the hydrogels were found to be in the range of 2213-3498 g/m²/day and 34.74-45.81% (after 6 days), indicating that the hydrogel can maintain a moist environment over wound bed, which could prevent the dehydration of the wound bed and prevent the scab formation. Biocompatibility test revealed that the hydrogels were not cytotoxic. The hydrogel could load antimicrobial agents and effectively suppress bacterial proliferation to protect the wound from bacterial invasion. These results suggest that the pullulan hydrogels prepared in this study may have high potential as new ideal wound-dressing materials. PMID:21523902

  16. Cloning, sequencing, and regulation of a xylanase gene from the fungus Aureobasidium pullulans Y-2311-1

    SciTech Connect

    Li, Xin-Liang; Ljungdahl, L.G.

    1994-09-01

    Aureobasidium pullulans Y-2311-1 growing on xylan secretes four major xylanases with different masses and isoelectric points. Two of these enzymes, named APX-I and APX-II, have been purified previously. Their N-terminal amino acid sequences are identical except that APX-I has Asp and APX-II has Asn at position 7. An 83-bp DNA region was amplified by PCR and used as a probe for the xylanase gene cloning. The longest cDNA (xynA) obtained by cDNA cloning and PCR amplification consisted of 895 bp. A. pullulans xynA had an open reading frame encoding a polypeptide of 221 amino acids with a calculated mass of 23,531 Da and contained a putative 34-amino-acid signal peptide in front of the amino terminus of the mature enzyme. Strong homology was found between the deduced amino acid sequence of XynA and some xylanases from bacterial and fungal sources. It is suggested that A. pullulans XynA belongs to the family G glycanases. Northern (RNA blot) analysis revealed that only one transcript of 900 bases was present in cultures grown in medium containing D-xylose or oat spelt xylan. Transcription was completely repressed in the presence of glucose in the medium. Southern blot analysis indicated that A. pullulans xynA was present as a single copy in the genome. Comparison between the genomic and cDNA sequences revealed that one intron of 59 bp was present in the coding region. The data presented suggest that the highly active xylanases, APX-I and APX-II, secreted by A. pullulans are encoded by the same gene. 36 refs., 7 figs., 3 tabs.

  17. Synthesis, characterization, and cytotoxicity of the plasmid EGFP-p53 loaded on pullulan-spermine magnetic nanoparticles

    NASA Astrophysics Data System (ADS)

    Eslaminejad, Touba; Nematollahi-Mahani, Seyed Noureddin; Ansari, Mehdi

    2016-03-01

    Magnetic nanoparticles have been used as effective vehicles for the targeted delivery of therapeutic agents that can be controlled in their concentration and distribution to a desired part of the body by using externally driven magnets. This study focuses on the synthesis, characterization, and functionalization of pullulan-spermine (PS) magnetic nanoparticles for medical applications. Magnetite nanopowder was produced by thermal decomposition of goethite (FeOOH) in oleic acid and 1-octadecene; pullulan-spermine was deposited on the magnetite nanoparticles in the form of pullulan-spermine clusters. EGFP-p53 plasmid was loaded on functionalized iron oleate to transfer into cells. Synthesized nanoparticles were characterized by Fourier transform infrared spectroscopy (FTIR), dynamic light scattering (DLS), vibrating sample magnetometry (VSM), and transmission electron microscopy (TEM). The encapsulation efficiency and drug loading efficiency of the nanocomplexes were tested. FTIR studies showed the presence of oleic acid and 1-octadecene in the iron oleate nanopowder and verified the interaction between spermine and pullulan. The characteristic bands of PS in the spectrum of the pullulan-spermine-coated iron oleate (PSCFO) confirmed that PS covered the surface of the iron oleate particles. TEM studies showed the average size of the iron oleate nanopowder, the PSCFO, and the plasmid-carrying PSCFO (PSCFO/pEGFP-p53) to be 34±12 nm, 100±50 nm and 172±3 nm, respectively. Magnetic measurements revealed that magnetic saturation of the PSCFO was lower in comparison with the iron oleate nanopowder due to the presence of organic compounds in the former. In cytotoxicity tests performed using U87 cells as glioblastoma cells, a 92% survival rate was observed at 50 μg/μl of the plasmid-carrying PSCFO, with an IC50 value of 189 μg/μl.

  18. Modification of functional properties of pullulan-whey protein bionanocomposite films with nanoclay.

    PubMed

    Hassannia-Kolaee, Mahbobeh; Khodaiyan, Faramarz; Shahabi-Ghahfarrokhi, Iman

    2016-02-01

    In this study, biodegradable nanocomposite film composed of pullulan - whey protein isolate (WPI) - montmorillonite (MMT) were developed and characterized as a function of incorporating various amounts of MMT nanoparticles (0, 1, 3 and 5 % wt). Results showed that the water-vapor permeability, moisture content, moisture absorption and water solubility decreased when the nano-MMT content was increased. Tensile strength improved and elongation at break simultaneously decreased with increasing MMT content. The glass transition temperature (Tg(and melting-point temperature (Tm) increased with increasing nano-MMT content. Scanning electron microscope (SEM) and X-ray diffraction (XRD) analysis revealed uniform distribution of MMT into the polymer matrix. Atomic force microscopy (AFM) showed enhancement of films' roughness with increasing MMT content. PMID:27162410

  19. Modification of functional properties of pullulan-whey protein bionanocomposite films with nanoclay.

    PubMed

    Hassannia-Kolaee, Mahbobeh; Khodaiyan, Faramarz; Shahabi-Ghahfarrokhi, Iman

    2016-02-01

    In this study, biodegradable nanocomposite film composed of pullulan - whey protein isolate (WPI) - montmorillonite (MMT) were developed and characterized as a function of incorporating various amounts of MMT nanoparticles (0, 1, 3 and 5 % wt). Results showed that the water-vapor permeability, moisture content, moisture absorption and water solubility decreased when the nano-MMT content was increased. Tensile strength improved and elongation at break simultaneously decreased with increasing MMT content. The glass transition temperature (Tg(and melting-point temperature (Tm) increased with increasing nano-MMT content. Scanning electron microscope (SEM) and X-ray diffraction (XRD) analysis revealed uniform distribution of MMT into the polymer matrix. Atomic force microscopy (AFM) showed enhancement of films' roughness with increasing MMT content.

  20. β-Glucan Derived from Aureobasidium pullulans Is Effective for the Prevention of Influenza in Mice

    PubMed Central

    Muramatsu, Daisuke; Iwai, Atsushi; Aoki, Shiho; Uchiyama, Hirohumi; Kawata, Koji; Nakayama, Yosuke; Nikawa, Yasuhiro; Kusano, Kisato; Okabe, Mitsuyasu; Miyazaki, Tadaaki

    2012-01-01

    β-(1→3)-D-glucans with β-(1→6)-glycosidic linked branches produced by mushrooms, yeast and fungi are known to be an immune activation agent, and are used in anti-cancer drugs or health-promoting foods. In this report, we demonstrate that oral administration of Aureobasidium pullulans-cultured fluid (AP-CF) enriched with the β-(1→3),(1→6)-D-glucan exhibits efficacy to protect mice infected with a lethal titer of the A/Puerto Rico/8/34 (PR8; H1N1) strain of influenza virus. The survival rate of the mice significantly increased by AP-CF administration after sublethal infection of PR8 virus. The virus titer in the mouse lung homogenates was significantly decreased by AP-CF administration. No significant difference in the mRNA expression of inflammatory cytokines, and in the population of lymphocytes was observed in the lungs of mice administered with AP-CF. Interestingly, expression level for the mRNA of virus sensors, RIG-I (retinoic acid-inducible gene-I) and MDA5 (melanoma differentiation-associated protein 5) strongly increased at 5 hours after the stimulation of A. pullulans-produced purified β-(1→3),(1→6)-D-glucan (AP-BG) in murine macrophage-derived RAW264.7 cells. Furthermore, the replication of PR8 virus was significantly repressed by pre-treatment of AP-BG. These findings suggest the increased expression of virus sensors is effective for the prevention of influenza by the inhibition of viral replication with the administration of AP-CF. PMID:22844473

  1. pH-sensitive pullulan-based nanoparticle carrier for adriamycin to overcome drug-resistance of cancer cells.

    PubMed

    Guo, Hua; Liu, Yuanyuan; Wang, Yan; Wu, Jing; Yang, Xiaoying; Li, Rongshan; Wang, Yinsong; Zhang, Ning

    2014-10-13

    Urocanic acid was conjugated to pullulan to synthesize O-urocanyl pullulan (URPA) with degree of substitution (DS) of 8.2%. URPA nanoparticles prepared by dialysis method had spherical shapes and a mean diameter of 156.8 ± 16.8 nm. Adriamycin (ADR) was successfully loaded into URPA nanoparticles and exhibited pH-sensitive in vitro release property. MTT assay showed that ADR-loaded URPA (ADR/URPA) nanoparticles had a significant higher toxicity against drug resistant MCF-7/ADR cells than free ADR, and the reversal index reached up to 9.6. The results of flow cytometry and confocal microscopy showed that URPA nanoparticles efficiently enhanced accumulation and retention of ADR in MCF-7/ADR cells and successfully delivered ADR into cell nucleus. The reversal effect of ADR/URPA nanoparticles on the drug resistance of MCF-7/ADR cells was perhaps related with their cell entry and intracellular drug release mechanisms.

  2. Microfibrillated cellulose and borax as mechanical, O₂-barrier, and surface-modulating agents of pullulan biocomposite coatings on BOPP.

    PubMed

    Cozzolino, Carlo A; Campanella, Gaetano; Türe, Hasan; Olsson, Richard T; Farris, Stefano

    2016-06-01

    Multifunctional composite coatings on bi-oriented polypropylene (BOPP) films were obtained using borax and microfibrillated cellulose (MFC) added to the main pullulan coating polymer. Spectroscopy analyses suggested that a first type of interaction occurred via hydrogen bonding between the C6OH group of pullulan and the hydroxyl groups of boric acid, while monodiol and didiol complexation represented a second mechanism. The deposition of the coatings yielded an increase in the elastic modulus of the entire plastic substrate (from ∼2GPa of the neat BOPP to ∼3.1GPa of the P/B+/MFC-coated BOPP). The addition of MFC yielded a decrease of both static and kinetic coefficients of friction of approximately 22% and 25%, respectively, as compared to the neat BOPP. All composite coatings dramatically increased the oxygen barrier performance of BOPP, especially under dry conditions. The deposition of the high hydrophilic coatings allowed to obtain highly wettable surfaces (water contact angle of ∼18°).

  3. Effect of meadowsweet flower extract-pullulan coatings on rhizopus rot development and postharvest quality of cold-stored red peppers.

    PubMed

    Synowiec, Alicja; Gniewosz, Małgorzata; Kraśniewska, Karolina; Chlebowska-Śmigiel, Anna; Przybył, Jarosław L; Bączek, Katarzyna; Węglarz, Zenon

    2014-01-01

    The study involved an examination of the antifungal activity on red peppers of pullulan coating (P) and pullulan coating containing either water-ethanol (P + eEMF) or ethanol extract of meadowsweet flowers (P + eEMF). Pullulan was obtained from a culture of Aureobasidium pullulans B-1 mutant. Both non-inoculated peppers and those artificially inoculated with Rhizopus arrhizus were coated and incubated at 24 °C for 5 days. The intensity of the decay caused by Rhizopus arrhizus in the peppers with P and P + eEMF coatings was nearly 3-fold lower, and in the case of P + weEMF 5-fold lower, than that observed in the control peppers. Additionally, the P + weEMF coating decreased, almost two-fold the severity of pepper decay compared to other samples. The influence of coating of pepper postharvest quality was examined after 30 days of storage at 6 °C and 70%-75% RH. All coatings formed a thin and well-attached additional layer of an intensified gloss. During storage, color, total soluble solid content and weight loss of coated peppers were subject to lower changes in comparison with uncoated ones. The results indicate the possibility of the application of pullulan coatings containing MFEs as an alternative to the chemical fungicides used to combat pepper postharvest diseases. PMID:25157463

  4. 26 CFR 31.3121(b)(17)-1 - Services in employ of Communist organization.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 15 2010-04-01 2010-04-01 false Services in employ of Communist organization. 31.3121(b)(17)-1 Section 31.3121(b)(17)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF... COLLECTION OF INCOME TAX AT SOURCE Federal Insurance Contributions Act (Chapter 21, Internal Revenue Code...

  5. Process optimization of continuous gluconic acid fermentation by isolated yeast-like strains of Aureobasidium pullulans.

    PubMed

    Anastassiadis, Savas; Aivasidis, Alexander; Wandrey, Christian; Rehm, Hans-Jürgen

    2005-08-20

    This study was focused on the optimization of a new fermentation process for continuous gluconic acid production by the isolated yeast-like strain Aureobasidium pullulans DSM 7085 (isolate 70). Operational fermentation parameters were optimized in chemostat cultures, using a defined glucose medium. Different optima were found for growth and gluconic acid production for each set of operation parameters. Highest productivity was recorded at pH values between 6.5 and 7.0 and temperatures between 29 and 31 degrees C. A gluconic acid concentration higher than 230 g/L was continuously produced at residence times of 12 h. A steady state extracellular gluconic acid concentration of 234 g/L was measured at pH 6.5. 122% air saturation yielded the highest volumetric productivity and product concentration. The biomass-specific productivity increased steadily upon raising air saturation. An intracellular gluconic acid concentration of about 159 g/L (0.83 mol) was determined at 31 degrees C. This is to be compared with an extracellular concentration of 223 g/L (1.16 mol), which indicates the possible existence of an active transport system for gluconic acid secretion, or the presence of extracellular glucose oxidizing enzymes. The new process provides significant advantages over the traditional discontinuous fungi operations. The process control becomes easier, thus offering stable product quality and quantity.

  6. Cholesteryl Pullulan Encapsulated TNF-α Nanoparticles Are an Effective Mucosal Vaccine Adjuvant against Influenza Virus

    PubMed Central

    Nagatomo, Daiki; Taniai, Madoka; Ariyasu, Harumi; Taniguchi, Mutsuko; Aga, Miho; Ariyasu, Toshio; Ohta, Tsunetaka; Fukuda, Shigeharu

    2015-01-01

    We encapsulated tumor necrosis factor-α (TNF-α), a major proinflammatory cytokine, into cholesteryl pullulan (CHP) to prepare TNF/CHP nanoparticles. In this report, we describe the immune-enhancing capability of the nanoparticles to act as a vaccine adjuvant. TNF/CHP nanoparticles showed excellent storage stability and enhanced host immune responses to external immunogens. The nanoparticles were effective via the nasal route of administration for inducing systemic IgG1 as well as mucosal IgA. We applied the nanoparticles in a model experimental influenza virus infection to investigate their adjuvant ability. TNF/CHP nanoparticles combined with a conventional split vaccine protected mice via nasal administration against a lethal challenge of A/PR/8/34 (H1N1) influenza virus. Mechanistic studies showed that the nanoparticles enhanced antigen uptake by dendritic cells (DCs) and moderately induced the expression of inflammation-related genes in nasopharynx lymphoid tissue (NALT), leading to the activation of both B and T cells. Preliminary safety study revealed no severe toxicity to TNF/CHP nanoparticles. Slight-to-moderate influences in nasal mucosa were observed only in the repeated administration and they seemed to be reversible. Our data show that TNF/CHP nanoparticles effectively enhance both humoral and cellular immunity and could be a potential adjuvant for vaccines against infectious diseases, especially in the mucosa. PMID:26421290

  7. Hydrophobicity, thermal and micro-structural properties of whey protein concentrate-pullulan-beeswax films.

    PubMed

    Jafari, Seid Mahdi; Khanzadi, Mehrdad; Mirzaei, Habibollah; Dehnad, Danial; Chegini, Faramarz Khodaian; Maghsoudlou, Yayha

    2015-09-01

    In this research, effects of beeswax (BW) on functional properties of whey protein concentrates (WPC):pullulan (PUL) films were investigated. For this purpose, 0, 10, 20 and 30w/w(glycerol)% BW rates and 30:70, 50:50 and 70:30w/w% WPC:PUL ratios were applied. Films containing 70% WPC:30% PUL (WPC70) and 30% BW (BW30) justified the highest contact angle (92.4°) among all films; SEM micrographs indicated that BW could come toward the surface of films during drying stage and resulted in a higher hydrophobic behavior of bilayer films compared with blend films. WPC70 supplied the lowest T(g) values (36-48 °C) among different proportions of WPC-PUL; the highest melting points were just assured in the absence of BW regardless of combination ratio for WPI:PUL. BW30 films deserved lower roughness rates than BW20 (and even BW10) films, indicating more advantageous microstructure and higher hydrogen connections in BW30 films and justifying similar melting points attained for BW30 films to BW20 or 10 ones. Overall, application of WPC70 and BW30 was recommended to obtain optimum combination of final properties for WPC-PUL-BW bilayer films as SEM exhibited flexible and elastic structures of such films.

  8. Development of ecofriendly bionanocomposite: Whey protein isolate/pullulan films with nano-SiO2.

    PubMed

    Hassannia-Kolaee, Mahbobeh; Khodaiyan, Faramarz; Pourahmad, Rezvan; Shahabi-Ghahfarrokhi, Iman

    2016-05-01

    During the past decade, the limitation of petroleum based polymers, the high price of oil, and the environmental concern were attracted the attention of researchers to develop biobased polymers. The composition of different biopolymers and the reinforcement with nano filler are common methods to improve the drawbacks of biopolymers. In this study whey protein isolate/pullulan (WPI/PUL) films contain 1%, 3%, and 5% (w/w) nano-SiO2 (NS) were prepared by a casting method. Tensile strength of nanocomposite films increased after increasing NS content, but elongation at break decreased, simultaneously. Water absorption, moisture content, solubility in water improved in the wake of increasing NS content because NS increase the cohesiveness of the polymer matrix and improved the barrier and water resistance properties of the films. water vapor permeability of film specimens decreased by increasing NS content. Uniform distribution of NS into polymer matrix was confirmed by scanning electron microscopy (SEM). XRD pattern and thermal analysis revealed increasing crystallinity and increasing Tg of film specimens with increasing NS content, respectively. According to our result WPI/PUL/NS films possess potential to be used as environment friendly packaging films to improve shelf life of food and can be used as promising alternative to petroleum based packaging films.

  9. Cell-laden microengineered pullulan methacrylate hydrogels promote cell proliferation and 3D cluster formation.

    PubMed

    Bae, Hojae; Ahari, Amir F; Shin, Hyeongho; Nichol, Jason W; Hutson, Che B; Masaeli, Mahdokht; Kim, Su-Hwan; Aubin, Hug; Yamanlar, Seda; Khademhosseini, Ali

    2011-01-01

    The ability to encapsulate cells in three-dimensional (3D) environments is potentially of benefit for tissue engineering and regenerative medicine. In this paper, we introduce pullulan methacrylate (PulMA) as a promising hydrogel platform for creating cell-laden microscale tissues. The hydration and mechanical properties of PulMA were demonstrated to be tunable through modulation of the degree of methacrylation and gel concentration. Cells encapsulated in PulMA exhibited excellent viability. Interestingly, while cells did not elongate in PulMA hydrogels, cells proliferated and organized into clusters, the size of which could be controlled by the hydrogel composition. By mixing with gelatin methacrylate (GelMA), the biological properties of PulMA could be enhanced as demonstrated by cells readily attaching to, proliferating, and elongating within the PulMA/GelMA composite hydrogels. These data suggest that PulMA hydrogels could be useful for creating complex, cell-responsive microtissues, especially for applications that require controlled cell clustering and proliferation.

  10. Green synthesis of pullulan stabilized gold nanoparticles for cancer targeted drug delivery

    NASA Astrophysics Data System (ADS)

    Ganeshkumar, Moorthy; Ponrasu, Thangavel; Raja, Modhugoor Devendiran; Subamekala, Muthaiya Kannappan; Suguna, Lonchin

    2014-09-01

    The aim of this study was to synthesize green chemistry based gold nanoparticles using liver specific biopolymer and to develop a liver cancer targeted drug delivery system with enhanced efficacy and minimal side effects. Pullulan stabilized gold nanoparticles (PAuNPs) were coupled with 5-Fluorouracil (5-Fu) and folic acid (Fa) which could be used as a tool for targeted drug delivery and imaging of cancer. The toxicity of 5-Fu, 5-Fu adsorbed gold nanoparticles (5-Fu@AuNPs), Fa-coupled 5-Fu adsorbed gold nanoparticles (5-Fu@AuNPs-Fa), was studied using zebrafish embryo as an in vivo model. The in vitro cytotoxicity of free 5-Fu, 5-Fu@AuNPs, 5-Fu@AuNPs-Fa against HepG2 cells was studied and found that the amount of 5-Fu required to achieve 50% of growth of inhibition (Ic50) was much lower in 5-Fu@AuNP-Fa than in free 5-Fu, 5-Fu@AuNPs. The in vivo biodistribution of PAuNPs showed that higher amount of gold had been accumulated in liver (54.42 ± 5.96 μg) than in other organs.

  11. Discrimination of large maltooligosaccharides from isobaric dextran and pullulan using ion mobility mass spectrometry

    PubMed Central

    Rashid, Abdul M; Saalbach, Gerhard; Bornemann, Stephen

    2013-01-01

    RATIONALE Ion mobility mass spectrometry (IMMS) has previously been shown to resolve small isobaric oligosaccharides, but larger alpha-oligoglucans are also abundant in biology and are of industrial importance. If conformational differences between such isomers are retained in the gas phase, IMMS could be used to address questions in biology and industry. METHODS Negative mode electrospray ionization (ESI) travelling-wave IMMS was used to resolve large isobaric α-glucan ions on the basis of their different gas-phase conformations. α,ω-Dicarboxy-terminated polystyrene was used to calibrate the instrument allowing the collision cross-sections (CCSs) of ions to be determined. RESULTS α-1,4-Linked maltooligosaccharides with a degree of polymerisation of up to 35 could be discriminated from α-1,6-linked dextran and α-1,4/1,6-linked pullulan using IMMS. Fragmentation spectra of ions separated by IMMS could also distinguish isomers. Two conformational isomers of maltohexaose were resolvable by IMMS, likely reflecting extended and V6 helical conformations. IMMS was also able to identify a product within a mixture of maltooligosaccharides treated with the potential anti-tuberculosis drug target Mycobacterium tuberculosis GlgB branching enzyme. CONCLUSIONS Biological samples of complex isobaric oligosaccharides can be analysed using IMMS in the negative mode providing facile analyses and high sensitivity without the need for either derivatisation or chromatographic separation. © 2013 The Authors. Rapid Communications in Mass Spectrometry published by John Wiley & Sons Ltd. PMID:24338967

  12. Development of ecofriendly bionanocomposite: Whey protein isolate/pullulan films with nano-SiO2.

    PubMed

    Hassannia-Kolaee, Mahbobeh; Khodaiyan, Faramarz; Pourahmad, Rezvan; Shahabi-Ghahfarrokhi, Iman

    2016-05-01

    During the past decade, the limitation of petroleum based polymers, the high price of oil, and the environmental concern were attracted the attention of researchers to develop biobased polymers. The composition of different biopolymers and the reinforcement with nano filler are common methods to improve the drawbacks of biopolymers. In this study whey protein isolate/pullulan (WPI/PUL) films contain 1%, 3%, and 5% (w/w) nano-SiO2 (NS) were prepared by a casting method. Tensile strength of nanocomposite films increased after increasing NS content, but elongation at break decreased, simultaneously. Water absorption, moisture content, solubility in water improved in the wake of increasing NS content because NS increase the cohesiveness of the polymer matrix and improved the barrier and water resistance properties of the films. water vapor permeability of film specimens decreased by increasing NS content. Uniform distribution of NS into polymer matrix was confirmed by scanning electron microscopy (SEM). XRD pattern and thermal analysis revealed increasing crystallinity and increasing Tg of film specimens with increasing NS content, respectively. According to our result WPI/PUL/NS films possess potential to be used as environment friendly packaging films to improve shelf life of food and can be used as promising alternative to petroleum based packaging films. PMID:26774376

  13. Disulphide cross linked pullulan based cationic polymer for improved gene delivery and efflux pump inhibition.

    PubMed

    S, Priya S; R, Rekha M

    2016-10-01

    Multidrug resistance is a hurdle to successful cancer chemotherapy. Over expression of P-glycoprotein (P-gp) is a prime contributing factor for drug resistance. In this study, a disulphide cross-linked pullulan-based cationic polymer (PPSS) was synthesized to act simultaneously as gene delivery vehicle and efflux pump inhibitor. The PPSS nanoplexes were of size <200nm with the zeta potential of +15 to +20mV. The cytotoxicity studies using C6 and L929 cells showed that PPSS polymers are non-toxic even at high polymer concentrations. The PPSS/pDNA nanoplex showed superior uptake in confocal microscopy with 97% uptake by flow cytometry. The efficacy of efflux pump inhibition by the PPSS nanoplex was established by the enhanced intracellular retention of DOX. The enhanced cell death by p53/PPSS/DOX nanoplexes was attributed to the synergistic effect of P-gp inhibition and p53 transfection efficiency. Therefore, this multifunctional polymeric system may have significant promise for therapeutic application against cancer drug resistance. PMID:27459414

  14. Occurrence and diversity of marine yeasts in Antarctica environments

    NASA Astrophysics Data System (ADS)

    Zhang, Xue; Hua, Mingxia; Song, Chunli; Chi, Zhenming

    2012-03-01

    A total of 28 yeast strains were obtained from the sea sediment of Antarctica. According to the results of routine identification and molecular characterization, the strains belonged to species of Yarrowia lipolytica, Debaryomyces hansenii, Rhodotorula slooffiae, Rhodotorula mucilaginosa, Sporidiobolus salmonicolor, Aureobasidium pullulans, Mrakia frigida and Guehomyces pullulans, respectively. The Antarctica yeasts have wide potential applications in biotechnology, for some of them can produce β-galactosidase and killer toxins.

  15. Antimicrobial activity of a pullulan-caraway essential oil coating on reduction of food microorganisms and quality in fresh baby carrot.

    PubMed

    Gniewosz, Małgorzata; Kraśniewska, Karolina; Woreta, Marcin; Kosakowska, Olga

    2013-08-01

    This research evaluated the antimicrobial efficacy of pullulan films containing caraway essential oil (CEO). The films were prepared from a 10% of pullulan, containing from 0.12% to 10.0% of CEO. The composition of the CEO was analyzed with the use of gas chromatography. The antimicrobial activity of the CEO was evaluated with the method of serial microdilutions, and the films containing CEO-with the agar diffusion method against selected Gram-negative, Gram-positive bacteria, and fungi. The structure of the film surface and its cross-section were analyzed using a scanning electron microscope (SEM). Analyses were also carried out to determine the efficacy of a pullulan coating with 10% CEO on baby carrots experimentally inoculated with Salmonella enteritidis, Staphylococcus aureus, Saccharomyces cerevisiae, or Aspergillus niger and stored at a room temperature for 7 d. At a concentration of 0.12%, CEO inhibited the growth of all the tested microorganisms. Pullulan films containing 8% to 10% of CEO were active against all tested microorganisms. Populations of S. aureus on carrot samples were reduced by approximately 3 log CFU/g, while those of A. niger and S. cerevisiae by, respectively, 5 and 4 log CFU/g, after 7 d of storage. S. enteritidis was the most resistant among the tested species, since it was not significantly reduced after 7 d of storage. At the end of storage, samples treated with pullulan-caraway oil coating maintained better visual acceptability than control samples. Results of this study suggest the feasibility of applying a pullulan film with incorporated CEO to extend the microbiological stability of minimally processed foods. PMID:23957414

  16. Antimicrobial activity of a pullulan-caraway essential oil coating on reduction of food microorganisms and quality in fresh baby carrot.

    PubMed

    Gniewosz, Małgorzata; Kraśniewska, Karolina; Woreta, Marcin; Kosakowska, Olga

    2013-08-01

    This research evaluated the antimicrobial efficacy of pullulan films containing caraway essential oil (CEO). The films were prepared from a 10% of pullulan, containing from 0.12% to 10.0% of CEO. The composition of the CEO was analyzed with the use of gas chromatography. The antimicrobial activity of the CEO was evaluated with the method of serial microdilutions, and the films containing CEO-with the agar diffusion method against selected Gram-negative, Gram-positive bacteria, and fungi. The structure of the film surface and its cross-section were analyzed using a scanning electron microscope (SEM). Analyses were also carried out to determine the efficacy of a pullulan coating with 10% CEO on baby carrots experimentally inoculated with Salmonella enteritidis, Staphylococcus aureus, Saccharomyces cerevisiae, or Aspergillus niger and stored at a room temperature for 7 d. At a concentration of 0.12%, CEO inhibited the growth of all the tested microorganisms. Pullulan films containing 8% to 10% of CEO were active against all tested microorganisms. Populations of S. aureus on carrot samples were reduced by approximately 3 log CFU/g, while those of A. niger and S. cerevisiae by, respectively, 5 and 4 log CFU/g, after 7 d of storage. S. enteritidis was the most resistant among the tested species, since it was not significantly reduced after 7 d of storage. At the end of storage, samples treated with pullulan-caraway oil coating maintained better visual acceptability than control samples. Results of this study suggest the feasibility of applying a pullulan film with incorporated CEO to extend the microbiological stability of minimally processed foods.

  17. A carboxymethyl cellulase from a marine yeast ( Aureobasidium pullulans 98): Its purification, characterization, gene cloning and carboxymethyl cellulose digestion

    NASA Astrophysics Data System (ADS)

    Rong, Yanjun; Zhang, Liang; Chi, Zhenming; Wang, Xianghong

    2015-10-01

    We have reported that A. pullulans 98 produces a high yield of cellulase. In this study, a carboxymethyl cellulase (CMCase) in the supernatant of the culture of A. pullulans 98 was purified to homogeneity, and the maximum production of CMCase was 4.51 U (mg protein)-1. The SDS-PAGE analysis showed that the molecular mass of the purified CMCase was 67.0 kDa. The optimal temperature of the purified enzyme with considerable thermosensitivity was 40°C, much lower than that of the CMCases from other fungi. The optimal pH of the enzyme was 5.6, and the activity profile was stable in a range of acidity (pH 5.0-6.0). The enzyme was activated by Na+, Mg2+, Ca2+, K+, Fe2+ and Cu2+, however, it was inhibited by Fe3+, Ba2+, Zn2+, Mn2+ and Ag+. K m and V max values of the purified enzyme were 4.7 mg mL-1 and 0.57 µmol L-1 min-1 (mg protein)-1, respectively. Only oligosaccharides with different sizes were released from carboxymethylcellulose (CMC) after hydrolysis with the purified CMCase. The putative gene encoding CMCase was cloned from A. pullulans 98, which contained an open reading frame of 954 bp (EU978473). The protein deduced contained the conserved domain of cellulase superfamily (glucosyl hydrolase family 5). The N-terminal amino acid sequence of the purified CMCase was M-A-P-H-A-E-P-Q-S-Q-T-T-E-Q-T-S-S-G-Q-F, which was consistent with that deduced from the cloned gene. This suggested that the purified CMCase was indeed encoded by the cloned CMCase gene in this yeast.

  18. An indigenous hyperproductive species of Aureobasidium pullulans RYLF-10: influence of fermentation conditions on exopolysaccharide (EPS) production.

    PubMed

    Yadav, Kanchan L; Rahi, Deepak K; Soni, Sanjeev K

    2014-02-01

    In recent years, a significant interest has been generated in discovering and developing exopolysaccharides (EPS) produced by microorganisms, especially fungi due to their multifaceted industrial and pharmacological applications. A number of filamentous and cellular fungi have been explored for this; however, according to the existing literature, the work on exopolysaccharide production by indigenous culture on this aspect is still very less and requires a serious attention. The present work is an attempt in this regard and aims to optimize the submerged culture conditions to produce the exopolysaccharides from an indigenous yeast Aureobasidium pullulans RYLF-10 with respect to several operating parameters in shake flask fermentation. The yeast A. pullulans RYLF-10 was identified by 18s RNA sequencing and detailed study on its nutritional requirements, and environmental conditions for submerged culture have been optimized. The optimal temperature and pH for both the vegetative growth and EPS production were found to be 28 ± 1 °C and 5.0, respectively, while the agitation speed and inoculum size were reported to be 150 rpm and 1 % (v/v), respectively. Sucrose (50 g/l) and yeast extract (1 g/l) were found to be the most suitable carbon and nitrogen sources which worked best in the ratio of 60:1 and resulted in the maximum EPS yield. Similarly, the other variables like growth regulator (riboflavin) and minerals (NaCl + K2HPO4 + MgSO4) altogether resulted in a noteworthy EPS yield of 45.24 g/l which is the maximum yield from this indigenous isolate of A. pullulans RYLF-10. PMID:24293276

  19. Crosslinked pullulan/cellulose acetate fibrous scaffolds for bone tissue engineering.

    PubMed

    Atila, Deniz; Keskin, Dilek; Tezcaner, Ayşen

    2016-12-01

    Natural polymer based fibrous scaffolds have been explored for bone tissue engineering applications; however, their inadequate 3-dimensionality and poor mechanical properties are among the concerns for their use as bone substitutes. In this study, pullulan (P) and cellulose acetate (CA), two polysaccharides, were electrospun at various P/CA ratios (P80/CA20, P50/CA50, and P20/CA80%) to develop 3D fibrous network. The scaffolds were then crosslinked with trisodium trimetaphosphate (STMP) to improve the mechanical properties and to delay fast weight loss. The lowest weight loss was observed for the groups that were crosslinked with P/STMP 2/1 for 10min. Fiber morphologies of P50/CA50 were more uniform without phase separation and this group was crosslinked most efficiently among groups. It was found that mechanical properties of P20/CA80 and P50/CA50 were higher than that of P80/CA20. After crosslinking strain values of P50/CA50 scaffolds were improved and these scaffolds became more stable. Unlike P80/CA20, uncrosslinked P50/CA50 and P20/CA80 were not lost in PBS. Among all groups, crosslinked P50/CA50 scaffolds had more uniform pores; therefore this group was used for bioactivity and cell culture studies. Apatite-like structures were observed on fibers after SBF incubation. Human Osteogenic Sarcoma Cell Line (Saos-2) seeded onto crosslinked P50/CA50 scaffolds adhered and proliferated. The functionality of cells was tested by measuring ALP activity of the cells and the results indicated their osteoblastic differentiation. In vitro tests showed that scaffolds were cytocompatible. To sum up, crosslinked P50/CA50 scaffolds were proposed as candidate cell carriers for bone tissue engineering applications.

  20. Crosslinked pullulan/cellulose acetate fibrous scaffolds for bone tissue engineering.

    PubMed

    Atila, Deniz; Keskin, Dilek; Tezcaner, Ayşen

    2016-12-01

    Natural polymer based fibrous scaffolds have been explored for bone tissue engineering applications; however, their inadequate 3-dimensionality and poor mechanical properties are among the concerns for their use as bone substitutes. In this study, pullulan (P) and cellulose acetate (CA), two polysaccharides, were electrospun at various P/CA ratios (P80/CA20, P50/CA50, and P20/CA80%) to develop 3D fibrous network. The scaffolds were then crosslinked with trisodium trimetaphosphate (STMP) to improve the mechanical properties and to delay fast weight loss. The lowest weight loss was observed for the groups that were crosslinked with P/STMP 2/1 for 10min. Fiber morphologies of P50/CA50 were more uniform without phase separation and this group was crosslinked most efficiently among groups. It was found that mechanical properties of P20/CA80 and P50/CA50 were higher than that of P80/CA20. After crosslinking strain values of P50/CA50 scaffolds were improved and these scaffolds became more stable. Unlike P80/CA20, uncrosslinked P50/CA50 and P20/CA80 were not lost in PBS. Among all groups, crosslinked P50/CA50 scaffolds had more uniform pores; therefore this group was used for bioactivity and cell culture studies. Apatite-like structures were observed on fibers after SBF incubation. Human Osteogenic Sarcoma Cell Line (Saos-2) seeded onto crosslinked P50/CA50 scaffolds adhered and proliferated. The functionality of cells was tested by measuring ALP activity of the cells and the results indicated their osteoblastic differentiation. In vitro tests showed that scaffolds were cytocompatible. To sum up, crosslinked P50/CA50 scaffolds were proposed as candidate cell carriers for bone tissue engineering applications. PMID:27612808

  1. Bone engineering by phosphorylated-pullulan and β-TCP composite.

    PubMed

    Takahata, Tomohiro; Okihara, Takumi; Yoshida, Yasuhiro; Yoshihara, Kumiko; Shiozaki, Yasuyuki; Yoshida, Aki; Yamane, Kentaro; Watanabe, Noriyuki; Yoshimura, Masahide; Nakamura, Mariko; Irie, Masao; Van Meerbeek, Bart; Tanaka, Masato; Ozaki, Toshifumi; Matsukawa, Akihiro

    2015-12-01

    A multifunctional biomaterial with the capacity bond to hard tissues, such as bones and teeth, is a real need for medical and dental applications in tissue engineering and regenerative medicine. Recently, we created phosphorylated-pullulan (PPL), capable of binding to hydroxyapatite in bones and teeth. In the present study, we employed PPL as a novel biocompatible material for bone engineering. First, an in vitro evaluation of the mechanical properties of PPL demonstrated both PPL and PPL/β-TCP composites have higher shear bond strength than materials in current clinical use, including polymethylmethacrylate (PMMA) cement and α-tricalcium phosphate (TCP) cement, Biopex-R. Further, the compressive strength of PPL/β-TCP composite was significantly higher than Biopex-R. Next, in vivo osteoconductivity of PPL/β-TCP composite was investigated in a murine intramedular injection model. Bone formation was observed 5 weeks after injection of PPL/β-TCP composite, which was even more evident at 8 weeks; whereas, no bone formation was detected after injection of PPL alone. We then applied PPL/β-TCP composite to a rabbit ulnar bone defect model and observed bone formation comparable to that induced by Biopex-R. Implantation of PPL/β-TCP composite induced new bone formation at 4 weeks, which was remarkably evident at 8 weeks. In contrast, Biopex-R remained isolated from the surrounding bone at 8 weeks. In a pig vertebral bone defect model, defects treated with PPL/β-TCP composite were almost completely replaced by new bone; whereas, PPL alone failed to induce bone formation. Collectively, our results suggest PPL/β-TCP composite may be useful for bone engineering. PMID:26586655

  2. Microfibrillated cellulose and borax as mechanical, O₂-barrier, and surface-modulating agents of pullulan biocomposite coatings on BOPP.

    PubMed

    Cozzolino, Carlo A; Campanella, Gaetano; Türe, Hasan; Olsson, Richard T; Farris, Stefano

    2016-06-01

    Multifunctional composite coatings on bi-oriented polypropylene (BOPP) films were obtained using borax and microfibrillated cellulose (MFC) added to the main pullulan coating polymer. Spectroscopy analyses suggested that a first type of interaction occurred via hydrogen bonding between the C6OH group of pullulan and the hydroxyl groups of boric acid, while monodiol and didiol complexation represented a second mechanism. The deposition of the coatings yielded an increase in the elastic modulus of the entire plastic substrate (from ∼2GPa of the neat BOPP to ∼3.1GPa of the P/B+/MFC-coated BOPP). The addition of MFC yielded a decrease of both static and kinetic coefficients of friction of approximately 22% and 25%, respectively, as compared to the neat BOPP. All composite coatings dramatically increased the oxygen barrier performance of BOPP, especially under dry conditions. The deposition of the high hydrophilic coatings allowed to obtain highly wettable surfaces (water contact angle of ∼18°). PMID:27083358

  3. Edible Active Coatings Based on Pectin, Pullulan, and Chitosan Increase Quality and Shelf Life of Strawberries (Fragaria ananassa).

    PubMed

    Treviño-Garza, Mayra Z; García, Santos; del Socorro Flores-González, Ma; Arévalo-Niño, Katiushka

    2015-08-01

    Edible active coatings (EACs) based on pectin, pullulan, and chitosan incorporated with sodium benzoate and potassium sorbate were employed to improve the quality and shelf life of strawberries. Fruits were washed, disinfected, coated by dipping, packed, and stored at 4 °C for 15 d. Application of EACs reduced (P < 0.05) weight loss and fruit softening and delayed alteration of color (redness) and total soluble solids content. In contrast, pH and titratable acidity were not affected (P > 0.05) throughout storage, and ascorbic acid content was maintained in pectin-EAC coated strawberries. Microbiological analyses showed that application of EACs reduced (P < 0.05) microbial growth (total aerobic counts, molds, and yeasts) on strawberries. Chitosan-EAC coated strawberries presented the best results in microbial growth assays. Sensory quality (color, flavor, texture, and acceptance) improved and decay rate decreased (P < 0.05) in pectin-EAC, pullulan-EAC, and chitosan-EAC coated strawberries. In conclusion, EACs based on polysaccharides improved the physicochemical, microbiological, and sensory characteristics, increasing the shelf life of strawberries from 6 (control) to 15 d (coated fruits). PMID:26189365

  4. Edible Active Coatings Based on Pectin, Pullulan, and Chitosan Increase Quality and Shelf Life of Strawberries (Fragaria ananassa).

    PubMed

    Treviño-Garza, Mayra Z; García, Santos; del Socorro Flores-González, Ma; Arévalo-Niño, Katiushka

    2015-08-01

    Edible active coatings (EACs) based on pectin, pullulan, and chitosan incorporated with sodium benzoate and potassium sorbate were employed to improve the quality and shelf life of strawberries. Fruits were washed, disinfected, coated by dipping, packed, and stored at 4 °C for 15 d. Application of EACs reduced (P < 0.05) weight loss and fruit softening and delayed alteration of color (redness) and total soluble solids content. In contrast, pH and titratable acidity were not affected (P > 0.05) throughout storage, and ascorbic acid content was maintained in pectin-EAC coated strawberries. Microbiological analyses showed that application of EACs reduced (P < 0.05) microbial growth (total aerobic counts, molds, and yeasts) on strawberries. Chitosan-EAC coated strawberries presented the best results in microbial growth assays. Sensory quality (color, flavor, texture, and acceptance) improved and decay rate decreased (P < 0.05) in pectin-EAC, pullulan-EAC, and chitosan-EAC coated strawberries. In conclusion, EACs based on polysaccharides improved the physicochemical, microbiological, and sensory characteristics, increasing the shelf life of strawberries from 6 (control) to 15 d (coated fruits).

  5. Blending of soluble corn fiber with pullulan, sorbitol, or fructose attenuates glycemic and insulinemic responses in the dog and affects hydrolytic digestion in vitro.

    PubMed

    de Godoy, M R C; Knapp, B K; Bauer, L L; Swanson, K S; Fahey, G C

    2013-08-01

    The objective of these experiments was to measure in vitro hydrolytic digestion and glycemic and insulinemic responses of select carbohydrate blends, all containing the novel carbohydrate soluble corn fiber (SCF). Two SCF that varied in their method of production were used to formulate the carbohydrate blends. One set of blends contained a SCF that was spray dried (SCFsd) and then blended with different amounts of either pullulan, sorbitol, or fructose. The other set of blends contained a SCF produced using longer evaporation time (SCF) and then blended with different ratios of pullulan, sorbitol, and fructose. Free sugar concentrations found in the individual SCFsd and SCF substrates were low but varied. Spray-dried soluble corn fiber had a reduced free sugar concentration compared with SCF (2.8 vs. 14.2%). Glucose was the main free sugar found in both SCFsd and SCF but at different concentrations (2.7 vs. 12.7%, respectively). The majority of the SCFsd blends were completely hydrolyzed to their monosaccharide components. Glucose accounted for most of the hydrolyzed monosaccharides for SCFsd and all the SCFsd blends. Hydrolyzed monosaccharide concentrations for the SCF:pullulan:sorbitol:fructose blends followed similar trends to the SCFsd blends where greater percentages of fructose and sorbitol resulted in decreased (P < 0.05) hydrolyzed monosaccharide concentrations. The SCFsd blends had intermediate to high amounts of monosaccharides released as a result of in vitro hydrolytic digestion. The SCFsd:pullulan blends were more digestible in vitro (approximately 91%; P < 0.05) than SCFsd:fructose or SCFsd:sorbitol. Total released monosaccharides were high in SCFsd blends containing either 50% fructose or sorbitol, but the combination resulted in reduced concentrations of glucose released (P < 0.05). The SCF:pullulan:sorbitol:fructose blends also had intermediate to high released monosaccharides as a result of in vitro hydrolytic digestion. All SCF blends resulted in

  6. 26 CFR 17.1 - Industrial development bonds used to provide solid waste disposal facilities; temporary rules.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... solid waste disposal facilities; temporary rules. 17.1 Section 17.1 Internal Revenue INTERNAL REVENUE... UNDER 26 U.S.C. 103(c) § 17.1 Industrial development bonds used to provide solid waste disposal... substantially all the proceeds of which are used to provide solid waste disposal facilities. Section...

  7. 26 CFR 17.1 - Industrial development bonds used to provide solid waste disposal facilities; temporary rules.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... solid waste disposal facilities; temporary rules. 17.1 Section 17.1 Internal Revenue INTERNAL REVENUE... UNDER 26 U.S.C. 103(c) § 17.1 Industrial development bonds used to provide solid waste disposal... substantially all the proceeds of which are used to provide solid waste disposal facilities. Section...

  8. Production of L-sorbitol from L-fructose by Aureobasidium pullulans LP23 isolated from soy sauce mash.

    PubMed

    Sasahara, Hiroyuki; Izumori, Ken

    2005-08-01

    A strain LP23 that can convert L-fructose to L-sorbitol was isolated from soy sauce mash and identified as Aureobasidium pullulans. The cells grown on L-arabinose were found to have relatively high L-fructose to L-sorbitol conversion potential. Addition of erythritol to the reaction mixture considerably accelerated the conversion rate of L-fructose to L-sorbitol. During the conversion reaction, erythritol was added to the reaction mixture at 8-h intervals to maintain the concentration of erythritol at 1.0%. The final conversion ratios were 82.8%, 95.3%, 92.4%, and 42.6% using washed cells when the concentrations of L-fructose were 1.0%, 2.0%, 5.0% and 10.0%, respectively. The product from L-fructose was identified as L-sorbitol by HPLC analysis, infrared spectroscopy, optical rotation and melting point measurements.

  9. Cyanoethylation of the glucans dextran and pullulan: Substitution pattern and formation of nanostructures and entrapment of magnetic nanoparticles

    PubMed Central

    Fiege, Kathrin; Lünsdorf, Heinrich; Atarijabarzadeh, Sevil

    2012-01-01

    Summary Cyanoethylglucans with a degree of substitution in the range of 0.74 to 2.40 for dextran and 0.84 to 2.42 for pullulan were obtained by Michael addition of acrylonitrile to the glucans under various conditions. Products were thoroughly characterized, comprising elementary analysis, NMR and ATR–IR spectroscopy, and analysis of the substituent distribution in the glucosyl units by GC–FID and GC–MS of the constituting monosaccharide derivatives. Nanostructuring of the highly substituted cyanoethylpolysaccharides was performed by dialysis against a non-solvent. In the presence of ferromagnetic iron-oxide nanoparticles, multicore cyanoethylglucan-coated ferromagnetic nanoparticles were formed by selective entrapment. The specific interaction between cyano groups and iron could be proven. The size distribution and morphology of the nanoparticles were analyzed by dynamic light scattering (DLS), scanning electron microscopy (SEM) and energy-filtered transmission electron microscopy (EF–TEM) with parallel electron energy loss spectroscopy (PEELS). PMID:22563354

  10. Adaptation and Transcriptome Analysis of Aureobasidium pullulans in Corncob Hydrolysate for Increased Inhibitor Tolerance to Malic Acid Production

    PubMed Central

    Zou, Xiang; Wang, Yongkang; Tu, Guangwei; Zan, Zhanquan; Wu, Xiaoyan

    2015-01-01

    Malic acid is a dicarboxylic acid widely used in the food industry, and is also a potential C4 platform chemical. Corncob is a low-cost renewable feedstock from agricultural industry. However, side-reaction products (furfural, 5-hydroxymethylfurfural (HMF), formic acid, and acetic acid) that severely hinder fermentation are formed during corncob pretreatment. The process for producing malic acid from a hydrolysate of corncob was investigated with a polymalic acid (PMA)-producing Aureobasidium pullulans strain. Under the optimal hydrolysate sugar concentration 110 g/L, A. pullulans was further adapted in an aerobic fibrous bed bioreactor (AFBB) by gradually increasing the sugar concentration of hydrolysate. After nine batches of fermentation, the production and productivity of malic acid reached 38.6 g/L and 0.4 g/L h, respectively, which was higher than that in the first batch (27.6 g/L and 0.29 g/L h, respectively). The adapted strain could grow under the stress of 0.5 g/L furfural, 3 g/L HMF, 2g/L acetic acid, and 0.5 g/L formic acid, whereas the wild type did not. Transcriptome analysis revealed that the differentially expressed genes were related to carbohydrate transport and metabolism, lipid transport and metabolism, signal transduction mechanism, redox metabolism, and energy production and conversion under 0.5 g/L furfural and 3 g/L HMF stress conditions. In total, 42 genes in the adapted strain were upregulated by 15-fold or more, and qRT-PCR also confirmed that the expression levels of key genes (i.e. SIR, GSS, CYS, and GSR) involved in sulfur assimilation pathway were upregulated by over 10-fold in adapted strain for cellular protection against oxidative stress. PMID:25793624

  11. Variations in grain lipophilic phytochemicals, proteins and resistance to Fusarium spp. growth during grain storage as affected by biological plant protection with Aureobasidium pullulans (de Bary).

    PubMed

    Wachowska, Urszula; Tańska, Małgorzata; Konopka, Iwona

    2016-06-16

    Modern agriculture relies on an integrated approach, where chemical treatment is reduced to a minimum and replaced by biological control that involves the use of active microorganisms. The effect of the antagonistic yeast-like fungus Aureobasidium pullulans on proteins and bioactive compounds (alkylresorcinols, sterols, tocols and carotenoids) in winter wheat grain and on the colonization of wheat kernels by fungal microbiota, mainly Fusarium spp. pathogens, was investigated. Biological treatment contributed to a slight increase contents of tocols, alkylresorcinols and sterols in grain. At the same time, the variation of wheat grain proteins was low and not significant. Application of A. pullulans enhanced the natural yeast colonization after six months of grain storage and inhibited growth of F. culmorum pathogens penetrating wheat kernel. This study demonstrated that an integrated approach of wheat grain protection with the use of the yeast-like fungus A. pullulans reduced kernel colonization by Fusarium spp. pathogens and increased the content of nutritionally beneficial phytochemicals in wheat grain without a loss of gluten proteins responsible for baking value. PMID:27055191

  12. Induction of stilbene phytoalexins in grapevine (Vitis vinifera) and transgenic stilbene synthase-apple plants (Malus domestica) by a culture filtrate of Aureobasidium pullulans.

    PubMed

    Rühmann, Susanne; Pfeiffer, Judith; Brunner, Philipp; Szankowski, Iris; Fischer, Thilo C; Forkmann, Gert; Treutter, Dieter

    2013-11-01

    Products containing the epiphytic yeast Aureobasidium pullulans are commercially available and applied by fruit growers to prevent several fungal and bacterial diseases of fruit trees. The proposed beneficial mechanisms relate to limitations of space and nutrients for the pathogens in presence of the rapidly proliferating yeast cells. These explanations ignore the potential of yeasts to elicit the plant's defense. Our experiments aim at clarifying if an autoclaved and centrifuged suspension of A. pullulans may induce defense mechanisms. As a model system, the biosynthesis and accumulation of stilbene phytoalexins in callus and shoots of grapevine Vitis vinifera grown in vitro was used. Yeast application to the plant tissue stimulated stilbene biosynthesis, sometimes at the cost of flavonoids. The expression of the gene encoding stilbene synthase was enhanced and the enzyme showed higher activity while chalcone synthase activity and expression was reduced in some cases. An accumulation of stilbenes was also found in transgenic apple trees (Malus domestica cv. Holsteiner Cox) harboring the stilbene synthase-gene under control of its own promoter. These results clearly show that the application of A. pullulans may induce defense mechanisms of the treated plants.

  13. 26 CFR 31.3401(a)(17)-1 - Remuneration for services performed on a boat engaged in catching fish.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... engaged in catching fish. 31.3401(a)(17)-1 Section 31.3401(a)(17)-1 Internal Revenue INTERNAL REVENUE... Remuneration for services performed on a boat engaged in catching fish. (a) Remuneration for services performed on or after December 31, 1954, by an individual on a boat engaged in catching fish or other forms...

  14. 26 CFR 31.3401(a)(17)-1 - Remuneration for services performed on a boat engaged in catching fish.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... engaged in catching fish. 31.3401(a)(17)-1 Section 31.3401(a)(17)-1 Internal Revenue INTERNAL REVENUE... Remuneration for services performed on a boat engaged in catching fish. (a) Remuneration for services performed on or after December 31, 1954, by an individual on a boat engaged in catching fish or other forms...

  15. 26 CFR 31.3401(a)(17)-1 - Remuneration for services performed on a boat engaged in catching fish.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... engaged in catching fish. 31.3401(a)(17)-1 Section 31.3401(a)(17)-1 Internal Revenue INTERNAL REVENUE... Remuneration for services performed on a boat engaged in catching fish. (a) Remuneration for services performed on or after December 31, 1954, by an individual on a boat engaged in catching fish or other forms...

  16. 26 CFR 31.3401(a)(17)-1 - Remuneration for services performed on a boat engaged in catching fish.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... engaged in catching fish. 31.3401(a)(17)-1 Section 31.3401(a)(17)-1 Internal Revenue INTERNAL REVENUE... Remuneration for services performed on a boat engaged in catching fish. (a) Remuneration for services performed on or after December 31, 1954, by an individual on a boat engaged in catching fish or other forms...

  17. 26 CFR 31.3401(a)(17)-1 - Remuneration for services performed on a boat engaged in catching fish.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... engaged in catching fish. 31.3401(a)(17)-1 Section 31.3401(a)(17)-1 Internal Revenue INTERNAL REVENUE... Remuneration for services performed on a boat engaged in catching fish. (a) Remuneration for services performed on or after December 31, 1954, by an individual on a boat engaged in catching fish or other forms...

  18. Biocontrol activity of an alkaline serine protease from Aureobasidium pullulans expressed in Pichia pastoris against four postharvest pathogens on apple.

    PubMed

    Banani, Houda; Spadaro, Davide; Zhang, Dianpeng; Matic, Slavica; Garibaldi, Angelo; Gullino, Maria Lodovica

    2014-07-16

    The yeast-like fungus Aureobasidium pullulans PL5 is a microbial antagonist against postharvest pathogens of fruits. The strain is able to produce hydrolases, including glucanases, chitinases and proteases. The alkaline serine protease gene ALP5 from A. pullulans was cloned, inserted into the vector pPIC9 to construct pPIC9/ALP5, and then expressed in Pichia pastoris strain KM71. ALP5 had a molecular mass of 42.9kDa after 5days growth with 1% methanol induction at 28°C. The recombinant protease expressed in P. pastoris showed its highest activity under alkaline conditions (at pH10) and a temperature of 50°C. The antifungal activity of the recombinant protease was investigated against Penicillium expansum, Botrytis cinerea, Monilinia fructicola and Alternaria alternata in vitro and on apple. The recombinant protease reduced significantly the spore germination and the germ tube length of the tested pathogens in PDB medium. The highest level of protease efficacy was observed against M. fructicola and B. cinerea, whereas a lower efficacy was observed against P. expansum and A. alternata indicating a possible effect of the pathogen cell wall composition on the proteolytic activity of the recombinant protease. The presence of protease was able to cause the swelling of the hyphae of B. cinerea, under an optical microscope. The recombinant protease expressed in P. pastoris was more active against the pathogens in vitro than the same enzyme expressed in E. coli in previous studies. The efficacy of ALP5 was also evaluated against the pathogens in vivo on cv Golden Delicious apples. The protease was more efficient in controlling M. fructicola, B. cinerea and P. expansum than A. alternata. However, the extent of the activity was dependent on the enzyme concentration and the length of fruit storage. This study demonstrated the capacity of the alkaline serine protease to keep its enzymatic activity for some days in the unfavorable environment of the fruit wounds. The alkaline

  19. Intensification of β-poly(L: -malic acid) production by Aureobasidium pullulans ipe-1 in the late exponential growth phase.

    PubMed

    Cao, Weifeng; Luo, Jianquan; Zhao, Juan; Qiao, Changsheng; Ding, Luhui; Qi, Benkun; Su, Yi; Wan, Yinhua

    2012-07-01

    β-Poly(malic acid) (PMLA) has attracted industrial interest because this polyester can be used as a prodrug or for drug delivery systems. In PMLA production by Aureobasidium pullulans ipe-1, it was found that PLMA production was associated with cell growth in the early exponential growth phase and dissociated from cell growth in the late exponential growth phase. To enhance PMLA production in the late phase, different fermentation modes and strategies for controlling culture redox potential (CRP) were studied. The results showed that high concentrations of produced PMLA (above 40 g/l) not only inhibited PMLA production, but also was detrimental to cell growth. Moreover, when CRP increased from 57 to 100 mV in the late exponential growth phase, the lack of reducing power in the broth also decreased PMLA productivity. PMLA productivity could be enhanced by repeated-batch culture to maintain cell growth in the exponential growth phase, or by cell-recycle culture with membrane to remove the produced PMLA, or by maintaining CRP below 70 mV no matter which kind of fermentation mode was adopted. Repeated-batch culture afforded a high PMLA concentration (up to 63.2 g/l) with a productivity of 1.15 g l(-1) h(-1). Cell-recycle culture also confirmed that PMLA production by the strain ipe-1 was associated with cell growth.

  20. Continuous gluconic acid production by the yeast-like Aureobasidium pullulans in a cascading operation of two bioreactors.

    PubMed

    Anastassiadis, Savas; Rehm, Hans-Jürgen

    2006-12-01

    The application of a new developed process for the continuous production of gluconic acid using a cascade of two bioreactors in a continuous process is shown reaching the highest concentration of gluconic acid described in the literature for continuous culture fermentation. Very high gluconic acid concentrations of 272-308 g/l have been achieved under continuous cultivation of free-growing cells of Aureobasidium pullulans in the first bioreactor at residence times (RT) between 19.5 and 24 h with formation rates for the generic product between 12.7 and 13.9 g/(l h). Gluconic acid, 350-370 g/l, was continuously reached in the second bioreactor at a total RT of 30.8-37 h with R (j) of 9.2-12 g/(l h). The highest specific gluconic acid production (m (p)) of 3.6 g/(g h) was found in the first bioreactor at the lowest RT of 19.5 h. The highest selectivity of 93.6% was determined in the first bioreactor as well. Complete glucose consumption was obtained at 37 h total residence time in the second bioreactor. Gluconic acid, 433 g/l, was continuously produced in the second bioreactor at a total RT of 37 h.

  1. Polycan, a β-glucan from Aureobasidium pullulans SM-2001, mitigates ovariectomy-induced osteoporosis in rats

    PubMed Central

    Jung, Mi Young; Kim, Joo Wan; Kim, Ki Young; Choi, Seong Hun; Ku, Sae Kwang

    2016-01-01

    The present study aimed to investigate the protective effects of Polycan, a β-glucan from Aureobasidium pullulans SM-2001, in a rat model of ovariectomy-induced osteoporosis. Ovariectomized (OVX) rats were orally administered 31.25, 62.5 or 125 mg/kg/day Polycan for 126 days, and alterations in body weight, bone mineral content, bone mineral density, failure load, histological profiles and histomorphometric indices were analyzed. In particular, serum levels of osteocalcin, bone-specific alkaline phosphatase (bALP), calcium and phosphorus, and the urine deoxypyridinoline/creatinine ratio, were measured. Furthermore, the femur, tibia and lumbar vertebrae were harvested from all rats, and histomorphometrical analyses were conducted in order to assess the mass and structure of the bones, and the rates of bone resorption and formation. One group of rats was treated with alendronate, which served as the reference drug. The results of the present study suggested that Polycan treatment was able to inhibit ovariectomy-induced alterations in bone resorption and turnover in a dose-dependent manner. In addition, the serum expression levels of bALP and all histomorphometrical indices for bone formation were markedly increased in the Polycan-treated groups. These results indicated that Polycan was able to preserve bone mass and strength, and increase the rate of bone formation in OVX rats; thus suggesting that Polycan may be considered a potential effective anti-osteoporosis agent. PMID:27588046

  2. A Novel Biosurfactant Produced by Aureobasidium pullulans L3-GPY from a Tiger Lily Wild Flower, Lilium lancifolium Thunb.

    PubMed Central

    Kim, Jong Shik; Lee, In Kyoung; Yun, Bong Sik

    2015-01-01

    Yeast biosurfactants are important biotechnological products in the food industry, and they have medical and cosmeceutical applications owing to their specific modes of action, low toxicity, and applicability. Thus, we have isolated and examined biosurfactant-producing yeast for various industrial and medical applications. A rapid and simple method was developed to screen biosurfactant-producing yeasts for high production of eco-friendly biosurfactants. Using this method, several potential niches of biosurfactant-producing yeasts, such as wild flowers, were investigated. We successfully selected a yeast strain, L3-GPY, with potent surfactant activity from a tiger lily, Lilium lancifolium Thunb. Here, we report the first identification of strain L3-GPY as the black yeast Aureobasidium pullulans. In addition, we isolated a new low-surface-tension chemical, designated glycerol-liamocin, from the culture supernatant of strain L3-GPY through consecutive chromatography steps, involving an ODS column, solvent partition, silica gel, Sephadex LH-20, and an ODS Sep-Pak cartridge column. The chemical structure of glycerol-liamocin, determined by mass spectrometry and nuclear magnetic resonance spectroscopy, indicates that it is a novel compound with the molecular formula C33H62O12. Furthermore, glycerol-liamocin exhibited potent biosurfactant activity (31 mN/m). These results suggest that glycerol-liamocin is a potential novel biosurfactantfor use in various industrial applications. PMID:25849549

  3. Resistant starch and pullulan reduce postprandial glucose, insulin, and GLP-1, but have no effect on satiety in healthy humans.

    PubMed

    Klosterbuer, Abby S; Thomas, William; Slavin, Joanne L

    2012-12-01

    The aim of this study was to determine the effects of three novel fibers on satiety and serum parameters. In a randomized, double-blind, crossover design, fasted subjects (n=20) consumed a low-fiber control breakfast or one of four breakfasts containing 25 g of fiber from soluble corn fiber (SCF) or resistant starch (RS), alone or in combination with pullulan (SCF+P and RS+P). Visual analog scales assessed appetite, and blood samples were collected to measure glucose, insulin, ghrelin, and glucagon-like peptide-1 (GLP-1). The fiber treatments did not influence satiety or energy intake compared to control. RS+P significantly reduced glucose, insulin, and GLP-1, but neither SCF treatment differed from control. To conclude, these fibers have little impact on satiety when provided as a mixed meal matched for calories and macronutrients. Additional research regarding the physiological effects of these novel fibers is needed to guide their use as functional ingredients in food products.

  4. Experimental high gradient testing of a 17.1 GHz photonic band-gap accelerator structure

    NASA Astrophysics Data System (ADS)

    Munroe, Brian J.; Zhang, JieXi; Xu, Haoran; Shapiro, Michael A.; Temkin, Richard J.

    2016-03-01

    We report the design, fabrication, and high gradient testing of a 17.1 GHz photonic band-gap (PBG) accelerator structure. Photonic band-gap (PBG) structures are promising candidates for electron accelerators capable of high-gradient operation because they have the inherent damping of high order modes required to avoid beam breakup instabilities. The 17.1 GHz PBG structure tested was a single cell structure composed of a triangular array of round copper rods of radius 1.45 mm spaced by 8.05 mm. The test assembly consisted of the test PBG cell located between conventional (pillbox) input and output cells, with input power of up to 4 MW from a klystron supplied via a TM01 mode launcher. Breakdown at high gradient was observed by diagnostics including reflected power, downstream and upstream current monitors and visible light emission. The testing procedure was first benchmarked with a conventional disc-loaded waveguide structure, which reached a gradient of 87 MV /m at a breakdown probability of 1.19 ×10-1 per pulse per meter. The PBG structure was tested with 100 ns pulses at gradient levels of less than 90 MV /m in order to limit the surface temperature rise to 120 K. The PBG structure reached up to 89 MV /m at a breakdown probability of 1.09 ×10-1 per pulse per meter. These test results show that a PBG structure can simultaneously operate at high gradients and low breakdown probability, while also providing wakefield damping.

  5. Cloning, characterization, expression and antifungal activity of an alkaline serine protease of Aureobasidium pullulans PL5 involved in the biological control of postharvest pathogens.

    PubMed

    Zhang, Dianpeng; Spadaro, Davide; Valente, Silvia; Garibaldi, Angelo; Gullino, Maria Lodovica

    2012-02-15

    An alkaline protease gene was amplified from genomic DNA and cDNA of the antagonistic yeast-like fungus Aureobasidium pullulans PL5, a biocontrol agent effective against Monilinia laxa on stone fruit and Botrytis cinerea and Penicillium expansum on pome fruits. An open reading frame of 1248 bp encoding a 415-amino acid (aa) protein with a calculated molecular weight (M(r)) of 42.9 kDa and an isoelectric point (pI) of 4.5 was characterized. The cDNAALP5 gene had an 18-amino acid signal peptide, one N-gylcosylation, one histidine active site, and one serine active site. The ALP5 gene with a M(r) of 1351 bp contained two introns. One intron was of 54 bp, while the other was of 50 bp. Protein BLAST and phylogenetic tree analysis of the deduced amino sequences from the cDNAALP5 gene showed that the encoded protein had 100% homology to a protease enzyme (ALP2) of a sea strain of A. pullulans, suggesting that the protein ALP5 was an alkaline serine protease. Expression of ALP5 in Escherichia coli BL21 (DE3), followed by identification with Western-blotting, purification with Ni-NTA and analysis of enzymatic activity, yielded an homogeneous recombinant ALP5 which hydrolysed the substrate casein and inhibited the mycelial growth of the pathogens. At its optimal pH of 10.0 and reaction temperature of 50°C, the recombinant protease exhibited the highest activity towards the substrate casein, though the highest stability was at lower temperatures and pH between 7.0 and 9.0. This study provided the direct evidence that extracellular proteases secreted by the antagonist A. pullulans PL5 played a role in the biocontrol activities against some postharvest pathogens of apple and peach.

  6. Inhibition of UVB-induced skin damage by exopolymers from Aureobasidium pullulans SM-2001 in hairless mice.

    PubMed

    Kim, Kyung Hu; Park, Soo Jin; Lee, Young Joon; Lee, Ji Eun; Song, Chang Hyun; Choi, Seong Hun; Ku, Sae Kwang; Kang, Su Jin

    2015-02-01

    Because antioxidants from natural sources may be an effective approach to the treatment and prevention of UV radiation-induced skin damage, the effects of purified exopolymers from Aureobasidium pullulans SM-2001 ('E-AP-SM2001') were evaluated in UVB-induced hairless mice. E-AP-SM2001 consists of 1.7% β-1,3/1,6-glucan, fibrous polysaccharides and other organic materials, such as amino acids, and mono- and di-unsaturated fatty acids (linoleic and linolenic acids) and shows anti-osteoporotic and immunomodulatory effects, through antioxidant and anti-inflammatory mechanisms. Hairless mice were treated topically with vehicle, E-AP-SM2001 stock and two and four times diluted solutions once per day for 15 weeks against UVB irradiation (three times per week at 0.18 J/cm(2) ). The following parameters were evaluated in skin samples: myeloperoxidase (MPO) activity, cytokine levels [interleukin (IL)-1β and IL-10], endogenous antioxidant content (glutathione, GSH), malondialdehyde (MDA) levels, superoxide anion production; matrix metalloproteases (MMP-1, -9 and -13), GSH reductase and Nox2 (gp91phox) mRNA levels, and immunoreactivity for nitrotyrosine (NT), 4-hydroxynonenal (HNE), caspase-3, and cleaved poly(ADP-ribose) polymerase (PARP). Photoageing was induced by UVB irradiation through ROS-mediated inflammation, which was related to the depletion of endogenous antioxidants, activation of MMPs and keratinocyte apoptosis. Topical treatment with all three doses of E-AP-SM2001 and 5 nm myricetin attenuated the UV-induced depletion of GSH, activation of MMPs, production of IL-1β, the decrease in IL-10 and keratinocyte apoptosis. In this study, E-AP-SM2001 showed potent inhibitory effects against UVB-induced skin photoageing. Thus, E-AP-SM2001 may be useful as a functional ingredient in cosmetics, especially as a protective agent against UVB-induced skin photoageing.

  7. Pullulan/dextran/nHA macroporous composite beads for bone repair in a femoral condyle defect in rats.

    PubMed

    Schlaubitz, Silke; Derkaoui, Sidi Mohammed; Marosa, Lydia; Miraux, Sylvain; Renard, Martine; Catros, Sylvain; Le Visage, Catherine; Letourneur, Didier; Amédée, Joëlle; Fricain, Jean-Christophe

    2014-01-01

    The repair of bone defects is of particular interest for orthopedic, oral, maxillofacial, and dental surgery. Bone loss requiring reconstruction is conventionally addressed through bone grafting. Depending on the size and the location of the defect, this method has limits and risks. Biomaterials can offer an alternative and have features supporting bone repair. Here, we propose to evaluate the cellular penetration and bone formation of new macroporous beads based on pullulan/dextran that has been supplemented with nanocrystalline hydroxyapatite in a rat model. Cross-linked beads of 300-500 µm diameters were used in a lateral femoral condyle defect and analyzed by magnetic resonance imaging, micro-computed tomography, and histology in comparison to the empty defects 15, 30, and 70 days after implantation. Inflammation was absent for both conditions. For empty defects, cellularisation and mineralization started from the periphery of the defect. For the defects containing beads, cellular structures filling out the spaces between the scaffolds with increasing interconnectivity and trabecular-like organization were observed over time. The analysis of calcified sections showed increased mineralization over time for both conditions, but was more pronounced for the samples containing beads. Bone Mineral Density and Bone Mineral Content were both significantly higher at day 70 for the beads in comparison to empty defects as well as compared with earlier time points. Analysis of newly formed tissue around the beads showed an increase of osteoid tissue, measured as percentage of the defect surface. This study suggests that the use of beads for the repair of small size defects in bone may be expanded on to meet the clinical need for a ready-to-use fill-up material that can favor bone formation and mineralization, as well as promote vessel ingrowth into the defect site.

  8. Pullulan/dextran/nHA Macroporous Composite Beads for Bone Repair in a Femoral Condyle Defect in Rats

    PubMed Central

    Schlaubitz, Silke; Derkaoui, Sidi Mohammed; Marosa, Lydia; Miraux, Sylvain; Renard, Martine; Catros, Sylvain; Le Visage, Catherine; Letourneur, Didier; Amédée, Joëlle; Fricain, Jean-Christophe

    2014-01-01

    The repair of bone defects is of particular interest for orthopedic, oral, maxillofacial, and dental surgery. Bone loss requiring reconstruction is conventionally addressed through bone grafting. Depending on the size and the location of the defect, this method has limits and risks. Biomaterials can offer an alternative and have features supporting bone repair. Here, we propose to evaluate the cellular penetration and bone formation of new macroporous beads based on pullulan/dextran that has been supplemented with nanocrystalline hydroxyapatite in a rat model. Cross-linked beads of 300–500 µm diameters were used in a lateral femoral condyle defect and analyzed by magnetic resonance imaging, micro-computed tomography, and histology in comparison to the empty defects 15, 30, and 70 days after implantation. Inflammation was absent for both conditions. For empty defects, cellularisation and mineralization started from the periphery of the defect. For the defects containing beads, cellular structures filling out the spaces between the scaffolds with increasing interconnectivity and trabecular-like organization were observed over time. The analysis of calcified sections showed increased mineralization over time for both conditions, but was more pronounced for the samples containing beads. Bone Mineral Density and Bone Mineral Content were both significantly higher at day 70 for the beads in comparison to empty defects as well as compared with earlier time points. Analysis of newly formed tissue around the beads showed an increase of osteoid tissue, measured as percentage of the defect surface. This study suggests that the use of beads for the repair of small size defects in bone may be expanded on to meet the clinical need for a ready-to-use fill-up material that can favor bone formation and mineralization, as well as promote vessel ingrowth into the defect site. PMID:25330002

  9. The $17.1 billion problem: the annual cost of measurable medical errors.

    PubMed

    Van Den Bos, Jill; Rustagi, Karan; Gray, Travis; Halford, Michael; Ziemkiewicz, Eva; Shreve, Jonathan

    2011-04-01

    At a minimum, high-quality health care is care that does not harm patients, particularly through medical errors. The first step in reducing the large number of harmful medical errors that occur today is to analyze them. We used an actuarial approach to measure the frequency and costs of measurable US medical errors, identified through medical claims data. This method focuses on the analysis of comparative rates of illness, using mathematical models to assess the risk of occurrence and to project costs to the total population. We estimate that the annual cost of measurable medical errors that harm patients was $17.1 billion in 2008. Pressure ulcers were the most common measurable medical error, followed by postoperative infections and by postlaminectomy syndrome, a condition characterized by persistent pain following back surgery. A total of ten types of errors account for more than two-thirds of the total cost of errors, and these errors should be the first targets of prevention efforts.

  10. Incorporation of nisin Z and lauric arginate into pullulan films to inhibit foodborne pathogens associated with fresh and ready-to-eat muscle foods.

    PubMed

    Pattanayaiying, Rinrada; H-Kittikun, Aran; Cutter, Catherine N

    2015-08-17

    A combination of food grade compounds with edible films, used to inhibit foodborne pathogens associated with fresh or further processed muscle foods, is receiving considerable attention. In this study, pullulan films containing lauric arginate (LAE) and nisin Z (produced by Lactococcus lactis subsp. lactis I8-7-3 and isolated from catfish gut), alone or in combination, were investigated for controlling foodborne pathogens on fresh and further processed muscle foods after long-term refrigerated storage. Salmonella Typhimurium and Salmonella Enteritidis on raw turkey breast slices wrapped with a film containing LAE or the combination of LAE with nisin Z were reduced throughout the experiment, 2.5 to 4.5 log10 CFU/cm(2) and 3.5 to 5.1 log10 CFU/cm(2), respectively. Film containing a combination of LAE with nisin Z reduced Staphylococcus aureus and Listeria monocytogenes Scott A inoculated onto ham surfaces by approximately 5.53 and 5.62 log10 CFU/cm(2), respectively during refrigerated storage. Escherichia coli O157:H7, O111, and O26 also were reduced by >4 log 10CFU/cm(2) on raw beef slices after treatment with the combination film and refrigerated storage. The results obtained from this study indicate the LAE- and LAE-nisin Z-containing pullulan films displayed excellent inhibition against foodborne pathogens on fresh and further processed muscle foods.

  11. Incorporation of nisin Z and lauric arginate into pullulan films to inhibit foodborne pathogens associated with fresh and ready-to-eat muscle foods.

    PubMed

    Pattanayaiying, Rinrada; H-Kittikun, Aran; Cutter, Catherine N

    2015-08-17

    A combination of food grade compounds with edible films, used to inhibit foodborne pathogens associated with fresh or further processed muscle foods, is receiving considerable attention. In this study, pullulan films containing lauric arginate (LAE) and nisin Z (produced by Lactococcus lactis subsp. lactis I8-7-3 and isolated from catfish gut), alone or in combination, were investigated for controlling foodborne pathogens on fresh and further processed muscle foods after long-term refrigerated storage. Salmonella Typhimurium and Salmonella Enteritidis on raw turkey breast slices wrapped with a film containing LAE or the combination of LAE with nisin Z were reduced throughout the experiment, 2.5 to 4.5 log10 CFU/cm(2) and 3.5 to 5.1 log10 CFU/cm(2), respectively. Film containing a combination of LAE with nisin Z reduced Staphylococcus aureus and Listeria monocytogenes Scott A inoculated onto ham surfaces by approximately 5.53 and 5.62 log10 CFU/cm(2), respectively during refrigerated storage. Escherichia coli O157:H7, O111, and O26 also were reduced by >4 log 10CFU/cm(2) on raw beef slices after treatment with the combination film and refrigerated storage. The results obtained from this study indicate the LAE- and LAE-nisin Z-containing pullulan films displayed excellent inhibition against foodborne pathogens on fresh and further processed muscle foods. PMID:26001063

  12. Role of a GATA-type transcriptional repressor Sre1 in regulation of siderophore biosynthesis in the marine-derived Aureobasidium pullulans HN6.2.

    PubMed

    Chi, Zhe; Wang, Xing-Xing; Geng, Qian; Chi, Zhen-Ming

    2013-12-01

    The GATA-type transcriptional repressor structural gene SRE1 was isolated from both the genomic DNA and mRNA of the marine yeast Aureobasidium pullulans HN6.2 by inverse PCR and RACE. An open reading frame (ORF) of 1,002 bp encoding a 334 amino acid protein (a calculated isoelectric point: 8.6) with a calculated molecular weight of 35.1 kDa was characterized. The corresponding gene had one single intron of 51 bp, and in its promoter two putative 5'-HGATAR-3' sequences could be recognized. The deduced protein from the cloned gene contained two conserved zinc-finger domains [Cys-(X2)-Cys-(X17)-Cys-(X2)-Cys)], nine sequences of Ser(Thr)-Pro-X-X which was characteristics of the regulator, and one cysteine-rich central domain which was located between the two zinc fingers. The SRE1 gene in A. pullulans HN6.2 was disrupted by integrating the hygromycin B phosphotransferase gene into the ORF of the SRE1 gene using homologous recombination. Two hundreds of the disruptants (Δsre1) (one of them was named R6) obtained still synthesized both intracellular and extracellular siderophores in the presence of added Fe(3+) and the expression of the SidA gene encoding L-ornithine N(5)-oxygenase in the disruptant R6 was also partially derepressed in the presence of added Fe(3+). The colonies of the disruptant R6 grown on the iron-replete medium with 1.5 and 2.0 mM Fe(3+) and also with 1.5 mM Fe(2+) became brown. In contrast, A. pullulans HN6.2 could not grow in the iron-replete medium with 1.5 mM and 2.0 mM Fe(3+). The brown-colored colonies of the disruptant R6 also had high level of siderophore and iron.

  13. Efficient Delivery of DOX to Nuclei of Hepatic Carcinoma Cells in the Subcutaneous Tumor Model Using pH-Sensitive Pullulan-DOX Conjugates.

    PubMed

    Li, Huanan; Cui, Yani; Sui, Junhui; Bian, Shaoquan; Sun, Yong; Liang, Jie; Fan, Yujiang; Zhang, Xingdong

    2015-07-29

    A series of pullulan-doxorubicin conjugates (Pu-DOXs) were investigated for effectively delivering DOX to nuclei of hepatic carcinoma cells in subcutaneous tumor model. These Pu-DOXs were prepared by conjugating DOX onto pullulan molecule via pH-responsive hydrazone bond using spacers with different alkane chain length. The highest drug loading content of Pu-DOXs went up to nearly 50%, and the diameter of Pu-DOX nanoparticles ranged from 50 to 170 nm, as measured by DLS and TEM. These Pu-DOX nanoparticles could rapidly release DOX in the acidic environment at pH = 5.0 while being kept relatively stable in neural conditions. The in vitro cell coculture experiments revealed that these Pu-DOX nanoparticles were selectively internalized by hepatic carcinoma cells through receptor-mediated endocytosis via asialoglycoprotein receptor on the hepatic carcinoma cell surface. DOX was rapidly released from Pu-DOX nanoparticles in acidic endosome/lysosome, diffused into cell nuclei due to its strong affinity to nucleic acid, inhibited the cell proliferation, and accelerated the cell apoptosis. In the nude mice subcutaneous hepatic carcinoma model, Pu-DOX nanoparticles efficiently accumulated in the tumor site through the enhanced permeation and retention effect. Then DOX was specifically internalized by hepatic carcinoma cells and rapidly diffused into the nuclei of cells. Compared with the control group in in vivo experiments, these Pu-DOX nanoparticles effectively inhibited solid tumor growth, prolonging the lifetime of the experimental animal. These pH sensitive nanoparticles might provide an important clinical implication for targeted hepatic carcinoma therapy with high efficiency and low systematic toxicity.

  14. Biodistribution of charged 17.1A photoimmunoconjugates in a murine model of hepatic metastasis of colorectal cancer

    PubMed Central

    Hamblin, M R; Governatore, M Del; Rizvi, I; Hasan, T

    2000-01-01

    Optimizing photodynamic therapy involves attempting to increase both the absolute tumour content of photosensitizer and the selectivity between tumour and surrounding normal tissue. One reason why photodynamic therapy has not been considered suitable for treatment of metastatic tumours in the liver, is the poor selectivity of conventional photosensitizers for tumour compared to normal liver. This report details an alternative approach to increasing this selectivity by the use of antibody-targeted photosensitizers (or photoimmunoconjugates) to target intrahepatic tumours caused by human colorectal cancer cells in the nude mouse, and explores the role of molecular charge on the tumour-targeting efficiency of macromolecules. The murine monoclonal antibody 17.1A (which recognizes an antigen expressed on HT 29 cells) was used to prepare site-specific photoimmunoconjugates with the photosensitizer chlorine6. The conjugates had either a predominant cationic or anionic charge and were injected i.v. into tumour-bearing mice. Biodistribution 3 or 24 h later was measured by extraction of tissue samples and quantitation of chlorine6 content by fluorescence spectroscopy. The photoimmunoconjugates were compared to the polylysine conjugates in an attempt to define the effect of molecular charge as well as antibody targeting. The anionic 17.1A conjugate delivered more than twice as much photosensitizer to the tumour at 3 h than other species (5 times more than the cationic 17.1A conjugate) and had a tumour:normal liver ratio of 2.5. Tumour-to-liver ratios were greater than one for most compounds at 3 h but declined at 24 h. Tumour-to-skin ratios were high (> 38) for all conjugates but not for free chlorine6. Cationic species had a high uptake in the lungs compared to anionic species. The photoimmunoconjugates show an advantage over literature reports of other photosensitizers, which can result in tumour:normal liver ratios of less than 1. © 2000 Cancer Research Campaign http

  15. Effect of whey protein isolate-pullulan edible coatings on the quality and shelf life of freshly roasted and freeze-dried Chinese chestnut.

    PubMed

    Gounga, M E; Xu, S-Y; Wang, Z; Yang, W G

    2008-05-01

    Harvested chestnut is characterized by a short shelf life, exposing many Chinese producers to a storage problem as product losses are very high. The objective of this study was to develop a suitable technology to extend the shelf life of harvested chestnut fruits for commercial use. The effect of whey protein isolate-pullulan (WPI-Pul) coating on fresh-roasted chestnuts (FRC) and roasted freeze-dried chestnut (RFDC) quality and shelf life was studied under 2 different storage temperature (4 and 20 degrees C) conditions. Coatings were formed directly onto the surface of the fruits by dipping them into a film solution. SEM micrographs showed homogeneous WPI-Pul to cover the whole surface of chestnut with good adherence and perfect integrity. Moisture loss or gain, fruit quality, and shelf life were evaluated by weight loss or gain, surface color development, and visible decay during the storage period of 15 to 120 d at 4 and 20 degrees C, respectively. WPI-Pul coating had a low, yet significant effect on reducing moisture loss and decay incidence of FRC, hence delaying changes in their external color. The results were satisfactory when the coating was done with freeze-drying at low temperature storage, thus improving the quality and increasing the shelf life. This provides an alternative strategy to minimize the significant losses in harvested chestnut.

  16. Oral administration of the Aureobasidium pullulans-derived β-glucan effectively prevents the development of high fat diet-induced fatty liver in mice

    PubMed Central

    Aoki, Shiho; Iwai, Atsushi; Kawata, Koji; Muramatsu, Daisuke; Uchiyama, Hirofumi; Okabe, Mitsuyasu; Ikesue, Masahiro; Maeda, Naoyoshi; Uede, Toshimitsu

    2015-01-01

    Aureobasidium pullulans-derived β-glucan (AP-PG) consisting of a β-(1,3)-linked glucose main chain and β-(1,6)-linked glucose branches is taken as a supplement to improve health. This study demonstrates that oral administration of AP-PG is effective to prevent the development of high-fat diet (HFD)-induced fatty liver in mice. Here, C57BL/6N mice were fed with a normal diet or HFD, and AP-PG diluted in drinking water was administered orally. After 16 weeks, the serological analysis showed that HFD-induced high blood cholesterol and triglyceride levels were reduced by the oral administration of AP-PG. Further, HFD induced-fatty liver was significantly reduced by the oral administration of AP-PG. The triglyceride accumulation in the liver was also significantly reduced in mice administered AP-PG. Liver injury as indicated by an increase in serum alanine aminotransferase (ALT) in the HFD-fed mice was significantly reduced in the mice administered AP-PG orally, and the gene expression of cholesterol 7 alpha-hydroxylase (CYP7A1) which is known to be involved in cholesterol degradation in the liver was significantly increased in the AP-PG administered mice. These results suggest the possibility that the oral administration of AP-PG is effective to prevent the development of non-alcoholic fatty liver disease (NAFLD). PMID:26179949

  17. Stimulation with the Aureobasidium pullulans-produced β-glucan effectively induces interferon stimulated genes in macrophage-like cell lines

    PubMed Central

    Muramatsu, Daisuke; Kawata, Koji; Aoki, Shiho; Uchiyama, Hirofumi; Okabe, Mitsuyasu; Miyazaki, Tadaaki; Kida, Hiroshi; Iwai, Atsushi

    2014-01-01

    A β-(1,3),(1,6)-D-glucan produced by A. pullulans (AP-PG) is known to be an immune stimulating agent. In this study, we demonstrate that the stimulation with AP-PG effectively induces the interferon (IFN) stimulated genes (ISGs) in macrophage-like cell lines. The ISGs, Mx1, ISG15, and viperin mRNAs were significantly increased in RAW264.7 cells after stimulation with AP-PG. The stimulation with AP-PG transiently induced IFN-β mRNA. However, the expression of viperin mRNA was also increased after stimulation with AP-PG even when new protein synthesis was completely blocked by treatment with cycloheximide. Further, in IFN-α receptor knockdown RAW264.7 cells, AP-PG stimulation more effectively induced viperin mRNA compared with that of IFN-α stimulation. The phosphorylation of Ser 727 in STAT1 involved in the enhancement of STAT1 activation was immediately increased after stimulation with AP-PG. In addition, viperin mRNA expression induced after stimulation with IFN-α was significantly increased by combined stimulation with AP-PG. These results suggest that stimulation with AP-PG effectively induces the ISGs through the induction of IFN and the enhancement of STAT1-mediated transcriptional activation. PMID:24759061

  18. Novel hydrogels based on carboxyl pullulan and collagen crosslinking with 1, 4-butanediol diglycidylether for use as a dermal filler: initial in vitro and in vivo investigations.

    PubMed

    Li, Xian; Xue, Wenjiao; Zhu, Chenhui; Fan, Daidi; Liu, Yannan; XiaoxuanMa

    2015-12-01

    Novel hydrogels based on carboxyl pullulan (PC) and human-like collagen (HLC) crosslinking with 1,4-butanediol diglycidyl ether (BDDE) are promising soft fillers for tissue engineering due to their highly tunable properties. Recent studies, however, have shown that incorporating hyaluronic acid and BDDE results in hydrogels with a microporous structure, a large pore size and high porosity, which reduce cell adhesion and enhance degradation in vivo. To improve biocompatibility and prevent biodegradation, the use of PC to replace hyaluronic acid in the fabrication of PC/BDDE (PCB) and PC/BDDE/HLC (PCBH) hydrogels was investigated. Preparation of gels with PC is a promising strategy due to the high reactivity, superb selectivity, and mild reaction conditions of PC. In particular, the Schiff base reaction of HLC and PC produces the novel functional group -RCONHR' in PCBH hydrogels. Twenty-four weeks after subcutaneous injection of either PCB or PCBH hydrogel in mice, the surrounding tissue inflammation, enzymatic response and cell attachment were better compared to hyaluronic acid-based hydrogels. However, the biocompatibility, cytocompatibility and non-biodegradability of PCBH were milder than those of the PCB hydrogels both in vivo and in vitro. These results show that the proposed use of PC and HLC for the fabrication of hydrogels is a promising strategy for generating soft filler for tissue engineering.

  19. Novel hydrogels based on carboxyl pullulan and collagen crosslinking with 1, 4-butanediol diglycidylether for use as a dermal filler: initial in vitro and in vivo investigations.

    PubMed

    Li, Xian; Xue, Wenjiao; Zhu, Chenhui; Fan, Daidi; Liu, Yannan; XiaoxuanMa

    2015-12-01

    Novel hydrogels based on carboxyl pullulan (PC) and human-like collagen (HLC) crosslinking with 1,4-butanediol diglycidyl ether (BDDE) are promising soft fillers for tissue engineering due to their highly tunable properties. Recent studies, however, have shown that incorporating hyaluronic acid and BDDE results in hydrogels with a microporous structure, a large pore size and high porosity, which reduce cell adhesion and enhance degradation in vivo. To improve biocompatibility and prevent biodegradation, the use of PC to replace hyaluronic acid in the fabrication of PC/BDDE (PCB) and PC/BDDE/HLC (PCBH) hydrogels was investigated. Preparation of gels with PC is a promising strategy due to the high reactivity, superb selectivity, and mild reaction conditions of PC. In particular, the Schiff base reaction of HLC and PC produces the novel functional group -RCONHR' in PCBH hydrogels. Twenty-four weeks after subcutaneous injection of either PCB or PCBH hydrogel in mice, the surrounding tissue inflammation, enzymatic response and cell attachment were better compared to hyaluronic acid-based hydrogels. However, the biocompatibility, cytocompatibility and non-biodegradability of PCBH were milder than those of the PCB hydrogels both in vivo and in vitro. These results show that the proposed use of PC and HLC for the fabrication of hydrogels is a promising strategy for generating soft filler for tissue engineering. PMID:26354254

  20. The mitochondrial protein import component, TRANSLOCASE OF THE INNER MEMBRANE17-1, plays a role in defining the timing of germination in Arabidopsis.

    PubMed

    Wang, Yan; Law, Simon R; Ivanova, Aneta; van Aken, Olivier; Kubiszewski-Jakubiak, Szymon; Uggalla, Vindya; van der Merwe, Margaretha; Duncan, Owen; Narsai, Reena; Whelan, James; Murcha, Monika W

    2014-11-01

    In Arabidopsis (Arabidopsis thaliana), small gene families encode multiple isoforms for many of the components of the mitochondrial protein import apparatus. There are three isoforms of the TRANSLOCASE OF THE INNER MEMBRANE17 (Tim17). Transcriptome analysis indicates that AtTim17-1 is only detectable in dry seed. In this study, two independent transfer DNA insertional mutant lines of tim17-1 exhibited a germination-specific phenotype, showing a significant increase in the rate of germination. Microarray analyses revealed that Attim17-1 displayed alterations in the temporal sequence of transcriptomic events during germination, peaking earlier compared with the wild type. Promoter analysis of AtTim17-1 further identified an abscisic acid (ABA)-responsive element, which binds ABA-responsive transcription factors, acting to repress the expression of AtTim17-1. Attim17-1 dry seeds contained significantly increased levels of ABA and gibberellin, 2- and 5-fold, respectively. These results support the model that mitochondrial biogenesis is regulated in a tight temporal sequence of events during germination and that altering mitochondrial biogenesis feeds back to alter the germination rate, as evidenced by the altered levels of the master regulatory hormones that define germination.

  1. 17.1%-Efficient Multi-Scale-Textured Black Silicon Solar Cells without Dielectric Antireflection Coating: Preprint

    SciTech Connect

    Toor, F.; Page, M. R.; Branz, H. M.; Yuan, H. C.

    2011-07-01

    In this work we present 17.1%-efficient p-type single crystal Si solar cells with a multi-scale-textured surface and no dielectric antireflection coating. Multi-scale texturing is achieved by a gold-nanoparticle-assisted nanoporous etch after conventional micron scale KOH-based pyramid texturing (pyramid black etching). By incorporating geometric enhancement of antireflection, this multi-scale texturing reduces the nanoporosity depth required to make silicon 'black' compared to nanoporous planar surfaces. As a result, it improves short-wavelength spectral response (blue response), previously one of the major limiting factors in 'black-Si' solar cells. With multi-scale texturing, the spectrum-weighted average reflectance from 350- to 1000-nm wavelength is below 2% with a 100-nm deep nanoporous layer. In comparison, roughly 250-nm deep nanopores are needed to achieve similar reflectance on planar surface. Here, we characterize surface morphology, reflectivity and solar cell performance of the multi-scale textured solar cells.

  2. Modulating the Growth and Imatinib Sensitivity of Chronic Myeloid Leukemia Stem/Progenitor Cells with Pullulan/MicroRNA Nanoparticles In Vitro.

    PubMed

    Ma, Wenjuan; Liu, Jian; Xie, Jundan; Zhang, Xiuyan; Zhou, Haixia; Yao, Hong; Zhang, Weiqi; Guo, Dawei; Zhu, Lingying; Xiao, Lun; Wu, Depei; Xu, Haiyan; Chen, Suning; Zhao, Yun

    2015-11-01

    Chronic myeloid leukemia (CML) originates from normal hematopoietic stem cells acquiring Philadelphia chromosome (Ph) to generate BCR-ABL fusion gene whose protein product has deregulated tyrosine kinase activity. Specific inhibitors against BCR-ABL, such as Imatinib mesylate (IM), have greatly improved CML management; however, no single agent is a cure yet. Delivery of microRNA (miRNA) using non-viral vectors has been utilized to inhibit various cancer cells; however, the efficacy of this approach to target CML stem/progenitor cells has not been elucidated. In this study, we firstly validated that spermine-introduced pullulan (Ps) was a robust non-viral vector for delivery of miRNA to CML cells, including the CD34+ cells from clinical isolates. We then found that the miR-181a/RALA (V-ral simian leukemia viral oncogene homolog A) axis was aberrantly expressed in the CML CD34+ cells. The delivery of miR-181a specifically inhibited the growth of CML CD34+ cells, possibly via the inhibition of RALA. In contrast, miR-181a did not evidently affect the normal hematopoietic CD34+ cells. In addition, miR-181a increased IM sensitivity of the CD34+ CML cells. Taken together, we have therefore demonstrated that the delivery of miR-181a using Ps to CML stem/progenitor cells leads to their growth inhibition and enhancement of IM sensitivity, which will possibly be beneficial to CML treatment. PMID:26554155

  3. Stimulation of Macrophages with the β-Glucan Produced by Aureobasidium pullulans Promotes the Secretion of Tumor Necrosis Factor-Related Apoptosis Inducing Ligand (TRAIL)

    PubMed Central

    Kawata, Koji; Iwai, Atsushi; Muramatsu, Daisuke; Aoki, Shiho; Uchiyama, Hirofumi; Okabe, Mitsuyasu; Hayakawa, Sumio; Takaoka, Akinori; Miyazaki, Tadaaki

    2015-01-01

    A β-glucan produced by Aureobasidium pullulans (AP-PG) is consisting of a β-(1,3)-linked main chain with β-(1,6)-linked glucose side residues. Various β-glucans consisting of β-(1,3)-linked main chain including AP-PG are believed to exhibit anti-tumor activities, and actually, anti-tumor activities of AP-PG in mice have been demonstrated. In this study, we demonstrate that stimulation with AP-PG induces TRAIL expression in mouse and human macrophage-like cell lines. TRAIL is known to be a cytokine which specifically induces apoptosis in transformed cells, but not in untransformed cells. The expression of TRAIL mRNA after stimulation with AP-PG was increased in RAW264.7 cells, Mono Mac 6 cells, and macrophage-differentiated THP-1 cells. The mRNA expression of TNF-α and FasL is only weakly increased after stimulation with AP-PG. The induction activity of TRAIL by curdlan, a bacterial β-glucan, was very similar to that by AP-PG in RAW264.7 cells, but weaker in macrophage-differentiated THP-1 cells. Activation of caspases was found in HeLa cells after treatment with the supernatant of cultured medium from AP-PG-stimulated Mono Mac 6 cells, and was inhibited by the anti-TRAIL neutralizing antibody. These findings suggest that the stimulation with AP-PG effectively induces TRAIL in macrophages, and that it may be related to apoptosis induction of tumor cells. PMID:25875639

  4. Effects of co-fermented Pleurotus eryngii stalk residues and soybean hulls by Aureobasidium pullulans on performance and intestinal morphology in broiler chickens.

    PubMed

    Lai, L P; Lee, M T; Chen, C S; Yu, B; Lee, T T

    2015-12-01

    Soybean hulls are a by-product of soybean processing for oil and meal production; Pleurotus eryngii stalk residues (PESR) are by-products of the edible portion of the fruiting body enriched in bioactive metabolites. This study evaluated the effects of co-fermented PESR and soybean hulls with Aureobasidium pullulans on performance and intestinal morphology in broiler chickens. The in vitro experimental results showed that xylananse and mannanase activity of solid-state fermented soybean hulls (100% SBH) and soybean hulls partially replaced with PESR (75:25, SHP) reached peak at day 12; solid-state fermentation (SSF) enhanced the total phenolic content and trolox equivalency in both products as well. Additionally, FSHP had higher xylotriose and mannobiose levels than fermented FSBH did. A total of 400 broilers (Ross 308) were assigned randomly into four groups receiving the basal diet (control) or the basal diet supplemented with 0.5% fermented SBH (0.5% FSBH), 0.5% fermented SBHP (0.5% FSHP) and 1.0% fermented SBHP (1.0% FSHP) until 35 d of age, respectively. Results demonstrated that 0.5% FSHP addition increased body weight gain as compared with corresponding normal diet fed control in birds during entire experimental period. Compared with the control group, 0.5% FSHP group significantly increased the ratio of lactic acid bacteria to Clostridium perfringens in ceca as well as ileum villus height and jejunum villus height/crypt depth ratio of 35 d old birds. In conclusion, 0.5% FSHP supplementation in the diet could obtain not only improved body weight gain, but optimal intestinal morphology by exerting its bioactive metabolite properties when fed to broilers. PMID:26467005

  5. Effects of co-fermented Pleurotus eryngii stalk residues and soybean hulls by Aureobasidium pullulans on performance and intestinal morphology in broiler chickens.

    PubMed

    Lai, L P; Lee, M T; Chen, C S; Yu, B; Lee, T T

    2015-12-01

    Soybean hulls are a by-product of soybean processing for oil and meal production; Pleurotus eryngii stalk residues (PESR) are by-products of the edible portion of the fruiting body enriched in bioactive metabolites. This study evaluated the effects of co-fermented PESR and soybean hulls with Aureobasidium pullulans on performance and intestinal morphology in broiler chickens. The in vitro experimental results showed that xylananse and mannanase activity of solid-state fermented soybean hulls (100% SBH) and soybean hulls partially replaced with PESR (75:25, SHP) reached peak at day 12; solid-state fermentation (SSF) enhanced the total phenolic content and trolox equivalency in both products as well. Additionally, FSHP had higher xylotriose and mannobiose levels than fermented FSBH did. A total of 400 broilers (Ross 308) were assigned randomly into four groups receiving the basal diet (control) or the basal diet supplemented with 0.5% fermented SBH (0.5% FSBH), 0.5% fermented SBHP (0.5% FSHP) and 1.0% fermented SBHP (1.0% FSHP) until 35 d of age, respectively. Results demonstrated that 0.5% FSHP addition increased body weight gain as compared with corresponding normal diet fed control in birds during entire experimental period. Compared with the control group, 0.5% FSHP group significantly increased the ratio of lactic acid bacteria to Clostridium perfringens in ceca as well as ileum villus height and jejunum villus height/crypt depth ratio of 35 d old birds. In conclusion, 0.5% FSHP supplementation in the diet could obtain not only improved body weight gain, but optimal intestinal morphology by exerting its bioactive metabolite properties when fed to broilers.

  6. Dendritic cells mediate the induction of polyfunctional human IL17-producing cells (Th17-1 cells) enriched in the bone marrow of patients with myeloma

    PubMed Central

    Barbuto, Scott; Matthews, Phillip; Kukreja, Anjli; Mazumder, Amitabha; Vesole, David; Jagannath, Sundar; Dhodapkar, Madhav V.

    2008-01-01

    IL17-producing (Th17) cells are a distinct lineage of T helper cells that regulate immunity and inflammation. The role of antigen-presenting cells in the induction of Th17 cells in humans remains to be fully defined. Here, we show that human dendritic cells (DCs) are efficient inducers of Th17 cells in culture, including antigen-specific Th17 cells. Although most freshly isolated circulating human Th17 cells secrete IL17 alone or with IL2, those induced by DCs are polyfunctional and coexpress IL17 and IFNγ (Th17-1 cells). The capacity of DCs to expand Th17-1 cells is enhanced upon DC maturation, and mature DCs are superior to monocytes for the expansion of autologous Th17 cells. In myeloma, where tumors are infiltrated by DCs, Th17 cells are enriched in the bone marrow relative to circulation. Bone marrow from patients with myeloma contains a higher proportion of Th17-1 cells compared with the marrow in preneoplastic gammopathy (monoclonal gammopathy of undetermined significance [MGUS]). Uptake of apoptotic but not necrotic myeloma tumor cells by DCs leads to enhanced induction of Th17-1 cells. These data demonstrate the capacity of DCs to induce expansion of polyfunctional IL17-producing T cells in humans, and suggest a role for DCs in the enrichment of Th17-1 cells in the tumor bed. PMID:18669891

  7. Genome sequence of "Candidatus Microthrix parvicella" Bio17-1, a long-chain-fatty-acid-accumulating filamentous actinobacterium from a biological wastewater treatment plant.

    PubMed

    Muller, Emilie E L; Pinel, Nicolás; Gillece, John D; Schupp, James M; Price, Lance B; Engelthaler, David M; Levantesi, Caterina; Tandoi, Valter; Luong, Khai; Baliga, Nitin S; Korlach, Jonas; Keim, Paul S; Wilmes, Paul

    2012-12-01

    "Candidatus Microthrix" bacteria are deeply branching filamentous actinobacteria which occur at the water-air interface of biological wastewater treatment plants, where they are often responsible for foaming and bulking. Here, we report the first draft genome sequence of a strain from this genus: "Candidatus Microthrix parvicella" strain Bio17-1. PMID:23144412

  8. Genome Sequence of “Candidatus Microthrix parvicella” Bio17-1, a Long-Chain-Fatty-Acid-Accumulating Filamentous Actinobacterium from a Biological Wastewater Treatment Plant

    PubMed Central

    Muller, Emilie E. L.; Pinel, Nicolás; Gillece, John D.; Schupp, James M.; Price, Lance B.; Engelthaler, David M.; Levantesi, Caterina; Tandoi, Valter; Luong, Khai; Baliga, Nitin S.; Korlach, Jonas; Keim, Paul S.

    2012-01-01

    “Candidatus Microthrix” bacteria are deeply branching filamentous actinobacteria which occur at the water-air interface of biological wastewater treatment plants, where they are often responsible for foaming and bulking. Here, we report the first draft genome sequence of a strain from this genus: “Candidatus Microthrix parvicella” strain Bio17-1. PMID:23144412

  9. The systematic regulation of oyster CgIL17-1 and CgIL17-5 in response to air exposure.

    PubMed

    Xin, Lusheng; Zhang, Huan; Du, Xinyu; Li, Yiqun; Li, Meijia; Wang, Lingling; Wang, Hao; Qiu, Limei; Song, Linsheng

    2016-10-01

    As a proinflammatory cytokine, vertebrate interleukin 17 (IL17) plays a vital role in the balance of inflammation and homeostasis, and is involved in a systemic regulation of glucose homeostasis. In the present study, a remarkable increase of glucose concentration was observed in oyster serum after 2 d air exposure, which was followed by a rapid up-regulation of CgIL17-1 and CgIL17-5. After oysters was received an injection of extra glucose, the mRNA expressions of CgIL17-1 and CgIL17-5 were also significantly up-regulated. The histopathological changes of hepatopancreas were observed after the oysters were treated by the recombinant proteins of CgIL17-1 and CgIL17-5 in vivo or subjected to air exposure. A significant decrease of GSK3β (Glycogen synthase kinase-3β) protein was also observed after the injection of CgIL17-1 and CgIL17-5 recombinant proteins in vivo. When the oysters with CgIL17-1 and CgIL17-5 genes knocked down were subjected to air exposure, the decline of GSK3β concentration was slowed down and it could still be obviously detected after 7 d compared with that in the control. Meanwhile, the expression of CgDefensin and CgDFFA was inhibited, while CgIAP was up-regulated when CgIL17-1 and CgIL17-5 genes were knocked down, and the oysters exhibited higher mortality (p < 0.05) at 3 d, whereas lower at the late stage of air exposure compared with that in the controls. The results collectively suggested that once oysters were exposed to air, the synthesis of proinflammatory cytokines CgIL17-1 and CgIL17-5 was induced by the up-regulated glucose concentration in oyster serum, which would be not only a negative feedback to the high glucose concentration through mediating the regulation of GSK3β, but also an inducer on tissue damage and immunocompetence as well as the adaptability to stresses. PMID:27268575

  10. A small scale study on the effects of oral administration of the β-glucan produced by Aureobasidium pullulans on milk quality and cytokine expressions of Holstein cows, and on bacterial flora in the intestines of Japanese black calves

    PubMed Central

    2012-01-01

    Background The β–(1 → 3),(1 → 6)-D-glucan extracellularly produced by Aureobasidium pullulans exhibits immunomodulatory activity, and is used for health supplements. To examine the effects of oral administration of the β–(1 → 3),(1 → 6)-D-glucan to domestic animals, a small scale study was conducted using Holstein cows and newborn Japanese Black calves. Findings Holstein cows of which somatic cell count was less than 3 x 105/ml were orally administered with or without the β-(1 → 3),(1 → 6)-D-glucan-enriched A. pullulans cultured fluid (AP-CF) for 3 months, and the properties of milk and serum cytokine expression were monitored. Somatic cell counts were not significantly changed by oral administration of AP-CF, whereas the concentration of solid non fat in the milk tended to increase in the AP-CF administered cows. The results of cytokine expression analysis in the serum using ELISA indicate that the expressions of tumor necrosis factor-α (TNF-α) and interleukin (IL)-6 in all cows which were orally administered with AP-CF became slightly lower than that of control cows after the two-month treatment. On the other hand, IL-8 expression tended to indicate a moderately higher level in all treated cows after the three-month administration of AP-CF in comparison with that of the control cows. Peripartum Japanese Black beef cows and their newborn calves were orally administered with AP-CF, and bacterial flora in the intestines of the calves were analyzed by T-RFLP (terminal restriction fragment length polymorphism). The results suggest that bacterial flora are tendentiously changed by oral administration of AP-CF. Conclusions Our data indicated the possibility that oral administration of the β–(1 → 3),(1 → 6)-D- glucan produced by A. pullulans affects cytokine expressions in the serum of Holstein cows, and influences bacterial flora in the intestines of Japanese Black calves. The findings may be helpful for

  11. Antibody responses against NY-ESO-1 and HER2 antigens in patients vaccinated with combinations of cholesteryl pullulan (CHP)-NY-ESO-1 and CHP-HER2 with OK-432.

    PubMed

    Aoki, Masatoshi; Ueda, Shugo; Nishikawa, Hiroyoshi; Kitano, Shigehisa; Hirayama, Michiko; Ikeda, Hiroaki; Toyoda, Hideki; Tanaka, Kyosuke; Kanai, Michiyuki; Takabayashi, Arimichi; Imai, Hiroshi; Shiraishi, Taizo; Sato, Eiichi; Wada, Hisashi; Nakayama, Eiichi; Takei, Yoshiyuki; Katayama, Naoyuki; Shiku, Hiroshi; Kageyama, Shinichi

    2009-11-16

    Combination vaccines of the NY-ESO-1 protein complexed with cholesteryl pullulan (CHP), CHP-NY-ESO-1, and the truncated 146HER2 protein with CHP, CHP-HER2, were subcutaneously administered with the immuno-adjuvant OK-432 to eight esophageal cancer patients. Vaccination was well-tolerated. NY-ESO-1- and HER2-specific antibody responses were analyzed using the patients' sera and samples from previous single CHP-NY-ESO-1 or CHP-HER2 vaccine trial. The responses to NY-ESO-1 in the combination vaccine study were comparable to the single vaccine. For responses to HER2, there were fewer antibody responses in the combination vaccines. Although there were marked individual variations in the antibody responses to the NY-ESO-1 and HER2 antigens, the reaction patterns to these antigens were comparable within each patient. Antibodies to OK-432 were not augmented. Protein cancer vaccines targeting multiple antigens are feasible.

  12. Micrometastatic cancer cells in bone marrow: in vitro detection with anti-cytokeratin and in vivo labeling with anti-17-1A monoclonal antibodies

    SciTech Connect

    Schlimok, G.; Funke, I.; Holzmann, B.; Goettlinger, G.; Schmidt, G.; Haeser, H.; Swierkot, S.; Warnecke, H.H.; Schneider, B.; Koprowski, H.; Riethmueller, G.

    1987-12-01

    The detection of early micrometastasis or disseminated single tumor cells poses a problem for conventional diagnosis procedures. Using a panel of monoclonal antibodies against cytokeratin and the 17-1A epithelial antigen the authors identified immunocytochemically tumor cells in bone marrow of patients with breast cancer and colorectal cancer at the time of surgery of the primary tumor. Monoclonal antibody CK2, recognizing the human cytokeratin component 18 in simple epithelia, appeared to be the most suitable reagent because of its negative reaction with bone marrow samples of the noncarcinoma patients. Its specificity was further demonstrated in a double-marker staining procedure using an anti-leukocyte common antigen monoclonal antibody (T200) as counterstain. A comparative analysis showed that immunocytology was clearly superior to conventional cytology and histology. In 9.5-20.5% of patients without distant metastasis, tumor cells could be detected in bone marrow. They found a significant correlation between tumor cells in bone marrow and conventional risk factors, such as distant metastasis or lymph node involvement. In a first approach toward immunotherapy they demonstrated in 3 patients that infused monoclonal antibody 17-1A can label single tumor cells in bone marrow in vivo. They then used this single approach to follow up on 7 patients undergoing 17-1A therapy in an adjuvant clinical trial.

  13. Correlation of maple sap composition with bacterial and fungal communities determined by multiplex automated ribosomal intergenic spacer analysis (MARISA).

    PubMed

    Filteau, Marie; Lagacé, Luc; LaPointe, Gisèle; Roy, Denis

    2011-08-01

    During collection, maple sap is contaminated by bacteria and fungi that subsequently colonize the tubing system. The bacterial microbiota has been more characterized than the fungal microbiota, but the impact of both components on maple sap quality remains unclear. This study focused on identifying bacterial and fungal members of maple sap and correlating microbiota composition with maple sap properties. A multiplex automated ribosomal intergenic spacer analysis (MARISA) method was developed to presumptively identify bacterial and fungal members of maple sap samples collected from 19 production sites during the tapping period. Results indicate that the fungal community of maple sap is mainly composed of yeast related to Mrakia sp., Mrakiella sp., Guehomyces pullulans, Cryptococcus victoriae and Williopsis saturnus. Mrakia, Mrakiella and Guehomyces peaks were identified in samples of all production sites and can be considered dominant and stable members of the fungal microbiota of maple sap. A multivariate analysis based on MARISA profiles and maple sap chemical composition data showed correlations between Candida sake, Janthinobacterium lividum, Williopsis sp., Leuconostoc mesenteroides, Mrakia sp., Rhodococcus sp., Pseudomonas tolaasii, G. pullulans and maple sap composition at different flow periods. This study provides new insights on the relationship between microbial community and maple sap quality.

  14. Growth suppression of colorectal cancer by plant-derived multiple mAb CO17-1A × BR55 via inhibition of ERK1/2 phosphorylation.

    PubMed

    Kwak, Dong Hoon; Moussavou, Ghislain; Lee, Ju Hyoung; Heo, Sung Youn; Ko, Kisung; Hwang, Kyung-A; Jekal, Seung-Joo; Choo, Young-Kug

    2014-01-01

    We have generated the transgenic Tabaco plants expressing multiple monoclonal antibody (mAb) CO7-1A × BR55 by cross-pollinating with mAb CO17-1A and mAb BR55. We have demonstrated the anti-cancer effect of plant-derived multiple mAb CO17-1A × BR55. We find that co-treatment of colorectal mAbs (anti-epithelial cellular adhesion molecule (EpCAM), plant-derived monoclonal antibody (mAb(P)) CO17-1A and mAb(P) CO17-1A × BR55) with RAW264.7 cells significantly inhibited the cell growth in SW620 cancer cells. In particular, multi mAb(P) CO17-1A × BR55 significantly and efficiently suppressed the growth of SW620 cancer cells compared to another mAbs. Apoptotic death-positive cells were significantly increased in the mAb(P) CO17-1A × BR55-treated. The mAb(P) CO17-1A × BR55 treatment significantly decreased the expression of B-Cell lymphoma-2 (BCl-2), but the expression of Bcl-2-associated X protein (Bax), and cleaved caspase-3 were markedly increased. In vivo, the mAb(P) CO17-1A × BR55 significantly and efficiently inhibited the growth of colon tumors compared to another mAbs. The apoptotic cell death and inhibition of pro-apoptotic proteins expression were highest by treatment with mAb(P) CO17-1A × BR55. In addition, the mAb(P) CO17-1A × BR55 significantly inhibited the extracellular signal-regulated kinase 1 and 2 (ERK1/2) phosphorylation in cancer cells and tumors. Therefore, this study results suggest that multiple mAb(P) CO17-1A × BR55 has a significant effect on apoptosis-mediated anticancer by suppression of ERK1/2 phosphorylation in colon cancer compared to another mAbs. In light of these results, further clinical investigation should be conducted on mAb(P) CO17-1A × BR55 to determine its possible chemopreventive and/or therapeutic efficacy against human colon cancer. PMID:25405740

  15. Growth suppression of colorectal cancer by plant-derived multiple mAb CO17-1A × BR55 via inhibition of ERK1/2 phosphorylation.

    PubMed

    Kwak, Dong Hoon; Moussavou, Ghislain; Lee, Ju Hyoung; Heo, Sung Youn; Ko, Kisung; Hwang, Kyung-A; Jekal, Seung-Joo; Choo, Young-Kug

    2014-11-14

    We have generated the transgenic Tabaco plants expressing multiple monoclonal antibody (mAb) CO7-1A × BR55 by cross-pollinating with mAb CO17-1A and mAb BR55. We have demonstrated the anti-cancer effect of plant-derived multiple mAb CO17-1A × BR55. We find that co-treatment of colorectal mAbs (anti-epithelial cellular adhesion molecule (EpCAM), plant-derived monoclonal antibody (mAb(P)) CO17-1A and mAb(P) CO17-1A × BR55) with RAW264.7 cells significantly inhibited the cell growth in SW620 cancer cells. In particular, multi mAb(P) CO17-1A × BR55 significantly and efficiently suppressed the growth of SW620 cancer cells compared to another mAbs. Apoptotic death-positive cells were significantly increased in the mAb(P) CO17-1A × BR55-treated. The mAb(P) CO17-1A × BR55 treatment significantly decreased the expression of B-Cell lymphoma-2 (BCl-2), but the expression of Bcl-2-associated X protein (Bax), and cleaved caspase-3 were markedly increased. In vivo, the mAb(P) CO17-1A × BR55 significantly and efficiently inhibited the growth of colon tumors compared to another mAbs. The apoptotic cell death and inhibition of pro-apoptotic proteins expression were highest by treatment with mAb(P) CO17-1A × BR55. In addition, the mAb(P) CO17-1A × BR55 significantly inhibited the extracellular signal-regulated kinase 1 and 2 (ERK1/2) phosphorylation in cancer cells and tumors. Therefore, this study results suggest that multiple mAb(P) CO17-1A × BR55 has a significant effect on apoptosis-mediated anticancer by suppression of ERK1/2 phosphorylation in colon cancer compared to another mAbs. In light of these results, further clinical investigation should be conducted on mAb(P) CO17-1A × BR55 to determine its possible chemopreventive and/or therapeutic efficacy against human colon cancer.

  16. Growth Suppression of Colorectal Cancer by Plant-Derived Multiple mAb CO17-1A × BR55 via Inhibition of ERK1/2 Phosphorylation

    PubMed Central

    Kwak, Dong Hoon; Moussavou, Ghislain; Lee, Ju Hyoung; Heo, Sung Youn; Ko, Kisung; Hwang, Kyung-A; Jekal, Seung-Joo; Choo, Young-Kug

    2014-01-01

    We have generated the transgenic Tabaco plants expressing multiple monoclonal antibody (mAb) CO7-1A × BR55 by cross-pollinating with mAb CO17-1A and mAb BR55. We have demonstrated the anti-cancer effect of plant-derived multiple mAb CO17-1A × BR55. We find that co-treatment of colorectal mAbs (anti-epithelial cellular adhesion molecule (EpCAM), plant-derived monoclonal antibody (mAbP) CO17-1A and mAbP CO17-1A × BR55) with RAW264.7 cells significantly inhibited the cell growth in SW620 cancer cells. In particular, multi mAbP CO17-1A × BR55 significantly and efficiently suppressed the growth of SW620 cancer cells compared to another mAbs. Apoptotic death-positive cells were significantly increased in the mAbP CO17-1A × BR55-treated. The mAbP CO17-1A × BR55 treatment significantly decreased the expression of B-Cell lymphoma-2 (BCl-2), but the expression of Bcl-2-associated X protein (Bax), and cleaved caspase-3 were markedly increased. In vivo, the mAbP CO17-1A × BR55 significantly and efficiently inhibited the growth of colon tumors compared to another mAbs. The apoptotic cell death and inhibition of pro-apoptotic proteins expression were highest by treatment with mAbP CO17-1A × BR55. In addition, the mAbP CO17-1A × BR55 significantly inhibited the extracellular signal-regulated kinase 1 and 2 (ERK1/2) phosphorylation in cancer cells and tumors. Therefore, this study results suggest that multiple mAbP CO17-1A × BR55 has a significant effect on apoptosis-mediated anticancer by suppression of ERK1/2 phosphorylation in colon cancer compared to another mAbs. In light of these results, further clinical investigation should be conducted on mAbP CO17-1A × BR55 to determine its possible chemopreventive and/or therapeutic efficacy against human colon cancer. PMID:25405740

  17. Yeasts from sub-Antarctic region: biodiversity, enzymatic activities and their potential as oleaginous microorganisms.

    PubMed

    Martinez, A; Cavello, I; Garmendia, G; Rufo, C; Cavalitto, S; Vero, S

    2016-09-01

    Various microbial groups are well known to produce a range of extracellular enzymes and other secondary metabolites. However, the occurrence and importance of investment in such activities have received relatively limited attention in studies of Antarctic soil microbiota. Sixty-one yeasts strains were isolated from King George Island, Antarctica which were characterized physiologically and identified at the molecular level using the D1/D2 region of rDNA. Fifty-eight yeasts (belonging to the genera Cryptococcus, Leucosporidiella, Rhodotorula, Guehomyces, Candida, Metschnikowia and Debaryomyces) were screened for extracellular amylolytic, proteolytic, esterasic, pectinolytic, inulolytic xylanolytic and cellulolytic activities at low and moderate temperatures. Esterase activity was the most common enzymatic activity expressed by the yeast isolates regardless the assay temperature and inulinase was the second most common enzymatic activity. No cellulolytic activity was detected. One yeast identified as Guehomyces pullulans (8E) showed significant activity across six of seven enzymes types tested. Twenty-eight yeast isolates were classified as oleaginous, being the isolate 8E the strain that accumulated the highest levels of saponifiable lipids (42 %). PMID:27469174

  18. The influence of Aster x salignus Willd. Invasion on the diversity of soil yeast communities

    NASA Astrophysics Data System (ADS)

    Glushakova, A. M.; Kachalkin, A. V.; Chernov, I. Yu.

    2016-07-01

    The annual dynamics of yeast communities were studied in the soddy-podzolic soil under the thickets of Aster x salignus Willd., one of the widespread invasive plant species in central Russia. Yeast groups in the soils under continuous aster thickets were found to differ greatly from the yeast communities in the soils under the adjacent indigenous meadow vegetation. In both biotopes the same species ( Candida vartiovaarae, Candida sake, and Cryptococcus terreus) are dominants. However, in the soils under indigenous grasses, eurybiontic yeasts Rhodotorula mucilaginosa, which almost never occur in the soil under aster, are widespread. In the soil under aster, the shares of other typical epiphytic and pedobiontic yeast fungi (ascomycetic species Wickerhamomyces aniomalus, Barnettozyma californica and basidiomycetic species Cystofilobasidium macerans, Guehomyces pullulans) significantly increase. Thus, the invasion of Aster x salignus has a clear effect on soil yeast complexes reducing their taxonomic and ecological diversity.

  19. Histological improvement in salivary gland along with effector memory Th17-1 cell reduction in a primary Sjogren's syndrome patient with dermatomyositis and diffuse large B-cell lymphoma by R-CHOP therapy.

    PubMed

    Koga, Tomohiro; Mizokami, Akinari; Nakashima, Masahiro; Shimizu, Toshimasa; Nakashima, Yoshikazu; Nakamura, Hideki; Chiwata, Masahiko; Daisuke, Niino; Kawakami, Atsushi

    2016-04-01

    We treated a 45-year-old Japanese woman with primary Sjogren's syndrome (SS) complicated with dermatomyositis (DM) followed by diffuse large B-cell lymphoma. She was admitted to our hospital for further evaluation of fever, weight loss and peritoneal lymphadenopathy. The histological examination of her lymph node revealed diffuse large B-cell lymphoma. The patient was then treated with 8 cycles of R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone) therapy, which resulted in histological and functional improvement of her salivary glands. Of note, the percentage of effector memory Th17-1 (CD3+CD4+CD45RA-CCR7-CXCR3+CCR6+) cells in the peripheral blood was decreased after the R-CHOP treatment. This case suggests that an altered Th17-1 cell subset by B-cell depletion therapy is critical for the improvement of tissue damage in patients with SS, and the case suggests that clinicians should consider measuring the effector memory Th-subsets to predict the disease activity in SS patients. PMID:26960952

  20. 21 CFR 17.1 - Scope.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL CIVIL MONEY PENALTIES HEARINGS... violations of the act relating to postmarket studies, clinical trial requirements, and risk evaluation and...) Section 351(d)(2) of the Public Health Service Act (the PHS Act) authorizing civil money penalties...

  1. 21 CFR 17.1 - Scope.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL CIVIL MONEY PENALTIES HEARINGS... violations of the act relating to postmarket studies, clinical trial requirements, and risk evaluation and...) Section 351(d)(2) of the Public Health Service Act (the PHS Act) authorizing civil money penalties...

  2. 21 CFR 17.1 - Scope.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL CIVIL MONEY PENALTIES HEARINGS... violations of the act relating to postmarket studies, clinical trial requirements, and risk evaluation and...) Section 351(d)(2) of the Public Health Service Act (the PHS Act) authorizing civil money penalties...

  3. 21 CFR 17.1 - Scope.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL CIVIL MONEY PENALTIES HEARINGS... violations of the act relating to postmarket studies, clinical trial requirements, and risk evaluation and...) Section 351(d)(2) of the Public Health Service Act (the PHS Act) authorizing civil money penalties...

  4. 21 CFR 17.1 - Scope.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... violations relating to the submission of certifications and/or clinical trial information to the clinical trial data bank and section 303(f)(4) of the act authorizing civil money penalties for certain violations of the act relating to postmarket studies, clinical trial requirements, and risk evaluation...

  5. 7 CFR 17.1 - General.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... the United States and have a person, principal, or agent on whom service of judicial process may be... assurances of repayment to CCC for the financing extended by CCC. (c) Purchase authorizations. This part covers, among other things, the issuance by the General Sales Manager of purchase authorizations...

  6. 15 CFR 17.1 - Licensing rules.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... patents and patent applications vested in the United States of America, found at 41 CFR 101-4.1, are... clarifications: (1) The term “Government agency” as defined at 41 CFR 101-4.102(c) means the United States... Government agency”, as defined at 41 CFR 101-4.102(d), means the Secretary of Commerce or a designee. (b)...

  7. 15 CFR 17.1 - Licensing rules.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... patents and patent applications vested in the United States of America, found at 41 CFR 101-4.1, are... clarifications: (1) The term “Government agency” as defined at 41 CFR 101-4.102(c) means the United States... Government agency”, as defined at 41 CFR 101-4.102(d), means the Secretary of Commerce or a designee. (b)...

  8. 15 CFR 17.1 - Licensing rules.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... patents and patent applications vested in the United States of America, found at 41 CFR 101-4.1, are... clarifications: (1) The term “Government agency” as defined at 41 CFR 101-4.102(c) means the United States... Government agency”, as defined at 41 CFR 101-4.102(d), means the Secretary of Commerce or a designee. (b)...

  9. 15 CFR 17.1 - Licensing rules.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... patents and patent applications vested in the United States of America, found at 41 CFR 101-4.1, are... clarifications: (1) The term “Government agency” as defined at 41 CFR 101-4.102(c) means the United States... Government agency”, as defined at 41 CFR 101-4.102(d), means the Secretary of Commerce or a designee. (b)...

  10. 45 CFR 17.1 - Definition.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL ADMINISTRATION RELEASE OF ADVERSE INFORMATION TO NEWS MEDIA... Department or any principal operating component made to the news media inviting public attention to an action... affect persons or organizations identified therein. This part does not apply to nor is it affected by...

  11. 32 CFR 17.1 - Purpose.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... conduct of trials by a military commission appointed pursuant to 32 CFR part 9 and Military Order of November 13, 2001, “Detention, Treatment, and Trial of Certain Non-Citizens in the War Against Terrorism,” (3 CFR 2001 Comp., p. 918, 66 FR 57833)....

  12. 45 CFR 17.1 - Definition.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... Regulation (45 CFR part 5). ... DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL ADMINISTRATION RELEASE OF ADVERSE INFORMATION TO NEWS MEDIA... Department or any principal operating component made to the news media inviting public attention to an...

  13. Spectroscopic characterization and molecular structure of 3,14-dimethyl-2,6,13,17-tetraazapentacyclo[16.4.0.1(2,17).1(6,13).0(7,12)]tetracosane.

    PubMed

    Moon, Dohyun; Hong, Yong Pyo; Choi, Jong Ha

    2016-09-01

    Constrained cyclam derivatives have been found to exhibit anti-HIV effects. The strength of binding to the CXCR4 receptor correlates with anti-HIV activity. The conformation of the macrocyclic compound is very important for co-receptor recognition. Therefore, knowledge of the conformation and crystal packing of macrocycles has become important in developing new highly effective anti-HIV drugs. Structural modifications of N-functionalized polyaza macrocyclic compounds have been achieved using various methods. A new synthesis affording single crystals of the title tetraazapentacyclo[16.4.0.1(2,17).1(6,13).0(7,12)]tetracosane macrocycle, C22H40N4, is reported. Formaldehyde reacts readily at room temperature with the tetraazatricyclo[16.4.0.0(2,17)]docosane precursor to yield a macropolycycle containing two five-membered rings. Characterization by elemental, spectroscopic and single-crystal X-ray diffraction analyses shows that the asymmetric unit contains half of a centrosymmetric molecule. The molecular structure shows a trans conformation for the two methylene bridges owing to molecular symmetry. The crystal structure is stabilized by intramolecular C-H...N hydrogen bonds. NMR and IR spectroscopic properties support the methylene-bridged macrocyclic structure. PMID:27585935

  14. Maple sap predominant microbial contaminants are correlated with the physicochemical and sensorial properties of maple syrup.

    PubMed

    Filteau, Marie; Lagacé, Luc; Lapointe, Gisèle; Roy, Denis

    2012-03-01

    Maple sap processing and microbial contamination are significant aspects that affect maple syrup quality. In this study, two sample sets from 2005 and 2008 were used to assess the maple syrup quality variation and its relationship to microbial populations, with respect to processing, production site and harvesting period. The abundance of maple sap predominant bacteria (Pseudomonas fluorescens group and two subgroups, Rahnella spp., Janthinobacterium spp., Leuconostoc mesenteroides) and yeast (Mrakia spp., Mrakiella spp.,Guehomyces pullulans) was assessed by quantitative PCR. Maple syrup properties were analyzed by physicochemical and sensorial methods. Results indicate that P. fluorescens, Mrakia spp., Mrakiella spp. G. pullulans and Rahnella spp. are stable contaminants of maple sap, as they were found for every production site throughout the flow period. Multiple factor analysis reports a link between the relative abundance of P. fluorescens group and Mrakia spp. in maple sap with maple and vanilla odor as well as flavor of maple syrup. This evidence supports the contribution of these microorganisms or a consortium of predominant microbial contaminants to the characteristic properties of maple syrup.

  15. Cyclostratigraphy and eccentricity tuning of the early Oligocene through early Miocene (30.1-17.1 Ma): Cibicides mundulus stable oxygen and carbon isotope records from Walvis Ridge Site 1264

    NASA Astrophysics Data System (ADS)

    Liebrand, Diederik; Beddow, Helen M.; Lourens, Lucas J.; Pälike, Heiko; Raffi, Isabella; Bohaty, Steven M.; Hilgen, Frederik J.; Saes, Mischa J. M.; Wilson, Paul A.; van Dijk, Arnold E.; Hodell, David A.; Kroon, Dick; Huck, Claire E.; Batenburg, Sietske J.

    2016-09-01

    Few astronomically calibrated high-resolution (≤5 kyr) climate records exist that span the Oligocene-Miocene time interval. Notably, available proxy records show responses varying in amplitude at frequencies related to astronomical forcing, and the main pacemakers of global change on astronomical time-scales remain debated. Here we present newly generated X-ray fluorescence core scanning and benthic foraminiferal stable oxygen and carbon isotope records from Ocean Drilling Program Site 1264 (Walvis Ridge, southeastern Atlantic Ocean). Complemented by data from nearby Site 1265, the Site 1264 benthic stable isotope records span a continuous ∼13-Myr interval of the Oligo-Miocene (30.1-17.1 Ma) at high resolution (∼3.0 kyr). Spectral analyses in the stratigraphic depth domain indicate that the largest amplitude variability of all proxy records is associated with periods of ∼ 3.4 m and ∼ 0.9 m, which correspond to 405- and ∼110-kyr eccentricity, using a magnetobiostratigraphic age model. Maxima in CaCO3 content, δ18O and δ13C are interpreted to coincide with ∼110 kyr eccentricity minima. The strong expression of these cycles in combination with the weakness of the precession- and obliquity-related signals allow construction of an astronomical age model that is solely based on tuning the CaCO3 content to the nominal (La2011_ecc3L) eccentricity solution. Very long-period eccentricity maxima (∼2.4-Myr) are marked by recurrent episodes of high-amplitude ∼110-kyr δ18O cycles at Walvis Ridge, indicating greater sensitivity of the climate/cryosphere system to short eccentricity modulation of climatic precession. In contrast, the responses of the global (high-latitude) climate system, cryosphere, and carbon cycle to the 405-kyr cycle, as expressed in benthic δ18O and especially δ13C signals, are more pronounced during ∼2.4-Myr minima. The relationship between the recurrent episodes of high-amplitude ∼110-kyr δ18O cycles and the ∼1.2-Myr amplitude

  16. 38 CFR 17.1 - Incorporation by reference.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 1 CFR part 51. To enforce an edition of a publication other than that specified in this section, VA will provide notice of the change in a notice of proposed rulemaking in the Federal Register and the... Monoxide (CO) Detection and Warning Equipment (2009 edition), IBR approved for § 17.74. (Authority: 5...

  17. 38 CFR 17.1 - Incorporation by reference.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 1 CFR part 51. To enforce an edition of a publication other than that specified in this section, VA will provide notice of the change in a notice of proposed rulemaking in the Federal Register and the... Monoxide (CO) Detection and Warning Equipment (2009 edition), IBR approved for § 17.74. (Authority: 5...

  18. 38 CFR 17.1 - Incorporation by reference.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 1 CFR part 51. To enforce an edition of a publication other than that specified in this section, VA... information on the availability of approved materials at NARA, call (202) 741-6030, or go to:...

  19. 47 CFR 17.1 - Basis and purpose.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... Telecommunication FEDERAL COMMUNICATIONS COMMISSION GENERAL CONSTRUCTION, MARKING, AND LIGHTING OF ANTENNA..., and/or illumination of antenna structures if and when in its judgment such structures constitute, or... this part is to prescribe certain procedures for antenna structure registration and standards...

  20. 47 CFR 17.1 - Basis and purpose.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... Telecommunication FEDERAL COMMUNICATIONS COMMISSION GENERAL CONSTRUCTION, MARKING, AND LIGHTING OF ANTENNA..., and/or illumination of antenna structures if and when in its judgment such structures constitute, or... this part is to prescribe certain procedures for antenna structure registration and standards...

  1. 38 CFR 17.1 - Incorporation by reference.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 1 CFR part 51. To enforce an edition of a publication other than that specified in this section, VA... this part. (1) NFPA 10, Standard for Portable Fire Extinguishers (2010 edition), Incorporation by Reference (IBR) approved for §§ 17.63, 17.74, and 17.81. (2) NFPA 101, Life Safety Code (2009 edition),...

  2. 50 CFR 17.1 - Purpose of regulations.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... Wildlife and Fisheries UNITED STATES FISH AND WILDLIFE SERVICE, DEPARTMENT OF THE INTERIOR (CONTINUED... PLANTS (CONTINUED) ENDANGERED AND THREATENED WILDLIFE AND PLANTS Introduction and General Provisions § 17... provided in part 23 of this subchapter. (b) The regulations identify those species of wildlife and...

  3. 50 CFR 17.1 - Purpose of regulations.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... Wildlife and Fisheries UNITED STATES FISH AND WILDLIFE SERVICE, DEPARTMENT OF THE INTERIOR (CONTINUED... PLANTS (CONTINUED) ENDANGERED AND THREATENED WILDLIFE AND PLANTS Introduction and General Provisions § 17... provided in part 23 of this subchapter. (b) The regulations identify those species of wildlife and...

  4. 50 CFR 17.1 - Purpose of regulations.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... Wildlife and Fisheries UNITED STATES FISH AND WILDLIFE SERVICE, DEPARTMENT OF THE INTERIOR (CONTINUED... PLANTS (CONTINUED) ENDANGERED AND THREATENED WILDLIFE AND PLANTS Introduction and General Provisions § 17... provided in part 23 of this subchapter. (b) The regulations identify those species of wildlife and...

  5. 50 CFR 17.1 - Purpose of regulations.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... Wildlife and Fisheries UNITED STATES FISH AND WILDLIFE SERVICE, DEPARTMENT OF THE INTERIOR (CONTINUED... PLANTS (CONTINUED) ENDANGERED AND THREATENED WILDLIFE AND PLANTS Introduction and General Provisions § 17... provided in part 23 of this subchapter. (b) The regulations identify those species of wildlife and...

  6. 50 CFR 17.1 - Purpose of regulations.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... Wildlife and Fisheries UNITED STATES FISH AND WILDLIFE SERVICE, DEPARTMENT OF THE INTERIOR (CONTINUED... PLANTS (CONTINUED) ENDANGERED AND THREATENED WILDLIFE AND PLANTS Introduction and General Provisions § 17... provided in part 23 of this subchapter. (b) The regulations identify those species of wildlife and...

  7. 27 CFR 17.1 - Scope of regulations.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... manufacture of medicines, medicinal preparations, food products, flavors, flavoring extracts, and perfume that... nonbeverage products; and bonds, claims, formulas and samples, losses, and records to be kept pertaining...

  8. Yeast and yeast-like diversity in the southernmost glacier of Europe (Calderone Glacier, Apennines, Italy).

    PubMed

    Branda, Eva; Turchetti, Benedetta; Diolaiuti, Guglielmina; Pecci, Massimo; Smiraglia, Claudio; Buzzini, Pietro

    2010-06-01

    The present study reports the characterization of psychrophilic yeast and yeast-like diversity in cold habitats (superficial and deep sediments, ice cores and meltwaters) of the Calderone Glacier (Italy), which is the southernmost glacier in Europe. After incubation at 4 and 20 degrees C, sediments contained about 10(2)-10(3) CFU of yeasts g(-1). The number of viable yeast cells in ice and meltwaters was several orders of magnitude lower. The concomitant presence of viable bacteria and filamentous fungi has also been observed. In all, 257 yeast strains were isolated and identified by 26S rRNA gene D1/D2 and internal transcribed spacers (1 and 2) sequencing as belonging to 28 ascomycetous and basidiomycetous species of 11 genera (Candida, Cystofilobasidium, Cryptococcus, Dioszegia, Erythrobasidium, Guehomyces, Mastigobasidium, Mrakia, Mrakiella, Rhodotorula and Sporobolomyces). Among them, the species Cryptococcus gastricus accounted for almost 40% of the total isolates. In addition, 12 strains were identified as belonging to the yeast-like species Aureobasidium pullulans and Exophiala dermatitidis, whereas 15 strains, presumably belonging to new species, yet to be described, were also isolated. Results herein reported indicate that the Calderone Glacier, although currently considered a vanishing ice body due to the ongoing global-warming phenomenon, still harbors viable psychrophilic yeast populations. Differences of yeast and yeast-like diversity between the glacier under study and other worldwide cold habitats are also discussed.

  9. 26 CFR 1.401(a)(17)-1 - Limitation on annual compensation.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... effective date, annual compensation limit means $200,000, adjusted as provided by the Commissioner. The... first plan year beginning on or after the statutory effective date (generally $200,000) must be applied... for a 12-consecutive-month period, or periods, ending no later than the last day of the plan...

  10. 26 CFR 1.401(a)(17)-1 - Limitation on annual compensation.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... effective date, annual compensation limit means $200,000, adjusted as provided by the Commissioner. The... first plan year beginning on or after the statutory effective date (generally $200,000) must be applied... for a 12-consecutive-month period, or periods, ending no later than the last day of the plan...

  11. 26 CFR 1.401(a)(17)-1 - Limitation on annual compensation.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... limit means $200,000, adjusted as provided by the Commissioner. The amount of the annual compensation... statutory effective date (generally $200,000) must be applied to compensation for that prior plan year. (3... periods, ending no later than the last day of the plan year, then the annual compensation limit applies...

  12. 26 CFR 31.3306(c)(17)-1 - Fishing services.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... individual in the catching, taking, harvesting, cultivating, or farming of any kind of fish, shell-fish (for... cleaning, icing, and packing of fish as are necessary for the immediate preservation of the catch....

  13. Ecology and Oil Spills, 17-1. Military Curriculum Materials for Vocational and Technical Education.

    ERIC Educational Resources Information Center

    Army Quartermaster School, Ft. Lee, VA.

    This single-lesson course in ecology and oil spills for the secondary/postsecondary level comprises one of a number of military-developed curriculum packages selected for adaptation to vocational instruction and curriculum development in a civilian setting. The purpose stated for the 2-hour course is to provide students with an understanding of…

  14. Millwright Apprenticeship. Related Training Modules. 17.1-17.13 Hydraulics.

    ERIC Educational Resources Information Center

    Lane Community Coll., Eugene, OR.

    This packet of 13 learning modules on hydraulics is 1 of 6 such packets developed for apprenticeship training for millwrights. Introductory materials are a complete listing of all available modules and a supplementary reference list. Each module contains some or all of these components: goal, performance indicators, study guide (a check list of…

  15. 26 CFR 1.381(c)(17)-1 - Deficiency dividend of personal holding company.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 4 2010-04-01 2010-04-01 false Deficiency dividend of personal holding company... Deficiency dividend of personal holding company. (a) Carryover requirement. If a determination (as defined in... 381(a) applies is liable for personal holding company tax imposed by section 541 (or by...

  16. 26 CFR 1.401(a)(17)-1 - Limitation on annual compensation.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... beginning on or after January 1, 1996, of the governing body with authority to amend the plan, if that body does not meet continuously. For purposes of this paragraph (d)(4), the term governing body with authority to amend the plan means the legislature, board, commission, council, or other governing body...

  17. METRO-APEX Volume 17.1: Industrialist's Manual No. 7, Shick Cannery. Revised.

    ERIC Educational Resources Information Center

    University of Southern California, Los Angeles. COMEX Research Project.

    The Industrialist's Manual No. 7 (Shick Cannery) is one of a set of twenty-one manuals used in METRO-APEX 1974, a computerized college and professional level, computer-supported, role-play, simulation exercise of a community with "normal" problems. Stress is placed on environmental quality considerations. APEX 1974 is an expansion of APEX--Air…

  18. 26 CFR 1.501(c)(17)-1 - Supplemental unemployment benefit trusts.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 Internal Revenue 7 2013-04-01 2013-04-01 false Supplemental unemployment benefit trusts. 1.501... Supplemental unemployment benefit trusts. (a) Requirements for qualification. (1) A supplemental unemployment... the purpose of providing supplemental unemployment compensation benefits (as defined in section...

  19. 26 CFR 1.501(c)(17)-1 - Supplemental unemployment benefit trusts.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 7 2010-04-01 2010-04-01 true Supplemental unemployment benefit trusts. 1.501(c... Supplemental unemployment benefit trusts. (a) Requirements for qualification. (1) A supplemental unemployment... the purpose of providing supplemental unemployment compensation benefits (as defined in section...

  20. 26 CFR 1.501(c)(17)-1 - Supplemental unemployment benefit trusts.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 Internal Revenue 7 2012-04-01 2012-04-01 false Supplemental unemployment benefit trusts. 1.501... Supplemental unemployment benefit trusts. (a) Requirements for qualification. (1) A supplemental unemployment... the purpose of providing supplemental unemployment compensation benefits (as defined in section...

  1. 26 CFR 1.501(c)(17)-1 - Supplemental unemployment benefit trusts.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 7 2011-04-01 2009-04-01 true Supplemental unemployment benefit trusts. 1.501(c... Supplemental unemployment benefit trusts. (a) Requirements for qualification. (1) A supplemental unemployment... the purpose of providing supplemental unemployment compensation benefits (as defined in section...

  2. 26 CFR 1.501(c)(17)-1 - Supplemental unemployment benefit trusts.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 7 2014-04-01 2013-04-01 true Supplemental unemployment benefit trusts. 1.501(c... Supplemental unemployment benefit trusts. (a) Requirements for qualification. (1) A supplemental unemployment... the purpose of providing supplemental unemployment compensation benefits (as defined in section...

  3. 26 CFR 31.3306(c)(17)-1 - Fishing services.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...) Salmon and halibut fishing. Services performed in connection with the catching or taking of salmon or... taking of salmon or halibut, for commercial purposes, nor the services of any other individual...

  4. 26 CFR 31.3306(c)(17)-1 - Fishing services.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...) Salmon and halibut fishing. Services performed in connection with the catching or taking of salmon or... taking of salmon or halibut, for commercial purposes, nor the services of any other individual...

  5. 26 CFR 31.3306(c)(17)-1 - Fishing services.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...) Salmon and halibut fishing. Services performed in connection with the catching or taking of salmon or... taking of salmon or halibut, for commercial purposes, nor the services of any other individual...

  6. 26 CFR 31.3306(c)(17)-1 - Fishing services.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...) Salmon and halibut fishing. Services performed in connection with the catching or taking of salmon or... taking of salmon or halibut, for commercial purposes, nor the services of any other individual...

  7. 26 CFR 1.401(a)(17)-1 - Limitation on annual compensation.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    .... Paragraph (d) of this section provides the effective dates of section 401(a)(17), the amendments made by...) of this section provides rules for determining post-effective-date accrued benefits under the fresh... of this section, for plan years beginning prior to the OBRA '93 effective date, annual...

  8. EXPRESSION OF GREEN FLUORESCENT PROTEIN IN AUREOBASIDIUM PULLULANS AND QUANTIFICATION OF THE FUNGUS ON LEAF SURFACES. (R823845)

    EPA Science Inventory

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  9. High molecular weight β-poly(L-malic acid) produced by A. pullulans with Ca²⁺ added repeated batch culture.

    PubMed

    Cao, Weifeng; Chen, Xiangrong; Luo, Jianquan; Yin, Junxiang; Qiao, Changsheng; Wan, Yinhua

    2016-04-01

    β-Poly(malic acid) (PMLA) has attracted increasing attentions because of its potential application in medicine and other industries. In this study, the variation of PMLA molecular weight (Mw) in the batch culture and the strategies to enhance PMLA Mw were studied. Adding exogenous Ca(2+) (0.1g/L CaCl2) to the medium caused a significant increase in both PMLA concentration and Mw (11.38% and 26.3%, respectively) when Na2CO3 was used as the neutralizer. The Mw of PMLA during the process of batch culture, which associated with the specific PMLA production per unit cell mass (Yp/x) before glucose was depleted, increased from 12.522 KDa to its maximum 18.693 KDa and then kept decreasing until the end of the culture. Compared with the results in batch culture, Mw increased by 84.4% (up to 19.51 kDa) with a productivity of 1.1 gh(-1)L(-1) when the cells were maintained in exponential growth phase during Ca(2+) added repeated batch culture. The present work provides an efficient approach to obtain superior quality PMLA product with high Mw.

  10. Covalent Immobilization and Characterization of a Novel Pullulanase from Fontibacillus sp. Strain DSHK 107 onto Florisil® and Nano-silica for Pullulan Hydrolysis.

    PubMed

    Alagöz, Dilek; Yildirim, Deniz; Güvenmez, Hatice Korkmaz; Sihay, Damla; Tükel, S Seyhan

    2016-08-01

    A novel pullulanase partially purified from Fontibacillus sp. was covalently immobilized on Florisil® and nano-silica through both glutaraldehyde and (3-glycidyloxypropyl)trimethoxysilane spacer arms. The pullulanase immobilized on Florisil® and nano-silica through glutaraldehyde spacer arm showed 85 and 190 % activity of its free form, respectively, whereas no activity was observed when it was immobilized on the same supports through (3-glycidyloxypropyl)trimethoxysilane spacer arm. The maximum working pHs of both the immobilized pullulanases on Florisil® and nano-silica through glutaraldehyde spacer arm were determined as 5.0; however, the maximum working pH of the free pullulanase was pH 6.0. The maximum temperatures of all the pullulanase preparations were determined as 35 °C. The apparent K m values were 1.49, 1.54, and 0.59 mg/mL pullunan, respectively, for the free and immobilized pullulanases on Florisil® and nano-silica. The corresponding apparent V max values were 0.59, 1.53, and 1.57 U mg prot.(-1) min.(-1). Thermal stability of pullulanases immobilized on Florisil® and nano-silica was enhanced 6.5- and 15.6-folds, respectively at 35 °C and 6.6- and 16.0-folds, respectively, at 50 °C. The pullulanases immobilized on Florisil® and nano-silica protected 71 and 90 % of their initial activities after 10 reuses.

  11. Development of a stable cobalt-ruthenium Fischer-Tropsch catalyst. Technical progress report No. 17, 1 November 1993--31 December 1993

    SciTech Connect

    Frame, R.R.; Gala, H.B.

    1994-06-01

    Very high cobalt catalysts have been prepared on steamed and acid-washed y zeolite. These catalysts are very active. Some of them have proven to be very stable. Additionally, if provisions are made to control the temperature build up on the catalyst bed, low methane selectivities result. Additional work is indicated, for instance, perhaps even higher activity catalysts can result from higher levels of cobalt or changes in the catalyst preparation procedure. Since the issue of whether small amounts of ruthenium can promote the catalyst is not completely resolved, catalyst preparation procedure experiments should continue with ruthenium vs. ruthenium-free catalysts. For instance, different methods of impregnation and/or activation should be investigated.

  12. SY 17-1 DYNAMIC REGULATION OF REDOX REGULATING FACTOR APE1/REF-1 ON THE OXIDATIVE STRESS AND VASCULAR INFLAMMATION.

    PubMed

    Jeon, Byeong Hwa

    2016-09-01

    Apurinic/apyrimidinic endonuclease 1/redox factor-1 (APE1/Ref-1) is a multifunctional protein that plays a central role in the cellular response to DNA damage and redox regulation against oxidative stress. APE1/Ref-1 is essential for cellular survival and embryonic lethal in knockout mouse models. Heterozygous APE1/Ref-1 mice showed impaired endothelium-dependent vasorelaxation, reduced vascular NO levels, and are hypertensive. APE1/Ref-1 reduces intracellular reactive oxygen species production by negatively regulating the activity of the NADPH oxidase. APE1/Ref-1 is predominantly localized in the nucleus; however, its subcellular localization is dynamically regulated. Recently, it was shown that APE1/Ref-1 is secreted in response to hyperacetylation at specific lysine residues. We investigated the functions of extracellular APE1/Ref-1 with respect to leading anti-inflammatory signaling in TNF-α-stimulated endothelial cells in response to acetylation. Trichostatin A (TSA), an inhibitor of histone deacetylase, considerably suppressed vascular cell adhesion molecule-1 (VCAM-1) in TNF-α-stimulated endothelial cells. During TSA-mediated acetylation in culture, a time-dependent increase in secreted APE1/Ref-1 was confirmed. Recombinant human APE1/Ref-1 with reducing activity induced a conformational change in TNFR1 by thiol-disulfide exchange. Following treatment with the neutralizing anti-APE1/Ref-1 antibody, inflammatory signals via the binding of TNF-α to TNFR1 were remarkably recovered. Furthermore, rhAPE1/Ref-1 inhibited IL-1β-induced VCAM-1 expression in endothelial cells, and it inhibited iNOS or COX-2 expression in lipopolysaccharide-stimulated RAW 264.7 macrophage cells. These results strongly indicate that anti-inflammatory effects of secreted APE1/Ref-1 and its property of secreted APE1/Ref-1 may be useful as a therapeutic biomolecule in cardiovascular disease. PMID:27643268

  13. Biopolymer production from wastewater sludges for encapsulation and other applications. Final report

    SciTech Connect

    Baker, F.R.

    1988-01-01

    The main purpose of the research is to determine the feasibility of producing biopolymers from waste sludges by growing suitable microorganisms on the sludge hydrolysate. Primary and secondary municipal sludges and a bakery waste sludge were used in the study. Enzymatic hydrolysis was found to be ineffective in producing a substrate suitable for the growth of Aureobasidium pullulans (A. pullulans). High pressure acid hydrolysis of municipal sludge tended to yield a better substrate in terms of pullulan yield than low pressure acid hydrolysis. Acid hydrolysis of the bakery waste sludge resulted in a fermentation medium that was highly conductive to the growth of A. pullulans and pullulan elaboration. The feasibility of employing pullulan as a carrier material in the controlled-release of pesticides on stored grains was examined by determining the effect of pullulan-chlorpyrifos-methyl encapsulates on the mortality rate of flour beetles (Tribolilum castaneum).

  14. Paul's gospel and the rhetoric of apostolic rejection: A study of Galatians 1:15--17, 1 Corinthians 15:8, F. C. Baur, and the origins of Paul's Gentile mission

    NASA Astrophysics Data System (ADS)

    Mitchell, Matthew Wesley

    This dissertation proposes a new understanding of Paul's Gentile mission and its relationship to his so-called "conversion." This dissertation examines the origins of Paul's mission to the Gentiles, and locates it in his claims to have been personally commissioned to undertake such a mission by Jesus. Specifically, I argue that it is the rejection of Paul's claim to be an apostle, a claim founded upon his "conversion" experience, that precipitates his mission to the Gentiles. In arguing this view, I draw upon Ferdinand Christian Baur's nineteenth century theories concerning both the unreliability of Acts as a historical source, and his proposal of a clear division between Paul and the other apostles. In establishing the methodological and theoretical framework of the dissertation, I discuss the "New Perspective on Paul" that has dominated New Testament scholarship over the past thirty years. My study is also informed methodologically by the growing interest in rhetorical criticism among biblical scholars, although the emphasis of this dissertation bears more of a resemblance to the approach of the New Rhetoric than the categories of classical, Greco-Roman rhetoric. The textual component of this work falls into two stages. The first contains a full examination of Paul's "conversion passages" in Galatians 1:15--17 and 1 Corinthians 15:8, attempting to situate these seemingly unusual self-descriptions in their cultural contexts. The second involves an examination of F. C. Baur's presentation of Paul, and the reception of Baur's views among biblical scholars throughout the years following his scholarly activity. This dissertation makes two claims, each of which can stand on its own as an important contribution to scholarship. My first claim is that components of Baur's work support my proposal concerning Paul's Gentile mission and his experience of apostolic rejection, and that this proposal has much to commend it as an explanation of a perennial scholarly puzzle. My second claim is methodological, as I demonstrate that scholarly writings about Paul and his modern interpreters are themselves exercises in argumentation, and thus are not to be accepted uncritically, or without close attention to the rhetorical practices they utilize.

  15. Production of novel antistreptococcal liamocins by fermentation of agricultural biomass

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Liamocins are unique heavier-than-water “oils” produced by certain strains of the fungus Aureobasidium pullulans. Liamocins have antibacterial activity with specificity for Streptococcus sp. Previous studies reported that liamocin yields were highest from strains of A. pullulans belonging to phyloge...

  16. Earthworms modify microbial community structure and accelerate maize stover decomposition during vermicomposting.

    PubMed

    Chen, Yuxiang; Zhang, Yufen; Zhang, Quanguo; Xu, Lixin; Li, Ran; Luo, Xiaopei; Zhang, Xin; Tong, Jin

    2015-11-01

    In the present study, maize stover was vermicomposted with the epigeic earthworm Eisenia fetida. The results showed that, during vermicomposting process, the earthworms promoted decomposition of maize stover. Analysis of microbial communities of the vermicompost by high-throughput pyrosequencing showed more complex bacterial community structure in the substrate treated by the earthworms than that in the control group. The dominant microbial genera in the treatment with the earthworms were Pseudoxanthomonas, Pseudomonas, Arthrobacter, Streptomyces, Cryptococcus, Guehomyces, and Mucor. Compared to the control group, the relative abundance of lignocellulose degradation microorganisms increased. The results indicated that the earthworms modified the structure of microbial communities during vermicomposting process, activated the growth of lignocellulose degradation microorganisms, and triggered the lignocellulose decomposition. PMID:26139410

  17. Earthworms modify microbial community structure and accelerate maize stover decomposition during vermicomposting.

    PubMed

    Chen, Yuxiang; Zhang, Yufen; Zhang, Quanguo; Xu, Lixin; Li, Ran; Luo, Xiaopei; Zhang, Xin; Tong, Jin

    2015-11-01

    In the present study, maize stover was vermicomposted with the epigeic earthworm Eisenia fetida. The results showed that, during vermicomposting process, the earthworms promoted decomposition of maize stover. Analysis of microbial communities of the vermicompost by high-throughput pyrosequencing showed more complex bacterial community structure in the substrate treated by the earthworms than that in the control group. The dominant microbial genera in the treatment with the earthworms were Pseudoxanthomonas, Pseudomonas, Arthrobacter, Streptomyces, Cryptococcus, Guehomyces, and Mucor. Compared to the control group, the relative abundance of lignocellulose degradation microorganisms increased. The results indicated that the earthworms modified the structure of microbial communities during vermicomposting process, activated the growth of lignocellulose degradation microorganisms, and triggered the lignocellulose decomposition.

  18. Influence of abiotic variables on culturable yeast diversity in two distinct Alpine glaciers.

    PubMed

    Turchetti, Benedetta; Goretti, Marta; Branda, Eva; Diolaiuti, Guglielmina; D'Agata, Carlo; Smiraglia, Claudio; Onofri, Andrea; Buzzini, Pietro

    2013-11-01

    The influence of some abiotic variables (pH, dry weight, organic carbon, nitrogen and phosphorous) on culturable yeast diversity in two distinct, but adjacent Alpine glaciers (Glacier du Géant, France, and Miage Glacier, Italy) was investigated. In all, 682 yeast strains were isolated and identified by D1/D2 and ITS sequencing as belonging to species of the genera Aureobasidium, Candida, Bulleromyces, Cryptococcus, Cystofilobasidium, Dioszegia, Guehomyces, Holtermanniella, Leucosporidiella, Mrakia, Mrakiella, Rhodotorula, Sporidiobolus, Sporobolomyces and Udenyomyces. Overall, the most represented genera were Cryptococcus (55% of isolates), Rhodotorula (17%) and Mrakia (10%). About 10% of strains, presumably belonging to new species (yet to be described), were preliminarily identified at the genus level. Principal component analysis (PCA) revealed that organic carbon, nitrogen and phosphorous are apparently mostly related to culturable yeast abundance and diversity. In this context, the hypothesis that the frequency of isolation of certain species may be correlated with some organic nutrients (with special emphasis for phosphorous) is discussed.

  19. Paper-based microfluidics with an erodible polymeric bridge giving controlled release and timed flow shutoff.

    PubMed

    Jahanshahi-Anbuhi, Sana; Henry, Aleah; Leung, Vincent; Sicard, Clémence; Pennings, Kevin; Pelton, Robert; Brennan, John D; Filipe, Carlos D M

    2014-01-01

    Water soluble pullulan films were formatted into paper-based microfluidic devices, serving as a controlled time shutoff valve. The utility of the valve was demonstrated by a one-step, fully automatic implementation of a complex pesticide assay requiring timed, sequential exposure of an immobilized enzyme layer to separate liquid streams. Pullulan film dissolution and the capillary wicking of aqueous solutions through the device were measured and modeled providing valve design criteria. The films dissolve mainly by surface erosion, meaning the film thickness mainly controls the shutoff time. This method can also provide time-dependent sequential release of reagents without compromising the simplicity and low cost of paper-based devices.

  20. Interactions between yeasts, fungicides and apple fruit russeting.

    PubMed

    Gildemacher, Peter; Heijne, Bart; Silvestri, Massimiliano; Houbraken, Jos; Hoekstra, Ellen; Theelen, Bart; Boekhout, Teun

    2006-12-01

    The effect of inoculations with yeasts occurring on apple surfaces and fungicide treatments on the russeting of Elstar apples was studied. Captan, dithianon and a water treatment were implemented to study the interaction between the fungicides, the inoculated yeast species and Aureobasidium pullulans, and the development of russet. All yeast inoculations aggravated russet, but Rhodotorula glutinis, Sporidiobolus pararoseus and A. pullulans did so to a greater extent than the other species. Both captan and dithianon significantly reduced russeting. Denaturing gradient gel electrophoresis analysis showed that inoculations with R. glutinis and S. pararoseus seemed to suppress other yeast species present on the apple surface.

  1. Pulsed Laser Processing of Functionalized Polysaccharides for Controlled Release Drug Delivery Systems

    NASA Astrophysics Data System (ADS)

    Cristescu, R.; Popescu, C.; Popescu, A. C.; Socol, G.; Mihailescu, I.; Caraene, G.; Albulescu, R.; Buruiana, T.; Chrisey, D.

    We report on the deposition of triacetate-pullulan polysaccharide thin films on drug pellets (diclofenac sodium) by matrix assisted pulsed laser evaporation method. The radiation generated by a pulsed excimer KrF* laser source (λ = 248 nm, τ = 20 ns) operated at 2 Hz repetition rate was used for ice targets evaporation. The timed - controlled drug delivery was proved by spectroscopic in vitro studies and in vivo anti-inflammatory investigations on rabbits. We showed that the coating of drug pellets with triacetate-pullulan thin films resulted in the delayed delivery of the drug for up to 30 min.

  2. The fungal flora of loganberries in relation to storage and spoilage.

    PubMed

    Davis, R P; Dennis, C

    1977-03-01

    Botrytis cinerea, Cladosporium spp., Penicillium spp., Aureobasidium pullulans, Mucor mucedo, Phoma state of Didymella applanata, Cryptococcus laurentii var. laurentii, C. albidus var. albidus, Candida sake and Sporobolomyces roseus were consistently present on freshly harvested loganberries. Botrytis cinerea was the main spoilage organism with Mucor mucedo of minor importance. The importance of benomyl-resistant strains of B. cinerea is discussed.

  3. Antimicrobial substances from aspen tissue grown in vitro.

    PubMed

    MATHES, M C

    1963-06-01

    Isolated aspen tissue, when grown in vitro for 3 weeks on agar medium, yielded antimicrobial substances which produced inhibitory zones when the culture plates were inoculated with Fusarium roseum, Saccharomyces cervisiae, Bacillus subtilis, Bacillus cereus, Penicillium, roqueforti, Torula utilis, Sarcina lutea, Flavobacterium aquatile, Pullularia pullulans, and Staphylococcus aureus. PMID:13933593

  4. Hydration and energy dissipation measurements of biomolecules on a piezoelectric quartz oscillator by admittance analyses.

    PubMed

    Ozeki, Tomomitsu; Morita, Mizuki; Yoshimine, Hiroshi; Furusawa, Hiroyuki; Okahata, Yoshio

    2007-01-01

    By using a 27-MHz piezoelectric quartz oscillator connected with a vector network analyzer, we obtained resonance frequency decreases (-DeltaFwater) and energy dissipation increases (DeltaDwater) during binding of biotinylated bovine serum albumin, biotinylated ssDNA, biotinylated dsDNA, and biotinylated pullulan to a NeutrAvidin-immobilized 27-MHz quartz crystal microbalance (QCM) plate in aqueous solution, as well as in the wet air phase (98% humidity, -DeltaFwet and DeltaDwet) and in the dry air phase (-DeltaFair and DeltaDair). -DeltaFwater indicates the total mass of the molecule, bound water, and vibrated water in aqueous solutions. -DeltaFwet indicates the total mass of the molecule and bound water. -DeltaFair simply shows the real mass of the molecule on the QCM. In terms of results, (-DeltaFwet)/(-DeltaFair) values indicated the bound water ratios per unit biomolecular mass were on the order of pullulan (2.1-2.2) > DNAs = proteins (1.4-1.6) > polystyrene (1.0). The (-DeltaFwater)/(-DeltaFair) values indicated the hydrodynamic water (bound and vibrated water) ratios per unit biomolecular mass were on the order of dsDNA (6.5) > ssDNA = pullulan (3.5-4.4) > proteins (2.4-2.5) > polystyrene (1.0). Energy dissipation parameters per unit mass in water (DeltaDwater/(-DeltaFair)) were on the order of pullulan > dsDNA > ssDNA > proteins > polystyrene. Energy dissipation in the wet and dry air phases (DeltaDwet and DeltaDair) were negligibly small, which indicates even these biomolecules act as elastic membranes in the air phase (without aqueous solution). We obtained a good linear relationship between [(-DeltaFwater)/(-DeltaFair) - 1], which is indicative of hydration and DeltaDwater/(-DeltaFair) of proteins. The aforementioned values suggest that the energy dissipation of proteins was mainly caused by hydration and that proteins themselves are elastic molecules without energy dissipation in aqueous solutions. On the contrary, plots in cases of denatured proteins

  5. Identification of medically relevant species of arthroconidial yeasts by use of matrix-assisted laser desorption ionization-time of flight mass spectrometry.

    PubMed

    Kolecka, Anna; Khayhan, Kantarawee; Groenewald, Marizeth; Theelen, Bart; Arabatzis, Michael; Velegraki, Aristea; Kostrzewa, Markus; Mares, Mihai; Taj-Aldeen, Saad J; Boekhout, Teun

    2013-08-01

    Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) was used for an extensive identification study of arthroconidial yeasts, using 85 reference strains from the CBS-KNAW yeast collection and 134 clinical isolates collected from medical centers in Qatar, Greece, and Romania. The test set included 72 strains of ascomycetous yeasts (Galactomyces, Geotrichum, Saprochaete, and Magnusiomyces spp.) and 147 strains of basidiomycetous yeasts (Trichosporon and Guehomyces spp.). With minimal preparation time, MALDI-TOF MS proved to be an excellent diagnostic tool that provided reliable identification of most (98%) of the tested strains to the species level, with good discriminatory power. The majority of strains were correctly identified at the species level with good scores (>2.0) and seven of the tested strains with log score values between 1.7 and 2.0. The MALDI-TOF MS results obtained were consistent with validated internal transcribed spacer (ITS) and/or large subunit (LSU) ribosomal DNA sequencing results. Expanding the mass spectrum database by increasing the number of reference strains for closely related species, including those of nonclinical origin, should enhance the usefulness of MALDI-TOF MS-based diagnostic analysis of these arthroconidial fungi in medical and other laboratories.

  6. Methylesterase behaviour is related to polysaccharide organisation in model systems mimicking cell walls.

    PubMed

    Bonnin, Estelle; Mutic, Jelena; Nikolic, Jasna; Burr, Sally; Robert, Paul; Crépeau, Marie-Jeanne

    2015-06-25

    Pectin gels and pectin-cellulose binary gels were used as cell wall-mimicking systems to investigate the diffusion ability of a fungal pectin methylesterase. Increasing content of cellulose in the gel appears to result: (i) in longer demethylated blocks thus favouring AaPME processivity, and (ii) in accelerated enzyme kinetics. To better understand this unexpected behaviour, a method was set up to investigate the gel porosity as a function of the cellulose content by following the passive diffusion of three pullulans having different hydrodynamic volumes. Like the enzyme, the pullulans diffused more efficiently in the gels containing the highest proportions of cellulose. Altogether, these results suggest that the gel settled differently during formation according to the respective proportions of the two polysaccharides. With cellulose present, a fraction of pectin would form close interactions with the microfibrils resulting in a larger volume accessible to diffusing molecules. This volume would be related to the cellulose concentration. PMID:25839794

  7. Characterization of Starch-Debranching Enzymes in Pea Embryos1

    PubMed Central

    Zhu, Zhi-Ping; Hylton, Christopher M.; Rössner, Ute; Smith, Alison M.

    1998-01-01

    Two distinct types of debranching enzymes have been identified in developing pea (Pisum sativum L.) embryos using native gel analysis and tests of substrate preference on purified or partially purified activities. An isoamylase-like activity capable of hydrolyzing amylopectin and glycogen but not pullulan is present throughout development and is largely or entirely confined to the plastid. Activities capable of hydrolyzing pullulan are present both inside and outside of the plastid, and extraplastidial activity increases relative to the plastidial activity during development. Both types of debranching enzyme are also present in germinating embryos. We argue that debranching enzymes are likely to have a role in starch metabolism in the plastid of the developing embryo and in starch degradation during germination. PMID:9765544

  8. Yeasts from native Brazilian Cerrado plants: Occurrence, diversity and use in the biocontrol of citrus green mould.

    PubMed

    Sperandio, Eugenio Miranda; do Vale, Helson Mario Martins; Moreira, Geisianny Augusta Monteiro

    2015-11-01

    Yeasts are some of the most important postharvest biocontrol agents. Postharvest oranges frequently deteriorate due to green mould (Penicillium digitatum), which causes significant losses. The aims of this study were to determine the composition and diversity of yeasts on plants of the Brazilian Cerrado and to explore their potential for inhibiting citrus green mould. Leaves and fruit of Byrsonima crassifolia and Eugenia dysenterica were collected from Cerrado conservation areas, and thirty-five yeasts were isolated and identified by sequencing the D1-D2 domain of the rDNA large subunit (26S). The isolates represented the Aureobasidium, Meyerozyma, Candida, and Pichia genera. Three isolates identified as Aureobasidium pullulans exhibited potential for the control of P. digitatum in both in vitro and in vivo tests; these isolates reduced the incidence of disease and increased the storage time of fruit. Aureobasidium. pullulans has immense potential for the biological control of filamentous fungi.

  9. Superficial Phaeohyphomycosis Caused by Aureobasidium melanogenum Mimicking Tinea Nigra in an Immunocompetent Patient and Review of Published Reports.

    PubMed

    Chen, Wan-Ting; Tu, Mei-Eng; Sun, Pei-Lun

    2016-08-01

    Aureobasidium pullulans is a ubiquitous black yeast-like fungus belonging to order Dothideales. It was regarded as a contaminant, but is now considered a pathogen causing a wide range of human infections. We report a case of superficial phaeohyphomycosis in an immunocompetent patient with clinical presentations mimicking tinea nigra. On microscopic examination of lesion scales, multiple thick-walled, pigmented oval spores with septa were noted. A fungus with black mucoid colonies was repeatedly isolated from the lesions during the treatment course. This fungus was identified as A. melanogenum on the basis of morphological characteristics and subsequently confirmed by sequencing internal transcribed spacers of ribosomal DNA. The clinical presentations and microscopic findings of lesion scales were considerably similar to those of tinea nigra. However, fungal culturing proved that the causative pathogen was A. melanogenum rather than Hortaea werneckii. The patient might have acquired this infection during gardening activities. We also reviewed reported cases of cutaneous A. pullulans infection. PMID:26883514

  10. Childhood hypersensitivity pneumonitis associated with fungal contamination of indoor hydroponics.

    PubMed

    Engelhart, Steffen; Rietschel, Ernst; Exner, Martin; Lange, Lars

    2009-01-01

    Childhood hypersensitivity pneumonitis (HP) is often associated with exposure to antigens in the home environment. We describe a case of HP associated with indoor hydroponics in a 14-year-old girl. Water samples from hydroponics revealed Aureobasidium pullulans as the dominant fungal micro-organism (10(4)CFU/ml). The diagnosis is supported by the existence of serum precipitating antibodies against A. pullulans, lymphocytic alveolitis on bronchoalveolar lavage (BAL) fluid, a corresponding reaction on a lung biopsy, and the sustained absence of clinical symptoms following the removal of hydroponics from the home. We conclude that hydroponics should be considered as potential sources of fungal contaminants when checking for indoor health complaints.

  11. Molecular entanglement and electrospinnability of biopolymers.

    PubMed

    Kong, Lingyan; Ziegler, Gregory R

    2014-01-01

    Electrospinning is a fascinating technique to fabricate micro- to nano-scale fibers from a wide variety of materials. For biopolymers, molecular entanglement of the constituent polymers in the spinning dope was found to be an essential prerequisite for successful electrospinning. Rheology is a powerful tool to probe the molecular conformation and interaction of biopolymers. In this report, we demonstrate the protocol for utilizing rheology to evaluate the electrospinnability of two biopolymers, starch and pullulan, from their dimethyl sulfoxide (DMSO)/water dispersions. Well-formed starch and pullulan fibers with average diameters in the submicron to micron range were obtained. Electrospinnability was evaluated by visual and microscopic observation of the fibers formed. By correlating the rheological properties of the dispersions to their electrospinnability, we demonstrate that molecular conformation, molecular entanglement, and shear viscosity all affect electrospinning. Rheology is not only useful in solvent system selection and process optimization, but also in understanding the mechanism of fiber formation on a molecular level.

  12. Yeasts from native Brazilian Cerrado plants: Occurrence, diversity and use in the biocontrol of citrus green mould.

    PubMed

    Sperandio, Eugenio Miranda; do Vale, Helson Mario Martins; Moreira, Geisianny Augusta Monteiro

    2015-11-01

    Yeasts are some of the most important postharvest biocontrol agents. Postharvest oranges frequently deteriorate due to green mould (Penicillium digitatum), which causes significant losses. The aims of this study were to determine the composition and diversity of yeasts on plants of the Brazilian Cerrado and to explore their potential for inhibiting citrus green mould. Leaves and fruit of Byrsonima crassifolia and Eugenia dysenterica were collected from Cerrado conservation areas, and thirty-five yeasts were isolated and identified by sequencing the D1-D2 domain of the rDNA large subunit (26S). The isolates represented the Aureobasidium, Meyerozyma, Candida, and Pichia genera. Three isolates identified as Aureobasidium pullulans exhibited potential for the control of P. digitatum in both in vitro and in vivo tests; these isolates reduced the incidence of disease and increased the storage time of fruit. Aureobasidium. pullulans has immense potential for the biological control of filamentous fungi. PMID:26466874

  13. Extracellular Proteinases of Yeasts and Yeastlike Fungi1

    PubMed Central

    Ahearn, D. G.; Meyers, S. P.; Nichols, R. A.

    1968-01-01

    Approximately 800 yeasts and other fungi, representing over 70 species, were tested for extracellular caseinolysis. Isolates of a variety of genera, including Aureobasidium, Cephalosporium, Endomycopsis, Kluyveromyces, and numerous sporobolomycetes, demonstrated significant proteolytic activity. Caseinolysis was not necessarily correlated with gelatin liquefaction or with albuminolysis. Numerous fungi showed significant proteolysis at 5 C. The most active organisms were isolates of Candida lipolytica, Aureobasidium pullulans, Candida punicea, and species of Cephalosporium. Taxonomic and ecological implications of proteolytic activity are discussed. Images Fig. 1 PMID:5692110

  14. Volatile emissions from an epiphytic fungus are semiochemicals for eusocial wasps.

    PubMed

    Davis, Thomas Seth; Boundy-Mills, Kyria; Landolt, Peter J

    2012-11-01

    Microbes are ubiquitous on plant surfaces. However, interactions between epiphytic microbes and arthropods are rarely considered as a factor that affects arthropod behaviors. Here, volatile emissions from an epiphytic fungus were investigated as semiochemical attractants for two eusocial wasps. The fungus Aureobasidium pullulans was isolated from apples, and the volatile compounds emitted by fungal colonies were quantified. The attractiveness of fungal colonies and fungal volatiles to social wasps (Vespula spp.) were experimentally tested in the field. Three important findings emerged: (1) traps baited with A. pullulans caught 2750 % more wasps on average than unbaited control traps; (2) the major headspace volatiles emitted by A. pullulans were 2-methyl-1-butanol, 3-methyl-1-butanol, and 2-phenylethyl alcohol; and (3) a synthetic blend of fungal volatiles attracted 4,933 % more wasps on average than unbaited controls. Wasps were most attracted to 2-methyl-1-butanol. The primary wasp species attracted to fungal volatiles were the western yellowjacket (Vespula pensylvanica) and the German yellowjacket (V. germanica), and both species externally vectored A. pullulans. This is the first study to link microbial volatile emissions with eusocial wasp behaviors, and these experiments indicate that volatile compounds emitted by an epiphytic fungus can be responsible for wasp attraction. This work implicates epiphytic microbes as important components in the community ecology of some eusocial hymenopterans, and fungal emissions may signal suitable nutrient sources to foraging wasps. Our experiments are suggestive of a potential symbiosis, but additional studies are needed to determine if eusocial wasp-fungal associations are widespread, and whether these associations are incidental, facultative, or obligate.

  15. The role of N1 domain on the activity, stability, substrate specificity and raw starch binding of amylopullulanase of the extreme thermophile Geobacillus thermoleovorans.

    PubMed

    Nisha, M; Satyanarayana, T

    2015-07-01

    In order to understand the role of N1 domain (1-257 aa) in the amylopullulanase (gt-apu) of the extremely thermophilic bacterium Geobacillus thermoleovorans NP33, N1 deletion construct (gt-apuΔN) has been generated and expressed in Escherichia coli. The truncated amylopullulanase (gt-apuΔN) exhibits similar pH and temperature optima like gt-apu, but enhanced thermostability. The gt-apuΔN has greater hydrolytic action and specific activity on pullulan than gt-apu. The k cat (starch and pullulan) and K m (starch) values of gt-apuΔN increased, while K m (pullulan) decreased. The enzyme upon N1 deletion hydrolyzed maltotetraose as the smallest substrate in contrast to maltopentaose of gt-apu. The role of N1 domain of gt-apu in raw starch binding has been confirmed, for the first time, based on deletion and Langmuir-Hinshelwood kinetics. Furthermore, N1 domain appears to exert a negative influence on the thermostability of gt-apu because N1 truncation significantly improves thermostability. PMID:25573470

  16. Strategies for the production of high-content fructo-oligosaccharides through the removal of small saccharides by co-culture or successive fermentation with yeast.

    PubMed

    Nobre, C; Castro, C C; Hantson, A-L; Teixeira, J A; De Weireld, G; Rodrigues, L R

    2016-01-20

    Fructo-oligosaccharides (FOS) obtained by fermentation of sucrose may be purified at large-scale by continuous chromatography (Simulated Moving Bed: SMB). In order to improve the efficiency of the subsequent SMB purification, the optimization of the fermentative broth composition in salts and sugars was investigated. Fermentations conducted at reduced amount of salts, using Aureobasidium pullulans whole cells, yielded 0.63 ± 0.03 g of FOS per gram of initial sucrose. Additionally, a microbial treatment was proposed to reduce the amount of small saccharides in the mixture. Two approaches were evaluated, namely a co-culture of A. pullulans with Saccharomyces cerevisiae; and a two-step fermentation in which FOS were first synthesized by A. pullulans and then the small saccharides were metabolized by S. cerevisiae. Assays were performed in 100mL shaken flasks and further scaled-up to a 3 L working volume bioreactor. Fermentations in two-step were found to be more efficient than the co-culture ones. FOS were obtained with a purity of 81.6 ± 0.8% (w/w), on a dry weight basis, after the second-step fermentation with S. cerevisiae. The sucrose amount was reduced from 13.5 to 5.4% in total sugars, which suggests that FOS from this culture broth will be more efficiently separated by SMB.

  17. Development of a biodegradable iron oxide nanoparticle gel for tumor bed therapy

    NASA Astrophysics Data System (ADS)

    Cunkelman, B. P.; Chen, E. Y.; Petryk, A. A.; Tate, J. A.; Thappa, S. G.; Collier, R. J.; Hoopes, P. J.

    2013-02-01

    Treatments of the post-operative surgical bed have proven appealing as the majority of cancer recurrence following tumor resection occurs at the tumor margin. A novel, biodegradable pullulan-based gel infused with magnetic iron oxide nanoparticles (IONP) is presented here for surgical bed administration followed by hyperthermia therapy via alternating magnetic field (AMF) activation. Pullulan is a water soluble, film-forming starch polymer that degrades at the postoperative wound site to deliver the IONP payload, targeting the remaining cancer cells. Different gel formulations containing various % wt of pullulan were tested for IONP elution. Elution levels and amount of gel degradation were measured by immersing the gel in de-ionized water for one hour then measuring particle concentrations in the supernatant and the mass of the remaining gel formulation. The most promising gel formulations will be tested in a murine model of surgical bed resection to assess in vivo gel dissolution, IONP cell uptake kinetics via histology and TEM analysis, and heating capability of the gel with AMF exposure.

  18. Multiple rare opportunistic and pathogenic fungi in persistent foot skin infection.

    PubMed

    Chan, Giek Far; Sinniah, Sivaranjini; Idris, Tengku Idzzan Nadzirah Tengku; Puad, Mohamad Safwan Ahmad; Abd Rahman, Ahmad Zuhairi

    2013-03-01

    Persistent superficial skin infection caused by multiple fungi is rarely reported. Recently, a number of fungi, both opportunistic and persistent in nature were isolated from the foot skin of a 24-year old male in Malaysia. The fungi were identified as Candida parapsilosis, Rhodotorula mucilaginosa, Phoma spp., Debaryomyces hansenii, Acremonium spp., Aureobasidium pullulans and Aspergillus spp., This is the first report on these opportunistic strains were co-isolated from a healthy individual who suffered from persistent foot skin infection which was diagnosed as athlete's foot for more than 12 years. Among the isolated fungi, C. parapsilosis has been an increasingly common cause of skin infections. R. mucilaginosa and D. hansenii were rarely reported in cases of skin infection. A. pullulans, an emerging fungal pathogen was also being isolated in this case. Interestingly, it was noted that C. parapsilosis, R. mucilaginosa, D. hansenii and A. pullulans are among the common halophiles and this suggests the association of halotolerant fungi in causing persistent superficial skin infection. This discovery will shed light on future research to explore on effective treatment for inhibition of pathogenic halophiles as well as to understand the interaction of multiple fungi in the progress of skin infection.

  19. The role of N1 domain on the activity, stability, substrate specificity and raw starch binding of amylopullulanase of the extreme thermophile Geobacillus thermoleovorans.

    PubMed

    Nisha, M; Satyanarayana, T

    2015-07-01

    In order to understand the role of N1 domain (1-257 aa) in the amylopullulanase (gt-apu) of the extremely thermophilic bacterium Geobacillus thermoleovorans NP33, N1 deletion construct (gt-apuΔN) has been generated and expressed in Escherichia coli. The truncated amylopullulanase (gt-apuΔN) exhibits similar pH and temperature optima like gt-apu, but enhanced thermostability. The gt-apuΔN has greater hydrolytic action and specific activity on pullulan than gt-apu. The k cat (starch and pullulan) and K m (starch) values of gt-apuΔN increased, while K m (pullulan) decreased. The enzyme upon N1 deletion hydrolyzed maltotetraose as the smallest substrate in contrast to maltopentaose of gt-apu. The role of N1 domain of gt-apu in raw starch binding has been confirmed, for the first time, based on deletion and Langmuir-Hinshelwood kinetics. Furthermore, N1 domain appears to exert a negative influence on the thermostability of gt-apu because N1 truncation significantly improves thermostability.

  20. Characterization and multiple applications of a highly thermostable and Ca²⁺-independent amylopullulanase of the extreme thermophile Geobacillus thermoleovorans.

    PubMed

    Nisha, M; Satyanarayana, T

    2014-12-01

    The amylopullulanase of Geobacillus thermoleovorans NP33 (apu105) is Ca(2+)-independent with a molecular mass of 105 kDa and optimum activity at 80 °C and pH 7.0. The apu105 is extremely thermostable with T 1/2 of 7.8 h at 90 °C and hydrolyzes starch, pullulan, and malto-oligosaccharides, but not panose and cyclodextrins. The low K m values of apu105 (starch, pullulan, amylose, and amylopectin) indicates higher affinity of apu105 than others. The action of the enzyme on mixed substrates (starch and pullulan) confirmed the presence of only one active site for cleaving both α-1,4- and α-1,6- glycosidic linkages. The raw starches are efficiently hydrolyzed into glucose, maltose, and malto-oligosaccharides. Two-step starch saccharification involving pretreatment with apu105 followed by glucoamylase enhanced glucose yield. The supplementation of whole wheat dough with apu105 markedly enhanced the loaf volume, shelf-life, and the texture of bread. The enzyme is compatible with detergents and useful in desizing of cotton fabrics.

  1. Control of postharvest pathogens and colonization of the apple surface by antagonistic microorganisms in the field.

    PubMed

    Leibinger, W; Breuker, B; Hahn, M; Mendgen, K

    1997-11-01

    ABSTRACT Selected isolates of Aureobasidium pullulans, Rhodotorula glutinis, and Bacillus subtilis reduced the size and number of lesions on wounded apples caused by the postharvest pathogens Penicillium expansum, Botrytis cinerea, and Pezicula malicorticis. Combinations of the antagonistic microorganisms were applied to apple trees in the field late in the growing season of two consecutive years. The population dynamics of the introduced microorganisms and the incidence of fruit decay were determined. Population sizes of introduced antagonists on apple surfaces increased in the field following application of treatments until harvest. After transfer of the fruit from the field into cold storage, the populations of the introduced antagonists remained higher than in the control treatments. Identification of the applied isolates of A. pullulans and R. glutinis during the experiments was achieved by isolate-specific DNA probes generated from random amplified polymorphic DNA. A combination of two strains of A. pullulans and one strain of R. glutinis suppressed rotting of apple to the same extent as the commonly used fungicide Euparen. Our data demonstrate that the application of antagonistic microorganisms in the field represents a promising alternative to fungicide treatments to control post-harvest diseases of apple.

  2. Preservation of bacteria in natural polymers.

    PubMed

    Krumnow, April A; Sorokulova, Iryna B; Olsen, Eric; Globa, Ludmila; Barbaree, James M; Vodyanoy, Vitaly J

    2009-08-01

    A new inexpensive and simple method for preserving microorganisms has been developed. Natural polymers of acacia gum and pullulan were used to preserve model bacteria Escherichia coli and Bacillus subtilis via immobilization and storage under various conditions. Formulation of E. coli and B. subtilis in acacia gum significantly increased the viability of both cultures during desiccation at 40 degrees C as well as during the storage at various temperatures and relative humidity. In the ranges of temperatures and humidity used in experiments, the high humidity affected the viability of E. coli more than high temperature. Thermodynamic parameters for E. coli thermal degradation were used for quantification of results and characterization of the preservation process. Viability of B. subtilis in acacia gum polymer was not significantly changed during the storage in the temperature and humidity experiments. The number of viable B. subtilis recovered after storage in pullulan, and in PBS under various humidity conditions was 1-2 logs less in comparison with the number of cells before storage. It was found that acacia gum provides better protection than pullulan for both bacteria during the preservation process.

  3. Amphiphilic biopolymers (amphibiopols) as new surfactants for membrane protein solubilization

    PubMed Central

    Duval-Terrié, Caroline; Cosette, Pascal; Molle, Gérard; Muller, Guy; Dé, Emmanuelle

    2003-01-01

    The aim of this study was to develop new surfactants for membrane protein solubilization, from a natural, biodegradable polymer: the polysaccharide pullulan. A set of amphiphilic pullulans (HMCMPs), differing in hydrophobic modification ratio, charge ratio, and the nature of the hydrophobic chains introduced, were synthesized and tested in solubilization experiments with outer membranes of Pseudomonas fluorescens. The membrane proteins were precipitated, and then resolubilized with various HMCMPs. The decyl alkyl chain (C10) was the hydrophobic graft that gave the highest level of solubilization. Decyl alkyl chain-bearing HMCMPs were also able to extract integral membrane proteins from their lipid environment. The best results were obtained with an amphiphilic pullulan bearing 18% decyl groups (18C10). Circular dichroism spectroscopy and membrane reconstitution experiments were used to test the structural and functional integrity of 18C10-solubilized proteins (OmpF from Escherichia coli and bacteriorhodopsin from Halobacterium halobium). Whatever their structure type (α or β), 18C10 did not alter either the structure or the function of the proteins analyzed. Thus, HMCMPs appear to constitute a promising new class of polymeric surfactants for membrane protein studies. PMID:12649425

  4. Atomistic simulation study of surfactant and polymer interactions on the surface of a fenofibrate crystal.

    PubMed

    Zhu, Wusheng; Romanski, Francis S; Meng, Xiangxin; Mitra, Somenath; Tomassone, M Silvina

    2011-04-18

    It is currently of great interest to the pharmaceutical industry to control the size and agglomeration of nano- and micro-particles for the enhancement of drug delivery. Typically, surfactants and polymers are used as additives to interact with and stabilize the growing crystal surface, thus controlling size and agglomeration; however, selection is traditionally done empirically or using heuristics. The objective of this study was to use molecular dynamic simulations to investigate and predict additive interactions, and thus, evaluate the stabilization potential of individual and multiple additives on the surface of the model drug fenofibrate. Non-ionic surfactant Tween 80, anionic surfactant sodium dodecyl sulfate (SDS), and polymers hydroxypropyl methylcellulose (HPMC) and Pullulan were evaluated individually on three distinct crystal surfaces [(001), (010), (100)], as well as in surfactant-polymer combinations. HPMC was determined to have the strongest interaction with the surfaces of the fenofibrate crystal, and therefore, was predicted to be the most effective individual additive. A mixture of HPMC with SDS was determined to be the most effective mixture of additives, and more effective than HPMC alone, indicating a synergistic effect. The predictions of mixed additives indicated a relative order of effectiveness as follows: HPMC-SDS>HPMC-Tween 80>Pullulan-Tween 80>Pullulan-SDS. The simulations were subsequently validated by an anti-solvent crystallization of fenofibrate where it was found that HPMC individually, and a mixture of HPMC-SDS, produced the smallest and most stable crystals, as measured by laser diffraction; this, in combination with measurements of the crystal growth rate in the presence and absence of additives confirmed the results of the simulations.

  5. Different active biomolecules involved in biosynthesis of gold nanoparticles by three fungus species.

    PubMed

    Zhang, Xiaorong; He, Xiaoxiao; Wang, Kemin; Yang, Xiaohai

    2011-04-01

    In this paper, the intracellular gold nanoparticles were biosynthesized using three fungi including Aureobasidium pullulans (A. pullulans), Fusarium sp. and Fusarium oxysporum (F. oxysporum) after immersion the fungal cells in AuCl4- ions solution. UV-vis and FTIR spectrum, and biochemical compositions analysis of Au nano-fungal cells suggested that active biomolecules of reducing sugar of A. pullulans, and proteins in Fusarium sp. and F. oxysporum were tested positive of providing the function of the reduction of AuCI4- ions and the formation of the gold crystals. SDS-PAGE analysis of purified protein from gold nanoparticles synthesized by three fungi showed that three proteins with molecular weight (WM) about 100 kDa, 25 kDa and 19 kDa were in the gold nanoparticles by Fusarium sp. and two proteins with WM about 25 kDa and 19 kDa were in gold nanoparticles of F oxysporum. Further, three purified fungal proteins with WM about 100 kDa, 25 kDa and 19 kDa from gold nanoparticles by Fusarium sp. identified by LC-MS/MS, named plasma membrane ATPase, 3-glucan binding protein and glyceraldehyde-3-phosphate dehydrogenase, respectively. The Au nano-fungal cells ultrathin sections of Fusarium sp. and F. oxysporum showed that the gold nanoparticles mainly produced in intracellular vacuoles of fungal cells. The growth of gold nanoparticles in three fungal cells indicated the reducing sugar led to the gold nanoparticles in spherical morphology and proteins benefited to the gold aggregates.

  6. Functional Characterization of a Newly Identified Group B Streptococcus Pullulanase Eliciting Antibodies Able to Prevent Alpha-Glucans Degradation

    PubMed Central

    Bosello, Mattia; Berti, Francesco; Mariani, Massimo; Telford, John L.; Grandi, Guido; Soriani, Marco

    2008-01-01

    Streptococcal pullulanases have been recently proposed as key components of the metabolic machinery involved in bacterial adaptation to host niches. By sequence analysis of the Group B Streptococcus (GBS) genome we found a novel putative surface exposed protein with pullulanase activity. We named such a protein SAP. The sap gene is highly conserved among GBS strains and homologous genes, such as PulA and SpuA, have been described in other pathogenic streptococci. The SAP protein contains two N-terminal carbohydrate-binding motifs, followed by a catalytic domain and a C-terminal LPXTG cell wall-anchoring domain. In vitro analysis revealed that the recombinant form of SAP is able to degrade α-glucan polysaccharides, such as pullulan, glycogen and starch. Moreover, NMR analysis showed that SAP acts as a type I pullulanase. Studies performed on whole bacteria indicated that the presence of α-glucan polysaccharides in culture medium up-regulated the expression of SAP on bacterial surface as confirmed by FACS analysis and confocal imaging. Deletion of the sap gene resulted in a reduced capacity of bacteria to grow in medium containing pullulan or glycogen, but not glucose or maltose, confirming the pivotal role of SAP in GBS metabolism of α-glucans. As reported for other streptococcal pullulanases, we found specific anti-SAP antibodies in human sera from healthy volunteers. Investigation of the functional role of anti-SAP antibodies revealed that incubation of GBS in the presence of sera from animals immunized with SAP reduced the capacity of the bacterium to degrade pullulan. Of interest, anti-SAP sera, although to a lower extent, also inhibited Group A Streptococcus pullulanase activity. These data open new perspectives on the possibility to use SAP as a potential vaccine component inducing functional cross-reacting antibodies interfering with streptococcal infections. PMID:19023424

  7. Psychrophilic and mesophilic fungi in frozen food products.

    PubMed

    KUEHN, H H; GUNDERSON, M F

    1963-07-01

    The mold flora of certain frozen pastries and chicken pies was investigated. Molds were determined qualitatively or quantitatively, or both, by preparing pour plates of the blended product and incubating the plates at various temperatures. The mesophilic fungal flora developed on plates incubated at 10 and 20 C, whereas psychrophilic fungi were obtained on plates incubated at 0 and 5 C. About 2,000 cultures of fungi, representing about 100 different species, were isolated from various products. Four different brands of blueberry, two brands of cherry pastries, two brands of apple, and one brand of raspberry pastries were examined. In addition, two brands of chicken pies were studied. Blueberry pastries had a much higher total fungal population than the other products, although different brands of blueberry pastries varied considerably. Blueberry pastries had from 347 to 1,586 psychrophilic fungi per g. Cherry pastries had about 70 to 110 psychrophiles per g, and apple pastries had 19 to 92 psychrophiles per g. Chicken pies contained very few psychrophilic fungi, about 15 per g. Aureobasidium pullulans was recovered most frequently. About 90% of the psychrophilic fungi found in blueberry products was A. pullulans. Depending upon the brand of cherry pastry, either Phoma spp. or A. pullulans was the most common fungus present. Apple pastries also displayed brand variation, but were unique in having many mesophilic aspergilli. This genus was generally absent from other products. The Penicillium content of apple pastries was also rather high; 50% of the psychrophilic flora was represented by this genus. The psychrophilic fungal flora of chicken pies was composed primarily of penicillia (50%) and Chrysosporium pannorum (46%). PMID:13927344

  8. Nanogel-quantum dot hybrid nanoparticles for live cell imaging

    SciTech Connect

    Hasegawa, Urara; Nomura, Shin-ichiro M.; Kaul, Sunil C.; Hirano, Takashi; Akiyoshi, Kazunari; E-mail: akiyoshi.org@tmd.ac.jp

    2005-06-17

    We report here a novel carrier of quantum dots (QDs) for intracellular labeling. Monodisperse hybrid nanoparticles (38 nm in diameter) of QDs were prepared by simple mixing with nanogels of cholesterol-bearing pullulan (CHP) modified with amino groups (CHPNH{sub 2}). The CHPNH{sub 2}-QD nanoparticles were effectively internalized into the various human cells examined. The efficiency of cellular uptake was much higher than that of a conventional carrier, cationic liposome. These hybrid nanoparticles could be a promising fluorescent probe for bioimaging.

  9. Polysaccharide based Copolymers as Supramolecular Systems in Biomedical Applications.

    PubMed

    Célia Monteiro de Paula, Regina; Andrade Feitosa, Judith Pessoa; Beserra Paula, Haroldo César

    2015-01-01

    Polysaccharides are natural polymers, obtained from a large variety of sources ranging from fungi to more complex organisms such as birds and whales. Their use for pharmaceutical and biomedical applications has been the subject of numerous researches by the world´s academia. Polysaccharide chemical/physical modifications leading to graft copolymers are discussed in this review, focusing on those nanosystems that are potential candidates for drug delivery applications. Therefore, this review focuses on the biomedical application of polysaccharide based copolymers, particularly as nanocarriers. Copolymer of polysaccharides such as alginate, cellulose, chitosan, dextran, guar, hyaluronic acid, pullulan and starch as drug delivery nanocarriers will be discussed. PMID:26424388

  10. Screening of micro-organisms for decolorization of melanins produced by bluestain fungi.

    PubMed

    Rättö, M; Chatani, M; Ritschkoff, A C; Viikari, L

    2001-03-01

    A total of 17 fungi and four bacteria were screened for their ability to decolorize melanin, using isolated extracellular melanin of the bluestain fungus Aureobasidium pullulans as substrate. On agar media, decolorization was observed by four fungal strains: Bjerkandera adusta VTT-D-99746, Galactomyces geotrichum VTT-D-84228, Trametes hirsuta VTT-D-95443 and Trametes versicolor VTT-D-99747. The four fungi were more efficient on nitrogen-limited medium than on complete medium. The melanin-decolorizing activity of G. geotrichum appeared to be located on the mycelium and could be liberated into the medium enzymatically.

  11. Target-Induced and Equipment-Free DNA Amplification with a Simple Paper Device.

    PubMed

    Liu, Meng; Hui, Christy Y; Zhang, Qiang; Gu, Jimmy; Kannan, Balamurali; Jahanshahi-Anbuhi, Sana; Filipe, Carlos D M; Brennan, John D; Li, Yingfu

    2016-02-18

    We report on a paper device capable of carrying out target-induced rolling circle amplification (RCA) to produce massive DNA amplicons that can be easily visualized. Interestingly, we observed that RCA was more proficient on paper than in solution, which we attribute to a significantly higher localized concentration of immobilized DNA. Furthermore, we have successfully engineered a fully functional paper device for sensitive DNA or microRNA detection via printing of all RCA-enabling molecules within a polymeric sugar film formed from pullulan, which was integrated with the paper device. This encapsulation not only stabilizes the entrapped reagents at room temperature but also enables colorimetric bioassays with minimal steps.

  12. Hydrolysis of xylan by an immobilized xylanase from Aureobasidium pullanans

    SciTech Connect

    Allenza, P.; Scherl, D.S.; Detroy, R.W.; Leathers, T.D.; Scott, C.D.

    1986-01-01

    The beta-(1,4)-linked xylose residues that comprise the backbone of the abundant plant polymer xylan can be released by enzymic hydrolysis. Xylanase, which is produced in exceptionally high levels by the color-variant strain of A. pullulans, was immobilized onto a macroporous ceramic carrier. Despite a low coupling efficiency, it was possible to run the reactor under a wide range of conditions with flow rates of 3-10 bed volumes/minute of 1% soluble xylan with no detectable leaching of enzyme. The size distribution of products and rate of xylan hydrolysis were very similar for the immobilized and soluble enzymes. (Refs. 13).

  13. Pullulanase: Role in Starch Hydrolysis and Potential Industrial Applications

    PubMed Central

    Hii, Siew Ling; Tan, Joo Shun; Ling, Tau Chuan; Ariff, Arbakariya Bin

    2012-01-01

    The use of pullulanase (EC 3.2.1.41) has recently been the subject of increased applications in starch-based industries especially those aimed for glucose production. Pullulanase, an important debranching enzyme, has been widely utilised to hydrolyse the α-1,6 glucosidic linkages in starch, amylopectin, pullulan, and related oligosaccharides, which enables a complete and efficient conversion of the branched polysaccharides into small fermentable sugars during saccharification process. The industrial manufacturing of glucose involves two successive enzymatic steps: liquefaction, carried out after gelatinisation by the action of α-amylase; saccharification, which results in further transformation of maltodextrins into glucose. During saccharification process, pullulanase has been used to increase the final glucose concentration with reduced amount of glucoamylase. Therefore, the reversion reaction that involves resynthesis of saccharides from glucose molecules is prevented. To date, five groups of pullulanase enzymes have been reported, that is, (i) pullulanase type I, (ii) amylopullulanase, (iii) neopullulanase, (iv) isopullulanase, and (v) pullulan hydrolase type III. The current paper extensively reviews each category of pullulanase, properties of pullulanase, merits of applying pullulanase during starch bioprocessing, current genetic engineering works related to pullulanase genes, and possible industrial applications of pullulanase. PMID:22991654

  14. Breathing silicon anodes for durable high-power operations.

    PubMed

    Hwang, Chihyun; Joo, Sehun; Kang, Na-Ri; Lee, Ungju; Kim, Tae-Hee; Jeon, Yuju; Kim, Jieun; Kim, Young-Jin; Kim, Ju-Young; Kwak, Sang-Kyu; Song, Hyun-Kon

    2015-09-23

    Silicon anode materials have been developed to achieve high capacity lithium ion batteries for operating smart phones and driving electric vehicles for longer time. Serious volume expansion induced by lithiation, which is the main drawback of silicon, has been challenged by multi-faceted approaches. Mechanically rigid and stiff polymers (e.g. alginate and carboxymethyl cellulose) were considered as the good choices of binders for silicon because they grab silicon particles in a tight and rigid way so that pulverization and then break-away of the active mass from electric pathways are suppressed. Contrary to the public wisdom, in this work, we demonstrate that electrochemical performances are secured better by letting silicon electrodes breathe in and out lithium ions with volume change rather than by fixing their dimensions. The breathing electrodes were achieved by using a polysaccharide (pullulan), the conformation of which is modulated from chair to boat during elongation. The conformational transition of pullulan was originated from its α glycosidic linkages while the conventional rigid polysaccharide binders have β linkages.

  15. The effect of locust bean gum (LBG)-based edible coatings carrying biocontrol yeasts against Penicillium digitatum and Penicillium italicum causal agents of postharvest decay of mandarin fruit.

    PubMed

    Parafati, Lucia; Vitale, Alessandro; Restuccia, Cristina; Cirvilleri, Gabriella

    2016-09-01

    Strains belonging to Wickerhamomyces anomalus, Metschnikowia pulcherrima and Aureobasidium pullulans species were tested in vitro as biocontrol agents (BCAs) against the post-harvest pathogenic molds Penicillium digitatum and Penicillium italicum. Moreover, studies aimed at screening the antifungal activity of selected yeast strains in vivo conditions against P. digitatum and P. italicum, and investigated the efficacy of a polysaccharidic matrix, locust bean gum (LBG), enriched with the tested BCAs, in controlling postharvest decays in artificially inoculated mandarins. The population dynamics of BCAs on wounds and the magnitude of peroxidase (POD) and superoxide dismutase (SOD) in fruit tissues were also investigated after treatments of mandarins with antagonistic yeasts. W. anomalus BS91, M. pulcherrima MPR3 and A. pullulans PI1 provided excellent control of postharvest decays caused by P. digitatum and P. italicum on mandarins, both when the yeasts were used alone and in combination with LBG, which enhanced the yeast cell viability over time. Finally, the increased activity of POD and lower decrease in SOD activity in response to BCAs application in mandarin fruits confirmed their involvement in the biocontrol mechanism.

  16. Long-Lasting and Efficient Tumor Imaging Using a High Relaxivity Polysaccharide Nanogel Magnetic Resonance Imaging Contrast Agent.

    PubMed

    Chan, Minnie; Lux, Jacques; Nishimura, Tomoki; Akiyoshi, Kazunari; Almutairi, Adah

    2015-09-14

    Clinically approved small-molecule magnetic resonance imaging (MRI) contrast agents are all rapidly cleared from the body and offer weak signal enhancement. To avoid repeated administration of contrast agent and improve signal-to-noise ratios, agents with stronger signal enhancement and better retention in tumors are needed. Therefore, we focused on hydrogels because of their excellent water accessibility and biodegradability. Gadolinium (Gd)-chelating cross-linkers were incorporated into self-assembled pullulan nanogels to both impart magnetic properties and to stabilize this material that has been extensively studied for medical applications. We show that these Gd-chelating pullulan nanogels (Gd-CHPOA) have the highest reported relaxivity for any hydrogel-based particles and accumulate in the 4T1 tumors in mice at high levels 4 h after injection. This combination offers high signal enhancement and lasts up to 7 days to delineate the tumor clearly for longer imaging time scales. Importantly, this long-term accumulation does not cause any damage or toxicity in major organs up to three months after injection. Our work highlights the clinical potential of Gd-CHPOA as a tumor-imaging MRI contrast agent, permitting tumor identification and assessment with a high signal-to-background ratio.

  17. Transport Systems in Halophilic Fungi.

    PubMed

    Plemenitaš, Ana; Konte, Tilen; Gostinčar, Cene; Cimerman, Nina Gunde

    2016-01-01

    Fungi that tolerate very high environmental NaCl concentrations are good model systems to study mechanisms that enable them to endure osmotic and salinity stress. The whole genome sequences of six such fungal species have been analysed: Hortaea werneckii, Wallemia ichthyophaga and four Aureobasidium spp.: A. pullulans, A. subglaciale, A. melanogenum and A. namibiae. These fungi show different levels of halotolerance, with the presence of numerous membrane transport systems uncovered here that are believed to maintain physiological intracellular concentrations of alkali metal cations. Despite some differences, the intracellular cation contents of H. werneckii, A. pullulans and W. ichthyophaga remain low even under extreme extracellular salinities, which suggests that these species have efficient cation transport systems. We speculate that cation transporters prevent intracellular accumulation of Na(+), and thus avoid the toxic effects that such Na(+) accumulation would have, while also maintaining the high K(+)/Na(+) ratio that is required for the full functioning of the cell - another crucial task in high-Na(+) environments. This chapter primarily summarises the cation transport systems of these selected fungi, and it also describes other membrane transporters that might be involved in their mechanisms of halotolerance.

  18. Pectinolytic yeasts from viticultural and enological environments: novel finding of Filobasidium capsuligenum producing pectinases.

    PubMed

    Merín, María Gabriela; Mendoza, Lucía M; Morata de Ambrosini, Vilma Inés

    2014-08-01

    In this study indigenous yeasts associated with wineries, grapes and Malbec fermented must from San Rafael viticulture region (Argentina) were isolated to select pectinolytic strains for their potential use in enology. Pectinolytic yeasts were identified by physiological and molecular methods. Among 78 isolates, only nine were able to produce extracellular pectinases. Six isolated from berry surface were identified as Aureobasidium pullulans and the remaining isolates, recovered from wineries, belonged to Saccharomyces cerevisiae and Filobasidium capsuligenum species. Pectinase production was evaluated under vinification-related conditions: pH 3.5, 12 and 28 °C. A. pullulans U-12 produced the highest pectinolytic activity at low temperature (1.16 U ml(-1) ), while F. capsuligenum strains showed good activity at 12 and 28 °C (0.77 and 1.15 U ml(-1) , respectively) being this study the first report on the capacity of this species to produce pectinases. The pectinolytic activity of F. capsuligenum B-13 showed an optimum at pH 4.5 and two peaks at 20 and 50 °C. The enzyme half-life was 2 h at 40 °C and retained 65% of its activity at 40 °C after 1 h of incubation. This pectinolytic system displayed remarkable activity at pH and temperatures found in vinification, suggesting a potential candidate for applying to wine-making. PMID:23686851

  19. Yeasts and yeast-like organisms associated with fruits and blossoms of different fruit trees.

    PubMed

    Vadkertiová, Renáta; Molnárová, Jana; Vránová, Dana; Sláviková, Elena

    2012-12-01

    Yeasts are common inhabitants of the phyllosphere, but our knowledge of their diversity in various plant organs is still limited. This study focused on the diversity of yeasts and yeast-like organisms associated with matured fruits and fully open blossoms of apple, plum, and pear trees, during 2 consecutive years at 3 localities in southwest Slovakia. The occurrence of yeasts and yeast-like organisms in fruit samples was 2½ times higher and the yeast community more diverse than that in blossom samples. Only 2 species (Aureobasidium pullulans and Metschnikowia pulcherrima) occurred regularly in the blossom samples, whereas Galactomyces candidus, Hanseniaspora guilliermondii, Hanseniaspora uvarum, M. pulcherrima, Pichia kluyveri, Pichia kudriavzevii, and Saccharomyces cerevisiae were the most frequently isolated species from the fruit samples. The ratio of the number of samples where only individual species were present to the number of samples where 2 or more species were found (consortium) was counted. The occurrence of individual species in comparison with consortia was much higher in blossom samples than in fruit samples. In the latter, consortia predominated. Aureobasidium pullulans, M. pulcherrima, and S. cerevisiae, isolated from both the fruits and blossoms, can be considered as resident yeast species of various fruit tree species cultivated in southwest Slovakia localities. PMID:23210991

  20. Remote detection of laser-induced autofluorescence on pure cultures of fungal and bacterial strains and their analysis with multivariate techniques

    NASA Astrophysics Data System (ADS)

    Raimondi, Valentina; Palombi, Lorenzo; Cecchi, Giovanna; Lognoli, David; Trambusti, Massimo; Gomoiu, Ioana

    2007-05-01

    Remotely sensed laser-induced autofluorescence spectra of pure cultures of fungal strains ( Aureobasidium pullulans, Verticillium sp.) and of bacterial strains ( Bacillus sp., Pseudomonas sp.) are presented. The strains were isolated from samples collected in a Roman archaeological site ( Tropaeum Traiani) near Constanta, Romania. The fluorescence spectra were detected in vivo from a distance of 25 m in the outdoor, using a high spectral resolution fluorescence LIDAR featuring a UV laser (XeCl@308 nm) as an excitation source. All the examined strains, except for the A. pullulans, showed fluorescence features such to allow their characterisation by processing data with multivariate techniques. Both Principal Component Analysis and Cluster Analysis were applied to the data set and compared to discriminate between the examined strains. Results demonstrate the feasibility of fluorescence-based detection and characterisation of fungi and bacteria in the outdoor with a high spectral resolution fluorescence LIDAR. In addition, they show that the proposed processing methods offer a means to discriminate between the fluorescence features due to the investigated samples and that of a fluorescence background of a known spectral shape, as that of the culture medium. This can be exploited for the remote fluorescence mapping of heterotrophic organisms on stone surfaces when the latter show a typical broad fluorescence band.

  1. Gadolinium-chelate nanoparticle entrapped human mesenchymal stem cell via photochemical internalization for cancer diagnosis.

    PubMed

    Kim, Kyoung Sub; Park, Wooram; Na, Kun

    2015-01-01

    To improve the gadolinium (Gd) internalization efficiency in stem cells, gadolinium-chelate nanoparticles were prepared from a pullulan derivative (pullulan-deoxycholic acid (DOCA)-diethylene triamine pentaacetic (DTPA)-Gd conjugate; PDDG) and then the PDDG was entrapped into human mesenchymal stem cells (hMSCs) by the photochemical-internalization (PCI) method for cancer diagnosis via the cancer homing property of hMSCs. The internalization efficiency of Gd in hMSCs was significantly increased to 98 ± 4 pg Gd/cell from 32 ± 2 pg Gd/cell via the PCI method. Moreover, the Gd-entrapped hMSCs revealed a low exocytosis ratio of gadolinium-chelate nanoparticles during cell division in vitro and a high cellular labeling efficiency for at least 21 days in vivo. The cancer-targeting and diagnosis effect of the Gd-entrapped hMSCs were confirmed in a small CT26 tumor-bearing mice model. The stem cells detected an early tumor (∼3 mm(3)) within 2 h using 4.7-T MR and optical imaging. The results demonstrated that the PCI-mediated internalization of Gd-incorporated nanoparticles into hMSCs is a promising protocol for efficient cell labeling and tracking.

  2. Recombinant bacterial amylopullulanases: developments and perspectives.

    PubMed

    Nisha, M; Satyanarayana, T

    2013-01-01

    Pullulanases are endo-acting enzymes capable of hydrolyzing α-1, 6-glycosidic linkages in starch, pullulan, amylopectin, and related oligosaccharides, while amylopullulanases are bifunctional enzymes with an active site capable of cleaving both α-1, 4 and α-1, 6 linkages in starch, amylose and other oligosaccharides, and α-1, 6 linkages in pullulan. The amylopullulanases are classified in GH13 and GH57 family enzymes based on the architecture of catalytic domain and number of conserved sequences. The enzymes with two active sites, one for the hydrolysis of α-1, 4- glycosidic bond and the other for α-1, 6-glycosidic bond, are called α-amylase-pullulanases, while amylopullulanases have only one active site for cleaving both α-1, 4- and α-1, 6-glycosidic bonds. The amylopullulanases produced by bacteria find applications in the starch and baking industries as a catalyst for one step starch liquefaction-saccharification for making various sugar syrups, as antistaling agent in bread and as a detergent additive. PMID:23645215

  3. Thermostable amylolytic enzymes from a new Clostridium isolate

    SciTech Connect

    Madi, E.; Antranikian, G.; Ohmiya, K.; Gottschalk, G.

    1987-07-01

    A new Clostridium strain was isolated on starch at 60 degrees C. Starch, pullulan, maltotriose, and maltose induced the synthesis of alpha-amylase and pullulanase, while glucose, ribose, fructose, and lactose did not. The formation of the amylolytic enzymes was dependent on growth and occurred predominantly in the exponential phase. The enzymes were largely cell bound during growth of the organism with 0.5% starch, but an increase of the starch concentration in the growth medium was accompanied by the excretion of alpha-amylase and pullulanase into the culture broth; but also by a decrease of total activity. Alpha-amylase, pullulanase, and alpha-glucosidase were active in a broad temperature range (40 to 85 degrees C) and displayed temperature optima for activity at 60 to 70 degrees C. During incubation with starch under aerobic conditions at 75 degrees C for 2 hours, the activity of both enzymes decreased to only 90 or 80%. The apparent Km values of alpha-amylase, pullulanase, and alpha-glucosidase for their corresponding substrates, starch, pullulan, and maltose were 0.35 mg/ml, 0.63 mg/ml, and 25 mM, respectively. (Refs. 31).

  4. Yeasts associated with nectarines and their potential for biological control of brown rot.

    PubMed

    Janisiewicz, W J; Kurtzman, C P; Buyer, J S

    2010-07-01

    Resident fruit microflora has been the source of biocontrol agents for the control of postharvest decay of fruits and the active ingredient in commercialized biocontrol products. With the exception of grapes and apples, information on the resident microflora of other fruits is only fragmentary, but greater knowledge in this area can be very helpful in developing biocontrol strategies. We characterized the yeast microflora of nectarines ('Croce del Sud') from the early stages of fruit development until harvest. The fruit samples were collected from trees in an unmanaged orchard. The resident fruit microflora was separated from the occasionally deposited microorganisms by discarding initial fruit washings before the final wash, followed by sonication and plating on NYDA medium. The isolated yeasts were identified by BIOLOG and by sequencing the D1/D2 domain of a large subunit of the rRNA gene and, where available, the ITS sequence. BIOLOG identified 19 and the genetic analysis 23 species of yeasts. Although the identification by these two systems was not always the same, the predominant yeasts were Rhodotorula spp., Sporodiobolus spp., Cryptococcus spp., Pichia spp., Candida spp. and yeast-like Aureobasidium pullulans. Several of the taxa appear to represent new species. The preliminary biocontrol tests against brown rot of nectarine fruit caused by Monilinia fructicola indicates significant decay control potential of some of the identified yeast species, namely Cryptococcus magnus, Cryptococcus sp. nov., Sporidiobolus pararoseus, A. pullulans and Rhodotorula sp. nov.

  5. Isolation and characterization of Schwanniomyces alluvius amylolytic enzymes

    SciTech Connect

    Wilson, J.J.; Ingledew, W.M.

    1982-08-01

    The extracellular amylolytic enzymes of Schwanniomyces alluvius were studied to determine future optimization of this yeast for the production of industrial ethanol from starch. Both alpha-amylase and glucoamylase were isolated and purified. Alpha-amylase has an optimum pH of 6.3 and was stable from pH 4.5 to 7.5. The optimum temperature for the enzyme was 40 degrees C, but it was quickly inactivated at temperatures above 40 degrees C. The Km for soluble starch was 0.364 mg/ml. The molecular weight was calculated to be 61,900 + or - 700. Alpha-amylase was capable of releasing glucose from starch, but not from pullulan. Glucoamylase had an optimum pH of 5.0 and was stable from pH 4.0 to more than 8.0. The optimum temperature for the enzyme was 50 degrees C, and although less heat sensitive than alpha-amylase, it was quickly inactivated at 60 degrees C. Km values were 12.67 mg/ml for soluble starch and 0.72 mM for maltose. The molecular weight was calculated to be 155,000 + or - 3,000. Glucoamylase released only glucose from both soluble starch and pullulan. Schwanniomyces alluvius is one of the very few yeasts to possess both alpha-amylase and glucoamylase as well as some fermentative capacity to produce ethanol. (Refs. 9).

  6. Breathing silicon anodes for durable high-power operations.

    PubMed

    Hwang, Chihyun; Joo, Sehun; Kang, Na-Ri; Lee, Ungju; Kim, Tae-Hee; Jeon, Yuju; Kim, Jieun; Kim, Young-Jin; Kim, Ju-Young; Kwak, Sang-Kyu; Song, Hyun-Kon

    2015-01-01

    Silicon anode materials have been developed to achieve high capacity lithium ion batteries for operating smart phones and driving electric vehicles for longer time. Serious volume expansion induced by lithiation, which is the main drawback of silicon, has been challenged by multi-faceted approaches. Mechanically rigid and stiff polymers (e.g. alginate and carboxymethyl cellulose) were considered as the good choices of binders for silicon because they grab silicon particles in a tight and rigid way so that pulverization and then break-away of the active mass from electric pathways are suppressed. Contrary to the public wisdom, in this work, we demonstrate that electrochemical performances are secured better by letting silicon electrodes breathe in and out lithium ions with volume change rather than by fixing their dimensions. The breathing electrodes were achieved by using a polysaccharide (pullulan), the conformation of which is modulated from chair to boat during elongation. The conformational transition of pullulan was originated from its α glycosidic linkages while the conventional rigid polysaccharide binders have β linkages. PMID:26395407

  7. Yeasts and yeast-like organisms associated with fruits and blossoms of different fruit trees.

    PubMed

    Vadkertiová, Renáta; Molnárová, Jana; Vránová, Dana; Sláviková, Elena

    2012-12-01

    Yeasts are common inhabitants of the phyllosphere, but our knowledge of their diversity in various plant organs is still limited. This study focused on the diversity of yeasts and yeast-like organisms associated with matured fruits and fully open blossoms of apple, plum, and pear trees, during 2 consecutive years at 3 localities in southwest Slovakia. The occurrence of yeasts and yeast-like organisms in fruit samples was 2½ times higher and the yeast community more diverse than that in blossom samples. Only 2 species (Aureobasidium pullulans and Metschnikowia pulcherrima) occurred regularly in the blossom samples, whereas Galactomyces candidus, Hanseniaspora guilliermondii, Hanseniaspora uvarum, M. pulcherrima, Pichia kluyveri, Pichia kudriavzevii, and Saccharomyces cerevisiae were the most frequently isolated species from the fruit samples. The ratio of the number of samples where only individual species were present to the number of samples where 2 or more species were found (consortium) was counted. The occurrence of individual species in comparison with consortia was much higher in blossom samples than in fruit samples. In the latter, consortia predominated. Aureobasidium pullulans, M. pulcherrima, and S. cerevisiae, isolated from both the fruits and blossoms, can be considered as resident yeast species of various fruit tree species cultivated in southwest Slovakia localities.

  8. The effect of locust bean gum (LBG)-based edible coatings carrying biocontrol yeasts against Penicillium digitatum and Penicillium italicum causal agents of postharvest decay of mandarin fruit.

    PubMed

    Parafati, Lucia; Vitale, Alessandro; Restuccia, Cristina; Cirvilleri, Gabriella

    2016-09-01

    Strains belonging to Wickerhamomyces anomalus, Metschnikowia pulcherrima and Aureobasidium pullulans species were tested in vitro as biocontrol agents (BCAs) against the post-harvest pathogenic molds Penicillium digitatum and Penicillium italicum. Moreover, studies aimed at screening the antifungal activity of selected yeast strains in vivo conditions against P. digitatum and P. italicum, and investigated the efficacy of a polysaccharidic matrix, locust bean gum (LBG), enriched with the tested BCAs, in controlling postharvest decays in artificially inoculated mandarins. The population dynamics of BCAs on wounds and the magnitude of peroxidase (POD) and superoxide dismutase (SOD) in fruit tissues were also investigated after treatments of mandarins with antagonistic yeasts. W. anomalus BS91, M. pulcherrima MPR3 and A. pullulans PI1 provided excellent control of postharvest decays caused by P. digitatum and P. italicum on mandarins, both when the yeasts were used alone and in combination with LBG, which enhanced the yeast cell viability over time. Finally, the increased activity of POD and lower decrease in SOD activity in response to BCAs application in mandarin fruits confirmed their involvement in the biocontrol mechanism. PMID:27217363

  9. Yeasts colonizing the leaf surfaces.

    PubMed

    Sláviková, Elena; Vadkertiová, Renata; Vránová, Dana

    2007-08-01

    The yeasts were isolated from the leaf surfaces of ten species of trees. The study site was a forest park (Zelezná Studnicka) of the Small Carpathians mountain range. One hundred and thirty seven yeast strains belonging to 13 genera were isolated from 320 samples of leaves and needles. Seventeen yeast species were isolated, but only seven occurred regularly: Aureobasidium pullulans, Cryptococcus laurentii, Pichia anomala, Metschnikowia pulcherrima, Saccharomyces sp., Lachancea thermotolerans, and Rhodotorula glutinis. The remaining species were isolated from the leaves and needles of three or less tree species. A. pullulans, Cr. laurentii, and P. anomala were the most frequently found species and they occurred on leaves and needles of all ten tree species. Saccharomyces sp. occurred in leaf samples collected from eight kinds of trees. M. pulcherrima and L. thermotolerans were found in samples collected from six species of trees. Both these species occurred almost always on the leaves of deciduous trees. Rh. glutinis was the most frequently isolated carotenoids producing species. We have found out that the ascomycetous and basidiomycetous species were present in the leaf samples in approximately equal frequency, contrary to the soil samples taken from this forest park, where the ascomycetous species were found rarely.

  10. Carbohydrate Availability Regulates Virulence Gene Expression in Streptococcus suis

    PubMed Central

    Ferrando, M. Laura; van Baarlen, Peter; Orrù, Germano; Piga, Rosaria; Bongers, Roger S.; Wels, Michiel; De Greeff, Astrid; Smith, Hilde E.; Wells, Jerry M.

    2014-01-01

    Streptococcus suis is a major bacterial pathogen of young pigs causing worldwide economic problems for the pig industry. S. suis is also an emerging pathogen of humans. Colonization of porcine oropharynx by S. suis is considered to be a high risk factor for invasive disease. In the oropharyngeal cavity, where glucose is rapidly absorbed but dietary α-glucans persist, there is a profound effect of carbohydrate availability on the expression of virulence genes. Nineteen predicted or confirmed S. suis virulence genes that promote adhesion to and invasion of epithelial cells were expressed at higher levels when S. suis was supplied with the α-glucan starch/pullulan compared to glucose as the single carbon source. Additionally the production of suilysin, a toxin that damages epithelial cells, was increased more than ten-fold when glucose levels were low and S. suis was growing on pullulan. Based on biochemical, bioinformatics and in vitro and in vivo gene expression studies, we developed a biological model that postulates the effect of carbon catabolite repression on expression of virulence genes in the mucosa, organs and blood. This research increases our understanding of S. suis virulence mechanisms and has important implications for the design of future control strategies including the development of anti-infective strategies by modulating animal feed composition. PMID:24642967

  11. Gene cloning, functional expression and characterisation of a novel type I pullulanase from Paenibacillus barengoltzii and its application in resistant starch production.

    PubMed

    Liu, Jingjing; Liu, Yu; Yan, Feng; Jiang, Zhengqiang; Yang, Shaoqing; Yan, Qiaojuan

    2016-05-01

    A novel pullulanase gene (PbPulA) from Paenibacillus barengoltzii was cloned. PbPulA has an open reading frame of 2028 bp encoding 675 amino acids. It was heterologously expressed in Escherichia coli as an intracellular soluble protein. The recombinant pullulanase (PbPulA) was purified to homogeneity with a molecular mass of about 75 kDa on SDS-PAGE. PbPulA was optimally active at pH 5.5 and 50 °C. It was stable within pH 5.5-10.5. The enzyme exhibited strict substrate specificity towards pullulan, but showed relatively low activity towards amylopectin and no activity towards other tested polysaccharides. The Km and Vmax values of the enzyme on pullulan were 2.94 mg/mL and 280.5 μmol/min/mg, respectively. The addition of PbPulA in gelatinized rice and maize starches significantly increased the resistant starch type 3 (RS3) yields. Final yields from rice and maize starches were 10.82 g/100 g and 11.41 g/100 g, respectively. These properties make PbPulA useful in starch industries.

  12. Occurrence and infection of Cladosporium, Fusarium, Epicoccum and Aureobasidium in withered rotten grapes during post-harvest dehydration.

    PubMed

    Lorenzini, Marilinda; Zapparoli, Giacomo

    2015-11-01

    Fungi like Cladosporium, Fusarium, Epicoccum and Aureobasidium can occur on withered grapes causing spoilage of passito wine. There is little or no information on the pathogenic role of these fungi. This study describes the isolation, incidence and identification of several isolates from different withered rotten grapes. Representative isolates grouped in several phenotypes were identified by phylogenetic analysis of internal transcribed spacer, actin or elongation factor gene sequences. Isolates of Cladosporium and Fusarium were ascribed to different species, of these C. ramotenellum, C. halotolerans and F. graminearum were isolated from Vitis vinifera for the first time. All Epicoccum and Aureobasidium isolates belonged to E. nigrum and A. pullulans, respectively. Random amplified DNA polymorphism analysis showed high level of heterogenicity among Epicoccum and Fusarium isolates. Infection assays were carried out to evaluate infectivity in some strains under different withering conditions. Fusarium spp. strains had similar infectivity, while significant variability was observed among Cladosporium spp. and E. nigrum strains. A. pullulans resulted particularly infective. This study provided insights into the occurrence and infection of these fungi in fruit-drying rooms with important implications towards control management during the withering.

  13. Molecular Entanglement and Electrospinnability of Biopolymers

    PubMed Central

    Kong, Lingyan; Ziegler, Gregory R.

    2014-01-01

    Electrospinning is a fascinating technique to fabricate micro- to nano-scale fibers from a wide variety of materials. For biopolymers, molecular entanglement of the constituent polymers in the spinning dope was found to be an essential prerequisite for successful electrospinning. Rheology is a powerful tool to probe the molecular conformation and interaction of biopolymers. In this report, we demonstrate the protocol for utilizing rheology to evaluate the electrospinnability of two biopolymers, starch and pullulan, from their dimethyl sulfoxide (DMSO)/water dispersions. Well-formed starch and pullulan fibers with average diameters in the submicron to micron range were obtained. Electrospinnability was evaluated by visual and microscopic observation of the fibers formed. By correlating the rheological properties of the dispersions to their electrospinnability, we demonstrate that molecular conformation, molecular entanglement, and shear viscosity all affect electrospinning. Rheology is not only useful in solvent system selection and process optimization, but also in understanding the mechanism of fiber formation on a molecular level. PMID:25226274

  14. Plasmid DNA-entrapped nanoparticles engineered from microemulsion precursors: in vitro and in vivo evaluation.

    PubMed

    Cui, Zhengrong; Mumper, Russell J

    2002-01-01

    Nonviral gene therapy has been a rapidly growing field. However, delivery systems that can provide protection for pDNA and potential targeting are still desired. A novel pDNA-nanoparticle delivery system was developed by entrapping hydrophobized pDNA inside nanoparticles engineered from oil-in-water (O/W) microemulsion precursors. Plasmid DNA was hydrophobized by complexing with cationic surfactants DOTAP and DDAB. Warm O/W microemulsions were prepared at 50-55 degrees C with emulsifying wax, Brij 78, Tween 20, and Tween 80. Nanoparticles were engineered by simply cooling the O/W microemulsions containing the hydrophobized pDNA in the oil phase to room temperature while stirring. The nanoparticles were characterized by particle sizing, zeta-potential, and TEM. Nanoparticles were challenged with serum nucleases to assess pDNA stability. In addition, the nanoparticles were coincubated with simulated biological media to assess their stability. In vitro hepatocyte transfection studies were completed with uncoated nanoparticles or nanoparticles coated with pullulan, a hepatocyte targeting ligand. In vivo biodistribution of the nanoparticles containing I-125 labeled pDNA was monitored 30 min after tail-vein injection to Balb/C mice. Depending on the hydrophobizing lipid agent employed, uniform pDNA-entrapped nanoparticles (100-160 nm in diameter) were engineered within minutes from warm O/W microemulsion precursors. The nanoparticles were negatively charged (-6 to -15 mV) and spherical. An anionic exchange column was used to separate unentrapped pDNA from nanoparticles. Gel permeation chromatography of pDNA-entrapped and serum-digested nanoparticles showed that the incorporation efficiency was approximately 30%. Free 'naked' pDNA was completely digested by serum nucleases while the entrapped pDNA remained intact. Moreover, in vitro transfection studies in Hep G2 cells showed that pullulan-coated nanoparticles resulted in enhanced luciferase expression, compared to both p

  15. Plasmid DNA-entrapped nanoparticles engineered from microemulsion precursors: in vitro and in vivo evaluation.

    PubMed

    Cui, Zhengrong; Mumper, Russell J

    2002-01-01

    Nonviral gene therapy has been a rapidly growing field. However, delivery systems that can provide protection for pDNA and potential targeting are still desired. A novel pDNA-nanoparticle delivery system was developed by entrapping hydrophobized pDNA inside nanoparticles engineered from oil-in-water (O/W) microemulsion precursors. Plasmid DNA was hydrophobized by complexing with cationic surfactants DOTAP and DDAB. Warm O/W microemulsions were prepared at 50-55 degrees C with emulsifying wax, Brij 78, Tween 20, and Tween 80. Nanoparticles were engineered by simply cooling the O/W microemulsions containing the hydrophobized pDNA in the oil phase to room temperature while stirring. The nanoparticles were characterized by particle sizing, zeta-potential, and TEM. Nanoparticles were challenged with serum nucleases to assess pDNA stability. In addition, the nanoparticles were coincubated with simulated biological media to assess their stability. In vitro hepatocyte transfection studies were completed with uncoated nanoparticles or nanoparticles coated with pullulan, a hepatocyte targeting ligand. In vivo biodistribution of the nanoparticles containing I-125 labeled pDNA was monitored 30 min after tail-vein injection to Balb/C mice. Depending on the hydrophobizing lipid agent employed, uniform pDNA-entrapped nanoparticles (100-160 nm in diameter) were engineered within minutes from warm O/W microemulsion precursors. The nanoparticles were negatively charged (-6 to -15 mV) and spherical. An anionic exchange column was used to separate unentrapped pDNA from nanoparticles. Gel permeation chromatography of pDNA-entrapped and serum-digested nanoparticles showed that the incorporation efficiency was approximately 30%. Free 'naked' pDNA was completely digested by serum nucleases while the entrapped pDNA remained intact. Moreover, in vitro transfection studies in Hep G2 cells showed that pullulan-coated nanoparticles resulted in enhanced luciferase expression, compared to both p

  16. Selection of antagonists of postharvest apple parasites: Penicillium expansum and Botrytis cinerea.

    PubMed

    Achbani, E H; Mounir, R; Jaafari, S; Douira, A; Benbouazza; Jijakli, M H

    2005-01-01

    The objectives of this study were to constitute a collection of pathogenic agents of economic importance which cause losses of apple fruits after harvest namely Botrytis cinerea and Penicillium expansum and to select in vivo efficient antagonistic strains able to protect fruits against both pathogens at 5 degrees C (P. expansum) and 25 degrees C (P. expansum & B. cinerea). Twenty strains of P. expansum and ten strains of B. cinerea have been isolated from infected apple fruits. Potential antagonistic micro-organisms (thirty three isolates) belonging to yeast, bacteria and fungi have been isolated from apple surface. Six of them (strains Ach1-1, Ach2-1, Ach2-2 belonging to Aureobasidium pullulans (De Bary) Arnaud, and strains 1112-3, 1113-10 and 1113-5 belonging to Aureobasidium pullulans (de Bary) Am. v. pullulans) showed a high level of protection (more than 80%) at 25 degrees C. once inoculated with P. expansum or B. cinerea for 5 days. The highest level of protection against P. expansum (96%) was observed with the application of Ach 2-1. Six days after inoculation of B. cinerea, strains Ach 2-2 and Ach 2-1 insured 100% and 96% of protection, respectively. At lower temperature (5 degrees C), first symptoms of P. expansum appeared 13 days after its inoculation. Percentages of protection observed after apple treatment with one of the six antagonistic strains were ranged from 78% to 94% 20 days after P. expansum inoculation. Strains labelled Ach showed a protective level higher than 90% against this pathogen, followed by strain 1113-10 (90%), strain 1113-5 (89%) and strain 1112-3 (82%). At 26 days post-inoculation, levels of protection decreased but remained higher than 60% (more than 80% with strain Ach2-2 and strain 1113-5, 75% with strain Ach2-1 and 1113-10, 72% with ach1-1, 61% for the other strains). Strain Ach2-2 and 1113-10 were retained as the best antagonists for the subsequent studies.

  17. Production of Cellulolytic and Hemicellulolytic Enzymes From Aureobasidium pulluans on Solid State Fermentation

    NASA Astrophysics Data System (ADS)

    Leite, Rodrigo Simões Ribeiro; Bocchini, Daniela Alonso; da Silva Martins, Eduardo; Silva, Dênis; Gomes, Eleni; da Silva, Roberto

    This article investigates a strain of the yeast Aureobasidium pullulans for cellulase and hemicellulase production in solid state fermentation. Among the substrates analyzed, the wheat bran culture presented the highest enzymatic production (1.05 U/mL endoglucanase, 1.3 U/mL β-glucosidase, and 5.0 U/mL xylanase). Avicelase activity was not detected. The optimum pH and temperature for xylanase, endoglucanase and β-glucosidase were 5.0 and 50, 4.5 and 60, 4.0 and 75°C, respectively. These enzymes remained stable between a wide range of pH. The β-glucosidase was the most thermostable enzyme remaining 100% active when incubated at 75°C for 1 h.

  18. Formation of isoamylase by Pseudomonas.

    PubMed

    Harada, T; Yokobayashi, K; Misaki, A

    1968-10-01

    We have isolated a Pseudomonas sp. (strain SB15) which produces an isoamylase (EC 3.2.1.9). Highest yields of this enzyme were obtained when the bacterium was grown in shaken culture in a medium containing maltose, dextrin, starch, or isomaltose. Specific carbon and nitrogen sources were required for growth. The most satisfactory medium consisted of 2% maltose, 0.4% sodium glutamate, 0.3% diammonium hydrogen phosphate, and other inorganic salts. The optimal pH for enzyme production was 5 to 6. The enzyme is stable between pH 3 and 6 but is extremely labile above pH 7. It splits amylopectin completely by combined action with beta-amylase but does not attack pullulan. PMID:5684197

  19. Purification of extrachloroplastic. beta. -amylase from leaves of starchless and wild type Arabidopsis

    SciTech Connect

    Somerville, C.; Monroe, J.; Preiss, J. )

    1989-04-01

    Amylase activity in crude leaf extracts from starchless mutants of Arabidopsis thaliana is 5 to 10 fold higher than in the wild type (WT) when plants are grown under a 12 h photoperiod. Visualized on native PAGE, the increased activity is attributed primarily to a previously characterized extrachloroplastic {beta}-(exo)amylase. The {beta}-amylases from phosoglucomutase deficient (starchless) and WT leaves were purified to homogeneity in two steps utilizing polyethylene glycol fractionation, and cyclohexaamylose affinity chromatography. The enzyme from both mutant and WT leaves had negligible activity toward either {beta}-limit dextrin or pullulan. The specific activities of both purified enzymes were similar indicating that the protein is over-expressed in the mutant. Preliminary antibody neutralization experiments suggest that the two {beta}-amylases are not different.

  20. Extracellular Polysaccharides Produced by Yeasts and Yeast-Like Fungi

    NASA Astrophysics Data System (ADS)

    van Bogaert, Inge N. A.; de Maeseneire, Sofie L.; Vandamme, Erick J.

    Several yeasts and yeast-like fungi are known to produce extracellular polysaccharides. Most of these contain D-mannose, either alone or in combination with other sugars or phosphate. A large chemical and structural variability is found between yeast species and even among different strains. The types of polymers that are synthesized can be chemically characterized as mannans, glucans, phosphoman-nans, galactomannans, glucomannans and glucuronoxylomannans. Despite these differences, almost all of the yeast exopolysaccharides display some sort of biological activity. Some of them have already applications in chemistry, pharmacy, cosmetics or as probiotic. Furthermore, some yeast exopolysaccharides, such as pullulan, exhibit specific physico-chemical and rheological properties, making them useful in a wide range of technical applications. A survey is given here of the production, the characteristics and the application potential of currently well studied yeast extracellular polysaccharides.

  1. Sugarbeet as a renewable resource

    SciTech Connect

    Edye, L.A.; Clarke, M.A.

    1995-12-01

    Sugarbeet (Beta vulgaris) is produced annually on the order of 400 million tonnes, in temperate climates. The primary product is sugar (sucrose); other products include feeds (molasses and beet pulp), and raffinose, pectin and arabinan. Recently, production of paper from sugarbeet pulp has begun. A wide range of non-food products is available through microbial and chemical reactions on sugarbeet juices, molasses and sugars. Products of microbial processes (chemical transformations are discussed in the companion presentation on sugarcane) include polymers to use as biodegradable plastics (pullulans, polyhydroxyalkanoates, polylactide) and others for food and non food use (levan, dextran). Basic chemicals, including citric acid and lactic acid, and amino acids, notably lysine, are produced from sugarbeet sources. The production of ethanol, as fuel or as beverage, is well known. Products and processes are outlined, and recent developments are emphasized.

  2. Target-Induced and Equipment-Free DNA Amplification with a Simple Paper Device.

    PubMed

    Liu, Meng; Hui, Christy Y; Zhang, Qiang; Gu, Jimmy; Kannan, Balamurali; Jahanshahi-Anbuhi, Sana; Filipe, Carlos D M; Brennan, John D; Li, Yingfu

    2016-02-18

    We report on a paper device capable of carrying out target-induced rolling circle amplification (RCA) to produce massive DNA amplicons that can be easily visualized. Interestingly, we observed that RCA was more proficient on paper than in solution, which we attribute to a significantly higher localized concentration of immobilized DNA. Furthermore, we have successfully engineered a fully functional paper device for sensitive DNA or microRNA detection via printing of all RCA-enabling molecules within a polymeric sugar film formed from pullulan, which was integrated with the paper device. This encapsulation not only stabilizes the entrapped reagents at room temperature but also enables colorimetric bioassays with minimal steps. PMID:26748431

  3. Fungal Exopolysaccharide: Production, Composition and Applications

    PubMed Central

    Mahapatra, Subhadip; Banerjee, Debdulal

    2013-01-01

    Fungal exopolysaccharides (EPSs) have been recognized as high value biomacromolecules for the last two decades. These products, including pullulan, scleroglucan, and botryosphaeran, have several applications in industries, pharmaceuticals, medicine, foods etc. Although fungal EPSs are highly relevant, to date information concerning fungal biosynthesis is scarce and an extensive search for new fugal species that can produce novel EPSs is still needed. In most cases, the molecular weight variations and sugar compositions of fungal EPSs are dependent to culture medium composition and different physical conditions provided during fermentation. An inclusive and illustrative review on fungal EPS is presented here. The general outline of the present work includes fungal EPS production, their compositions and applications. An emphasis is also given to listing out different fungal strains that can produce EPSs. PMID:24826070

  4. Formation of polyelectrolyte complexes with diethylaminoethyl dextran: charge ratio and molar mass effect.

    PubMed

    Le Cerf, Didier; Pepin, Anne Sophie; Niang, Pape Momar; Cristea, Mariana; Karakasyan-Dia, Carole; Picton, Luc

    2014-11-26

    The formation of polyelectrolyte complexes (PECs) between carboxymethyl pullulan and DEAE Dextran, was investigated, in dilute solution, with emphasis on the effect of charge density (molar ratio or pH) and molar masses. Electrophoretic mobility measurements have evidenced that insoluble PECs (neutral electrophoretic mobility) occurs for charge ratio between 0.6 (excess of polycation) and 1 (stoichiometry usual value) according to the pH. This atypical result is explained by the inaccessibility of some permanent cationic charge when screened by pH dependant cationic ones (due to the Hoffman alkylation). Isothermal titration calorimetry (ITC) indicates an endothermic formation of PEC with a binding constant around 10(5) L mol(-1). Finally asymmetrical flow field flow fractionation coupled on line with static multi angle light scattering (AF4/MALS) evidences soluble PECs with very large average molar masses and size around 100 nm, in agreement with scrambled eggs multi-association between various polyelectrolyte chains.

  5. Development of taste masked fast disintegrating films of levocetirizine dihydrochloride for oral use.

    PubMed

    Mahesh, A; Shastri, Nalini; Sadanandam, M

    2010-01-01

    Fast disintegrating films of levocetirizine dihydrochloride useful for the treatment of acute allergic rhinitis and chronic urticaria have been developed by using the taste masking ability of cyclodextrins. The fast disintegrating films were prepared by solvent casting method. The films contained water-soluble polymers such as Kollicoat IR or pullulan, aspartame and sucralose as sweeteners and pre-gelatinized starch as disintegrant. Levocetirizine dihydrochloride was incorporated into these films by in-situ complex formation with hydroxy propyl beta-cyclodextrin. The optimized films were evaluated for weight variation, film thickness, folding endurance, tackiness, tensile strength, assay, content uniformity, in vitro disintegration and dissolution, in vivo disintegration and taste masking ability by human gustatory sensation test. Results revealed that the organoleptic properties of levocetirizine dihydrochloride were improved by complexation with hydroxy propyl beta-cyclodextrin and the complex could be successfully formulated into a fast disintegrating film. PMID:19863484

  6. Fungal endophytes in germinated seeds of the common bean, Phaseolus vulgaris

    PubMed Central

    Parsa, Soroush; García-Lemos, Adriana M.; Castillo, Katherine; Ortiz, Viviana; López-Lavalle, Luis Augusto Becerra; Braun, Jerome; Vega, Fernando E.

    2016-01-01

    We conducted a survey of fungal endophytes in 582 germinated seeds belonging to 11 Colombian cultivars of the common bean (Phaseolus vulgaris). The survey yielded 394 endophytic isolates belonging to 42 taxa, as identified by sequence analysis of the ribosomal DNA internal transcribed spacer (ITS) region. Aureobasidium pullulans was the dominant endophyte, isolated from 46.7 % of the samples. Also common were Fusarium oxysporum, Xylaria sp., and Cladosporium cladosporioides, but found in only 13.4 %, 11.7 %, and 7.6 % of seedlings, respectively. Endophytic colonization differed significantly among common bean cultivars and seedling parts, with the highest colonization occurring in the first true leaves of the seedlings. PMID:27109374

  7. Formation of polyelectrolyte complexes with diethylaminoethyl dextran: charge ratio and molar mass effect.

    PubMed

    Le Cerf, Didier; Pepin, Anne Sophie; Niang, Pape Momar; Cristea, Mariana; Karakasyan-Dia, Carole; Picton, Luc

    2014-11-26

    The formation of polyelectrolyte complexes (PECs) between carboxymethyl pullulan and DEAE Dextran, was investigated, in dilute solution, with emphasis on the effect of charge density (molar ratio or pH) and molar masses. Electrophoretic mobility measurements have evidenced that insoluble PECs (neutral electrophoretic mobility) occurs for charge ratio between 0.6 (excess of polycation) and 1 (stoichiometry usual value) according to the pH. This atypical result is explained by the inaccessibility of some permanent cationic charge when screened by pH dependant cationic ones (due to the Hoffman alkylation). Isothermal titration calorimetry (ITC) indicates an endothermic formation of PEC with a binding constant around 10(5) L mol(-1). Finally asymmetrical flow field flow fractionation coupled on line with static multi angle light scattering (AF4/MALS) evidences soluble PECs with very large average molar masses and size around 100 nm, in agreement with scrambled eggs multi-association between various polyelectrolyte chains. PMID:25256478

  8. Development of taste masked fast disintegrating films of levocetirizine dihydrochloride for oral use.

    PubMed

    Mahesh, A; Shastri, Nalini; Sadanandam, M

    2010-01-01

    Fast disintegrating films of levocetirizine dihydrochloride useful for the treatment of acute allergic rhinitis and chronic urticaria have been developed by using the taste masking ability of cyclodextrins. The fast disintegrating films were prepared by solvent casting method. The films contained water-soluble polymers such as Kollicoat IR or pullulan, aspartame and sucralose as sweeteners and pre-gelatinized starch as disintegrant. Levocetirizine dihydrochloride was incorporated into these films by in-situ complex formation with hydroxy propyl beta-cyclodextrin. The optimized films were evaluated for weight variation, film thickness, folding endurance, tackiness, tensile strength, assay, content uniformity, in vitro disintegration and dissolution, in vivo disintegration and taste masking ability by human gustatory sensation test. Results revealed that the organoleptic properties of levocetirizine dihydrochloride were improved by complexation with hydroxy propyl beta-cyclodextrin and the complex could be successfully formulated into a fast disintegrating film.

  9. Effect of simulated acid rain on quantity of epiphytic microfungi on Scots pine (Pinus sylvestris L.) needles.

    PubMed

    Ranta, H M

    1990-01-01

    The effects of simulated acid rain on the phyllosphere microflora of pine (Pinus sylvestris L.) were studied experimentally in northern Finland during the summer of 1988. Trees were irrigated with artificial acid rain of pH4 and pH3 (H(2)SO(4) and HNO(3), weight ratio of S:N=2.86:1). Untreated trees and trees irrigated with spring water (pH6) were used as controls. Two sampling heights (0.5m and 2m) were used. The needles were colonized exclusively by epiphytic fungi, mainly Aureobasidium pullulans (de Bary) Arnaud. The lower branches had significantly more epiphytic fungi than the upper branches. Compared to the control trees, the numbers of epiphytic fungi were significantly decreased on the needles of trees irrigated with acid rain. Acid rain affected the number of epiphytic fungi equally at both sampling heights. The species composition of the epiphytic fungi was not affected by the acid treatments.

  10. Effects of sperminated polymers on the pulmonary absorption of insulin.

    PubMed

    Seki, Toshinobu; Fukushi, Nanako; Chono, Sumio; Morimoto, Kazuhiro

    2008-02-11

    Sperminated pullulans (SP) and gelatin (SG) having different numbers of amino groups were prepared as sperminated polymers for absorption enhancement and the absorption-enhancing effects on the pulmonary absorption of insulin in rats and the permeation of FITC-dextran (MW 4400, FD4) through Calu-3 cell monolayers were evaluated. In both in vivo and in vitro experimental systems, SP and SG acted as enhancers and their enhancing effects correlated with the amino group content. The zeta potential of red blood cells treated with SP and SG was positive and the degree was also correlated with the amino group content. SP and SG interact directly with the luminal surface of mucus membranes via an ion-ion interaction and then may induce signals that open tight junctions resulting in intercellular permeation of water soluble drugs.

  11. Pressate from peat dewatering as a substrate for bacterial growth. [Rhizopus arrhizus; Xanthomonas campestris; Aureobasidium

    SciTech Connect

    Mulligan, C.N.; Cooper, D.G.

    1985-07-01

    This study considered the possibility of using water expressed during the drying of fuel-grade peat as a substrate for microbial growth. Highly humified peat pressed for 2.5 min at 1.96 MPa produced water with a chemical oxygen demand of 690 mg/liter. Several biological compounds could be produced by using the organic matter inexpressed peat water as a substrate. These included polymers such as chitosan, contained in the cell wall of Rhizopus arrhizus, and two extracellular polysaccharides, xanthan gum and pullulan, produced by Bacillus subtilis grown in the expressed water. Small additions of nutrients to the peat pressate were necessary to obtain substantial yields of products. The addition of peptone, yeast extract, and glucose improved production of the various compounds. Biological treatment improved the quality of the expressed water to the extent that in an industrial process it could be returned to the environment.

  12. The use of cationic nanogels to deliver proteins to myeloma cells and primary T lymphocytes that poorly express heparan sulfate.

    PubMed

    Watanabe, Kozo; Tsuchiya, Yumiko; Kawaguchi, Yoshinori; Sawada, Shin-ichi; Ayame, Hirohito; Akiyoshi, Kazunari; Tsubata, Takeshi

    2011-09-01

    Fusion proteins containing protein transduction domain (PTD) are widely used for intracellular delivery of exogenous proteins. PTD-mediated delivery requires expression of heparan sulfate on the surface of the target cells. However, some of metastatic tumor cells and primary lymphocytes poorly express heparan sulfate. Here we demonstrate that proteins complexed with nanosize hydrogels formed by cationic cholesteryl group-bearing pullulans (cCHP) are efficiently delivered to myeloma cells and primary CD4(+) T lymphocytes probably by induction of macropinocytosis, although these cells are resistant to PTD-mediated protein delivery as a consequence of poor heparan sulfate expression. The anti-apoptotic protein Bcl-xL delivered by cCHP nanogels efficiently blocked apoptosis of these cells, establishing functional regulation of cells by proteins delivered by cCHP nanogels. Thus, cCHP nanogel is a useful tool to deliver proteins for development of new cancer therapy and immune regulation. PMID:21605901

  13. Therapeutic Trials Based on Combination of Drug Delivery System and Ultrasound

    NASA Astrophysics Data System (ADS)

    Tabata, Yasuhiko

    2005-03-01

    The objective of this study is to show how therapeutically effective a combination of a drug delivery system (DDS) and ultrasound (US) is by introducing data from two related experiments. Fullerene (C60) was chemically modified by polyethylene glycol (PEG) for water-solubilization and tumour targeting. When the PEG-modified C60 was injected intravenously into tumour bearing mice, followed by US irradiation to the tumour site, a synergistic anti-tumour effect was observed. Following the intravenous injection of interferon (IFN) conjugated with pullulan which has an inherent affinity for the liver, and subsequent US irradiation to the liver, the activity level of an IFN-specific enzyme was significantly enhanced compared with that of the conjugate injection alone.

  14. A biotechnological perspective on the application of iron oxide magnetic colloids modified with polysaccharides.

    PubMed

    Dias, A M G C; Hussain, A; Marcos, A S; Roque, A C A

    2011-01-01

    Iron oxide magnetic nanoparticles (MNPs) alone are suitable for a broad spectrum of applications, but the low stability and heterogeneous size distribution in aqueous medium represent major setbacks. These setbacks can however be reduced or diminished through the coating of MNPs with various polymers, especially biopolymers such as polysaccharides. Polysaccharides are biocompatible, non-toxic and renewable; in addition, they possess chemical groups that permit further functionalization of the MNPs. Multifunctional entities can be created through decoration with specific molecules e.g. proteins, peptides, drugs, antibodies, biomimetic ligands, transfection agents, cells, and other ligands. This development opens a whole range of applications for iron oxide nanoparticles. In this review the properties of magnetic structures composed of MNPs and several polysaccharides (Agarose, Alginate, Carrageenan, Chitosan, Dextran, Heparin, Gum Arabic, Pullulan and Starch) will be discussed, in view of their recent and future biomedical and biotechnological applications.

  15. A hybrid hydrogel biomaterial by nanogel engineering: bottom-up design with nanogel and liposome building blocks to develop a multidrug delivery system.

    PubMed

    Sekine, Yurina; Moritani, Yuki; Ikeda-Fukazawa, Tomoko; Sasaki, Yoshihiro; Akiyoshi, Kazunari

    2012-11-01

    New hybrid poly(ethylene glycol) (PEG) hydrogels crosslinked with both nanogels and nanogel-coated liposome complexes are obtained by Michael addition of the acryloyl group of a cholesterol-bearing pullulan (CHP) nanogel to the thiol group of pentaerythritol tetra(mercaptoethyl) polyoxyethylene. The nanogel-coated liposome complex is stably retained after gelation and the complexes are well dispersed in the hybrid gel. Microrheological measurements show that the strength and gelation time of the hybrid hydrogel can be controlled by changing the liposome:nanogel ratio. The hydrogel is gradually degraded by hydrolysis under physiological conditions. In this process, the nanogel is released first, followed by the nanogel-coated liposomes. Hybrid hydrogels that can incorporate various molecules into the nanogel and liposomes, and release them in a two-step controllable manner, represent a new functional scaffold capable of delivering multiple drugs, proteins or DNA.

  16. Anti-inflammatory effect of water-soluble complex of 1'-acetoxychavicol acetate with highly branched β-1,3-glucan on contact dermatitis.

    PubMed

    Li, Jiawei; Aizawa, Yui; Hiramoto, Keiichi; Kasahara, Emiko; Tsuruta, Daisuke; Suzuki, Toshio; Ikeda, Atsushi; Azuma, Hideki; Nagasaki, Takeshi

    2015-02-01

    The anti-inflammatory effect on contact dermatitis of the water solubilized 1'-Acetoxychavicol Acetate (ACA) by complexation with β-1,3-glucan isolated form Aureobasidium pullulans black yeast is reported. It is well-known that ACA possesses a function to inhibit the activation of NF-κB by which genes encoding proinflammatory cytokines, chemokines, and growth factors are regulated. However, because ACA is quite insoluble in water, its usefulness has been extremely limited. On the other hand, a triple-helical polysaccharide β-1,3-glucan can include hydrophobic compounds into intrastrand hydrophobic cavity and solubilize poorly water-soluble compounds. In this study, solubilization of ACA by complexation with highly branched β-1,3-glucan was achieved. The effect of anti-inflammatory response of water-soluble ACA complex with β-1,3-glucan was confirmed in vitro and in vivo.

  17. Fungal endophytes in germinated seeds of the common bean, Phaseolus vulgaris.

    PubMed

    Parsa, Soroush; García-Lemos, Adriana M; Castillo, Katherine; Ortiz, Viviana; López-Lavalle, Luis Augusto Becerra; Braun, Jerome; Vega, Fernando E

    2016-05-01

    We conducted a survey of fungal endophytes in 582 germinated seeds belonging to 11 Colombian cultivars of the common bean (Phaseolus vulgaris). The survey yielded 394 endophytic isolates belonging to 42 taxa, as identified by sequence analysis of the ribosomal DNA internal transcribed spacer (ITS) region. Aureobasidium pullulans was the dominant endophyte, isolated from 46.7 % of the samples. Also common were Fusarium oxysporum, Xylaria sp., and Cladosporium cladosporioides, but found in only 13.4 %, 11.7 %, and 7.6 % of seedlings, respectively. Endophytic colonization differed significantly among common bean cultivars and seedling parts, with the highest colonization occurring in the first true leaves of the seedlings. PMID:27109374

  18. Optimised method for the analysis of phenolic compounds from caper (Capparis spinosa L.) berries and monitoring of their changes during fermentation.

    PubMed

    Francesca, Nicola; Barbera, Marcella; Martorana, Alessandra; Saiano, Filippo; Gaglio, Raimondo; Aponte, Maria; Moschetti, Giancarlo; Settanni, Luca

    2016-04-01

    In this work, an ad hoc method to identify and quantify polyphenols from caper berries was developed on high-performance liquid chromatography/electrospray ionisation source/mass spectrometry (HPLC-ESI-MS). The method was applied during fermentation carried out with Lactobacillus pentosus OM13 (Trial S) and without starter (Trial C). A total of five polyphenols were identified. All samples contained high concentrations of rutin. Epicatechin was found in untreated fruits, on the contrary quercetin was detected during fermentation. Trial S was characterised by a more rapid acidification and lower levels of spoilage microorganisms than Trial C. L. pentosus dominated among the microbial community of both trials and the highest biodiversity, in terms of strains, was displayed by Trial C. Aureobasidium pullulans was the only yeast species found. The analytical method proposed allowed a high polyphenolic compound recovery from untreated and processed caper berries in short time. The starter culture reduced the bitter taste of the final product.

  19. Optimised method for the analysis of phenolic compounds from caper (Capparis spinosa L.) berries and monitoring of their changes during fermentation.

    PubMed

    Francesca, Nicola; Barbera, Marcella; Martorana, Alessandra; Saiano, Filippo; Gaglio, Raimondo; Aponte, Maria; Moschetti, Giancarlo; Settanni, Luca

    2016-04-01

    In this work, an ad hoc method to identify and quantify polyphenols from caper berries was developed on high-performance liquid chromatography/electrospray ionisation source/mass spectrometry (HPLC-ESI-MS). The method was applied during fermentation carried out with Lactobacillus pentosus OM13 (Trial S) and without starter (Trial C). A total of five polyphenols were identified. All samples contained high concentrations of rutin. Epicatechin was found in untreated fruits, on the contrary quercetin was detected during fermentation. Trial S was characterised by a more rapid acidification and lower levels of spoilage microorganisms than Trial C. L. pentosus dominated among the microbial community of both trials and the highest biodiversity, in terms of strains, was displayed by Trial C. Aureobasidium pullulans was the only yeast species found. The analytical method proposed allowed a high polyphenolic compound recovery from untreated and processed caper berries in short time. The starter culture reduced the bitter taste of the final product. PMID:26593604

  20. [Allergic alveolitis following inhalation of mould spores from pot plant earth (author's transl)].

    PubMed

    Velcovsky, H G; Graubner, M

    1981-01-23

    Acute exogenous allergic alveolitis with the typical symptoms of unproductive cough, dyspnoea on exertion, fever, loss of weight, headache and limb pains was observed in a 24-year-old bank employee. Leucocytosis was accompanied by raised ESR; pronounced hypoxaemia and marked restrictive ventilatory defects were found. X-ray changes consisted of severely increased fine reticular interstitial shadowing. The differential diagnosis of allergic alveolitis was confirmed by the demonstration of precipitating antibodies in serum against Pullularia pullulans, Trichoderma viride, Cephalosporium acremonium and Aspergillus fumigatus, moulds with an ubiquitous occurrence. Typical changes of alveolitis were found histopathologically and immunohistologically in the lung tissue after a mini-thoracotomy. The source of exposition leading to the disease were pot plant earth and containers in the patient's flat. Moulds could be isolated from these substrates.

  1. House Fly (Musca domestica L.) Attraction to Insect Honeydew.

    PubMed

    Hung, Kim Y; Michailides, Themis J; Millar, Jocelyn G; Wayadande, Astri; Gerry, Alec C

    2015-01-01

    House flies are of major concern as vectors of food-borne pathogens to food crops. House flies are common pests on cattle feedlots and dairies, where they develop in and feed on animal waste. By contacting animal waste, house flies can acquire human pathogenic bacteria such as Escherichia coli and Salmonella spp., in addition to other bacteria, viruses, or parasites that may infect humans and animals. The subsequent dispersal of house flies from animal facilities to nearby agricultural fields containing food crops may lead to pre-harvest food contamination with these pathogens. We hypothesized that odors from honeydew, the sugary excreta produced by sucking insects feeding on crops, or molds and fungi growing on honeydew, may attract house flies, thereby increasing the risk of food crop contamination. House fly attraction to honeydew-contaminated plant material was evaluated using a laboratory bioassay. House flies were attracted to the following plant-pest-honeydew combinations: citrus mealybug on squash fruit, pea aphid on faba bean plants, whitefly on navel orange and grapefruit leaves, and combined citrus mealybug and cottony cushion scale on mandarin orange leaves. House flies were not attracted to field-collected samples of lerp psyllids on eucalyptus plants or aphids on crepe myrtle leaves. Fungi associated with field-collected honeydews were isolated and identified for further study as possible emitters of volatiles attractive to house flies. Two fungal species, Aureobasidium pullulans and Cladosporium cladosporioides, were repeatedly isolated from field-collected honeydew samples. Both fungal species were grown in potato dextrose enrichment broth and house fly attraction to volatiles from these fungal cultures was evaluated. House flies were attracted to odors from A. pullulans cultures but not to those of C. cladosporioides. Identification of specific honeydew odors that are attractive to house flies could be valuable for the development of improved house

  2. Capillary Force Seeding of Hydrogels for Adipose-Derived Stem Cell Delivery in Wounds

    PubMed Central

    Garg, Ravi K.; Rennert, Robert C.; Duscher, Dominik; Sorkin, Michael; Kosaraju, Revanth; Auerbach, Lauren J.; Lennon, James; Chung, Michael T.; Paik, Kevin; Nimpf, Johannes; Rajadas, Jayakumar; Longaker, Michael T.

    2014-01-01

    Effective skin regeneration therapies require a successful interface between progenitor cells and biocompatible delivery systems. We previously demonstrated the efficiency of a biomimetic pullulan-collagen hydrogel scaffold for improving bone marrow-derived mesenchymal stem cell survival within ischemic skin wounds by creating a “stem cell niche” that enhances regenerative cytokine secretion. Adipose-derived mesenchymal stem cells (ASCs) represent an even more appealing source of stem cells because of their abundance and accessibility, and in this study we explored the utility of ASCs for hydrogel-based therapies. To optimize hydrogel cell seeding, a rapid, capillary force-based approach was developed and compared with previously established cell seeding methods. ASC viability and functionality following capillary hydrogel seeding were then analyzed in vitro and in vivo. In these experiments, ASCs were seeded more efficiently by capillary force than by traditional methods and remained viable and functional in this niche for up to 14 days. Additionally, hydrogel seeding of ASCs resulted in the enhanced expression of multiple stemness and angiogenesis-related genes, including Oct4, Vegf, Mcp-1, and Sdf-1. Moving in vivo, hydrogel delivery improved ASC survival, and application of both murine and human ASC-seeded hydrogels to splinted murine wounds resulted in accelerated wound closure and increased vascularity when compared with control wounds treated with unseeded hydrogels. In conclusion, capillary seeding of ASCs within a pullulan-collagen hydrogel bioscaffold provides a convenient and simple way to deliver therapeutic cells to wound environments. Moreover, ASC-seeded constructs display a significant potential to accelerate wound healing that can be easily translated to a clinical setting. PMID:25038246

  3. Culturable fungi of stored 'golden delicious' apple fruits: a one-season comparison study of organic and integrated production systems in Switzerland.

    PubMed

    Granado, José; Thürig, Barbara; Kieffer, Edith; Petrini, Liliane; Fliessbach, Andreas; Tamm, Lucius; Weibel, Franco P; Wyss, Gabriela S

    2008-11-01

    The effects of organic and integrated production systems on the culturable fungal microflora of stored apple fruits from five matched pairs of certified organic and integrated 'Golden Delicious' farms were studied at five representative production sites in Switzerland. Isolated fungi were identified morphologically. Colonization frequency (percentage of apples colonized), abundance (colony numbers), and diversity (taxon richness) were assessed for each orchard. The standard quality of the stored fruits was comparable for both organic and integrated apples and complied with national food hygiene standards. Yeasts (six taxa) and the yeast-like fungus Aureobasidium pullulans were the dominant epiphytes, filamentous fungi (21 taxa) the dominant endophytes. The most common fungi occurred at all sites and belonged to the "white" and "pink" yeasts, yeast-like A. pullulans, filamentous fungi Cladosporium spp., Alternaria spp., and sterile filamentous fungi. Canonical correspondence analysis of the total fungal community revealed a clear differentiation among production systems and sites. Compared to integrated apples, organic apples had significantly higher frequencies of filamentous fungi, abundance of total fungi, and taxon diversity. The effects of the production system on the fungal microflora are most likely due to the different plant protection strategies. The incidence of potential mycotoxin producers such as Penicillium and Alternaria species was not different between production systems. We suggest that higher fungal diversity may generally be associated with organic production and may increase the level of beneficial and antagonistically acting species known for their potential to suppress apple pathogens, which may be an advantage to organic apples, e.g., in respect to natural disease control. PMID:18473135

  4. Novel Symbiotic Protoplasts Formed by Endophytic Fungi Explain Their Hidden Existence, Lifestyle Switching, and Diversity within the Plant Kingdom

    PubMed Central

    Atsatt, Peter R.; Whiteside, Matthew D.

    2014-01-01

    Diverse fungi live all or part of their life cycle inside plants as asymptomatic endophytes. While endophytic fungi are increasingly recognized as significant components of plant fitness, it is unclear how they interact with plant cells; why they occur throughout the fungal kingdom; and why they are associated with most fungal lifestyles. Here we evaluate the diversity of endophytic fungi that are able to form novel protoplasts called mycosomes. We found that mycosomes cultured from plants and phylogenetically diverse endophytic fungi have common morphological characteristics, express similar developmental patterns, and can revert back to the free-living walled state. Observed with electron microscopy, mycosome ontogeny within Aureobasidium pullulans may involve two organelles: double membrane-bounded promycosome organelles (PMOs) that form mycosomes, and multivesicular bodies that may form plastid-infecting vesicles. Cultured mycosomes also contain a double membrane-bounded organelle, which may be homologous to the A. pullulans PMO. The mycosome PMO is often expressed as a vacuole-like organelle, which alternatively may contain a lipoid body or a starch grain. Mycosome reversion to walled cells occurs within the PMO, and by budding from lipid or starch-containing mycosomes. Mycosomes discovered in chicken egg yolk provided a plant-independent source for analysis: they formed typical protoplast stages, contained fungal ITS sequences and reverted to walled cells, suggesting mycosome symbiosis with animals as well as plants. Our results suggest that diverse endophytic fungi express a novel protoplast phase that can explain their hidden existence, lifestyle switching, and diversity within the plant kingdom. Importantly, our findings outline “what, where, when and how”, opening the way for cell and organelle-specific tests using in situ DNA hybridization and fluorescent labels. We discuss developmental, ecological and evolutionary contexts that provide a robust

  5. Enzyme activities in the Delaware Estuary affected by elevated suspended sediment load

    NASA Astrophysics Data System (ADS)

    Ziervogel, K.; Arnosti, C.

    2009-09-01

    Extracellular enzyme activities were compared among surface water, bottom water, and sediments of the Delaware Estuary using six fluorescently labeled, structurally distinct polysaccharides to determine the effects of suspended sediment transport on water column hydrolytic activities. Potential hydrolysis rates in surface waters were also measured for the nearby shelf. Samples were taken in December 2006, 6 months after a major flood event in the Delaware Basin that was followed by high freshwater run-off throughout the fall of 2006. All substrates were hydrolyzed in sediments and in the water column, including two (pullulan and fucoidan) that previously were not hydrolyzed in surface waters of the Delaware estuary. At the time of sampling, total particulate matter (TPM) in surface waters at the lower bay, bay mouth, and shelf ranged between 31 mg l -1 and 48 mg l -1 and were 2 to 20 times higher than previously reported. The presence of easily resuspended sediments at the lower bay and bay mouth indicated enhanced suspended sediment transport in the estuary prior to our sampling. Bottom water hydrolysis rates at the two sites affected by sediment resuspension were generally higher than those in surface waters from the same site. Most notably, fucoidan and pullulan hydrolysis rates in bay mouth bottom waters were 22.6 and 6.2 nM monomer h -1, respectively, and thus three and five times higher than surface water rates. Our data suggest that enhanced mixing processes between the sediment and the overlying water broadened the spectrum of water column hydrolases activity, improving the efficiency of enzymatic degradation of high molecular weight organic matter in the water with consequences for organic matter cycling in the Delaware estuary.

  6. House Fly (Musca domestica L.) Attraction to Insect Honeydew

    PubMed Central

    Hung, Kim Y.; Michailides, Themis J.; Millar, Jocelyn G.; Wayadande, Astri; Gerry, Alec C.

    2015-01-01

    House flies are of major concern as vectors of food-borne pathogens to food crops. House flies are common pests on cattle feedlots and dairies, where they develop in and feed on animal waste. By contacting animal waste, house flies can acquire human pathogenic bacteria such as Escherichia coli and Salmonella spp., in addition to other bacteria, viruses, or parasites that may infect humans and animals. The subsequent dispersal of house flies from animal facilities to nearby agricultural fields containing food crops may lead to pre-harvest food contamination with these pathogens. We hypothesized that odors from honeydew, the sugary excreta produced by sucking insects feeding on crops, or molds and fungi growing on honeydew, may attract house flies, thereby increasing the risk of food crop contamination. House fly attraction to honeydew-contaminated plant material was evaluated using a laboratory bioassay. House flies were attracted to the following plant-pest-honeydew combinations: citrus mealybug on squash fruit, pea aphid on faba bean plants, whitefly on navel orange and grapefruit leaves, and combined citrus mealybug and cottony cushion scale on mandarin orange leaves. House flies were not attracted to field-collected samples of lerp psyllids on eucalyptus plants or aphids on crepe myrtle leaves. Fungi associated with field-collected honeydews were isolated and identified for further study as possible emitters of volatiles attractive to house flies. Two fungal species, Aureobasidium pullulans and Cladosporium cladosporioides, were repeatedly isolated from field-collected honeydew samples. Both fungal species were grown in potato dextrose enrichment broth and house fly attraction to volatiles from these fungal cultures was evaluated. House flies were attracted to odors from A. pullulans cultures but not to those of C. cladosporioides. Identification of specific honeydew odors that are attractive to house flies could be valuable for the development of improved house

  7. Nanogel-crosslinked nanoparticles increase the inhibitory effects of W9 synthetic peptide on bone loss in a murine bone resorption model.

    PubMed

    Sato, Toshimi; Alles, Neil; Khan, Masud; Nagano, Kenichi; Takahashi, Mariko; Tamura, Yukihiko; Shimoda, Asako; Ohya, Keiichi; Akiyoshi, Kazunari; Aoki, Kazuhiro

    2015-01-01

    We investigated the biological activity of W9, a bone resorption inhibitor peptide, using NanoClik nanoparticles as an injectable carrier, where acryloyl group-modified cholesterol-bearing pullulan (CHPOA) nanogels were crosslinked by pentaerythritol tetra (mercaptoethyl) polyoxyethylene. Thirty 5-week-old male C57BL/6J mice were fed a low calcium diet and received once-daily subcutaneous injections of the carrier alone, W9 24 mg/kg/day alone, W9 24 mg/kg/day incorporated in cholesterol bearing pullulan (CHP) nanogels, or W9 (8 and 24 mg/kg/day) incorporated in NanoClik nanoparticles for 4 days (n=5). Mice that received a normal calcium diet with NanoClik nanoparticle injections without W9 were used as a control group. Radiological analyses showed that administration of W9 24 mg/kg/day significantly prevented low calcium-induced reduction of bone mineral density in the long bones and lumbar vertebrae, but only when the NanoClik nanoparticles were used as a carrier. Histomorphometric analyses of the proximal tibiae revealed that W9 24 mg/kg/day incorporated in NanoClik nanoparticles prevented the increase in bone resorption indices induced by a low calcium diet, which was confirmed by measurement of serum bone resorption markers. These data suggest that NanoClik nanoparticles could be a useful carrier for peptide therapeutics, and also demonstrate that daily subcutaneous injections of the W9 peptide with the nanoparticles were able to inhibit bone loss in vivo. An osteoclastogenesis inhibition assay performed in vitro confirmed a slower release profile of W9 from NanoClik nanoparticles compared with conventional CHP nanogels. PMID:25999711

  8. Novel symbiotic protoplasts formed by endophytic fungi explain their hidden existence, lifestyle switching, and diversity within the plant kingdom.

    PubMed

    Atsatt, Peter R; Whiteside, Matthew D

    2014-01-01

    Diverse fungi live all or part of their life cycle inside plants as asymptomatic endophytes. While endophytic fungi are increasingly recognized as significant components of plant fitness, it is unclear how they interact with plant cells; why they occur throughout the fungal kingdom; and why they are associated with most fungal lifestyles. Here we evaluate the diversity of endophytic fungi that are able to form novel protoplasts called mycosomes. We found that mycosomes cultured from plants and phylogenetically diverse endophytic fungi have common morphological characteristics, express similar developmental patterns, and can revert back to the free-living walled state. Observed with electron microscopy, mycosome ontogeny within Aureobasidium pullulans may involve two organelles: double membrane-bounded promycosome organelles (PMOs) that form mycosomes, and multivesicular bodies that may form plastid-infecting vesicles. Cultured mycosomes also contain a double membrane-bounded organelle, which may be homologous to the A. pullulans PMO. The mycosome PMO is often expressed as a vacuole-like organelle, which alternatively may contain a lipoid body or a starch grain. Mycosome reversion to walled cells occurs within the PMO, and by budding from lipid or starch-containing mycosomes. Mycosomes discovered in chicken egg yolk provided a plant-independent source for analysis: they formed typical protoplast stages, contained fungal ITS sequences and reverted to walled cells, suggesting mycosome symbiosis with animals as well as plants. Our results suggest that diverse endophytic fungi express a novel protoplast phase that can explain their hidden existence, lifestyle switching, and diversity within the plant kingdom. Importantly, our findings outline "what, where, when and how", opening the way for cell and organelle-specific tests using in situ DNA hybridization and fluorescent labels. We discuss developmental, ecological and evolutionary contexts that provide a robust

  9. Selection and use of pectinolytic yeasts for improving clarification and phenolic extraction in winemaking.

    PubMed

    Belda, Ignacio; Conchillo, Lorena B; Ruiz, Javier; Navascués, Eva; Marquina, Domingo; Santos, Antonio

    2016-04-16

    Pectinase enzymes have shown a considerable influence in both, sensitive and technological properties of wines. They can help to improve clarification process, releasing more color and flavor compounds entrapped in grape skin, facilitating the liberation of phenolic compounds. This work aims to find yeasts that, because of their native pectinases, can be applied on combined fermentations with Saccharomyces cerevisiae obtaining significant benefits over single-inoculated traditional fermentations. 462 yeast strains isolated from wineries were identified and tested for several enzymatic activities of recognized interest for enology industry. Considering the 7 identified species, only Aureobasidium pullulans, Metschnikowia pulcherrima and Metschnikowia fructicola showed polygalacturonase activity. Because of its interest in winemaking, due to its reported incidence in wine flavor, the impact of M. pulcherrima as a source of pectinolytic enzymes was analyzed by measuring its influence in filterability, turbidity and the increase on color, anthocyanin and polyphenol content of wines fermented in combination with S. cerevisiae. Among the strains screened, M. pulcherrima NS-EM-34 was selected, due to its polygalacturonase activity, for further characterization in both, laboratory and semi-industrial scale assays. The kinetics concerning several metabolites of enological concern were followed during the entire fermentation process at microvinification scale. Improved results were obtained in the expected parameters when M. pulcherrima NS-EM-34 was used, in comparison to wines fermented with S. cerevisiae alone and combined with other pectinolytic and non-pectinolytic yeasts (A. pullulans and Lachancea thermotolerans, respectively), even working better than commercial enzymes preparations in most parameters. Additionally, M. pulcherrima NS-EM-34 was used at a semi-industrial scale combined with three different S. cerevisiae strains, confirming its potential application for

  10. Nanogel-crosslinked nanoparticles increase the inhibitory effects of W9 synthetic peptide on bone loss in a murine bone resorption model

    PubMed Central

    Sato, Toshimi; Alles, Neil; Khan, Masud; Nagano, Kenichi; Takahashi, Mariko; Tamura, Yukihiko; Shimoda, Asako; Ohya, Keiichi; Akiyoshi, Kazunari; Aoki, Kazuhiro

    2015-01-01

    We investigated the biological activity of W9, a bone resorption inhibitor peptide, using NanoClik nanoparticles as an injectable carrier, where acryloyl group-modified cholesterol-bearing pullulan (CHPOA) nanogels were crosslinked by pentaerythritol tetra (mercaptoethyl) polyoxyethylene. Thirty 5-week-old male C57BL/6J mice were fed a low calcium diet and received once-daily subcutaneous injections of the carrier alone, W9 24 mg/kg/day alone, W9 24 mg/kg/day incorporated in cholesterol bearing pullulan (CHP) nanogels, or W9 (8 and 24 mg/kg/day) incorporated in NanoClik nanoparticles for 4 days (n=5). Mice that received a normal calcium diet with NanoClik nanoparticle injections without W9 were used as a control group. Radiological analyses showed that administration of W9 24 mg/kg/day significantly prevented low calcium-induced reduction of bone mineral density in the long bones and lumbar vertebrae, but only when the NanoClik nanoparticles were used as a carrier. Histomorphometric analyses of the proximal tibiae revealed that W9 24 mg/kg/day incorporated in NanoClik nanoparticles prevented the increase in bone resorption indices induced by a low calcium diet, which was confirmed by measurement of serum bone resorption markers. These data suggest that NanoClik nanoparticles could be a useful carrier for peptide therapeutics, and also demonstrate that daily subcutaneous injections of the W9 peptide with the nanoparticles were able to inhibit bone loss in vivo. An osteoclastogenesis inhibition assay performed in vitro confirmed a slower release profile of W9 from NanoClik nanoparticles compared with conventional CHP nanogels. PMID:25999711

  11. House Fly (Musca domestica L.) Attraction to Insect Honeydew.

    PubMed

    Hung, Kim Y; Michailides, Themis J; Millar, Jocelyn G; Wayadande, Astri; Gerry, Alec C

    2015-01-01

    House flies are of major concern as vectors of food-borne pathogens to food crops. House flies are common pests on cattle feedlots and dairies, where they develop in and feed on animal waste. By contacting animal waste, house flies can acquire human pathogenic bacteria such as Escherichia coli and Salmonella spp., in addition to other bacteria, viruses, or parasites that may infect humans and animals. The subsequent dispersal of house flies from animal facilities to nearby agricultural fields containing food crops may lead to pre-harvest food contamination with these pathogens. We hypothesized that odors from honeydew, the sugary excreta produced by sucking insects feeding on crops, or molds and fungi growing on honeydew, may attract house flies, thereby increasing the risk of food crop contamination. House fly attraction to honeydew-contaminated plant material was evaluated using a laboratory bioassay. House flies were attracted to the following plant-pest-honeydew combinations: citrus mealybug on squash fruit, pea aphid on faba bean plants, whitefly on navel orange and grapefruit leaves, and combined citrus mealybug and cottony cushion scale on mandarin orange leaves. House flies were not attracted to field-collected samples of lerp psyllids on eucalyptus plants or aphids on crepe myrtle leaves. Fungi associated with field-collected honeydews were isolated and identified for further study as possible emitters of volatiles attractive to house flies. Two fungal species, Aureobasidium pullulans and Cladosporium cladosporioides, were repeatedly isolated from field-collected honeydew samples. Both fungal species were grown in potato dextrose enrichment broth and house fly attraction to volatiles from these fungal cultures was evaluated. House flies were attracted to odors from A. pullulans cultures but not to those of C. cladosporioides. Identification of specific honeydew odors that are attractive to house flies could be valuable for the development of improved house

  12. Nanogel-crosslinked nanoparticles increase the inhibitory effects of W9 synthetic peptide on bone loss in a murine bone resorption model.

    PubMed

    Sato, Toshimi; Alles, Neil; Khan, Masud; Nagano, Kenichi; Takahashi, Mariko; Tamura, Yukihiko; Shimoda, Asako; Ohya, Keiichi; Akiyoshi, Kazunari; Aoki, Kazuhiro

    2015-01-01

    We investigated the biological activity of W9, a bone resorption inhibitor peptide, using NanoClik nanoparticles as an injectable carrier, where acryloyl group-modified cholesterol-bearing pullulan (CHPOA) nanogels were crosslinked by pentaerythritol tetra (mercaptoethyl) polyoxyethylene. Thirty 5-week-old male C57BL/6J mice were fed a low calcium diet and received once-daily subcutaneous injections of the carrier alone, W9 24 mg/kg/day alone, W9 24 mg/kg/day incorporated in cholesterol bearing pullulan (CHP) nanogels, or W9 (8 and 24 mg/kg/day) incorporated in NanoClik nanoparticles for 4 days (n=5). Mice that received a normal calcium diet with NanoClik nanoparticle injections without W9 were used as a control group. Radiological analyses showed that administration of W9 24 mg/kg/day significantly prevented low calcium-induced reduction of bone mineral density in the long bones and lumbar vertebrae, but only when the NanoClik nanoparticles were used as a carrier. Histomorphometric analyses of the proximal tibiae revealed that W9 24 mg/kg/day incorporated in NanoClik nanoparticles prevented the increase in bone resorption indices induced by a low calcium diet, which was confirmed by measurement of serum bone resorption markers. These data suggest that NanoClik nanoparticles could be a useful carrier for peptide therapeutics, and also demonstrate that daily subcutaneous injections of the W9 peptide with the nanoparticles were able to inhibit bone loss in vivo. An osteoclastogenesis inhibition assay performed in vitro confirmed a slower release profile of W9 from NanoClik nanoparticles compared with conventional CHP nanogels.

  13. 75 FR 21567 - Federal Motor Vehicle Safety Standard No. 108; Lamp, Reflective Devices and Associated Equipment...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-04-26

    ... amended standard, this requirement is found in paragraph S10.17.1.1.2 and paragraph S10.17.1.1.3. See 72 FR 68234 (Dec. 4, 2007). In a December 2007 final rule, NHTSA rewrote and reorganized FMVSS No. 108.... Id. The effective date of those amendments has been delayed to December 1, 2012. 74 FR 58214 (Nov....

  14. 78 FR 60928 - Request To Amend a License To Export High-Enriched Uranium

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-10-02

    ... COMMISSION Request To Amend a License To Export High-Enriched Uranium Pursuant to 10 CFR 110.70 (b) ``Public... Nuclear Security Uranium uranium (17.1 targets in France Administration, September 9, (93.35%). kilograms... 10.1 kg uranium to a new cumulative total of 17.1 kg of U-235 contained in 18.4 kg uranium; and...

  15. Application of exopolysaccharides to improve the performance of ceramic bodies in the unidirectional dry pressing process

    NASA Astrophysics Data System (ADS)

    Caneira, Inês; Machado-Moreira, Bernardino; Dionísio, Amélia; Godinho, Vasco; Neves, Orquídia; Dias, Diamantino; Saiz-Jimenez, Cesareo; Miller, Ana Z.

    2015-04-01

    Ceramic industry represents an important sector of economic activity in the European countries and involves complex and numerous manufacturing processes. The unidirectional dry pressing process includes milling and stirring of raw materials (mainly clay and talc minerals) in aqueous suspensions, followed by spray drying to remove excess water obtaining spray-dried powders further subjected to dry pressing process (conformation). However, spray-dried ceramic powders exhibit an important variability in their performance when subjected to the dry pressing process, particularly in the adhesion to the mold and mechanical strength, affecting the quality of the final conformed ceramic products. Therefore, several synthetic additives (deflocculants, antifoams, binders, lubricants and plasticizers) are introduced in the ceramic slips to achieve uniform and homogeneous pastes, conditioning their rheological properties. However, an important variability associated with the performance of the conformed products is still reported. Exopolysaccharides or Extracellular Polymeric Substances (EPS) are polymers excreted by living organisms, such as bacteria, fungi and algae, which may confer unique and potentially interesting properties with potential industrial uses, such as viscosity control, gelation, and flocculation. Polysaccharides, such as pullulan, gellan, carrageenan and xanthan have found a wide range of applications in food, pharmaceutical, petroleum, and in other industries. The aim of this study was the assessment of exopolysaccharides as natural additives to optimize the performance of spray-dried ceramic powders during the unidirectional dry pressing process, replacing the synthetic additives used in the ceramic production process. Six exopolysaccharides, namely pullulan, gellan, xanthan gum, κappa- and iota-carrageenan, and guar gum were tested in steatite-based spray-dried ceramic powders at different concentrations. Subsequently, these ceramic powders were

  16. Measles

    MedlinePlus

    ... 2000. During 2000-2014, measles vaccination prevented an estimated 17.1 million deaths making measles vaccine one ... In 1980, before widespread vaccination, measles caused an estimated 2.6 million deaths each year. The disease ...

  17. (Genetic engineering with a gene encoding a soybean storage protein). Progress report

    SciTech Connect

    Beachy, R.N.

    1985-01-01

    Progress is reported on research directed toward introducing a gene (Gmg 17.1) encoding the ..cap alpha..'-subunit of ..beta..-conglycinin, a soybean seed protein, into petunia plants using gene transfer mechanisms. (ACR)

  18. 44 CFR 206.119 - Financial assistance to address other needs.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ...) established under the National Flood Insurance Program (NFIP) regulations at 44 CFR 61.17. (1) The premium for... generally limited to the following (i) Funeral services; (ii) Burial or cremation; and (iii) Other...

  19. 44 CFR 206.119 - Financial assistance to address other needs.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ...) established under the National Flood Insurance Program (NFIP) regulations at 44 CFR 61.17. (1) The premium for... generally limited to the following (i) Funeral services; (ii) Burial or cremation; and (iii) Other...

  20. 37 CFR 1.451 - The priority claim and priority document in an international application.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... the International Bureau (PCT Article 8 and PCT Rule 17). The fee for preparing a certified copy is... the United States Receiving Office within the time limit specified in PCT Rule 17.1(a). (d)...

  1. 40 CFR 86.528-78 - Transmissions.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... when engine stalling is imminent. (g) If downshifting during deceleration is not permitted in the... displacements less than 280 cc (17.1 cu. in.), when engine roughness is evident, or when engine stalling...

  2. 40 CFR 86.419-78 - Engine displacement, motorcycle classes.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... rotors. cc=(max. chamber volume − min. chamber volume) × 3 × no. of rotors (b) Motorcycles will be divided into classes based on engine displacement. (1) Class I—50 to 169 cc (3.1 to 10.4 cu. in.). (2) Class II—170 to 279 cc (10.4 to 17.1 cu. in.). (3) Class III—280 cc and over (17.1 cu. in. and over)....

  3. The Effect of Optional Summer Transition to High School Program Participation and Required School Year Long Placement on a Core Content Team on Ninth-Grade Students At-Risk Measured Achievement, Engagement, and Behavior Outcomes

    ERIC Educational Resources Information Center

    Pokorski, Frances K.

    2011-01-01

    No significant difference in beginning ninth-grade pretest compared to ending ninth-grade posttest comparisons of American History t(17) = 0.34, p = 0.37 (one-tailed), d = 0.09, English t(17) = 1.40, p = 0.09 (one-tailed), d = 0.34, and Biology (t(17) = -1.58, p = 0.07 (one-tailed), d = -0.22, course grade scores were observed for students…

  4. 40 CFR 180.585 - Pyraflufen-ethyl; tolerances for residues.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... Goat, meat byproducts 0.03 Grape 0.01 Grass, forage, group 17 1.0 Grass, hay, group 17 1.4 Horse, fat 0.03 Horse, meat 0.03 Horse, meat byproducts 0.03 Milk 0.03 Nut, tree, group 14 0.01 Olive 0.01 Peanut... million Almond, hulls 0.02 Cattle, fat 0.03 Cattle, meat 0.03 Cattle, meat byproducts 0.03 Corn,...

  5. 40 CFR 180.585 - Pyraflufen-ethyl; tolerances for residues.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... Goat, meat byproducts 0.03 Grape 0.01 Grass, forage, group 17 1.0 Grass, hay, group 17 1.4 Horse, fat 0.03 Horse, meat 0.03 Horse, meat byproducts 0.03 Milk 0.03 Nut, tree, group 14 0.01 Olive 0.01 Peanut... million Almond, hulls 0.02 Cattle, fat 0.03 Cattle, meat 0.03 Cattle, meat byproducts 0.03 Corn,...

  6. 50 CFR 665.269 - Quotas.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... Beds Au'au Channel Black: 5,000 2 Makapu'u Pink: 2,000 2 Gold: 0 (zero) Bamboo: 500 2 Conditional Beds 180 Fathom Bank Pink: 222 1 Gold: 67 1 Bamboo: 56 1 Brooks Bank Pink: 444 1 Gold: 133 1 Bamboo: 111 1 Kaena Point Pink: 67 1 Gold: 20 1 Bamboo: 17 1 Keahole Point Pink: 67 1 Gold: 20 1 Bamboo: 17 1...

  7. 50 CFR 665.269 - Quotas.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... Beds Au'au Channel Black: 5,000 2 Makapu'u Pink: 2,000 2 Gold: 0 (zero) Bamboo: 500 2 Conditional Beds 180 Fathom Bank Pink: 222 1 Gold: 67 1 Bamboo: 56 1 Brooks Bank Pink: 444 1 Gold: 133 1 Bamboo: 111 1 Kaena Point Pink: 67 1 Gold: 20 1 Bamboo: 17 1 Keahole Point Pink: 67 1 Gold: 20 1 Bamboo: 17 1...

  8. 50 CFR 665.269 - Quotas.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... Beds Au'au Channel Black: 5,000 2 Makapu'u Pink: 2,000 2 Gold: 0 (zero) Bamboo: 500 2 Conditional Beds 180 Fathom Bank Pink: 222 1 Gold: 67 1 Bamboo: 56 1 Brooks Bank Pink: 444 1 Gold: 133 1 Bamboo: 111 1 Kaena Point Pink: 67 1 Gold: 20 1 Bamboo: 17 1 Keahole Point Pink: 67 1 Gold: 20 1 Bamboo: 17 1...

  9. 50 CFR 665.269 - Quotas.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... Beds Au'au Channel Black: 5,000 2 Makapu'u Pink: 2,000 2 Gold: 0 (zero) Bamboo: 500 2 Conditional Beds 180 Fathom Bank Pink: 222 1 Gold: 67 1 Bamboo: 56 1 Brooks Bank Pink: 444 1 Gold: 133 1 Bamboo: 111 1 Kaena Point Pink: 67 1 Gold: 20 1 Bamboo: 17 1 Keahole Point Pink: 67 1 Gold: 20 1 Bamboo: 17 1...

  10. 50 CFR 665.269 - Quotas.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... Beds Au'au Channel Black: 5,000 2 Makapu'u Pink: 2,000 2 Gold: 0 (zero) Bamboo: 500 2 Conditional Beds 180 Fathom Bank Pink: 222 1 Gold: 67 1 Bamboo: 56 1 Brooks Bank Pink: 444 1 Gold: 133 1 Bamboo: 111 1 Kaena Point Pink: 67 1 Gold: 20 1 Bamboo: 17 1 Keahole Point Pink: 67 1 Gold: 20 1 Bamboo: 17 1...

  11. Molecular Phylogenetics of the Genus Trichosporon Inferred from Mitochondrial Cytochrome b Gene Sequences

    PubMed Central

    Biswas, Swarajit Kumar; Wang, Li; Yokoyama, Koji; Nishimura, Kazuko

    2005-01-01

    Mitochondrial cytochrome b (cyt b) genes of 42 strains representing 23 species of the genus Trichosporon were partially sequenced to determine their molecular phylogenetic relationships. Almost half of the 22 strains investigated (from 11 different species) contained introns in their sequences. Analysis of a 396-bp coding sequence from each strain of Trichosporon under investigation showed a total of 141 (35.6%) variable nucleotide sites. A phylogenetic tree based on the cyt b gene sequences revealed that all species of Trichosporon except Trichosporon domesticum and Trichosporon montevideense had species-specific cyt b genes. Trichosporon sp. strain CBS 5581 was identified as Trichosporon pullulans, and one clinical isolate, IFM 48794, was identified as Trichosporon faecale. Analysis of 132-bp deduced amino acid sequences showed a total of 34 (25.75%) variable amino acid sites. T. domesticum and T. montevideense, Trichosporon asahii and Trichosporon asteroides, and Trichosporon gracile and Trichosporon guehoae had identical amino acid sequences. A phylogenetic tree constructed with the ascomycetes Saccharomyces douglasii and Candida glabrata taken as outgroup species and including representative species from closely related genera species of Trichosporon clustered with other basidiomycetous yeasts that contain xylose in their cell wall compositions. These results indicate the effectiveness of mitochondrial cyt b gene sequences for both species identification and the phylogenetic analysis of Trichosporon species. PMID:16207980

  12. The diversity and antifungal susceptibility of the yeasts isolated from coconut water and reconstituted fruit juices in Brazil.

    PubMed

    Maciel, Natália O P; Piló, Fernanda B; Freitas, Larissa F D; Gomes, Fátima C O; Johann, Susana; Nardi, Regina M D; Lachance, Marc-André; Rosa, Carlos A

    2013-01-01

    The aims of this study were to characterise the yeasts present in the reconstituted fruit juices and coconut water extracted with "coconut machines", both collected from commercial outlets in a Brazilian city, and to investigate the antifungal resistance of isolates from these beverages that were able to grow at 37°C. The yeast population counts in the coconut water samples ranged from 1.7 to >6.5logcfu/ml, and in the reconstituted fruit juices, the counts ranged from 1.5 to >5.5logcfu/ml. Aureobasidium pullulans, Candida boidinii, Candidaintermedia, Candidaoleophila, Candidaparapsilosis, Candidasantamariae, Candidatropicalis, Clavispora lusitaniae, Kloeckera apis, Lachancea fermentati, Pichia fermentans and Rhodotorula mucilaginosa were the most frequent species isolated from these beverages. At least 18 yeast species isolated from these beverages have been reported as opportunistic pathogens. Eight yeast isolates were resistant to fluconazole, seven were resistant to itraconazole, and 26 to amphotericin B. Some yeast species were resistant to more than one of the antifungal drugs tested. Two isolates of C. tropicalis from the reconstituted fruit juices exhibited resistance to all three drugs. The presence of yeast strains that are resistant to commonly used antifungal drugs suggests a potential risk, at least to immunocompromised individuals who consume these beverages.

  13. Characterization of recombinant amylopullulanase (gt-apu) and truncated amylopullulanase (gt-apuT) of the extreme thermophile Geobacillus thermoleovorans NP33 and their action in starch saccharification.

    PubMed

    Nisha, M; Satyanarayana, T

    2013-07-01

    A gene encoding amylopullulanase (gt-apu) of the extremely thermophilic Geobacillus thermoleovorans NP33 was cloned and expressed in Escherichia coli. The gene has an open reading frame of 4,965 bp that encodes a protein of 1,655 amino acids with molecular mass of 182 kDa. The six conserved regions, characteristic of GH13 family, have been detected in gt-apu. The recombinant enzyme has only one active site for α-amylase and pullulanase activities based on the enzyme kinetic analyses in a system that contains starch as well as pullulan as competing substrates and response to inhibitors. The end-product analysis confirmed that this is an endoacting enzyme. The specific enzyme activities for α-amylase and pullulanase of the truncated amylopullulanase (gt-apuT) are higher than gt-apu. Both enzymes exhibited similar temperature (60 °C) and pH (7.0) optima, although gt-apuT possessed a higher thermostability than gt-apu. The overall catalytic efficiency (K(cat)/K(m)) of gt-apuT is greater than that of gt-apu, with almost similar substrate specificities. The C-terminal region of gt-apu appeared to be non-essential, and furthermore, it negatively affects the substrate binding and stability of the enzyme. PMID:23132347

  14. Membrane-bound amylopullulanase is essential for starch metabolism of Sulfolobus acidocaldarius DSM639.

    PubMed

    Choi, Kyoung-Hwa; Cha, Jaeho

    2015-09-01

    Sulfolobus acidocaldarius DSM639 produced an acid-resistant membrane-bound amylopullulanase (Apu) during growth on starch as a sole carbon and energy source. The physiological role of Apu in starch metabolism was investigated by the growth and starch degradation pattern of apu disruption mutant as well as biochemical properties of recombinant Apu. The Δapu mutant lost the ability to grow in minimal medium in the presence of starch, and the amylolytic activity observed in the membrane fraction of the wild-type strain was not detected in the Δapu mutant when the cells were grown in YT medium. The purified membrane-bound Apu initially hydrolyzed starch, amylopectin, and pullulan into various sizes of maltooligosaccharides, and then produced glucose, maltose, and maltotriose in the end, indicating Apu is a typical endo-acting glycoside hydrolase family 57 (GH57) amylopullulanase. The maltose and maltotriose observed in the culture medium during the exponential and stationary phase growth indicates that Apu is the essential enzyme to initially hydrolyze the starch into small maltooligosaccharides to be transported into the cell. PMID:26104674

  15. Characterization of an amylase produced by a Trichoderma harzianum isolate with antagonistic activity against Crinipellis perniciosa, the causal agent of witches' broom of cocoa.

    PubMed

    de Azevedo, A M; De Marco, J L; Felix, C R

    2000-07-15

    An isolate of Trichoderma harzianum showing antagonistic activity against Crinipellis perniciosa, the causal agent of the witches' broom disease of cocoa, produces substantial amounts of hydrolytic enzymes. An amylase purified from isolate 1051 had a molecular mass of about 68.7 kDa. Maximal activity against soluble starch was determined at pH 4.0 and 60 degrees C. The K(m) and V(max) values were 3.5 mg ml(-1) and 1.67 mg min(-1) of reducing sugar. The end products were mostly malto-oligosaccharides. The enzyme also hydrolyzed glycogen, amylopectin, maltotriose, and maltotetraose, but not pullulan or cellobiose. Maltose was only barely hydrolyzed. The purified amylase exerted a discrete hydrolytic effect on the C. perniciosa cell wall in vitro as observed by scanning electron microscopic analysis. While Fe(3+), Al(3+), Zn(2+), and Cu(2+) were effective in inhibiting the purified amylase, Mn(2+) considerably enhanced the activity. Ca(2+), Mg(2+), and Co(2+) showed no substantial effect on enzyme activity.

  16. Nanogel-based immunologically stealth vaccine targets macrophages in the medulla of lymph node and induces potent antitumor immunity.

    PubMed

    Muraoka, Daisuke; Harada, Naozumi; Hayashi, Tae; Tahara, Yoshiro; Momose, Fumiyasu; Sawada, Shin-ichi; Mukai, Sada-atsu; Akiyoshi, Kazunari; Shiku, Hiroshi

    2014-09-23

    Because existing therapeutic cancer vaccines provide only a limited clinical benefit, a different vaccination strategy is necessary to improve vaccine efficacy. We developed a nanoparticulate cancer vaccine by encapsulating a synthetic long peptide antigen within an immunologically inert nanoparticulate hydrogel (nanogel) of cholesteryl pullulan (CHP). After subcutaneous injection to mice, the nanogel-based vaccine was efficiently transported to the draining lymph node, and was preferentially engulfed by medullary macrophages but was not sensed by other macrophages and dendritic cells (so-called "immunologically stealth mode"). Although the function of medullary macrophages in T cell immunity has been unexplored so far, these macrophages effectively cross-primed the vaccine-specific CD8(+) T cells in the presence of a Toll-like receptor (TLR) agonist as an adjuvant. The nanogel-based vaccine significantly inhibited in vivo tumor growth in the prophylactic and therapeutic settings, compared to another vaccine formulation using a conventional delivery system, incomplete Freund's adjuvant. We also revealed that lymph node macrophages were highly responsive to TLR stimulation, which may underlie the potency of the macrophage-oriented, nanogel-based vaccine. These results indicate that targeting medullary macrophages using the immunologically stealth nanoparticulate delivery system is an effective vaccine strategy.

  17. Purification and characterization of camel (Camelus dromedarius) milk amylase.

    PubMed

    El-Fakharany, Esmail M; Serour, Ehab A; Abdelrahman, Aref M; Haroun, Bakry M; Redwan, El-Rashdy M

    2009-01-01

    Skimmed camel milk contains 59,900 U/L amylase, which is 39,363 times less than serum and plasma amylase. Camel milk beta-amylase was purified as a 61 KDa band using DEAE-Sepharose and Sephadex G-100 and yielded 561 U/mg. The optimum working pH, Km and temperature were 7.0, 13.6 mg/Lstarch, 30-40 degrees C, respectively. The enzyme has been shown higher affinity toward amylose and soluble starch than glycogen, amylopectin, dextrin, or pullulan. Magnesium chloride, CaCl(2) and NaCl activated the amylase, while EDTA and EGTA decreased its activity. While its activity was increased in the presence of Triton X-100 and Triton X-114. Phenylmethanesulfonyl fluoride did not show any effect on enzyme activity. However, the enzyme activity was inhibited by urea, SDS, DTNB, iodoacetamide, N-ethylmalimide, aprotinin, and trypsin inhibitor. It worked on starch to yield a maltose. Scanning electron microscope images demonstrated a nano-degrading ability on starch granules from various sources (potato, corn, cassava, and rice). PMID:19291574

  18. Polymeric photosensitizer-embedded self-expanding metal stent for repeatable endoscopic photodynamic therapy of cholangiocarcinoma.

    PubMed

    Bae, Byoung-chan; Yang, Su-Geun; Jeong, Seok; Lee, Don Haeng; Na, Kun; Kim, Joon Mee; Costamagna, Guido; Kozarek, Richard A; Isayama, Hiroyuki; Deviere, Jacques; Seo, Dong Wan; Nageshwar Reddy, D

    2014-10-01

    Photodynamic therapy (PDT) is a new therapeutic approach for the palliative treatment of malignant bile duct obstruction. In this study, we designed photosensitizer-embedded self-expanding nonvascular metal stent (PDT-stent) which allows repeatable photodynamic treatment of cholangiocarcinoma without systemic injection of photosensitizer. Polymeric photosensitizer (pullulan acetate-conjugated pheophorbide A; PPA) was incorporated in self-expanding nonvascular metal stent. Residence of PPA in the stent was estimated in buffer solution and subcutaneous implantation on mouse. Photodynamic activity of PDT-stent was evaluated through laserexposure on stent-layered tumor cell lines, HCT-116 tumor-xenograft mouse models and endoscopic intervention of PDT-stent on bile duct of mini pigs. Photo-fluorescence imaging of the PDT-stent demonstrated homogeneous embedding of polymeric Pheo-A (PPA) on stent membrane. PDT-stent sustained its photodynamic activities at least for 2 month. And which implies repeatable endoscopic PDT is possible after stent emplacement. The PDT-stent after light exposure successfully generated cytotoxic singlet oxygen in the surrounding tissues, inducing apoptotic degradation of tumor cells and regression of xenograft tumors on mouse models. Endoscopic biliary in-stent photodynamic treatments on minipigs also suggested the potential efficacy of PDT-stent on cholangiocarcinoma. In vivo and in vitro studies revealed our PDT-stent, allows repeatable endoscopic biliary PDT, has the potential for the combination therapy (stent plus PDT) of cholangiocarcinoma.

  19. Characterization of a pH and detergent-tolerant, cold-adapted type I pullulanase from Exiguobacterium sp. SH3.

    PubMed

    Rajaei, Sarah; Noghabi, Kambiz Akbari; Sadeghizadeh, Majid; Zahiri, Hossein Shahbani

    2015-11-01

    A pullulanase-encoding gene from psychrotrophic Exiguobacterium sp. SH3 was cloned and expressed in both E. coli and Bacillus subtilis. The functional recombinant enzyme (Pul-SH3) was purified as a His-tagged protein. Pul-SH3 was characterized to be a cold-adapted type I pullulanase with maximum activity at 45 °C. Using fluorescence spectroscopy, the melting temperature of Pul-SH3 was determined to be about 52 °C. The enzyme was able to hydrolyze pullulan, soluble starch, potato starch, and rice flour. It was exceptionally tolerant in the pH range of 4-11, exhibiting maximum activity at pH 8.5 and more than 60% of the activity in the pH range of 5-11. Being a detergent-tolerant pullulanase, Pul-SH3 retained 99, 89, and 54% of its activity at 10% concentration of Triton-X100, Tween 20, and SDS, respectively. The enzyme also exhibited an activity of 80.4 and 93.7% in the presence of two commercial detergents, Rika (7.5% v/v) and Fadisheh (2.5% w/v), respectively. The enzyme was even able to remain active by 54.5 and 85% after 10-day holding with the commercial detergents. Thermal stability of the enzyme could w on silica. Pul-SH3 with several industrially important characteristics seems desirable for cold hydrolysis of starch.

  20. Different drying technologies and alternation of mycobiots in the raw material of Hyssopus officinalis L.

    PubMed

    Raila, Algirdas; Lugauskas, Albinas; Kemzūraite, Aurelija; Zvicevicius, Egidijus; Ragazinskiene, Ona; Railiene, Marija

    2009-01-01

    Contamination of medicinal plant mass with mycobiots is one of the negative factors deteriorating the quality of raw material. In order to evaluate the impact of the yield processing technologies upon the changes of mycobiots in raw material, the mycobiotic conditions of herb hyssop (Hyssopus officinalis L.) raw material were evaluated under various regimes of active ventilation and optimization of the drying parameters. The impact of ventilation intensity and temperature of drying agent upon the changes and abundance of mycobiota species in medicinal raw material was determined. Irrespective of the temperature of the airflow, the strongest suppressive effect upon the mycobiotic contamination in Hyssopi herba was produced by the 5,000 m3 x (t x h)(-1) airflow. Analysis of the isolated fungi revealed the prevalence of Penicillium, Aspergillus, Alternaria, Cladosporium, Mucor, Rhizopus species in the raw material. In separate samples Botrytis cinerea, Sclerotinia sclerotiorum, Aureobasidium pullulans, Chrysosporium merdarium, Cladorrhinum foecundissimum, Ulocladium consortiale, Trichoderma hamatum, T. harzianum, Gilmaniella humicola, Talaromyces flavus, Rhizomucor pusillus, Hansfordia ovalispora, Verticicladium trifi dum, Trichosporiella cerebriformis micromycetes were also rather abundant. Detection of the above-mentioned micromycetes in herb hyssop samples differed, and partially depended upon the medium used for their isolation.

  1. Electrospun novel super-absorbent based on polysaccharide-polyvinyl alcohol-montmorillonite clay nanocomposites.

    PubMed

    Islam, Md Shahidul; Rahaman, Md Saifur; Yeum, Jeong Hyun

    2015-01-22

    A novel super-absorbent material was fabricated by electrospinning the natural polysaccharide pullulan (PULL) with polyvinyl alcohol (PVA) and montmorillonite (MMT) clay to form nonwoven webs, which were then heat treated. Transmission electron microscopy (TEM) micrographs, X-ray diffraction (XRD) patterns, and Fourier transform infrared (FTIR) analysis of the novel super-absorbent nanofibers suggest the coexistence of PULL, PVA, and MMT through the exfoliation of MMT layers in the super-absorbent nanofiber composite. The heat-treated PULL/PVA/MMT webs loaded with 5 wt% MMT electrospun nanofibers exhibited a water absorbency of 143.42 g g(-1) in distilled water and a water absorbency of 39.75 g g(-1) in a 0.9 wt% NaCl solution. Under extremely dry conditions, the PULL/PVA/MMT webs exhibited the ability to retain 43% distilled water and 38% saline water after being exposed to the atmosphere for one week. The heat treatment improved the crystallinity of the electrospun PULL/PVA/MMT super-absorbent webs and thus made the webs highly stable in aqueous environments. Overall, the addition of MMT resulted in improved thermal stability and mechanical properties and increased the water absorbency of the PULL/PVA/MMT composite. PMID:25439870

  2. Bioenhanced oral curcumin nanoparticles: Role of carbohydrates.

    PubMed

    D'Souza, Anisha A; Devarajan, Padma V

    2016-01-20

    The paper discusses polysaccharide-adsorbed curcumin-Gantrez nanoparticles for bioenhancement of oral curcumin. Nanoparticles revealed no change in size over time in pH 1.2 and 7.4, and a rapid drug release in pH 1.2 and 7.4 medium containing surfactant. Without adsorbed polysaccharides, nanoparticles exhibited high Cmax (61.3 ± 22.3 ng/mL), sustained plasma concentration up to 24h and 117% absolute bioavailability, attributed to bioadhesion. In contrast galactose polysaccharides arabinogalactan and kappa-carrageenan adsorbed nanoparticles exhibited rapid absorption with higher Cmax of 109.5 ± 31.2 ng/mL and 92.3 ± 21.2 ng/mL, respectively, but faster elimination and absolute bioavailability of greater than 25%. The glucose polysaccharide pullulan adsorbed nanoparticles exhibited significantly lower Cmax (39.7 ± 20.6 ng/mL) and bioavailability (13%). Lower bioavailability of polysaccharide adsorbed nanoparticles was attributed to high metabolism of curcumin in the intestine as a result of faster gastric elimination and high intestinal localization. However polysaccharide-adsorbed nanoparticles could play an important role in bioenhancement of drugs specifically those exhibiting good stability across the gastrointestinal tract.

  3. Carbohydrate Nanoparticles for Brain Delivery.

    PubMed

    Lalatsa, A; Barbu, E

    2016-01-01

    Many brain tumors and neurological diseases can greatly benefit from the use of emerging nanotechnologies based on targeted nanomedicines that are able to noninvasively transport highly potent and specific pharmaceuticals across the blood-brain barrier. Carbohydrates have received considerable interest as materials for drug carriers due to their natural origin and inherent biodegradability and biocompatibility, as well as due to their hydrophilic character and ease of chemical modification combined with low cost and the possibility for large-scale manufacturing. This chapter provides an overview of the latest research involving the use of carbohydrate-based nanoparticles for drug delivery to the central nervous system. After reviewing the challenges posed by delivering drugs into the brain, the current state-of-the-art approaches for delivery of actives across the blood-brain barrier, including invasive and noninvasive strategies, are presented. A particular focus has been placed on chitosan polymers as they are among the most promising carbohydrate nanocarriers for the preparation and testing of chitosan-based nanomedicines that led, in preclinical proof-of-concept studies, to enhanced brain drug levels and increased pharmacodynamics responses after intravenous, nasal, and oral administration. While chitosan nanoparticles are to date among the most studied and most promising carriers, approaches based on other polysaccharides such as dextran, pullulan, and cellulose warrant further research in the attempt to advance the existing technologies for overcoming the blood-brain barrier. PMID:27678176

  4. Nonclinical Evaluation of Novel Cationically Modified Polysaccharide Antidotes for Unfractionated Heparin

    PubMed Central

    Kalaska, Bartlomiej; Kaminski, Kamil; Sokolowska, Emilia; Czaplicki, Dominik; Kujdowicz, Monika; Stalinska, Krystyna; Bereta, Joanna; Szczubialka, Krzysztof; Pawlak, Dariusz; Nowakowska, Maria; Mogielnicki, Andrzej

    2015-01-01

    Protamine, the only registered antidote of unfractionated heparin (UFH), may produce a number of adverse effects, such as anaphylactic shock or serious hypotension. We aimed to develop an alternative UFH antidote as efficient as protamine, but safer and easier to produce. As a starting material, we have chosen generally non-toxic, biocompatible, widely available, inexpensive, and easy to functionalize polysaccharides. Our approach was to synthesize, purify and characterize cationic derivatives of dextran, hydroxypropylcellulose, pullulan and γ-cyclodextrin, then to screen them for potential heparin-reversal activity using an in vitro assay and finally examine efficacy and safety of the most active polymers in Wistar rat and BALB/c mouse models of experimentally induced arterial and venous thrombosis. Efficacy studies included the measurement of thrombus formation, activated partial thromboplastin time, bleeding time, and anti-factor Xa activity; safety studies included the measurement of hemodynamic, hematologic and immunologic parameters. Linear, high molecular weight dextran substituted with glycidyltrimethylammonium chloride groups at a ratio of 0.65 per glucose unit (Dex40-GTMAC3) is the most potent and the safest UFH inhibitor showing activity comparable to that of protamine while possessing lower immunogenicity. Cationic polysaccharides of various structures neutralize UFH. Dex40-GTMAC3 is a promising and potentially better UFH antidote than protamine. PMID:25781030

  5. The crosslinking of polysaccharides with polyamines and dextran-polyallylamine antibacterial hydrogels.

    PubMed

    O'Connor, Naphtali A; Abugharbieh, Ahmad; Yasmeen, Farzana; Buabeng, Emmanuel; Mathew, Steve; Samaroo, Diana; Cheng, Hai-Ping

    2015-01-01

    A facile modular approach to rapidly prepare pH-responsive hydrogels by crosslinking polysaccharides with polyamines is demonstrated. Hydrogels are prepared by first reacting the less reactive polysaccharides with the cross-linker epichlorohydrin and completed by the addition of polyamines. The crosslinking of polysaccharides with polyamines provides a facile method for incorporating functionality into polysaccharide based hydrogels. This process is demonstrated with the polysaccharides dextran, pullulan and carboxymethyl cellulose and with the polyamines polyallylamine and polyethylene imine. The hydrogels were characterized by FTIR and swelling studies, which showed pH-dependent swelling due to the presence of the polyamine. The hydrogels can also be tailored by varying the mass ratio between the polysaccharide and polyamine. Absorption studies of organic analytes showed the polyamine content affecting the uptake of a charged substrate (methylene blue) and no effect on a neutral substrate (6-methyl coumarin). This synthetic method was also used to prepare hydrogels with antibacterial activity against E. coli and S. aureus by utilizing an amphiphilic polyallylamine. PMID:25128095

  6. Rhizopus microsporus var. rhizopodiformis: a thermotolerant fungus with potential for production of thermostable amylases.

    PubMed

    Peixoto, Simone C; Jorge, João A; Terenzi, Héctor F; Polizeli, Maria de Lourdes T M

    2003-12-01

    The effect of several nutritional and environmental parameters on growth and amylase production from Rhizopus microsporus var. rhizopodiformis was analysed. This fungus was isolated from soil of the Brazilian "cerrado" and produced high levels of amylolytic activity at 45 degrees C in liquid medium supplemented with starch, sugar cane bagasse, oat meal or cassava flour. Glucose in the culture medium drastically repressed the amylolytic activity. The products of hydrolysis were analysed by thin layer chromatography, and glucose was detected as the main component. The amylolytic activity hydrolysed several substrates, such as amylopectin, amylase, glycogen, pullulan, starch, and maltose. Glucose was always the main end product detected by high-pressure liquid chromatography analysis. These results indicated that the amylolytic activity studied is a glucoamylase, but there were also low levels of alpha-amylase. As compared to other fungi, R. microsporus var. rhizopodiformis can be considered an efficient producer of thermostable amylases, using raw residues of low cost as substrates. This information is of technological value, considering the importance of amylases for industrial hydrolysis.

  7. Development of a Teat Bio-sealant and Evaluation of its Technological and Functional Properties.

    PubMed

    Serna-Cock, Liliana; Pabón-Rodríguez, Omar Vladimir

    2016-06-01

    A teat bio-sealant was developed using Weissella cibaria, and the bio-sealant's technological and functional properties were assessed. The development included four experimental phases that were analyzed using independent experimental designs. Initially, sterilized or pasteurized Aloe vera gels were used, and the effect of heat treatment was investigated. In the second phase, the effects of time, storage temperature, and addition of cryopreservatives on the viability of the probiotic were observed. The third phase consisted of evaluating the synergistic effects of the cryopreservatives. The fourth phase involved selecting a material that would provide viscosity to the teat sealant. Technological and functional properties were measured in terms of viability of W. cibaria, and antimicrobial activity against Staphylococcus aureus and Streptococcus agalactiae was also analyzed. A mixture of milk powder and glycerol preserved this antimicrobial activity. Pullulan provided greater viscosity and maintained the technological and functional properties of the bio-sealant for 29 days. This teat bio-sealant can be used as an alternative for the prevention of bovine mastitis. PMID:27084703

  8. Effect of C-terminal truncation on enzyme properties of recombinant amylopullulanase from Thermoanaerobacter pseudoethanolicus.

    PubMed

    Lin, Fu-Pang; Ho, Yi-Hsuan; Lin, Hsu-Yang; Lin, Hui-Ju

    2012-05-01

    The smallest and enzymatically active molecule, TetApuQ818, was localized within the C-terminal Q818 amino acid residue after serial C-terminal truncation analysis of the recombinant amylopullulanase molecule (TetApuM955) from Thermoanaerobacter pseudoethanolicus. Kinetic analyses indicated that the overall catalytic efficiency, k (cat)/K (m), of TetApuQ818 was 8-32% decreased for the pullulan and the soluble starch substrate, respectively. Changes to the substrate affinity, K (m), and the turnover rate, k (cat), were decreased significantly in both enzymatic activities of TetApuQ818. TetApuQ818 exhibited less thermostability than TetApuM955 when the temperature was raised above 85°C, but it had similar substrate-binding ability and hydrolysis products toward various substrates as TetApuM955 did. Both enzymes showed similar spectroscopies of fluorescence and circular dichroism, suggesting the active folding conformation was maintained after this C-terminal Q818 deletion. This study suggested that the binding ability of insoluble starch by TetApuM955 did not rely on the putative C-terminal carbohydrate binding module family 20 (CBM20) and two FnIII regions of TetApu, though the integrity of the AamyC module of TetApuQ818 was required for the enzyme activity. PMID:22392283

  9. Gluconic acid production.

    PubMed

    Anastassiadis, Savas; Morgunov, Igor G

    2007-01-01

    Gluconic acid, the oxidation product of glucose, is a mild neither caustic nor corrosive, non toxic and readily biodegradable organic acid of great interest for many applications. As a multifunctional carbonic acid belonging to the bulk chemicals and due to its physiological and chemical characteristics, gluconic acid itself, its salts (e.g. alkali metal salts, in especially sodium gluconate) and the gluconolactone form have found extensively versatile uses in the chemical, pharmaceutical, food, construction and other industries. Present review article presents the comprehensive information of patent bibliography for the production of gluconic acid and compares the advantages and disadvantages of known processes. Numerous manufacturing processes are described in the international bibliography and patent literature of the last 100 years for the production of gluconic acid from glucose, including chemical and electrochemical catalysis, enzymatic biocatalysis by free or immobilized enzymes in specialized enzyme bioreactors as well as discontinuous and continuous fermentation processes using free growing or immobilized cells of various microorganisms, including bacteria, yeast-like fungi and fungi. Alternatively, new superior fermentation processes have been developed and extensively described for the continuous and discontinuous production of gluconic acid by isolated strains of yeast-like mold Aureobasidium pullulans, offering numerous advantages over the traditional discontinuous fungi processes.

  10. Structures of wall heterogalactomannans isolated from three genera of entomopathogenic fungi.

    PubMed

    Bernabé, Manuel; Salvachúa, Davinia; Jiménez-Barbero, Jesús; Leal, Juan Antonio; Prieto, Alicia

    2011-09-01

    O-linked heterogalactomannans with similar structural features have been purified from the fungal walls of the entomopathogenic fungi Lecanicillium muscarium, Beauveria bassiana, Beauveriabrongniartii, and Cordyceps sphingum. Their composition and structure have been determined using acid hydrolysis, methylation analysis, gas-liquid chromatography-mass spectrometry (GC-MS) and Nuclear magnetic resonance spectroscopy (NMR). All structures have an α-(1→6)-mannose backbone, but one of the two strains of L. muscarium included in this study contained an acidic heterogalactomannan instead of the neutral polysaccharide isolated in the rest of the species analyzed. Sequence analysis of the internal transcribed spacer (ITS) region of this strain indicated that it belongs to the related genus Simplicillium, displaying low identity (83%) with the closest Lecanicillium species. This is a new demonstration of the structural diversity of fungal wall heteromannans and validates their interest as chemotaxonomic markers. The production of a pullulan-like extracellular polysaccharide in strain CBS 413.70C of L. muscarium is also reported. PMID:21872183

  11. Heat stable antimicrobial activity of Allium ascalonicum against bacteria and fungi.

    PubMed

    Amin, M; Kapadnis, B P

    2005-08-01

    To study antimicrobial activity of shallot in comparison with that of garlic and onion against 23 strains of fungi and bacteria, water extracts of garlic, shallot and onion bulbs were prepared. Each extract was studied in different forms for their antimicrobial activity viz., fresh extract, dry extract and autoclaved extract. Minimal inhibitory concentration and minimal lethal concentrations of these extracts were determined against all organisms by broth dilution susceptibility test. Fresh extract of garlic showed greater antimicrobial activity as compared to similar extracts of onion and shallot. However, dried and autoclaved extracts of shallot showed more activity than similar extracts of onion and garlic. Fungi were more sensitive to shallot extract than bacteria. Amongst bacteria, B. cereus was most sensitive (MIC=5 mg ml(-1)). The lowest minimum bactericidal concentration of shallot extract amongst bacteria tested was 5 mg ml(-1) for B. cereus. Amongst fungi, Aureobasidium pullulans and Microsporum gypseum were most sensitive (MIC= 0.15 mg ml(-1)). The lowest minimum lethal concentration was 2.5 mg ml(-1) for Microsporum gypseum and Trichophyton mentagrophytes. It was therefore, expected that the antimicrobial principle of shallot was different than the antimicrobial compounds of onion and garlic. In addition, the antimicrobial component of the shallot extract was stable at 121 degrees C.

  12. Characterization of a pH and detergent-tolerant, cold-adapted type I pullulanase from Exiguobacterium sp. SH3.

    PubMed

    Rajaei, Sarah; Noghabi, Kambiz Akbari; Sadeghizadeh, Majid; Zahiri, Hossein Shahbani

    2015-11-01

    A pullulanase-encoding gene from psychrotrophic Exiguobacterium sp. SH3 was cloned and expressed in both E. coli and Bacillus subtilis. The functional recombinant enzyme (Pul-SH3) was purified as a His-tagged protein. Pul-SH3 was characterized to be a cold-adapted type I pullulanase with maximum activity at 45 °C. Using fluorescence spectroscopy, the melting temperature of Pul-SH3 was determined to be about 52 °C. The enzyme was able to hydrolyze pullulan, soluble starch, potato starch, and rice flour. It was exceptionally tolerant in the pH range of 4-11, exhibiting maximum activity at pH 8.5 and more than 60% of the activity in the pH range of 5-11. Being a detergent-tolerant pullulanase, Pul-SH3 retained 99, 89, and 54% of its activity at 10% concentration of Triton-X100, Tween 20, and SDS, respectively. The enzyme also exhibited an activity of 80.4 and 93.7% in the presence of two commercial detergents, Rika (7.5% v/v) and Fadisheh (2.5% w/v), respectively. The enzyme was even able to remain active by 54.5 and 85% after 10-day holding with the commercial detergents. Thermal stability of the enzyme could w on silica. Pul-SH3 with several industrially important characteristics seems desirable for cold hydrolysis of starch. PMID:26349928

  13. Different drying technologies and alternation of mycobiots in the raw material of Hyssopus officinalis L.

    PubMed

    Raila, Algirdas; Lugauskas, Albinas; Kemzūraite, Aurelija; Zvicevicius, Egidijus; Ragazinskiene, Ona; Railiene, Marija

    2009-01-01

    Contamination of medicinal plant mass with mycobiots is one of the negative factors deteriorating the quality of raw material. In order to evaluate the impact of the yield processing technologies upon the changes of mycobiots in raw material, the mycobiotic conditions of herb hyssop (Hyssopus officinalis L.) raw material were evaluated under various regimes of active ventilation and optimization of the drying parameters. The impact of ventilation intensity and temperature of drying agent upon the changes and abundance of mycobiota species in medicinal raw material was determined. Irrespective of the temperature of the airflow, the strongest suppressive effect upon the mycobiotic contamination in Hyssopi herba was produced by the 5,000 m3 x (t x h)(-1) airflow. Analysis of the isolated fungi revealed the prevalence of Penicillium, Aspergillus, Alternaria, Cladosporium, Mucor, Rhizopus species in the raw material. In separate samples Botrytis cinerea, Sclerotinia sclerotiorum, Aureobasidium pullulans, Chrysosporium merdarium, Cladorrhinum foecundissimum, Ulocladium consortiale, Trichoderma hamatum, T. harzianum, Gilmaniella humicola, Talaromyces flavus, Rhizomucor pusillus, Hansfordia ovalispora, Verticicladium trifi dum, Trichosporiella cerebriformis micromycetes were also rather abundant. Detection of the above-mentioned micromycetes in herb hyssop samples differed, and partially depended upon the medium used for their isolation. PMID:19630202

  14. Fungi associated with black mould on baobab trees in southern Africa.

    PubMed

    Cruywagen, Elsie M; Crous, Pedro W; Roux, Jolanda; Slippers, Bernard; Wingfield, Michael J

    2015-07-01

    There have been numerous reports in the scientific and popular literature suggesting that African baobab (Adansonia digitata) trees are dying, with symptoms including a black mould on their bark. The aim of this study was to determine the identity of the fungi causing this black mould and to consider whether they might be affecting the health of trees. The fungi were identified by sequencing directly from mycelium on the infected tissue as well as from cultures on agar. Sequence data for the ITS region of the rDNA resulted in the identification of four fungi including Aureobasidium pullulans, Toxicocladosporium irritans and a new species of Rachicladosporium described here as Rachicladosporium africanum. A single isolate of an unknown Cladosporium sp. was also found. These fungi, referred to here as black mould, are not true sooty mould fungi and they were shown to penetrate below the bark of infected tissue, causing a distinct host reaction. Although infections can lead to dieback of small twigs on severely infected branches, the mould was not found to kill trees.

  15. Status, Antimicrobial Mechanism, and Regulation of Natural Preservatives in Livestock Food Systems.

    PubMed

    Lee, Na-Kyoung; Paik, Hyun-Dong

    2016-01-01

    This review discusses the status, antimicrobial mechanisms, application, and regulation of natural preservatives in livestock food systems. Conventional preservatives are synthetic chemical substances including nitrates/nitrites, sulfites, sodium benzoate, propyl gallate, and potassium sorbate. The use of artificial preservatives is being reconsidered because of concerns relating to headache, allergies, and cancer. As the demand for biopreservation in food systems has increased, new natural antimicrobial compounds of various origins are being developed, including plant-derived products (polyphenolics, essential oils, plant antimicrobial peptides (pAMPs)), animal-derived products (lysozymes, lactoperoxidase, lactoferrin, ovotransferrin, antimicrobial peptide (AMP), chitosan and others), and microbial metabolites (nisin, natamycin, pullulan, ε-polylysine, organic acid, and others). These natural preservatives act by inhibiting microbial cell walls/membranes, DNA/RNA replication and transcription, protein synthesis, and metabolism. Natural preservatives have been recognized for their safety; however, these substances can influence color, smell, and toxicity in large amounts while being effective as a food preservative. Therefore, to evaluate the safety and toxicity of natural preservatives, various trials including combinations of other substances or different food preservation systems, and capsulation have been performed. Natamycin and nisin are currently the only natural preservatives being regulated, and other natural preservatives will have to be legally regulated before their widespread use.

  16. Status, Antimicrobial Mechanism, and Regulation of Natural Preservatives in Livestock Food Systems.

    PubMed

    Lee, Na-Kyoung; Paik, Hyun-Dong

    2016-01-01

    This review discusses the status, antimicrobial mechanisms, application, and regulation of natural preservatives in livestock food systems. Conventional preservatives are synthetic chemical substances including nitrates/nitrites, sulfites, sodium benzoate, propyl gallate, and potassium sorbate. The use of artificial preservatives is being reconsidered because of concerns relating to headache, allergies, and cancer. As the demand for biopreservation in food systems has increased, new natural antimicrobial compounds of various origins are being developed, including plant-derived products (polyphenolics, essential oils, plant antimicrobial peptides (pAMPs)), animal-derived products (lysozymes, lactoperoxidase, lactoferrin, ovotransferrin, antimicrobial peptide (AMP), chitosan and others), and microbial metabolites (nisin, natamycin, pullulan, ε-polylysine, organic acid, and others). These natural preservatives act by inhibiting microbial cell walls/membranes, DNA/RNA replication and transcription, protein synthesis, and metabolism. Natural preservatives have been recognized for their safety; however, these substances can influence color, smell, and toxicity in large amounts while being effective as a food preservative. Therefore, to evaluate the safety and toxicity of natural preservatives, various trials including combinations of other substances or different food preservation systems, and capsulation have been performed. Natamycin and nisin are currently the only natural preservatives being regulated, and other natural preservatives will have to be legally regulated before their widespread use. PMID:27621697

  17. Membrane-bound amylopullulanase is essential for starch metabolism of Sulfolobus acidocaldarius DSM639.

    PubMed

    Choi, Kyoung-Hwa; Cha, Jaeho

    2015-09-01

    Sulfolobus acidocaldarius DSM639 produced an acid-resistant membrane-bound amylopullulanase (Apu) during growth on starch as a sole carbon and energy source. The physiological role of Apu in starch metabolism was investigated by the growth and starch degradation pattern of apu disruption mutant as well as biochemical properties of recombinant Apu. The Δapu mutant lost the ability to grow in minimal medium in the presence of starch, and the amylolytic activity observed in the membrane fraction of the wild-type strain was not detected in the Δapu mutant when the cells were grown in YT medium. The purified membrane-bound Apu initially hydrolyzed starch, amylopectin, and pullulan into various sizes of maltooligosaccharides, and then produced glucose, maltose, and maltotriose in the end, indicating Apu is a typical endo-acting glycoside hydrolase family 57 (GH57) amylopullulanase. The maltose and maltotriose observed in the culture medium during the exponential and stationary phase growth indicates that Apu is the essential enzyme to initially hydrolyze the starch into small maltooligosaccharides to be transported into the cell.

  18. Characterization of recombinant amylopullulanase (gt-apu) and truncated amylopullulanase (gt-apuT) of the extreme thermophile Geobacillus thermoleovorans NP33 and their action in starch saccharification.

    PubMed

    Nisha, M; Satyanarayana, T

    2013-07-01

    A gene encoding amylopullulanase (gt-apu) of the extremely thermophilic Geobacillus thermoleovorans NP33 was cloned and expressed in Escherichia coli. The gene has an open reading frame of 4,965 bp that encodes a protein of 1,655 amino acids with molecular mass of 182 kDa. The six conserved regions, characteristic of GH13 family, have been detected in gt-apu. The recombinant enzyme has only one active site for α-amylase and pullulanase activities based on the enzyme kinetic analyses in a system that contains starch as well as pullulan as competing substrates and response to inhibitors. The end-product analysis confirmed that this is an endoacting enzyme. The specific enzyme activities for α-amylase and pullulanase of the truncated amylopullulanase (gt-apuT) are higher than gt-apu. Both enzymes exhibited similar temperature (60 °C) and pH (7.0) optima, although gt-apuT possessed a higher thermostability than gt-apu. The overall catalytic efficiency (K(cat)/K(m)) of gt-apuT is greater than that of gt-apu, with almost similar substrate specificities. The C-terminal region of gt-apu appeared to be non-essential, and furthermore, it negatively affects the substrate binding and stability of the enzyme.

  19. Carbohydrate Polymers for Nonviral Nucleic Acid Delivery

    PubMed Central

    Sizovs, Antons; McLendon, Patrick M.; Srinivasachari, Sathya

    2014-01-01

    Carbohydrates have been investigated and developed as delivery vehicles for shuttling nucleic acids into cells. In this review, we present the state of the art in carbohydrate-based polymeric vehicles for nucleic acid delivery, with the focus on the recent successes in preclinical models, both in vitro and in vivo. Polymeric scaffolds based on the natural polysaccharides chitosan, hyaluronan, pullulan, dextran, and schizophyllan each have unique properties and potential for modification, and these results are discussed with the focus on facile synthetic routes and favorable performance in biological systems. Many of these carbohydrates have been used to develop alternative types of biomaterials for nucleic acid delivery to typical polyplexes, and these novel materials are discussed. Also presented are polymeric vehicles that incorporate copolymerized carbohydrates into polymer backbones based on polyethylenimine and polylysine and their effect on transfection and biocompatibility. Unique scaffolds, such as clusters and polymers based on cyclodextrin (CD), are also discussed, with the focus on recent successes in vivo and in the clinic. These results are presented with the emphasis on the role of carbohydrate and charge on transfection. Use of carbohydrates as molecular recognition ligands for cell-type specific delivery is also briefly reviewed. We contend that carbohydrates have contributed significantly to progress in the field of non-viral DNA delivery, and these new discoveries are impactful for developing new vehicles and materials for treatment of human disease. PMID:21504102

  20. Asialoglycoprotein receptor mediated hepatocyte targeting - strategies and applications.

    PubMed

    D'Souza, Anisha A; Devarajan, Padma V

    2015-04-10

    Hepatocyte resident afflictions continue to affect the human population unabated. The asialoglycoprotein receptor (ASGPR) is primarily expressed on hepatocytes and minimally on extra-hepatic cells. This makes it specifically attractive for receptor-mediated drug delivery with minimum concerns of toxicity. ASGPR facilitates internalization by clathrin-mediated endocytosis and exhibits high affinity for carbohydrates specifically galactose, N-acetylgalactosamine and glucose. Isomeric forms of sugar, galactose density and branching, spatial geometry and galactose linkages are key factors influencing ligand-receptor binding. Popular ligands for ASGPR mediated targeting are carbohydrate polymers, arabinogalactan and pullulan. Other ligands include galactose-bearing glycoproteins, glycopeptides and galactose modified polymers and lipids. Drug-ligand conjugates provide a viable strategy; nevertheless ligand-anchored nanocarriers provide an attractive option for ASGPR targeted delivery and are widely explored. The present review details various ligands and nanocarriers exploited for ASGPR mediated delivery of drugs to hepatocytes. Nanocarrier properties affecting ASGPR mediated uptake are discussed at length. The review also highlights the clinical relevance of ASGPR mediated targeting and applications in diagnostics. ASGPR mediated hepatocyte targeting provides great promise for improved therapy of hepatic afflictions.

  1. Electrospun novel super-absorbent based on polysaccharide-polyvinyl alcohol-montmorillonite clay nanocomposites.

    PubMed

    Islam, Md Shahidul; Rahaman, Md Saifur; Yeum, Jeong Hyun

    2015-01-22

    A novel super-absorbent material was fabricated by electrospinning the natural polysaccharide pullulan (PULL) with polyvinyl alcohol (PVA) and montmorillonite (MMT) clay to form nonwoven webs, which were then heat treated. Transmission electron microscopy (TEM) micrographs, X-ray diffraction (XRD) patterns, and Fourier transform infrared (FTIR) analysis of the novel super-absorbent nanofibers suggest the coexistence of PULL, PVA, and MMT through the exfoliation of MMT layers in the super-absorbent nanofiber composite. The heat-treated PULL/PVA/MMT webs loaded with 5 wt% MMT electrospun nanofibers exhibited a water absorbency of 143.42 g g(-1) in distilled water and a water absorbency of 39.75 g g(-1) in a 0.9 wt% NaCl solution. Under extremely dry conditions, the PULL/PVA/MMT webs exhibited the ability to retain 43% distilled water and 38% saline water after being exposed to the atmosphere for one week. The heat treatment improved the crystallinity of the electrospun PULL/PVA/MMT super-absorbent webs and thus made the webs highly stable in aqueous environments. Overall, the addition of MMT resulted in improved thermal stability and mechanical properties and increased the water absorbency of the PULL/PVA/MMT composite.

  2. MAPLE activities and applications in gas sensors

    NASA Astrophysics Data System (ADS)

    Jelínek, Miroslav; Remsa, Jan; Kocourek, Tomáš; Kubešová, Barbara; Schůrek, Jakub; Myslík, Vladimír

    2011-11-01

    During the last decade, many groups have grown thin films of various organic materials by the cryogenic Matrix Assisted Pulsed Laser Evaporation (MAPLE) technique with a wide range of applications. This contribution is focused on the summary of our results with deposition and characterization of thin films of fibrinogen, pullulan derivates, azo-polyurethane, cryoglobulin, polyvinyl alcohol, and bovine serum albumin dissolved in physiological serum, dimethyl sulfoxide, sanguine plasma, phosphate buffer solution, H2O, ethylene glycol, and tert-butanol. MAPLE films were characterized using FTIR, AFM, Raman scattering, and SEM. For deposition, a special hardware was developed including a unique liquid nitrogen cooled target holder. Overview of MAPLE thin film applications is given. We studied SnAcAc, InAcAc, SnO2, porphyrins, and polypyrrole MAPLE fabricated films as small resistive gas sensors. Sensors were tested with ozone, nitrogen dioxide, hydrogen, and water vapor gases. In the last years, our focus was on the study of fibrinogen-based scaffolds for application in tissue engineering, wound healing, and also as a part of layers for medical devices.

  3. Characterization of pectinase activity for enology from yeasts occurring in Argentine Bonarda grape.

    PubMed

    Merín, María Gabriela; Martín, María Carolina; Rantsiou, Kalliopi; Cocolin, Luca; de Ambrosini, Vilma Inés Morata

    2015-01-01

    Pectinolytic enzymes are greatly important in winemaking due to their ability to degrade pectic polymers from grape, contributing to enhance process efficiency and wine quality. This study aimed to analyze the occurrence of pectinolytic yeasts during spontaneous fermentation of Argentine Bonarda grape, to select yeasts that produce extracellular pectinases and to characterize their pectinolytic activity under wine-like conditions. Isolated yeasts were grouped using PCR-DGGE and identified by partial sequencing of 26S rRNA gene. Isolates comprised 7 genera, with Aureobasidium pullulans as the most predominant pectinolytic species, followed by Rhodotorula dairenensis and Cryptococcus saitoi. No pectinolytic activity was detected among ascomycetous yeasts isolated on grapes and during fermentation, suggesting a low occurrence of pectinolytic yeast species in wine fermentation ecosystem. This is the first study reporting R. dairenensis and Cr. saitoi species with pectinolytic activity. R. dairenensis GM-15 produced pectinases that proved to be highly active at grape pH, at 12 °C, and under ethanol and SO2 concentrations usually found in vinifications (pectinase activity around 1.1 U/mL). This strain also produced cellulase activity at 12 °C and pH 3.5, but did not produce β-glucosidase activity under these conditions. The strain showed encouraging enological properties for its potential use in low-temperature winemaking. PMID:26413065

  4. In vitro inhibition of postharvest pathogens of fruit and control of gray mold of strawberry and green mold of citrus by aureobasidin A.

    PubMed

    Liu, Xiaoping; Wang, Jiye; Gou, Ping; Mao, Cungui; Zhu, Zeng-Rong; Li, Hongye

    2007-11-01

    Aureobasidin A (AbA), an antifungal cyclic depsipeptide antibiotic produced by Aureobasidium pullulans R106, has previously been shown to be effective against a wide range of fungi and protozoa. Here we report the inhibitory effects of AbA on spore germination, germ tuber elongation and hyphal growth of five pathogenic fungi including Penicillium digitatum, P. italicum, P. expansum, Botrytis cinerea and Monilinia fructicola, which are major pathogens causing postharvest diseases of a variety of fruits. AbA inhibited five pathogenic fungi by reducing conidial germination rates, delaying conidial germination initiation, restricting elongation of germ tuber and mycelium, as well as inducing abnormal alternations of morphology of germ tubes and hyphae of these fungi. The sensitivity of these fungi to AbA was pathogen species-dependent. P. digitatum was the most sensitive and M. fructicola the least. Importantly, AbA at 50 microg/ml was effective in controlling the citrus green mold and in reducing the strawberry gray mold incidence and severity, caused by P. digitatum and B. cinerea, respectively, after artificial inoculation. AbA and/or its analogs, therefore, hold promise as relatively safe and promising fungicide candidates to control postharvest decays of fruits, because AbA targets the inositol phosphorylceramide (IPC) synthase, an enzyme essential for fungi but absent from mammals.

  5. Transposon Mutagenesis in Bifidobacterium breve: Construction and Characterization of a Tn5 Transposon Mutant Library for Bifidobacterium breve UCC2003

    PubMed Central

    Ruiz, Lorena; Motherway, Mary O’Connell; Lanigan, Noreen; van Sinderen, Douwe

    2013-01-01

    Bifidobacteria are claimed to contribute positively to human health through a range of beneficial or probiotic activities, including amelioration of gastrointestinal and metabolic disorders, and therefore this particular group of gastrointestinal commensals has enjoyed increasing industrial and scientific attention in recent years. However, the molecular mechanisms underlying these probiotic mechanisms are still largely unknown, mainly due to the fact that molecular tools for bifidobacteria are rather poorly developed, with many strains lacking genetic accessibility. In this work, we describe the generation of transposon insertion mutants in two bifidobacterial strains, B. breve UCC2003 and B. breve NCFB2258. We also report the creation of the first transposon mutant library in a bifidobacterial strain, employing B. breve UCC2003 and a Tn5-based transposome strategy. The library was found to be composed of clones containing single transposon insertions which appear to be randomly distributed along the genome. The usefulness of the library to perform phenotypic screenings was confirmed through identification and analysis of mutants defective in D-galactose, D-lactose or pullulan utilization abilities. PMID:23737995

  6. Cellulose acetate based 3-dimensional electrospun scaffolds for skin tissue engineering applications.

    PubMed

    Atila, Deniz; Keskin, Dilek; Tezcaner, Ayşen

    2015-11-20

    Skin defects that are not able to regenerate by themselves are among the major problems faced. Tissue engineering approach holds promise for treating such defects. Development of tissue-mimicking-scaffolds that can promote healing process receives an increasing interest in recent years. In this study, 3-dimensional electrospun cellulose acetate (CA) pullulan (PULL) scaffolds were developed for the first time. PULL was intentionally used to obtain 3D structures with adjustable height. It was removed from the electrospun mesh to increase the porosity and biostability. Different ratios of the polymers were electrospun and analyzed with respect to degradation, porosity, and mechanical properties. It has been observed that fiber diameter, thickness and porosity of scaffolds increased with increased PULL content, on the other hand this resulted with higher degradation of scaffolds. Mechanical strength of scaffolds was improved after PULL removal suggesting their suitability as cell carriers. Cell culture studies were performed with the selected scaffold group (CA/PULL: 50/50) using mouse fibroblastic cell line (L929). In vitro cell culture tests showed that cells adhered, proliferated and populated CA/PULL (50/50) scaffolds showing that they are cytocompatible. Results suggest that uncrosslinked CA/PULL (50/50) electrospun scaffolds hold potential for skin tissue engineering applications. PMID:26344279

  7. Molecular identification of fungal pathogens in nodular skin lesions of cats.

    PubMed

    Bernhardt, Anne; von Bomhard, Wolf; Antweiler, Elisabeth; Tintelnot, Kathrin

    2015-02-01

    In a retrospective study, we investigated 52 formalin-fixed, paraffin-embedded (FFPE) samples from cats with histologically confirmed cutaneous and subcutaneous mycoses to determine if the pathogens could be identified by molecular methods. Aim of the study was to obtain a deep understanding of the spectrum of infectious agents, which, as we hypothesized, was not available by histopathology alone. Detection of feline and fungal DNA was achieved in 92.3% and 94.2% of the samples, respectively. Most of the subcutaneous infections in cats were caused by Alternaria spp. (63.5%), followed by Cryptococcus neoformans (7.7%), Histoplasma capsulatum (5.8%), Sporothrix spp. (3.8%), Aspergillus vitricola, Aureobasidium pullulans, Exophiala attenuata, Fusarium oxysporum, Lecythophora cateniformis, Microsporum canis, and Phialophora sp. (1.9% each). The results from molecular identification indicate that correct identifications of the fungal pathogens by histology alone were rarely possible. The spectrum of fungal pathogens identified after DNA extraction from FFPE samples was much broader than that expected by classical histopathology. This was especially noted in alternariosis in that the micromorphological pattern in tissue was misleading and could be confused with that of cryptococcosis. Due to different susceptibilities to antifungal agents, it is important to arrive at a definitive diagnosis, which might be possible by examination of the fungus recovered in culture and/or molecular methods, in addition to the histopathologic techniques. PMID:25550386

  8. Polysaccharides as cell carriers for tissue engineering: the use of cellulose in vascular wall reconstruction.

    PubMed

    Bačáková, L; Novotná, K; Pařízek, M

    2014-01-01

    Polysaccharides are long carbohydrate molecules of monosaccharide units joined together by glycosidic bonds. These biological polymers have emerged as promising materials for tissue engineering due to their biocompatibility, mostly good availability and tailorable properties. This complex group of biomolecules can be classified using several criteria, such as chemical composition (homo- and heteropolysaccharides), structure (linear and branched), function in the organism (structural, storage and secreted polysaccharides), or source (animals, plants, microorganisms). Polysaccharides most widely used in tissue engineering include starch, cellulose, chitosan, pectins, alginate, agar, dextran, pullulan, gellan, xanthan and glycosaminoglycans. Polysaccharides have been applied for engineering and regeneration of practically all tissues, though mostly at the experimental level. Polysaccharides have been tested for engineering of blood vessels, myocardium, heart valves, bone, articular and tracheal cartilage, intervertebral discs, menisci, skin, liver, skeletal muscle, neural tissue, urinary bladder, and also for encapsulation and delivery of pancreatic islets and ovarian follicles. For these purposes, polysaccharides have been applied in various forms, such as injectable hydrogels or porous and fibrous scaffolds, and often in combination with other natural or synthetic polymers or inorganic nanoparticles. The immune response evoked by polysaccharides is usually mild, and can be reduced by purifying the material or by choosing appropriate crosslinking agents.

  9. Characterization of pectinase activity for enology from yeasts occurring in Argentine Bonarda grape

    PubMed Central

    Merín, María Gabriela; Martín, María Carolina; Rantsiou, Kalliopi; Cocolin, Luca; de Ambrosini, Vilma Inés Morata

    2015-01-01

    Pectinolytic enzymes are greatly important in winemaking due to their ability to degrade pectic polymers from grape, contributing to enhance process efficiency and wine quality. This study aimed to analyze the occurrence of pectinolytic yeasts during spontaneous fermentation of Argentine Bonarda grape, to select yeasts that produce extracellular pectinases and to characterize their pectinolytic activity under wine-like conditions. Isolated yeasts were grouped using PCR-DGGE and identified by partial sequencing of 26S rRNA gene. Isolates comprised 7 genera, with Aureobasidium pullulans as the most predominant pectinolytic species, followed by Rhodotorula dairenensis and Cryptococcus saitoi. No pectinolytic activity was detected among ascomycetous yeasts isolated on grapes and during fermentation, suggesting a low occurrence of pectinolytic yeast species in wine fermentation ecosystem. This is the first study reporting R. dairenensis and Cr. saitoi species with pectinolytic activity. R. dairenensis GM-15 produced pectinases that proved to be highly active at grape pH, at 12 °C, and under ethanol and SO2 concentrations usually found in vinifications (pectinase activity around 1.1 U/mL). This strain also produced cellulase activity at 12 °C and pH 3.5, but did not produce β-glucosidase activity under these conditions. The strain showed encouraging enological properties for its potential use in low-temperature winemaking. PMID:26413065

  10. Evaluation of certain food additives and contaminants.

    PubMed

    2011-01-01

    This report represents the conclusions of a Joint FAO/WHO Expert Committee convened to evaluate the safety of various food additives, with a view to recommending acceptable daily intakes (ADIs) and to preparing specifications for identity and purity. The Committee also evaluated the risk posed by two food contaminants, with the aim of deriving tolerable intakes where appropriate and advising on risk management options for the purpose of public health protection. The first part of the report contains a general discussion of the principles governing the toxicological evaluation of and assessment of dietary exposure to food additives and contaminants. A summary follows of the Committee's evaluations of technical, toxicological and dietary exposure data for certain food additives (aluminium-containing food additives, Benzoe Tonkinensis, glycerol ester of gum rosin, glycerol ester of tall oil rosin, glycerol ester of wood rosin, octenyl succinic acid modified gum arabic, polydimethyl siloxane, Ponceau 4R, pullulan, pullulanase from Bacillus deromificans expressed in Bacillus licheniformis, Quinoline Yellow and Sunset Yellow FCF) and two food contaminants (cyanogenic glycosides and fumonisins). Specifications for the following food additives were revised: aluminium lakes of colouring matters; beta-apo-8'-carotenal; beta-apo-8'-carotenoic acid ethyl ester; beta-carotene, synthetic; hydroxypropyl methyl cellulose; magnesium silicate, synthetic; modified starches; nitrous oxide; sodium carboxymethyl cellulose; and sucrose monoesters of lauric, palmitic or stearic acid. Annexed to the report are tables summarizing the Committee's recommendations for dietary exposures to and toxicological evaluations of the food additives and contaminants considered.

  11. Development of a Teat Bio-sealant and Evaluation of its Technological and Functional Properties.

    PubMed

    Serna-Cock, Liliana; Pabón-Rodríguez, Omar Vladimir

    2016-06-01

    A teat bio-sealant was developed using Weissella cibaria, and the bio-sealant's technological and functional properties were assessed. The development included four experimental phases that were analyzed using independent experimental designs. Initially, sterilized or pasteurized Aloe vera gels were used, and the effect of heat treatment was investigated. In the second phase, the effects of time, storage temperature, and addition of cryopreservatives on the viability of the probiotic were observed. The third phase consisted of evaluating the synergistic effects of the cryopreservatives. The fourth phase involved selecting a material that would provide viscosity to the teat sealant. Technological and functional properties were measured in terms of viability of W. cibaria, and antimicrobial activity against Staphylococcus aureus and Streptococcus agalactiae was also analyzed. A mixture of milk powder and glycerol preserved this antimicrobial activity. Pullulan provided greater viscosity and maintained the technological and functional properties of the bio-sealant for 29 days. This teat bio-sealant can be used as an alternative for the prevention of bovine mastitis.

  12. The Structural Basis of Alpha-Glucan Recognition by a Family 41 Carbohydrate-Binding Module from Therotoga Maritima

    SciTech Connect

    van Bueren,A.; Boraston, A.

    2006-01-01

    Starch recognition by carbohydrate-binding modules (CBMs) is important for the activity of starch-degrading enzymes. The N-terminal family 41 CBM, TmCBM41 (from pullulanase PulA secreted by Thermotoga maritima) was shown to have {alpha}-glucan binding activity with specificity for {alpha}-1, 4-glucans but was able to tolerate the {alpha}-1, 6-linkages found roughly every three or four glucose units in pullulan. Using X-ray crystallography, the structures were solved for TmCBM41 in an uncomplexed form and in complex with maltotetraose and 63-{alpha}-d-glucosyl-maltotriose (GM3). Ligand binding was facilitated by stacking interactions between the {alpha}-faces of the glucose residues and two tryptophan side-chains in the two main subsites of the carbohydrate-binding site. Overall, this mode of starch binding is quite well conserved by other starch-binding modules. The structure in complex with GM3 revealed a third binding subsite with the flexibility to accommodate an {alpha}-1, 4- or an {alpha}-1, 6-linked glucose.

  13. Rapid bacterial degradation of polysaccharides in anoxic marine systems

    SciTech Connect

    Arnosti, C.; Repeta, D.J.; Blough, N.V. )

    1994-06-01

    Extracellular hydrolysis of organic macromolecules is often assumed to be the slow step in remineralization of organic matter. The authors tested this assumption by comparing the degradation of four polysaccharides (pullulan, laminarin, and two polysaccharides isolated from the marine cyanobacterium Synechococcus WH7335) to determine whether size, linkage position, or anomeric linkage affected rates or mechanisms of carbohydrate degradation by mixed cultures of anaerobic bacteria enriched from marine sediments. Gel permeation chromatography and nuclear magnetic resonance spectroscopy (NMR) were used to follow the extracellular conversion of high molecular weight polysaccharides to lower molecular weight polysaccharides and oligosaccharides which were subsequently remineralized. In all cases, substrate degradation was rapid. NMR spectra showed that preferential hydrolysis occurred at specific chemical linkages, and extracellular enzymatic hydrolysis of polysaccharides occurred far more rapidly than bacterial uptake and remineralization of the lower molecular weight oligosaccharides produced through enzymatic hydrolysis. Substrate size was not a significant determinant of remineralization rate: High molecular weight does not always correlate with slow degradation rate. The hypothesis that extracellular enzymatic hydrolysis is a slow step in the degradation of macromolecular organic matter in marine systems needs to be critically re-examined.

  14. Water quality and diversity of yeasts from tropical lakes and rivers from the Rio Doce basin in Southeastern Brazil

    PubMed Central

    Medeiros, Adriana O.; Missagia, Beatriz S.; Brandão, Luciana R.; Callisto, Marcos; Barbosa, Francisco A. R.; Rosa, Carlos A.

    2012-01-01

    Yeast communities were assessed in 14 rivers and four lakes from the Doce River basin in Brazil, during the rainy and dry seasons of the years 2000 and 2001. Water samples were collected at the subsurface in all sites. The following physical and chemical parameters were measured: temperature, dissolved oxygen, pH, electrical conductivity, total phosphorus, ortho-phosphate, ammonium, nitrate, nitrite and total nitrogen and the counts of faecal coliforms and heterotrophic bacteria were carried out to characterize the aquatic environmental sampled. The yeast counts were higher in aquatic environments with the highest counts of coliform and heterotrophic bacteria. These environments receive a high influx of domestic and industrial waste. A total of 317 isolates identified in forty eight yeast species were recorded in the sites sampled and the specie Aureobasidium pullulans were found in eleven out of eighteen sites sampled and some opportunistic pathogens such as the yeast species Candida krusei were isolated only in the polluted rivers with a positive correlation with the biotic and abiotic parameters that indicate sewage contamination. PMID:24031990

  15. Recombinant Cyclodextrinase from Thermococcus kodakarensis KOD1: Expression, Purification, and Enzymatic Characterization

    PubMed Central

    Lv, Xiaomin

    2015-01-01

    A gene encoding a cyclodextrinase from Thermococcus kodakarensis KOD1 (CDase-Tk) was identified and characterized. The gene encodes a protein of 656 amino acid residues with a molecular mass of 76.4 kDa harboring four conserved regions found in all members of the α-amylase family. A recombinant form of the enzyme was purified by ion-exchange chromatography, and its catalytic properties were examined. The enzyme was active in a broad range of pH conditions (pHs 4.0–10.0), with an optimal pH of 7.5 and a temperature optimum of 65°C. The purified enzyme preferred to hydrolyze β-cyclodextrin (CD) but not α- or γ-CD, soluble starch, or pullulan. The final product from β-CD was glucose. The Vmax and Km values were 3.13 ± 0.47 U mg−1 and 2.94 ± 0.16 mg mL−1 for β-CD. The unique characteristics of CDase-Tk with a low catalytic temperature and substrate specificity are discussed, and the starch utilization pathway in a broad range of temperatures is also proposed. PMID:25688178

  16. Fungal diversity in grape must and wine fermentation assessed by massive sequencing, quantitative PCR and DGGE.

    PubMed

    Wang, Chunxiao; García-Fernández, David; Mas, Albert; Esteve-Zarzoso, Braulio

    2015-01-01

    The diversity of fungi in grape must and during wine fermentation was investigated in this study by culture-dependent and culture-independent techniques. Carignan and Grenache grapes were harvested from three vineyards in the Priorat region (Spain) in 2012, and nine samples were selected from the grape must after crushing and during wine fermentation. From culture-dependent techniques, 362 isolates were randomly selected and identified by 5.8S-ITS-RFLP and 26S-D1/D2 sequencing. Meanwhile, genomic DNA was extracted directly from the nine samples and analyzed by qPCR, DGGE and massive sequencing. The results indicated that grape must after crushing harbored a high species richness of fungi with Aspergillus tubingensis, Aureobasidium pullulans, or Starmerella bacillaris as the dominant species. As fermentation proceeded, the species richness decreased, and yeasts such as Hanseniaspora uvarum, Starmerella bacillaris and Saccharomyces cerevisiae successively occupied the must samples. The "terroir" characteristics of the fungus population are more related to the location of the vineyard than to grape variety. Sulfur dioxide treatment caused a low effect on yeast diversity by similarity analysis. Because of the existence of large population of fungi on grape berries, massive sequencing was more appropriate to understand the fungal community in grape must after crushing than the other techniques used in this study. Suitable target sequences and databases were necessary for accurate evaluation of the community and the identification of species by the 454 pyrosequencing of amplicons. PMID:26557110

  17. Cellulose acetate based 3-dimensional electrospun scaffolds for skin tissue engineering applications.

    PubMed

    Atila, Deniz; Keskin, Dilek; Tezcaner, Ayşen

    2015-11-20

    Skin defects that are not able to regenerate by themselves are among the major problems faced. Tissue engineering approach holds promise for treating such defects. Development of tissue-mimicking-scaffolds that can promote healing process receives an increasing interest in recent years. In this study, 3-dimensional electrospun cellulose acetate (CA) pullulan (PULL) scaffolds were developed for the first time. PULL was intentionally used to obtain 3D structures with adjustable height. It was removed from the electrospun mesh to increase the porosity and biostability. Different ratios of the polymers were electrospun and analyzed with respect to degradation, porosity, and mechanical properties. It has been observed that fiber diameter, thickness and porosity of scaffolds increased with increased PULL content, on the other hand this resulted with higher degradation of scaffolds. Mechanical strength of scaffolds was improved after PULL removal suggesting their suitability as cell carriers. Cell culture studies were performed with the selected scaffold group (CA/PULL: 50/50) using mouse fibroblastic cell line (L929). In vitro cell culture tests showed that cells adhered, proliferated and populated CA/PULL (50/50) scaffolds showing that they are cytocompatible. Results suggest that uncrosslinked CA/PULL (50/50) electrospun scaffolds hold potential for skin tissue engineering applications.

  18. Characteristics of neutralization of acids by newly isolated fungal cells.

    PubMed

    Shiomi, Naofumi; Yasuda, Takako; Inoue, Yoko; Kusumoto, Noriko; Iwasaki, Saori; Katsuda, Tomohisa; Katoh, Shigeo

    2004-01-01

    Soil microorganisms play an important role in maintaining soil pH at levels suitable for other soil organisms. To clarify the biological neutralization mechanism in soil, we isolated soil microorganisms showing a high ability to neutralize acids and studied their characteristics. From our taxonomic study, three isolated strains were identified as filamentous fungi, namely Mucor sp., Aspergillus fumigatus, and Aureobasidium pullulans. These strains could secrete basic materials, such as ammonia, for neutralization, grow in the medium at pH 4.0 and increase the pH of the medium to approximately 8.0. These microbial cells could neutralize not only nitric acid but also sulfuric and hydrochloric acids. The strains could also grow by utilizing nitric acid as a sole nitrogen source. In the soil containing these organisms, the pH was maintained in the neutral range by the buffering action of basic materials that they secrete. These results suggest that these fungal cells are useful for protecting the soil from acidification by acid rain.

  19. Status, Antimicrobial Mechanism, and Regulation of Natural Preservatives in Livestock Food Systems

    PubMed Central

    Lee, Na-Kyoung; Paik, Hyun-Dong

    2016-01-01

    This review discusses the status, antimicrobial mechanisms, application, and regulation of natural preservatives in livestock food systems. Conventional preservatives are synthetic chemical substances including nitrates/nitrites, sulfites, sodium benzoate, propyl gallate, and potassium sorbate. The use of artificial preservatives is being reconsidered because of concerns relating to headache, allergies, and cancer. As the demand for biopreservation in food systems has increased, new natural antimicrobial compounds of various origins are being developed, including plant-derived products (polyphenolics, essential oils, plant antimicrobial peptides (pAMPs)), animal-derived products (lysozymes, lactoperoxidase, lactoferrin, ovotransferrin, antimicrobial peptide (AMP), chitosan and others), and microbial metabolites (nisin, natamycin, pullulan, ε-polylysine, organic acid, and others). These natural preservatives act by inhibiting microbial cell walls/membranes, DNA/RNA replication and transcription, protein synthesis, and metabolism. Natural preservatives have been recognized for their safety; however, these substances can influence color, smell, and toxicity in large amounts while being effective as a food preservative. Therefore, to evaluate the safety and toxicity of natural preservatives, various trials including combinations of other substances or different food preservation systems, and capsulation have been performed. Natamycin and nisin are currently the only natural preservatives being regulated, and other natural preservatives will have to be legally regulated before their widespread use.

  20. Status, Antimicrobial Mechanism, and Regulation of Natural Preservatives in Livestock Food Systems

    PubMed Central

    Lee, Na-Kyoung; Paik, Hyun-Dong

    2016-01-01

    This review discusses the status, antimicrobial mechanisms, application, and regulation of natural preservatives in livestock food systems. Conventional preservatives are synthetic chemical substances including nitrates/nitrites, sulfites, sodium benzoate, propyl gallate, and potassium sorbate. The use of artificial preservatives is being reconsidered because of concerns relating to headache, allergies, and cancer. As the demand for biopreservation in food systems has increased, new natural antimicrobial compounds of various origins are being developed, including plant-derived products (polyphenolics, essential oils, plant antimicrobial peptides (pAMPs)), animal-derived products (lysozymes, lactoperoxidase, lactoferrin, ovotransferrin, antimicrobial peptide (AMP), chitosan and others), and microbial metabolites (nisin, natamycin, pullulan, ε-polylysine, organic acid, and others). These natural preservatives act by inhibiting microbial cell walls/membranes, DNA/RNA replication and transcription, protein synthesis, and metabolism. Natural preservatives have been recognized for their safety; however, these substances can influence color, smell, and toxicity in large amounts while being effective as a food preservative. Therefore, to evaluate the safety and toxicity of natural preservatives, various trials including combinations of other substances or different food preservation systems, and capsulation have been performed. Natamycin and nisin are currently the only natural preservatives being regulated, and other natural preservatives will have to be legally regulated before their widespread use. PMID:27621697

  1. Combining mutualistic yeast and pathogenic virus--a novel method for codling moth control.

    PubMed

    Knight, Alan L; Witzgall, Peter

    2013-07-01

    The combination of a pathogenic virus and mutualistic yeasts isolated from larvae of codling moth Cydia pomonella is proposed as a novel insect control technique. Apples were treated with codling moth granulovirus (CpGV) and either one of three yeasts, Metschnikowia pulcherrima, Cryptococcus tephrensis, or Aureobasidium pullulans. The combination of yeasts with CpGV significantly increased mortality of neonate codling moth larvae, compared with CpGV alone. The three yeasts were equally efficient in enhancing the activity of CpGV. The addition of brown cane sugar to yeast further increased larval mortality and the protection of fruit against larvae. In comparison, without yeast, the addition of sugar to CpGV did not produce a significant effect. A field trial confirmed that fruit injury and larval survival were significantly reduced when apple trees were sprayed with CpGV, M. pulcherrima, and sugar. We have shown earlier that mutualistic yeasts are an essential part of codling moth larval diet. The finding that yeast also enhances larval ingestion of an insect-pathogenic virus is an opportunity for the development of a novel plant protection technique. We expect the combination of yeasts and insect pathogens to essentially contribute to future insect management.

  2. Analysis of Fungal Flora in Indoor Dust by Ribosomal DNA Sequence Analysis, Quantitative PCR, and Culture▿ †

    PubMed Central

    Pitkäranta, M.; Meklin, T.; Hyvärinen, A.; Paulin, L.; Auvinen, P.; Nevalainen, A.; Rintala, H.

    2008-01-01

    In recent years increasing attention has been given to the potential health effects of fungal exposure in indoor environments. We used large-scale sequencing of the fungal internal transcribed spacer region (ITS) of nuclear ribosomal DNA to describe the mycoflora of two office buildings over the four seasons. DNA sequencing was complemented by cultivation, ergosterol determination, and quantitative PCR analyses. Sequences of 1,339 clones were clustered into 394 nonredundant fungal operational taxonomical units containing sequences from 18 fungal subclasses. The observed flora differed markedly from that recovered by cultivation, the major differences being the near absence of several typical indoor mold genera such as Penicillium and Aspergillus spp. and a high prevalence of basidiomycetes in clone libraries. A total of 55% of the total diversity constituted of unidentifiable ITS sequences, some of which may represent novel fungal species. Dominant species were Cladosporium cladosporioides and C. herbarum, Cryptococcus victoriae, Leptosphaerulina americana and L. chartarum, Aureobasidium pullulans, Thekopsora areolata, Phaeococcomyces nigricans, Macrophoma sp., and several Malassezia species. Seasonal differences were observed for community composition, with ascomycetous molds and basidiomycetous yeasts predominating in the winter and spring and Agaricomycetidae basidiomycetes predominating in the fall. The comparison of methods suggested that the cloning, cultivation, and quantitative PCR methods complemented each other, generating a more comprehensive picture of fungal flora than any of the methods would give alone. The current restrictions of the methods are discussed. PMID:17981947

  3. The crosslinking of polysaccharides with polyamines and dextran-polyallylamine antibacterial hydrogels.

    PubMed

    O'Connor, Naphtali A; Abugharbieh, Ahmad; Yasmeen, Farzana; Buabeng, Emmanuel; Mathew, Steve; Samaroo, Diana; Cheng, Hai-Ping

    2015-01-01

    A facile modular approach to rapidly prepare pH-responsive hydrogels by crosslinking polysaccharides with polyamines is demonstrated. Hydrogels are prepared by first reacting the less reactive polysaccharides with the cross-linker epichlorohydrin and completed by the addition of polyamines. The crosslinking of polysaccharides with polyamines provides a facile method for incorporating functionality into polysaccharide based hydrogels. This process is demonstrated with the polysaccharides dextran, pullulan and carboxymethyl cellulose and with the polyamines polyallylamine and polyethylene imine. The hydrogels were characterized by FTIR and swelling studies, which showed pH-dependent swelling due to the presence of the polyamine. The hydrogels can also be tailored by varying the mass ratio between the polysaccharide and polyamine. Absorption studies of organic analytes showed the polyamine content affecting the uptake of a charged substrate (methylene blue) and no effect on a neutral substrate (6-methyl coumarin). This synthetic method was also used to prepare hydrogels with antibacterial activity against E. coli and S. aureus by utilizing an amphiphilic polyallylamine.

  4. Study on the effects of near-future ocean acidification on marine yeasts: a microcosm approach

    NASA Astrophysics Data System (ADS)

    Krause, Evamaria; Wichels, Antje; Erler, René; Gerdts, Gunnar

    2013-12-01

    Marine yeasts play an important role in biodegradation and nutrient cycling and are often associated with marine flora and fauna. They show maximum growth at pH levels lower than present-day seawater pH. Thus, contrary to many other marine organisms, they may actually profit from ocean acidification. Hence, we conducted a microcosm study, incubating natural seawater from the North Sea at present-day pH (8.10) and two near-future pH levels (7.81 and 7.67). Yeasts were isolated from the initial seawater sample and after 2 and 4 weeks of incubation. Isolates were classified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and representative isolates were identified by partial sequencing of the large subunit rRNA gene. From the initial seawater sample, we predominantly isolated a yeast-like filamentous fungus related to Aureobasidium pullulans, Cryptococcus sp., Candida sake, and various cold-adapted yeasts. After incubation, we found more different yeast species at near-future pH levels than at present-day pH. Yeasts reacting to low pH were related to Leucosporidium scottii, Rhodotorula mucilaginosa, Cryptococcus sp., and Debaryomyces hansenii. Our results suggest that these yeasts will benefit from seawater pH reductions and give a first indication that the importance of yeasts will increase in a more acidic ocean.

  5. Psychrophilic yeasts in glacial environments of Alpine glaciers.

    PubMed

    Turchetti, Benedetta; Buzzini, Pietro; Goretti, Marta; Branda, Eva; Diolaiuti, Guglielmina; D'Agata, Carlo; Smiraglia, Claudio; Vaughan-Martini, Ann

    2008-01-01

    The presence of psychrophilic yeasts in supra- and subglacial sediments, ice and meltwater collected from two glaciers of the Italian Alps (Forni and Sforzellina-Ortles-Cevedale group) was investigated. After incubation at 4 degrees C, subglacial sediments contained from 1.3 x 10(3) to 9.6 x 10(3) CFU of yeasts g(-1). The number of yeast cells in supraglacial sediments was c. 10-100-fold lower. A significant proportion of isolated yeasts exhibited one or more extracellular enzymatic activities (starch-degrading, lipolytic, esterolytic, proteolytic and pectinolytic activity) at 4 degrees C. Selected isolates were able to grow at 2 degrees C under laboratory-simulated in situ conditions. In all, 106 isolated yeasts were identified by MSP-PCR fingerprinting and 26S rRNA gene sequencing of the D1/D2 region as belonging to 10 species: Aureobasidium pullulans, Cryptococcus gilvescens (over 50% of the total), Cryptococcus terricolus, Mrakia gelida, Naganishia globosa, Rhodotorula glacialis, Rhodotorula psychrophenolica, Rhodotorula bacarum, Rhodotorula creatinivora and Rhodotorula laryngis. Four strains, all belonging to a new yeast species, yet to be described, were also isolated.

  6. Extracellular enzymatic activities and physiological profiles of yeasts colonizing fruit trees.

    PubMed

    Molnárová, Jana; Vadkertiová, Renáta; Stratilová, Eva

    2014-07-01

    Yeasts form a significant and diverse part of the phyllosphere microbiota. Some yeasts that inhabit plants have been found to exhibit extracellular enzymatic activities. The aim of the present study was to investigate the ability of yeasts isolated from leaves, fruits, and blossoms of fruit trees cultivated in Southwest Slovakia to produce extracellular enzymes, and to discover whether the yeasts originating from these plant organs differ from each other in their physiological properties. In total, 92 strains belonging to 29 different species were tested for: extracellular protease, β-glucosidase, lipase, and polygalacturonase activities; fermentation abilities; the assimilation of xylose, saccharose and alcohols (methanol, ethanol, glycerol); and for growth in a medium with 33% glucose. The black yeast Aureobasidium pullulans showed the largest spectrum of activities of all the species tested. Almost 70% of the strains tested demonstrated some enzymatic activity, and more than 90% utilized one of the carbon compounds tested. Intraspecies variations were found for the species of the genera Cryptococcus and Pseudozyma. Interspecies differences of strains exhibiting some enzymatic activities and utilizing alcohols were also noted. The largest proportion of the yeasts exhibited β-glucosidase activity and assimilated alcohols independently of their origin. The highest number of strains positive for all activities tested was found among the yeasts associated with leaves. Yeasts isolated from blossoms assimilated saccharose and D-xylose the most frequently of all the yeasts tested. The majority of the fruit-inhabiting yeasts grew in the medium with higher osmotic pressure.

  7. Nanogel-based PspA intranasal vaccine prevents invasive disease and nasal colonization by Streptococcus pneumoniae.

    PubMed

    Kong, Il Gyu; Sato, Ayuko; Yuki, Yoshikazu; Nochi, Tomonori; Takahashi, Haruko; Sawada, Shinichi; Mejima, Mio; Kurokawa, Shiho; Okada, Kazunari; Sato, Shintaro; Briles, David E; Kunisawa, Jun; Inoue, Yusuke; Yamamoto, Masafumi; Akiyoshi, Kazunari; Kiyono, Hiroshi

    2013-05-01

    To establish a safer and more effective vaccine against pneumococcal respiratory infections, current knowledge regarding the antigens common among pneumococcal strains and improvements to the system for delivering these antigens across the mucosal barrier must be integrated. We developed a pneumococcal vaccine that combines the advantages of pneumococcal surface protein A (PspA) with a nontoxic intranasal vaccine delivery system based on a nanometer-sized hydrogel (nanogel) consisting of a cationic cholesteryl group-bearing pullulan (cCHP). The efficacy of the nanogel-based PspA nasal vaccine (cCHP-PspA) was tested in murine pneumococcal airway infection models. Intranasal vaccination with cCHP-PspA provided protective immunity against lethal challenge with Streptococcus pneumoniae Xen10, reduced colonization and invasion by bacteria in the upper and lower respiratory tracts, and induced systemic and nasal mucosal Th17 responses, high levels of PspA-specific serum immunoglobulin G (IgG), and nasal and bronchial IgA antibody responses. Moreover, there was no sign of PspA delivery by nanogel to either the olfactory bulbs or the central nervous system after intranasal administration. These results demonstrate the effectiveness and safety of the nanogel-based PspA nasal vaccine system as a universal mucosal vaccine against pneumococcal respiratory infection. PMID:23460513

  8. Nanogel-based pneumococcal surface protein A nasal vaccine induces microRNA-associated Th17 cell responses with neutralizing antibodies against Streptococcus pneumoniae in macaques.

    PubMed

    Fukuyama, Y; Yuki, Y; Katakai, Y; Harada, N; Takahashi, H; Takeda, S; Mejima, M; Joo, S; Kurokawa, S; Sawada, S; Shibata, H; Park, E J; Fujihashi, K; Briles, D E; Yasutomi, Y; Tsukada, H; Akiyoshi, K; Kiyono, H

    2015-09-01

    We previously established a nanosized nasal vaccine delivery system by using a cationic cholesteryl group-bearing pullulan nanogel (cCHP nanogel), which is a universal protein-based antigen-delivery vehicle for adjuvant-free nasal vaccination. In the present study, we examined the central nervous system safety and efficacy of nasal vaccination with our developed cCHP nanogel containing pneumococcal surface protein A (PspA-nanogel) against pneumococcal infection in nonhuman primates. When [(18)F]-labeled PspA-nanogel was nasally administered to a rhesus macaque (Macaca mulatta), longer-term retention of PspA was noted in the nasal cavity when compared with administration of PspA alone. Of importance, no deposition of [(18)F]-PspA was seen in the olfactory bulbs or brain. Nasal PspA-nanogel vaccination effectively induced PspA-specific serum IgG with protective activity and mucosal secretory IgA (SIgA) Ab responses in cynomolgus macaques (Macaca fascicularis). Nasal PspA-nanogel-induced immune responses were mediated through T-helper (Th) 2 and Th17 cytokine responses concomitantly with marked increases in the levels of miR-181a and miR-326 in the serum and respiratory tract tissues, respectively, of the macaques. These results demonstrate that nasal PspA-nanogel vaccination is a safe and effective strategy for the development of a nasal vaccine for the prevention of pneumonia in humans. PMID:25669148

  9. Nanogel-based pneumococcal surface protein A nasal vaccine induces microRNA-associated Th17 cell responses with neutralizing antibodies against Streptococcus pneumoniae in macaques

    PubMed Central

    Fukuyama, Y; Yuki, Y; Katakai, Y; Harada, N; Takahashi, H; Takeda, S; Mejima, M; Joo, S; Kurokawa, S; Sawada, S; Shibata, H; Park, E J; Fujihashi, K; Briles, D E; Yasutomi, Y; Tsukada, H; Akiyoshi, K; Kiyono, H

    2015-01-01

    We previously established a nanosized nasal vaccine delivery system by using a cationic cholesteryl group-bearing pullulan nanogel (cCHP nanogel), which is a universal protein-based antigen-delivery vehicle for adjuvant-free nasal vaccination. In the present study, we examined the central nervous system safety and efficacy of nasal vaccination with our developed cCHP nanogel containing pneumococcal surface protein A (PspA-nanogel) against pneumococcal infection in nonhuman primates. When [18F]-labeled PspA-nanogel was nasally administered to a rhesus macaque (Macaca mulatta), longer-term retention of PspA was noted in the nasal cavity when compared with administration of PspA alone. Of importance, no deposition of [18F]-PspA was seen in the olfactory bulbs or brain. Nasal PspA-nanogel vaccination effectively induced PspA-specific serum IgG with protective activity and mucosal secretory IgA (SIgA) Ab responses in cynomolgus macaques (Macaca fascicularis). Nasal PspA-nanogel-induced immune responses were mediated through T-helper (Th) 2 and Th17 cytokine responses concomitantly with marked increases in the levels of miR-181a and miR-326 in the serum and respiratory tract tissues, respectively, of the macaques. These results demonstrate that nasal PspA-nanogel vaccination is a safe and effective strategy for the development of a nasal vaccine for the prevention of pneumonia in humans. PMID:25669148

  10. Nanogel-Based PspA Intranasal Vaccine Prevents Invasive Disease and Nasal Colonization by Streptococcus pneumoniae

    PubMed Central

    Kong, Il Gyu; Sato, Ayuko; Nochi, Tomonori; Takahashi, Haruko; Sawada, Shinichi; Mejima, Mio; Kurokawa, Shiho; Okada, Kazunari; Sato, Shintaro; Briles, David E.; Kunisawa, Jun; Inoue, Yusuke; Yamamoto, Masafumi; Akiyoshi, Kazunari

    2013-01-01

    To establish a safer and more effective vaccine against pneumococcal respiratory infections, current knowledge regarding the antigens common among pneumococcal strains and improvements to the system for delivering these antigens across the mucosal barrier must be integrated. We developed a pneumococcal vaccine that combines the advantages of pneumococcal surface protein A (PspA) with a nontoxic intranasal vaccine delivery system based on a nanometer-sized hydrogel (nanogel) consisting of a cationic cholesteryl group-bearing pullulan (cCHP). The efficacy of the nanogel-based PspA nasal vaccine (cCHP-PspA) was tested in murine pneumococcal airway infection models. Intranasal vaccination with cCHP-PspA provided protective immunity against lethal challenge with Streptococcus pneumoniae Xen10, reduced colonization and invasion by bacteria in the upper and lower respiratory tracts, and induced systemic and nasal mucosal Th17 responses, high levels of PspA-specific serum immunoglobulin G (IgG), and nasal and bronchial IgA antibody responses. Moreover, there was no sign of PspA delivery by nanogel to either the olfactory bulbs or the central nervous system after intranasal administration. These results demonstrate the effectiveness and safety of the nanogel-based PspA nasal vaccine system as a universal mucosal vaccine against pneumococcal respiratory infection. PMID:23460513

  11. Nanogel antigenic protein-delivery system for adjuvant-free intranasal vaccines.

    PubMed

    Nochi, Tomonori; Yuki, Yoshikazu; Takahashi, Haruko; Sawada, Shin-ichi; Mejima, Mio; Kohda, Tomoko; Harada, Norihiro; Kong, Il Gyu; Sato, Ayuko; Kataoka, Nobuhiro; Tokuhara, Daisuke; Kurokawa, Shiho; Takahashi, Yuko; Tsukada, Hideo; Kozaki, Shunji; Akiyoshi, Kazunari; Kiyono, Hiroshi

    2010-07-01

    Nanotechnology is an innovative method of freely controlling nanometre-sized materials. Recent outbreaks of mucosal infectious diseases have increased the demands for development of mucosal vaccines because they induce both systemic and mucosal antigen-specific immune responses. Here we developed an intranasal vaccine-delivery system with a nanometre-sized hydrogel ('nanogel') consisting of a cationic type of cholesteryl-group-bearing pullulan (cCHP). A non-toxic subunit fragment of Clostridium botulinum type-A neurotoxin BoHc/A administered intranasally with cCHP nanogel (cCHP-BoHc/A) continuously adhered to the nasal epithelium and was effectively taken up by mucosal dendritic cells after its release from the cCHP nanogel. Vigorous botulinum-neurotoxin-A-neutralizing serum IgG and secretory IgA antibody responses were induced without co-administration of mucosal adjuvant. Importantly, intranasally administered cCHP-BoHc/A did not accumulate in the olfactory bulbs or brain. Moreover, intranasally immunized tetanus toxoid with cCHP nanogel induced strong tetanus-toxoid-specific systemic and mucosal immune responses. These results indicate that cCHP nanogel can be used as a universal protein-based antigen-delivery vehicle for adjuvant-free intranasal vaccination. PMID:20562880

  12. Carbohydrate binding properties of banana (Musa acuminata) lectin I. Novel recognition of internal alpha1,3-linked glucosyl residues.

    PubMed

    Mo, H; Winter, H C; Van Damme, E J; Peumans, W J; Misaki, A; Goldstein, I J

    2001-05-01

    Examination of lectins of banana (Musa acuminata) and the closely related plantain (Musa spp.) by the techniques of quantitative precipitation, hapten inhibition of precipitation, and isothermal titration calorimetry showed that they are mannose/glucose binding proteins with a preference for the alpha-anomeric form of these sugars. Both generate precipitin curves with branched chain alpha-mannans (yeast mannans) and alpha-glucans (glycogens, dextrans, and starches), but not with linear alpha-glucans containing only alpha1,4- and alpha1,6-glucosidic bonds (isolichenan and pullulan). The novel observation was made that banana and plantain lectins recognize internal alpha1,3-linked glucosyl residues, which occur in the linear polysaccharides elsinan and nigeran. Concanavalin A and lectins from pea and lentil, also mannose/glucose binding lectins, did not precipitate with any of these linear alpha-glucans. This is, the authors believe, the first report of the recognition of internal alpha1,3-glucosidic bonds by a plant lectin. It is possible that these lectins are present in the pulp of their respective fruit, complexed with starch.

  13. [Diversity and community structure of endophytic fungi from Taxus chinensis var. mairei].

    PubMed

    2014-07-01

    A total of 628 endophytic fungi were isolated from 480 tissue segments of needles and branches of Taxus chinensis var. mairei. According to morphological characteristics and ITS sequences, they represented 43 taxa in 28 genera, of which 10 Hyphomycetes, 20 Coelomycetes, 12 Ascomycetes and 1 unknown fungus. Phomopsis mali was confirmed as the dominant species. In accordance with relative frequency, Alternaria alternata, Aureobasidium pullulans, Colletotrichum boninense, C. gloeosporioides, Epicoccum nigrum , Fungal sp., Fusarium lateritium, Glomerella cingulata, Magnaporthales sp. , Nigrospora oryzae, Pestalotiopsis maculiformans, P. microspora, Peyronellaea glomerata and Xylaria sp. 1 were more common in T. chinensis var. mairei. T. chinensis var. mairei were severely infected by endophytic fungi. Endophytic fungi were found in 81 percent of plant tissues with a high diversity. Distribution ranges of endophytic fungi were influenced by tissue properties. The colonization rate, richness, diversity of endophytic fungi in needles were obviously lower than in branches, and kinds of endophytic fungi between branches were more similar than those in needles, thus endophytic fungi had tissue preference. In addition, tissue age influenced the community structure of endophytic fungi. The elder branch tissues were, the higher colonization rate, richness, diversity of endophytic fungi were. Systematic studying the diversity and community structure of endophytic fungi in T. chinensis var. mairei and clarifying their distribution regularity in plant tissues would offer basic data and scientific basis for their development and utilization. Discussing the presence of fungal pathogens in healthy plant tissues would be of positive significance for source protection of T. chinensis var. mairei.

  14. Fungal diversity in grape must and wine fermentation assessed by massive sequencing, quantitative PCR and DGGE

    PubMed Central

    Wang, Chunxiao; García-Fernández, David; Mas, Albert; Esteve-Zarzoso, Braulio

    2015-01-01

    The diversity of fungi in grape must and during wine fermentation was investigated in this study by culture-dependent and culture-independent techniques. Carignan and Grenache grapes were harvested from three vineyards in the Priorat region (Spain) in 2012, and nine samples were selected from the grape must after crushing and during wine fermentation. From culture-dependent techniques, 362 isolates were randomly selected and identified by 5.8S-ITS-RFLP and 26S-D1/D2 sequencing. Meanwhile, genomic DNA was extracted directly from the nine samples and analyzed by qPCR, DGGE and massive sequencing. The results indicated that grape must after crushing harbored a high species richness of fungi with Aspergillus tubingensis, Aureobasidium pullulans, or Starmerella bacillaris as the dominant species. As fermentation proceeded, the species richness decreased, and yeasts such as Hanseniaspora uvarum, Starmerella bacillaris and Saccharomyces cerevisiae successively occupied the must samples. The “terroir” characteristics of the fungus population are more related to the location of the vineyard than to grape variety. Sulfur dioxide treatment caused a low effect on yeast diversity by similarity analysis. Because of the existence of large population of fungi on grape berries, massive sequencing was more appropriate to understand the fungal community in grape must after crushing than the other techniques used in this study. Suitable target sequences and databases were necessary for accurate evaluation of the community and the identification of species by the 454 pyrosequencing of amplicons. PMID:26557110

  15. Humidifier lung and humidifier fever.

    PubMed

    Baur, X; Behr, J; Dewair, M; Ehret, W; Fruhmann, G; Vogelmeier, C; Weiss, W; Zinkernagel, V

    1988-01-01

    The aim of our study was to evaluate clinical diagnoses in symptomatic persons exposed to aerosols from humidifiers or air conditioners. In addition, we tried to identify the causative antigens. Results of clinical investigations, including inhalation challenge tests, demonstrated a typical hypersensitivity pneumonitis (humidifier lung) in 9 persons and isolated systemic symptoms without significant changes in lung function and chest x-rays (humidifier fever) in 3 persons. Microbiological studies revealed a variety of fungi and bacteria in the water supplies of humidifiers and air conditioners at patients' workplaces. The detection of 4 members of the order Sphaeropsidales (Deuteromycotina), not previously associated with humidifier-induced diseases, is of special interest. By means of an improved polystyrene tube-immunoradiometric assay, high concentrations of IgG antibodies against extracts prepared from water of patients' humidifier systems were found in all cases. In addition, patients demonstrated low concentrations of IgG antibodies against thermophilic Actinomycetes, and usually also against various fungi, such as Alternaria tenuis, Aureobasidium pullulans, Penicillium notatum, Aspergilli, and fungi of the order Sphaeropsidales, which were isolated and cultured from humidifier water supplies. The much higher concentrations of antibodies against humidifier/air conditioner water extracts seem to result from sensitization to a variety of antigens from different fungi and bacteria. For in vivo and in vitro diagnostic tests in humidifier-induced lung diseases, we especially recommend using extracts from water systems installed at the corresponding workplaces. PMID:3130530

  16. Humidifier lung and humidifier fever.

    PubMed

    Baur, X; Behr, J; Dewair, M; Ehret, W; Fruhmann, G; Vogelmeier, C; Weiss, W; Zinkernagel, V

    1988-12-01

    The aim of our study was to evaluate clinical diagnoses in symptomatic persons exposed to aerosols from humidifiers or air conditioners. In addition, we tried to identify the causative antigens. Results of clinical investigations, including inhalation challenge tests, demonstrated a typical hypersensitivity pneumonitis (humidifier lung) in 9 persons and isolated systemic symptoms without significant changes in lung function and chest x-rays (humidifier fever) in 3 persons. Microbiological studies revealed a variety of fungi and bacteria in the water supplies of humidifiers and air conditioners at patients' workplaces. The detection of 4 members of the orderSphaeropsidales (Deuteromycotina), not previously associated with humidifier-induced diseases, is of special interest. By means of an improved polystyrene tube-immunoradiometric assay, high concentrations of IgG antibodies against extracts prepared from water of patients' humidifier systems were found in all cases. In addition, patients demonstrated low concentrations of IgG antibodies against thermophilicActinomycetes, and usually also against various fungi, such asAlternaria tenuis, Aureobasidium pullulans, Penicillium notatum, Aspergilli, and fungi of the orderSphaeropsidales, which were isolated and cultured from humidifier water supplies. The much higher concentrations of antibodies against humidifier/air conditioner water extracts seem to result from sensitization to a variety of antigens from different fungi and bacteria. For in vivo and in vitro diagnostic tests in humidifier-induced lung diseases, we especially recommend using extracts from water systems installed at the corresponding workplaces. PMID:27519372

  17. Increased prevalence of IgG-induced sensitization and hypersensitivity pneumonitis (humidifier lung) in nonsmokers exposed to aerosols of a contaminated air conditioner.

    PubMed

    Baur, X; Richter, G; Pethran, A; Czuppon, A B; Schwaiblmair, M

    1992-01-01

    Specific IgG antibodies against antigens of a contaminated air conditioner were estimated in serum of 134 workers of a printing company. Altogether 64% of the workers investigated revealed significantly elevated levels (> 3 U/ml) of IgG antibodies specific to these antigens as compared to a nonexposed control group. The occurrence of IgG antibodies for microbial extracts were 25% for Fusarium, 23% for Penicillium notatum, 13% for Alternaria tenuis, 12% for Aureobasidium pullulans, 9% for Sphaeropsidales species, 3% for Micropolyspora faeni, 2% for Aspergillus fumigatus and 2% for Thermoactionomyces vulgaris. Out of the 86 workers with elevated IgG antibodies for air conditioner antigens, 59 were nonsmokers. Considering a cut-off level of 10 U/ml IgG for high values, the proportion of smokers to nonsmokers becomes even more pronounced (6 to 36 respectively, binominal test p < 0.001). This is despite the fact that the distribution of smokers and nonsmokers among the 134 workers is approximately equal (60 to 74). All 3 workers with clinical diagnosis of humidifier lung or humidifier fever belonged to the nonsmoker group. Our findings indicate that crude water extracts of contaminated air conditioners are the best choice as antigen source for the diagnosis of humidifier lung in exposed workers. Nonsmokers are shown to have a high risk for immunological sensitization. PMID:1485005

  18. Preparation of linear maltodextrins using a hyperthermophilic amylopullulanase with cyclodextrin- and starch-hydrolysing activities.

    PubMed

    Li, Xiaolei; Li, Dan

    2015-03-30

    A novel method for the preparation of linear maltodextrins from cyclodextrins and starch was proposed. To accomplish this process, an amylopullulanase from hyperthermophilic archaeon Caldivirga maquilingensis (CMApu) was characterized and used. CMApu with an estimated molecular mass of 62.7 kDa by SDS-PAGE had a maximal pullulan-hydrolysing activity at 100°C and pH 5.0. It could also hydrolyse amylopectin (AP), starch, β-CD and amylose (AM), in a decreasing order of relative activities from 88.96% to 57.17%. TLC and HPAEC analysis revealed that CMApu catalyzed the debranching and degrading reactions to produce linear malto-oligosaccharides (≤ G8-G1) from G8-β-CD and/or normal CDs, amylodextrins (DP6-96) from AM, and amylodextrins (DP1-76) from AP and potato starch. Our results showed that CMApu had a great potential for the industrial preparation of linear maltodextrins from normal starch instead of waxy starch, malto-oligosaccharides or sucrose. And the high optimal temperature of CMApu facilitated the simultaneous gelatinization and hydrolysis of cereal starch.

  19. Nanogel antigenic protein-delivery system for adjuvant-free intranasal vaccines

    NASA Astrophysics Data System (ADS)

    Nochi, Tomonori; Yuki, Yoshikazu; Takahashi, Haruko; Sawada, Shin-Ichi; Mejima, Mio; Kohda, Tomoko; Harada, Norihiro; Kong, Il Gyu; Sato, Ayuko; Kataoka, Nobuhiro; Tokuhara, Daisuke; Kurokawa, Shiho; Takahashi, Yuko; Tsukada, Hideo; Kozaki, Shunji; Akiyoshi, Kazunari; Kiyono, Hiroshi

    2010-07-01

    Nanotechnology is an innovative method of freely controlling nanometre-sized materials. Recent outbreaks of mucosal infectious diseases have increased the demands for development of mucosal vaccines because they induce both systemic and mucosal antigen-specific immune responses. Here we developed an intranasal vaccine-delivery system with a nanometre-sized hydrogel (`nanogel') consisting of a cationic type of cholesteryl-group-bearing pullulan (cCHP). A non-toxic subunit fragment of Clostridium botulinum type-A neurotoxin BoHc/A administered intranasally with cCHP nanogel (cCHP-BoHc/A) continuously adhered to the nasal epithelium and was effectively taken up by mucosal dendritic cells after its release from the cCHP nanogel. Vigorous botulinum-neurotoxin-A-neutralizing serum IgG and secretory IgA antibody responses were induced without co-administration of mucosal adjuvant. Importantly, intranasally administered cCHP-BoHc/A did not accumulate in the olfactory bulbs or brain. Moreover, intranasally immunized tetanus toxoid with cCHP nanogel induced strong tetanus-toxoid-specific systemic and mucosal immune responses. These results indicate that cCHP nanogel can be used as a universal protein-based antigen-delivery vehicle for adjuvant-free intranasal vaccination.

  20. Hydrocarbons, the advanced biofuels produced by different organisms, the evidence that alkanes in petroleum can be renewable.

    PubMed

    Fu, Wen-Juan; Chi, Zhe; Ma, Zai-Chao; Zhou, Hai-Xiang; Liu, Guang-Lei; Lee, Ching-Fu; Chi, Zhen-Ming

    2015-09-01

    It is generally regarded that the petroleum cannot be renewable. However, in recent years, it has been found that many marine cyanobacteria, some eubacteria, engineered Escherichia coli, some endophytic fungi, engineered yeasts, some marine yeasts, plants, and insects can synthesize hydrocarbons with different carbon lengths. If the organisms, especially some native microorganisms and engineered bacteria and yeasts, can synthesize and secret a large amount of hydrocarbons within a short period, alkanes in the petroleum can be renewable. It has been documented that there are eight pathways for hydrocarbon biosynthesis in different organisms. Unfortunately, most of native microorganisms, engineered E. coli and engineered yeasts, only synthesize a small amount of intracellular and extracellular hydrocarbons. Recently, Aureobasidium pullulans var. melanogenum isolated from a mangrove ecosystem has been found to be able to synthesize and secret over 21.5 g/l long-chain hydrocarbons with a yield of 0.275 g/g glucose and a productivity of 0.193 g/l/h within 5 days. The yeast may have highly potential applications in alkane production.

  1. Biodiversity and safety aspects of yeast strains characterized from vineyards and spontaneous fermentations in the Apulia Region, Italy.

    PubMed

    Tristezza, Mariana; Vetrano, Cosimo; Bleve, Gianluca; Spano, Giuseppe; Capozzi, Vittorio; Logrieco, Antonio; Mita, Giovanni; Grieco, Francesco

    2013-12-01

    This work is the first large-scale study on vineyard-associated yeast strains from Apulia (Southern Italy). Yeasts were identified by Internal Transcribed Spacer (ITS) ribotyping and bioinformatic analysis. The polymorphism of interdelta elements was used to differentiate Saccharomyces cerevisiae strains. Twenty different species belonging to 9 genera were identified. Predominant on the grape surface were Metschnikowia pulcherrima, Hanseniaspora uvarum and Aureobasidium pullulans, whereas M. pulcherrima and H. uvarum were dominant in the early fermentation stage. A total of 692 S. cerevisiae isolates were identified and a number of S. cerevisiae strains, ranging from 26 to 55, was detected in each of the eight fermentations. The strains were tested for biogenic amines (BAs) production, either in synthetic media or grape must. Two Pichia manshurica, an Issatchenkia terricola and a M. pulcherrima strains were able to produce histamine and cadaverine, during must fermentation. The production of BAs in wine must was different than that observed in the synthetic medium. This feature indicate the importance of an "in grape must" assessment of BAs producing yeast. Overall, our results suggest the importance of microbiological control during wine-making to reduce the potential health risk for consumer represented by these spoilage yeasts.

  2. Recombinant expression, activity screening and functional characterization identifies three novel endo-1,4-β-glucanases that efficiently hydrolyse cellulosic substrates.

    PubMed

    Tambor, José Humberto; Ren, Huanan; Ushinsky, Sophia; Zheng, Yun; Riemens, Anja; St-Francois, Christopher; Tsang, Adrian; Powlowski, Justin; Storms, Reginald

    2012-01-01

    The hydrolysis of cellulose into fermentable sugars is a costly and rate-limiting step in the production of biofuels from renewable feedstocks. Developing new cellulase systems capable of increased cellulose hydrolysis rates would reduce biofuel production costs. With this in mind, we screened 55 fungal endoglucanases for their abilities to be expressed at high levels by Aspergillus niger and to hydrolyze amorphous cellulose at rates significantly greater than that obtained with TrCel5A, one of the major endoglucanases in the Trichoderma reesei cellulase system. This screen identified three endoglucanases, Aureobasidium pullulans ApCel5A, Gloeophyllum trabeum GtCel12A and Sporotrichum thermophile StCel5A. We determined that A. niger expressed the three endoglucanases at relatively high levels (≥0.3 g/l) and that the hydrolysis rate of ApCel5A and StCel5A with carboxymethylcellulose 4M as substrate was five and two times greater than the T. reesei Cel5A. The ApCel5A, GtCel12A and StCel5A enzymes also demonstrated significant synergy with Cel7A/CbhI, the major exoglucanase in the T. reesei cellulase system. The three endoglucanases characterized in this study are, therefore, promising candidate endoglucanases for developing new cellulase systems with increased rates of cellulose saccharification.

  3. Design, Synthesis, and Photophysical Properties of Pyrroloquinoline-Based Compounds Showing Strong Blue Fluorescence as Potential Dyes for Biomedical Applications.

    PubMed

    Carta, Davide; Balasso, Anna; Caliceti, Paolo; Ferlin, Maria Grazia

    2015-11-01

    A small library of 3-ethylpyrrolo[3,2-f]quinoline derivatives was synthesized to identify a novel class of dyes for use in biological studies. According to the spectroscopic analyses performed to evaluate the fluorimetric parameters of quantum yield and brightness, 7-methyl- and 6,7-dimethylpyrroloquinolin(9)one derivatives were found to be the best blue luminescent dyes for biological applications. To enhance the luminescence profiles and to obtain probes that could be conjugated to functional groups of supramolecular drug delivery systems, these compounds were further modified at position 3 to obtain 3-heptanoic acid and 3-aminohexylpyrroloquinolin(9)one methylated derivatives. The most brilliant 6,7-dimethyl-3-aminohexylpyrroloquinolinone hydrochloride was conjugated to pullulan, a biocompatible polysaccharide used to produce colloidal systems for drug delivery. Comparative studies showed that this compound can be properly exploited as a blue fluorescent label in biological investigations, namely cell trafficking and pharmacokinetics/biodistribution studies. These molecules possess higher fluorescence efficiency than commercial dyes in biological media, making them suitable alternatives to commercially available products in current use. PMID:26447862

  4. Structure and function of α-glucan debranching enzymes.

    PubMed

    Møller, Marie Sofie; Henriksen, Anette; Svensson, Birte

    2016-07-01

    α-Glucan debranching enzymes hydrolyse α-1,6-linkages in starch/glycogen, thereby, playing a central role in energy metabolism in all living organisms. They belong to glycoside hydrolase families GH13 and GH57 and several of these enzymes are industrially important. Nine GH13 subfamilies include α-glucan debranching enzymes; isoamylase and glycogen debranching enzymes (GH13_11); pullulanase type I/limit dextrinase (GH13_12-14); pullulan hydrolase (GH13_20); bifunctional glycogen debranching enzyme (GH13_25); oligo-1 and glucan-1,6-α-glucosidases (GH13_31); pullulanase type II (GH13_39); and α-amylase domains (GH13_41) in two-domain amylase-pullulanases. GH57 harbours type II pullulanases. Specificity differences, domain organisation, carbohydrate binding modules, sequence motifs, three-dimensional structures and specificity determinants are discussed. The phylogenetic analysis indicated that GH13_39 enzymes could represent a "missing link" between the strictly α-1,6-specific debranching enzymes and the enzymes with dual specificity and α-1,4-linkage preference.

  5. Biopolymer-Based Nanoparticles for Drug/Gene Delivery and Tissue Engineering

    PubMed Central

    Nitta, Sachiko Kaihara; Numata, Keiji

    2013-01-01

    There has been a great interest in application of nanoparticles as biomaterials for delivery of therapeutic molecules such as drugs and genes, and for tissue engineering. In particular, biopolymers are suitable materials as nanoparticles for clinical application due to their versatile traits, including biocompatibility, biodegradability and low immunogenicity. Biopolymers are polymers that are produced from living organisms, which are classified in three groups: polysaccharides, proteins and nucleic acids. It is important to control particle size, charge, morphology of surface and release rate of loaded molecules to use biopolymer-based nanoparticles as drug/gene delivery carriers. To obtain a nano-carrier for therapeutic purposes, a variety of materials and preparation process has been attempted. This review focuses on fabrication of biocompatible nanoparticles consisting of biopolymers such as protein (silk, collagen, gelatin, β-casein, zein and albumin), protein-mimicked polypeptides and polysaccharides (chitosan, alginate, pullulan, starch and heparin). The effects of the nature of the materials and the fabrication process on the characteristics of the nanoparticles are described. In addition, their application as delivery carriers of therapeutic drugs and genes and biomaterials for tissue engineering are also reviewed. PMID:23344060

  6. Three novel NY-ESO-1 epitopes bound to DRB1*0803, DQB1*0401 and DRB1*0901 recognized by CD4 T cells from CHP-NY-ESO-1-vaccinated patients.

    PubMed

    Mizote, Yu; Taniguchi, Taku; Tanaka, Kei; Isobe, Midori; Wada, Hisashi; Saika, Takashi; Kita, Shoichi; Koide, Yukari; Uenaka, Akiko; Nakayama, Eiichi

    2010-07-19

    Three novel NY-ESO-1 CD4 T cell epitopes were identified using PBMC obtained from patients who were vaccinated with a complex of cholesterol-bearing hydrophobized pullulan (CHP) and NY-ESO-1 protein (CHP-NY-ESO-1). The restriction molecules were determined by antibody blocking and using various EBV-B cells with different HLA alleles as APC to present peptides to CD4 T cells. The minimal epitope peptides were determined using various N- and C-termini truncated peptides deduced from 18-mer overlapping peptides originally identified for recognition. Those epitopes were DRB1*0901-restricted NY-ESO-1 87-100, DQB1*0401-restricted NY-ESO-1 95-107 and DRB1*0803-restricted NY-ESO-1 124-134. CD4 T cells used to determine those epitope peptides recognized EBV-B cells or DC that were treated with recombinant NY-ESO-1 protein or NY-ESO-1-expressing tumor cell lysate, suggesting that the epitope peptides are naturally processed. These CD4 T cells showed a cytokine profile with Th1 characteristics. Furthermore, NY-ESO-1 87-100 peptide/HLA-DRB1*0901 tetramer staining was observed. Multiple Th1-type CD4 T cell responses are beneficial for inducing effective anti-tumor responses after NY-ESO-1 protein vaccination.

  7. Halophilic alkali- and thermostable amylase from a novel polyextremophilic Amphibacillus sp. NM-Ra2.

    PubMed

    Mesbah, Noha M; Wiegel, Juergen

    2014-09-01

    Extracellular gluco-amylo-pullulanase from Amphibacillus sp. NM-Ra2 was purified to homogeneity by ethanol precipitation, anion exchange chromatography and gel filtration chromatography. Molecular mass of the enzyme was 50kDa (SDS-PAGE). The enzyme showed maximal activity at 1.9 M NaCl, pH50°C 8.0 and 54°C and was active from 0 to 4.3 M NaCl and 37 to 65°C. The enzyme was inhibited by EDTA and was stable and active in the presence of PMSF, DTT, H2O2, Triton-X-100, Tween 20 and Tween 80. Ca2+ is inessential for activity. The amylase was stimulated with K+ and inhibited with Cu2+ and Mg2+. Hg2+, Zn2+ and Fe2+ had no effect on activity. Amylase was stable and active in the presence of ethanol, methanol and benzene (25%, v/v). The enzyme hydrolyzed linear and branched polysaccharides including pullulan, glycogen and amylopectin, and hydrolyzed raw wheat starch and raw corn starch (14.6% and 13.5% over 2 h). Amylase activity was inhibited by soluble starch concentrations greater than 0.3%. The major products of soluble starch hydrolysis were maltose and maltotriose. The amylase, being halophilic and alkali-thermostable, in addition to being resistant to surfactants, oxidizing agents and organic solvents, can find applications in the starch processing, pharmaceutical, food and paper/pulp industries.

  8. Formulation and stability evaluation of 3D alginate beads potentially useful for cumulus-oocyte complexes culture.

    PubMed

    Dorati, Rossella; Genta, Ida; Ferrari, Michela; Vigone, Giulia; Merico, Valeria; Garagna, Silvia; Zuccotti, Maurizio; Conti, Bice

    2016-01-01

    Ovarian follicle encapsulation in synthetic or natural matrixes based on biopolymers is potentially a promising approach to in vitro maturation (IVM) process, since it maintains follicle 3D organisation by preventing its flattening and consequent disruption of gap junctions, preserving the functional relationship between oocyte and companion follicle cells. The aim of the work was to optimise physico-chemical parameters of alginate microcapsules for perspective IVM under 3D environments. On this purpose alginate and cross-linking agent concentrations were investigated. Alginate concentration between 0.75% and 0.125% w/w and Mg(2+), Ba(2+), Ca(2+ )at concentration between 100 and 20 mM were tested. Follicle encapsulation was obtained by on purpose modified diffusion setting gelation technique, and evaluated together with beads, chemical and mechanical stability in standard and stressing conditions. Beads permeability was tested towards albumin, fetuin, pyruvate, glucose, pullulan. Results demonstrated that 0.25% alginate cross-linked in 100 mM CaCl2 beads is suitable to follicle encapsulation. PMID:26791322

  9. Purification and characterisation of a malto-oligosaccharide-forming amylase active at high pH from Bacillus clausii BT-21.

    PubMed

    Duedahl-Olesen, L; Kragh, K M; Zimmermann, W

    2000-10-20

    Bacillus clausii BT-21 produced an extracellular malto-oligosaccharide-forming amylase active at high pH when grown on starch substrates. The enzyme was purified to homogeneity by affinity and anion-exchange chromatography. The molecular weight of the enzyme estimated by sodium dodecyl sulfate polyacrylamide electrophoresis was 101 kDa. The enzyme showed an optimum of activity at pH 9.5 and 55 degrees C. Maltohexaose was detected as the main initially formed starch hydrolysis product. Maltotetraose and maltose were the main products obtained after hydrolysis of starch by the enzyme for an extended period of time and were not further degraded. The enzyme readily hydrolysed soluble starch, amylopectin and amylose, while cyclodextrins, pullulan or dextran were not degraded. The mode of action during hydrolysis of starch indicated an exo-acting type of amylolytic enzyme mainly producing maltohexaose and maltotetraose. Amino acid sequencing of the enzyme revealed high homology with the maltohexaose-forming amylase from Bacillus sp. H-167. PMID:11086690

  10. Precursor directed biosynthesis of odd-numbered fatty acids by different yeasts.

    PubMed

    Řezanka, Tomáš; Kolouchová, Irena; Sigler, Karel

    2015-09-01

    Precursor-directed biosynthesis was used for directed preparation of positional isomers of heptadecanoic acid (17:1), which have convenient pharmacological properties. Cultivation of Candida sp., Kluyveromyces polysporus, Rhodotorula glutinis, Saccharomyces cerevisiae, Torulaspora delbrueckii, Trichosporon cutaneum, and Yarrowia lipolytica on 20 g/L glucose, 4 g/L acetic, or 4 g/L propionic acids yielded different proportions of 17:1. Cultivation on carbon sources with even numbers of carbon atoms (glucose and acetic acid) produced preferentially 8Z- and 10Z-heptadecenoic acids in about equal amounts, in agreement with the proposed biosynthesis of fatty acids, whereas cultivation on propionic acid as the only carbon source produced over 90 % of total fatty acids of 9-17:1 out of all possible positional isomers. The structures of positional isomers of 17:1 acid were determined using dimethyl disulfides of fatty acid methyl esters. In cultivation of Candida sp. on propionic acid, the yield of heptadecenoic acid reached 111 mg/L cultivation medium. Principal component analysis was used for identifying the effect of cultivation conditions on the production of the 17:1 acid by individual yeast strains. PMID:25813199

  11. Effect of cryoprotectants on the survival of cascudo preto (Rhinelepis aspera) embryos stored at -8 °C.

    PubMed

    Fornari, Darci Carlos; Ribeiro, Ricardo Pereira; Streit, Danilo; Godoy, Leandro Cesar; Neves, Patrícia Ribeiro; de Oliveira, Diego; Sirol, Rodolfo Nardez

    2014-02-01

    Cryopreservation of germplasm provides a promising method to preserve fish genetic material, which is of great importance in preservation of species diversity, aquaculture, and management of fish models used in biomedical research. In the present study, cryopreservation of Rhinelepis aspera embryos, a Brazilian endangered species, was studied for the first time using a short-term cooling protocol. Embryos at blastoporous closing stage were selected, placed in 6-ml glass vials and stored at -8 °C for 6 h in 10 different cryoprotectant solutions: S1 (17.1% sucrose + 9% methanol); S2 (17.1% sucrose + 9% DMSO); S3 (8.5% sucrose + 8.5% glucose + 9% methanol); S4 (8.5% sucrose + 8.5% glucose + 9% DMSO); S5 (17.1% sucrose + 9% ethylene glycol); S6 (8.5% sucrose + 8.5% glucose + 9% ethylene glycol); S7 (17.1% sucrose + 4.5% methanol + 4.5% DMSO); S8 (17.1% sucrose + 4.5% methanol + 4.5% ethylene glycol); S9 (17.1% sucrose + 4.5% DMSO + 4.5% ethylene glycol); and S10 (100% water). Embryo viability was assessed by hatching rate, counting live larvae and number of failed eggs under a stereomicroscope. The results showed that only the cryoprotectant solutions that contained methanol associated to sucrose (S1, S7 and S8) provided partial protection of Rhinelepis aspera embryos from cold damage (over 50% hatching rate in S1), while the use of DMSO and ethylene glycol, isolated or in combination, resulted in no hatching rate. Further studies are needed in order to extend the storage time and to improve the hatching rate for the species.

  12. Antifouling Compounds from the Marine-Derived Fungus Aspergillus terreus SCSGAF0162.

    PubMed

    Nong, Xu-Hua; Zhang, Xiao-Yong; Xu, Xin-Ya; Qi, Shu-Hua

    2015-06-01

    A new cyclic tetrapeptide, asperterrestide B (1), and 11 known compounds (2-12) were isolated from a marine-derived fungus Aspergillus terreus SCSGAF0162. The structure of 1 was elucidated by spectroscopic analysis, and the absolute configuration of 1 was determined by Mosher ester and Marfey's methods. Compounds 4, 6, and 8 had potent antifouling activity against larvae of the barnacle Balanus amphitrite, with EC50 values of 17.1 ± 1.2, 11.6 ± 0.6, and 17.1 ± 0.8 μg x mL(-1), respectively. PMID:26197544

  13. Effects of copper-plasma deposition on weathering properties of wood surfaces

    NASA Astrophysics Data System (ADS)

    Gascón-Garrido, P.; Mainusch, N.; Militz, H.; Viöl, W.; Mai, C.

    2016-03-01

    Thin layers of copper micro-particles were deposited on the surfaces of Scots pine (Pinus sylvestris L.) micro-veneers using atmospheric pressure plasma to improve the resistance of the surfaces to weathering. Three different loadings of copper were established. Micro-veneers were exposed to artificial weathering in a QUV weathering tester for 0, 24, 48, 96 and 144 h following the standard EN 927-6 [1]. Mass losses after each exposure showed significant differences between copper coated and untreated micro-veneers. Tensile strength was assessed at zero span (z-strength) and finite span (f-strength) under dry conditions (20 °C, 65% RH). During 48 h, micro-veneers lost their z-strength progressively. In contrast, copper coating at highest loading imparts a photo-protective effect to wood micro-veneers during 144 h exhibiting z-strength retention of 95%. F-strength losses were similar in all copper treated and untreated micro-veneers up to 96 h. However, after 144 h, copper coated micro-veneers at highest loading showed significantly greater strength retention of 56%, while untreated micro-veneers exhibited only 38%. Infrared spectroscopy suggested that copper coating does not stabilize lignin. Inductively Coupled Plasma revealed that micro-veneers coated with the highest loading exhibited the lowest percentage of copper loss. Blue stain resistance of copper coated Scots pine following the guidelines of EN 152 [2] was performed. Additional test with different position of the coated surface was also assessed. Copper coating reduced fungal growth when coated surface is exposed in contact with vermiculite. Spores of Aureobasidium pullulans were not able to germinate on the copper coated surface positioned uppermost.

  14. Beta-Amylases from Alfalfa (Medicago sativa L.) Roots 1

    PubMed Central

    Doehlert, Douglas C.; Duke, Stanley H.; Anderson, Laurens

    1982-01-01

    Amylase was found in high activity (193 international units per milligram protein) in the tap root of alfalfa (Medicago sativa L. cv. Sonora). The activity was separated by gel filtration chromatography into two fractions with molecular weights of 65,700 (heavy amylase) and 41,700 (light amylase). Activity staining of electrophoretic gels indicated the presence of one isozyme in the heavy amylase fraction and two in the light amylase fraction. Three amylase isozymes with electrophoretic mobilities identical to those in the heavy and the light amylase fractions were the only amylases identified in crude root preparations. Both heavy and light amylases hydrolyzed amylopectin, soluble starch, and amylose but did not hydrolyze pullulan or β-limit dextrin. The ratio of viscosity change to reducing power production during starch hydrolysis was identical for both alfalfa amylase fractions and sweet potato β-amylase, while that of bacterial α-amylase was considerably higher. The identification of maltose and β-limit dextrin as hydrolytic end-products confirmed that these alfalfa root amylases are all β-amylases. The pH optimum for both β-amylase fractions was 6.0. Both light and heavy β-amylases showed normal Michaelis-Menten kinetics, with soluble starch as substrate, and had respectively Km values of 5.9 and 6.8 milligrams starch per milliliter and Vmax of 640 and 130 international units per milligram protein. Arrhenius plots indicated that the energy of activation for the heavy β-amylase remained relatively unchanged (12.7 to 13.0 kilocalories per mole) from 0 to 30°C, whereas the energy of activation for the light amylase increased from 12.0 to about 28.0 kilocalories per mole at 8.7°C as temperature was lowered. The light amylase was shown to be inhibited by maltose. Images PMID:16662350

  15. [Concentration of allergic fungi spores in the air of flats in Lódź].

    PubMed

    Krawczyk, P; Kowalski, M L; Ochecka-Szymańska, A

    1999-01-01

    The real contribution of moulds to the pathogenesis of allergic diseases remains unknown, although positive skin prick tests and/or specific serum IgE to moki allergens can be detected in 1-5% of atopic patients. A significant problem in assesment of exposure to mould allergens, resulting with difficulty in standarization of methods. The aim of this work was to assess the concentration of spores of 8 mould species in flats inhabited by peoples who Bont show any symptoms of allergy. The Open Petri Dish (OPD) method involving sedimentation of participles contained in the column of air over the dish was used to assess the number of spores in 1 m3 of indoor atmospheres. All colonies were counted, but only 8 mould species implicated in inhaled allergy were identified, ie.: Alternaria tenuis, Cladosporium herbarum, Helminthosporum halodes, Pullularia pullulans, Penicillium notatam, Rhizopus nigricans, Mucor mucedo, Aspergillus fumigatus. The tests were carried out in 10 flats located in various quarters of the cify of Lodź during three consecutive days of September 1995 between 5:00 pm and 6:04 pm. In analyzing the percentage of spores of each of the eight mould species tested we determined that, independent of fiat and test day, C. herbarum predominated. It is good agreement with the observations of other authors who report that among large quantities of fungi that are detected in late summer, usually C. herbarum spores dominate. This is the season when the incidence of the Cladosporium spores in the atmospheric air increases. Spores of H. halodes were detected least frequently. Our study demonstrated the presence of substantial amounts of mould spores in indoor air of houses in Lódź. The spores belong to species with documented allergenicity, suggesting that they may play a role in development of allergic sensitization in susceptible subjects.

  16. 240 Mold Sensitization in Chronic Rhinosinusitis Patients

    PubMed Central

    Gawlik, Radoslaw; Czecior, Eugeniusz

    2012-01-01

    Background It is estimated that about 10% of the population have IgE antibodies to common inhalant molds. Exposure to fungal allergens could be linked to the presence and persistence of asthma, rhinitis and atopic dermatitis. Mold sensitization is a risk factor for development and deterioration of upper airway allergy, especially chronic rhinosinusitis. We addressed the incidence of mold allergy measured as specific IgE to molds and skin prick tests in chronic sinusitis patients. We assessed prevalence of allergic reactions to mould among surgery treated chronic sinusitis patients. Methods A group of 28 chronic sinusitis patients after surgery were included into the study. Routine medical examination, skin prick tests with common inhaled allergens and extended mold panel (Alternaria alternate, Cladosporium herbarium, Aspergilus fumigatus, Candida albicans, Mucor mucedo, Botrytis cinerea, Rhisopus nigricans, Penicilliumi notatum, Fusarum moniliforme Pullularia pullulans (Allergopharma, Germany), tIgE, asIgE measurement were performed (Phadia, Sweden). All investigated patients were consulted by laryngologist and mycological examination was performed. Results We found that sensitization to at least one allergen was present in 43.8(14/32) of sinusitis patients. The most prevalent was sensitization to house dust mite Dermatophagoides pt., found in 21.8 % (7/32) patients. Positive results of skin prick tests with Candida albicans we observed in 18.8% (6/32), with Alternaria alternate in 15,6% (5/32), Cladosporium herbarium in 6,3% (2/32), Aspergilus fumigatus in 3,13 % (1/32). None of investigated patients presented sensitization to other mold allergens. Microbiological methods demonstrated fungal infection only in 2 patients. Conclusions Almost half of chronic sinusitis patients presented sensitization to at least one allergen. Fungal allergy is relatively rare in chronic sinusitis patients.

  17. Wetting of biopolymer coatings: contact angle kinetics and image analysis investigation.

    PubMed

    Farris, Stefano; Introzzi, Laura; Biagioni, Paolo; Holz, Torsten; Schiraldi, Alberto; Piergiovanni, Luciano

    2011-06-21

    The surface wetting of five biopolymers, used as coating materials for a plastic film, was monitored over a span of 8 min by means of the optical contact angle technique. Because most of the total variation was observed to occur during the first 60 s, we decided to focus on this curtailed temporal window. Initial contact angle values (θ(0)) ranged from ∼91° for chitosan to ∼30° for pullulan. However, the water drop profile began to change immediately following drop deposition for all biocoatings, confirming that the concept of water contact angle equilibrium is not applicable to most biopolymers. First, a three-parameter decay equation [θ(t) = θ(0) exp(kt(n))] was fit to the experimental contact angle data to describe the kinetics of the contact angle change for each biocoating. Interestingly, the k constant correlated well with the contact angle evolution rate and the n exponent seemed to be somehow linked to the physicochemical phenomena underlying the overall kinetics process. Second, to achieve a reliable description of droplet evolution, the contact angle (CA) analysis was coupled with image analysis (IA) through a combined geometric/trigonometric approach. Absorption and spreading were the key factors governing the overall mechanism of surface wetting during the 60 s analysis, although the individual quantification of both phenomena demonstrated that spreading provided the largest contribution for all biopolymers, with the only exception of gelatin, which showed two quasi-equivalent and counterbalancing effects. The possible correlation between these two phenomena and the topography of the biopolymer surfaces are then discussed on the basis of atomic force microscopy analyses. PMID:21619017

  18. Systems Biology of Microbial Exopolysaccharides Production

    PubMed Central

    Ates, Ozlem

    2015-01-01

    Exopolysaccharides (EPSs) produced by diverse group of microbial systems are rapidly emerging as new and industrially important biomaterials. Due to their unique and complex chemical structures and many interesting physicochemical and rheological properties with novel functionality, the microbial EPSs find wide range of commercial applications in various fields of the economy such as food, feed, packaging, chemical, textile, cosmetics and pharmaceutical industry, agriculture, and medicine. EPSs are mainly associated with high-value applications, and they have received considerable research attention over recent decades with their biocompatibility, biodegradability, and both environmental and human compatibility. However, only a few microbial EPSs have achieved to be used commercially due to their high production costs. The emerging need to overcome economic hurdles and the increasing significance of microbial EPSs in industrial and medical biotechnology call for the elucidation of the interrelations between metabolic pathways and EPS biosynthesis mechanism in order to control and hence enhance its microbial productivity. Moreover, a better understanding of biosynthesis mechanism is a significant issue for improvement of product quality and properties and also for the design of novel strains. Therefore, a systems-based approach constitutes an important step toward understanding the interplay between metabolism and EPS biosynthesis and further enhances its metabolic performance for industrial application. In this review, primarily the microbial EPSs, their biosynthesis mechanism, and important factors for their production will be discussed. After this brief introduction, recent literature on the application of omics technologies and systems biology tools for the improvement of production yields will be critically evaluated. Special focus will be given to EPSs with high market value such as xanthan, levan, pullulan, and dextran. PMID:26734603

  19. Sequence-based analysis of the Vitis vinifera L. cv Cabernet Sauvignon grape must mycobiome in three South African vineyards employing distinct agronomic systems

    SciTech Connect

    Setati, Mathabatha E.; Jacobson, Daniel; Bauer, Florian F.

    2015-11-30

    Recent microbiomic research of agricultural habitats has highlighted tremendous microbial biodiversity associated with such ecosystems. In addition, data generated in vineyards have furthermore highlighted significant regional differences in vineyard biodiversity, hinting at the possibility that such differences might be responsible for regional differences in wine style and character, a hypothesis referred to as "microbial terroir." The current study further contributes to this body of work by comparing the mycobiome associated with South African (SA) Cabernet Sauvignon grapes in three neighboring vineyards that employ different agronomic approaches, and comparing the outcome with similar data sets from Californian vineyards. The aim of this study was to fully characterize the mycobiomes associated with the grapes from these vineyards. The data revealed approximately 10 times more fungal diversity than what is typically retrieved from culture-based studies. The Biodynamic vineyard was found to harbor a more diverse fungal community (H = 2.6) than the conventional (H = 2.1) and integrated (H = 1.8) vineyards. The data show that ascomycota are the most abundant phylum in the three vineyards, with Aureobasidium pullulans and its close relative Kabatiella microsticta being the most dominant fungi. This is the first report to reveal a high incidence of K. microsticta in the grape/wine ecosystem. Different common wine yeast species, such as Metschnikowia pulcherrima and Starmerella bacillaris dominated the mycobiome in the three vineyards. The data show that the filamentous fungi are the most abundant community in grape must although they are not regarded as relevant during wine fermentation. Comparison of metagenomic datasets from the three SA vineyards and previously published data from Californian vineyards revealed only 25% of the fungi in the SA dataset was also present in the Californian dataset, with greater variation evident amongst

  20. Sequence-based analysis of the Vitis vinifera L. cv Cabernet Sauvignon grape must mycobiome in three South African vineyards employing distinct agronomic systems

    DOE PAGESBeta

    Setati, Mathabatha E.; Jacobson, Daniel; Bauer, Florian F.

    2015-11-30

    Recent microbiomic research of agricultural habitats has highlighted tremendous microbial biodiversity associated with such ecosystems. In addition, data generated in vineyards have furthermore highlighted significant regional differences in vineyard biodiversity, hinting at the possibility that such differences might be responsible for regional differences in wine style and character, a hypothesis referred to as "microbial terroir." The current study further contributes to this body of work by comparing the mycobiome associated with South African (SA) Cabernet Sauvignon grapes in three neighboring vineyards that employ different agronomic approaches, and comparing the outcome with similar data sets from Californian vineyards. The aim ofmore » this study was to fully characterize the mycobiomes associated with the grapes from these vineyards. The data revealed approximately 10 times more fungal diversity than what is typically retrieved from culture-based studies. The Biodynamic vineyard was found to harbor a more diverse fungal community (H = 2.6) than the conventional (H = 2.1) and integrated (H = 1.8) vineyards. The data show that ascomycota are the most abundant phylum in the three vineyards, with Aureobasidium pullulans and its close relative Kabatiella microsticta being the most dominant fungi. This is the first report to reveal a high incidence of K. microsticta in the grape/wine ecosystem. Different common wine yeast species, such as Metschnikowia pulcherrima and Starmerella bacillaris dominated the mycobiome in the three vineyards. The data show that the filamentous fungi are the most abundant community in grape must although they are not regarded as relevant during wine fermentation. Comparison of metagenomic datasets from the three SA vineyards and previously published data from Californian vineyards revealed only 25% of the fungi in the SA dataset was also present in the Californian dataset, with greater variation evident amongst ubiquitous epiphytic

  1. Molar mass characterization of sodium carboxymethyl cellulose by SEC-MALLS.

    PubMed

    Shakun, Maryia; Maier, Helena; Heinze, Thomas; Kilz, Peter; Radke, Wolfgang

    2013-06-01

    Two series of sodium carboxymethyl celluloses (NaCMCs) derived from microcrystalline cellulose (Avicel samples) and cotton linters (BWL samples) with average degrees of substitution (DS) ranging from DS=0.45 to DS=1.55 were characterized by size exclusion chromatography with multi-angle laser light scattering detection (SEC-MALLS) in 100 mmol/L aqueous ammonium acetate (NH4OAc) as vaporizable eluent system. The application of vaporizable NH4OAc allows future use of the eluent system in two-dimensional separations employing evaporative light scattering detection (ELSD). The losses of samples during filtration and during the chromatographic experiment were determined. The scaling exponent as of the relation [Formula: see text] was approx. 0.61, showing that NaCMCs exhibit an expanded coil conformation in solution. No systematic dependencies of as on DS were observed. The dependences of molar mass on SEC-elution volume for samples of different DS can be well described by a common calibration curve, which is of advantage, as it allows the determination of molar masses of unknown samples by using the same calibration curve, irrespective of the DS of the NaCMC sample. Since no commercial NaCMC standards are available, correction factors were determined allowing converting a pullulan based calibration curve into a NaCMC calibration using the broad calibration approach. The weight average molar masses derived using the so established calibration curve closely agree with the ones determined by light scattering, proving the accuracy of the correction factors determined.

  2. Purification and characterization of the extracellular. alpha. -amylase from Clostridium acetobutylicum ATCC 824

    SciTech Connect

    Paquet, V.; Croux, C.; Goma, G.; Soucaille, P. )

    1991-01-01

    The extracellular {alpha}-amylase (1,4-{alpha}-D-glucanglucanohydrolase; EC 3.2.1.1) from Clostridium acetobutylicum ATCC 824 was purified to homogeneity by anion-exchange chromatography (Mono Q) and gel filtration (Superose 12). The enzyme had an isoelectric point of 4.7 and a molecular weight of 84,000, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. It was a monomeric protein, the 19-amino-acid N terminus of which displayed 42% homology with the Bacillus subtilis saccharifying {alpha}-amylase. The amino acid composition of the enzyme showed a high number of acidic and hydrophobic residues and only one cysteine residue per mole. The activity of the {alpha}-amylase was not stimulated by calcium ions (or other metal ions) or inhibited by EDTA, although the enzyme contained seven calcium atoms per molecule. {alpha}-Amylase activity on soluble starch was optimal at pH 5.6 and 45{degree}C. The {alpha}-amylase was stable at an acidic pH but very sensitive to thermal inactivation. It hydrolyzed soluble starch, with a K{sub m} of 3.6 g {center dot} liter{sup {minus}1} and a K{sub cat} of 122 mol of reducing sugars {center dot} s{sup {minus}1} {center dot} mol{sup {minus}1}. The {alpha}-amylase showed greater activity with high-molecular-weight substrates than with low-molecular-weight maltooligosaccharides, hydrolyzed glycogen and pullulan slowly, but did not hydrolyze dextran or cyclodextrins. The major end products of maltohexaose degradation were glucose, maltose, and maltotriose; maltotetraose and maltopentaose were formed as intermediate products. Twenty seven percent of the glucoamylase activity generally detected in the culture supernatant of C. acetobutylicum can be attributed to the {alpha}-amylase.

  3. Terbinafine hydrochloride loaded liposome film formulation for treatment of onychomycosis: in vitro and in vivo evaluation.

    PubMed

    Tanrıverdi, Sakine Tuncay; Hilmioğlu Polat, Süleyha; Yeşim Metin, Dilek; Kandiloğlu, Gülşen; Özer, Özgen

    2016-01-01

    Onychomycosis is a fungal infection of nail unit that is caused by dermatophytes. Oral Terbinafine hydrochloride (TBF-HCl) is being used for the treatment of onychomycosis since 24 years. The side effects caused by the systemic application and limitations of topical administration of this drug regarding the diffusion through nail lead to the development of a new formulation based on, TBF-HCl-loaded liposome. The newly obtained film formulations were prepared and characterized via several parameters, such as physical appearance, drug content, thickness, bioadhesive properties and tensile strength. In vitro and ex vivo permeation studies were performed to select an optimum film formulation for antifungal activity to show the efficiency of formulations regarding the treatment of onychomycosis. The in vitro release percentages of drug were found 71.6 ± 3.28, 54.4 ± 4.26, 56.1 ± 7.48 and 46.0 ± 2.43 for liposome loaded pullulan films (LI-P, LII-P) and liposome loaded Eudragit films (LI-E, LII-E), respectively. The accumulated drug in the nail plates were found 31.16 ± 4.22, 24.81 ± 5.35, 8.17 ± 1.81 and 8.92 ± 3.37 for LI-P, LII-P, LI-E and LII-E, respectively, which within therapeutic range for all film formulations. The accumulated drug in the nail plate was found within therapeutic range for all film formulations. The efficacy of the selected TBF-HCl-loaded liposome film formulation was compared with TBF-HCl-loaded liposome, ethosome, liposome poloxamer gel and ethosome chitosan gel formulations. It was found that TBF-HCl-loaded liposome film formulation had better antifungal activity on fungal nails which make this liposome film formulation promising for ungual therapy of fungal nail infection.

  4. Sequence-based Analysis of the Vitis vinifera L. cv Cabernet Sauvignon Grape Must Mycobiome in Three South African Vineyards Employing Distinct Agronomic Systems

    PubMed Central

    Setati, Mathabatha E.; Jacobson, Daniel; Bauer, Florian F.

    2015-01-01

    Recent microbiomic research of agricultural habitats has highlighted tremendous microbial biodiversity associated with such ecosystems. Data generated in vineyards have furthermore highlighted significant regional differences in vineyard biodiversity, hinting at the possibility that such differences might be responsible for regional differences in wine style and character, a hypothesis referred to as “microbial terroir.” The current study further contributes to this body of work by comparing the mycobiome associated with South African (SA) Cabernet Sauvignon grapes in three neighboring vineyards that employ different agronomic approaches, and comparing the outcome with similar data sets from Californian vineyards. The aim of this study was to fully characterize the mycobiomes associated with the grapes from these vineyards. The data revealed approximately 10 times more fungal diversity than what is typically retrieved from culture-based studies. The Biodynamic vineyard was found to harbor a more diverse fungal community (H = 2.6) than the conventional (H = 2.1) and integrated (H = 1.8) vineyards. The data show that ascomycota are the most abundant phylum in the three vineyards, with Aureobasidium pullulans and its close relative Kabatiella microsticta being the most dominant fungi. This is the first report to reveal a high incidence of K. microsticta in the grape/wine ecosystem. Different common wine yeast species, such as Metschnikowia pulcherrima and Starmerella bacillaris dominated the mycobiome in the three vineyards. The data show that the filamentous fungi are the most abundant community in grape must although they are not regarded as relevant during wine fermentation. Comparison of metagenomic datasets from the three SA vineyards and previously published data from Californian vineyards revealed only 25% of the fungi in the SA dataset was also present in the Californian dataset, with greater variation evident amongst ubiquitous epiphytic fungi. PMID

  5. Disruption of a sugar transporter gene cluster in a hyperthermophilic archaeon using a host-marker system based on antibiotic resistance.

    PubMed

    Matsumi, Rie; Manabe, Kenji; Fukui, Toshiaki; Atomi, Haruyuki; Imanaka, Tadayuki

    2007-04-01

    We have developed a gene disruption system in the hyperthermophilic archaeon Thermococcus kodakaraensis using the antibiotic simvastatin and a fusion gene designed to overexpress the 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase gene (hmg(Tk)) with the glutamate dehydrogenase promoter. With this system, we disrupted the T. kodakaraensis amylopullulanase gene (apu(Tk)) or a gene cluster which includes apu(Tk) and genes encoding components of a putative sugar transporter. Disruption plasmids were introduced into wild-type T. kodakaraensis KOD1 cells, and transformants exhibiting resistance to 4 microM simvastatin were isolated. The transformants exhibited growth in the presence of 20 microM simvastatin, and we observed a 30-fold increase in intracellular HMG-CoA reductase activity. The expected gene disruption via double-crossover recombination occurred at the target locus, but we also observed recombination events at the hmg(Tk) locus when the endogenous hmg(Tk) gene was used. This could be avoided by using the corresponding gene from Pyrococcus furiosus (hmg(Pf)) or by linearizing the plasmid prior to transformation. While both gene disruption strains displayed normal growth on amino acids or pyruvate, cells without the sugar transporter genes could not grow on maltooligosaccharides or polysaccharides, indicating that the gene cluster encodes the only sugar transporter involved in the uptake of these compounds. The Deltaapu(Tk) strain could not grow on pullulan and displayed only low levels of growth on amylose, suggesting that Apu(Tk) is a major polysaccharide-degrading enzyme in T. kodakaraensis.

  6. Present and future medical applications of microbial exopolysaccharides

    PubMed Central

    Moscovici, Misu

    2015-01-01

    Microbial exopolysaccharides (EPS) have found outstanding medical applications since the mid-20th century, with the first clinical trials on dextran solutions as plasma expanders. Other EPS entered medicine firstly as conventional pharmaceutical excipients (e.g., xanthan – as suspension stabilizer, or pullulan – in capsules and oral care products). Polysaccharides, initially obtained from plant or animal sources, became easily available for a wide range of applications, especially when they were commercially produced by microbial fermentation. Alginates are used as anti-reflux, dental impressions, or as matrix for tablets. Hyaluronic acid and derivatives are used in surgery, arthritis treatment, or wound healing. Bacterial cellulose is applied in wound dressings or scaffolds for tissue engineering. The development of drug controlled-release systems and of micro- and nanoparticulated ones, has opened a new era of medical applications for biopolymers. EPS and their derivatives are well-suited potentially non-toxic, biodegradable drug carriers. Such systems concern rating and targeting of controlled release. Their large area of applications is explained by the available manifold series of derivatives, whose useful properties can be thereby controlled. From matrix inclusion to conjugates, different systems have been designed to solubilize, and to assure stable transport in the body, target accumulation and variable rate-release of a drug substance. From controlled drug delivery, EPS potential applications expanded to vaccine adjuvants and diagnostic imaging systems. Other potential applications are related to the bioactive (immunomodulator, antitumor, antiviral) characteristics of EPS. The numerous potential applications still wait to be developed into commercial pharmaceuticals and medical devices. Based on previous and recent results in important medical-pharmaceutical domains, one can undoubtedly state that EPS medical applications have a broad future ahead

  7. Sustained Ocular Delivery of Ciprofloxacin Using Nanospheres and Conventional Contact Lens Materials

    PubMed Central

    Garhwal, Rahul; Shady, Sally F.; Ellis, Edward J.; Ellis, Jeanne Y.; Leahy, Charles D.; McCarthy, Stephen P.; Crawford, Kathryn S.

    2012-01-01

    Purpose. To formulate conventional contact lenses that incorporate nanosphere-encapsulated antibiotic and demonstrate that the lenses provide for sustained antibacterial activity. Methods. A copolymer composed of pullulan and polycaprolactone (PCL) was used to synthesize core-shell nanospheres that encapsulated ciprofloxacin. Bactericidal activity of the nanosphere-encapsulated ciprofloxacin (nanosphere/cipro) was tested by using liquid cultures of either Staphylococcus aureus or Pseudomonas aeruginosa. Nanosphere/cipro was then incorporated into HEMA-based contact lenses that were tested for growth inhibition of S. aureus or P. aeruginosa in liquid cultures inoculated daily with fresh bacteria. Lens designs included thin or thick lenses incorporating nanosphere/cipro and ciprofloxacin-HCl-soaked Acuvue lenses (Acuvue; Johnson & Johnson Vision Care, Inc., Jacksonville, FL). Results. Less than 2 μg/mL of nanosphere/cipro effectively inhibited the proliferation of cultures inoculated with 107 or 108 bacteria/mL of S. aureus and P. aeruginosa, respectively. HEMA-based contact lenses polymerized with nanosphere/cipro were transparent, effectively inhibited the proliferation of greater than 107/mL of bacteria added daily over 3 days of culture, and killed up to 5 × 109 total microbes in a single inoculation. A thicker lens design provided additional inhibition of bacterial growth for up to 96 hours. Conclusions. Core-shell nanospheres loaded with an antibiotic can be incorporated into a conventional, transparent contact lens and provide for sustained and effective bactericidal activity and thereby provide a new drug delivery platform for widespread use in treating ocular disorders. PMID:22266514

  8. Are Epiphytic Microbial Communities in the Carposphere of Ripening Grape Clusters (Vitis vinifera L.) Different between Conventional, Organic, and Biodynamic Grapes?

    PubMed Central

    Kecskeméti, Elizabeth; Berkelmann-Löhnertz, Beate; Reineke, Annette

    2016-01-01

    Using barcoded pyrosequencing fungal and bacterial communities associated with grape berry clusters (Vitis vinifera L.) obtained from conventional, organic and biodynamic vineyard plots were investigated in two subsequent years at different stages during berry ripening. The four most abundant operational taxonomic units (OTUs) based on fungal ITS data were Botrytis cinerea, Cladosporium spp., Aureobasidium pullulans and Alternaria alternata which represented 57% and 47% of the total reads in 2010 and 2011, respectively. Members of the genera Sphingomonas, Gluconobacter, Pseudomonas, Erwinia, and Massilia constituted 67% of the total number of bacterial 16S DNA reads in 2010 samples and 78% in 2011 samples. Viticultural management system had no significant effect on abundance of fungi or bacteria in both years and at all three sampling dates. Exceptions were A. alternata and Pseudomonas spp. which were more abundant in the carposphere of conventional compared to biodynamic berries, as well as Sphingomonas spp. which was significantly less abundant on conventional compared to organic berries at an early ripening stage in 2011. In general, there were no significant differences in fungal and bacterial diversity indices or richness evident between management systems. No distinct fungal or bacterial communities were associated with the different maturation stages or management systems, respectively. An exception was the last stage of berry maturation in 2011, where the Simpson diversity index was significantly higher for fungal communities on biodynamic compared to conventional grapes. Our study highlights the existence of complex and dynamic microbial communities in the grape cluster carposphere including both phytopathogenic and potentially antagonistic microorganisms that can have a significant impact on grape production. Such knowledge is particularly relevant for development, selection and application of effective control measures against economically important

  9. Yeast β-1,6-glucan is a primary target for the Saccharomyces cerevisiae K2 toxin.

    PubMed

    Lukša, Juliana; Podoliankaitė, Monika; Vepštaitė, Iglė; Strazdaitė-Žielienė, Živilė; Urbonavičius, Jaunius; Servienė, Elena

    2015-04-01

    Certain Saccharomyces cerevisiae strains secrete different killer proteins of double-stranded-RNA origin. These proteins confer a growth advantage to their host by increasing its survival. K2 toxin affects the target cell by binding to the cell surface, disrupting the plasma membrane integrity, and inducing ion leakage. In this study, we determined that K2 toxin saturates the yeast cell surface receptors in 10 min. The apparent amount of K2 toxin, bound to a single cell of wild type yeast under saturating conditions, was estimated to be 435 to 460 molecules. It was found that an increased level of β-1,6-glucan directly correlates with the number of toxin molecules bound, thereby impacting the morphology and determining the fate of the yeast cell. We observed that the binding of K2 toxin to the yeast surface receptors proceeds in a similar manner as in case of the related K1 killer protein. It was demonstrated that the externally supplied pustulan, a poly-β-1,6-glucan, but not the glucans bearing other linkage types (such as laminarin, chitin, and pullulan) efficiently inhibits the K2 toxin killing activity. In addition, the analysis of toxin binding to the intact cells and spheroplasts confirmed that majority of K2 protein molecules attach to the β-1,6-glucan, rather than the plasma membrane-localized receptors. Taken together, our results reveal that β-1,6-glucan is a primary target of K2 toxin and is important for the execution of its killing property.

  10. Associations between Fungal Species and Water-Damaged Building Materials ▿

    PubMed Central

    Andersen, Birgitte; Frisvad, Jens C.; Søndergaard, Ib; Rasmussen, Ib S.; Larsen, Lisbeth S.

    2011-01-01

    Fungal growth in damp or water-damaged buildings worldwide is an increasing problem, which has adverse effects on both the occupants and the buildings. Air sampling alone in moldy buildings does not reveal the full diversity of fungal species growing on building materials. One aim of this study was to estimate the qualitative and quantitative diversity of fungi growing on damp or water-damaged building materials. Another was to determine if associations exist between the most commonly found fungal species and different types of materials. More than 5,300 surface samples were taken by means of V8 contact plates from materials with visible fungal growth. Fungal identifications and information on building material components were analyzed using multivariate statistic methods to determine associations between fungi and material components. The results confirmed that Penicillium chrysogenum and Aspergillus versicolor are the most common fungal species in water-damaged buildings. The results also showed Chaetomium spp., Acremonium spp., and Ulocladium spp. to be very common on damp building materials. Analyses show that associated mycobiotas exist on different building materials. Associations were found between (i) Acremonium spp., Penicillium chrysogenum, Stachybotrys spp., Ulocladium spp., and gypsum and wallpaper, (ii) Arthrinium phaeospermum, Aureobasidium pullulans, Cladosporium herbarum, Trichoderma spp., yeasts, and different types of wood and plywood, and (iii) Aspergillus fumigatus, Aspergillus melleus, Aspergillus niger, Aspergillus ochraceus, Chaetomium spp., Mucor racemosus, Mucor spinosus, and concrete and other floor-related materials. These results can be used to develop new and resistant building materials and relevant allergen extracts and to help focus research on relevant mycotoxins, microbial volatile organic compounds (MVOCs), and microparticles released into the indoor environment. PMID:21531835

  11. Diversity and antifungal activity of the endophytic fungi associated with the native medicinal cactus Opuntia humifusa (Cactaceae) from the United States.

    PubMed

    Silva-Hughes, Alice F; Wedge, David E; Cantrell, Charles L; Carvalho, Camila R; Pan, Zhiqiang; Moraes, Rita M; Madoxx, Victor L; Rosa, Luiz H

    2015-06-01

    The endophytic fungal community associated with the native cactus Opuntia humifusa in the United States was investigated and its potential for providing antifungal compounds. A hundred-eight endophytic fungal isolates were obtained and identified by molecular methods into 17 different taxa of the genera Alternaria, Aureobasidium, Biscogniauxia, Cladosporium, Cryptococcus, Curvularia, Diaporthe, Epicoccum, Paraconiothyrium, Pestalotiopsis and Phoma. The most frequent species associated with O. humifusa were Alternaria sp. 3, Aureobasidium pullulans and Diaporthe sp. The fungal community of O. humifusa had a high richness and diversity; additionally, the species richness obtained indicates that the sample effort was enough to recover the diversity pattern obtained. Six extracts of endophytes showed antifungal properties and (1)H NMR analyses of the extracts of Alternaria sp. 5 Ohu 8B2, Alternaria sp. 3 Ohu 30A, Cladosporium funiculosum Ohu 17C1 and Paraconiothyrium sp. Ohu 17A indicated the presence of functional groups associated with unsaturated fatty-acid olefinic protons and fatty acid methylene and methyl protons. GC-FID analysis of these extracts confirmed the presence of a mixture of different fatty acids. The (1)H NMR analyses of Biscogniauxia mediterranea Ohu 19B extracts showed the presence of aromatic compounds. From the extract of B. mediterranea we isolated the compound 5-methylmellein that displayed moderate antifungal activity against the phytopathogenic fungi Phomopsis obscurans. Our results suggest that native medicinal cacti of the United States can live symbiotically with rich and diverse endophytic communities and may be a source of bioactive molecules, including those able to inhibit or control plant disease pathogens.

  12. Disruption of a sugar transporter gene cluster in a hyperthermophilic archaeon using a host-marker system based on antibiotic resistance.

    PubMed

    Matsumi, Rie; Manabe, Kenji; Fukui, Toshiaki; Atomi, Haruyuki; Imanaka, Tadayuki

    2007-04-01

    We have developed a gene disruption system in the hyperthermophilic archaeon Thermococcus kodakaraensis using the antibiotic simvastatin and a fusion gene designed to overexpress the 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase gene (hmg(Tk)) with the glutamate dehydrogenase promoter. With this system, we disrupted the T. kodakaraensis amylopullulanase gene (apu(Tk)) or a gene cluster which includes apu(Tk) and genes encoding components of a putative sugar transporter. Disruption plasmids were introduced into wild-type T. kodakaraensis KOD1 cells, and transformants exhibiting resistance to 4 microM simvastatin were isolated. The transformants exhibited growth in the presence of 20 microM simvastatin, and we observed a 30-fold increase in intracellular HMG-CoA reductase activity. The expected gene disruption via double-crossover recombination occurred at the target locus, but we also observed recombination events at the hmg(Tk) locus when the endogenous hmg(Tk) gene was used. This could be avoided by using the corresponding gene from Pyrococcus furiosus (hmg(Pf)) or by linearizing the plasmid prior to transformation. While both gene disruption strains displayed normal growth on amino acids or pyruvate, cells without the sugar transporter genes could not grow on maltooligosaccharides or polysaccharides, indicating that the gene cluster encodes the only sugar transporter involved in the uptake of these compounds. The Deltaapu(Tk) strain could not grow on pullulan and displayed only low levels of growth on amylose, suggesting that Apu(Tk) is a major polysaccharide-degrading enzyme in T. kodakaraensis. PMID:17259314

  13. Plant growth-promoting and antifungal activity of yeasts from dark chestnut soil.

    PubMed

    Ignatova, Lyudmila V; Brazhnikova, Yelena V; Berzhanova, Ramza Z; Mukasheva, Togzhan D

    2015-06-01

    538 yeast strains were isolated from dark chestnut soil collected from under the plants of the legume family (Fabaceae). The greatest number of microorganisms is found at soil depth 10-20 cm. Among the 538 strains of yeast 77 (14.3%) strains demonstrated the ability to synthesize IAA. 15 strains were attributed to high IAA-producing yeasts (above 10 μg/ml). The most active strains were YA05 with 51.7 ± 2.1 μg/ml of IAA and YR07 with 45.3 ± 1.5 μg/ml. In the study of effect of incubation time on IAA production the maximum accumulation of IAA coincided with maximum rates of biomass: at 120 h for YR07 and at 144 h for strain YA05. IAA production increased when medium was supplemented with the L-tryptophan. 400 μg/ml of L-tryptophan showed maximum IAA production. 10 strains demonstrated the ability to inhibit the growth and development of phytopathogenic fungi. YA05 and YR07 strains formed the largest zones of inhibition compared to the other strains--from 21.6 ± 0.3 to 30.6 ± 0.5 mm. Maximum zone of inhibition was observed for YA05 against Phytophtora infestans and YR07 strains against Fusarium graminearum. YA05 and YR07 strains were identified as Aureobasidium pullulans YA05 (GenBank accession No JF160955) and Rhodotorula mucilaginosa YR07 (GenBank accession No JF160956). PMID:25843007

  14. Metabarcoding Analysis of Fungal Diversity in the Phyllosphere and Carposphere of Olive (Olea europaea).

    PubMed

    Abdelfattah, Ahmed; Li Destri Nicosia, Maria Giulia; Cacciola, Santa Olga; Droby, Samir; Schena, Leonardo

    2015-01-01

    The fungal diversity associated with leaves, flowers and fruits of olive (Olea europaea) was investigated in different phenological stages (May, June, October and December) using an implemented metabarcoding approach. It consisted of the 454 pyrosequencing of the fungal ITS2 region and the subsequent phylogenetic analysis of relevant genera along with validated reference sequences. Most sequences were identified up to the species level or were associated with a restricted number of related taxa enabling supported speculations regarding their biological role. Analyses revealed a rich fungal community with 195 different OTUs. Ascomycota was the dominating phyla representing 93.6% of the total number of detected sequences followed by unidentified fungi (3.6%) and Basidiomycota (2.8%). A higher level of diversity was revealed for leaves compared to flowers and fruits. Among plant pathogens the genus Colletotrichum represented by three species (C. godetiae syn. C. clavatum, C. acutatum s.s and C. karstii) was the most abundant on ripe fruits but it was also detected in other organs. Pseudocercospora cladosporioides was detected with a high frequency in all leaf samples and to a less extent in ripe fruits. A much lower relative frequency was revealed for Spilocaea oleagina and for other putative pathogens including Fusarium spp., Neofusicoccum spp., and Alternaria spp. Among non-pathogen taxa, Aureobasidium pullulans, the species complex of Cladosporium cladosporioides and Devriesia spp. were the most represented. This study highlights the existence of a complex fungal consortium including both phytopathogenic and potentially antagonistic microorganisms that can have a significant impact on olive productions. PMID:26132745

  15. Yeast identification in floral nectar of Mimulus aurantiacus (Invited)

    NASA Astrophysics Data System (ADS)

    Kyauk, C.; Belisle, M.; Fukami, T.

    2009-12-01

    Nectar is such a sugar-rich resource that serves as a natural habitat in which microbes thrive. As a result, yeasts arrive to nectar on the bodies of pollinators such as hummingbirds and bees. Yeasts use the sugar in nectar for their own needs when introduced. This research focuses on the identification of different types of yeast that are found in the nectar of Mimulus aurantiacus (commonly known as sticky monkey-flower). Unopened Mimulus aurantiacus flower buds were tagged at Jasper Ridge and bagged three days later. Floral nectar was then extracted and plated on potato dextrose agar. Colonies on the plates were isolated and DNA was extracted from each sample using QIAGEN DNeasy Plant Mini Kit. The DNA was amplified through PCR and ran through gel electrophoresis. The PCR product was used to clone the nectar samples into an E.coli vector. Finally, a phylogenetic tree was created by BLAST searching sequences in GenBank using the Internal Transcribed Space (ITS) locus. It was found that 18 of the 50 identified species were Candida magnifica, 14 was Candida rancensis, 6 were Crytococcus albidus and there were 3 or less of the following: Starmella bombicola, Candida floricola, Aureobasidium pullulans, Pichia kluyvera, Metschnikowa cibodaserisis, Rhodotorua colostri, and Malassezia globosa. The low diversity of the yeast could have been due to several factors: time of collection, demographics of Jasper Ridge, low variety of pollinators, and sugar concentration of the nectar. The results of this study serve as a necessary first step for a recently started research project on ecological interactions between plants, pollinators, and nectar-living yeast. More generally, this research studies the use of the nectar-living yeast community as a natural microcosm for addressing basic questions about the role of dispersal and competitive and facilitative interactions in ecological succession.

  16. Sequence-based Analysis of the Vitis vinifera L. cv Cabernet Sauvignon Grape Must Mycobiome in Three South African Vineyards Employing Distinct Agronomic Systems.

    PubMed

    Setati, Mathabatha E; Jacobson, Daniel; Bauer, Florian F

    2015-01-01

    Recent microbiomic research of agricultural habitats has highlighted tremendous microbial biodiversity associated with such ecosystems. Data generated in vineyards have furthermore highlighted significant regional differences in vineyard biodiversity, hinting at the possibility that such differences might be responsible for regional differences in wine style and character, a hypothesis referred to as "microbial terroir." The current study further contributes to this body of work by comparing the mycobiome associated with South African (SA) Cabernet Sauvignon grapes in three neighboring vineyards that employ different agronomic approaches, and comparing the outcome with similar data sets from Californian vineyards. The aim of this study was to fully characterize the mycobiomes associated with the grapes from these vineyards. The data revealed approximately 10 times more fungal diversity than what is typically retrieved from culture-based studies. The Biodynamic vineyard was found to harbor a more diverse fungal community (H = 2.6) than the conventional (H = 2.1) and integrated (H = 1.8) vineyards. The data show that ascomycota are the most abundant phylum in the three vineyards, with Aureobasidium pullulans and its close relative Kabatiella microsticta being the most dominant fungi. This is the first report to reveal a high incidence of K. microsticta in the grape/wine ecosystem. Different common wine yeast species, such as Metschnikowia pulcherrima and Starmerella bacillaris dominated the mycobiome in the three vineyards. The data show that the filamentous fungi are the most abundant community in grape must although they are not regarded as relevant during wine fermentation. Comparison of metagenomic datasets from the three SA vineyards and previously published data from Californian vineyards revealed only 25% of the fungi in the SA dataset was also present in the Californian dataset, with greater variation evident amongst ubiquitous epiphytic fungi. PMID:26648930

  17. The diversity and extracellular enzymatic activities of yeasts isolated from water tanks of Vriesea minarum, an endangered bromeliad species in Brazil, and the description of Occultifur brasiliensis f.a., sp. nov.

    PubMed

    Gomes, Fátima C O; Safar, Silvana V B; Marques, Andrea R; Medeiros, Adriana O; Santos, Ana Raquel O; Carvalho, Cláudia; Lachance, Marc-André; Sampaio, José Paulo; Rosa, Carlos A

    2015-02-01

    The diversity of yeast species collected from the bromeliad tanks of Vriesea minarum, an endangered bromeliad species, and their ability to produce extracellular enzymes were studied. Water samples were collected from 30 tanks of bromeliads living in a rupestrian field site located at Serrada Piedade, Minas Gerais state, Brazil, during both the dry and rainy seasons. Thirty-six species were isolated, representing 22 basidiomycetous and 14 ascomycetous species. Occultifur sp., Cryptococcus podzolicus and Cryptococcus sp. 1 were the prevalent basidiomycetous species. The yeast-like fungus from the order Myriangiales, Candida silvae and Aureobasidium pullulans were the most frequent ascomycetous species. The diversity of the yeast communities obtained between seasons was not significantly different, but the yeast composition per bromeliad was different between seasons. These results suggest that there is significant spatial heterogeneity in the composition of populations of the yeast communities within bromeliad tanks, independent of the season. Among the 352 yeast isolates tested, 282 showed at least one enzymatic activity. Protease activity was the most widely expressed extracellular enzymatic activity, followed by xylanase, amylase, pectinase and cellulase activities. These enzymes may increase the carbon and nitrogen availability for the microbial food web in the bromeliad tank of V. minarum. Sequence analyses revealed the existence of 10 new species, indicating that bromeliad tanks are important sources of new yeasts. The novel species Occultifur brasiliensis, f.a., sp. nov., is proposed to accommodate the most frequently isolated yeast associated with V. minarum. The type strain of O. brasiliensis, f.a., sp. nov. is UFMG-CM-Y375(T) (= CBS 12687(T)). The Mycobank number is MB 809816.

  18. Leukocyte mimetic polysaccharide microparticles tracked in vivo on activated endothelium and in abdominal aortic aneurysm.

    PubMed

    Bonnard, Thomas; Serfaty, Jean-Michel; Journé, Clément; Ho Tin Noe, Benoît; Arnaud, Denis; Louedec, Liliane; Derkaoui, Sidi Mohammed; Letourneur, Didier; Chauvierre, Cédric; Le Visage, Catherine

    2014-08-01

    We have developed injectable microparticles functionalized with fucoidan, in which sulfated groups mimic the anchor sites of P-selectin glycoprotein ligand-1 (PSGL-1), one of the principal receptors supporting leukocyte adhesion. These targeted microparticles were combined with a fluorescent dye and a T2(∗) magnetic resonance imaging (MRI) contrast agent, and then tracked in vivo with small animal imaging methods. Microparticles of 2.5μm were obtained by a water-in-oil emulsification combined with a cross-linking process of polysaccharide dextran, fluorescein isothiocyanate dextran, pullulan and fucoidan mixed with ultrasmall superparamagnetic particles of iron oxide. Fluorescent intravital microscopy observation revealed dynamic adsorption and a leukocyte-like behaviour of fucoidan-functionalized microparticles on a calcium ionophore induced an activated endothelial layer of a mouse mesentery vessel. We observed 20times more adherent microparticles on the activated endothelium area after the injection of functionalized microparticles compared to non-functionalized microparticles (197±11 vs. 10±2). This imaging tool was then applied to rats presenting an elastase perfusion model of abdominal aortic aneurysm (AAA) and 7.4T in vivo MRI was performed. Visual analysis of T2(∗)-weighted MR images showed a significant contrast enhancement on the inner wall of the aneurysm from 30min to 2h after the injection. Histological analysis of AAA cryosections revealed microparticles localized inside the aneurysm wall, in the same areas in which immunostaining shows P-selectin expression. The developed leukocyte mimetic imaging tool could therefore be relevant for molecular imaging of vascular diseases and for monitoring biologically active areas prone to rupture in AAA. PMID:24769117

  19. Development and characterization of pharmacokinetic parameters of fast-dissolving films containing levocetirizine.

    PubMed

    Choudhary, Dhagla R; Patel, Vishnu A; Chhalotiya, Usmangani K; Patel, Harsha V; Kundawala, Aliasgar J

    2012-09-01

    A fast-dissolving film containing levocetirizine, a non-sedative antihistamine drug, was developed using pullulan, xanthan gum, propylene glycol, and tween 80 as the base materials. The drug content of the prepared films was within an acceptable limit as prescribed by the USP. The film exhibited excellent stability for four months when stored at 40 °C and 75% humidity. In vitro dissolution studies suggested a rapid disintegration, in which most of levocetirizine (93.54 ± 3.9%) dissolved within 90 seconds after insertion into the medium. Subsequently, Sprague-Dawley rats were used to compare the pharmacokinetic properties of the film preparation administered to the oral cavity, to those with oral administration of the pure drug solution. The pharmacokinetic parameters were similar between the two groups in which AUC(0-t) (ng h/ml), AUC(0-∞) (ng h/ml) C(max) (ng/ml), T(max) (min), K(el) (h(-1)), and t(1/2) (h) of the reference were 452.033 ± 43.68, 465.78 ± 48.16, 237.16 ± 19.87, 30, 0.453 ± 0.051, and 1.536 ± 0.118, respectively, for the film formulation 447.233 ± 46.24, 458.22 ± 46.74, 233.32 ± 17.19, 30, 0.464 ± 0.060, and 1.496 ± 0.293, respectively. These results suggest that the present levocetirizine containing fast-dissolving film is likely to become one of the choices to treat different allergic conditions. PMID:23008821

  20. Abdominal Aortic Aneurysms Targeted by Functionalized Polysaccharide Microparticles: a new Tool for SPECT Imaging

    PubMed Central

    Bonnard, Thomas; Yang, Gonord; Petiet, Anne; Ollivier, Véronique; Haddad, Oualid; Arnaud, Denis; Louedec, Liliane; Bachelet-Violette, Laure; Derkaoui, Sidi Mohammed; Letourneur, Didier; Chauvierre, Cedric; Le Visage, Catherine

    2014-01-01

    Aneurysm diagnostic is nowadays limited by the lack of technology that enables early detection and rupture risk prediction. New non invasive tools for molecular imaging are still required. In the present study, we present an innovative SPECT diagnostic tool for abdominal aortic aneurysm (AAA) produced from injectable polysaccharide microparticles radiolabeled with technetium 99m (99mTc) and functionalized with fucoidan, a sulfated polysaccharide with the ability to target P-Selectin. P-Selectin is a cell adhesion molecule expressed on activated endothelial cells and platelets which can be found in the thrombus of aneurysms, as well as in other vascular pathologies. Microparticles with a maximum hydrodynamic diameter of 4 µm were obtained by crosslinking the polysaccharides dextran and pullulan. They were functionalized with fucoidan. In vitro interactions with human activated platelets were assessed by flow cytometry that demonstrated a specific affinity of fucoidan functionalized microparticles for P-Selectin expressed by activated platelets. For in vivo AAA imaging, microparticles were radiolabeled with 99mTc and intravenously injected into healthy and AAA rats obtained by elastase perfusion through the aorta wall. Animals were scanned by SPECT imaging. A strong contrast enhancement located in the abdominal aorta of AAA rats was obtained, while no signal was obtained in healthy rats or in AAA rats after injection of non-functionalized control microparticles. Histological studies revealed that functionalized radiolabeled polysaccharide microparticles were localized in the AAA wall, in the same location where P-Selectin was expressed. These microparticles therefore constitute a promising SPECT imaging tool for AAA and potentially for other vascular diseases characterized by P-Selectin expression. Future work will focus on validating the efficiency of the microparticles to diagnose these other pathologies and the different stages of AAA. Incorporation of a