Fast Pyrolysis Behavior of Banagrass as a Function of Temperature and Volatiles Residence Time in a Fluidized Bed Reactor

DOE PAGES

Morgan, Trevor James; Turn, Scott Q.; George, Anthe

2015-08-26

A reactor was designed and commissioned to study the fast pyrolysis behavior of banagrass as a function of temperature and volatiles residence time. Four temperatures between 400 and 600°C were examined as well as four residence times between ~1.0 and 10 seconds. Pyrolysis product distributions of bio-oil, char and permanent gases were determined at each reaction condition. The elemental composition of the bio-oils and chars was also assessed. The greatest bio-oil yield was recorded when working at 450°C with a volatiles residence time of 1.4 s, ~37 wt% relative to the dry ash free feedstock (excluding pyrolysis water). The amountsmore » of char (organic fraction) and permanent gases under these conditions are ~4 wt% and 8 wt% respectively. The bio-oil yield stated above is for 'dry' bio-oil after rotary evaporation to remove solvent, which results in volatiles and pyrolysis water being removed from the bio-oil. The material removed during drying accounts for the remainder of the pyrolysis products. The 'dry' bio-oil produced under these conditions contains ~56 wt% carbon which is ~40 wt% of the carbon present in the feedstock. The oxygen content of the 450°C, 1.4 s 'dry' bio-oil is ~38 wt%, which accounts for ~33 wt% of the oxygen in the feedstock. At higher temperature or longer residence time less bio-oil and char is recovered and more gas and light volatiles are produced. Increasing the temperature has a more significant effect on product yields and composition than increasing the volatiles residence time. At 600°C and a volatiles residence time of 1.2 seconds the bio-oil yield is ~21 wt% of the daf feedstock, with a carbon content of 64 wt% of the bio-oil. The bio-oil yield from banagrass is significantly lower than from woody biomass or grasses such as switchgrass or miscanthus, but is similar to barley straw. In conclusion, the reason for the low bio-oil yield from banagrass is thought to be related to its high ash content (8.5 wt% dry basis) and high concentration of alkali and alkali earth metals (totaling ~2.8 wt% relative to the dry feedstock) which are catalytic and increase cracking reactions during pyrolysis.« less

Fast Pyrolysis Behavior of Banagrass as a Function of Temperature and Volatiles Residence Time in a Fluidized Bed Reactor

PubMed Central

Morgan, Trevor James; Turn, Scott Q.; George, Anthe

2015-01-01

A reactor was designed and commissioned to study the fast pyrolysis behavior of banagrass as a function of temperature and volatiles residence time. Four temperatures between 400 and 600°C were examined as well as four residence times between ~1.0 and 10 seconds. Pyrolysis product distributions of bio-oil, char and permanent gases were determined at each reaction condition. The elemental composition of the bio-oils and chars was also assessed. The greatest bio-oil yield was recorded when working at 450°C with a volatiles residence time of 1.4 s, ~37 wt% relative to the dry ash free feedstock (excluding pyrolysis water). The amounts of char (organic fraction) and permanent gases under these conditions are ~4 wt% and 8 wt% respectively. The bio-oil yield stated above is for 'dry' bio-oil after rotary evaporation to remove solvent, which results in volatiles and pyrolysis water being removed from the bio-oil. The material removed during drying accounts for the remainder of the pyrolysis products. The 'dry' bio-oil produced under these conditions contains ~56 wt% carbon which is ~40 wt% of the carbon present in the feedstock. The oxygen content of the 450°C, 1.4 s 'dry' bio-oil is ~38 wt%, which accounts for ~33 wt% of the oxygen in the feedstock. At higher temperature or longer residence time less bio-oil and char is recovered and more gas and light volatiles are produced. Increasing the temperature has a more significant effect on product yields and composition than increasing the volatiles residence time. At 600°C and a volatiles residence time of 1.2 seconds the bio-oil yield is ~21 wt% of the daf feedstock, with a carbon content of 64 wt% of the bio-oil. The bio-oil yield from banagrass is significantly lower than from woody biomass or grasses such as switchgrass or miscanthus, but is similar to barley straw. The reason for the low bio-oil yield from banagrass is thought to be related to its high ash content (8.5 wt% dry basis) and high concentration of alkali and alkali earth metals (totaling ~2.8 wt% relative to the dry feedstock) which are catalytic and increase cracking reactions during pyrolysis. PMID:26308860

Fast Pyrolysis Behavior of Banagrass as a Function of Temperature and Volatiles Residence Time in a Fluidized Bed Reactor

DOE Office of Scientific and Technical Information (OSTI.GOV)

Morgan, Trevor James; Turn, Scott Q.; George, Anthe

A reactor was designed and commissioned to study the fast pyrolysis behavior of banagrass as a function of temperature and volatiles residence time. Four temperatures between 400 and 600°C were examined as well as four residence times between ~1.0 and 10 seconds. Pyrolysis product distributions of bio-oil, char and permanent gases were determined at each reaction condition. The elemental composition of the bio-oils and chars was also assessed. The greatest bio-oil yield was recorded when working at 450°C with a volatiles residence time of 1.4 s, ~37 wt% relative to the dry ash free feedstock (excluding pyrolysis water). The amountsmore » of char (organic fraction) and permanent gases under these conditions are ~4 wt% and 8 wt% respectively. The bio-oil yield stated above is for 'dry' bio-oil after rotary evaporation to remove solvent, which results in volatiles and pyrolysis water being removed from the bio-oil. The material removed during drying accounts for the remainder of the pyrolysis products. The 'dry' bio-oil produced under these conditions contains ~56 wt% carbon which is ~40 wt% of the carbon present in the feedstock. The oxygen content of the 450°C, 1.4 s 'dry' bio-oil is ~38 wt%, which accounts for ~33 wt% of the oxygen in the feedstock. At higher temperature or longer residence time less bio-oil and char is recovered and more gas and light volatiles are produced. Increasing the temperature has a more significant effect on product yields and composition than increasing the volatiles residence time. At 600°C and a volatiles residence time of 1.2 seconds the bio-oil yield is ~21 wt% of the daf feedstock, with a carbon content of 64 wt% of the bio-oil. The bio-oil yield from banagrass is significantly lower than from woody biomass or grasses such as switchgrass or miscanthus, but is similar to barley straw. In conclusion, the reason for the low bio-oil yield from banagrass is thought to be related to its high ash content (8.5 wt% dry basis) and high concentration of alkali and alkali earth metals (totaling ~2.8 wt% relative to the dry feedstock) which are catalytic and increase cracking reactions during pyrolysis.« less

Production of ajmalicine and ajmaline in hairy root cultures of Rauvolfia micrantha Hook f., a rare and endemic medicinal plant.

PubMed

Sudha, C G; Obul Reddy, B; Ravishankar, G A; Seeni, S

2003-04-01

Hairy roots of Rauvolfia micrantha were induced from hypocotyl explants of 2-3 weeks old aseptic seedlings using Agrobacterium rhizogenes ATCC 15834. Hairy roots grown in half-strength Murashige & Skoog (MS) medium with 0.2 mg indole 3-butyric acid l-1 and 0.1 mg alpha-naphthaleneacetic acid l-1 produced more ajmaline (0.01 mg g-1 dry wt) and ajmalicine (0.006 mg g-1 dry wt) than roots grown in auxin-free medium. Ajmaline (0.003 mg g-1 dry wt) and ajmalicine (0.0007 mg g-1 dry wt) were also produced in normal root cultures. This is the first report of production of ajmaline and ajmalicine in hairy root cultures of Rauvolfia micrantha.

A murine inhalation model to characterize pulmonary exposure to dry Aspergillus fumigatus conidia.

PubMed

Buskirk, Amanda D; Green, Brett J; Lemons, Angela R; Nayak, Ajay P; Goldsmith, W Travis; Kashon, Michael L; Anderson, Stacey E; Hettick, Justin M; Templeton, Steven P; Germolec, Dori R; Beezhold, Donald H

2014-01-01

Most murine models of fungal exposure are based on the delivery of uncharacterized extracts or liquid conidia suspensions using aspiration or intranasal approaches. Studies that model exposure to dry fungal aerosols using whole body inhalation have only recently been described. In this study, we aimed to characterize pulmonary immune responses following repeated inhalation of conidia utilizing an acoustical generator to deliver dry fungal aerosols to mice housed in a nose only exposure chamber. Immunocompetent female BALB/cJ mice were exposed to conidia derived from Aspergillus fumigatus wild-type (WT) or a melanin-deficient (Δalb1) strain. Conidia were aerosolized and delivered to mice at an estimated deposition dose of 1×105 twice a week for 4 weeks (8 total). Histopathological and immunological endpoints were assessed 4, 24, 48, and 72 hours after the final exposure. Histopathological analysis showed that conidia derived from both strains induced lung inflammation, especially at 24 and 48 hour time points. Immunological endpoints evaluated in bronchoalveolar lavage fluid (BALF) and the mediastinal lymph nodes showed that exposure to WT conidia led to elevated numbers of macrophages, granulocytes, and lymphocytes. Importantly, CD8+ IL17+ (Tc17) cells were significantly higher in BALF and positively correlated with germination of A. fumigatus WT spores. Germination was associated with specific IgG to intracellular proteins while Δalb1 spores elicited antibodies to cell wall hydrophobin. These data suggest that inhalation exposures may provide a more representative analysis of immune responses following exposures to environmentally and occupationally prevalent fungal contaminants.

Degradation of anionic surfactants during drying of UASBR sludges on sand drying beds.

PubMed

Mungray, Arvind Kumar; Kumar, Pradeep

2008-09-01

Anionic surfactant (AS) concentrations in wet up-flow anaerobic sludge blanket reactor (UASBR) sludges from five sewage treatment plants (STPs) were found to range from 4480 to 9,233 mg kg(-1)dry wt. (average 7,347 mg kg(-1)dry wt.) over a period of 18 months. After drying on sand drying beds (SDBs), AS in dried-stabilized sludges averaged 1,452 mg kg(-1)dry wt., a reduction of around 80%. The kinetics of drying followed simple first-order reduction of moisture with value of drying constant (k(d))=0.051 d(-1). Reduction of AS also followed first-order kinetics. AS degradation rate constant (k(AS)) was found to be 0.034 d(-1) and half-life of AS as 20 days. The order of rates of removal observed was k(d)>k(AS)>k(COD)>k(OM) (drying >AS degradation>COD reduction>organic matter reduction). For the three applications of dried-stabilized sludges (soil, agricultural soil, grassland), values of risk quotient (RQ) were found to be <1, indicating no risk.

Study of metals concentration levels in Patella piperata throughout the Canary Islands, Spain.

PubMed

Bergasa, Oscar; Ramírez, Rubén; Collado, Cayetano; Hernández-Brito, J Joaquín; Gelado-Caballero, María Dolores; Rodríguez-Somozas, María; Haroun, Ricardo J

2007-04-01

In order to assess the extent of metal contamination at rocky shores of the Canarian Archipelago, metal concentrations have been measured in Patella piperata (Gould, 1846), using the standard atomic absorption spectrophotometer technique. Ranges of elements concentrations measured (in microg g(-1)) found in the biota were: Cd (0.36 +/- 0.26 microg g(-1) dry wt.), Cu (2.05 +/- 0.91 dry wt.), Pb (1.57 +/- 1.14 microg g(-1)dry wt.) and Zn (10.37 +/- 4.60 microg g(-1) dry wt.). Variation in metal concentrations in Patella, was tested by using non-parametric statistical methods. Cd content had a maximum in the Archipelago Chinijo, northward of Lanzarote Island. The metal concentrations recorded at the clean stations may be considered carefully if they are used like background levels.

Electrospun oriented gelatin-hydroxyapatite fiber scaffolds for bone tissue engineering.

PubMed

Salifu, Ali A; Lekakou, Constantina; Labeed, Fatima H

2017-07-01

Tissue engineering of human fetal osteoblast cells was investigated on gelatin-hydroxyapatite (HA), crosslinked, electrospun oriented fiber scaffolds at the different HA concentrations of 0, 10, 20, and 25 wt % in the dry fibers and different fiber diameter, pore size and porosity of scaffolds. Rheological tests and proton nuclear magnetic resonance spectroscopy were conducted for all solutions used for electrospinning. It was found that 25 wt % HA-gelatin scaffolds electrospun at 20 kV led to the greatest cell attachment, cell proliferation and extracellular matrix (ECM) production while fiber orientation improved the mechanical properties, where crosslinked electrospun 25 wt % HA-gelatin fiber scaffolds yielded a Young's modulus in the range of 0.5-0.9 GPa and a tensile strength in the range of 4-10 MPa in the fiber direction for an applied voltage of 20-30 kV, respectively, in the electrospinning of scaffolds. Biological characterization of cell seeded scaffolds yielded the rate of cell growth and ECM (collagen and calcium) production by the cells as a function of time; it included cell seeding efficiency tests, alamar blue cell proliferation assay, alkaline phosphate (ALP) assay, collagen assay, calcium colorimetric assay, fluorescence microscopy for live and dead cells, and scanning electron microscopy for cell culture from 1 to 18 days. After 18 days, cells seeded and grown on the 25 wt % HA-gelatin scaffold, electrospun at 20 kV, reached production of collagen at 370 μg/L and calcium production at 0.8 mM. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1911-1926, 2017. © 2017 Wiley Periodicals, Inc.

Interleukin-12- and interferon-gamma-mediated natural killer cell activation by Agaricus blazei Murill.

PubMed

Yuminamochi, Eri; Koike, Taisuke; Takeda, Kazuyoshi; Horiuchi, Isao; Okumura, Ko

2007-06-01

Dried fruiting bodies of Agaricus blazei Murill (A. blazei) and its extracts have generally used as complementary and alternative medicines (CAMs). Here, we report that the oral administration of A. blazei augmented cytotoxicity of natural killer (NK) cells in wild-type (WT) C57BL/6, C3H/HeJ, and BALB/c mice. Augmented cytotoxicity was demonstrated by purified NK cells from treated wild-type (WT) and RAG-2-deficient mice, but not from interferon-gamma (IFN-gamma) deficient mice. NK cell activation and IFN-gamma production was also observed in vitro when dendritic cell (DC)-rich splenocytes of WT mice were coincubation with an extract of A. blazei. Both parameters were largely inhibited by neutralizing anti-interleukin-12 (IL-12) monoclonal antibody (mAb) and completely inhibited when anti-IL-12 mAb and anti-IL-18 mAb were used in combination. An aqueous extract of the hemicellulase-digested compound of A. blazei particle; (ABPC) induced IFN-gamma production more effectively, and this was completely inhibited by anti-IL-12 mAb alone. NK cell cytotoxicty was augmented with the same extracts, again in an IL-12 and IFN-gamma-dependent manner. These results clearly demonstrated that A. blazei and ABPC augmented NK cell activation through IL-12-mediated IFN-gamma production.

Exposure to PCBs, through inhalation, dermal contact and dust ingestion at Taizhou, China--a major site for recycling transformers.

PubMed

Xing, Guan Hua; Liang, Ying; Chen, Ling Xuan; Wu, Sheng Chun; Wong, Ming Hung

2011-04-01

Air samples containing gaseous and particulate phases were collected from e-waste workplaces and residential areas of an intensive e-waste recycling area and compared with a reference site. The highest total concentration of PCBs was detected at transformer recycling workshops (17.6 ng m(-3)), followed by the residential area (3.37 ng m(-3)) at Taizhou, and the lowest was obtained at the residential area of the reference site, Lin'an (0.46 ng m(-3)). The same trend was also observed with regards to PCB levels in dust samples. The highest average PCBs level of 2824 ng g(-1) (dry wt) was found in the transformer recycling workshops, and was significantly higher than that of residential areas of Taizhou (572 ng g(-1) dry wt) and Lin'an (42.4 ng g(-1) dry wt). WHO-PCB-TEQ level in the workshops of Taizhou was 2216 pg TEQ(1998)g(-1) dry wt or 2159 pg TEQ(2005)g(-1) dry wt, due to the high abundance of PCB 126 (21.5 ng g(-1) dry wt), which contributed 97% or 99% of WHO-PCB-TEQs. The estimated intake of PCBs via dust ingestion and dermal absorption by transformer recycling workers were 77.5×10(-5) and 36.0×10(-5) pg WHO-PCB-TEQ(1998)kg(-1)d(-1), and 67.3×10(-5) and 31.3×10(-5) pg WHO-PCB-TEQ(2005)kg(-1)d(-1), respectively. Copyright © 2011. Published by Elsevier Ltd.

Hyperthyroidism results in increased glycolytic capacity in the rat heart. A 31P-NMR study.

PubMed

Seymour, A M; Eldar, H; Radda, G K

1990-11-12

We have investigated the metabolic adaptations that occur in the thyroxine-treated rat heart. Rats were made hyperthyroid by daily intra-peritoneal injections of thyroxine (35 micrograms/100 g body weight) over seven days. 31P-NMR investigations of isolated glucose-perfused isometric hearts showed that thyroxine treatment caused an increase in Pi (from 4.9 mumols.(g dry wt.)-1 in control hearts to 11.7 mumols.(g dry wt.)-1 in hyperthyroid hearts), a decrease in phosphocreatine (from 36.5 mumols.(g dry wt.)-1 to 21.8 mumols.(g dry wt.)-1) with no change in ATP or ADP concentrations under the same conditions of cardiac work. The unidirectional exchange flux Pi----ATP was measured by saturation transfer NMR in hyperthyroid rat hearts. This exchange (which has been shown to contain a significant glycolytic component) increased by 2.2-fold in thyroxine-treated hearts in comparison to control hearts (to 3.6 mumols.(g dry wt.)-1.s-1, from 1.6 mumols.(g dry wt.)-1.s-1). In parallel experiments, NMR analysis of extracts from hyperthyroid rat hearts showed significantly elevated levels of glucose 6-phosphate, and fructose 6-phosphate. Measurements of enzyme activities isolated from hyperthyroid and control tissue showed a 40% increase in phosphofructokinase activity. These data together with the increased concentration of Pi show that both glycolytic and glycogenolytic fluxes are increased in the hyperthyroid rat heart. This metabolic adaptation may be necessary to cope with the increased number and activity of Na+/K(+)-ATPase pumps that occur in response to thyroxine treatment.

Peroxiredoxin 6 gene-targeted mice show increased lung injury with paraquat-induced oxidative stress.

PubMed

Wang, Yan; Feinstein, Sheldon I; Manevich, Yefim; Ho, Ye-Shih; Fisher, Aron B

2006-01-01

Mice with knock-out of peroxiredoxin 6 (Prdx6), a recently described antioxidant enzyme, were evaluated for susceptibility to lung injury with paraquat (PQ) administration. With high dose PQ (30 mg/kg i.p.), all Prdx6-/- mice died (LT50 54 +/- 2.05 h, mean +/- SE) by 4 days, whereas 86% of the wild-type (WT) mice (C57BL/6) survived (n = 14). At 2 days after PQ, lung wet/dry weight ratio increased significantly (p < 0.05) to 7.57 +/- 0.37 in Prdx6-/- mice vs. 5.42 +/- 0.25 in WT mice. Total protein and nucleated cells in bronchoalveolar lavage fluid and TBARS and protein carbonyls in lung homogenate also showed more marked increases in Prdx6-/- mice. At 2.5 days after PQ, light microscopy of WT lungs showed mild injury while Prdx6-/- lungs showed epithelial cell necrosis, perivascular edema, and inflammatory cells. With low dose PQ (12.5 mg/kg), mortality and lung injury were less marked but were significantly greater with Prdx6-/- compared to WT mice. These results show that Prdx6-/- mice have increased susceptibility to lung injury with PQ administration. Thus, Prdx6 protects lungs against PQ toxicity as shown previously for hyperoxia, indicating that it functions as an important lung antioxidant enzyme.

The effect of rework content addition on the microstructure and viscoelastic properties of processed cheese.

PubMed

Černíková, Michaela; Nebesářová, Jana; Salek, Richardos Nikolaos; Popková, Romana; Buňka, František

2018-04-01

The aim of this work was to add various amounts of rework (0.0 to 20.0% wt/wt) to processed cheeses with a dry matter content of 36% (wt/wt) and fat with a dry matter content of 45% (wt/wt). The effect of the rework addition on the viscoelastic properties and microstructure of the processed cheeses was observed. The addition of rework (in this case, to processed cheese with a spreadable consistency) in the amounts of 2.5, 5.0, and 10.0% (wt/wt) increased the firmness of the processed cheese. With the further addition of rework, the consistency of the processed cheeses no longer differed significantly. The conclusions obtained by the measurement of viscoelastic properties were supported by cryo-scanning electron microscopy, where fat droplets in samples with added rework of over 10.0% (wt/wt) were smaller than fat droplets in processed cheeses with lower additions of rework. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Mercury correlations among six tissues for four waterbird species breeding in San Francisco Bay, California, USA

USGS Publications Warehouse

Eagles-Smith, Collin A.; Ackerman, Joshua T.; Adelsbach, T.L.; Takekawa, John Y.; Miles, A.K.; Keister, R.A.

2008-01-01

Despite a large body of research concerning mercury (Hg) in birds, no single tissue has been used consistently to assess Hg exposure, and this has hampered comparisons across studies. We evaluated the relationships of Hg concentrations among tissues in four species of waterbirds (American avocets [Recurvirostra americana], black-necked stilts [Himantopus mexicanus], Caspian terns [Hydroprogne caspia; formerly Sterna caspia], and Forster's terns [Sterna forsteri]) and across three life stages (prebreeding adults, breeding adults, and chicks) in San Francisco Bay, California, USA. Across species and life stages, Hg concentrations (least square mean ?? standard error) were highest in head feathers (6.45 ?? 0.31 ??g/g dry wt) and breast feathers (5.76 ?? 0.28 ??g/g dry wt), followed by kidney (4.54 ?? 0.22 ??g/g dry wt), liver (4.43 ?? 0.21 ??g/g dry wt), blood (3.10 ?? 0.15 ??g/g dry wt), and muscle (1.67 ?? 0.08 ??g/g dry wt). Relative Hg distribution among tissues, however, differed by species and life stage. Mercury concentrations were highly correlated among internal tissues (r 2 ??? 0.89). Conversely, the relationships between Hg in feathers and internal tissues were substantially weaker (r2 ??? 0.42). Regression slopes sometimes differed among species and life stages, indicating that care must be used when predicting Hg concentrations in one tissue based on those in another. However, we found good agreement between predictions made using a general tissue-prediction equation and more specific equations developed for each species and life stage. Finally, our results suggest that blood is an excellent, nonlethal predictor of Hg concentrations in internal tissues but that feathers are relatively poor indicators of Hg concentrations in internal tissues. ?? 2008 SETAC Printed in the USA.

Simulation of co-incineration of sewage sludge with municipal solid waste in a grate furnace incinerator.

PubMed

Lin, Hai; Ma, Xiaoqian

2012-03-01

Incineration is one of the most important methods in the resource recovery disposal of sewage sludge. The combustion characteristics of sewage sludge and an increasing number of municipal solid waste (MSW) incineration plants provide the possibility of co-incineration of sludge with MSW. Computational fluid dynamics (CFD) analysis was used to verify the feasibility of co-incineration of sludge with MSW, and predict the effect of co-incineration. In this study, wet sludge and semi-dried sludge were separately blended with MSW as mixed fuels, which were at a co-incineration ratios of 5 wt.% (wet basis, the same below), 10 wt.%, 15 wt.%, 20 wt.% and 25 wt.%. The result indicates that co-incineration of 10 wt.% wet sludge with MSW can ensure the furnace temperature, the residence time and other vital items in allowable level, while 20 wt.% of semi-dried sludge can reach the same standards. With lower moisture content and higher low heating value (LHV), semi-dried sludge can be more appropriate in co-incineration with MSW in a grate furnace incinerator. Copyright © 2011 Elsevier Ltd. All rights reserved.

The Production of Biodiesel from Cottonseed Oil Using Rhizopus oryzae Whole Cell Biocatalysts

NASA Astrophysics Data System (ADS)

Athalye, Sneha Kishor

Biodiesel is an environmentally friendly alternative to fossil fuels which have become increasingly expensive in recent times. An alternate approach to alkaline biodiesel production is needed as catalyst miscibility with the glycerol by-product, generation of large amounts of waste water, and saponification of the feedstock are major disadvantages associated with the process. Lipases are water soluble enzymes which act as catalysts in many lipid based reactions. Reuse of lipases can significantly reduce cost of enzymatic biodiesel production; however retention of lipolytic activity still remains a challenge. Use of microbial cells immobilized on various surfaces like sponge, foam and plastics as biocatalysts instead of extracted enzyme could help overcome this problem. A novel, rigid biomass support with high surface area made from recyclable polyethylene (Bioblok(TM)) was used in this study. Several fungal and bacterial species have been reported to possess appreciable levels of lipase activity. The biomass production and immobilization as well as lipase activity of three different species; Candida rugosa (ATCC #38772), Aspergillus oryzae (ATCC #58299), and Rhizopus oryzae (ATTC #34612) were tested. C. rugosa did not attach well to the support particles while A.oryzae had lower biomass accumulation of 6.1 g (dry cell wt)/L compared to 11.8 g (dry cell wt)/L for R.oryzae. Hence Rhizopus oryzae, fungal specie with cell surface bound lipase was selected for the current study. The study investigated the influence of media composition and growth time of the R.oryzae whole cell biocatalysts, immobilized on the BSPs, for FAME production from cottonseed oil. R.oryzae BSPs grown in basal media supplemented with 1% (w/v) of glucose or oil or both for 48 h, 72 h or 90 h were used in a 36 h transesterification reaction with cottonseed oil and methanol. BSPs grown in both glucose and oil supplemented medium for 72 h had the highest conversion of 22.4% (wt/wt) and a biomass accumulation of 15.6 g (dry cell wt)/L. A reduction in dynamic viscosity of the reaction mixture from 47.3 centipoise to 30.6 centipoise was observed. The impact of moisture addition to the reaction mixture and use of ethanol as acylating agent on R.oryzae BSP fatty acid alkyl ester production was also tested. The presence of 10 wt % moisture in the reaction system had a significant effect (p ≤ 0.05) on the transesterification reaction with ethanol unlike methanol. Fatty acid ethyl ester concentration tripled from 39.3 to 129.1 g/L when moisture was added during transesterification .When oil to acyl acceptor ratio was increased from 1:3 and 1:6 to determine effect of excess alcohol on conversion, an ester conversion of 128.1 g/L for methanol and 129.1 g/L for ethanol were observed. Use of excess amount of acylating agent had a significant adverse effect (p ≤ 0.05) on the overall FAAE production due to deactivation of lipases on contact with large amounts of insoluble alcohol in the oil phase of the reaction. The effect of short chain alcohols on the enzymatic transesterification of cottonseed oil using freeze dried Rhizopus oryzae biomass was examined with and without water addition using methanol, ethanol, 1-Propanol and 1-Butanol at various molar ratios. 1- Butanol in the absence of water resulted in a significantly higher (p . 0.1) conversion of cottonseed oil to 12.5 % fatty acid butyl esters (FABEs). Addition of 10 % water to the reaction mixture significantly reduced (p ≤ 0.1) conversion. No significant difference (p > 0.1) between the conversions was observed for time points after 24 h for a 72 h reaction. 1- Butanol in ratios higher than 3:1 to cottonseed oil had a significant impact (p ≤ 0.1) on conversion. Increasing the amount of biomass used during the reaction lead to significantly higher conversion (p ≤ 0.1). The highest conversion of 27.9 % was observed for the transesterification reaction between cottonseed oil and 1-Butanol, in a 1:6 molar ratio, in the absence of water, when 100 mg of freeze dried R.oryzae biomass was used. The present study concluded that R.oryzae biomass attached better to the Bioblok. material and had highest lipase activity when grown in presence of oil and glucose for 72h. Addition of water to the reaction system gave higher fatty acid alkyl ester conversion when whole immobilized R.oryzae BSPs were used but absence of water promoted conversion with freeze dried biomass.

Comparison of organochlorine pesticides and polychlorinated biphenyls residues in vegetables, grain and soil from organic and conventional farming in Poland.

PubMed

Witczak, Agata; Abdel-Gawad, Hassan

2012-01-01

Organic and conventional crops were studied by identifying the relationship between persistent organic pollutants in cereals, vegetables and soil. The residues of organochlorine pesticides and polychlorinated biphenyls (PCBs) were determined in grains (rye and wheat), vegetables (carrots and beets) and soil collected from the fields. PCB residues recorded in the beets from organic farming were as high as 3.71 ppb dry weight (dry wt.), while in the soil from conventional farming of beets 0.53 ppb dry wt. Among vegetables, higher concentrations of pesticides were detected in organically grown beets (190.63 ppb dry wt.). Soil samples from the organic farming contained lower levels of organochlorine pesticide residues compared to the conventional farming. Taking into account toxicity equivalent (TEQ), the conventionally grown carrots accumulated the most toxic PCBs. Non-ortho and mono-ortho PCBs were also noted in the grain of conventionally grown rye and amounted to 3.05 pg-TEQ/g wet wt.

Interleukin-12- and interferon-γ-mediated natural killer cell activation by Agaricus blazei Murill

PubMed Central

Yuminamochi, Eri; Koike, Taisuke; Takeda, Kazuyoshi; Horiuchi, Isao; Okumura, Ko

2007-01-01

Dried fruiting bodies of Agaricus blazei Murill (A. blazei) and its extracts have generally used as complementary and alternative medicines (CAMs). Here, we report that the oral administration of A. blazei augmented cytotoxicity of natural killer (NK) cells in wild-type (WT) C57BL/6, C3H/HeJ, and BALB/c mice. Augmented cytotoxicity was demonstrated by purified NK cells from treated wild-type (WT) and RAG-2-deficient mice, but not from interferon-γ (IFN-γ) deficient mice. NK cell activation and IFN-γ production was also observed in vitro when dendritic cell (DC)-rich splenocytes of WT mice were coincubation with an extract of A. blazei. Both parameters were largely inhibited by neutralizing anti-interleukin-12 (IL-12) monoclonal antibody (mAb) and completely inhibited when anti-IL-12 mAb and anti-IL-18 mAb were used in combination. An aqueous extract of the hemicellulase-digested compound of A. blazei particle; (ABPC) induced IFN-γ production more effectively, and this was completely inhibited by anti-IL-12 mAb alone. NK cell cytotoxicty was augmented with the same extracts, again in an IL-12 and IFN-γ-dependent manner. These results clearly demonstrated that A. blazei and ABPC augmented NK cell activation through IL-12-mediated IFN-γ production. PMID:17346284

Application of Freeze-Dried Powders of Genetically Engineered Microbial Strains as Adsorbents for Rare Earth Metal Ions.

PubMed

Moriwaki, Hiroshi; Masuda, Reiko; Yamazaki, Yuki; Horiuchi, Kaoru; Miyashita, Mari; Kasahara, Jun; Tanaka, Tatsuhito; Yamamoto, Hiroki

2016-10-12

The adsorption behaviors of the rare earth metal ions onto freeze-dried powders of genetically engineered microbial strains were compared. Cell powders obtained from four kinds of strains, Bacillus subtilis 168 wild type (WT), lipoteichoic acid-defective (ΔLTA), wall teichoic acid-defective (ΔWTA), and cell wall hydrolases-defective (EFKYOJLp) strains, were used as an adsorbent of the rare earth metal ions at pH 3. The adsorption ability of the rare earth metal ions was in the order of EFKYOJLp > WT > ΔLTA > ΔWTA. The order was the same as the order of the phosphorus quantity of the strains. This result indicates that the main adsorption sites for the ions are the phosphate groups and the teichoic acids, LTA and WTA, that contribute to the adsorption of the rare earth metal ions onto the cell walls. The contribution of WTA was clearly greater than that of LTA. Each microbial powder was added to a solution containing 16 kinds of rare earth metal ions, and the removals (%) of each rare earth metal ion were obtained. The scandium ion showed the highest removal (%), while that of the lanthanum ion was the lowest for all the microbial powders. Differences in the distribution coefficients between the kinds of lanthanide ions by the EFKYOJLp and ΔWTA powders were greater than those of the other strains. Therefore, the EFKYOJLp and ΔWTA powders could be applicable for the selective extraction of the lanthanide ions. The ΔLTA powder coagulated by mixing with a rare earth metal ion, although no sedimentation of the WT or ΔWTA powder with a rare earth metal ion was observed under the same conditions. The EFKYOJLp powder was also coagulated, but its flocculating activity was lower than that of ΔLTA. The ΔLTA and EFKYOJLp powders have a long shape compared to those of the WT or ΔWTA strain. The shapes of the cells will play an important role in the sedimentation of the microbial powders with rare earth metal ions. As the results, three kinds of the genetically engineered microbial powders revealed unique adsorption behaviors of the rare earth metal ions.

Enumeration of viruses and prokaryotes in deep-sea sediments and cold seeps of the Gulf of Mexico

USGS Publications Warehouse

Kellogg, Christina A.

2010-01-01

Little is known about the distribution and abundance of viruses in deep-sea cold-seep environments. Like hydrothermal vents, seeps support communities of macrofauna that are sustained by chemosynthetic bacteria. Sediments close to these communities are hypothesized to be more microbiologically active and therefore to host higher numbers of viruses than non-seep areas. Push cores were taken at five types of Gulf of Mexico habitats at water depths below 1000 m using a remotely operated vehicle (ROV). The habitats included non-seep reference sediment, brine seeps, a microbial mat, an urchin field, and a pogonophoran worm community. Samples were processed immediately for enumeration of viruses and prokaryotes without the addition of a preservative. Prokaryote counts were an order of magnitude lower in sediments directly in contact with macrofauna (urchins, pogonophorans) compared to all other samples (107 vs. 108 cells g-1 dry weight) and were highest in areas of elevated salinity (brine seeps). Viral-Like Particle (VLP) counts were lowest in the reference sediments and pogonophoran cores (108 VLP g-1 dry wt), higher in brine seeps (109 VLP g-1 dry wt), and highest in the microbial mats (1010 VLP g-1 dry wt). Virus-prokaryote ratios (VPR) ranged from <5 in the reference sediment to >30 in the microbial mats and >60 in the urchin field. VLP counts and VPR were all significantly greater than those reported from sediments in the deep Mediterranean Sea and in most cases were higher than recent data from a cold-seep site near Japan. The high VPR suggest that greater microbial activity in or near cold-seep environments results in greater viral production and therefore higher numbers of viruses.

Enumeration of viruses and prokaryotes in deep-sea sediments and cold seeps of the Gulf of Mexico

NASA Astrophysics Data System (ADS)

Kellogg, Christina A.

2010-11-01

Little is known about the distribution and abundance of viruses in deep-sea cold-seep environments. Like hydrothermal vents, seeps support communities of macrofauna that are sustained by chemosynthetic bacteria. Sediments close to these communities are hypothesized to be more microbiologically active and therefore to host higher numbers of viruses than non-seep areas. Push cores were taken at five types of Gulf of Mexico habitats at water depths below 1000 m using a remotely operated vehicle (ROV). The habitats included non-seep reference sediment, brine seeps, a microbial mat, an urchin field, and a pogonophoran worm community. Samples were processed immediately for enumeration of viruses and prokaryotes without the addition of a preservative. Prokaryote counts were an order of magnitude lower in sediments directly in contact with macrofauna (urchins, pogonophorans) compared to all other samples (107 vs. 108 cells g-1 dry weight) and were highest in areas of elevated salinity (brine seeps). Viral-Like Particle (VLP) counts were lowest in the reference sediments and pogonophoran cores (108 VLP g-1 dry wt), higher in brine seeps (109 VLP g-1 dry wt), and highest in the microbial mats (1010 VLP g-1 dry wt). Virus-prokaryote ratios (VPR) ranged from <5 in the reference sediment to >30 in the microbial mats and >60 in the urchin field. VLP counts and VPR were all significantly greater than those reported from sediments in the deep Mediterranean Sea and in most cases were higher than recent data from a cold-seep site near Japan. The high VPR suggest that greater microbial activity in or near cold-seep environments results in greater viral production and therefore higher numbers of viruses.

Mechanical Properties of Calcium Fluoride-Based Composite Materials

PubMed Central

Kleczewska, Joanna; Pryliński, Mariusz; Podlewska, Magdalena; Sokołowski, Jerzy; Łapińska, Barbara

2016-01-01

Aim of the study was to evaluate mechanical properties of light-curing composite materials modified with the addition of calcium fluoride. The study used one experimental light-curing composite material (ECM) and one commercially available flowable light-curing composite material (FA) that were modified with 0.5–5.0 wt% anhydrous calcium fluoride. Morphology of the samples and uniformity of CaF2 distribution were analyzed using Scanning Electron Microscopy (SEM) and Energy Dispersive Spectroscopy (EDS). Mechanical properties were tested after 24-hour storage of specimens in dry or wet conditions. Stored dry ECM enriched with 0.5–1.0 wt% CaF2 showed higher tensile strength values, while water storage of all modified ECM specimens decreased their tensile strength. The highest Vickers hardness tested after dry storage was observed for 2.5 wt% CaF2 content in ECM. The addition of 2.0–5.0 wt% CaF2 to FA caused significant decrease in tensile strength after dry storage and overall tensile strength decrease of modified FA specimens after water storage. The content of 2.0 wt% CaF2 in FA resulted in the highest Vickers hardness tested after wet storage. Commercially available composite material (FA), unmodified with fluoride addition, demonstrated overall significantly higher mechanical properties. PMID:28004001

Water hyacinths for water quality improvement and biomass production

DOE Office of Scientific and Technical Information (OSTI.GOV)

Reddy, K.R.; Sutton, D.L.

The potential use of water hyacinth (Eichhornia crassipes (Mart.) Solms) for biomass production and for nutrient removal from waste waters is discussed. Warm climates in tropical and subtropical areas are conducive for establishing waste water treatment systems and biomass production farms with water hyacinth. Sources of nutrients available to culture water hyacinths include sewage effluent, agricultural drainage water, runoff from animal waste operations, methane digestor effluent, and water from eutrophic lakes and rivers. Growth rates of water hyacinths were found to be influenced by the nutrient composition of the water, plant density, solar radiation, and temperature. Annual yields of watermore » hyacinth biomass were found to range from 47 to 106 Mg dry wt ha/sup -1/ y/sup -1/, with approximately 50% of the biomass produced during May through August. A pond with a surface area of 2.65 ha and 1.0 m in depth containing 15 to 30 kg wet wt of water hyacinth per square meter (750-1500 g dry wt m/sup -2/) and a detention period of 7 d is adequate to treat 3785 m/sup 3/ d/sup -1/ (million gallons per day (mdg)) of sewage effluent. This hypothetical system would achieve 70 to 80% N removal, 40 to 50% P removal, and would produce a biomass yield of 690 to 1060 kg dry wt d/sup -1/ (13-20 g dry wt m/sup -2/ d/sup -1/). The biomass upon anaerobic digestion would yield 180 to 280 m/sup 3/ of methane/d. This is equivalent to 549 to 843 GJ ha/sup -1/ of energy from water hyacinth produced at a rate of 48 to 73 Mg dry wt ha/sup -1/ y/sup -1/.« less

Influence of a chlor-alkali superfund site on mercury bioaccumulation in periphyton and low-trophic level fauna

USGS Publications Warehouse

Buckman, Kate L.; Marvin-DiPasquale, Mark C.; Taylor, Vivien F.; Chalmers, Ann T.; Broadley, Hannah J.; Agee, Jennifer L.; Jackson, Brian P.; Chen, Celia Y.

2015-01-01

In Berlin, New Hampshire, USA, the Androscoggin River flows adjacent to a former chlor-alkali facility that is a US Environmental Protection Agency Superfund site and source of mercury (Hg) to the river. The present study was conducted to determine the fate and bioaccumulation of methylmercury (MeHg) to lower trophic-level taxa in the river. Surface sediment directly adjacent to the source showed significantly elevated MeHg (10–40× increase, mean ± standard deviation [SD]: 20.1 ± 24.8 ng g–1 dry wt) and total mercury (THg; 10–30× increase, mean ± SD: 2045 ± 2669 ng g–1 dry wt) compared with all other reaches, with sediment THg and MeHg from downstream reaches elevated (3–7× on average) relative to the reference (THg mean ± SD: 33.5 ± 9.33 ng g–1 dry wt; MeHg mean ± SD: 0.52 ± 0.21 ng g–1 dry wt). Water column THg concentrations adjacent to the point source for both particulate (0.23 ng L–1) and dissolved (0.76 ng L–1) fractions were 5-fold higher than at the reference sites, and 2-fold to 5-fold higher than downstream. Methylmercury production potential of periphyton material was highest (2–9 ng g–1 d–1 dry wt) adjacent to the Superfund site; other reaches were close to or below reporting limits (0. 1 ng g–1 d–1 dry wt). Total Hg and MeHg bioaccumulation in fauna was variable across sites and taxa, with no clear spatial patterns downstream of the contamination source. Crayfish, mayflies, and shiners showed a weak positive relationship with porewater MeHg concentration.

Influence of a chlor-alkali superfund site on mercury bioaccumulation in periphyton and low-trophic level fauna.

PubMed

Buckman, Kate L; Marvin-DiPasquale, Mark; Taylor, Vivien F; Chalmers, Ann; Broadley, Hannah J; Agee, Jennifer; Jackson, Brian P; Chen, Celia Y

2015-07-01

In Berlin, New Hampshire, USA, the Androscoggin River flows adjacent to a former chlor-alkali facility that is a US Environmental Protection Agency Superfund site and source of mercury (Hg) to the river. The present study was conducted to determine the fate and bioaccumulation of methylmercury (MeHg) to lower trophic-level taxa in the river. Surface sediment directly adjacent to the source showed significantly elevated MeHg (10-40× increase, mean ± standard deviation [SD]: 20.1 ± 24.8 ng g(-1) dry wt) and total mercury (THg; 10-30× increase, mean ± SD: 2045 ± 2669 ng g(-1) dry wt) compared with all other reaches, with sediment THg and MeHg from downstream reaches elevated (3-7× on average) relative to the reference (THg mean ± SD: 33.5 ± 9.33 ng g(-1) dry wt; MeHg mean ± SD: 0.52 ± 0.21 ng g(-1) dry wt). Water column THg concentrations adjacent to the point source for both particulate (0.23 ng L(-1)) and dissolved (0.76 ng L(-1)) fractions were 5-fold higher than at the reference sites, and 2-fold to 5-fold higher than downstream. Methylmercury production potential of periphyton material was highest (2-9 ng g(-1) d(-1) dry wt) adjacent to the Superfund site; other reaches were close to or below reporting limits (0. 1 ng g(-1) d(-1) dry wt). Total Hg and MeHg bioaccumulation in fauna was variable across sites and taxa, with no clear spatial patterns downstream of the contamination source. Crayfish, mayflies, and shiners showed a weak positive relationship with porewater MeHg concentration. © 2015 SETAC.

Value-added oil and animal feed production from corn-ethanol stillage using the oleaginous fungus Mucor circinelloides.

PubMed

Mitra, Debjani; Rasmussen, Mary L; Chand, Priyanka; Chintareddy, Venkat Reddy; Yao, Linxing; Grewell, David; Verkade, John G; Wang, Tong; van Leeuwen, J Hans

2012-03-01

This study highlights the potential of oleaginous fungus, Mucor circinelloides in adsorbing/assimilating oil and nutrients in thin stillage (TS), and producing lipid and protein-rich fungal biomass. Fungal cultivation on TS for 2 days in a 6-L airlift bioreactor, resulted in a 92% increase in oil yield from TS, and 20 g/L of fungal biomass (dry) with a lipid content of 46% (g of oil per 100g dry biomass). Reduction in suspended solids and soluble chemical oxygen demand (SCOD) in TS were 95% and 89%, respectively. The polyunsaturated fatty acids in fungal oil were 52% of total lipids. Fungal cells grown on Yeast Malt (YM) broth had a higher concentration of γ-linolenic acid (17 wt.%) than those grown on TS (1.4 wt.%). Supplementing TS with crude glycerol (10%, v/v) during the stationary growth phase led to a further 32% increase (from 46% to 61%) in cellular oil content. Copyright © 2011 Elsevier Ltd. All rights reserved.

Comparative study of wet and dry torrefaction of corn stalk and the effect on biomass pyrolysis polygeneration.

PubMed

Wang, Xianhua; Wu, Jing; Chen, Yingquan; Pattiya, Adisak; Yang, Haiping; Chen, Hanping

2018-06-01

Wet torrefaction (WT) possesses some advantages over dry torrefaction (DT). In this study, a comparative analysis of torrefied corn stalk from WT and DT was conducted along with an investigation of their pyrolysis properties under optimal conditions for biomass pyrolysis polygeneration. Compared with DT, WT removed 98% of the ash and retained twice the amount of hydrogen. The impacts of DT and WT on the biomass macromolecular structure was also found to be different using two-dimensional perturbation correlation infrared spectroscopy (2D-PCIS). WT preserved the active hydroxyl groups and rearranged the macromolecule structure to allow cellulose to be more ordered, while DT removed these active hydroxyl groups and formed inter-crosslinking structures in macromolecules. Correspondingly, the bio-char yield after WT was lower than DT but the bio-char quality was upgraded due to high ash removal. Furthermore, higher bio-oil yield, higher sugar content, and higher H 2 generation, were obtained after WT. Copyright © 2018 Elsevier Ltd. All rights reserved.

Enabling aqueous processing for crack-free thick electrodes

DOE PAGES

Du, Zhijia; Rollag, K. M.; Li, J.; ...

2017-04-14

Aqueous processing of thick electrodes for Li-ion cells promises to increase energy density due to increased volume fraction of active materials, and to reduce cost due to the elimination of the toxic solvents. Here in this paper this work reports the processing and characterization of aqueous processed electrodes with high areal loading and associated full pouch cell performance. Cracking of the electrode coatings becomes a critical issue for aqueous processing of the positive electrode as areal loading increases above 20–25 mg/cm 2 (~4 mAh/cm 2). Crack initiation and propagation, which was observed during drying via optical microscopy, is related tomore » the build-up of capillary pressure during the drying process. The surface tension of water was reduced by the addition of isopropyl alcohol (IPA), which led to improved wettability and decreased capillary pressure during drying. The critical thickness (areal loading) without cracking increased gradually with increasing IPA content. The electrochemical performance was evaluated in pouch cells. Electrodes processed with water/IPA (80/20 wt%) mixture exhibited good structural integrity with good rate performance and cycling performance.« less

Graphene Oxide Hybridized nHAC/PLGA Scaffolds Facilitate the Proliferation of MC3T3-E1 Cells

NASA Astrophysics Data System (ADS)

Liang, Chunyong; Luo, Yongchao; Yang, Guodong; Xia, Dan; Liu, Lei; Zhang, Xiaomin; Wang, Hongshui

2018-01-01

Biodegradable porous biomaterial scaffolds play a critical role in bone regeneration. In this study, the porous nano-hydroxyapatite/collagen/poly(lactic-co-glycolic acid)/graphene oxide (nHAC/PLGA/GO) composite scaffolds containing different amount of GO were fabricated by freeze-drying method. The results show that the synthesized scaffolds possess a three-dimensional porous structure. GO slightly improves the hydrophilicity of the scaffolds and reinforces their mechanical strength. Young's modulus of the 1.5 wt% GO incorporated scaffold is greatly increased compared to the control sample. The in vitro experiments show that the nHAC/PLGA/GO (1.5 wt%) scaffolds significantly cell adhesion and proliferation of osteoblast cells (MC3T3-E1). This present study indicates that the nHAC/PLGA/GO scaffolds have excellent cytocompatibility and bone regeneration ability, thus it has high potential to be used as scaffolds in the field of bone tissue engineering.

Adsorption of rare earth ions onto the cell walls of wild-type and lipoteichoic acid-defective strains of Bacillus subtilis.

PubMed

Moriwaki, Hiroshi; Koide, Remi; Yoshikawa, Ritsuko; Warabino, Yuya; Yamamoto, Hiroki

2013-04-01

The aim of this study is to investigate the potential of cell walls of wild-type and lipoteichoic acid-defective strains of Bacillus subtilis 168 to adsorb rare earth ions. Freeze-dried cell powders prepared from both strains were used for the evaluation of adsorption ability for the rare earth ions, namely, La(III), Eu(III), and Tm(III). The rare earth ions were efficiently adsorbed onto powders of both wild-type strain (WT powder) and lipoteichoic acid-defective strain (∆LTA powder) at pH 3. The maximum adsorption capacities for Tm(III) by WT and ∆LTA powders were 43 and 37 mg g(-1), respectively. Removal (in percent) of Tm(III), La(III), and Eu(III) from aqueous solution by WT powder was greater than by ∆LTA powder. These results indicate that rare earth ions are adsorbed to functional groups, such as phosphate and carboxyl groups, of lipoteichoic acid. We observed coagulated ∆LTA powder in the removal of rare earth ions (1-20 mg L(-1)) from aqueous solution. In contrast, sedimentation of WT powder did not occur under the same conditions. This unique feature of ∆LTA powder may be caused by the difference of the distribution between lipoteichoic acid and wall teichoic acid. It appears that ∆LTA powder is useful for removal of rare earth ions by adsorption, because aggregation allows for rapid separation of the adsorbent by filtration.

Oxygen requirement for denitrification by the fungus Fusarium oxysporum.

PubMed

Zhou, Z; Takaya, N; Sakairi, M A; Shoun, H

2001-01-01

The effects of dioxygen (O2) on the denitrification activity of the fungus Fusarium oxysporum MT-811 in fed-batch culture in a stirred jar fermentor were examined. The results revealed that fungal denitrifying activity requires a minimal amount of O2 for induction, which is repressed by excess O2. The optimal O2 supply differed between the denitrification substrates : 690 micromol O2 x h(-1) (g dry cell wt.)(-1) for nitrate (NO3-) and about 250 micromol O2 x h(-1) (g dry cell wt.)(-1) for nitrite (NO2-). The reduction of NO3- required more O2 than that of NO2- . With an optimal O2 supply, 80% and 52% of nitrogen atoms in NO3- and NO2-, respectively, were recovered as the denitrification product N2O. These features of F. oxysporum differ from those of bacterial denitrifiers that work exclusively under anoxic conditions. The denitrification activity of F. oxysporum MT-811 mutants with impaired NO3- assimilation was about double that of the wild-type strain, suggesting competition for the substrate between assimilatory and dissimilatory types of NO3- reduction. These results showed that denitrification by F. oxysporum has unique features, namely, a minimal O2 requirement and competition with assimilatory NO3-.

Fabrication of individual alginate-TCP scaffolds for bone tissue engineering by means of powder printing.

PubMed

Castilho, Miguel; Rodrigues, Jorge; Pires, Inês; Gouveia, Barbara; Pereira, Manuel; Moseke, Claus; Groll, Jürgen; Ewald, Andrea; Vorndran, Elke

2015-01-06

The development of polymer-calcium phosphate composite scaffolds with tailored architectures and properties has great potential for bone regeneration. Herein, we aimed to improve the functional performance of brittle ceramic scaffolds by developing a promising biopolymer-ceramic network. For this purpose, two strategies, namely, direct printing of a powder composition consisting of a 60:40 mixture of α/β-tricalcium phosphate (TCP) powder and alginate powder or vacuum infiltration of printed TCP scaffolds with an alginate solution, were tracked. Results of structural characterization revealed that the scaffolds printed with 2.5 wt% alginate-modified TCP powders presented a uniformly distributed and interfusing alginate TCP network. Mechanical results indicated a significant increase in strength, energy to failure and reliability of powder-modified scaffolds with an alginate content in the educts of 2.5 wt% when compared to pure TCP, as well as to TCP scaffolds containing 5 wt% or 7.5 wt% in the educts, in both dry and wet states. Culture of human osteoblast cells on these scaffolds also demonstrated a great improvement of cell proliferation and cell viability. While in the case of powder-mixed alginate TCP scaffolds, isolated alginate gels were formed between the calcium phosphate crystals, the vacuum-infiltration strategy resulted in the covering of the surface and internal pores of the TCP scaffold with a thin alginate film. Furthermore, the prediction of the scaffolds' critical fracture conditions under more complex stress states by the applied Mohr fracture criterion confirmed the potential of the powder-modified scaffolds with 2.5 wt% alginate in the educts as structural biomaterial for bone tissue engineering.

Skeletal phenotype of growing transgenic mice that express a function-perturbing form of beta1 integrin in osteoblasts

NASA Technical Reports Server (NTRS)

Globus, R. K.; Amblard, D.; Nishimura, Y.; Iwaniec, U. T.; Kim, J-B; Almeida, E. A. C.; Damsky, C. D.; Wronski, T. J.; van der Meulen, M. C. H.

2005-01-01

Skeletal modeling entails the deposition of large amounts of extracellular matrix (ECM) to form structures tailored to withstand increasing mechanical loads during rapid growth. Specific ECM molecules bind to integrin receptors on the cell surface, thereby triggering a cascade of signaling events that affect critical cell functions. To evaluate the role of integrins during skeletal growth, transgenic mice were engineered to express a function-perturbing fragment of beta1 integrin consisting of the transmembrane domain and cytoplasmic tail under the control of the osteocalcin promoter (TG mice). Thus, transgene expression was targeted to mature cells of the osteoblast lineage, and herein we show that cultured cells resembling osteocytes from 90-day-old TG mice display impaired adhesion to collagen I, a ligand for beta1 integrin. To determine the influence of beta1 integrin on bones that are responsible for providing structural support during periods of rapid growth, we examined the phenotype of the appendicular skeleton in TG mice compared to wild type (WT) mice. According to radiographs, bones from mice of both genotypes between 14 and 90 days of age appeared similar in gross structure and density, although proximal tibiae from 35-90 days old TG mice were less curved than those of WT mice (72-92% TG/WT). Although there were only mild and transient differences in absolute bone mass and strength, once normalized to body mass, the tibial dry mass (79.1% TG/WT females), ash mass (78.5% TG/WT females), and femoral strength in torsion (71.6% TG/WT females) were reduced in TG mice compared to WT mice at 90 days of age. Similar effects of genotype on bone mass and curvature were observed in 1-year-old retired breeders, indicating that these phenotypic differences between TG and WT mice were stable well into adulthood. Effects of genotype on histomorphometric indices of cancellous bone turnover were minimal and evident only transiently during growth, but when present they demonstrated differences in osteoblast rather than osteoclast parameters. Together, these results suggest that integrin signals generated during growth enhance the acquisition of a skeletal mass, structure, and strength to withstand the mechanical loads generated by weight-bearing.

Spatial distribution of polybrominated diphenyl ethers and polychlorinated dibenzo-p-dioxins and dibenzofurans in soil and combusted residue at Guiyu, an electronic waste recycling site in southeast China.

PubMed

Leung, Anna O W; Luksemburg, William J; Wong, Anthony S; Wong, Ming H

2007-04-15

Surface soils and combusted residue from a village located in southeast China, which has been intensely involved in the dismantling and "recycling" of computer parts (e-waste) for the past decade, were analyzed for polybrominated diphenyl ethers (PBDEs) and polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs). Total PBDE concentrations were highest in combusted residue of plastic chips and cables collected from a residential area (33,000-97,400 ng/g, dry wt), in soils from an acid leaching site (2720-4250 ng/g, dry wt), and a printer roller dump site (593-2890 ng/g, dry wt). BDE-209 was the most dominant congener (35-82%) among the study sites indicating the prevalence of commercial Deca-BDE, however signature congeners from commercial Penta- and Octa-BDE were also found. PCDD/F concentrations were also highest in soil from the acid leaching site (12,500-89,800 pg/g, 203-1100 pg WHO-TEQ/g, dry wt) and in combusted residue (13,500-25,300 pg/g, 84.3-174 pg WHO-TEQ/g, dry wt) and were comparable to PCDD/F levels of some open dumping sites in Asian developing countries. Of the e-waste activities, acid leaching and open burning emitted the highest concentrations of PBDEs and PCDD/Fs. This study is among the very few studies dealing with the important issue of pollution generated from crude e-waste recycling. Our results showthatthe crude processing of e-waste has become one of the major contributors of PBDEs and PCDD/Fs to the terrestrial environment.

Manganese content of soy or rice beverages is high in comparison to infant formulas.

PubMed

Cockell, Kevin A; Bonacci, Giuseppe; Belonje, Bartholomeus

2004-04-01

Well-meaning but inadequately informed parents may perceive plant-based beverages such as soy beverages (SB) or rice beverages (RB) as an alternative to infant formula. Manganese (Mn) is an essential mineral nutrient found at high levels in plants such as soy and rice. Excessive Mn exposure increases the risk of adverse neurological effects. We analysed, by atomic absorption spectrometry, the Mn content of 36 SB, 5 RB, 6 evaporated milks (EM), 14 soy-based infant formulas (SF) and 16 milk-based infant formulas (MF), obtained from commercial outlets in Ottawa, Canada. SB had the highest levels of Mn (16.5 +/- 8.6 micro g/g dry wt, mean +/- s.d.), followed by RB (9.9 +/- 1.7 micro g/g dry wt). Mn levels of individual SB/RB ranged from 2 to 17 times the mean Mn content of SF (2.4 +/- 0.7 micro g/g dry wt) and 7 to 56 times that of MF (0.70 +/- 0.35 micro g/g dry wt). EM contained very little Mn (0.02 +/- 0.03 micro g/g dry wt). Calculated mean Mn intakes from SB/RB by infants up to 6 months of age, assuming complete substitution of these products (0.78 L/day), approached the Tolerable Upper Intake Level (UL) for 1-3 year olds (no UL for Mn is available for infants under 1 year of age). Expressed as micro g Mn/100 kcal, SB/RB exceeded the range derived from ULs and typical energy intakes of 1-3 year olds. SB/RB should not be fed to infants because they are nutritionally inadequate and contain Mn at levels which may present an increased risk of adverse neurological effects if used as a sole source of nutrition.

Mucin deficiency causes functional and structural changes of the ocular surface.

PubMed

Floyd, Anne M; Zhou, Xu; Evans, Christopher; Rompala, Olivia J; Zhu, Lingxiang; Wang, Mingwu; Chen, Yin

2012-01-01

MUC5AC is the most abundant gel-forming mucin in the ocular system. However, the specific function is unknown. In the present study, a Muc5ac knockout (KO) mouse model was subject to various physiological measurements as compared to its wide-type (WT) control. Interestingly, when KO mice were compared to WT mice, the mean tear break up time (TBUT) values were significantly lower and corneal fluorescein staining scores were significantly higher. But the tear volume was not changed. Despite the lack of Muc5ac expression in the conjunctiva of KO mice, Muc5b expression was significantly increased in these mice. Corneal opacification, varying in location and severity, was found in a few KO mice but not in WT mice. The present results suggest a significant difference in the quality, but not the quantity, of tear fluid in the KO mice compared to WT mice. Dry eye disease is multifactorial and therefore further evaluation of the varying components of the tear film, lacrimal unit and corneal structure of these KO mice may help elucidate the role of mucins in dry eye disease. Because Muc5ac knockout mice have clinical features of dry eye, this mouse model will be extremely useful for further studies regarding the pathophysiology of the ocular surface in dry eye in humans.

Nano spray-dried sodium chloride and its effects on the microbiological and sensory characteristics of surface-salted cheese crackers.

PubMed

Moncada, Marvin; Astete, Carlos; Sabliov, Cristina; Olson, Douglas; Boeneke, Charles; Aryana, Kayanush J

2015-09-01

Reducing particle size of salt to approximately 1.5 µm would increase its surface area, leading to increased dissolution rate in saliva and more efficient transfer of ions to taste buds, and hence, perhaps, a saltier perception of foods. This has a potential for reducing the salt level in surface-salted foods. Our objective was to develop a salt using a nano spray-drying method, to use the developed nano spray-dried salt in surface-salted cheese cracker manufacture, and to evaluate the microbiological and sensory characteristics of cheese crackers. Sodium chloride solution (3% wt/wt) was sprayed through a nano spray dryer. Particle sizes were determined by dynamic light scattering, and particle shapes were observed by scanning electron microscopy. Approximately 80% of the salt particles produced by the nano spray dryer, when drying a 3% (wt/wt) salt solution, were between 500 and 1,900 nm. Cheese cracker treatments consisted of 3 different salt sizes: regular salt with an average particle size of 1,500 µm; a commercially available Microsized 95 Extra Fine Salt (Cargill Salt, Minneapolis, MN) with an average particle size of 15 µm; and nano spray-dried salt with an average particle size of 1.5 µm, manufactured in our laboratory and 3 different salt concentrations (1, 1.5, and 2% wt/wt). A balanced incomplete block design was used to conduct consumer analysis of cheese crackers with nano spray-dried salt (1, 1.5, and 2%), Microsized salt (1, 1.5, and 2%) and regular 2% (control, as used by industry) using 476 participants at 1wk and 4mo. At 4mo, nano spray-dried salt treatments (1, 1.5, and 2%) had significantly higher preferred saltiness scores than the control (regular 2%). Also, at 4mo, nano spray-dried salt (1.5 and 2%) had significantly more just-about-right saltiness scores than control (regular 2%). Consumers' purchase intent increased by 25% for the nano spray-dried salt at 1.5% after they were notified about the 25% reduction in sodium content of the cheese cracker. We detected significantly lower yeast counts for nano spray-dried salt treatments (1, 1.5, and 2%) at 4mo compared with control (regular) salt (1, 1.5 and 2%). We detected no mold growth in any of the treatments at any time. At 4mo, we found no significant differences in sensory color, aroma, crunchiness, overall liking, or acceptability scores of cheese crackers using 1.5 and 1% nano spray-dried salt compared with control. Therefore, 25 to 50% less salt would be suitable for cheese crackers if the particle size of regular salt was reduced 3 log to form nano spray-dried salt. A 3-log reduction in sodium chloride particle size from regular salt to nano spray-dried salt increased saltiness, but a 1-log reduction in salt size from Microsized salt to nano spray-dried salt did not increase saltiness of surface-salted cheese crackers. The use of salt with reduced particle size by nano spray drying is recommended for use in surface-salted cheese crackers to reduce sodium intake. Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Fungal melanins and their interactions with metals.

PubMed

Fogarty, R V; Tobin, J M

1996-09-01

Fungal melanins are dark brown or black pigments located in cell walls. They also exist as extracellular polymers. Melanized fungi possess increased virulence and resistance to microbial attack as well as enhanced survival while under environmental stress. Melanins contain various functional groups which provide an array of multiple nonequivalent binding sites for metal ions. Pigmented Cladosporium cladosporoides was shown to biosorb 2.5- to four-fold more Ni, Cu, Zn, Cd, and Pb than albino Penicillium digitatum and at four- to six-fold higher rates. Metal desorption was significantly lower for extracellular melanin than from pigmented or albino biomass which indicated the strength of the melanin-metal bond. At equilibrium, tributyltin chloride (TBTC) concentrations of 2.5 mM, pigmented and albino Aureobasidium pullulans absorbed approximately 0.9 and 0.7 mumol TBTC mg -1 dry wt, respectively, whereas purified extracellular melanin exhibited uptake levels of approximately 22 mumol TBTC mg-1 dry wt at an equilibrium concentration of only 0.4 mM. Addition of melanin to the growth medium reduced the toxic effect of CuSO4 and TBTC due to melanin metal binding and sequestration.

Mass Balance Analysis of Contaminated Heparin Product

PubMed Central

Liu, Zhenling; Xiao, Zhongping; Masuko, Sayaka; Zhao, Wenjing; Sterner, Eric; Bansal, Vinod; Fareed, Jawed; Dordick, Jonathan; Zhang, Fuming; Linhardt, Robert J.

2011-01-01

A quantitative analysis of a recalled contaminated lot of heparin (HP) sodium injection United States Pharmacopeial (USP) was undertaken in response to the controversy regarding the exact nature of the contaminant involved in the HP crisis. A mass balance analysis of the formulated drug product was performed. After freeze-drying, a 1 ml vial for injection afforded 54.8 ± 0.3 mg of dry solids. The excipients, sodium chloride and residual benzyl alcohol, accounted for 11.4 ± 0.5 mg and 0.9 ± 0.5 mg, respectively. Active pharmaceutical ingredient (API) represented 41.5 ± 1.0 mg, corresponding to 75.7 wt% of dry mass. Exhaustive treatment of API with specific enzymes, heparin lyases and/or chondroitin lyases was used to close mass balance. HP represented 30.5 ± 0.5 mg, corresponding to 73.5 wt% of the API. Dermatan sulfate (DS) impurity represented 1.7 ± 0.3 mg, corresponding to 4.1 wt% of the API. Contaminant, 9.3 ± 0.1 mg corresponding to 22.4 wt% of API, was found in the contaminated formulated drug product. The recovery of contaminant was close to quantitative (95.6-100 wt%). A single contaminant was unambiguously identified as oversulfated chondroitin sulfate (OSCS). PMID:20850409

Finger millet (Eleusine coracana) - an economically viable source for antihypercholesterolemic metabolites production by Monascus purpureus.

PubMed

Venkateswaran, V; Vijayalakshmi, G

2010-08-01

Rice, parboiled rice, finger millet, germinated finger millet, broken wheat, njavara (medicinal rice), sorghum and maize were used as substrates for solid state fermentation of Monascus purpureus at 28°C for 7 days using 2% seed medium as inoculum for the production of its metabolites. The fungus exhibited good growth in all the substrates. The fermented substrates were dried at 45°C and analysed for antihypercholesterolemic metabolite statins by standardized HPLC method and dietary sterol contents by spectrophotometric method using reference standards of statin (pravastatin and lovastatin) and cholesterol, respectively. Germinated finger millet yielded higher total statin production of 5.2 g/kg dry wt with pravastatin and lovastatin content of 4.9 and 0.37 g/kg dry wt respectively than other substrates which range from 1.04-4.41 g/kg. In addition to statin, monascus fermented germinated finger millet yielded dietary sterol of 0.053 g/kg dry wt which is 7.6 folds higher than the control. The value addition of finger millet by germination and fermentation with Monascus purpureus provides scope for development of functional food.

Immobilization of Acetobacter aceti on cellulose ion exchangers: adsorption isotherms

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bar, R.; Gainer, J.L.; Kirwan, D.J.

1986-08-01

The adsorptive behavior of cells of Acetobacter aceti, ATCC 23746, on DEAE-, TEAE-, and DEHPAE-cellulose ion exchangers in a modified Hoyer's medium at 30 degrees Centigrade was investigated. The maximum observed adsorption capacities varied from 46 to 64 mg dry wt/g resin. The Langmuir isotherm form was used to fit the data, since the cells formed a monolayer on the resin and exhibited saturation. The equilibrium constant in the Langmuir expression was qualitatively correlated with the surface charge density of the resin. The adsorption was also ''normalized'' by considering the ionic capacities of the resins. The exceptionally high normalized adsorptionmore » capacity of ECTEOLA-cellulose, 261 mg dry/meq, may be explained by an interaction between the cell wall and the polyglyceryl chains of the exchanging groups in addition to the electrostatic effects. The effect of pH on the bacterial adsorption capacity of ECTEOLA-, TEAE-, and phosphate-cellulose resins was studied and the pH of the bacteria was estimated to be 3.0. 17 references.« less

Evaluation of Cation Migration in Lanthanum Strontium Cobalt Ferrite Solid Oxide Fuel Cell Cathodes via In-operando X-ray Diffraction

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hardy, John S.; Coyle, Christopher A.; Bonnett, Jeff F.

2018-01-28

Anode-supported SOFCs with LSCF-6428 cathodes were operated at various temperatures for hundreds of hours in dry or humid (~3% water) cathode air with continuous cathode XRD. Additionally, one cell in dry air was held at OCV and another had 12% CO2 added to the humid air. Long cumulative XRD count times allowed identification of minor phases at <0.1 wt%. In humid air, performance improved during the first couple of hundred hours and Fe-rich Fe,Co spinel XRD peaks gradually shifted to lower angles while nano-nodules formed on LSCF surfaces. With 12% CO2 added, performance degraded after initial activation, unlike without CO2,more » where stability followed activation. In CO2, LSCF XRD peaks shifted indicating gradual decomposition. In dry air, fast initial degradation that decelerated over time occurred at constant current while the cell at OCV was stable. At OCV and 750°C or at constant current and 700°C in dry air, Fe-rich spinel XRD peaks shifted more slowly than in humid air tests; Co-rich Fe,Co spinel peaks shifted to higher angles; and SEM discovered smaller nano-nodules on LSCF than after humid air tests. At constant current at 750°C and 800°C in dry air, no nano-nodules or gradual changes in the XRD patterns were discovered.« less

Nitrate removal performance of Diaphorobacter nitroreducens using biodegradable plastics as the source of reducing power

NASA Astrophysics Data System (ADS)

Khan, S. T.; Nagao, Y.; Hiraishi, A.

2015-02-01

Strain NA10BT and other two strains of the denitrifying betaproteobacterium Diaphorobacter nitroreducens were studied for the performance of solid-phase denitrification (SPD) using poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) and some other biodegradable plastics as the source of reducing power in wastewater treatment. Sequencing-batch SPD reactors with these organisms and PHBV granules or flakes as the substrate exhibited good nitrate removal performance. Vial tests using cultures from these parent reactors showed higher nitrate removal rates with PHBV granules (ca. 20 mg-NO3-- N g-1 [dry wt cells] h-1) than with PHBV pellets and flakes. In continuous-flow SPD reactors using strain NA10BT and PHBV flakes, nitrate was not detected even at a loading rate of 21 mg-NO3-- N L-1 h-1. This corresponded to a nitrate removal rate of 47 mg-NO3-- N g-1 (dry wt cells) h-1. In the continuous-flow reactor, the transcription level of the phaZ gene, coding for PHB depolymerase, decreased with time, while that of the nosZ gene, involved in denitrificaiton, was relatively constant. These results suggest that the bioavailability of soluble metabolites as electron donor and carbon sources increases with time in the continuous-flow SPD process, thereby having much higher nitrate removal rates than the process with fresh PHBV as the substrate.

A survey of antifoulants in sediments from Ports and Marinas along the French Mediterranean coast.

PubMed

Cassi, Roberto; Tolosa, Imma; de Mora, Stephen

2008-11-01

Due to deleterious effects on non-target organisms, the use of organotin compounds on boat hulls of small vessels (<25 m) has been widely prohibited. The International Maritime Organisation (IMO) resolved that the complete prohibition on organotin compounds acting as biocides in antifouling systems should commence in 2008. As a result of restrictions on the use of organotin based paints, other antifouling formulations containing organic biocides have been utilised. This survey was conducted to assess the contamination of replacement biocides in the marine environment following the ban of TBT-based paints. Surface sediments samples were collected in the major ports and marinas along the France Mediterranean coastline (Cote d'Azur) and analysed for organotin compounds, Irgarol 1051, Sea-nine 211, Chlorothalonil, Dichlofluanid and Folpet. Every port and marina exhibited high levels of organotin compounds, with concentrations in sediments ranging from 37 ng Sn g(-1) dry wt in Menton Garavan to over 4000 ng Sn g(-1) dry wt close to the ship chandler within the port of Villefranche-sur-Mer. TBT degradation indexes suggested that fresh inputs are still made. Among the other antifoulants monitored, only Irgarol 1051 exhibited measurable concentrations in almost every port, with concentrations ranging from 40 ng g(-1) dry wt (Cannes) to almost 700 ng g(-1) dry wt (Villefranche-sur-Mer, ship chandler).

Seasonal variations in biomass and species composition of seaweeds along the northern coasts of Persian Gulf (Bushehr Province)

NASA Astrophysics Data System (ADS)

Dadolahi-Sohrab, A.; Garavand-Karimi, M.; Riahi, H.; Pashazanoosi, H.

2012-02-01

This study was carried out to evaluate the seasonal variations of seaweed biomass and species composition at six different sites along the coastal areas in Bushehr Province. Sampling depths varied among sites, from 0.3 to 2.0 m below mean sea level. A total of 37 (i.e., 10 Chlorophyta, 12 Phaeophyta and 15 Rhodophyta) seaweed species were collected. Studies were conducted for quantifying the seaweeds during four seasons from October 2008 until July 2009. During present research, Ulva intestinalis and Cladophora nitellopsis of green, Polycladia myrica, Sirophysalia trinodis and Sargassum angustifolium of brown and Gracilaria canaliculata and Hypnea cervicornis of red seaweeds showed highest biomass in coastal areas of Bushehr Province. The Cheney`s ratio of 2.1 indicated a temperate algal flora to this area. All sites exhibited more than 50% similarity of algal species, indicating a relatively homogenous algal distribution. Total biomass showed the highest value of 3280.7 ± 537.8 g dry wt m - 2 during summer and lowest value of 856.9 ± 92.0 g dry wt m - 2 during winter. During this study, the highest and lowest seaweed biomass were recorded on the site 2 (2473.7 ± 311.0 g dry wt m - 2) and site 5 (856.7 ± 96.8 g dry wt m - 2), respectively.

Mucin Deficiency Causes Functional and Structural Changes of the Ocular Surface

PubMed Central

Evans, Christopher; Rompala, Olivia J.; Zhu, Lingxiang; Wang, Mingwu; Chen, Yin

2012-01-01

MUC5AC is the most abundant gel-forming mucin in the ocular system. However, the specific function is unknown. In the present study, a Muc5ac knockout (KO) mouse model was subject to various physiological measurements as compared to its wide-type (WT) control. Interestingly, when KO mice were compared to WT mice, the mean tear break up time (TBUT) values were significantly lower and corneal fluorescein staining scores were significantly higher. But the tear volume was not changed. Despite the lack of Muc5ac expression in the conjunctiva of KO mice, Muc5b expression was significantly increased in these mice. Corneal opacification, varying in location and severity, was found in a few KO mice but not in WT mice. The present results suggest a significant difference in the quality, but not the quantity, of tear fluid in the KO mice compared to WT mice. Dry eye disease is multifactorial and therefore further evaluation of the varying components of the tear film, lacrimal unit and corneal structure of these KO mice may help elucidate the role of mucins in dry eye disease. Because Muc5ac knockout mice have clinical features of dry eye, this mouse model will be extremely useful for further studies regarding the pathophysiology of the ocular surface in dry eye in humans. PMID:23272068

Temporal and spatial distribution of phytoplankton functional groups and role of environment factors in a deep subtropical reservoir

NASA Astrophysics Data System (ADS)

Li, Lei; Li, Qiuhua; Chen, Jing'an; Wang, Jingfu; Jiao, Shulin; Chen, Fengfeng

2017-09-01

Phytoplankton and environment factors were investigated in 2015 and phytoplankton functional groups were used to understand their temporal and spatial distribution and their driving factors in Wanfeng Reservoir. Seventeen functional groups (B, D, E, F, G, J, Lo, MP, P, S1, T, W1, W2, X1, X2, Xph, Y) were identified based on 34 species. The dominant groups were: J/B/P/D in dry season, X1/J/Xph/G/T in normal season and J in flood season. Phytoplankton abundance ranged from 5.33×104 cells/L to 3.65×107 cells/L, with the highest value occurring in flood season and lowest in dry season. The vertical profile of dominant groups showed little differentiation except for P, which dominated surface layers over 20 m as a result of mixing water masses and higher transparency during dry season. However, the surface waters presented higher values of phytoplankton abundance than other layers, possibly because of greater irradiance. The significant explaining variables and their ability to describe the spatial distribution of the phytoplankton community in RDA differed seasonally as follows: dry season, NH4-N, NO3-N, NO2-N, TN:TP ratio and transparency (SD); normal season, temperature (WT), water depth, TN, NH4-N and NO3-N; flood season, WT, water depth, NO3-N and NO2-N. Furthermore, nitrogen, water temperature, SD and water depth were significant variables explaining the variance of phytoplankton communities when datasets included all samples. The results indicated that water physical conditions and hydrology were important in phytoplankton community dynamics, and nitrogen was more important than phosphorus in modifying phytoplankton communities. Seasonal differences in the relationship between the environment and phytoplankton community should be considered in water quality management.

The effects of composition and sintering temperature on the silica foam fabricated by slurry method

NASA Astrophysics Data System (ADS)

Baharom, Syazwani; Ahmad, Sufizar; Taib, Hariati; Muda, Rizamarhaiza

2016-07-01

Reticulated ceramic or open pore ceramic foam is a well-known material which exhibits extremely high porosities, with a significant degree of interconnectivity that makes them desirable in a wide range of applications. There were broad types of ceramic foam fabrication method such as polymeric sponge method, direct foaming, and starch consolidation. In this study, the slurry method has been chosen to fabricate Silica (SiO2) foam. In this process, Polyurethane (PU) foam template was dipped into ceramic slurry and followed by drying and sintering to obtain foam which contains porosity in the range of 50% to 70%. The compositions of SiO2 were varied starting from 55 wt.%, 60 wt.%, 65 wt.% and 70 wt.%. The samples of SiO2 that have been dipped and dried were sintered at 900°C, 1000°C, 1100°C, and 1250°C. The sintered SiO2 ceramic foam samples were characterized to observe their morphology, and physical properties. Thus, the microstructure of the SiO2 ceramic foams samples was examined by Scanning Electron Microscopy (SEM), and Electron Dispersive Spectroscopy (EDS). Meanwhile, the physical properties of the SiO2 ceramic foam samples such as the total porosity (%) and bulk density were determined using Archimedes method. It was found that the density of ceramic foam produced was in the range of 0.25 g/cm3 up to 0.75 g/cm3, whereas the level of porosity percentage was in the range of 61.81% to 82.18% with the size of open pore or window cells were in between 141 µm up to 626 µm.

The effects of composition and sintering temperature on the silica foam fabricated by slurry method

DOE Office of Scientific and Technical Information (OSTI.GOV)

Baharom, Syazwani, E-mail: hd140001@siswa.uthm.edu.my; Ahmad, Sufizar, E-mail: sufizar@uthm.edu.my; Taib, Hariati, E-mail: hariati@uthm.edu.my

Reticulated ceramic or open pore ceramic foam is a well-known material which exhibits extremely high porosities, with a significant degree of interconnectivity that makes them desirable in a wide range of applications. There were broad types of ceramic foam fabrication method such as polymeric sponge method, direct foaming, and starch consolidation. In this study, the slurry method has been chosen to fabricate Silica (SiO{sub 2}) foam. In this process, Polyurethane (PU) foam template was dipped into ceramic slurry and followed by drying and sintering to obtain foam which contains porosity in the range of 50% to 70%. The compositions ofmore » SiO{sub 2} were varied starting from 55 wt.%, 60 wt.%, 65 wt.% and 70 wt.%. The samples of SiO{sub 2} that have been dipped and dried were sintered at 900°C, 1000°C, 1100°C, and 1250°C. The sintered SiO{sub 2} ceramic foam samples were characterized to observe their morphology, and physical properties. Thus, the microstructure of the SiO{sub 2} ceramic foams samples was examined by Scanning Electron Microscopy (SEM), and Electron Dispersive Spectroscopy (EDS). Meanwhile, the physical properties of the SiO{sub 2} ceramic foam samples such as the total porosity (%) and bulk density were determined using Archimedes method. It was found that the density of ceramic foam produced was in the range of 0.25 g/cm{sup 3} up to 0.75 g/cm{sup 3}, whereas the level of porosity percentage was in the range of 61.81% to 82.18% with the size of open pore or window cells were in between 141 µm up to 626 µm.« less

Evaluation of different solvent mixtures in esterifiable lipids extraction from microalgae Botryococcus braunii for biodiesel production.

PubMed

Hidalgo, Pamela; Ciudad, Gustavo; Navia, Rodrigo

2016-02-01

Non-polar and polar solvents as well as their mixtures were tested for the extraction of microalgae lipids and thus, to evaluate their effect on total and esterifiable lipids extraction yields with potential to be converted to biodiesel. The obtained results show an increase in lipids and esterifiable lipids extraction yields when non-polar and polar solvent mixtures were used. The higher esterifiable lipids extraction yield was 19.2%wt (based on dry biomass) using a chloroform-methanol mixture (75%v/v of methanol), corresponding to a 98.9%wt esterifiable lipids extraction. In addition, esterifiable lipids extraction yield of 18.9%wt (based on dry biomass) was obtained when a petroleum ether-methanol mixture (75%v/v of methanol) was used, corresponding to a 96.9%wt esterifiable lipids extraction. Copyright © 2015 Elsevier Ltd. All rights reserved.

Use of electron microprobe x-ray analysis for determination of low calcium concentrations across leaves deficient in calcium

NASA Technical Reports Server (NTRS)

Barta, D. J.; Tibbitts, T. W.

1991-01-01

An electron microprobe with wavelength-dispersive x-ray spectrometry (WDS) was found to be useful for the determination of Ca concentrations in leaf tissue deficient in Ca. WDS effectively detected Ca concentrations as low as 0.2 mg/g dry wt in the presence of high levels of K and Mg (120 and 50 mg/g dry wt, respectively). Leaf specimens were prepared for analysis by quick-freezing in liquid nitrogen and freeze-drying at -20 degrees C to maintain elemental integrity within the tissue. Because dry material was analyzed, sample preparation was simple and samples could be stored for long periods before analysis. A large beam diameter of 50 gm was used to minimize tissue damage under the beam and analyze mineral concentrations within several cells at one time. Beam penetration was between 50 and 55 microns, approximately one-third of the thickness of the leaf. For analysis of concentrations in interveinal areas, analyses directed into the abaxial epidermis were found most useful. However, because of limited beam penetration, analyses of veinal areas would require use of cross sections [correction of crosssections]. Solid mineral standards were used for instrument standardization. To prevent measurement errors resulting from differences between the matrix of the mineral standards and the analyzed tissue, concentrations in leaves were corrected using gelatin standards prepared and analyzed under the same conditions. WDS was found to be useful for documenting that very low Ca levels occur in specific areas of lettuce leaves exhibiting the Ca deficiency injury termed tipburn.

Properties of flat-pressed wood plastic composites containing fire retardants

Treesearch

Nadir Ayrilmis; Jan. T. Benthien; Heiko Thoemen; Robert H. White

2011-01-01

This study investigated physical, mechanical, and fire properties of the flat-pressed wood plastic composites (WPCs) incorporated with various fire retardants (FRs) [5 or 15% by weight (wt)] at 50 wt % of the wood flour (WF). The WPC panels were made from dry-blended WF, polypropylene (PP) with maleic anhydride grafted PP (2 wt %), and FR powder formulations using a...

Soil contamination by brominated flame retardants in open waste dumping sites in Asian developing countries.

PubMed

Eguchi, Akifimi; Isobe, Tomohiko; Ramu, Karri; Tue, Nguyen Minh; Sudaryanto, Agus; Devanathan, Gnanasekaran; Viet, Pham Hung; Tana, Rouch Seang; Takahashi, Shin; Subramanian, Annamalai; Tanabe, Shinsuke

2013-03-01

In Asian developing countries, large amounts of municipal wastes are dumped into open dumping sites each day without adequate management. This practice may cause several adverse environmental consequences and increase health risks to local communities. These dumping sites are contaminated with many chemicals including brominated flame retardants (BFRs) such as polybrominated diphenyl ethers (PBDEs) and hexabromocyclododecanes (HBCDs). BFRs may be released into the environment through production processes and through the disposal of plastics and electronic wastes that contain them. The purpose of this study was to elucidate the status of BFR pollution in municipal waste dumping sites in Asian developing countries. Soil samples were collected from six open waste dumping sites and five reference sites in Cambodia, India, Indonesia, Malaysia, and Vietnam from 1999 to 2007. The results suggest that PBDEs are the dominant contaminants in the dumping sites in Asian developing countries, whereas HBCD contamination remains low. Concentrations of PBDEs and HBCDs ranged from ND to 180 μg/kg dry wt and ND to 1.4 μg/kg dry wt, respectively, in the reference sites and from 0.20 to 430 μg/kg dry wt and ND to 2.5 μg/kg dry wt, respectively, in the dumping sites. Contamination levels of PBDEs in Asian municipal dumping sites were comparable with those reported from electronic waste dismantling areas in Pearl River delta, China. Copyright © 2012 Elsevier Ltd. All rights reserved.

Ultrafine cementitious grout

DOEpatents

Ahrens, Ernst H.

1999-01-01

An ultrafine cementitious grout in three particle grades containing Portland cement, pumice as a pozzolanic material and superplasticizer in the amounts of about 30 wt. % to about 70 wt. % Portland cement; from about 30 wt. % to about 70 wt. % pumice containing at least 70% amorphous silicon dioxide; and from 1.2 wt. % to about 5.0 wt. % superplasticizer. The superplasticizer is dispersed in the mixing water prior to the addition of dry grout and the W/CM ratio is about 0.4 to 1/1. The grout has very high strength and very low permeability with good workability. The ultrafine particle sizes allow for sealing of microfractures below 10 .mu.m in width.

Recovery of slaughterhouse Animal Fatty Wastewater Sludge by conversion into Fatty Acid Butyl Esters by acid-catalyzed esterification.

PubMed

Wallis, Christopher; Cerny, Muriel; Lacroux, Eric; Mouloungui, Zéphirin

2017-02-01

Two types of Animal Fatty Wastewater Sludges (AFWS 1 and 2) were analyzed and fully characterized to determine their suitability for conversion into biofuel. AFWS 1 was determined to be unsuitable as it contains 68.8wt.% water and only 32.3wt.% dry material, of which only around 80% is lipids to be converted. AFWS 2 has only 15.7wt.% water and 84.3wt.% dry material of which is assumed to 100% lipids as the protein and ash contents were determined to be negligible. The 4-dodecylbenzenesulfonic acid (DBSA) catalyzed esterification of AFWS with 1-butanol was performed in a novel batch reactor fitted with a drying chimney for the "in situ" removal of water and optimized using a non-conventional Doehlert surface response methodology. The optimized condition was found to be 1.66mol equivalent of 1-butanol (with respect to total fatty acid chains), 10wt.% of DBSA catalyst (with respect to AFWS) at 105°C for 3h. Fatty Acid Butyl Esters (FABEs) were isolated in good yields (95%+) as well as a blend of FABEs with 1-butanol (16%). The two potential biofuels were analyzed in comparison with current and analogous biofuels (FAME based biodiesel, and FABE products made from vegetable oils) and were found to exhibit high cetane numbers and flash point values. Copyright © 2016 Elsevier Ltd. All rights reserved.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Khan, S. T.; Nagao, Y.; Hiraishi, A., E-mail: hiraishi@ens.tut.ac.jp

Strain NA10B{sup T} and other two strains of the denitrifying betaproteobacterium Diaphorobacter nitroreducens were studied for the performance of solid-phase denitrification (SPD) using poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) and some other biodegradable plastics as the source of reducing power in wastewater treatment. Sequencing-batch SPD reactors with these organisms and PHBV granules or flakes as the substrate exhibited good nitrate removal performance. Vial tests using cultures from these parent reactors showed higher nitrate removal rates with PHBV granules (ca. 20 mg-NO{sub 3}{sup −}‐N g{sup −1} [dry wt cells] h{sup −1}) than with PHBV pellets and flakes. In continuous-flow SPD reactors using strain NA10B{sup T}more » and PHBV flakes, nitrate was not detected even at a loading rate of 21 mg-NO{sub 3}{sup −}‐N L{sup −1} h{sup −1}. This corresponded to a nitrate removal rate of 47 mg-NO{sub 3}{sup −}‐N g{sup −1} (dry wt cells) h{sup −1}. In the continuous-flow reactor, the transcription level of the phaZ gene, coding for PHB depolymerase, decreased with time, while that of the nosZ gene, involved in denitrificaiton, was relatively constant. These results suggest that the bioavailability of soluble metabolites as electron donor and carbon sources increases with time in the continuous-flow SPD process, thereby having much higher nitrate removal rates than the process with fresh PHBV as the substrate.« less

Changes in Cell Wall Polysaccharides Associated With Growth 1

PubMed Central

Nevins, Donald J.; English, Patricia D.; Albersheim, Peter

1968-01-01

Changes in the polysaccharide composition of Phaseolus vulgaris, P. aureus, and Zea mays cell walls were studied during the first 28 days of seedling development using a gas chromatographic method for the analysis of neutral sugars. Acid hydrolysis of cell wall material from young tissues liberates rhamnose, fucose, arabinose, xylose, mannose, galactose, and glucose which collectively can account for as much as 70% of the dry weight of the wall. Mature walls in fully expanded tissues of these same plants contain less of these constituents (10%-20% of dry wt). Gross differences are observed between developmental patterns of the cell wall in the various parts of a seedling, such as root, stem, and leaf. The general patterns of wall polysaccharide composition change, however, are similar for analogous organs among the varieties of a species. Small but significant differences in the rates of change in sugar composition were detected between varieties of the same species which exhibited different growth patterns. The cell walls of species which are further removed phylogenetically exhibit even more dissimilar developmental patterns. The results demonstrate the dynamic nature of the cell wall during growth as well as the quantitative and qualitative exactness with which the biosynthesis of plant cell walls is regulated. PMID:16656862

Concurrent extraction and reaction for the production of biodiesel from wet microalgae.

PubMed

Im, Hanjin; Lee, HanSol; Park, Min S; Yang, Ji-Won; Lee, Jae W

2014-01-01

This work addresses a reliable in situ transesterification process which integrates lipid extraction from wet microalgae, and its conversion to biodiesel, with a yield higher than 90 wt.%. This process enables single-step production of biodiesel from microalgae by mixing wet microalgal cells with solvent, methanol, and acid catalyst; and then heating them in one pot. The effects of reaction parameters such as reaction temperature, wet cell weight, reaction time, and catalyst volume on the conversion yield are investigated. This simultaneous extraction and transesterification of wet microalgae may enable a significant reduction in energy consumption by eliminating the drying process of algal cells and realize the economic production of biodiesel using wet microalgae. Copyright © 2013 Elsevier Ltd. All rights reserved.

Nanostructured thermoplastic polyimide films

DOEpatents

Aglan, Heshmat

2015-05-19

Structured films containing multi-walled carbon nanotubes ("MWCNTs") have enhanced mechanical performance in terms of strength, fracture resistance, and creep recovery of polyimide ("PI") films. Preferably, the loadings of MWCNTs can be in the range of 0.1 wt % to 0.5 wt %. The strength of the new PI films dried at 60.degree. C. increased by 55% and 72% for 0.1 wt % MWCNT and 0.5 wt % MWCNT loadings, respectively, while the fracture resistance increased by 23% for the 0.1 wt % MWCNTs and then decreases at a loading of 0.5 wt % MWCNTs. The films can be advantageously be created by managing a corresponding shift in the annealing temperature at which the maximum strength occurs as the MWCNT loadings increase.

A comparison of bacterial and fungal biomass in several cultivated soils.

PubMed

Kaczmarek, W

1984-01-01

Bacterial and fungal biomass was estimated in incubated samples of three cultivated soils, the influence of glucose, ammonium nitrate and cattle slurry on its formation being studied. The microbial biomass was determined in stained microscopic preparations of soil suspension. Bacterial biomass in the control samples was from 0.17 to 0.66 mg dry wt per 1 g dry soil and independently of the applied supplements was on the average two times larger in muck soils than in sand. Fungal biomass in the control soils ranged from 0.013 to 0.161 mg dry wt per 1 g dry soil, no relationship being found between its size and the soil type. As a result, the ratio of the size of fungal to bacterial biomass was dependent on the soil type; in sand the fungal biomass corresponded to 1/3 of the bacterial biomass, and in muck soils--only to 1/7.

Effects and risk assessment of linear alkylbenzene sulfonates in agricultural soil. 1. Short-term effects on soil microbiology.

PubMed

Elsgaard, L; Petersen, S O; Debosz, K

2001-08-01

Linear alkylbenzene sulfonates (LAS) may occur in sewage sludge that is applied to agricultural soil, in which LAS can be inhibitory to biological activity. As a part of a broader risk assessment of LAS in the terrestrial environment, we tested the short-term effects of aqueous LAS on microbial parameters in a sandy agricultural soil that was incubated for up to 11 d. The assays included 10 microbial soil parameters; ethylene degradation; potential ammonium oxidation; potential dehydrogenase activity; beta-glucosidase activity; iron reduction; the populations of cellulolytic bacteria, fungi and actinomycetes; the basal soil respiration; and the phospholipid fatty acid (PLFA) content. Except for beta-glucosidase activity, basal respiration, and total PLFA content, all soil parameters were sensitive to LAS, with EC10 values in the range of less than 8 to 22 mg/kg dry weight. This probably reflected a similar mode of LAS toxicity, ascribed to cell membrane interactions, and showed that sensitivity to LAS was common for various soil microorganisms. The extracellular beta-glucosidase activity was rather insensitive to LAS (ECI10, 47 mg/kg dry wt), whereas the basal soil respiration was not inhibited even at 793 mg/kg dry weight. This was interpreted as a combined response of inhibited and stimulated compartments of the microbial community. The PLFA content, surprisingly, showed no decrease even at 488 mg/kg. In conclusion, LAS inhibited specific microbial activities, although this could not be deduced from the basal respiration or the total PLFA content. The lowest EC10 values for microbial soil parameters were slightly higher than the predicted no-effect concentrations recently derived for plants and soil fauna (approximately 5 mg/kg dry wt).

Switchable hydrophilicity solvents for lipid extraction from microalgae for biofuel production.

PubMed

Boyd, Alaina R; Champagne, Pascale; McGinn, Patrick J; MacDougall, Karen M; Melanson, Jeremy E; Jessop, Philip G

2012-08-01

A switchable hydrophilicity solvent (SHS) was studied for its effectiveness at extracting lipids from freeze-dried samples of Botryococcus braunii microalgae. The SHS N,N-dimethylcyclohexylamine extracted up to 22 wt.% crude lipid relative to the freeze-dried cell weight. The solvent was removed from the extract with water saturated with carbon dioxide at atmospheric pressure and recovered from the water upon de-carbonation of the mixture. Liquid chromatography-mass spectrometry (LC-MS) showed that the extracted lipids contained high concentrations of long chain tri-, di- and mono-acylglycerols, no phospholipids, and only 4-8% of residual solvent. Unlike extractions with conventional organic solvents, this new method requires neither distillation nor the use of volatile, flammable or chlorinated organic solvents. Copyright © 2012 Elsevier Ltd. All rights reserved.

Sources and seasonal variation of PAHs in the sediments of drinking water reservoirs in Hong Kong and the Dongjiang River (China).

PubMed

Liang, Yan; Fung, Pui Ka; Tse, Man Fung; Hong, Hua Chang; Wong, Ming Hung

2008-11-01

The main objective of this study was to investigate occurrence of polycyclic aromatic hydrocarbons (PAHs) in the sources of the drinking water supply of Hong Kong. The main emphasis was on the Dongjiang River in mainland China which is the major source, supplying 80% of the total consumption in Hong Kong (the remaining 20% is obtained from rain water). Sediments were collected from four sites along the Dongjiang River and four reservoirs in Hong Kong during both the dry and wet weather seasons. The concentrations of total PAHs in the sediments ranged between 36 and 539 microg/kg dry wt. The lower levels were detected at the upstream site on the Dongjiang River and at the reservoirs in Hong Kong (44-85 microg/kg dry wt), while the mid- and downstream sites on the Dongjiang River were more polluted (588-658 microg/kg dry wt). Examination of the PAH profiles revealed that the mid- and downstream sections of the Dongjiang River contained high percentages of 4,5,6-ring PAHs, similar to the amounts of atmospheric particulate matter and road dust collected during the dry weather season from the Pearl River Delta region as reported in the literature. Seasonal changes were revealed in the reservoirs of Hong Kong, with higher PAH levels in the wet weather season than in the dry weather season. For those reservoirs in Hong Kong that store water from the Dongjiang River, a distinct seasonal pattern was also observed, namely, that under dry weather season conditions the PAHs found in the sediments were primarily from petrogenic source, while under wet weather season conditions they were from pyrolytic sources. No such pattern was detected in the reservoirs which stored only rain water.

Growth characteristics of aquatic macrophytes cultured in nutrient-enriched water. I. Water hyacinth, water lettuce, and pennywort

DOE Office of Scientific and Technical Information (OSTI.GOV)

Reddy, K.R.; DeBusk, W.F.

Seasonal growth characteristics and biomass yield potential of 3 floating aquatic macrophytes cultured in nutrient nonlimiting conditions were evaluated in central Florida's climatic conditions. Growth cycle (growth curve) of the plants was found to be complete when maximum plant density was reached and no additional increase in growth was recorded. Biomass yield per unit area and time was found to be maximum in the linear phase of the growth curve; plant density in this phase was defined as ''operational plant density,'' a density range in which a biomass production system is operated to obtain the highest possible yields. Biomass yieldsmore » were found to be 106, 72, and 41 t(dry wt) ha/sup -1/yr/sup -1/, respectively, for water hyacinth (Eichhornia crassipes), water lettuce (Pistia stratiotes), and pennywort (Hydrocotyle umbellata). Operational plant density was found to be in the range of 500-2000 g dry wt m/sup -2/ for water hyacinth, 200-700 g dry wt m/sup -2/ for water lettuce, and 250-650 g dry wt/sup -2/ for pennywort. Seasonality was observed in growth rates but not in operational plant density. Specific growth rate (% increase per day) was found to maximum at low plant densities and decreased as the plant density increased. Results show that water hyacinth and water lettuce can be successfully grown for a period of about 10 mo, while pennywort, a cool season plant, can be integrated into water hyacinth/water lettuce biomass production system to obtain high yields in the winter.« less

Growth characteristics of aquatic macrophytes cultured in nutrient-enriched water. I. Water hyacinth, water lettuce, and pennywort

DOE Office of Scientific and Technical Information (OSTI.GOV)

Reddy, K.R.; DeBusk, W.F.

Seasonal growth characteristics and biomass yield potential of 3 floating aquatic macrophytes cultured in nutrient nonlimiting conditions were evaluated in central Florida's climatic conditions. Growth cycle (growth curve) of the plants was found to be complete when maximum plant density was reached and no additional increase in growth was recorded. Biomass yield per unit area and time was found to be maximum in the linear phase of the growth curve; plant density in this phase was defined as operational plant density, a density range in which a biomass production system is operated to obtain the highest possible yields. Biomass yieldsmore » were found to be 106, 72, and 41 t (dry wt) ha/sup -1/ yr/sup -1/, respectively, for water hyacinth (Eichhornia crassipes), water lettuce (Pistia stratiotes), and pennywort (Hydrocotyle umbellata). Operational plant density was found to be in the range of 500-2,000 g dry wt m/sup -2/ for water hyacinth, 200-700 g dry wt m/sup -2/ for water lettuce, and 250-650 g dry wt m/sup -2/ for pennywort. Seasonality was observed in growth rates but not in operational plant density. Specific growth rate (% increase per day) was found to maximum at low plant densities and decreased as the plant density increased. Results show that water hyacinth and water lettuce can be successfully grown for a period of about 10 mo, while pennywort, a cool season plant, can be integrated into water hyacinth/water lettuce biomass production system to obtain high yields in the winter.« less

Carbon dioxide exchange in compact and semi-open sorghum inflorescences

DOE Office of Scientific and Technical Information (OSTI.GOV)

Eastin, J.D.; Sullivan, C.Y.

Carbon dioxide exchange rates were monitored in light and dark in compact and semi-open heads of sorghum (Sorghum bicolor (L.) Moench). Developmental stages ranged from bottom to hard dough in the grain. Highest CO/sub 2/ uptake in both head types occurred at the bloom stage when net uptake rates for semi-open and compact type heads were 3.9 and 1.2 mg CO/sub 2/ g dry wt/sup -1/ hr/sup -1/, respectively. Beginning at the milk stage, a net CO/sub 2/ evolution on the order of 1 to 1.4 mg g dry wt/sup -1/ hr/sup -1/ occurred in compact heads in the light.more » The semi-open head type continued a small net CO/sub 2/ uptake in the light through the milk and soft dough stages. Both head types evolved CO/sub 2/ at hard dough stage. Dark respiration was similar in both head types and decreased from about 4 to 1 mg CO/sub 2/ g dry wt/sup -1/ hr/sup -1/ from bloom to hard dough. 16 references, 1 figure.« less

A new high temperature reactor for operando XAS: Application for the dry reforming of methane over Ni/ZrO2 catalyst.

PubMed

Aguilar-Tapia, Antonio; Ould-Chikh, Samy; Lahera, Eric; Prat, Alain; Delnet, William; Proux, Olivier; Kieffer, Isabelle; Basset, Jean-Marie; Takanabe, Kazuhiro; Hazemann, Jean-Louis

2018-03-01

The construction of a high-temperature reaction cell for operando X-ray absorption spectroscopy characterization is reported. A dedicated cell was designed to operate as a plug-flow reactor using powder samples requiring gas flow and thermal treatment at high temperatures. The cell was successfully used in the reaction of dry reforming of methane (DRM). We present X-ray absorption results in the fluorescence detection mode on a 0.4 wt. % Ni/ZrO 2 catalyst under realistic conditions at 750 °C, reproducing the conditions used for a conventional dynamic microreactor for the DRM reaction. The setup includes a gas distribution system that can be fully remotely operated. The reaction cell offers the possibility of transmission and fluorescence detection modes. The complete setup dedicated to the study of catalysts is permanently installed on the Collaborating Research Groups French Absorption spectroscopy beamline in Material and Environmental sciences (CRG-FAME) and French Absorption spectroscopy beamline in Material and Environmental sciences at Ultra-High Dilution (FAME-UHD) beamlines (BM30B and BM16) at the European Synchrotron Radiation Facility in Grenoble, France.

A new high temperature reactor for operando XAS: Application for the dry reforming of methane over Ni/ZrO2 catalyst

NASA Astrophysics Data System (ADS)

Aguilar-Tapia, Antonio; Ould-Chikh, Samy; Lahera, Eric; Prat, Alain; Delnet, William; Proux, Olivier; Kieffer, Isabelle; Basset, Jean-Marie; Takanabe, Kazuhiro; Hazemann, Jean-Louis

2018-03-01

The construction of a high-temperature reaction cell for operando X-ray absorption spectroscopy characterization is reported. A dedicated cell was designed to operate as a plug-flow reactor using powder samples requiring gas flow and thermal treatment at high temperatures. The cell was successfully used in the reaction of dry reforming of methane (DRM). We present X-ray absorption results in the fluorescence detection mode on a 0.4 wt. % Ni/ZrO2 catalyst under realistic conditions at 750 °C, reproducing the conditions used for a conventional dynamic microreactor for the DRM reaction. The setup includes a gas distribution system that can be fully remotely operated. The reaction cell offers the possibility of transmission and fluorescence detection modes. The complete setup dedicated to the study of catalysts is permanently installed on the Collaborating Research Groups French Absorption spectroscopy beamline in Material and Environmental sciences (CRG-FAME) and French Absorption spectroscopy beamline in Material and Environmental sciences at Ultra-High Dilution (FAME-UHD) beamlines (BM30B and BM16) at the European Synchrotron Radiation Facility in Grenoble, France.

Sertoli Cell Wt1 Regulates Peritubular Myoid Cell and Fetal Leydig Cell Differentiation during Fetal Testis Development.

PubMed

Wen, Qing; Wang, Yuqian; Tang, Jixin; Cheng, C Yan; Liu, Yi-Xun

2016-01-01

Sertoli cells play a significant role in regulating fetal testis compartmentalization to generate testis cords and interstitium during development. The Sertoli cell Wilms' tumor 1 (Wt1) gene, which encodes ~24 zinc finger-containing transcription factors, is known to play a crucial role in fetal testis cord assembly and maintenance. However, whether Wt1 regulates fetal testis compartmentalization by modulating the development of peritubular myoid cells (PMCs) and/or fetal Leydig cells (FLCs) remains unknown. Using a Wt1-/flox; Amh-Cre mouse model by deleting Wt1 in Sertoli cells (Wt1SC-cKO) at embryonic day 14.5 (E14.5), Wt1 was found to regulate PMC and FLC development. Wt1 deletion in fetal testis Sertoli cells caused aberrant differentiation and proliferation of PMCs, FLCs and interstitial progenitor cells from embryo to newborn, leading to abnormal fetal testis interstitial development. Specifically, the expression of PMC marker genes α-Sma, Myh11 and Des, and interstitial progenitor cell marker gene Vcam1 were down-regulated, whereas FLC marker genes StAR, Cyp11a1, Cyp17a1 and Hsd3b1 were up-regulated, in neonatal Wt1SC-cKO testes. The ratio of PMC:FLC were also reduced in Wt1SC-cKO testes, concomitant with a down-regulation of Notch signaling molecules Jag 1, Notch 2, Notch 3, and Hes1 in neonatal Wt1SC-cKO testes, illustrating changes in the differentiation status of FLC from their interstitial progenitor cells during fetal testis development. In summary, Wt1 regulates the development of FLC and interstitial progenitor cell lineages through Notch signaling, and it also plays a role in PMC development. Collectively, these effects confer fetal testis compartmentalization.

High-level expression of a synthetic gene encoding a sweet protein, monellin, in Escherichia coli.

PubMed

Chen, Zhongjun; Cai, Heng; Lu, Fuping; Du, Lianxiang

2005-11-01

The expression of a synthetic gene encoding monellin, a sweet protein, in E. coli under the control of T7 promoter from phage is described. The single-chain monellin gene was designed based on the biased codons of E. coli so as to optimize its expression. Monellin was produced and accounted for 45% of total soluble proteins. It was purified to yield 43 mg protein per g dry cell wt. The purity of the recombinant protein was confirmed by SDS-PAGE.

Preparation and Evaluation of Gelatin-Chitosan-Nanobioglass 3D Porous Scaffold for Bone Tissue Engineering

PubMed Central

Maji, Kanchan; Dasgupta, Sudip; Pramanik, Krishna; Bissoyi, Akalabya

2016-01-01

The aim of the present study was to prepare and characterize bioglass-natural biopolymer based composite scaffold and evaluate its bone regeneration ability. Bioactive glass nanoparticles (58S) in the size range of 20–30 nm were synthesized using sol-gel method. Porous scaffolds with varying bioglass composition from 10 to 30 wt% in chitosan, gelatin matrix were fabricated using the method of freeze drying of its slurry at 40 wt% solids loading. Samples were cross-linked with glutaraldehyde to obtain interconnected porous 3D microstructure with improved mechanical strength. The prepared scaffolds exhibited >80% porosity with a mean pore size range between 100 and 300 microns. Scaffold containing 30 wt% bioglass (GCB 30) showed a maximum compressive strength of 2.2 ± 0.1 MPa. Swelling and degradation studies showed that the scaffold had excellent properties of hydrophilicity and biodegradability. GCB 30 scaffold was shown to be noncytotoxic and supported mesenchymal stem cell attachment, proliferation, and differentiation as indicated by MTT assay and RUNX-2 expression. Higher cellular activity was observed in GCB 30 scaffold as compared to GCB 0 scaffold suggesting the fact that 58S bioglass nanoparticles addition into the scaffold promoted better cell adhesion, proliferation, and differentiation. Thus, the study showed that the developed composite scaffolds are potential candidates for regenerating damaged bone tissue. PMID:26884764

Ethylene signaling triggered by low concentrations of ascorbic acid regulates biomass accumulation in Arabidopsis thaliana.

PubMed

Caviglia, M; Mazorra Morales, L M; Concellón, A; Gergoff Grozeff, G E; Wilson, M; Foyer, C H; Bartoli, C G

2018-02-02

Ascorbic acid (AA) is a major redox buffer in plant cells. The role of ethylene in the redox signaling pathways that influence photosynthesis and growth was explored in two independent AA deficient Arabidopsis thaliana mutants (vtc2-1 and vtc2-4). Both mutants, which are defective in the AA biosynthesis gene GDP-L-galactose phosphorylase, produce higher amounts of ethylene than wt plants. In contrast to the wt, the inhibition of ethylene signaling increased leaf conductance, photosynthesis and dry weight in both vtc2 mutant lines. The AA-deficient mutants showed altered expression of genes encoding proteins involved in the synthesis/responses to phytohormones that control growth, particularly auxin, cytokinins, abscisic acid, brassinosterioids, ethylene and salicylic acid. These results demonstrate that AA deficiency modifies hormone signaling in plants, redox-ethylene interactions providing a regulatory node controlling shoot biomass accumulation. Copyright © 2018 Elsevier Inc. All rights reserved.

Content and bioconcentration of mercury in mushrooms from northern Poland.

PubMed

Falandysz, J; Gucia, M; Brzostowski, A; Kawano, M; Bielawski, L; Frankowska, A; Wyrzykowska, B

2003-03-01

Mercury (Hg) was quantified using cold vapour-atomic absorption spectrometry (CV-AAS) in the fruiting bodies of nine edible and five inedible mushrooms and in underlying soil substrate samples. In total, 404 samples comprising caps and stalks and 202 samples of soil substrate (0-10 cm layer) were collected in 1996 from Trójmiejski Landscape Park, northern Poland. Mean Hg concentrations in the soil substrate for different species varied between 10 +/- 3 and 780 +/- 500 ng x g(-1) dry wt (range 2.3-1700). Among edible mushroom species, Horse Mushroom (Agaricus arvensis), Brown Birch Scaber Stalk (Leccinum scabrum), Parasol Mushroom (Macrolepiota procera), King Bolete (Boletus edulis) and Yellow-cracking Bolete (Xerocomus subtomentosus) contained elevated concentrations of Hg ranging from 1600 +/- 930 to 6800 +/- 4000 ng x g(-1) dry wt in the caps. Concentrations of Hg in the stalks were 2.6 +/- 1.1 to 1.7 +/- 1.0 times lower than those in the caps. Some mushroom species investigated had high Hg levels when compared with specimens collected from the background reference sites elsewhere (located far away from the big cities) in northern Poland. Bioconcentration factors of Hg in the caps of Horse Mushroom, Parasol Mushroom and Brown Birch Scaber Stalk were between 150 +/- 58 and 230 +/- 150 ng x g(-1) dry wt, respectively, and for inedible Pestle-shaged Puffball (Claviata excipulformis) was 960 +/- 300 ng x g(-1) dry wt. Linear regression coefficients between Hg in caps and in stalks and Hg soil concentrations showed a positive relationship for A. arvensis and Horse mushroom (p < 0.05) and a negative correlation for the caps of Death Caps (Amanita phalloides) and Woolly Milk Cap (Lactarius torminosus) (p < 0.05), while for other species no clear trend was found.

Accumulation of heavy metals in freshwater fish in cage aquaculture at Cirata Reservoir, West Java, Indonesia.

PubMed

Salami, Indah Rachmatiah Siti; Rahmawati, Suphia; Sutarto, Ratri Indri Hapsari; Jaya, Pinilih Marendra

2008-10-01

Freshwater fish demand in West Java is supported by cage aquaculture in reservoirs. Cirata Reservoir is one of three cascading hydropower reservoirs built along the Citarum River that receives domestic and industrial wastes. Water pollution by heavy metals, such as copper and lead, increases the health risk of humans who consume the fish. The study was aimed to evaluate the concentration of copper and lead in fish, specifically on Cyprinus carpio, that are widely cultivated in the Cirata Reservoir. Two sizes of fish were collected from five sampling points around floating-cage area. The liver of the fish was found to contain the highest copper level, followed by gills, skin, and muscle. In contrast, lead was also high in the liver, followed by skin, gills, and the muscle, but the copper concentration found in the tissues studied was much higher (31.111 +/- 17.911 mg/kg dry wt) than the lead content (0.290 +/- 0.346 mg/kg dry wt). The concentration of metals in smaller fish was always higher than than that in bigger fish. The metals content was also compared to those in fish exposed in a semistatic laboratory-scale study using Oreochromis niloticus. After 28 days exposure with 0.01 mg Cu/L, fish accumulated 21.53 mg Cu/kg dry wt, whereas for Pb exposure of 0.016 mg/L, fish accumulated up to 7 mg/kg dry wt. However, estimates of Cu and Pb intake from C. carpio consumption were still below the average daily intake (ADI) limit. This study suggested that monitoring of water quality and heavy metals in cultured fish is important in protecting human health.

²H enrichment distribution of hepatic glycogen from ²H₂O reveals the contribution of dietary fructose to glycogen synthesis.

PubMed

Delgado, Teresa C; Martins, Fátima O; Carvalho, Filipa; Gonçalves, Ana; Scott, Donald K; O'Doherty, Robert; Macedo, M Paula; Jones, John G

2013-02-15

Dietary fructose can benefit or hinder glycemic control, depending on the quantity consumed, and these contrasting effects are reflected by alterations in postprandial hepatic glycogen synthesis. Recently, we showed that ²H enrichment of glycogen positions 5 and 2 from deuterated water (²H₂O) informs direct and indirect pathway contributions to glycogenesis in naturally feeding rats. Inclusion of position 6(S) ²H enrichment data allows indirect pathway sources to be further resolved into triose phosphate and Krebs cycle precursors. This analysis was applied to six rats that had fed on standard chow (SC) and six rats that had fed on SC plus 35% sucrose in their drinking water (HS). After 2 wk, hepatic glycogenesis sources during overnight feeding were determined by ²H₂O administration and postmortem analysis of glycogen ²H enrichment at the conclusion of the dark period. Net overnight hepatic glycogenesis was similar between SC and HS rodents. Whereas direct pathway contributions were similar (403 ± 71 μmol/g dry wt HS vs. 578 ± 76 μmol/g dry wt SC), triose phosphate contributions were significantly higher for HS compared with SC (382 ± 61 vs. 87 ± 24 μmol/g dry wt, P < 0.01) and Krebs cycle inputs lower for HS compared with SC (110 ± 9 vs. 197 ± 32 μmol/g dry wt, P < 0.05). Analysis of plasma glucose ²H enrichments at the end of the feeding period also revealed a significantly higher fractional contribution of triose phosphate to plasma glucose levels in HS vs. SC. Hence, the ²H enrichment distributions of hepatic glycogen and glucose from ²H₂O inform the contribution of dietary fructose to hepatic glycogen and glucose synthesis.

Impaired growth in the polychaete Armandia brevis exposed to tributyltin in sediment.

PubMed

Meador, J P; Rice, C A

2001-03-01

Juveniles of the opheliid polychaete, Armandia brevis, were exposed to sediment-associated tributyltin (TBT) for 42 days to evaluate toxicity and bioaccumulation. Growth in this species was inhibited in a dose-response fashion by increasing concentrations of TBT. Even though the biota-sediment accumulation factor (BSAF) for TBT declined for the higher sediment concentrations, the total butyltins in tissue increased over all sediment concentrations. At the highest sediment concentrations, polychaetes bioaccumulated less TBT than expected, which was most likely due to reduced uptake and continued metabolism of the parent compound. The less than expected BSAF values exhibited by animals at the exposure concentrations causing severe effects are an important finding for assessing responses in the field. It appears that severe biological effects can occur in long-term experiments without the expected high tissue concentrations; an observation likely explained by altered toxicokinetics. Analysis of variance determined the lowest observed effect concentration for growth to be 191 ng/g sediment dry wt. for 21 days of exposure and 101 ng/g sediment dry wt. at day 42, indicating that 21 days was insufficient for delineating the steady-state toxicity response. When based on regression analysis, the sediment concentration causing a 25% inhibition in growth at 42 days exposure was 93 ng/g dry wt. (total organic carbon = 0.58%). A dose-response association was also determined for polychaete net weight and TBT in tissue. The tissue residue associated with a 25% reduction in growth was 2834 ng/g dry wt. at day 42. A comparison of these results with previous work indicates that juveniles are approximately three times more sensitive than adults to TBT exposure. The sediment concentrations affecting growth in this species are commonly found in urban waterways indicating potentially severe impacts for this and other sensitive species.

The predominant WT1 isoform (+KTS) encodes a DNA-binding protein targeting the planar cell polarity gene Scribble in renal podocytes.

PubMed

Wells, Julie; Rivera, Miguel N; Kim, Woo Jae; Starbuck, Kristen; Haber, Daniel A

2010-07-01

WT1 encodes a tumor suppressor first identified by its inactivation in Wilms' Tumor. Although one WT1 splicing variant encodes a well-characterized zinc finger transcription factor, little is known about the function of the most prevalent WT1 isoform, whose DNA binding domain is disrupted by a three-amino acid (KTS) insertion. Using cells that conditionally express WT1(+KTS), we undertook a genome-wide chromatin immunoprecipitation and cloning analysis to identify candidate WT1(+KTS)-regulated promoters. We identified the planar cell polarity gene Scribble (SCRB) as the first WT1(+KTS) target gene in podocytes of the kidney. WT1 and SCRB expression patterns overlap precisely in developing renal glomeruli of mice, and WT1(+KTS) binds to a 33-nucleotide region within the Scribble promoter in mouse and human cell lines and kidneys. Together, our results support a role for the predominant WT1(+KTS) isoform in transcriptional regulation and suggest a link between the WT1-dependent tumor suppressor pathway and a key component of the planar cell polarity pathway.

Effective Utilization of Carbohydrate in Corncob to Synthesize Furfuralcohol by Chemical-Enzymatic Catalysis in Toluene-Water Media.

PubMed

Xue, Xin-Xia; Di, Jun-Hua; He, Yu-Cai; Wang, Bing-Qian; Ma, Cui-Luan

2018-05-01

In this study, carbohydrates (cellulose plus hemicellulose) in corncob were effectively converted furfuralcohol (FOL) via chemical-enzymatic catalysis in a one-pot manner. After corncob (2.5 g, dry weight) was pretreated with 0.5 wt% oxalic acid, the obtained corncob-derived xylose (19.8 g/L xylose) could be converted to furfural at 60.1% yield with solid acid catalyst SO 4 2- /SnO 2 -attapulgite (3.6 wt% catalyst loading) in the water-toluene (3:1, v/v) at 170 °C for 20 min. Moreover, the oxalic acid-pretreated corncob residue (1.152 g, dry weight) was enzymatically hydrolyzed to 0.902 g glucose and 0.202 g arabinose. Using the corncob-derived glucose (1.0 mM glucose/mM furfural) as cosubstrate, the furfural liquor (48.3 mM furfural) was successfully biotransformed to FOL by recombinant Escherichia coli CCZU-A13 cells harboring an NADH-dependent reductase (SsCR) in the water-toluene (4:1, v/v) under the optimum conditions (50 mM PEG-6000, 0.2 mM Zn 2+ , 0.1 g wet cells/mL, 30 °C, pH 6.5). After the bioreduction for 2 h, FAL was completely converted to FOL. The FOL yield was obtained at 0.11 g FOL/g corncob. Clearly, this one-pot synthesis strategy shows high potential application for the effective synthesis of FOL.

Effect of textile waste water on tomato plant, Lycopersicon esculentum.

PubMed

Marwari, Richa; Khan, T I

2012-09-01

In this study Sanganer town, Jaipur was selected as study area. The plants of Lycopersicon esculentum var. K 21(Tomato) treated with 20 and 30% textile wastewater were analyzed for metal accumulation, growth and biochemical parameters at per, peak and post flowering stages. Findings of the study revealed that chlorophyll content was most severely affected with the increase in metal concentration. Total chlorophyll content showed a reduction of 72.44% while carbohydrate, protein and nitrogen content showed a reduction of 46.83, 71.65 and 71.65% respectively. With the increase in waste water treatment the root and shoot length, root and shoot dry weight and total dry weight were reduced to 50.55, 52.06, 69.93, 72.42, 72.10% respectively. After crop harvesting, the fruit samples of the plants treated with highest concentration of textile waste water contained 2.570 mg g(-1)d.wt. of Zn, 0.800 mg g(-1) d.wt. Cu, 1.520 mg g(-1) d.wt. Cr and 2.010 mg g(-1) d.wt. Pb.

Advances of two-stage riser catalytic cracking of heavy oil for maximizing propylene yield (TMP) process.

PubMed

Chaohe, Yang; Xiaobo, Chen; Jinhong, Zhang; Chunyi, Li; Honghong, Shan

Two-stage riser catalytic cracking of heavy oil for maximizing propylene yield (TMP) process proposed by State Key Laboratory of Heavy oil Processing, China University of Petroleum, can remarkably enhance the propylene yield and minimize the dry gas and coke yields, and obtain high-quality light oils (gasoline and diesel). It has been commercialized since 2006. Up to now, three TMP commercial units have been put into production and other four commercial units are under design and construction. The commercial data showed that taking paraffinic based Daqing (China) atmospheric residue as the feedstock, the propylene yield reached 20.31 wt%, the liquid products yield (the total yield of liquefied petroleum gas, gasoline, and diesel) was 82.66 wt%, and the total yield of dry gas and coke was 14.28 wt%. Moreover, the research octane number of gasoline could be up to 96.

Cell wall properties in Oryza sativa influence mesophyll CO2 conductance.

PubMed

Ellsworth, Patrícia V; Ellsworth, Patrick Z; Koteyeva, Nuria K; Cousins, Asaph B

2018-04-20

Diffusion of CO 2 from the leaf intercellular air space to the site of carboxylation (g m ) is a potential trait for increasing net rates of CO 2 assimilation (A net ), photosynthetic efficiency, and crop productivity. Leaf anatomy plays a key role in this process; however, there are few investigations into how cell wall properties impact g m and A net . Online carbon isotope discrimination was used to determine g m and A net in Oryza sativa wild-type (WT) plants and mutants with disruptions in cell wall mixed-linkage glucan (MLG) production (CslF6 knockouts) under high- and low-light growth conditions. Cell wall thickness (T cw ), surface area of chloroplast exposed to intercellular air spaces (S c ), leaf dry mass per area (LMA), effective porosity, and other leaf anatomical traits were also analyzed. The g m of CslF6 mutants decreased by 83% relative to the WT, with c. 28% of the reduction in g m explained by S c . Although A net /LMA and A net /Chl partially explained differences in A net between genotypes, the change in cell wall properties influenced the diffusivity and availability of CO 2 . The data presented here indicate that the loss of MLG in CslF6 plants had an impact on g m and demonstrate the importance of cell wall effective porosity and liquid path length on g m . © 2018 The Authors. New Phytologist © 2018 New Phytologist Trust.

Ingredient selection for plastic composite supports for L-(+)-lactic acid biofilm fermentation by Lactobacillus casei subsp. rhamnosus.

PubMed Central

Ho, K L; Pometto, A L; Hinz, P N; Dickson, J S; Demirci, A

1997-01-01

Plastic composite supports containing 50% agricultural products (oat hulls, soybean hulls, yeast extract, soybean flour, dried bovine erythrocytes, bovine albumin, and/or mineral salts) and 50% (wt/wt) polypropylene were produced by high-temperature twin-screw extrusion. The research employed two half sets of a five-factorial fractional design (2(5 - 1)) to evaluate the effects of different agricultural components on the properties of the plastic composite supports and to select the best plastic composite support formulation for lactic acid fermentation. The biofilm population was affected by the contact angle and relative hydrophobicity of the supports (r = 0.79 to 0.82). Lactic acid was produced by the suspended cells (r = 0.96) and the biofilm on the plastic composite support discs (r = 0.85). Incorporation of yeast extract into plastic composite supports enhanced growth of free and attached cells in minimal medium (P < 0.0001). The presence of soybean hulls, yeast extract, or mineral salts in plastic composite supports produced less hydrophobic supports (P < 0.0001) and enhanced cell attachment (P < 0.03). Under all conditions, suspended-cell and polypropylene disc controls gave negligible lactic acid production and cell density. Plastic composite supports containing soybean hulls, yeast extract, soybean flour, bovine albumin, and mineral salts gave the highest biofilm population (2.3 x 10(9) CFU/g of support), cell density (absorbance of 1.8 at 620 nm), and lactic acid concentration (7.6 g/liter) in minimal medium. PMID:9212402

Spatial distribution of polycyclic aromatic hydrocarbon and polychlorinated biphenyl sources in the Nakdong River Estuary, South Korea.

PubMed

Lee, Jun H; Woo, Han J; Jeong, Kap S; Kang, Jeong W; Choi, Jae U; Jeong, Eun J; Park, Kap S; Lee, Dong H

2017-10-15

Our research team investigated the elemental composition and the presence of various toxic organic compounds, such as polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs), in estuary surface sediments to trace the spatial distribution of the sources of pollution deposited in Nakdong River, Busan, South Korea. The spatial patterns of elemental composition and toxic organic compounds were determined from the measurements of total organic carbon (TOC), total nitrogen, total sulfur, PAHs, and PCBs. The sediments had TOC contents of between 0.02 and 1.80 wt% (avg. 0.34 wt%), depending on the amount of clay-sized particles. The concentrations of PAHs and PCBs (10.8-167.7 ng g -1 dry wt and 197.0-754.0 pg g -1 dry wt, respectively) in surface sediments revealed different spatial patterns for these compounds, suggesting that they partially originated from the combustion of fossil fuels and from the use of commercial PCB products at adjacent industrial complexes. Although these concentrations were far below the Sediment Quality Guideline (SQG) of the National Oceanic and Atmospheric Administration (NOAA), the sediments at one site contained PCBs at concentrations close to the response level (754.0 pg g -1 dry wt), and were dominated by low-molecular-weight PAHs. The PAHs and PCBs in Nakdong River Estuary sediments were likely to have originated from the combustion of fossil fuels and biomass at the adjacent industrial complexes. The primarily analyzed results determined that PAHs originated from the combustion of fossil fuels and biomass, and overall concentrations were related to the contributions of individual PAHs in most sediment samples. Based on the SQG of the NOAA, our results indicate that the anthropogenic activity should be considered on the future-sustainable management of this estuary system.

Powdery Emulsion Explosive: A New Excellent Industrial Explosive

NASA Astrophysics Data System (ADS)

Ni, Ouqi; Zhang, Kaiming; Yu, Zhengquan; Tang, Shujuan

2012-07-01

Powdery emulsion explosive (PEE), a new powdery industrial explosive with perfect properties, has been made using an emulsification-spray drying technique. PEE is composed of 91-92.5 wt% ammonium nitrate (AN), 4.5-6 wt% organic fuels, and 1.5-1.8 wt% water. Due to its microstructure as a water-in-oil (W/O) emulsion and low water content, it has excellent detonation performance, outstanding water resistance, reliable safety, and good application compared with other industrial explosives, such as ammonite, emulsion explosives, and ANFO.

Preparation of Large-Diameter GaAs Crystals.

DTIC Science & Technology

1981-09-18

ionized impurity content for 40 n-type semi-insulating GaAs. Figure 17 Analysis (in wt %) of impurities in B203 after crystal growth 41 from PBN and quartz...encapsulant to the generation of defect clusters in LEC InP. (15 ) Statistics relative to the incidence of twinning for growth with dry ( ppm wt OH...and wet (> 1000 ppm wt OH) B203 are given in Fig. 5 for growths from fused-SiO 2 and PBN crucibles. A crystal is defined as having twinned if it

DDTs in mothers' milk, placenta and hair, and health risk assessment for infants at two coastal and inland cities in China.

PubMed

Man, Yu Bon; Chan, Janet Kit Yan; Wang, Hong Sheng; Wu, Sheng Chun; Wong, Ming Hung

2014-04-01

This study is a one of the very few investigating the dichloro-diphenyl-trichloroethanes (DDTs) (summation of o,p'-DDE, p,p'-DDE, o,p'-DDD, p,p'-DDD, o,p'-DDT, and p,p'-DDT) in multiple human matrices in mothers' milk, placenta and hair collected from residents from two coastal cities: Guiyu (GY) and Taizhou (TZ) and one inland city: Lin'an (LA). TZ (milk: 360±319ng/g lipid wt.) showed significantly higher concentrations of DDTs than those from LA (milk: 190±131ng/g lipid wt.), whereas, concentrations of DDTs in GY (milk: 305±109ng/g lipid wt.) were in between TZ and LA. In addition, levels of DDTs in the human tissues from TZ (placenta: 122±109ng/g lipid wt.; hair: 79.9±215ng/g dry wt.) were significantly higher than those from Lin'an (placenta: 49.2±30.2ng/g lipid wt.; hair: 10.8±7.09ng/g dry wt.). The above concentrations of DDTs in milk exceeded the Codex Maximum Residue Limits/Extraneous Maximum Residue Limits for milk (20ng/g lipid wt. whole milk), indicating that the human milk samples were grossly polluted. The present study revealed that human specimens collected from the coastal city (TZ) were more contaminated with inland one (LA), based on the levels of DDTs contained in samples which may be due to the higher dietary exposure to DDTs via consumption of contaminated seafood. The estimated daily intakes of DDTs by GY, TZ and LA infants were 1.69±1.86, 1.48±0.79, and 0.95±0.73μg/kg body wt./day, respectively which did not exceed 10μg/kg body wt./day, the provisional tolerable daily intake proposed by the Food and Agriculture Organization/World Health Organization. Copyright © 2014 Elsevier Ltd. All rights reserved.

The characterization of ceramic alumina prepared by using additive glass beads

NASA Astrophysics Data System (ADS)

Suprapedi; Muljadi; Sardjono, Priyo

2018-01-01

The ceramic alumina has been made by using additive glass bead (5 and 10 % wt.). There are two kinds of materials, such as : gamma Alumina and glass bead. Synthesis of alumina was done by ball milling for 24 hours, then the mixed powder was dried in drying oven at 100 °C for 6 hours. Furthermore, the dried powder was mixed by using 2 % of PVA and continued with compacted to form a pellet with pressure of 50 MPA. The next step is sintering process with variation temperature of 1150, 1200, 1250, 1300 and 1400 °C and holding time for 2 hours. The characterization conducted are consist of test density, hardness, shrinkage, and microstructure. The results show that ceramic alumina with addition of 10 % wt. glass bead has the higher value of density, hardness and shrinkage than addition of 5% wt. glass bead. The highest characterization of ceramic alumina with addition 10 % glass bead was achieved at sintering temperature of 1400 °C with density 3.68 g/cm3, hardness vickers 780.40 Hv and shrinkage 15.23 %. The XRD results show that it was founds a corrundum (alpha Alumina) as dominant phase and mullite as minor phase.

Preparation and characteristic of gelatine/oxidized corn starch and gelatin/corn starch blend microspheres.

PubMed

Chen, Hui; Shan, Zhi Hua; Woo, Meng Wai; Chen, Xiao Dong

2017-01-01

Combinations of gelatin (G) and oxidized corn starch (OCS) were explored as a new microcapsule composite for single droplet spray drying. The blending solutions property, gel time, transparency and viscosity of G/CS (corn starch) and G/OCS blend solutions were compared at different ratios (10:0;9:1;8:2;7:3;6:4;5:5) and concentrations(1%wt; 3%wt; 5%wt). The drying and dissolution behaviors of composite droplet have been studied using the single droplet drying technique. Possible reaction mechanisms in the composite blend were elucidated by SEM and FTIR techniques. Blends solutions of G/OCS showed longer Gel time, higher transparency and lower viscosity; further displayed faster dissolution rate than that of G/CS under similar conditions. This was attributed to the formed Schiff base between the aldehyde group of OCS and amino group of G which improved the compatibility between G and OCS. All results indicated that the composites could be prepared with excellent properties by G/OCS (6:4) which would overcome some disadvantage such as thermodynamic incompatibility and phase separation by G/CS. Copyright © 2016. Published by Elsevier B.V.

Metal accumulation in marine bivalves under various tributyltin burdens.

PubMed

Tang, Chuan-Ho; Lin, Chan-Shing; Wang, Wei-Hsien

2009-11-01

In the present study, a field survey was conducted to measure the accumulation of butyltin, Cu, Zn, and Cd in green mussels (Perna viridis) and Pacific oysters (Crassostrea gigas) at the regions along a tributyltin pollution gradient. A negative correlation was found between the tributyltin/total butyltin ratio (0.87-0.31) and tributyltin content (114-5,817 ng/g as tin dry wt) in oysters, while the Cu content (44.2-381 mg/kg dry wt) was positively correlated with the logarithm of tributyltin content during the summer and winter. This suggests that as the tributyltin burden increases, the rates of tributyltin metabolism may be elevated, leading to enhanced Cu accumulation. A similar accumulation pattern for Zn was also found in oysters. In mussels, however, the tributyltin/total butyltin ratio and the Cu and Zn contents remained relatively constant (~ 0.7, 12, and 100 mg/kg dry wt, respectively) regardless of the tributyltin burden. Clearly, the butyltin and Cu/Zn accumulation processes in oysters differ from those in mussels under tributyltin pollution. These observations provide valuable information for those who evaluate or compare tributyltin and/or Cu/Zn pollution using oysters and mussels as bioindicators.

Dry (Mg,Fe)SiO 3 perovskite in the Earth's lower mantle

DOE PAGES

Panero, Wendy R.; Pigott, Jeffrey S.; Reaman, Daniel M.; ...

2015-02-26

Combined synthesis experiments and first-principles calculations show that MgSiO 3-perovskite with minor Al or Fe does not incorporate significant OH under lower mantle conditions. Perovskite, stishovite, and residual melt were synthesized from natural Bamble enstatite samples (Mg/(Fe+Mg) = 0.89 and 0.93; Al 2O 3 < 0.1 wt% with 35 and 2065 ppm wt H 2O, respectively) in the laser-heated diamond anvil cell at 1600-2000 K and 25-65 GPa. Combined Fourier transform infrared (FTIR) spectroscopy, x-ray diffraction, and ex-situ transmission electron microscopy (TEM) analysis demonstrates little difference in the resulting perovskite as a function of initial water content. Four distinct OHmore » vibrational stretching bands are evident upon cooling below 100 K (3576, 3378, 3274, and 3078 cm -1), suggesting 4 potential bonding sites for OH in perovskite with a maximum water content of 220 ppm wt H 2O, and likely no more than 10 ppm wt H 2O. Complementary, Fe-free, first-principles calculations predict multiple potential bonding sites for hydrogen in perovskite, each with significant solution enthalpy (0.2 eV/defect). We calculate that perovskite can dissolve less than 37 ppm wt H 2O (400 ppm H/Si) at the top of the lower mantle, decreasing to 31 ppm wt H 2O (340 ppm H/Si) at 125 GPa and 3000 K in the absence of a melt or fluid phase. Here, we propose that these results resolve a long-standing debate of the perovskite melting curve and explain the order of magnitude increase in viscosity from upper to lower mantle.« less

The predominant WT1 isoform (+KTS) encodes a DNA binding protein targeting the planar cell polarity gene Scribble in renal podocytes

PubMed Central

Wells, Julie; Rivera, Miguel N.; Kim, Woo Jae; Starbuck, Kristen; Haber, Daniel A.

2010-01-01

WT1 encodes a tumor suppressor, first identified by its inactivation in Wilms Tumor. While one WT1 splicing variant encodes a well-characterized zinc finger transcription factor, little is known about the function of the most prevalent WT1 isoform, whose DNA binding domain is disrupted by a three amino acid (KTS) insertion. Using cells which conditionally express WT1(+KTS), we undertook a genome-wide chromatin immunoprecipitation and cloning (ChIP-cloning) analysis to identify candidate WT1(+KTS) regulated promoters. We identified the planar cell polarity (PCP) gene Scribble (SCRB) as the first WT1(+KTS) target gene in podocytes of the kidney. WT1 and SCRB expression patterns overlap precisely in developing renal glomeruli of mice, and WT1(+KTS) binds to a 33 nucleotide region within the Scribble promoter in both mouse and human cell lines and kidneys. Together, our results support a role for the predominant WT1(+KTS) isoform in transcriptional regulation and suggest a link between the WT1-dependent tumor suppressor pathway and a key component of the planar cell polarity pathway. PMID:20571064

Antioxidant, antimutagenic, and antitumor effects of pine needles (Pinus densiflora).

PubMed

Kwak, Chung Shil; Moon, Sung Chae; Lee, Mee Sook

2006-01-01

Pine needles (Pinus densiflora Siebold et Zuccarini) have long been used as a traditional health-promoting medicinal food in Korea. To investigate their potential anticancer effects, antioxidant, antimutagenic, and antitumor activities were assessed in vitro and/or in vivo. Pine needle ethanol extract (PNE) significantly inhibited Fe(2+)-induced lipid peroxidation and scavenged 1,1-diphenyl- 2-picrylhydrazyl radical in vitro. PNE markedly inhibited mutagenicity of 2-anthramine, 2-nitrofluorene, or sodium azide in Salmonella typhimurium TA98 or TA100 in Ames tests. PNE exposure effectively inhibited the growth of cancer cells (MCF-7, SNU-638, and HL-60) compared with normal cell (HDF) in 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. In in vivo antitumor studies, freeze-dried pine needle powder supplemented (5%, wt/wt) diet was fed to mice inoculated with Sarcoma-180 cells or rats treated with mammary carcinogen, 7,12-dimethylbenz[a]anthracene (DMBA, 50 mg/kg body weight). Tumorigenesis was suppressed by pine needle supplementation in the two model systems. Moreover, blood urea nitrogen and aspartate aminotransferase levels were significantly lower in pine needle-supplemented rats in the DMBA-induced mammary tumor model. These results demonstrate that pine needles exhibit strong antioxidant, antimutagenic, and antiproliferative effects on cancer cells and also antitumor effects in vivo and point to their potential usefulness in cancer prevention.

Sodium Chloride Reduces Production of Curvacin A, a Bacteriocin Produced by Lactobacillus curvatus Strain LTH 1174, Originating from Fermented Sausage

PubMed Central

Verluyten, Jurgen; Messens, Winy; De Vuyst, Luc

2004-01-01

Lactobacillus curvatus LTH 1174, a strain originating in fermented sausage, produces the antilisterial bacteriocin curvacin A. Its biokinetics of cell growth and bacteriocin production as a function of various concentrations of salt (sodium chloride) were investigated in vitro during laboratory fermentations using modified MRS medium. A model was set up to describe the effects of different NaCl concentrations on microbial behavior. Both cell growth and bacteriocin activity were affected by changes in the salt concentration. Sodium chloride clearly slowed down the growth of L. curvatus LTH 1174, but more importantly, it had a detrimental effect on specific curvacin A production (kB) and hence on overall bacteriocin activity. Even a low salt concentration (2%, wt/vol) decreased bacteriocin production, while growth was unaffected at this concentration. The inhibitory effect of NaCl was mainly due to its role as an aw-lowering agent. Further, it was clear that salt interfered with bacteriocin induction. Additionally, when 6% (wt/vol) sodium chloride was added, the minimum biomass concentration necessary to start the production of curvacin A (XB) was 0.90 g (cell dry mass) per liter. Addition of the cell-free culture supernatant or a protein solution as a source of induction factor resulted in a decrease in XB, an increase in kB, and hence an increase in the maximum attainable bacteriocin activity. PMID:15066822

Effect of hydrophobic additive on oxygen transport in catalyst layer of proton exchange membrane fuel cells

NASA Astrophysics Data System (ADS)

Wang, Shunzhong; Li, Xiaohui; Wan, Zhaohui; Chen, Yanan; Tan, Jinting; Pan, Mu

2018-03-01

Oxygen transport resistance (OTR) is a critical factor influencing the performance of proton exchange membrane fuel cells (PEMFCs). In this paper, an effective method to reduce the OTR of catalyst layers (CLs) by introducing a hydrophobic additive into traditional CLs is proposed. A low-molecular-weight polytetrafluoroethylene (PTFE) is selected for its feasibility to prepare an emulsion, which is mixed with a traditional catalyst ink to successfully fabricate the CL with PTFE of 10 wt%. The PTFE film exists in the mesopores between the carbon particles. The limiting current of the hydrophobic CL was almost 4000 mA/cm2, which is 500 mA/cm2 higher than that of the traditional CL. PTFE reduces the OTR of the CL in the dry region by as much as 24 s/m compared to the traditional CL and expands the dry region from 2000 mA/cm2 in the traditional CL to 2500 mA/cm2. Furthermore, the CL with the hydrophobic agent can improve the oxygen transport in the wet region (>2000 mA/cm2) more effectively than that in the dry region. All these results indicate that the CL with the hydrophobic agent shows a superior performance in terms of optimizing water management and effectively reduces the OTR in PEMFCs.

Fibrocytes Regulate Wilms’ Tumor 1-Positive Cell Accumulation in Severe Fibrotic Lung Disease

PubMed Central

Sontake, Vishwaraj; Shanmukhappa, Shiva K.; DiPasquale, Betsy A.; Reddy, Geereddy B.; Medvedovic, Mario; Hardie, William D.; White, Eric S.; Madala, Satish K.

2015-01-01

Collagen-producing myofibroblast transdifferentiation is considered a crucial determinant in the formation of scar tissue in the lungs of patients with idiopathic pulmonary fibrosis (IPF). Multiple resident pulmonary cell types and bone marrow-derived fibrocytes have been implicated as contributors to fibrotic lesions due to the transdifferentiation potential of these cells into myofibroblasts. In this study, we assessed the expression of Wilms’ tumor 1 (WT1), a known marker of mesothelial cells, in various cell types in normal and fibrotic lungs. We demonstrate that WT1 is expressed by both mesothelial and mesenchymal cells in IPF lungs, but has limited or no expression in normal human lungs. We also demonstrate that WT1-positive cells accumulate in fibrotic lung lesions, using two different mouse models of pulmonary fibrosis and WT1 promoter-driven fluorescent reporter mice. Reconstitution of bone-marrow cells into a transforming growth factor-α transgenic-mouse model demonstrated that fibrocytes do not transform into WT1-positive mesenchymal cells, but do augment accumulation of WT1-positive cells in severe fibrotic lung disease. Importantly, the number of WT1-positive cells in fibrotic lesions were correlated with severity of lung disease as assessed by changes in lung function, histology, and hydroxyproline levels in mice. Finally, inhibition of WT1 expression was sufficient to attenuate collagen and other extracellular-matrix gene production by mesenchymal cells from both murine and human fibrotic lungs. Thus, the results of this study demonstrate a novel association between fibrocyte-driven WT1-positive cell accumulation and severe fibrotic lung disease. PMID:26371248

Investigations on Local Seismic Phases and Modeling of Seismic Signals

DTIC Science & Technology

1993-10-31

basement is 1 km. The water table, wt , is from Doty and Thordarson (1983). It separates the dry, DT, and the wet, WT, tuff levels. Above these volcanic...regional variations of t*(f) in the United States. Geophys. J. R. astr. Soc. 82 , 125-140 Doty, G. C. and W. Thordarson , 1983. Water table in rocks of

Recovery of Fuel-Precursor Lipids from Oleaginous Yeast

DOE PAGES

Kruger, Jacob S.; Cleveland, Nicholas S.; Yeap, Rou Yi; ...

2018-01-24

Bio-derived lipids offer a potentially promising intermediate to displace petroleum-derived diesel. One of the key challenges for the production of lipids via microbial cell mass is that these products are stored intracellularly and must be extracted and recovered efficiently and economically. Thus, improved methods of cell lysis and lipid extraction are needed. In this study, we examine lipid extraction from wet oleaginous yeast in combination with seven different cell lysis approaches encompassing both physical and chemical techniques (high-pressure homogenization, microwave and conventional thermal treatments, bead beating, acid, base, and enzymatic treatments) to facilitate lipid extraction from a model oleaginous yeastmore » strain, Lipomyces starkeyi. Of the seven techniques investigated, acid treatment led to the highest lipid recovery yields. Further exploration of acid treatment and integration with an economic model revealed that treatment at 170 degrees C for 60 min at 1 wt% H 2SO 4 and 8 wt% yeast solids represents a viable option for both lipid recovery yield and process economics, enabling experimental lipid recovery yields of 88.5-93.0% to be achieved at a corresponding estimated minimum fuel selling price (MFSP) of $5.13-$5.61/gallon of gasoline equivalent (GGE). The same acid treatment conditions applied to two other strains of oleaginous yeast (Cutaneotrichosporon curvatus and Rhodotorula toruloides) resulted in similar lipid recovery yields. In pretreatment experiments scaled up to 300 mL, slightly lower temperatures or shorter pretreatment times, along with higher yeast solids loading, resulted in higher lipid yields than the conditions identified from the small-scale runs. Two replicate runs carried out at 170 degrees C for 30 min using 1 wt% H2SO4 and 19 wt% yeast solids achieved an average lipid recovery of 96.1% at a corresponding estimated MFSP of $4.89/GGE. In all cases, the lipids are primarily triglycerides and free fatty acids comprised mainly of palmitic, stearic, and oleic acids, with smaller fractions of polar lipids. The fatty acid composition of the lipids extracted from the wet treated cell mass is the same as that in freeze-dried whole oleaginous yeast cell mass, suggesting the acid treatment renders all lipids extractable. This work demonstrates that acid treatment is a robust and effective cell lysis technique in a microbial lipid-based biofuel scenario and provides a baseline for further scale-up and process integration.« less

Recovery of Fuel-Precursor Lipids from Oleaginous Yeast

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kruger, Jacob S.; Cleveland, Nicholas S.; Yeap, Rou Yi

Bio-derived lipids offer a potentially promising intermediate to displace petroleum-derived diesel. One of the key challenges for the production of lipids via microbial cell mass is that these products are stored intracellularly and must be extracted and recovered efficiently and economically. Thus, improved methods of cell lysis and lipid extraction are needed. In this study, we examine lipid extraction from wet oleaginous yeast in combination with seven different cell lysis approaches encompassing both physical and chemical techniques (high-pressure homogenization, microwave and conventional thermal treatments, bead beating, acid, base, and enzymatic treatments) to facilitate lipid extraction from a model oleaginous yeastmore » strain, Lipomyces starkeyi. Of the seven techniques investigated, acid treatment led to the highest lipid recovery yields. Further exploration of acid treatment and integration with an economic model revealed that treatment at 170 degrees C for 60 min at 1 wt% H 2SO 4 and 8 wt% yeast solids represents a viable option for both lipid recovery yield and process economics, enabling experimental lipid recovery yields of 88.5-93.0% to be achieved at a corresponding estimated minimum fuel selling price (MFSP) of $5.13-$5.61/gallon of gasoline equivalent (GGE). The same acid treatment conditions applied to two other strains of oleaginous yeast (Cutaneotrichosporon curvatus and Rhodotorula toruloides) resulted in similar lipid recovery yields. In pretreatment experiments scaled up to 300 mL, slightly lower temperatures or shorter pretreatment times, along with higher yeast solids loading, resulted in higher lipid yields than the conditions identified from the small-scale runs. Two replicate runs carried out at 170 degrees C for 30 min using 1 wt% H2SO4 and 19 wt% yeast solids achieved an average lipid recovery of 96.1% at a corresponding estimated MFSP of $4.89/GGE. In all cases, the lipids are primarily triglycerides and free fatty acids comprised mainly of palmitic, stearic, and oleic acids, with smaller fractions of polar lipids. The fatty acid composition of the lipids extracted from the wet treated cell mass is the same as that in freeze-dried whole oleaginous yeast cell mass, suggesting the acid treatment renders all lipids extractable. This work demonstrates that acid treatment is a robust and effective cell lysis technique in a microbial lipid-based biofuel scenario and provides a baseline for further scale-up and process integration.« less

Analysis of the main components of the aguamiel produced by the maguey-pulquero (Agave mapisaga) throughout the harvest period.

PubMed

Ortiz-Basurto, Rosa Isela; Pourcelly, Gérald; Doco, Thierry; Williams, Pascale; Dornier, Manuel; Belleville, Marie-Pierre

2008-05-28

The main characteristics of the aguamiel (maguey-pulquero sap) during the harvest period of the Agave mapisaga plants were assessed to establish its stability through time and the industrial potential of its components. Only minor differences in aguamiel composition were detected among samples collected at different time points of the harvest period. The aguamiel analyzed contained 11.5 wt % of dry matter, which was composed mainly of sugars (75 wt %). Among these sugars, 10 wt % were fructo-oligosaccharides (FOS), which are known to be important in the food industry for their prebiotic properties. Other components include 0.3 wt % of free amino acids (with most essential amino acids and four neurotransmitters: GABA, GLY, GLX, and ASX), 3 wt % of proteins, and 3 wt % of ashes.

Wilms Tumor 1b defines a wound-specific sheath cell subpopulation associated with notochord repair

PubMed Central

Lopez-Baez, Juan Carlos; Zeng, Zhiqiang; Brunsdon, Hannah; Salzano, Angela; Brombin, Alessandro; Wyatt, Cameron; Rybski, Witold; Huitema, Leonie F A; Dale, Rodney M; Kawakami, Koichi; Englert, Christoph; Chandra, Tamir; Schulte-Merker, Stefan

2018-01-01

Regenerative therapy for degenerative spine disorders requires the identification of cells that can slow down and possibly reverse degenerative processes. Here, we identify an unanticipated wound-specific notochord sheath cell subpopulation that expresses Wilms Tumor (WT) 1b following injury in zebrafish. We show that localized damage leads to Wt1b expression in sheath cells, and that wt1b+cells migrate into the wound to form a stopper-like structure, likely to maintain structural integrity. Wt1b+sheath cells are distinct in expressing cartilage and vacuolar genes, and in repressing a Wt1b-p53 transcriptional programme. At the wound, wt1b+and entpd5+ cells constitute separate, tightly-associated subpopulations. Surprisingly, wt1b expression at the site of injury is maintained even into adult stages in developing vertebrae, which form in an untypical manner via a cartilage intermediate. Given that notochord cells are retained in adult intervertebral discs, the identification of novel subpopulations may have important implications for regenerative spine disorder treatments. PMID:29405914

High titer ethanol production from SPORL-pretreated lodgepole pine by simultaneous enzymatic saccharification and combined fermentation

USDA-ARS?s Scientific Manuscript database

Lodgepole wood chips were pretreated by SPORL at 25% solids loading and 180ºC for 20 min with sulfuric acid and sodium bisulfite charges of 2.2 and 8 wt/wt % on oven dry wood basis, respectively. The pretreated wood chips were disk milled with the pretreatment spent liquor and water and then separa...

Potential role of alpha-synuclein and metallothionein in lead-induced inclusion body formation.

PubMed

Zuo, Peijun; Qu, Wei; Cooper, Ryan N; Goyer, Robert A; Diwan, Bhalchandra A; Waalkes, Michael P

2009-09-01

Lead (Pb) produces aggresome-like inclusion bodies (IBs) in target cells as a toxic response. Our prior work shows metallothionein (MT) is required for this process. We used MT-I/II double knockout (MT-null) and parental wild-type (WT) cell lines to further explore the formation process of Pb-induced IBs. Unlike WT cells, MT-null cells did not form IBs after Pb exposure. Western blot of cytosol showed soluble MT protein in WT cells was lost during Pb exposure as IBs formed. Transfection of MT-I into MT-null cells allowed IBs formation after Pb exposure. Considering Pb-induced IBs may be like disease-related aggresomes, which often contain alpha-synuclein (Scna), we investigated Scna expression in cells capable (WT) and incapable (MT-null) of producing IBs after Pb exposure. Scna protein showed poor basal expression in MT-null cells. Pb exposure increased Scna expression only in WT cells. MT transfection increased Scna transcript to WT levels. In WT or MT-transfected MT-null cells, Pb-induced Scna expression rapidly increased and then decreased over 48 h as Pb-induced IBs were formed. A direct interaction between Scna and MT was confirmed ex vivo by antibody pulldown assay where the proteins coprecipitated with an antibody to MT. Pb exposure caused increased colocalization of MT and Scna proteins with time only in WT cells. In WT mice after chronic Pb exposure Scna was localized in renal cells containing forming IBs, whereas MT-null mice did not form IBs. Thus, Scna could be component of Pb-induced IBs and, with MT, may play a role in IBs formation.

Hormone treatment enhances WT1 activation of Renilla luciferase constructs in LNCaP cells.

PubMed

Hanson, Julie; Reese, Jennifer; Gorman, Jacquelyn; Cash, Jennifer; Fraizer, Gail

2007-01-01

The zinc finger transcription factor, WT1, regulates many growth control genes, repressing or activating transcription depending on the gene and cell type. Based on earlier analyses of the effect of WT1 on androgen responsive genes, we hypothesized that there may be an interaction between the androgen signaling pathway and WT1, such that the commonly used Renilla luciferase control vectors were activated in LNCaP prostate cancer cells. Using cotransfection assays we tested the effects of WT1 and/or the androgen analog, R1881, on two Renilla luciferase vectors, pRL-SV40 and the promoter-less pRL-null. To determine whether the zinc finger DNA binding domain was required, the zinc finger mutant DDS-WT1 (R394W) was tested; but it had no significant effect on the Renilla luciferase vectors. To determine whether the androgen signaling pathway was required, WT1 was co-transfected with Renilla vectors in cells with varied hormone responsiveness. The WT1 effect on pRL-null varied from no significant effect in 293 and PC3 cells to very strong enhancement in LNCaP cells treated with 5 nM R1881. Overall, these results suggest that hormone enhanced WT1 mediated activation of Renilla luciferase and that these interactions require an intact WT1 zinc finger DNA binding domain.

Invasive crayfish as vectors of mercury in freshwater food webs of the Pacific Northwest

USGS Publications Warehouse

Johnson, Branden L.; Willacker, James J.; Eagles-Smith, Collin A.; Pearl, Christopher A.; Adams, Michael J.

2014-01-01

Invasive species are important drivers of environmental change in aquatic ecosystems and can alter habitat characteristics, community composition, and ecosystem energetics. Such changes have important implications for many ecosystem processes, including the bioaccumulation and biomagnification of contaminants through food webs. Mercury concentrations were measured in 2 nonnative and 1 native crayfish species from western Oregon (USA). Nonnative red swamp crayfish had mercury concentrations similar to those in native signal crayfish (0.29 ± 0.05 µg/g dry wt and 0.36 ± 0.06 µg/g dry wt, respectively), whereas the nonnative ringed crayfish had lower mercury concentrations (0.10 ± 0.02 µg/g dry wt) than either of the other species. The mean energy content of muscle was similar between the native signal crayfish and nonnative ringed crayfish but was significantly higher in the nonnative red swamp crayfish. Across species, mercury concentrations were negatively correlated with energy density. Such energetic differences could exacerbate changes in mercury transfer through trophic pathways of food webs, especially via alterations to the growth dynamics of consumers. Thus, it is important to consider the role of energy content in determining effective mercury exposure even when mercury concentrations on a per-unit mass basis do not differ between species.

Zinc accumulation by the slime mold Fuligo septica (L.) Wiggers in the former Soviet Union and North Korea.

PubMed

Zhulidov, Daniel A; Robarts, Richard D; Zhulidov, Alexander V; Zhulidova, Olga V; Markelov, Danila A; Rusanov, Viktor A; Headley, John V

2002-01-01

Samples of the slime mold Fuligo septica (L.) Wiggers were collected from an ecologically diverse selection of sites across the former USSR and in North Korea to determine their Zn concentrations. Plasmodia were collected from trees, rocks, soils, the walls of buildings and a variety of other materials and structures from 1990 to 1996. The biomass collected ranged from 305 to 968 mg, whereas Zn concentrations in plasmodia of F. septica ranged from 8400 to 23,000 mg kg(-1) dry wt. (mean and standard error = 14,200 +/- 860 mg kg(-1) dry wt.). No clear trend as to which areas produced F. septica with the highest Zn concentrations was discernable. Nor was it possible to identify any particular substrate on which F. septica grew that produced noticeably high Zn concentrations. For example, forest litter on which F. septica was found had Zn concentrations of only 25 to 130 mg kg(-1) dry wt. Our data confirm the only other study showing hyperaccumulation of Zn in F. septica, which was carried out in Finland. This ability seems to be unique to this species, but how or why it does this, or why such high Zn concentrations are not toxic to F. septica, are questions requiring future research.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Svensson, Emelie; Eriksson, Helena; Gekas, Christos

The Wilms tumor gene 1 (WT1) encodes a zinc-finger-containing transcription factor highly expressed in immature hematopoietic progenitor cells. Overexpression and presence of somatic mutations in acute leukemia indicate a role for WT1 in the pathogenesis of leukemia. CD34{sup +} progenitor cells were transduced with one splice variant of human WT1 without the KTS insert in the zinc-finger domain, WT1(+/-), and with a deleted mutant of WT1 lacking the entire zinc-finger region, WT1(delZ), thus incapable of binding DNA. We show that inhibition of erythroid colony formation and differentiation is absolutely dependent on the DNA-binding zinc-finger domain of WT1. Unexpectedly, however, WT1(delZ)more » was equally effective as wild type protein in the reduction of myeloid clonogenic growth as well as in stimulation of myeloid differentiation, as judged by the expression of cell surface CD11b. Expression of neither WT1(+/-) nor WT1(delZ) upregulated mRNA for the cdk inhibitor p21{sup Waf1/Cip1} or p27{sup Kip1}. Our results demonstrate that WT1 affects proliferation and differentiation in erythroid and myeloid cells by different molecular mechanisms, and suggest that mutations affecting the zinc-finger domain of WT1 could interfere with normal differentiation in the pathogenesis of leukemia.« less

Detection of Wilms' tumor antigen--specific CTL in tumor-draining lymph nodes of patients with early breast cancer.

PubMed

Gillmore, Roopinder; Xue, Shao-An; Holler, Angelika; Kaeda, Jaspal; Hadjiminas, Dimitri; Healy, Vourneen; Dina, Roberto; Parry, Suzanne C; Bellantuono, Ilaria; Ghani, Yasmeen; Coombes, R Charles; Waxman, Jonathan; Stauss, Hans J

2006-01-01

The Wilms' tumor antigen (WT1) is overexpressed in approximately 90% of breast tumors and, thus, is a potential target antigen for the immunotherapy of breast cancer. We have tested the working hypotheses that WT1 can be immunogenic in patients with breast cancer and can stimulate CTL of sufficient avidity to kill tumor cells. Paired tumor-draining lymph node and peripheral blood samples were analyzed from five HLA-A2-positive patients with stage I/II breast cancer. Fluorescent HLA-A*0201/WT1 tetramers were used to quantify WT1-specific CTL and the functional capacity of the CTL was assessed using cytotoxicity assays and intracellular cytokine staining. WT1 tetramer-binding T cells expanded from all lymph node samples but none of the corresponding peripheral blood samples. Functional assays were carried out on T cells from the patient who had yielded the highest frequency of HLA-A*0201/WT1 tetramer-positive cells. The cytotoxicity assays showed WT1 peptide--specific killing activity of the CTL, whereas intracellular cytokine staining confirmed that the tetramer--positive T cells produced IFN-gamma after stimulation with WT1 peptide. These WT1-specific T cells killed HLA-A2-positive breast cancer cell lines treated with IFN-gamma but no killing was observed with untreated tumor cells. These results show that WT1-specific CTL can be expanded from the tumor-draining lymph nodes of breast cancer patients and that they can display peptide-specific effector function. However, the CTL only killed IFN-gamma-treated tumor targets expressing high levels of HLA-A2 and not tumor cells with low HLA expression. This suggests that induction of autologous WT1-specific CTL may offer only limited tumor protection and that strategies that allow a high level of peptide/MHC complex presentation and/or improve CTL avidity may be required.

Co-Introduced Functional CCR2 Potentiates In Vivo Anti-Lung Cancer Functionality Mediated by T Cells Double Gene-Modified to Express WT1-Specific T-Cell Receptor

PubMed Central

Asai, Hiroaki; Fujiwara, Hiroshi; An, Jun; Ochi, Toshiki; Miyazaki, Yukihiro; Nagai, Kozo; Okamoto, Sachiko; Mineno, Junichi; Kuzushima, Kiyotaka; Shiku, Hiroshi; Inoue, Hirofumi; Yasukawa, Masaki

2013-01-01

Background and Purpose Although gene-modification of T cells to express tumor-related antigen-specific T-cell receptor (TCR) or chimeric antigen receptor (CAR) has clinically proved promise, there still remains room to improve the clinical efficacy of re-directed T-cell based antitumor adoptive therapy. In order to achieve more objective clinical responses using ex vivo-expanded tumor-responsive T cells, the infused T cells need to show adequate localized infiltration into the tumor. Methodology/Principal Findings Human lung cancer cells variously express a tumor antigen, Wilms' Tumor gene product 1 (WT1), and an inflammatory chemokine, CCL2. However, CCR2, the relevant receptor for CCL2, is rarely expressed on activated T-lymphocytes. A HLA-A2402+ human lung cancer cell line, LK79, which expresses high amounts of both CCL2 and WT1 mRNA, was employed as a target. Normal CD8+ T cells were retrovirally gene-modified to express both CCR2 and HLA-A*2402-restricted and WT1235–243 nonapeptide-specific TCR as an effector. Anti-tumor functionality mediated by these effector cells against LK79 cells was assessed both in vitro and in vivo. Finally the impact of CCL2 on WT1 epitope-responsive TCR signaling mediated by the effector cells was studied. Introduced CCR2 was functionally validated using gene-modified Jurkat cells and human CD3+ T cells both in vitro and in vivo. Double gene-modified CD3+ T cells successfully demonstrated both CCL2-tropic tumor trafficking and cytocidal reactivity against LK79 cells in vitro and in vivo. CCL2 augmented the WT1 epitope-responsive TCR signaling shown by relevant luciferase production in double gene-modified Jurkat/MA cells to express luciferase and WT1-specific TCR, and CCL2 also dose-dependently augmented WT1 epitope-responsive IFN-γ production and CD107a expression mediated by these double gene-modifiedCD3+ T cells. Conclusion/Significance Introduction of the CCL2/CCR2 axis successfully potentiated in vivo anti-lung cancer reactivity mediated by CD8+ T cells double gene-modified to express WT1-specific TCR and CCR2 not only via CCL2-tropic tumor trafficking, but also CCL2-enhanced WT1-responsiveness. PMID:23441216

A novel approach for improving the drying behavior of sludge by the appropriate foaming pretreatment.

PubMed

Huang, Jing; Yang, Zhao-Hui; Zeng, Guang-Ming; Wang, Hui-Ling; Yan, Jing-Wu; Xu, Hai-Yin; Gou, Cheng-Liu

2015-01-01

Foaming pretreatment has long been recognized to promote drying materials with sticky and viscous behaviors. A novel approach, CaO addition followed by appropriate mechanical whipping, was employed for the foaming of dewatered sludge at a moisture content of 80-85%. In the convective drying, the foamed sludge at 0.70 g/mL had the best drying performance at any given temperature, which saved 35-41% drying time for reaching 20% moisture content compared with the non-foamed sludge. Considering the maximum foaming efficiency, the optimal CaO addition was found at 2.0 wt%. For a better understanding of the foaming mechanisms, the foamability of sludge processed with other pretreatment methods, including NaOH addition (0-3.0 wt%) and heating application (60-120 °C), were investigated while continuously whipping. Their recovered supernatant phases were characterized by pH, surface tension, soluble chemical oxygen demand (sCOD), protein concentration, polysaccharide concentration and spectra of excitation-emission matrices (EEM). These comparative studies indicated that the sludge foaming was mainly derived from the decreased surface tension by the surfactants and the promoted foam persistence by the protein derived compounds. Further, a comprehensive analysis of the sludge drying characteristics was performed including the surface moisture evaporation, the effective moisture diffusivity and the micromorphology of dried sludge. The results indicated that the drying advantages of foamed sludge were mainly attributed to the larger evaporation surface in a limited drying area and the more active moisture capillary movement through the liquid films, which resulted in longer constant evaporation rate periods and better effective moisture diffusivity, respectively.

Biomass production and nutrient removal potential of water hyacinth cultured in sewage effluent

DOE Office of Scientific and Technical Information (OSTI.GOV)

Reddy, K.R.; Hueston, F.M.; McKinn, T.

1985-05-01

Growth and nutrient uptake of water hyacinth (Eichhornia crassipes (Mart Solms)) cultured in sewage effluent were measured over a period of one year in a prototype wastewater treatment system which has been in operation at Walt Disney World near Orlando, Florida. Annual productivity of water hyacinth cultured in primary sewage effluent (Channel II) was found to be in the range of 5 to 27 g dry wt/m/sup 2/ day (23.6 dry tons/acre yr). Average growth rate during the months of May through October 1982 for hyacinth cultured in Channel II (primary sewage effluent) and Channel I (treated primary sewage effluentmore » leaving Channel II) was about 16 g dry wt/m/sup 2/ (27 dry tons/acre yr), compared to the growth rate of 13 g dry wt/m/sup 2/ (22 dry tons/acre yr) for hyacinths cultured in secondary sewage effluent. Plants cultured in secondary sewage effluent generally had longer roots than the plants cultured in primary sewage effluent. A significant relationship was observed between the growth rate of hyacinth and the solar radiation. N and P concentration of the plant tissue were higher in the hyacinths cultured during winter months compared to the plants grown in summer months. Average N and P concentration of the plants cultured im primary sewage effluent were found to be 3.7% N and 0.94% P, respectively, while the plants cultured in secondary sewage effluent had a total N and P content of 2.8% N and 0.79% P. Nutrient ratios of the major plant nurtrients were found to be approximately the same as the nutrient ratios in the sewage effluent. Annual N and P uptake rates of hyacinth cultured in sewage effluent were found to be in the range of 1176 to 1193 kg N/ha yr and 321 to 387 kg P/ha yr, respectively.« less

Renal protection from ischemia mediated by A2A adenosine receptors on bone marrow–derived cells

PubMed Central

Day, Yuan-Ji; Huang, Liping; McDuffie, Marcia J.; Rosin, Diane L.; Ye, Hong; Chen, Jiang-Fan; Schwarzschild, Michael A.; Fink, J. Stephen; Linden, Joel; Okusa, Mark D.

2003-01-01

Activation of A2A adenosine receptors (A2ARs) protects kidneys from ischemia-reperfusion injury (IRI). A2ARs are expressed on bone marrow–derived (BM-derived) cells and renal smooth muscle, epithelial, and endothelial cells. To measure the contribution of A2ARs on BM-derived cells in suppressing renal IRI, we examined the effects of a selective agonist of A2ARs, ATL146e, in chimeric mice in which BM was ablated by lethal radiation and reconstituted with donor BM cells derived from GFP, A2AR-KO, or WT mice to produce GFP→WT, A2A-KO→WT, or WT→WT mouse chimera. We found little or no repopulation of renal vascular endothelial cells by donor BM with or without renal IRI. ATL146e had no effect on IRI in A2A-KO mice or A2A-KO→WT chimera, but reduced the rise in plasma creatinine from IRI by 75% in WT mice and by 60% in WT→WT chimera. ATL146e reduced the induction of IL-6, IL-1β, IL-1ra, and TGF-α mRNA in WT→WT mice but not in A2A-KO→WT mice. Plasma creatinine was significantly greater in A2A-KO than in WT mice after IRI, suggesting some renal protection by endogenous adenosine. We conclude that protection from renal IRI by A2AR agonists or endogenous adenosine requires activation of receptors expressed on BM-derived cells. PMID:12975473

Influence of drying restraint on physical and mechanical properties of nanofibrillated cellulose films

Treesearch

Carlos Baez; John Considine; Robert Rowlands

2014-01-01

Nanofibrillated cellulose (NFC) is a renewable and biodegradable fibril that possesses high strength and stiffness resulting from high level hydrogen bonding. Films made from NFC shrink and distort as they transition from a wet state (20 wt% solids) to a state of moisture equilibrium (90 wt% solids at 50 % RH, 23 °C). Material distortions are driven by development of...

Warthin tumor with signet-ring cell features as a pitfall in salivary gland cytopathology.

PubMed

Bellevicine, Claudio; Iaccarino, Antonino; Malapelle, Umberto; Troncone, Giancarlo

2013-01-01

Warthin tumor (WT) is a common parotid lesion reliably diagnosed by fine-needle aspiration (FNA). Worrisome metaplastic changes may occur in WT. Their interpretation as mucoepidermoid carcinoma represents a diagnostic pitfall. Moreover, WT and mucoepidermoid carcinoma may coexist, making this distinction difficult. So it is worthwhile to report unusual WT features. We describe a WT with signet-ring cells (SRCs). A 61-year-old male presenting with a 3.6-cm right parotid gland mass underwent FNA with rapid on-site evaluation. DiffQuik-stained smears showed groups of oncocytic cells with abundant granular cytoplasm in a background rich with debris, foamy macrophages and lymphoid cells. SRCs were observed on Papanicolaou-stained smears prepared from a second pass. A WT with SRC features was diagnosed. Histology revealed a WT with post-FNA infarctual changes. To avoid false-positive diagnoses, the cytopathologist should be aware that SRC features may occur in WT. The concomitant presence of oncocytes and SRCs is useful to correctly diagnose this unusual WT variant. Copyright © 2013 S. Karger AG, Basel.

Chemo-enzymatic synthesis of furfuralcohol from chestnut shell hydrolysate by a sequential acid-catalyzed dehydration under microwave and Escherichia coli CCZU-Y10 whole-cells conversion.

PubMed

Di, Junhua; Ma, Cuiluan; Qian, Jianghao; Liao, Xiaolong; Peng, Bo; He, Yucai

2018-08-01

In this study, chemo-enzymatic synthesis of furfuralcohol from biomass-derived xylose was successfully demonstrated by a sequential acid-catalyzed dehydration under microwave and whole-cells reduction. After dry dewaxed chestnut shells (CNS, 75 g/L) was acid-hydrolyzed with dilute oxalic acid (0.5 wt%) at 140 °C for 40 min, the obtained CNS-derived xylose (17.9 g/L xylose) could be converted to furfural at 78.8% yield with solid acid SO 4 2- /SnO 2 -Attapulgite (2.0 wt% catalyst loading) in the dibutyl phthalate-water (1:1, v:v) under microwave (600 W) at 180 °C for 10 min. In the dibutyl phthalate-water (1:1, v/v) media at 30 °C and pH 6.5, the furfural liquor (47.0 mM furfural) was biologically converted to furfuralcohol by recombinant Escherichia coli CCZU-Y10 whole-cells harboring an NADH-dependent reductase (PgCR) without extra addition of NAD + and glucose, and furfural was completely converted to furfuralcohol after 2.5 h. Clearly, this one-pot synthesis strategy can be effectively used for furfuralcohol production. Copyright © 2018 Elsevier Ltd. All rights reserved.

Seasonal Variation in Fluoride Content in Groundwaters of Langtang Area, Northcentral Nigeria

NASA Astrophysics Data System (ADS)

Dibal, H. U.; Dajilak, W. N.; Lekmang, I. C.; Nimze, L. W.; Yenne, E. Y.

2017-06-01

Thirty groundwater samples were collected at the peak of the rainy season and analysed for fluoride and other cations and anions in drinking water sources of Langtang area. For comparative purposes, thirty seven groundwater samples were collected in the dry season. The aim of the study was to determine variation in fluoride content with respect to the seasons. Fluoride in water was determined by the Ion Selective Electrode (ISE) and the cations by the Inductively Coupled Plasma Atomic Emission Spectrometry (ICP-AES). The anion (sulphate) was determined by Multi - Ion Colorimeter, bicarbonate and chloride by titration method. In addition fluorine content in aquifer materials from a borehole section were determined by Fusion method. The two seasons show variation in content of fluoride in groundwater. Fluoride content in groundwater is higher in the dry season ranging from 0.13 - 10.3 mg/l compared to the 0.06 - 4.60 mg/l values in the rainy season. Content of fluorine (0.01 wt %) in the aquifer materials (sands) is low from depth of 0 to 7.95 m. However, fluorine content increases with depth, from 7.95 to 10.60 m with concentration of 0.04 wt %, 0.05 wt % from 10.60 to 13.25m, and 0.07 wt % from 13.25 to 15.70 m, the content of fluorine however, decreased at depth 15.70 to18.55m with concentration of 0.02 wt % even with fluorite mineral in the aquifer material at this depth. Dilution of fluoride ion as a result of rain input which recharges the aquifer may be the main reason for lower values recorded in the rainy season. Over fifty and sixty percent of waters in both dry and rainy season have fluoride concentration above the WHO upper limit of 1.5 mg/l. Consumption of these elevated values of fluoride in groundwater of the study area, clearly manifests as symptoms of dental fluorosis.

Regulation of lithospermic acid B and shikonin production in Lithospermum erythrorhizon cell suspension cultures.

PubMed

Yamamoto, Hirobumi; Zhao, Ping; Yazaki, Kazufumi; Inoue, Kenichiro

2002-08-01

Cell suspension cultures of Lithospermum erythrorhizon produced a large amount of lithospermic acid B, a caffeic acid tetramer, as well as shikonin derivatives (each ca. 10% of dry wt.) when cultured in shikonin production medium M-9. Various culture factors for increasing the production of lithospermic acid B were investigated. Lithospermic acid B production was inhibited by 2, 4-D or NH4+, whereas it was stimulated by Cu2+. These regulatory patterns were similar to those for the production of shikonin derivatives in these cell cultures, suggestive of close relations and similar metabolic regulation between the production of these compounds. Cultivation under light illumination, however, showed that these metabolisms were independently regulated. In particular, blue light showed a stimulatory effect on lithospermic acid B production, while shikonin production was strongly inhibited, indicative of an effective condition for lithospermic acid B production.

Transcriptional regulation of human retinoic acid receptor-alpha (RAR-{alpha}) by Wilms` tumour gene product

DOE Office of Scientific and Technical Information (OSTI.GOV)

Goodyer, P.R.; Torban, E.; Dehbi, M.

1994-09-01

The Wilms` tumor gene encodes a 47-49 kDa transcription factor expressed in kidney, gonads and mesothelium during embryogenesis. Inherited mutations of WT1 lead to aberrant urogenital development and Wilms` tumor, but the role of WT1 in development is not fully understood. Since the human RAR-{alpha} gene contains a potential WT1 binding site at its 5{prime} end, we studied the effect of WT1 co-transfection on expression of an RAR-{alpha} promoter/CAT reporter construct in COS cells. COS cells were plated at 5X10{sup 5} cells/dish in DMEM with 10% FBS and transfected by the Ca/PO4 method with an expression plasmid containing the full-lengthmore » WT1 (-/-) cDNA under the control of the CMV promoter, plasmid containing the RAR-{alpha} promoter (-519 to +36)/CAT reporter and TK/growth hormone plasmid to control for efficiency of transfection. CAT/GH activity at 48 hours was inhibited by co-transfection with increasing amounts of WT1 (-/-); maximum inhibition = 5% of control. WT1 co-transfection did not affect expression of TKGH, nor of a CMV-CAT vector. Expression of WT1 protein in tranfected COS cells was demonstrated by Western blotting. Minimal inhibiton of RAR-{alpha}/CAT activity was seen when cells were co-transfected with vectors containing WT1 deletion mutants, alternate WT1 splicing variants, or WT1 (-/-) cDNA bearing a mutation identified in a patient with Drash syndrome. Gel shift assays indicated binding of WT1 to RAR-{alpha} cDNA but not to an RAR-{alpha} deletion mutant lacking the GCGGGGGGCG site. These observations suggest that WT1 may function to regulate RAR-{alpha} expression during normal development.« less

Effect of microwave drying and oven drying on the water activity, color, phenolic compounds content and antioxidant activity of coconut husk (Cocos nucifera L.).

PubMed

Valadez-Carmona, Lourdes; Cortez-García, Rosa María; Plazola-Jacinto, Carla Patricia; Necoechea-Mondragón, Hugo; Ortiz-Moreno, Alicia

2016-09-01

The coconut ( Cocos nucifera L.) husk is basically composed by fiber and pith material and remained under-utilized. This is an important source of phenolic compounds that could be used as functional ingredients. The aim of this study was to determine the effect of: oven-drying (OD) and microwave drying (MD), on the water activity, color, phenolic compound content and antioxidant activity of coconut husk. The OD was performed at 60 °C for 12 h and MD was performed at 900 W for 10 min. The total phenolic content (TPC) in fresh coconut husk was 64.2 mg GAE/g dry wt and significant higher than observed after OD and MD of 35.8 and 45.5 mg GAE/g dry wt, respectively. Ten phenols were identified in fresh and dehydrated coconut husks. The husk MD showed an increase in the content of gallic, 4-hydroxybenzoic, ferulic and syringic acids and epicatechin compared with the fresh; while coconut husk OD and MD, showed a decrease in the content of vanillic acid, vanillin, catequin and kaempferol. The antioxidant activity decreased after both OD and MD. However, MD resulted in a better antioxidant activity in husk than OD. MD of husk resulted into better retention of preserved color, TPC and TFC than OD.

Metallothionein blocks oxidative DNA damage induced by acute inorganic arsenic exposure

DOE Office of Scientific and Technical Information (OSTI.GOV)

Qu, Wei, E-mail: qu@niehs.nih.gov; Waalkes, Michael P.

We studied how protein metallothionein (MT) impacts arsenic-induced oxidative DNA damage (ODD) using cells that poorly express MT (MT-I/II double knockout embryonic cells; called MT-null cells) and wild-type (WT) MT competent cells. Arsenic (as NaAsO{sub 2}) was less cytolethal over 24 h in WT cells (LC{sub 50} = 11.0 ± 1.3 μM; mean ± SEM) than in MT-null cells (LC{sub 50} = 5.6 ± 1.2 μM). ODD was measured by the immuno-spin trapping method. Arsenic (1 or 5 μM; 24 h) induced much less ODD in WT cells (121% and 141% of control, respectively) than in MT-null cells (202% andmore » 260%). In WT cells arsenic caused concentration-dependent increases in MT expression (transcript and protein), and in the metal-responsive transcription factor-1 (MTF-1), which is required to induce the MT gene. In contrast, basal MT levels were not detectable in MT-null cells and unaltered by arsenic exposure. Transfection of MT-I gene into the MT-null cells markedly reduced arsenic-induced ODD levels. The transport genes, Abcc1 and Abcc2 were increased by arsenic in WT cells but either showed no or very limited increases in MT-null cells. Arsenic caused increases in oxidant stress defense genes HO-1 and GSTα2 in both WT and MT-null cells, but to much higher levels in WT cells. WT cells appear more adept at activating metal transport systems and oxidant response genes, although the role of MT in these responses is unclear. Overall, MT protects against arsenic-induced ODD in MT competent cells by potential sequestration of scavenging oxidant radicals and/or arsenic. - Highlights: • Metallothionein blocks arsenic toxicity. • Metallothionein reduces arsenic-induced DNA damage. • Metallothionein may bind arsenic or radicals produced by arsenic.« less

Wilms' tumor blastemal stem cells dedifferentiate to propagate the tumor bulk.

PubMed

Shukrun, Rachel; Pode-Shakked, Naomi; Pleniceanu, Oren; Omer, Dorit; Vax, Einav; Peer, Eyal; Pri-Chen, Sara; Jacob, Jasmine; Hu, Qianghua; Harari-Steinberg, Orit; Huff, Vicki; Dekel, Benjamin

2014-07-08

An open question remains in cancer stem cell (CSC) biology whether CSCs are by definition at the top of the differentiation hierarchy of the tumor. Wilms' tumor (WT), composed of blastema and differentiated renal elements resembling the nephrogenic zone of the developing kidney, is a valuable model for studying this question because early kidney differentiation is well characterized. WT neural cell adhesion molecule 1-positive (NCAM1(+)) aldehyde dehydrogenase 1-positive (ALDH1(+)) CSCs have been recently isolated and shown to harbor early renal progenitor traits. Herein, by generating pure blastema WT xenografts, composed solely of cells expressing the renal developmental markers SIX2 and NCAM1, we surprisingly show that sorted ALDH1(+) WT CSCs do not correspond to earliest renal stem cells. Rather, gene expression and proteomic comparative analyses disclose a cell type skewed more toward epithelial differentiation than the bulk of the blastema. Thus, WT CSCs are likely to dedifferentiate to propagate WT blastema.

Wilms’ Tumor Blastemal Stem Cells Dedifferentiate to Propagate the Tumor Bulk

PubMed Central

Shukrun, Rachel; Pode-Shakked, Naomi; Pleniceanu, Oren; Omer, Dorit; Vax, Einav; Peer, Eyal; Pri-Chen, Sara; Jacob, Jasmine; Hu, Qianghua; Harari-Steinberg, Orit; Huff, Vicki; Dekel, Benjamin

2014-01-01

Summary An open question remains in cancer stem cell (CSC) biology whether CSCs are by definition at the top of the differentiation hierarchy of the tumor. Wilms’ tumor (WT), composed of blastema and differentiated renal elements resembling the nephrogenic zone of the developing kidney, is a valuable model for studying this question because early kidney differentiation is well characterized. WT neural cell adhesion molecule 1-positive (NCAM1+) aldehyde dehydrogenase 1-positive (ALDH1+) CSCs have been recently isolated and shown to harbor early renal progenitor traits. Herein, by generating pure blastema WT xenografts, composed solely of cells expressing the renal developmental markers SIX2 and NCAM1, we surprisingly show that sorted ALDH1+ WT CSCs do not correspond to earliest renal stem cells. Rather, gene expression and proteomic comparative analyses disclose a cell type skewed more toward epithelial differentiation than the bulk of the blastema. Thus, WT CSCs are likely to dedifferentiate to propagate WT blastema. PMID:25068119

Influence of a Small Amount of Glycerol on the Trehalose Bioprotective Action Analyzed In Situ During Freeze-Drying of Lyzozyme Formulations by Micro-Raman Spectroscopy.

PubMed

Starciuc, Tatiana; Guinet, Yannick; Paccou, Laurent; Hedoux, Alain

2017-10-01

Micro-Raman spectroscopy gives the original opportunity to monitor simultaneously the operating process and the protein structure from in situ investigations along the 3 stages of the freeze-drying (FD) process. This opportunity was used for determining how a small amount of glycerol enhances the bioprotective efficiency of trehalose during FD of lysozyme formulations. Three lysozyme formulations were analyzed: lysozyme dissolved in D 2 O (wt% 1:9), in trehalose-D 2 O mixture (wt% 1:1:8), and in the trehalose-glycerol-D 2 O mixture (wt% 1:1:0.17:7.93). Raman mapping performed during each stage of the FD process has provided important information about the preferential interaction between water, trehalose, and lysozyme in relation to the protein stability. It was found that the addition of a small amount of glycerol had a plasticizing effect on the glassy trehalose-water matrix during the primary drying stage and then reduced the bioprotective effect of trehalose. By contrast, during the secondary drying stage, glycerol significantly enhanced the stabilizing effect of trehalose in the same sample, by replacing water-trehalose H-bonds with stronger glycerol-trehalose H-bonds and stiffening the amorphous trehalose matrix. The action of glycerol is also related to its capability to prevent aggregation of trehalose, making the structure of the frozen product more homogeneous, by changing the hydrogen-bond networks in the liquid formulation before the freezing stage. Copyright © 2017 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.

CHARACTERIZATION OF A PRECIPITATE REACTOR FEED TANK (PRFT) SAMPLE FROM THE DEFENSE WASTE PROCESSING FACILITY (DWPF)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Crawford, C.; Bannochie, C.

2014-05-12

A sample of from the Defense Waste Processing Facility (DWPF) Precipitate Reactor Feed Tank (PRFT) was pulled and sent to the Savannah River National Laboratory (SRNL) in June of 2013. The PRFT in DWPF receives Actinide Removal Process (ARP)/ Monosodium Titanate (MST) material from the 512-S Facility via the 511-S Facility. This 2.2 L sample was to be used in small-scale DWPF chemical process cell testing in the Shielded Cells Facility of SRNL. A 1L sub-sample portion was characterized to determine the physical properties such as weight percent solids, density, particle size distribution and crystalline phase identification. Further chemical analysismore » of the PRFT filtrate and dissolved slurry included metals and anions as well as carbon and base analysis. This technical report describes the characterization and analysis of the PRFT sample from DWPF. At SRNL, the 2.2 L PRFT sample was composited from eleven separate samples received from DWPF. The visible solids were observed to be relatively quick settling which allowed for the rinsing of the original shipping vials with PRFT supernate on the same day as compositing. Most analyses were performed in triplicate except for particle size distribution (PSD), X-ray diffraction (XRD), Scanning Electron Microscopy (SEM) and thermogravimetric analysis (TGA). PRFT slurry samples were dissolved using a mixed HNO3/HF acid for subsequent Inductively Coupled Plasma Atomic Emission Spectroscopy (ICPAES) and Inductively Coupled Plasma Mass Spectroscopy (ICP-MS) analyses performed by SRNL Analytical Development (AD). Per the task request for this work, analysis of the PRFT slurry and filtrate for metals, anions, carbon and base were primarily performed to support the planned chemical process cell testing and to provide additional component concentrations in addition to the limited data available from DWPF. Analysis of the insoluble solids portion of the PRFT slurry was aimed at detailed characterization of these solids (TGA, PSD, XRD and SEM) in support of the Salt IPT chemistry team. The overall conclusions from analyses performed in this study are that the PRFT slurry consists of 0.61 Wt.% insoluble MST solids suspended in a 0.77 M [Na+] caustic solution containing various anions such as nitrate, nitrite, sulfate, carbonate and oxalate. The corresponding measured sulfur level in the PRFT slurry, a critical element for determining how much of the PRFT slurry gets blended into the SRAT, is 0.437 Wt.% TS. The PRFT slurry does not contain insoluble oxalates nor significant quantities of high activity sludge solids. The lack of sludge solids has been alluded to by the Salt IPT chemistry team in citing that the mixing pump has been removed from Tank 49H, the feed tank to ARP-MCU, thus allowing the sludge solids to settle out. The PRFT aqueous slurry from DWPF was found to contain 5.96 Wt.% total dried solids. Of these total dried solids, relatively low levels of insoluble solids (0.61 Wt.%) were measured. The densities of both the filtrate and slurry were 1.05 g/mL. Particle size distribution of the PRFT solids in filtered caustic simulant and XRD analysis of washed/dried PRFT solids indicate that the PRFT slurry contains a bimodal distribution of particles in the range of 1 and 6 μm and that the particles contain sodium titanium oxide hydroxide Na2Ti2O4(OH)2 crystalline material as determined by XRD. These data are in excellent agreement with similar data obtained from laboratory sampling of vendor supplied MST. Scanning Electron Microscopy (SEM) combined with Energy Dispersive X-ray Spectroscopy (EDS) analysis of washed/dried PRFT solids shows the particles to be like previous MST analyses consisting of irregular shaped micron-sized solids consisting primarily of Na and Ti. Thermogravimetric analysis of the washed and unwashed PRFT solids shows that the washed solids are very similar to MST solids. The TGA mass loss signal for the unwashed solids shows similar features to TGA performed on cellulose nitrate filter paper indicating significant presence of the deteriorated filter in this unwashed sample. Neither the washed nor unwashed PRFT solids TGA traces showed any features that would indicate presence of sodium oxalate solids. The PRFT Filtrate elemental analysis shows that Na, S and Al are major soluble species with trace levels of B, Cr, Cu, K, Li, Si, Tc, Th and U present. Nitrate, nitrite, sulfate, oxalate, carbonate and hydroxide are major soluble anion species. There is good agreement between the analyzed TOC and the total carbon calculated from the sum of oxalate and minor species formate. Comparison of the amount and speciation of the carbon species between filtrate and slurry indicates no significant carbon-containing species, e.g., sodium oxalate, are present in the slurry solids. Dissolution of the PRFT slurry and subsequent analysis shows that Na, Ti, Si and U are the major elements present on a Wt.% total dried solids basis with 30, 5.8 and 0.47 and 0.11 Wt.% total dried solids, respectively. The amount of Al in the dissolved PRFT slurry is less than that calculated from the PRFT filtrate alone which suggests that the mixed acid digestion used in this work is not optimized for Al recovery. The concentrations of Ca, Fe, Hg and U are all low (at or below 0.11 wt%) and there is no detectable Mn or Ni present which indicates no significant HLW sludge solids are present in the PRFT slurry sample.« less

High titer ethanol production from SPORL-pretreated lodgepole pine by simultaneous enzymatic saccharification and combined fermentation

Treesearch

T.Q. Lan; Roland Gleisner; J.Y. Zhu; Bruce S. Dien; Ronald E. Hector

2012-01-01

Lodgepole wood chips were pretreated by sulfite pretreatment to overcome recalcitrance of lignocelluloses (SPORL) at 25% solids loading and 180 °C for 20 min with sulfuric acid and sodium bisulfite charges of 2.2 and 8 wt/wt% on an oven-dry wood basis, respectively. The pretreated wood chips were disk-milled with pretreatment spent liquor and water, and the...

Response of Pleurotus ostreatus to cadmium exposure

DOE Office of Scientific and Technical Information (OSTI.GOV)

Favero, N.; Bressa, G.; Costa, P.

1990-08-01

The possibility of utilizing agroindustrial wastes in the production of edible, high-quality products (e.g., mushrooms) implies the risk of bringing toxic substances, such as heavy metals, into the human food chain. Thus, growth in the presence of cadmium and cadmium accumulation limits have been studied in the industrially cultivated fungus P. ostreatus. Fruit body production is substantially unaffected in the presence of 25, 139, and 285 mg Cd/kg of dried substrate. Cadmium concentration in fruit bodies is related to cadmium substrate level, the metal being present at higher levels in caps (22-56 mg/kg dry wt) than in stems (13-36 mg/kgmore » dry wt). Concentration factor (CF), very low in the controls (about 2), further decreases in treated specimens. The presence of a cadmium control mechanism in this fungi species is suggested. Fruit body cadmium levels could, however, represent a risk for P. ostreatus consumers, according to FAO/WHO limits related to weekly cadmium intake.« less

The Transcription Factor Wilms Tumor 1 Confers Resistance in Myeloid Leukemia Cells against the Proapoptotic Therapeutic Agent TRAIL (Tumor Necrosis Factor α-related Apoptosis-inducing Ligand) by Regulating the Antiapoptotic Protein Bcl-xL*

PubMed Central

Bansal, Hima; Seifert, Theresea; Bachier, Carlos; Rao, Manjeet; Tomlinson, Gail; Iyer, Swaminathan Padmanabhan; Bansal, Sanjay

2012-01-01

Tumor necrosis factor α-related apoptosis-inducing ligand (TRAIL) is considered a promising cancer therapeutic agent due to its ability to induce apoptosis in a variety of cancer cells, while sparing normal cells. However, many human tumors including acute myeloid leukemia (AML) are partially or completely resistant to monotherapy with TRAIL, limiting its therapeutic utility. Therefore, identification of factors that contribute to TRAIL resistance may facilitate future development of more effective TRAIL-based cancer therapies. Here, we report a previously unknown role for WT1 in mediating TRAIL resistance in leukemia. Knockdown of WT1 with shRNA rendered TRAIL-resistant myeloid leukemia cells sensitive to TRAIL-induced cell death, and re-expression of shRNA-resistant WT1 restored TRAIL resistance. Notably, TRAIL-mediated apoptosis in WT1-silenced cells was largely due to down-regulation of the antiapoptotic protein Bcl-xL. Moreover, WT1 expression strongly correlated with overexpression of Bcl-xL in AML cell lines and blasts from AML patients. Furthermore, we found that WT1 transactivates Bcl-xL by directly binding to its promoter. We previously showed that WT1 is a novel client protein of heat shock protein 90 (Hsp90). Consistent with this, pharmacological inhibition of Hsp90 resulted in reduced WT1 and Bcl-xL expression leading to increased sensitivity of leukemia cells to TRAIL-mediated apoptosis. Collectively, our results suggest that WT1-dependent Bcl-xL overexpression contributes to TRAIL resistance in myeloid leukemias. PMID:22898820

The transcription factor Wilms tumor 1 confers resistance in myeloid leukemia cells against the proapoptotic therapeutic agent TRAIL (tumor necrosis factor α-related apoptosis-inducing ligand) by regulating the antiapoptotic protein Bcl-xL.

PubMed

Bansal, Hima; Seifert, Theresea; Bachier, Carlos; Rao, Manjeet; Tomlinson, Gail; Iyer, Swaminathan Padmanabhan; Bansal, Sanjay

2012-09-21

Tumor necrosis factor α-related apoptosis-inducing ligand (TRAIL) is considered a promising cancer therapeutic agent due to its ability to induce apoptosis in a variety of cancer cells, while sparing normal cells. However, many human tumors including acute myeloid leukemia (AML) are partially or completely resistant to monotherapy with TRAIL, limiting its therapeutic utility. Therefore, identification of factors that contribute to TRAIL resistance may facilitate future development of more effective TRAIL-based cancer therapies. Here, we report a previously unknown role for WT1 in mediating TRAIL resistance in leukemia. Knockdown of WT1 with shRNA rendered TRAIL-resistant myeloid leukemia cells sensitive to TRAIL-induced cell death, and re-expression of shRNA-resistant WT1 restored TRAIL resistance. Notably, TRAIL-mediated apoptosis in WT1-silenced cells was largely due to down-regulation of the antiapoptotic protein Bcl-xL. Moreover, WT1 expression strongly correlated with overexpression of Bcl-xL in AML cell lines and blasts from AML patients. Furthermore, we found that WT1 transactivates Bcl-xL by directly binding to its promoter. We previously showed that WT1 is a novel client protein of heat shock protein 90 (Hsp90). Consistent with this, pharmacological inhibition of Hsp90 resulted in reduced WT1 and Bcl-xL expression leading to increased sensitivity of leukemia cells to TRAIL-mediated apoptosis. Collectively, our results suggest that WT1-dependent Bcl-xL overexpression contributes to TRAIL resistance in myeloid leukemias.

Wt1 dictates the fate of fetal and adult Leydig cells during development in the mouse testis.

PubMed

Wen, Qing; Zheng, Qiao-Song; Li, Xi-Xia; Hu, Zhao-Yuan; Gao, Fei; Cheng, C Yan; Liu, Yi-Xun

2014-12-15

Wilms' tumor 1 (Wt1) is a tumor suppressor gene encoding ∼24 zinc finger transcription factors. In the mammalian testis, Wt1 is expressed mostly by Sertoli cells (SCs) involved in testis development, spermatogenesis, and adult Leydig cell (ALC) steroidogenesis. Global knockout (KO) of Wt1 is lethal in mice due to defects in embryogenesis. Herein, we showed that Wt1 is involved in regulating fetal Leydig cell (FLC) degeneration and ALC differentiation during testicular development. Using Wt1(-/flox);Amh-Cre mice that specifically deleted Wt1 in the SC vs. age-matched wild-type (WT) controls, FLC-like-clusters were found in Wt1-deficient testes that remained mitotically active from postnatal day 1 (P1) to P56, and no ALC was detected at these ages. Leydig cells in mutant adult testes displayed morphological features of FLC. Also, FLC-like cells in adult mutant testes had reduced expression in ALC-associated genes Ptgds, Sult1e1, Vcam1, Hsd11b1, Hsd3b6, and Hsd17b3 but high expression of FLC-associated genes Thbs2 and Hsd3b1. Whereas serum LH and testosterone level in mutant mice were not different from controls, intratesticular testosterone level was significantly reduced. Deletion of Wt1 gene also perturbed the expression of steroidogenic enzymes Star, P450c17, Hsd3b6, Hsd3b1, Hsd17b1, and Hsd17b3. FLCs in adult mutant testes failed to convert androstenedione to testosterone due to a lack of Hsd17b3, and this defect was rescued by coculturing with fetal SCs. In summary, FLC-like cells in mutant testes are putative FLCs that remain mitotically active in adult mice, illustrating that Wt1 dictates the fate of FLC and ALC during postnatal testis development. Copyright © 2014 the American Physiological Society.

Proteomic and Metabolic Analyses of S49 Lymphoma Cells Reveal Novel Regulation of Mitochondria by cAMP and Protein Kinase A*

PubMed Central

Wilderman, Andrea; Guo, Yurong; Divakaruni, Ajit S.; Perkins, Guy; Zhang, Lingzhi; Murphy, Anne N.; Taylor, Susan S.; Insel, Paul A.

2015-01-01

Cyclic AMP (cAMP), acting via protein kinase A (PKA), regulates many cellular responses, but the role of mitochondria in such responses is poorly understood. To define such roles, we used quantitative proteomic analysis of mitochondria-enriched fractions and performed functional and morphologic studies of wild-type (WT) and kin− (PKA-null) murine S49 lymphoma cells. Basally, 75 proteins significantly differed in abundance between WT and kin− S49 cells. WT, but not kin−, S49 cells incubated with the cAMP analog 8-(4-chlorophenylthio)adenosine cAMP (CPT-cAMP) for 16 h have (a) increased expression of mitochondria-related genes and proteins, including ones in pathways of branched-chain amino acid and fatty acid metabolism and (b) increased maximal capacity of respiration on branched-chain keto acids and fatty acids. CPT-cAMP also regulates the cellular rate of ATP-utilization, as the rates of both ATP-linked respiration and proton efflux are decreased in WT but not kin− cells. CPT-cAMP protected WT S49 cells from glucose or glutamine deprivation, In contrast, CPT-cAMP did not protect kin− cells or WT cells treated with the PKA inhibitor H89 from glutamine deprivation. Under basal conditions, the mitochondrial structure of WT and kin− S49 cells is similar. Treatment with CPT-cAMP produced apoptotic changes (i.e. decreased mitochondrial density and size and loss of cristae) in WT, but not kin− cells. Together, these findings show that cAMP acts via PKA to regulate multiple aspects of mitochondrial function and structure. Mitochondrial perturbation thus likely contributes to cAMP/PKA-mediated cellular responses. PMID:26203188

Effects of acid mine drainage from an abandoned copper mine, Britannia Mines, Howe Sound, British Columbia, Canada, on transplanted blue mussels (Mytilus edulis).

PubMed

Grout, J A; Levings, C D

2001-04-01

Juvenile mussels (Mytilus edulis) were transplanted to Howe Sound, British Columbia, Canada, along an apparent pollution gradient of acid mine drainage (AMD) from an abandoned copper (Cu) mine. Cages containing 75 mussels each were placed at a total of 15 stations and were exposed to concentrations of dissolved Cu in surface waters ranging from 5 to 1009 micrograms/l for a period of 41 days. Mussels located at stations closer to the source of AMD at the mouth of Britannia Creek bioaccumulated higher concentrations of Cu and zinc (Zn) in their tissues. Mussel growth was adversely affected by Cu tissue concentrations above 20 micrograms/g dry wt., while declines in survival and condition index occurred in mussels that bioaccumulated greater than 40 micrograms/g dry wt. Cu. Tissue Zn concentrations (117-192 micrograms/g dry wt.) were likely not high enough to have a direct impact on mussel health. Reduced survival of transplanted mussels was supported by an absence of natural mussels in contaminated areas. Phytoplankton was also severely reduced in areas contaminated by mine waters. Based on the weight of evidence, AMD from the Britannia mine had a deleterious impact on mussel survival in a zone extending at least 2.1 km to the north and 1.7 km to the south of Britannia Creek on the east shore of Howe Sound.

Seed development, seed germination and seedling growth in the R50 (sym16) pea mutant are not directly linked to altered cytokinin homeostasis.

PubMed

Long, Chengli; Held, Mark; Hayward, Allison; Nisler, Jaroslav; Spíchal, Lukas; Neil Emery, R J; Moffatt, Barbara A; Guinel, Frédérique C

2012-06-01

R50 (sym16) is a pea nodulation mutant that accumulates cytokinin (CK) in its vegetative organs. Total CK content increases as the plant ages because of the low activity of the enzyme cytokinin oxidase/dehydrogenase (CKX) responsible for CK degradation. R50 exhibits a large seed with high relative water content, and its seedling establishes itself slowly. Whether these two traits are linked to abnormal CK levels was considered here. R50 was found to have a similar germination rate but a much slower epicotyl emergence than Sparkle, its wild-type (WT). At the onset of emergence, the starch grains in R50 cotyledons were larger than those of WT; furthermore, they did not degrade as fast as in WT because of low amylase activity. No differences between the pea lines were observed in the CK forms identified during seed embryogenesis. However, while CK content compared to that of WT was reduced early in R50 embryogenesis, it was elevated later on in its dry seeds where CKX activity was low, although CKX transcript abundance remained high. Transcripts of the two known PsCKX isoforms exhibited tissue- and development-specific profiles with no detectable PsCKX2 expression in cotyledons. There were more of both transcripts in R50 roots than in WT roots, but less of PsCKX2 than PsCKX1 in R50 shoots compared to WT shoots. Thus, although there is a definite CKX post-transcriptional defect in R50 dry seeds, an abnormal CK homeostasis is not the basis of the delay in R50 seedling establishment, which we linked to abnormal amylase activity early in development. Copyright © Physiologia Plantarum 2012.

Preparation of laminated poly(ε-caprolactone)-gelatin-hydroxyapatite nanocomposite scaffold bioengineered via compound techniques for bone substitution

PubMed Central

Hamlekhan, Azhang; Moztarzadeh, Fathollah; Mozafari, Masoud; Azami, Mahmoud; Nezafati, Nader

2011-01-01

In this research, new bioactive nanocomposite scaffolds were successfully developed using poly(ε-caprolactone) (PCL), cross-linked gelatin and nanoparticles of hydroxyapatite (HAp) after testing different solvents and methods. First, HAp powder was synthesized via a chemical precipitation technique and characterized. Then, the nanocomposites were prepared through layer solvent casting combined with freeze-drying and lamination techniques. According to the results, the increasing of the PCL weight in the scaffolds led to the improvement of the mechanical properties. The amount of ultimate stress, stiffness and also elastic modulus increased from 8 MPa for 0% wt PCL to 23.5 MPa for 50% wt PCL. The biomineralization study revealed the formation of an apatite layer on the scaffolds after immersion in simulated body fluid (SBF). The Ca-P ratios were in accordance to nonstoichiometric biological apatite, which was approximately 1.67. The in vitro biocompatibility and cytocompatibility of the scaffolds were tested using mesenchymal stem cells (MSCs), and the results indicated no sign of toxicity, and cells were found to be attached to the scaffold walls. The in vivo biocompatibility and osteogenesis of these scaffolds in the animal experiments is also under investigation, and the result will be published at the end of the study. PMID:23507731

Identification of Methanotrophic Biomarker Lipids in the Symbiont-Containing Gills of Seep Mussels

NASA Technical Reports Server (NTRS)

Jahnke, L. L.; Zahiralis, K. D.; Klein, H. P.; Morrison, David (Technical Monitor)

1994-01-01

Mussels collected from hydrocarbon seeps in the Gulf of Mexico grow with methane as sole carbon and energy source due to a symbiotic association with methane-oxidizing bacteria. Transmission electron micrographs of mussel gills show cells with stacked intracytoplasmic membranes similar to type I methanotrophic bacteria. Methanotrophs are known to synthesize several types of cyclic triterpenes, hopanoids and methyl sterols, as well as unique monounsaturated fatty acid, double bond positional isomers that serve as biomarkers for this group. Lipid analysis of dissected mussels demonstrated the presence of these biomarkers predominantly in the gill tissue with much smaller amounts in mantle and remaining body tissues. Gill tissue contained 1150 micrograms/g dry wt. of hopanepolyol derivatives and diplopterol while the mantle tissue contained only 17 micrograms/g. The C16 monounsaturated fatty acids (16:1) characteristic of type I methanotrophic membranes dominated the gill tissue making up 53% of the total while only 5% 16:1 was present in the mantle tissue. The methyl sterol distribution was more dispersed. The predominant sterol in all tissues was cholesterol with lesser amounts of other desmethyl and 4-methyl sterols. The gill and mantle tissues contained 3461 micrograms (17% methyl) and 2750 micrograms (5% methyl) sterol per gm dry wt., respectively. Methyl sterol accounted for 44% of the sterol esters isolated from the gill, suggesting active demethylation of the methanotrophic sterols in this tissue. The use of lipid biomarkers could provide an effective means for identifying host-symbiont relationships.

Trace elements, organochlorines, polycyclic aromatic hydrocarbons, dioxins, and furans in lesser scaup wintering on the Indiana Harbor Canal

USGS Publications Warehouse

Custer, T.W.; Custer, Christine M.; Hines, R.K.; Sparks, D.W.

2000-01-01

During the winter of 1993a??94, male lesser scaup (Aythya affinis) were collected on the heavily polluted Indiana Harbor Canal (IHC), East Chicago, IN, USA, and examined for tissue contaminant levels. Lesser scaup collected on the IHC had higher concentrations of cadmium (Cd), mercury (Hg), selenium (Se), polychlorinated biphenyls (PCBs), selected organchlorine pesticides, polychlorinated dibenzodioxins, polychlorinated dibenzofurans, polycyclic aromatic hydrocarbons, and aliphatic hydrocarbons than reference birds. Of the scaup collected on the IHC, 44% had Cd concentrations in the liver considered above background for freshwater waterfowl (>3 I?g/g dry wt.), 50% had Se concentrations in the liver above a level possibly harmful to the health of young and adult birds (>33 I?g/g dry wt.), and 88% of the scaup carcasses exceeded the PCB human consumption guidelines for edible poultry in the USA (>3.0 I?g/g lipid wt.). The ratio of pristane:n-heptadecane concentrations in 47% of lesser scaup collected on IHC was elevated above 1.0, which is indicative of chronic exposure to petroleum hydrocarbons. Copyright A? 2009 Elsevier B.V.

Trace elements, organochlorines, polycyclic aromatic hydrocarbons, dioxins, and furans in lesser scaup wintering on the Indiana Harbor Canal.

PubMed

Custer, T W; Custer, C M; Hines, R K; Sparks, D W

2000-12-01

During the winter of 1993-94, male lesser scaup (Aythya affinis) were collected on the heavily polluted Indiana Harbor Canal (IHC), East Chicago, IN, USA, and examined for tissue contaminant levels. Lesser scaup collected on the IHC had higher concentrations of cadmium (Cd), mercury (Hg), selenium (Se), polychlorinated biphenyls (PCBs), selected organchlorine pesticides, polychlorinated dibenzodioxins, polychlorinated dibenzofurans, polycyclic aromatic hydrocarbons, and aliphatic hydrocarbons than reference birds. Of the scaup collected on the IHC, 44% had Cd concentrations in the liver considered above background for freshwater waterfowl (>3 microg/g dry wt.), 50% had Se concentrations in the liver above a level possibly harmful to the health of young and adult birds (>33 microg/g dry wt.), and 88% of the scaup carcasses exceeded the PCB human consumption guidelines for edible poultry in the USA (>3.0 microg/g lipid wt.). The ratio of pristane:n-heptadecane concentrations in 47% of lesser scaup collected on IHC was elevated above 1.0, which is indicative of chronic exposure to petroleum hydrocarbons.

Biopolymer-prebiotic carbohydrate blends and their effects on the retention of bioactive compounds and maintenance of antioxidant activity.

PubMed

Silva, Eric Keven; Zabot, Giovani L; Cazarin, Cinthia B B; Maróstica, Mário R; Meireles, M Angela A

2016-06-25

The objective of this study was to evaluate the use of inulin (IN), a prebiotic carbohydrate without superficial activity, as an encapsulating matrix of lipophilic bioactive compounds. For achieving the encapsulation, IN was associated with biopolymers that present superficial activity: modified starch (HiCap), whey protein isolate (WPI) and gum acacia (GA). Encapsulation was performed through emulsification assisted by ultrasound followed by freeze-drying (FD) process to dry the emulsions. All blends retained geranylgeraniol. GA-IN blend yielded the highest geranylgeraniol retention (96±2wt.%) and entrapment efficiency (94±3wt.%), whilst WPI-IN blend yielded the highest encapsulation efficiency (88±2wt.%). After encapsulation, composition of geranylgeraniol in the annatto seed oil was maintained (23.0±0.5g/100g of oil). Such findings indicate that the method of encapsulation preserved the active compound. All blends were also effective for maintaining the antioxidant activity of the oil through ORAC and DPPH analyses. Copyright © 2016 Elsevier Ltd. All rights reserved.

A Summary of Selected Data: DSDP Legs 1-19,

DTIC Science & Technology

1980-09-01

100 minerals may be applied in the future (densi ty water when the mineralogy and attenuation wt. water + wt. dry sed. + salt coefficients become...may be applied in the future when densities of some common minerals are the exact quantitative mineralogy and listed in Harms and Choquette (1965...calculation. These measurements different attenuation coefficient than were used to get a " ball park" answer that of calcite. for a particular sediment type

NOX2 Deficiency Protects Against Streptozotocin-Induced β-Cell Destruction and Development of Diabetes in Mice

PubMed Central

Xiang, Fu-Li; Lu, Xiangru; Strutt, Brenda; Hill, David J.; Feng, Qingping

2010-01-01

OBJECTIVE The role of NOX2-containing NADPH oxidase in the development of diabetes is not fully understood. We hypothesized that NOX2 deficiency decreases reactive oxygen species (ROS) production and immune response and protects against streptozotocin (STZ)-induced β-cell destruction and development of diabetes in mice. RESEARCH DESIGN AND METHODS Five groups of mice—wild-type (WT), NOX2−/−, WT treated with apocynin, and WT adoptively transferred with NOX2−/− or WT splenocytes—were treated with multiple-low-dose STZ. Blood glucose and insulin levels were monitored, and an intraperitoneal glucose tolerance test was performed. Isolated WT and NOX2−/− pancreatic islets were treated with cytokines for 48 h. RESULTS Significantly lower blood glucose levels, higher insulin levels, and better glucose tolerance was observed in NOX2−/− mice and in WT mice adoptively transferred with NOX2−/− splenocytes compared with the respective control groups after STZ treatment. Compared with WT, β-cell apoptosis, as determined by TUNEL staining, and insulitis were significantly decreased, whereas β-cell mass was significantly increased in NOX2−/− mice. In response to cytokine stimulation, ROS production was significantly decreased, and insulin secretion was preserved in NOX2−/− compared with WT islets. Furthermore, proinflammatory cytokine release induced by concanavalin A was significantly decreased in NOX2−/− compared with WT splenocytes. CONCLUSIONS NOX2 deficiency decreases β-cell destruction and preserves islet function in STZ-induced diabetes by reducing ROS production, immune response, and β-cell apoptosis. PMID:20627937

Optimized T-cell receptor-mimic chimeric antigen receptor T cells directed toward the intracellular Wilms Tumor 1 antigen

PubMed Central

Rafiq, S; Purdon, TJ; Daniyan, AF; Koneru, M; Dao, T; Liu, C; Scheinberg, DA; Brentjens, RJ

2017-01-01

CD19-directed chimeric antigen receptor (CAR) T cells are clinically effective in a limited set of leukemia patients. However, CAR T-cell therapy thus far has been largely restricted to targeting extracellular tumor-associated antigens (TAA). Herein, we report a T-cell receptor-mimic (TCRm) CAR, termed WT1-28z, that is reactive to a peptide portion of the intracellular onco-protein Wilms Tumor 1(WT1), as it is expressed on the surface of the tumor cell in the context of HLA-A*02:01. T cells modified to express WT1-28z specifically targeted and lysed HLA-A*02:01+ WT1+ tumors and enhanced survival of mice engrafted with HLA-A*02:01+, WT1+ leukemia or ovarian tumors. This in vivo functional validation of TCRm CAR T cells provides the proof-of-concept necessary to expand the range of TAA that can be effectively targeted for immunotherapy to include attractive intracellular targets, and may hold great potential to expand on the success of CAR T-cell therapy. PMID:27924074

Use of the yeast-like cells of Tremella fuciformis as a cell factory to produce a Pleurotus ostreatus hydrophobin.

PubMed

Zhu, Hanyu; Liu, Dongmei; Wang, Yuanyuan; Ren, Danfeng; Zheng, Liesheng; Chen, Liguo; Ma, Aimin

2017-08-01

To obtain hydrophobin, a Class I hydrophobin gene, Po.hyd from Pleurotus ostreatus, was transformed into the yeast-like cells of Tremella fuciformis using Agrobacterium tumefaciens. The hydrophobin Po.HYD from P. ostreatus was heterogeneously expressed by the yeast-like cells of T. fuciformis. Plasmids harboring the Po.hyd gene driven by endogenous glyceraldehyde-3-phosphate dehydrogenase promoter were transformed by A. tumefaciens. The integration and expression of the rPo.HYD in the T. fuciformis cells were confirmed by PCR, Southern blot, fluorescence microscopy and quantitative real-time PCR. SDS-PAGE demonstrated that the rPo.HYD was extracted with the expected MW of 14 kDa. The yield of purified rPo.HYD was 0.58 mg/g dry wt. The protein, with its ability to stabilize oil droplets, exhibited a better emulsifying activity than the typical food emulsifiers Tween 20 and sodium caseinate. Tremella fuciformis can be used as a cell factory to produce hydrophobin on a large scale for the food industry.

Immunohistochemical expression of WT1 by desmoplastic small round cell tumor: a comparative study with other small round cell tumors.

PubMed

Barnoud, R; Sabourin, J C; Pasquier, D; Ranchère, D; Bailly, C; Terrier-Lacombe, M J; Pasquier, B

2000-06-01

Desmoplastic small round cell tumors (DSRCTs) present a reciprocal chromosomal translocation, t(11;22)(p13;q12), that results in fusion of Ewing's sarcoma and Wilms' tumor (WT1) genes. The authors evaluated 15 DSRCTs and 71 other tumors often considered in the differential diagnosis for immunoreactivity using a polyclonal antibody directed against the WT1 part of the chimeric protein resulting from this translocation. WT1 immunostaining was performed on paraffin material using the WT(C-19) antibody after heat-antigen retrieval. All the DSRCTs (15 of 15, 100%) demonstrated strong WT1 nuclear immunoreactivity. Ten of 14 nephroblastomas (71%) disclosed WT1-positive nuclei in accordance with the staining reported by others, and rare and focal nuclear positivity was detected in two of 17 rhabdomyosarcomas. WT1 immunoreactivity was not observed in Ewing's sarcoma/primitive neuroectodermal tumors (zero of 21, 0%), neuroblastomas (zero of 17, 0%), or rhabdoid tumors of the kidney (zero of two, 0%). In nephroblastoma, differential diagnosis with DSRCT was not difficult: Clinical and morphologic data are not similar for these two entities. The current study validates WT1 immunoreactivity as a useful marker to separate DSRCT from other small round cell tumors.

[Characteristics of hydroxyapatite/PMMA nanocomposites for provisional restoration and its biocompatibility with human gingival fibroblasts].

PubMed

Zhang, Jing-chao; Mo, An-chun; Li, Ji-dong; Wang, Xue-jiang; Li, Yu-bao

2014-05-01

To formulate hydroxyapatite (HA)/polymethyl methacrylate (PMMA) composites with improved cytocompatibility for provisional restoration. Nanocomposites with 20 wt%, 30 wt%, 40 wt%, and 50 wt% HA/PMMA (H/P) were developed and examined using X-ray photoelectron spectroscopy (XPS) and scanning electron microscope (SEM). Human gingival fibroblasts were cultured on those HA/PMMA discs and investigated by fluorescent staining on 24 h and MTT assay at 1 d, 3 d, 5 d and 7 d. Chemical integration of HA/PMMA interface was confirmed by XPS. Typical fusiform cells with adhesion spots were detected on 40 wt% and 50 wt% H/P discs. MTT results showed insignificant differences in cell growth between 40 wt% H/P and pure titanium (Ti, P > 0.05), while the other H/P discs showed significantly lower cell growth than pure Ti (P < 0.05). 40 wt% H/P might be a promising candidate for provisional dental implant restoration and for esthetic gingival contour.

Filamin A regulates the organization and remodeling of the pericellular collagen matrix.

PubMed

Mezawa, Masaru; Pinto, Vanessa I; Kazembe, Mwayi P; Lee, Wilson S; McCulloch, Christopher A

2016-10-01

Extracellular matrix remodeling by cell adhesion-related processes is critical for proliferation and tissue homeostasis, but how adhesions and the cytoskeleton interact to organize the pericellular matrix (PCM) is not understood. We examined the role of the actin-binding protein, filamin A (FLNa), in pericellular collagen remodeling. Compared with wild-type (WT), mice with fibroblast-specific deletion of FLNa exhibited higher density but reduced organization of collagen fibers after increased loading of the periodontal ligament for 2 wk. In cultured fibroblasts, FLNa knockdown (KD) did not affect collagen mRNA, but after 24 h of culture, FLNa WT cells exhibited ∼2-fold higher cell-surface collagen KD cells and 13-fold higher levels of activated β1 integrins. In FLNa WT cells, there was 3-fold more colocalization of talin with pericellular cleaved collagen than in FLNa KD cells. MMP-9 mRNA and protein expression were >2-fold higher in FLNa KD cells than in WT cells. Cathepsin B, which is necessary for intracellular collagen digestion, was >3-fold higher in FLNa WT cells than in KD cells. FLNa WT cells exhibited 2-fold more collagen phagocytosis than KD cells, which involved the FLNa actin-binding domain. Evidently, FLNa regulates PCM remodeling through its effects on degradation pathways that affect the abundance and organization of collagen.-Mezawa, M., Pinto, V. I., Kazembe, M. P., Lee, W. S., McCulloch, C. A. Filamin A regulates the organization and remodeling of the pericellular collagen matrix. © FASEB.

Exercise metabolism in human skeletal muscle exposed to prior eccentric exercise

PubMed Central

Asp, Sven; Daugaard, Jens R; Kristiansen, Søren; Kiens, Bente; Richter, Erik A

1998-01-01

The effects of unaccustomed eccentric exercise on exercise metabolism during a subsequent bout of graded concentric exercise were investigated in seven healthy male subjects. Arterial and bilateral femoral venous catheters were inserted 2 days after eccentric exercise of one thigh (eccentric thigh) and blood samples were taken before and during graded two-legged concentric knee-extensor exercise. Muscle biopsies were obtained from the eccentric and control vastus lateralis before (rest) and after (post) the concentric exercise bout. Maximal knee-extensor concentric exercise capacity was decreased by an average of 23 % (P < 0.05) in the eccentric compared with the control thigh. The resting muscle glycogen content was lower in the eccentric thigh than in the control thigh (402 ± 30 mmol (kg dry wt)−1vs. 515 ± 26 mmol (kg dry wt)−1, means ± s.e.m., P < 0.05), and following the two-legged concentric exercise this difference substantially increased (190 ± 46 mmol (kg dry wt)−1vs. 379 ± 58 mmol (kg dry wt)−1, P < 0.05) despite identical power and duration of exercise with the two thighs. There was no measurable difference in glucose uptake between the eccentric and control thigh before or during the graded two-legged concentric exercise. Lactate release was higher from the eccentric thigh at rest and, just before termination of the exercise bout, release of lactate decreased from this thigh (suggesting decreased glycogenolysis), whereas no decrease was found from the contralateral control thigh. Lower glycerol release from the eccentric thigh during the first, lighter part of the exercise (P < 0.05) suggested impaired triacylglycerol breakdown. At rest, sarcolemmal GLUT4 glucose transporter content and glucose transport were similar in the two thighs, and concentric exercise increased sarcolemmal GLUT4 content and glucose transport capacity similarly in the two thighs. It is concluded that in muscle exposed to prior eccentric contractions, exercise at a given power output requires a higher relative workload than in undamaged muscle. This increases utilization of the decreased muscle glycogen stores, contributing to decreased endurance. PMID:9547403

Ecotoxicological study of arsenic and lead contaminated soils in former orchards at the Hanford Site, USA.

PubMed

Delistraty, Damon; Yokel, Jerry

2014-01-01

The purpose of this study was to assess ecotoxicity of former orchard soils contaminated with lead arsenate pesticides at the Hanford Site in Washington state (USA). Surface soil, plant, and invertebrate samples were collected from 11 sites in former orchard areas. Mean (standard deviation [SD]) for As and Pb in soil were 39.5 (40.6) and 208 (142) mg/kg dry wt, respectively (n = 11). These concentrations exceeded Hanford background levels but were similar to orchard soils elsewhere. In our study, As and Pb soil concentrations were positively and significantly correlated (r = 0.87, Bonferroni P < 0.05). Speciation of total inorganic As in soil (n = 6) demonstrated that As+5 was the dominant form (>99%). Mean (SD) for As and Pb in cheatgrass were 3.9 (7.9) and 12.4 (20.0) mg/kg dry wt, respectively (n = 11), while mean (SD) for As and Pb in darkling beetles were 5.4 (2.6) and 3.9 (3.0) mg/kg dry wt, respectively (n = 8). Linear regressions were constructed to estimate soil to cheatgrass and soil to darkling beetle uptake for As and Pb. These were significant (Bonferroni P < 0.05) only for cheatgrass versus soil (As) and darkling beetle versus soil (Pb). Standardized lettuce seedling and earthworm bioassays were performed with a subset of soil samples (n = 6). No significant effects (P > 0.05) were observed in lettuce survival or growth nor in earthworm survival or sublethal effects. Based on these bioassays, unbounded no observed effect concentrations (NOECs) in soil for As and Pb were 128 and 390 mg/kg dry wt, respectively. However, our range of soil concentrations generally overlapped a set of ecotoxicological benchmarks reported in the literature. Given uncertainty and limited sampling related to our NOECs, as well as uncertainty in generic benchmarks from the literature, further study is needed to refine characterization of As and Pb ecotoxicity in former orchard soils at the Hanford Site. Copyright © 2011 Wiley Periodicals, Inc.

Chronic toxicity of copper to five benthic invertebrates in laboratory-formulated sediment: sensitivity comparison and preliminary risk assessment.

PubMed

Roman, Yblin E; De Schamphelaere, Karel A C; Nguyen, Lien T H; Janssen, Colin R

2007-11-15

Five benthic organisms commonly used for sediment toxicity testing were chronically (28 to 35 days) exposed to copper in standard laboratory-formulated sediment (following Organization for Economic Cooperation and Development guidelines) and lethal and sub-lethal toxicities were evaluated. Sub-lethal endpoints considered were reproduction and biomass production for Lumbriculus variegatus, growth and reproduction for Tubifex tubifex, growth and emergence for Chironomus riparius, and growth for Gammarus pulex and Hyalella azteca. Expressed on whole-sediment basis the observed lethal sensitivity ranking (from most to least sensitive) was: G. pulex>L. variegatus>H. azteca=C. riparius=T. tubifex, with median chronic lethal concentrations (LC50) between 151 and 327 mg/kg dry wt. The sub-lethal sensitivity ranking (from most to least sensitive, with the most sensitive endpoint between parentheses): C. riparius (emergence)>T. tubifex (reproduction)=L. variegatus (reproduction)>G. pulex (growth)>H. azteca (growth), with median effective concentrations (EC50) between 59.2 and 194 mg/kg dry wt. No observed effect concentrations (NOEC) or 10% effective concentrations (EC10) for the five benthic invertebrates were used to perform a preliminary risk assessment for copper in freshwater sediment by means of (a) the "assessment factor approach" or (b) the statistical extrapolation approach (species sensitivity distribution). Depending on the data (NOEC or EC10) and the methodology used, we calculated a Predicted No Effect Concentration (PNEC) for sediment between 3.3 and 47.1 mg Cu/dry wt. This range is similar to the range of natural (geochemical) background concentrations of copper in sediments in Europe, i.e. 90% of sediments have a concentration between 5 and 49 mg Cu/kg dry wt. A detailed analysis of the outcome of this preliminary exercise highlighted that multiple issues need to be explored for achieving a scientifically more sound risk assessment and for the development of robust sediment quality criteria for copper, including (i) the use of the assessment factor approach vs. the statistical extrapolation approach, (ii) the importance of bioavailability modifying factors (e.g., organic carbon, acid volatile sulfide), and (iii) the influence of prevailing geochemical (bioavailable) background concentrations on the copper sensitivity of local benthic biota.

Geospatial association of endemicity of ataxic polyneuropathy and highly cyanogenic cassava cultivars.

PubMed

Oluwole, Olusegun Steven Ayodele; Oludiran, Adeyinka

2013-09-14

Exposure to cyanide from cassava foods is present in communities where ataxic polyneuropathy is endemic. Ataxic polyneuropathy is endemic in coastal parts of southwest and southeast Nigeria, and coastal Newala, south India, but it has been reported in epidemic or endemic forms from Africa, Asia, or Caribbean. This study was done to determine if cyanogenicity of cassava cultivars is higher in lowland than highland areas, and if areas of endemicity of ataxic polyneuropathy colocalize with areas of highest cyanogenicity of cassava. Roots of cassava cultivars were collected from 150 farmers in 32 of 37 administrative areas in Nigeria. Global positioning system was used to determine the location of the roots. Roots were assayed for concentrations of cyanogens. Thin Plate Spline regression was used to produce the contour map of cyanogenicity of the study area. Contour maps of altitude of the endemic areas were produced. Relationship of cyanogenicity of cassava cultivars and altitude, and of locations of areas of high cyanogenicity and areas of endemicity were determined. Geometrical mean (95% CI) cyanogen concentration was 182 (142-233) mg HCN eq/kg dry wt for cassava cultivars in areas ≤ 25 m above sea level, but 54 (43-66) mg HCN eq/kg dry wt for areas > 375 m. Non-spatial linear regression of altitude on logarithm transformed concentrations of cyanogens showed highly significant association, (p < 0.0001). Contour map of concentrations of cyanogens in cassava cultivars in Nigeria showed four areas with average concentrations of cassava cyanogens > 250 mg HCN eq/kg dry wt, and one area of moderately high cyanogen concentration > 150 mg HCN eq/kg dry wt. The endemic areas colocalized with areas of highest cassava cyanogenicity in lowland areas close to the Atlantic Ocean. This study shows strong geospatial association of areas of endemicity of ataxic polyneuropathy and areas of highest cyanogenicity of cassava cultivars. Finding of higher cyanogenicity of cassava in lowland than highland areas indicate strong influence of altitude on expression of cyanogens in cassava cultivars.

Geospatial association of endemicity of ataxic polyneuropathy and highly cyanogenic cassava cultivars

PubMed Central

2013-01-01

Background Exposure to cyanide from cassava foods is present in communities where ataxic polyneuropathy is endemic. Ataxic polyneuropathy is endemic in coastal parts of southwest and southeast Nigeria, and coastal Newala, south India, but it has been reported in epidemic or endemic forms from Africa, Asia, or Caribbean. This study was done to determine if cyanogenicity of cassava cultivars is higher in lowland than highland areas, and if areas of endemicity of ataxic polyneuropathy colocalize with areas of highest cyanogenicity of cassava. Methods Roots of cassava cultivars were collected from 150 farmers in 32 of 37 administrative areas in Nigeria. Global positioning system was used to determine the location of the roots. Roots were assayed for concentrations of cyanogens. Thin Plate Spline regression was used to produce the contour map of cyanogenicity of the study area. Contour maps of altitude of the endemic areas were produced. Relationship of cyanogenicity of cassava cultivars and altitude, and of locations of areas of high cyanogenicity and areas of endemicity were determined. Results Geometrical mean (95% CI) cyanogen concentration was 182 (142–233) mg HCN eq/kg dry wt for cassava cultivars in areas ≤ 25 m above sea level, but 54 (43–66) mg HCN eq/kg dry wt for areas > 375 m. Non-spatial linear regression of altitude on logarithm transformed concentrations of cyanogens showed highly significant association, (p < 0.0001). Contour map of concentrations of cyanogens in cassava cultivars in Nigeria showed four areas with average concentrations of cassava cyanogens > 250 mg HCN eq/kg dry wt, and one area of moderately high cyanogen concentration > 150 mg HCN eq/kg dry wt. The endemic areas colocalized with areas of highest cassava cyanogenicity in lowland areas close to the Atlantic Ocean. Conclusion This study shows strong geospatial association of areas of endemicity of ataxic polyneuropathy and areas of highest cyanogenicity of cassava cultivars. Finding of higher cyanogenicity of cassava in lowland than highland areas indicate strong influence of altitude on expression of cyanogens in cassava cultivars. PMID:24034556

Biomass production and nutrient removal potential of water hyacinth cultured in sewage effluent

DOE Office of Scientific and Technical Information (OSTI.GOV)

Reddy, K.R.; Hueston, F.M.; McKim, T.

1985-05-01

Growth and nutrient uptake of water hyacinth (Eichhornia crassipes (Mart) Solms) cultured in sewage effluent were measured over a period of 1 year in a prototype wastewater treatment system which has been in operation at Walt Disney World near Orlando, Florida. Annual productivity of water hyacinth cultured in primary sewage effluent (Channel II) was found to be in the range of 5 to 27 g dry wt m/sup -2/ day/sup -1/ (23.6 dry tons acre/sup -1/ yr/sup -1/). Average growth rate during the months of May through October 1982 for hyacinth cultured in Channel II (primary sewage effluent) and Channelmore » I (treated primary sewage effluent leaving Channel II) was about 16 g dry wt m/sup -2/ day/sup -1/ (27 dry tons acre/sup -1/ yr/sup -1/), compared to the growth rate of 13 g dry wt m/sup -2/ day/sup -1/ (22 dry tons acre/sup -1/ yr/sup -1/) for hyacinths cultured in secondary sewage effluent. Plants cultured in secondary sewage effluent generally had longer roots than the plants cultured in primary sewage effluent. A significant relationship was observed between the growth rate of hyacinth and the solar radiation. Nitrogen and P concentration of the plant tissue were higher in the hyacinths cultured during winter months compared to the plants grown in summer months. Average N and P concentration of the plants cultured in primary sewage effluent were found to be 3.7 percent N and 0.94 percent P, respectively, while the plants cultured in secondary sewage effluent had a total N and P content of 2.8 percent N and 0.79 percent P. Nutrient ratios of the major plant nutrients were found to be approximately the same as the nutrient ratios in the sewage effluent. Annual N and P uptake rates of hyacinth cultured in sewage effluent were found to be in the range of 1176 to 1193 kg N ha/sup -1/ yr/sup -1/ and 321 to 387 kg P ha/sup -1/ yr/sup -1/, respectively.« less

PTP1B triggers integrin-mediated repression of myosin activity and modulates cell contractility

PubMed Central

González Wusener, Ana E.; González, Ángela; Nakamura, Fumihiko; Arregui, Carlos O.

2016-01-01

ABSTRACT Cell contractility and migration by integrins depends on precise regulation of protein tyrosine kinase and Rho-family GTPase activities in specific spatiotemporal patterns. Here we show that protein tyrosine phosphatase PTP1B cooperates with β3 integrin to activate the Src/FAK signalling pathway which represses RhoA-myosin-dependent contractility. Using PTP1B null (KO) cells and PTP1B reconstituted (WT) cells, we determined that some early steps following cell adhesion to fibronectin and vitronectin occurred robustly in WT cells, including aggregation of β3 integrins and adaptor proteins, and activation of Src/FAK-dependent signalling at small puncta in a lamellipodium. However, these events were significantly impaired in KO cells. We established that cytoskeletal strain and cell contractility was highly enhanced at the periphery of KO cells compared to WT cells. Inhibition of the Src/FAK signalling pathway or expression of constitutive active RhoA in WT cells induced a KO cell phenotype. Conversely, expression of constitutive active Src or myosin inhibition in KO cells restored the WT phenotype. We propose that this novel function of PTP1B stimulates permissive conditions for adhesion and lamellipodium assembly at the protruding edge during cell spreading and migration. PMID:26700725

Comparison of blueberry powder produced via foam-mat freeze-drying versus spray-drying: evaluation of foam and powder properties.

PubMed

Darniadi, Sandi; Ho, Peter; Murray, Brent S

2018-03-01

Blueberry juice powder was developed via foam-mat freeze-drying (FMFD) and spray-drying (SD) via addition of maltodextrin (MD) and whey protein isolate (WPI) at weight ratios of MD/WPI = 0.4 to 3.2 (with a fixed solids content of 5 wt% for FMFD and 10 wt% for SD). Feed rates of 180 and 360 mL h -1 were tested in SD. The objective was to evaluate the effect of the drying methods and carrier agents on the physical properties of the corresponding blueberry powders and reconstituted products. Ratios of MD/WPI = 0.4, 1.0 and 1.6 produced highly stable foams most suitable for FMFD. FMFD gave high yields and low bulk density powders with flake-like particles of large size that were also dark purple with high red values. SD gave low powder recoveries. The powders had higher bulk density and faster rehydration times, consisting of smooth, spherical and smaller particles than in FMFD powders. The SD powders were bright purple but less red than FMFD powders. Solubility was greater than 95% for both FMFD and SD powders. The FMFD method is a feasible method of producing blueberry juice powder and gives products retaining more characteristics of the original juice than SD. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

Characterization and optimization of hydrogen production by a salt water blue-green alga Oscillatoria sp. Miami BG 7. II - Use of immobilization for enhancement of hydrogen production

NASA Technical Reports Server (NTRS)

Phlips, E. J.; Mitsui, A.

1986-01-01

The technique of cellular immobilization was applied to the process of hydrogen photoproduction of nonheterocystous, filamentous marine blue-green alga, Oscillatoria sp. Miami BG 7. Immobilization with agar significantly improved the rate and longevity of hydrogen production, compared to free cell suspensions. Rates of H2 production in excess of 13 microliters H2 mg dry/wt h were observed and hydrogen production was sustained for three weeks. Immobilization also provided some stabilization to environmental variability and was adaptable to outdoor light conditions. In general, immobilization provides significant advantages for the production and maintenance of hydrogen photoproduction for this strain.

Tissue lead distribution and hematologic effects in American kestrels (Falco sparverius) fed biologically incorporated lead

USGS Publications Warehouse

Custer, T.W.; Franson, J.C.; Pattee, O.H.

1984-01-01

American kestrels were fed a diet containing 0.5, 120, 212, and 448 ppm (dry wt) biologically incorporated lead (Pb) for 60 days. The diet consisted of homogenized 4-wk-old cockerels raised on feed mixed with and without lead. No kestrels died and weights did not differ among treatment groups. The control group (0.5 ppm Pb) had the lowest mean concentration of lead and the high dietary group had the highest for the following tissues: Kidney, liver, femur, brain, and blood. Concentrations of lead were significantly correlated among tissues. There were no differences among treatment groups for packed cell volume, hemoglobin concentration, or erythrocyte count.

Gel electrolytes with I-/I3- redox mediator based on methylcellulose for dye-sensitized solar cells

NASA Astrophysics Data System (ADS)

Yusof, S. Z.; Woo, H. J.; Careem, M. A.; Arof, A. K.

2018-05-01

A new gel electrolyte comprising methylcellulose (MC), LiBOB and succinonitrile (SN) has been prepared with dimethyl sulfoxide (DMSO) as solvent. The electrolyte with composition 8.73 wt % MC-2.92 wt % LiBOB-1.01 wt % SN-87.34 wt % DMSO exhibits the highest conductivity of 1.18 mS cm-1 at 25 °C. On partially substituting LiBOB with TMAI, the sample designated as TMAI 95 has the highest conducting composition of 8.70 wt % MC-0.14 wt % LiBOB-1.01 wt % SN-2.77 wt % TMAI-0.35 wt % I2-87.03 wt % DMSO. The conductivity is 1.96 mS cm-1. This sample is used to fabricate a dye sensitized photovoltaic cell that converts photons to electricity at an efficiency of 3.46%. The conductivity of this sample has been enhanced to 3.08 mS cm-1 on addition of 1.0 wt % butyl-methyl immidazolium iodide (BMII) ionic liquid and the efficiency of the cell fabricated is 4.63%. Total replacement of LiBOB component in the electrolyte with the same amount of LiI results in a conductivity increase of ∼23.5% and the DSSC exhibits a 5.72% efficiency.

BCAP inhibits proliferation and differentiation of myeloid progenitors in the steady state and during demand situations.

PubMed

Duggan, Jeffrey M; Buechler, Matthew B; Olson, Rebecca M; Hohl, Tobias M; Hamerman, Jessica A

2017-03-16

B-cell adaptor for phosphatidylinositol 3-kinase (BCAP) is a signaling adaptor expressed in mature hematopoietic cells, including monocytes and neutrophils. Here we investigated the role of BCAP in the homeostasis and development of these myeloid lineages. BCAP -/- mice had more bone marrow (BM) monocytes than wild-type (WT) mice, and in mixed WT:BCAP -/- BM chimeras, monocytes and neutrophils skewed toward BCAP -/- origin, showing a competitive advantage for BCAP -/- myeloid cells. BCAP was expressed in BM hematopoietic progenitors, including lineage - Sca-1 + c-kit + (LSK), common myeloid progenitor, and granulocyte/macrophage progenitor (GMP) cells. At the steady state, BCAP -/- GMP cells expressed more IRF8 and less C/EBPα than did WT GMP cells, which correlated with an increase in monocyte progenitors and a decrease in granulocyte progenitors among GMP cells. Strikingly, BCAP -/- progenitors proliferated and produced more myeloid cells of both neutrophil and monocyte/macrophage lineages than did WT progenitors in myeloid colony-forming unit assays, supporting a cell-intrinsic role of BCAP in inhibiting myeloid proliferation and differentiation. Consistent with these findings, during cyclophosphamide-induced myeloablation or specific monocyte depletion, BCAP -/- mice replenished circulating monocytes and neutrophils earlier than WT mice. During myeloid replenishment after cyclophosphamide-induced myeloablation, BCAP -/- mice had increased LSK proliferation and increased numbers of LSK and GMP cells compared with WT mice. Furthermore, BCAP -/- mice accumulated more monocytes and neutrophils in the spleen than did WT mice during Listeria monocytogenes infection. Together, these data identify BCAP as a novel inhibitor of myelopoiesis in the steady state and of emergency myelopoiesis during demand conditions. © 2017 by The American Society of Hematology.

The effects of 3% diquafosol sodium application on the tear functions and ocular surface of the Cu,Zn-superoxide dismutase-1 (Sod1)-knockout mice.

PubMed

Kojima, Takashi; Dogru, Murat; Ibrahim, Osama M; Nagata, Taeko; Higa, Kazunari; Shimizu, Takahiko; Shirasawa, Takuji; Satake, Yoshiyuki; Shimazaki, Seika; Shimazaki, Jun; Tsubota, Kazuo

2014-01-01

To investigate the role of a water and mucin secretagogue (3% diquafosol sodium eye drops) on the tear function and conjunctival ocular surface changes in Sod1(-/-) in comparison to the wild-type (WT) mice. Fourteen eyes of 7 Sod1(-/-) male mice with C57BL/background and 14 eyes of 7 C57BL6 strain wild-type male mice were examined at 40 weeks in this study. All mice had application of 3% diquafosol ophthalmic solution six times a day for 2 weeks. Tear film stability and corneal epithelial damage was evaluated by fluorescein and Rose Bengal stainings. Anterior segment photography was performed before and after eye drop instillations. Aqueous tear quantity was measured with phenol red-impregnated cotton threads without anesthesia. Animals were sacrificed at 42 weeks after diquafosol treatment and the whole globe specimens were subjected to periodic acid Schiff staining. Goblet cell density was quantified by J Image software. Quantitative real-time PCR for conjunctival muc 5AC messenger RNA expression was also performed. Sod1(-/-) mice had significantly higher fluorescein staining scores compared to the WT mice before eye drop instillation. The mean tear film breakup time, Rose Bengal staining scores, and muc5 messenger RNA expression improved significantly with diquafosol treatment in both the WT and the knockout mice. The mean fluorescein staining score and aqueous tear quantity significantly improved in the Sod1(-/-) mice with treatment. A notable and consistent increase in goblet cells and decrease in inflammatory cell infiltrates could be confirmed in all specimens after 2 weeks of diquafosol eye drop application. Three percent diquafosol ophthalmic solution appears to be effective in the treatment of ocular surface disease in this age-related dry eye disease mouse model.

Study on the Mechanism of Cell Cycle Checkpoint Kinase 2 (CHEK2) Gene Dysfunction in Chemotherapeutic Drug Resistance of Triple Negative Breast Cancer Cells.

PubMed

Luo, Li; Gao, Wei; Wang, Jinghui; Wang, Dingxue; Peng, Xiaobo; Jia, Zhaoyang; Jiang, Ye; Li, Gongzhuo; Tang, Dongxin; Wang, Yajie

2018-05-15

BACKGROUND This study aimed to investigate the mechanism of CHEK2 gene dysfunction in drug resistance of triple negative breast cancer (TNBC) cells. MATERIAL AND METHODS To perform our study, a stable CHEK2 wild type (CHEK2 WT) or CHEK2 Y390C mutation (CHEK2 Y390C) expressed MDA-MB-231 cell line was established. MTT assay, cell apoptosis assay and cell cycle assay were carried out to analyze the cell viability, apoptosis, and cell cycle respectively. Western blotting and qRT-PCR were applied for related protein and gene expression detection. RESULTS We found that the IC50 value of DDP (Cisplatin) to CHEK2 Y390C expressed MDA-MB-231 cells was significantly higher than that of the CHEK2 WT expressed cells and the control cells. After treatment with DDP for 48 h, cells expressing CHEK2 WT showed lower cell viability than that of the CHEK2 Y390C expressed cells and the control cells; compared with the CHEK2 Y390C expressed cells and the control cells, cells expressing CHEK2 WT showed significant G1/S arrest. Meanwhile, we found that compared with the CHEK2 Y390C expressed cells and the control cells, cell apoptosis was significantly increased in CHEK2 WT expressed cells. Moreover, our results suggested that cells expressing CHEK2 WT showed higher level of p-CDC25A, p-p53, p21, Bax, PUMA, and Noxa than that of the CHEK2 Y390C expressed cells and the control cells. CONCLUSIONS Our findings indicated that CHEK2 Y390C mutation induced the drug resistance of TNBC cells to chemotherapeutic drugs through administrating cell apoptosis and cell cycle arrest via regulating p53 activation and CHEK2-p53 apoptosis pathway.

Dry Sliding Tribological Studies of AA6061-B4C-Gr Hybrid Composites

NASA Astrophysics Data System (ADS)

Monikandan, V. V.; Joseph, M. A.; Rajendrakumar, P. K.

2016-10-01

The dry sliding behavior of stir-cast AA6061-10 wt.% B4C composites containing 2.5, 5 and 7.5 wt.% graphite particles was studied as a function of applied load, sliding speed and sliding distance on a pin-on-disk tribotester. The wear rate and friction coefficient increased with increase in applied load and sliding distance. The increase in graphite addition reduced the increase in wear rate and friction coefficient in the sliding speed range 2-2.5 m/s. Scanning electron microscopy of the worn pin revealed a graphite tribolayer, and transmission electron microscopy revealed overlapping deformation bands under 30 N applied load. Upon increasing the applied load to 40 N, welded region with fine crystalline structure was formed due to dynamic recrystallization of AA6061 alloy matrix.

Incorporation of Multiwalled Carbon Nanotubes into High Temperature Resin Using Dry Mixing Techniques

NASA Technical Reports Server (NTRS)

Ghose, Sayata; Watson, Kent A.; Delozier, Donavon M.; Working, Dennis C.; Siochi, Emilie J.; Connell, John W.

2006-01-01

As part of an ongoing effort to develop multifunctional advanced composites, blends of PETI330 and multiwalled carbon nanotubes (MWNTs) were prepared and characterized. Dry mixing techniques were employed and the maximum loading level of the MWNT chosen was based primarily on its effect on melt viscosity. The PETI330/ MWNT mixtures were prepared at concentrations ranging from 3 to 25 wt %. The resulting powders were characterized for homogeneity, thermal and rheological properties and extrudability as continuous fibers. Based on the characterization results, samples containing 10, 15 and 20 wt % MWNTs were chosen for more comprehensive evaluation. Samples were also prepared using in situ polymerization and solution mixing techniques and their properties were compared with the ball-mill prepared samples. The preparation and characterization of PETI330/ MWNT nanocomposites are discussed herein.

The Novel α4B Murine α4 Integrin Protein Splicing Variant Inhibits α4 Protein-dependent Cell Adhesion*

PubMed Central

Kouro, Hitomi; Kon, Shigeyuki; Matsumoto, Naoki; Miyashita, Tomoe; Kakuchi, Ayaka; Ashitomi, Dai; Saitoh, Kodai; Nakatsuru, Takuya; Togi, Sumihito; Muromoto, Ryuta; Matsuda, Tadashi

2014-01-01

Integrins affect the motility of multiple cell types to control cell survival, growth, or differentiation, which are mediated by cell-cell and cell-extracellular matrix interactions. We reported previously that the α9 integrin splicing variant, SFα9, promotes WT α9 integrin-dependent adhesion. In this study, we introduced a new murine α4 integrin splicing variant, α4B, which has a novel short cytoplasmic tail. In inflamed tissues, the expression of α4B, as well as WT α4 integrin, was up-regulated. Cells expressing α4B specifically bound to VCAM-1 but not other α4 integrin ligands, such as fibronectin CS1 or osteopontin. The binding of cells expressing WT α4 integrin to α4 integrin ligands is inhibited by coexpression of α4B. Knockdown of α4B in metastatic melanoma cell lines results in a significant increase in lung metastasis. Expression levels of WT α4 integrin are unaltered by α4B, with α4B acting as a regulatory subunit for WT α4 integrin by a dominant-negative effect or inhibiting α4 integrin activation. PMID:24755217

Distribution and speciation of mercury in the peat bog of Xiaoxing'an Mountain, northeastern China.

PubMed

Liu, Ruhai; Wang, Qichao; Lu, Xianguo; Fang, Fengman; Wang, Yan

2003-01-01

Most reports on mercury (Hg) in boreal ecosystems are from the Nordic countries and North America. Comparatively little information is available on Hg in wetlands in China. We present here a study on Hg in the Tangwang River forested catchment of the Xiaoxing'an Mountain in the northeast of China. The average total Hg (THg) in peat profile ranged from 65.8 to 186.6 ng g(-1) dry wt with the highest at the depth of 5-10 cm. THg in the peat surface was higher than the background in Heilongjiang province, the Florida Everglades, and Birkeness in Sweden. MethylHg (MeHg) concentration ranged from 0.16 to 1.86 ng g(-1) dry wt, with the highest amount at 10-15 cm depth. MeHg content was 0.2-1.2% of THg. THg and MeHg all decreased with the depth. THg in upland layer of soil (0-20 cm) was comparable to the peat surface, but in deeper layers THg concentration in peat was much higher than that in the forested mineral soil. THg in the peat bog increased, but MeHg decreased after it was drained. THg content in plant was different; THg contents in moss (119 ng g(-1) dry wt, n=12) were much higher than in the herbage, the arbor, and the shrubs. The peat bog has mainly been contaminated by Hg deposition from the atmosphere.

Biochemical composition of three species of unionid mussels after emersion

USGS Publications Warehouse

Greseth, Shari L.; Cope, W.G.; Rada, R.G.; Waller, D.L.; Bartsch, M.R.

2003-01-01

Freshwater mussels are emersed (exposed to air) during conservation activities such as surveys and relocations. Success of these activities depends upon the ability of mussels to survive emersion and to re-burrow in the substratum. We evaluated the acute sublethal effects of emersion on three species of unionid mussels [pocketbook, Lampsilis cardium (Rafinesque, 1820); pimpleback, Quadrula pustulosa pustulosa (I. Lea, 1831); spike, Elliptio dilatata (Rafinesque, 1820)] by measuring three biochemicals (carbohydrate, lipid, protein) indicative of biochemical function and energy storage. Mussels were acclimated in water at 25??C and exposed to five air temperatures (15, 20, 25, 35 and 45??C) for 15, 30 and 60 min. After emersion, mussels were returned to water at 25??C and observed for 14 days. Samples of mantle tissue were taken after the 14-day postexposure period and analysed for carbohydrate, lipid and protein. Three-way analysis of variance (ANOVA) did not reveal consistent trends in carbohydrate, lipid or protein concentrations due to sex of mussels, duration of emersion, air temperature or their interaction terms that indicated biological compensation to stress. Overall mean carbohydrate concentrations were greatest (range 447-615 mg/g dry wt) among the species, followed by protein (179-289 mg/g dry wt) and lipids (26.7-38.1 mg/g dry wt). These results have positive implications for conducting conservation activities, because emersion over the range of temperatures (15-35??C) and durations (15-60 min) examined did not appear acutely harmful to mussels.

Gaseous emissions from plants in controlled environments

NASA Technical Reports Server (NTRS)

Dubay, Denis T.

1988-01-01

Plant growth in a controlled ecological life support system may entail the build-up over extended time periods of phytotoxic concentrations of volatile organic compounds produced by the plants themselves. Ethylene is a prominent gaseous emission of plants, and is the focus of this report. The objective was to determine the rate of ethylene release by spring wheat, white potato, and lettuce during early, middle, and late growth stages, and during both the light and dark segments of the diurnal cycle. Plants grown hydroponically using the nutrient film technique were covered with plexiglass containers for 4 to 6 h. At intervals after enclosure, gas samples were withdrawn with a syringe and analyzed for ethylene with a gas chromatograph. Lettuce produced 10 to 100 times more ethylene than wheat or potato, with production rates ranging from 141 to 158 ng g-dry/wt/h. Wheat produced from 1.7 to 14.3 ng g-dry/wt/h, with senescent wheat producing the least amount and flowering wheat the most. Potatoes produced the least amount of ethylene, with values never exceeding 5 ng g-dry/wt/h. Lettuce and potatoes each produced ethylene at similar rates whether in dark period or light period. Ethylene sequestering of 33 to 43 percent by the plexiglass enclosures indicated that these production estimates may be low by one-third to one-half. These results suggest that concern for ethylene build-up in a contained atmosphere should be greatest when growing lettuce, and less when growing wheat or potato.

Differences in Establishment of Persistence of Vaccine and Wild Type Rubella Viruses in Fetal Endothelial Cells

PubMed Central

Perelygina, Ludmila; Adebayo, Adebola; Metcalfe, Maureen; Icenogle, Joseph

2015-01-01

Both wild type (WT) and vaccine rubella virus (RV) can pass through the placenta to infect a human fetus, but only wtRV routinely causes pathology. To investigate possible reasons for this, we compared establishment of persistence of wtRV and RA27/3 vaccine strains in fetal endothelial cells. We showed that yields of RA27/3 and wtRV were similar after the first round of replication, but then only vaccine-infected cultures went through a crisis characterized by partial cell loss and gradual decline of virus titer followed by recovery and establishment of persistent cultures with low levels of RA27/3 secretion. We compared various steps of virus replication, but we were unable to identify changes, which might explain the 2-log difference in RA27/3 and wtRV yields in persistently infected cultures. Whole genome sequencing did not reveal selection of virus variants in either the wtRV or RA27/3 cultures. Quantitative single-cell analysis of RV replication by in situ hybridization detected, on average, 1–4 copies of negative-strand RNA and ~50 copies of positive-strand genomic RNA in cells infected with both vaccine and WT viruses. The distinct characteristics of RA27/3 replication were the presence of large amounts of negative-strand RV RNA and RV dsRNA at the beginning of the crisis and the accumulation of high amounts of genomic RNA in a subpopulation of infected cells during crisis and persistence. These results suggest that RA27/3 can persist in fetal endothelial cells, but the characteristics of persistence and mechanisms for the establishment and maintenance of persistence are different from wtRV. PMID:26177032

Reconciling LCROSS and Orbital Neutron Water Abundance Estimates in Cabeus Crater

NASA Technical Reports Server (NTRS)

Elphic, Richard; Teodoro, Luis F.; Eke, Vincent R.; Paige, David A.; Siegler, Matthew A.; Colaprete, Anthony

2011-01-01

The Lunar Prospector Neutron Spectrometer (LPNS) first revealed Cabeus crater (84.9 deg S, 35.5degW) as having the highest inferred hydrogen on the Moon. Because of the broad LPNS footprint (approximately 40 km FWHM), the apparent peak water-equivalent hydrogen (WEH) concentration is only approximately 0.25 wt%, but could be much higher in smaller areas than the spectrometer footprint. Earlier image reconstruction work suggested that areas within permanent shadow have abundances approximately 1 wt% WEH. However, the LCROSS impact yielded total water estimates, ice plus vapor, of between 3 and 10 wt%. The large disagreement between LCROSS and apparent orbital values imply that either the ice is buried, by perhaps as much as 50 to 100 cm; or the ice distribution within Cabeus is spatially inhomogeneous, or both. Modeling reveals that the areal extent of a "shallow permafrost zone" is far greater than the area of permanent shadow. Ice can be virtually stable for billions of years within a few tens of centimeters of the surface in these areas. However, the LCROSS impact took place in an area of permanent shadow. If stably-trapped volatiles can be found in locales that receive occasional, oblique sunlight, landed missions may target these sites and eventual resource exploitation may be done more easily. Are orbital neutron data consistent with areally-extensive, volatile-rich cold traps? Orbital epithermal neutron data over the northern half of Cabeus (near the LCROSS impact site) are consistent with 0.2 wt% WEH or less in the "permafrost zone" near the crater. On the other hand, pixon reconstructions that confine the hydrogen enhancements to permanent shadow result in higher abundance estimates -- around 1 wt% if homogeneously mixed. But if the PSR abundance is increased to 10 wt%, consistent with the sum of all H-bearing compounds seen by LCROSS, a much larger-than-observed reduction in neutron count rate would be seen from orbit. It is likely that volatiles are inhomogeneously distributed, due to both impact processes and emplacement history. Two possibilities may bring consistency to the orbital and LCROSS measurements. Inhomogeneous lateral distribution: Consider the extreme case of a bimodal distribution within the crater -- dry and wet. In this case the epithermal leakage flux seen from orbit is a mixture of two different values, weighted according to fractional areas. Two possible outcomes, depending on whether the inferred leakage flux for the PSR or "permafrost" areas are considered. In the first case, approximately 40% of the PSR may be "wet", the remainder dry (and LCROSS was slightly lucky). However, if the whole area of permafrost is considered, then as little as 20% of the area will be as "wet" as the LCROSS results (and LCROSS was quite lucky). Inhomogeneous depth distribution: The leakage flux of thermal and epithermal neutrons depends on depth of burial of an icy layer beneath dry ferroan anorthosite soil (FAn). For the Cabeus PSR, the pixon reconstruction values for the epithermal flux allows a range of abundance and burial depth, while that of the thermal+epi detector constrains this range. (Uncertainties in iron abundance in the FAn can have significant impact on thermal neutron leakage flux estimates.) Between 20% and 40% of the Cabeus floor may be "wet", or alternatively a 5-10 wt% "wet" layer exists between 50 and 100 cm beneath a layer of dry regolith within the PSR. But volatile abundances of 5 wt% or more, distributed uniformly and homogeneously throughout the Cabeus PSR do not agree with orbital measurements

Hydrogen-Rich Syngas Production from Gasification and Pyrolysis of Solar Dried Sewage Sludge: Experimental and Modeling Investigations

PubMed Central

Ghrib, Amina; Friaa, Athar; Ouerghi, Aymen; Naoui, Slim; Belayouni, Habib

2017-01-01

Solar dried sewage sludge (SS) conversion by pyrolysis and gasification processes has been performed, separately, using two laboratory-scale reactors, a fixed-bed pyrolyzer and a downdraft gasifier, to produce mainly hydrogen-rich syngas. Prior to SS conversion, solar drying has been conducted in order to reduce moisture content (up to 10%). SS characterization reveals that these biosolids could be appropriate materials for gaseous products production. The released gases from SS pyrolysis and gasification present relatively high heating values (up to 9.96 MJ/kg for pyrolysis and 8.02  9.96 MJ/kg for gasification) due to their high contents of H2 (up to 11 and 7 wt%, resp.) and CH4 (up to 17 and 5 wt%, resp.). The yields of combustible gases (H2 and CH4) show further increase with pyrolysis. Stoichiometric models of both pyrolysis and gasification reactions were determined based on the global biomass formula, CαHβOγNδSε, in order to assist in the products yields optimization. PMID:28856162

Thermal Conductivity of Diamond Packed Electrospun PAN-Based Carbon Fibers Incorporated with Multi Wall Carbon Nanotubes.

PubMed

Dong, Qi; Lu, Chunyuan; Tulugan, Kelimu; Jin, Chunzi; Yoon, Soo Jong; Park, Yeong Min; Kim, Tae Gyu

2016-02-01

Multi wall carbon nanotubes (MWCNTs) and diamond are renowned as superlative material due to their relatively high thermal conductivity and hardness while comparing with any bulk materials. In this research, polyacrylonitrile (PAN) solution incorporated with MWCNTs at an alteration of mass fractions (0 wt%, 0.6 wt%, 1 wt%, 2 wt%) were fabricated via electrospinning under optimized parameters. Dried composite nanofibers were stabilized and carbonized, after which water base polytrafluorethylene (PTFE) mixed with nano diamond powder solution was spin coated on them. Scanning electron microscopy, Raman spectroscopy, X-ray scattering and Laserflash thermal conductivity were used to characterize the composite nanofiber sheets. The result shows that the thermal conductivity increased to 4.825 W/m K from 2.061 W/mK. The improvement of thermal conductivities is suggesting the incorporation of MWCNTs.

Wt-p53 action in human leukaemia cell lines corresponding to different stages of differentiation.

PubMed

Rizzo, M G; Zepparoni, A; Cristofanelli, B; Scardigli, R; Crescenzi, M; Blandino, G; Giuliacci, S; Ferrari, S; Soddu, S; Sacchi, A

1998-05-01

Recent studies support the potential application of the wt-p53 gene in cancer therapy. Expression of exogenous wt-p53 suppresses a variety of leukaemia phenotypes by acting on cell survival, proliferation and/or differentiation. As for tumour gene therapy, the final fate of the neoplastic cells is one of the most relevant points. We examined the effects of exogenous wt-p53 gene expression in several leukaemia cell lines to identify p53-responsive leukaemia. The temperature-sensitive p53Val135 mutant or the human wt-p53 cDNA was transduced in leukaemia cell lines representative of different acute leukaemia FAB subtypes, including M1 (KG1), M2 (HL-60), M3 (NB4), M5 (U937) and M6 (HEL 92.1.7), as well as blast crisis of chronic myelogenous leukaemia (BC-CML: K562, BV173) showing diverse differentiation features. By morphological, molecular and biochemical analyses, we have shown that exogenous wt-p53 gene expression induces apoptosis only in cells corresponding to M1, M2 and M3 of the FAB classification and in BC-CML showing morphological and cytochemical features of undifferentiated blast cells. In contrast, it promotes differentiation in the others. Interestingly, cell responsiveness was independent of the vector used and the status of the endogenous p53 gene.

Aberrant p63 and WT-1 expression in myoepithelial cells of pregnancy-associated breast cancer: implications for tumor aggressiveness and invasiveness

PubMed Central

Xu, Zheli; Wang, Wan; Deng, Chu-Xia; Man, Yan-gao

2009-01-01

Our recent studies revealed that focal alterations in breast myoepithelial cell layers significantly impact the biological presentation of associated epithelial cells. As pregnancy-associated breast cancer (PABC) has a significantly more aggressive clinical course and mortality rate than other forms of breast malignancies, our current study compared tumor suppressor expression in myoepithelial cells of PABC and non-PABC, to determine whether myoepithelial cells of PABC may have aberrant expression of tumor suppressors. Tissue sections from 20 cases of PABC and 20 cases of stage, grade, and age matched non-PABC were subjected to immunohistochemistry, and the expression of tumor suppressor maspin, p63, and Wilms' tumor 1 (WT-1) in calponin positive myoepithelial cells were statistically compared. The expression profiles of maspin, p63, and WT-1 in myoepithelial cells of all ducts encountered were similar between PABC and non-PABC. PABC, however, displayed several unique alterations in terminal duct and lobular units (TDLU), acini, and associated tumor tissues that were not seen in those of non-PABC, which included the absence of p63 and WT-1 expression in a vast majority of the myoepithelial cells, cytoplasmic localization of p63 in the entire epithelial cell population of some lobules, and substantially increasing WT-1 expression in vascular structures of the invasive cancer component. All or nearly all epithelial cells with aberrant p63 and WT-1 expression lacked the expression of estrogen receptor and progesterone receptor, whereas they had a substantially higher proliferation index than their counterparts with p63 and WT-1 expression. Hyperplastic cells with cytoplasmic p63 expression often adjacent to, and share a similar immunohistochemical and cytological profile with, invasive cancer cells. To our best knowledge, our main finings have not been previously reported. Our findings suggest that the functional status of myoepithelial cells may be significantly associated with tumor aggressiveness and invasiveness. PMID:19173015

Use of SLAM and PVRL4 and identification of pro-HB-EGF as cell entry receptors for wild type phocine distemper virus.

PubMed

Melia, Mary M; Earle, John Philip; Abdullah, Haniah; Reaney, Katherine; Tangy, Frederic; Cosby, Sara Louise

2014-01-01

Signalling lymphocyte activation molecule (SLAM) has been identified as an immune cell receptor for the morbilliviruses, measles (MV), canine distemper (CDV), rinderpest and peste des petits ruminants (PPRV) viruses, while CD46 is a receptor for vaccine strains of MV. More recently poliovirus like receptor 4 (PVRL4), also known as nectin 4, has been identified as a receptor for MV, CDV and PPRV on the basolateral surface of polarised epithelial cells. PVRL4 is also up-regulated by MV in human brain endothelial cells. Utilisation of PVRL4 as a receptor by phocine distemper virus (PDV) remains to be demonstrated as well as confirmation of use of SLAM. We have observed that unlike wild type (wt) MV or wtCDV, wtPDV strains replicate in African green monkey kidney Vero cells without prior adaptation, suggesting the use of a further receptor. We therefore examined candidate molecules, glycosaminoglycans (GAG) and the tetraspan proteins, integrin β and the membrane bound form of heparin binding epithelial growth factor (proHB-EGF),for receptor usage by wtPDV in Vero cells. We show that wtPDV replicates in Chinese hamster ovary (CHO) cells expressing SLAM and PVRL4. Similar wtPDV titres are produced in Vero and VeroSLAM cells but more limited fusion occurs in the latter. Infection of Vero cells was not inhibited by anti-CD46 antibody. Removal/disruption of GAG decreased fusion but not the titre of virus. Treatment with anti-integrin β antibody increased rather than decreased infection of Vero cells by wtPDV. However, infection was inhibited by antibody to HB-EGF and the virus replicated in CHO-proHB-EGF cells, indicating use of this molecule as a receptor. Common use of SLAM and PVRL4 by morbilliviruses increases the possibility of cross-species infection. Lack of a requirement for wtPDV adaptation to Vero cells raises the possibility of usage of proHB-EGF as a receptor in vivo but requires further investigation.

Extremely High Expression of Antisense RNA for Wilms' Tumor 1 in Active Osteoclasts: Suppression of Wilms' Tumor 1 Protein Expression during Osteoclastogenesis.

PubMed

Li, Yin-Ji; Kukita, Akiko; Kyumoto-Nakamura, Yukari; Kukita, Toshio

2016-09-01

Wilms' tumor 1 (WT1), a zinc-finger transcription regulator of the early growth response family, identified as the product of a tumor suppressor gene of Wilms' tumors, bears potential ability to induce macrophage differentiation in blood cell differentiation. Herein, we examined the involvement of WT1 in the regulation of osteoclastogenesis. We detected a high level of WT1 protein expression in osteoclast precursors; however, WT1 expression was markedly suppressed during osteoclastogenesis. We examined expression of WT1 transcripts in bone tissue by RNA in situ hybridization. We found a high level of antisense transcripts in osteoclasts actively resorbing bone in mandible of newborn rats. Expression of antisense WT1 RNA in mandible was also confirmed by Northern blot analysis and strand-specific RT-PCR. Overexpression of antisense WT1 RNA in RAW-D cells, an osteoclast precursor cell line, resulted in a marked enhancement of osteoclastogenesis, suggesting that antisense WT1 RNA functions to suppress expression of WT1 protein in osteoclastogenesis. High level expression of antisense WT1 RNA may contribute to commitment to osteoclastogenesis, and may allow osteoclasts to maintain or stabilize their differentiation state. Copyright © 2016 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.

Effect of intramammary infusion of chitosan hydrogels at drying-off on bovine mammary gland involution.

PubMed

Lanctôt, S; Fustier, P; Taherian, A R; Bisakowski, B; Zhao, X; Lacasse, P

2017-03-01

The transition from lactation to the dry period in dairy cows is a period of high risk for acquiring new intramammary infections. This risk is reduced when the involution of the mammary gland is completed. Accordingly, approaches that speed up the involution process after drying-off could reduce the incidence of mastitis. The current study aimed to develop a biological response modifier that could be injected into cow teats to promote immune cell migration and speed up involution. Chitosan, a natural polysaccharide derived from chitin, is able to trigger host innate immunity. We developed 2 formulations made from either high- or low-viscosity chitosan. Both are liquid at room temperature but form a hydrogel at body temperature. In the first experiment, each udder quarter of 7 Holstein cows in late lactation was randomly assigned at drying-off to receive one of the following intramammary infusions: 2.5 or 5 mL of 5% (wt/vol) low-viscosity chitosan hydrogel, 5 mL of 5% high-viscosity chitosan hydrogel, or 5 mL of water. Milk (mammary secretion) samples were collected from each quarter on d -4, -1 (drying-off), 1, 3, 5, 7, and 10. Milk somatic cell counts and the concentrations of involution markers such as BSA, lactate dehydrogenase, and lactoferrin were measured in each sample. In comparison with the control, the chitosan hydrogel infusions significantly hastened the increases in somatic cell counts, BSA and lactoferrin concentrations, and lactate dehydrogenase activity in mammary secretions. No major differences between sources or volumes of chitosan were observed for the measured parameters. The compatibility of this approach with an internal teat sealant was verified in the second experiment. Each udder quarter of 8 Holstein cows was randomly assigned at drying-off to receive one of the following intramammary infusions: 5 mL of 2% low-viscosity chitosan hydrogel, 4 g of an internal teat sealant, a combination of sealant and chitosan, or 5 mL of water. Milk (mammary secretion) samples were collected from each quarter on d -4, -1 (drying-off), 5, and 10 to measure involution markers. These results suggest that chitosan hydrogel infusion hastened mammary gland involution and activate immune response, which may reduce the risk of acquiring new intramammary infections during the drying-off period. Those results were not affected by the presence of the teat sealant, showing that both approaches are fully compatible and could be used in combination. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Preparation and combustion of coal-water fuel from the Sin Pun coal deposit, southern Thailand

DOE Office of Scientific and Technical Information (OSTI.GOV)

NONE

In response to an inquiry by the Department of Mineral Resources in Thailand, the Energy & Environmental Research Center (EERC) prepared a program to assess the responsiveness of Sin Pun lignite to the temperature and pressure conditions of hot-water drying. The results indicate that drying made several improvements in the coal, notably increases in heating value and carbon content and reductions in equilibrium moisture and oxygen content. The equilibrium moisture content decreased from 27 wt% for the raw coal to about 15 wt% for the hot-water-dried (HWD) coals. The energy density for a pumpable coal-water fuel (CWF) indicates an increasemore » from 4500 to 6100 Btu/lb by hot-water drying. Approximately 650 lb of HWD Sin Pun CWF were fired in the EERC`s combustion test facility. The fuel burned extremely well, with no feed problems noted during the course of the test. Fouling and slagging deposits each indicated a very low rate of ash deposition, with only a dusty layer formed on the cooled metal surfaces. The combustor was operated at between 20% and 25% excess air, resulting in a flue gas SO{sub 2} concentration averaging approximately 6500 parts per million.« less

Graphene oxide/alginate beads as adsorbents: Influence of the load and the drying method on their physicochemical-mechanical properties and adsorptive performance.

PubMed

Platero, Emiliano; Fernandez, Maria Emilia; Bonelli, Pablo Ricardo; Cukierman, Ana Lea

2017-04-01

Graphene oxide/alginate beads were prepared from lab-synthesized graphene oxide, varying its content within the beads (0.05, 0.125, and 0.25wt.%). Ethanol-drying and lyophilization were compared as drying methods to obtain suitable adsorbents which were later tested to the removal of a model organic molecule (methylene blue). The morphological and textural properties of all the beads were characterized by scanning electron microscopy and N 2 adsorption/desorption isotherms at -196°C, respectively. Limited porosity was obtained for all cases (S BET <60m 2 /g). Uniaxial compression tests were performed to assess the mechanical properties of the beads. Ethanol-dried ones exhibited higher Young's elasticity modulus (E=192kPa) than the lyophilized samples (twice at 0.25wt.% graphene oxide loading), which disclosed breakage points at lower deformation percentages. Adsorption experiments were conducted and dye adsorption isotherms were obtained for the beads with the best removal performance. The experimental data were better fitted by the Langmuir model. The highest maximum adsorption capacity (4.25mmol/g) was obtained for the lyophilized beads with the highest graphene oxide content. Mechanical properties were found to be affected also by the dye adsorption. Copyright © 2016 Elsevier Inc. All rights reserved.

Difference in the distribution and speciation of cellular nickel between nickel-tolerant and non-tolerant Nicotiana tabacum L. cv. BY-2 cells.

PubMed

Saito, Akihiro; Saito, Misa; Ichikawa, Yusuke; Yoshiba, Masaaki; Tadano, Toshiaki; Miwa, Eitaro; Higuchi, Kyoko

2010-02-01

To evaluate Ni dynamics at the subcellular level, the distribution and speciation of Ni were determined in wild-type (WT) and Ni-tolerant (NIT) tobacco BY-2 cell lines. When exposed to low but toxic levels of Ni, NIT cells were found to contain 2.5-fold more Ni (14% of whole-cell Ni values) in their cell walls than WT cells (6% of whole-cell Ni values). In addition to higher levels of Ni in the apoplast, a higher proportion (94%) of symplastic Ni was localized in the vacuoles of NIT cells than in the vacuoles of WT cells (81%). The concentration of cytosolic Ni in the NIT cells was significantly lower (18 nmol g(-1) FW) than that in the WT cells (85 nmol g(-1) FW). In silico simulation showed that 95% of vacuolar Ni was in the form of Ni-citrate complexes, and that free Ni(2+) was virtually absent in the NIT cells. On the other hand, the amount of free metal ions was markedly increased in WT cells because free citrate was depleted by chelation of Ni. A protoplast viability assay using BCECF-AM further demonstrated that the main mechanism that confers strong Ni tolerance was present in the symplast as opposed to the cell wall.

Coexpression of an unusual form of the EWS-WT1 fusion transcript and interleukin 2/15 receptor betamRNA in a desmoplastic small round cell tumour.

PubMed

Nakanishi, Y; Oinuma, T; Sano, M; Fuchinoue, F; Komatsu, K; Seki, T; Obana, Y; Tabata, M; Kikuchi, K; Shimamura, M; Ohmori, K; Nemoto, N

2006-10-01

The beta chain of the interleukin 2/15 receptor (IL-2/15Rbeta) is induced by the expression of the EWS-WT1. A case of desmoplastic small round cell tumour (DSRCT) expressing only an unusual EWS-WT1 treated by us is reported here. To characterise an unusual form of EWS-WT1. Frozen tissue sections of the axillary tumour were examined using a laser-assisted microdissection technique and reverse transcriptase polymerase chain reaction. The novel fusion of exon 8 of EWS and the defective exon 10 of WT1 (-KTS) was detected. Although it was an unusual form, the coexpression of the present EWS-WT1, IL-2/15Rbeta and Janus kinase (JAK1) mRNA was detected in the tumour cells. IL-2 and signal transducers and activators of transcription (STAT5) mRNA were detected in both tumour and stromal cells. The induction of the IL-2/15 receptor signalling pathway may contribute to tumorigenesis in DSRCT through a paracrine or an autocrine system, even though the EWS-WT1 was an unusual form.

Suitability of differently formulated dry powder Newcastle disease vaccines for mass vaccination of poultry.

PubMed

Huyge, Katrien; Van Reeth, Kristien; De Beer, Thomas; Landman, Wil J M; van Eck, Jo H H; Remon, Jean Paul; Vervaet, Chris

2012-04-01

Dry powders containing a live-attenuated Newcastle disease vaccine (LZ58 strain) and intended for mass vaccination of poultry were prepared by spray drying using mannitol in combination with trehalose or inositol, polyvinylpyrrolidone (PVP) and/or bovine serum albumin (BSA) as stabilizers. These powders were evaluated for vaccine stabilizing capacity during production and storage (at 6 °C and 25 °C), moisture content, hygroscopicity and dry powder dispersibility. A mixture design, varying the ratio of mannitol, inositol and BSA, was used to select the stabilizer combination which resulted in the desired powder properties (i.e. good vaccine stability during production and storage, low moisture content and hygroscopicity and good dry dispersibility). Inositol-containing powders had the same vaccine stabilizing capacity as trehalose powders, but were less hygroscopic. Incorporation of BSA enhanced the vaccine stability in the powders compared to PVP-containing formulations. However, increasing the BSA concentration increased the hygroscopicity and reduced the dry dispersibility of the powder. No valid mathematical model could be calculated for vaccine stability during production or storage, but the individual experiments indicated that a formulation combining mannitol, inositol and BSA in a ratio of 73.3:13.3:13.3 (wt/wt) resulted in the lowest vaccine titre loss during production (1.6-2.0 log(10) 50% egg infectious dose (EID(50)) and storage at 6 °C (max. 0.8 log(10) EID(50) after 6 months) in combination with a low moisture content (1.1-1.4%), low hygroscopicity (1.9-2.1% water uptake at 60% relative humidity) and good dry dispersibility properties. Copyright © 2011 Elsevier B.V. All rights reserved.

Combined mutagenesis of Rhodosporidium toruloides for improved production of carotenoids and lipids.

PubMed

Zhang, Chaolei; Shen, Hongwei; Zhang, Xibin; Yu, Xue; Wang, Han; Xiao, Shan; Wang, Jihui; Zhao, Zongbao K

2016-10-01

To improve production of lipids and carotenoids by the oleaginous yeast Rhodosporidium toruloides by screening mutant strains. Upon physical mutagenesis of the haploid strain R. toruloides np11 with an atmospheric and room temperature plasma method followed by chemical mutagenesis with nitrosoguanidine, a mutant strain, R. toruloides XR-2, formed dark-red colonies on a screening plate. When cultivated in nitrogen-limited media, XR-2 cells grew slower but accumulated 0.23 g lipids/g cell dry wt and 0.75 mg carotenoids/g CDW. To improve its production capacity, different amino acids and vitamins were supplemented. p-Aminobenzoic acid and tryptophan had beneficial effects on cell growth. When cultivated in nitrogen-limited media in the presence of selected vitamins, XR-2 accumulated 0.41 g lipids/g CDW and 0.69 mg carotenoids/g CDW. A mutant R. toruloides strain with improved production profiles for lipids and carotenoids was obtained, indicating its potential to use combined mutagenesis for a more productive phenotype.

Early detection of protozoan grazers in algal biofuel cultures.

PubMed

Day, John G; Thomas, Naomi J; Achilles-Day, Undine E M; Leakey, Raymond J G

2012-06-01

Future micro-algal biofuels will most likely be derived from open-pond production systems. These are by definition open to "invasion" by grazers, which could devastate micro-algal mass-cultures. There is an urgent requirement for methodologies capable of early detection and control of grazers in dense algal cultures. In this study a model system employing the marine alga Nannochloropsis oculata was challenged by grazers including ciliates, amoebae and a heterotrophic dinoflagellate. A FlowCAM flow-cytometer was used to detect all grazers investigated (size range <20->80 μm in length) in the presence of algae. Detection limits were <10 cells ml(-1) for both "large" and "small" model grazers, Euplotes vannus (80 × 45 μm) and an unidentified holotrichous ciliate (~18 × 8 μm) respectively. Furthermore, the system can distinguish the presence of ciliates in N. oculata cultures with biotechnologically relevant cell densities; i.e. >1.4 × 10(8) cells ml(-1) (>0.5 g l(-1) dry wt.). Copyright © 2012 Elsevier Ltd. All rights reserved.

The Role of Sub- and Supercritical CO2 as "Processing Solvent" for the Recycling and Sample Preparation of Lithium Ion Battery Electrolytes.

PubMed

Nowak, Sascha; Winter, Martin

2017-03-06

Quantitative electrolyte extraction from lithium ion batteries (LIB) is of great interest for recycling processes. Following the generally valid EU legal guidelines for the recycling of batteries, 50 wt % of a LIB cell has to be recovered, which cannot be achieved without the electrolyte; hence, the electrolyte represents a target component for the recycling of LIBs. Additionally, fluoride or fluorinated compounds, as inevitably present in LIB electrolytes, can hamper or even damage recycling processes in industry and have to be removed from the solid LIB parts, as well. Finally, extraction is a necessary tool for LIB electrolyte aging analysis as well as for post-mortem investigations in general, because a qualitative overview can already be achieved after a few minutes of extraction for well-aged, apparently "dry" LIB cells, where the electrolyte is deeply penetrated or even gellified in the solid battery materials.

Murine Denys-Drash syndrome: evidence of podocyte de-differentiation and systemic mediation of glomerulosclerosis.

PubMed

Patek, Charles E; Fleming, Stewart; Miles, Colin G; Bellamy, Christopher O; Ladomery, Michael; Spraggon, Lee; Mullins, John; Hastie, Nicholas D; Hooper, Martin L

2003-09-15

Denys-Drash syndrome (DDS) is caused by dominant mutations of the Wilms' tumour suppressor gene, WT1, and characterized by a nephropathy involving diffuse mesangial sclerosis, male pseudohermaphroditism and/or Wilms' tumourigenesis. Previously, we reported that heterozygosity for the Wt1tmT396 mutation induces DDS in heterozygous and chimeric (Wt1tmT396/+<-->+/+) mice. In the present study, the fate of Wt1 mutant cells in chimeric kidneys was assessed by in situ marker analysis, and immunocytochemistry was used to re-examine the claim that glomerulosclerosis (GS) is caused by loss of WT1 and persistent Pax-2 expression by podocytes. Wt1 mutant cells colonized glomeruli efficiently, including podocytes, but some sclerotic glomeruli contained no detectable Wt1 mutant cells. The development of GS was preceded by widespread loss of ZO-1 signal in podocytes (even in kidneys where <5% of glomeruli contained Wt1 mutant podocytes), increased intra-renal renin expression, and de novo podocyte TGF-beta1 expression, but not podocyte Pax-2 expression or loss of WT1, synaptopodin, alpha-actinin-4 or nephrin expression. However, podocytes in partially sclerotic glomeruli that still expressed WT1 at high levels showed reduced vimentin expression, cell cycle re-entry, and re-expressed desmin, cytokeratin and Pax-2. The results suggest that: (i) GS is not due to loss of WT1 expression by podocytes; (ii) podocyte Pax-2 expression reflects re-expression rather than persistent expression, and is the consequence of GS; (iii) GS is mediated systemically and the mechanism involves activation of the renin-angiotensin system; and (iv) podocytes undergo typical maturational changes but subsequently de-differentiate and revert to an immature phenotype during disease progression.

Wilms' tumor 1 (Wt1) regulates pleural mesothelial cell plasticity and transition into myofibroblasts in idiopathic pulmonary fibrosis

PubMed Central

Karki, Suman; Surolia, Ranu; Hock, Thomas David; Guroji, Purusotham; Zolak, Jason S.; Duggal, Ryan; Ye, Tong; Thannickal, Victor J.; Antony, Veena B.

2014-01-01

Pleural mesothelial cells (PMCs), which are derived from the mesoderm, exhibit an extraordinary capacity to undergo phenotypic changes during development and disease. PMC transformation and trafficking has a newly defined role in idiopathic pulmonary fibrosis (IPF); however, the contribution of Wilms' tumor 1 (Wt1)-positive PMCs to the generation of pathognomonic myofibroblasts remains unclear. PMCs were obtained from IPF lung explants and healthy donor lungs that were not used for transplantation. Short hairpin Wt1-knockdown PMCs (sh Wt1) were generated with Wt1 shRNA, and morphologic and functional assays were performed in vitro. Loss of Wt1 abrogated the PMC phenotype and showed evidence of mesothelial-to-mesenchymal transition (MMT), with a reduced expression of E-cadherin and an increase in the profibrotic markers α-smooth muscle actin (α-SMA) and fibronectin, along with increased migration and contractility, compared with that of the control. Migration of PMCs in response to active transforming growth factor (TGF)-β1 was assessed by live-cell imaging with 2-photon microscopy and 3D imaging, of Wt1-EGFP transgenic mice. Lineage-tracing experiments to map the fate of Wt1+ PMCs in mouse lung in response to TGF-β1 were also performed by using a Cre-loxP system. Our results, for the first time, demonstrate that Wt1 is necessary for the morphologic integrity of pleural membrane and that loss of Wt1 contributes to IPF via MMT of PMCs into a myofibroblast phenotype.—Karki, S., Surolia, R., Hock, T. D., Guroji, P., Zolak, J. S., Duggal, R., Ye, T., Thannickal, V., J., Antony, V. B. Wilms' tumor 1 (Wt1) regulates pleural mesothelial cell plasticity and transition into myofibroblasts in idiopathic pulmonary fibrosis. PMID:24265486

Renal ischemia-reperfusion injury and adenosine 2A receptor-mediated tissue protection: the role of CD4+ T cells and IFN-gamma.

PubMed

Day, Yuan-Ji; Huang, Liping; Ye, Hong; Li, Li; Linden, Joel; Okusa, Mark D

2006-03-01

A(2A) adenosine receptor (A(2A)R)-expressing bone marrow (BM)-derived cells contribute to the renal protective effect of A(2A) agonists in renal ischemia-reperfusion injury (IRI). We performed IRI in mice lacking T and B cells to determine whether A(2A)R expressed in CD4+ cells mediate protection from IRI. Rag-1 knockout (KO) mice were protected in comparison to wild-type (WT) mice when subjected to IRI. ATL146e, a selective A(2A) agonist, did not confer additional protection. IFN-gamma is an important early signal in IRI and is thought to contribute to reperfusion injury. Because IFN-gamma is produced by kidney cells and T cells we performed IRI in BM chimeras in which the BM of WT mice was reconstituted with BM from IFN-gamma KO mice (IFN-gamma KO-->WT chimera). We observed marked reduction in IRI in comparison to WT-->WT chimeras providing additional indirect support for the role of T cells. To confirm the role of CD4+ A(2A)R in mediating protection from IRI, Rag-1 KO mice were subjected to ischemia-reperfusion. The protection observed in Rag-1 KO mice was reversed in Rag-1 KO mice that were adoptively transferred WT CD4+ cells (WT CD4+-->Rag-1 KO) or A(2A) KO CD4+ cells (A(2A) KO CD4+-->Rag-1 KO). ATL146e reduced injury in WT CD4+-->Rag-1 KO mice but not in A(2A) KO CD4+-->Rag-1 KO mice. Rag-1 KO mice reconstituted with CD4+ cells derived from IFN-gamma KO mice (IFN-gamma CD4+-->Rag-1 KO) were protected from IRI; ATL146e conferred no additional protection. These studies demonstrate that CD4+ IFN-gamma contributes to IRI and that A(2A) agonists mediate protection from IRI through action on CD4+ cells.

Occurrence of tributyltin (TBT)-resistant bacteria is not related to TBT pollution in Mekong River and coastal sediment: with a hypothesis of selective pressure from suspended solid.

PubMed

Suehiro, Fujiyo; Mochizuki, Hiroko; Nakamura, Shinji; Iwata, Hisato; Kobayashi, Takeshi; Tanabe, Shinsuke; Fujimori, Yoshifumi; Nishimura, Fumitake; Tuyen, Bui Cach; Tana, Touch Seang; Suzuki, Satoru

2007-07-01

Tributyltin (TBT) is organotin compound that is toxic to aquatic life ranging from bacteria to mammals. This study examined the concentration of TBT in sediment from and near the Mekong River and the distribution of TBT-resistant bacteria. TBT concentrations ranged from <2.4 to 2.4 ng/g (dry wt) in river sediment and <2.4-15 ng g(-1) (dry wt) in harbor sediment. Viable count of total bacteria ranged from 2.0 x 10(4) to 1.4 x 10(7)cfu/g, and counts of TBT-resistant bacteria ranged <1.0 x 10(2) to 2.5 x 10(4)cfu/g. The estimated occurrence rate of TBT-resistant bacteria ranged from <0.01 to 34% and was highest in upstream sites in Cambodia. The occurrences of TBT in the sediment and of TBT-resistant bacteria were unrelated, and chemicals other than TBT might induce TBT resistance. TBT-resistant bacteria were more abundant in the dry season than in the rainy season. Differences in the selection process of TBT-resistant bacteria between dry and rainy seasons were examined using an advection-diffusion model of a suspended solid (SS) that conveys chemicals. The estimated dilution-diffusion time over a distance of 120 km downstream from a release site was 20 days during dry season and 5 days during rainy season, suggesting that bacteria at the sediment surface could be exposed to SS for longer periods during dry season.

Regulation of HtrA2 on WT1 gene expression under imatinib stimulation and its effects on the cell biology of K562 cells.

PubMed

Zhang, Lixia; Li, Yan; Li, Xiaoyan; Zhang, Qing; Qiu, Shaowei; Zhang, Qi; Wang, Min; Xing, Haiyan; Rao, Qing; Tian, Zheng; Tang, Kejing; Wang, Jianxiang; Mi, Yingchang

2017-09-01

The aim of the present study was to investigate the regulation of Wilms Tumor 1 (WT1) by serine protease high-temperature requirement protein A2 (HtrA2), a member of the Htr family, in K562 cells. In addition, the study aimed to observe the effect of this regulation on cell biological functions and its associated mechanisms. Expression of WT1 and HtrA2 mRNA, and proteins following imatinib and the HtrA2 inhibitor 5-[5-(2-nitrophenyl) furfuryl iodine]-1, 3-diphenyl-2-thiobarbituric acid (UCF-101) treatment was detected with reverse transcription-quantitative polymerase chain reaction and western blot analysis. Subsequent to treatment with drugs and UCF-101, the proliferative function of K562 cells was detected using MTT assays, and the rate of apoptosis was detected using Annexin V with propidium iodide flow cytometry in K562 cells. The protein levels in the signaling pathway were analyzed using western blotting following treatment with imatinib and UCF-101. In K562 cells, imatinib treatment activated HtrA2 gene at a transcription level, while the WT1 gene was simultaneously downregulated. Following HtrA2 inhibitor (UCF-101) treatment, the downregulation of WT1 increased gradually. At the protein level, imatinib induced the increase in HtrA2 protein level and concomitantly downregulated WT1 protein level. Subsequent to HtrA2 inhibition by UCF-101, the WT1 protein level decreased temporarily, but eventually increased. Imatinib induced apoptosis in K562 cells, but this effect was attenuated by the HtrA2 inhibitor UCF-101, resulting in the upregulation of the WT1 protein level. However; UCF-101 did not markedly change the proliferation inhibition caused by imatinib. Imatinib activated the p38 mitogen activated protein kinase (p38 MAPK) signaling pathway in K562 cells, and UCF-101 affected the activation of imatinib in the p38 MAPK signaling pathway. Imatinib inhibited the extracellular signal-related kinase (ERK1/2) pathway markedly and persistently, but UCF-101 exhibited no notable effect on the inhibition of the ERK1/2 pathway. HtrA2 and its regulatory effect on WT1 may affect the sensitivity of BCR/ABL(+) cell lines to target therapy drugs through different mechanisms. Regulation of WT1 by HtrA2 occurs in K562 cells, and the regulation may affect the apoptosis of K562 cells under the stress caused by chemotherapeutic treatment. The p38 MAPK signaling pathway, which serves an important role in cell apoptosis, is a downstream pathway of this regulation.

Secreted protein acidic and rich in cysteine functions in colitis via IL17A regulation in mucosal CD4+ T cells.

PubMed

Tanaka, Makoto; Takagi, Tomohisa; Naito, Yuji; Uchiyama, Kazuhiko; Hotta, Yuma; Toyokawa, Yuki; Ushiroda, Chihiro; Hirai, Yasuko; Aoi, Wataru; Higashimura, Yasuki; Mizushima, Katsura; Okayama, Tetsuya; Katada, Kazuhiro; Kamada, Kazuhiro; Ishikawa, Takeshi; Handa, Osamu; Itoh, Yoshito

2018-03-01

Secreted protein acidic and rich in cysteine (SPARC) is a matricellular glycol that regulates cell proliferation, tissue repair, and tumorigenesis. Despite evidence linking SPARC to inflammation, the mechanisms are unclear. Accordingly, the role of SPARC in intestinal inflammation was investigated. Colitis was induced in wild-type (WT) and SPARC knockout (KO) mice using trinitrobenzene sulfonic acid (TNBS). Colons were assessed for damage; leukocyte infiltration; Tnf, Ifng, Il17a, and Il10 mRNA expression; and histology. Cytokine profiling of colonic lamina propria mononuclear cells (LPMCs) was performed by flow cytometry. Naïve CD4 + T cells were isolated from WT and SPARC KO mouse spleens, and the effect of SPARC on Th17 cell differentiation was examined. Recombination activating gene 1 knockout (RAG1 KO) mice reconstituted with T cells from either WT or SPARC KO mice were investigated. Trinitrobenzene sulfonic acid exposure significantly reduced bodyweight and increased mucosal inflammation, leukocyte infiltration, and Il17a mRNA expression in WT relative to SPARC KO mice. The percentage of IL17A-producing CD4 + T cells among LPMCs from KO mice was lower than that in WT mice when both groups were exposed to TNBS. Th17 cell differentiation was suppressed in cells from SPARC KO mice. In the T cell transfer colitis model, RAG1 KO mice receiving T cells from WT mice were more severely affected than those reconstituted with cells from SPARC KO mice. Secreted protein acidic and rich in cysteine accelerates colonic mucosal inflammation via modulation of IL17A-producing CD4 + T cells. SPARC is a potential therapeutic target for conditions involving intestinal inflammation. © 2017 Journal of Gastroenterology and Hepatology Foundation and John Wiley & Sons Australia, Ltd.

PAX3 is expressed in the stromal compartment of the developing kidney and in Wilms tumors with myogenic phenotype.

PubMed

Hueber, Pierre-Alain; Fukuzawa, Ryuji; Elkares, Reyhan; Chu, Leelee; Blumentkrantz, Miriam; He, Shu-Jie; Anaka, Matthew R; Reeve, Anthony E; Eccles, Michael; Jabado, Nada; Iglesias, Diana M; Goodyer, Paul R

2009-01-01

Wilms tumor (WT) is the most frequent renal neoplasm of childhood; a myogenic component is observed in 5% to 10% of tumors. We demonstrate for the first time that myogenic WTs are associated with expression of PAX3, a transcription factor known to specify myoblast cell fate during muscle development. In a panel of 20 WTs, PAX3 was identified in 13 of 13 tumor samples with myogenic histopathology but was absent in 7 of 7 tumors lacking a myogenic component. Furthermore, we show that PAX3 is expressed in the metanephric mesenchyme and stromal compartment of developing mouse kidney. Modulation of endogenous PAX3 expression in human embryonic kidney (HEK293) cells influenced cell migration in in vitro assays. Mutations of WT1 were consistently associated with PAX3 expression in WTs, and modulation of WT1 expression in HEK293 cells was inversely correlated with the level of endogenous PAX3 protein. We demonstrate abundant PAX3 and absence of PAX2 expression in a novel cell line (WitP3) isolated from the stromal portion of a WT bearing a homozygous deletion of the WT1 gene. We hypothesize that PAX3 sets stromal cell fate in developing kidney but is normally suppressed by WT1 during the mesenchyme-to-epithelium transition leading to nephrogenesis. Loss of WT1 permits aberrant PAX3 expression in a subset of WTs with myogenic phenotype.

Mössbauer, EPR, and Modeling Study of Iron Trafficking and Regulation in Δccc1 and CCC1-up Saccharomyces cerevisiae

PubMed Central

2015-01-01

Strains lacking and overexpressing the vacuolar iron (Fe) importer CCC1 were characterized using Mössbauer and EPR spectroscopies. Vacuolar Fe import is impeded in Δccc1 cells and enhanced in CCC1-up cells, causing vacuolar Fe in these strains to decline and accumulate, respectively, relative to WT cells. Cytosolic Fe levels should behave oppositely. The Fe content of Δccc1 cells grown under low-Fe conditions was similar to that in WT cells. Most Fe was mitochondrial with some nonheme high spin (NHHS) FeII present. Δccc1 cells grown with increasing Fe concentration in the medium contained less total Fe, less vacuolar HS FeIII, and more NHHS FeII than in comparable WT cells. As the Fe concentration in the growth medium increased, the concentration of HS FeIII in Δccc1 cells increased to just 60% of WT levels, while NHHS FeII increased to twice WT levels, suggesting that the NHHS FeII was cytosolic. Δccc1 cells suffered more oxidative damage than WT cells, suggesting that the accumulated NHHS FeII promoted Fenton chemistry. The Fe concentration in CCC1-up cells was higher than in WT cells; the extra Fe was present as NHHS FeII and FeIII and as FeIII oxyhydroxide nanoparticles. These cells contained less mitochondrial Fe and exhibited less ROS damage than Δccc1 cells. CCC1-up cells were adenine-deficient on minimal medium; supplementing with adenine caused a decline of NHHS FeII suggesting that some of the NHHS FeII that accumulated in these cells was associated with adenine deficiency rather than the overexpression of CCC1. A mathematical model was developed that simulated changes in Fe distributions. Simulations suggested that only a modest proportion of the observed NHHS FeII in both strains was the cytosolic form of Fe that is sensed by the Fe import regulatory system. The remainder is probably generated by the reduction of the vacuolar NHHS FeIII species. PMID:24785783

Mössbauer, EPR, and modeling study of iron trafficking and regulation in Δccc1 and CCC1-up Saccharomyces cerevisiae.

PubMed

Cockrell, Allison; McCormick, Sean P; Moore, Michael J; Chakrabarti, Mrinmoy; Lindahl, Paul A

2014-05-13

Strains lacking and overexpressing the vacuolar iron (Fe) importer CCC1 were characterized using Mössbauer and EPR spectroscopies. Vacuolar Fe import is impeded in Δccc1 cells and enhanced in CCC1-up cells, causing vacuolar Fe in these strains to decline and accumulate, respectively, relative to WT cells. Cytosolic Fe levels should behave oppositely. The Fe content of Δccc1 cells grown under low-Fe conditions was similar to that in WT cells. Most Fe was mitochondrial with some nonheme high spin (NHHS) Fe(II) present. Δccc1 cells grown with increasing Fe concentration in the medium contained less total Fe, less vacuolar HS Fe(III), and more NHHS Fe(II) than in comparable WT cells. As the Fe concentration in the growth medium increased, the concentration of HS Fe(III) in Δccc1 cells increased to just 60% of WT levels, while NHHS Fe(II) increased to twice WT levels, suggesting that the NHHS Fe(II) was cytosolic. Δccc1 cells suffered more oxidative damage than WT cells, suggesting that the accumulated NHHS Fe(II) promoted Fenton chemistry. The Fe concentration in CCC1-up cells was higher than in WT cells; the extra Fe was present as NHHS Fe(II) and Fe(III) and as Fe(III) oxyhydroxide nanoparticles. These cells contained less mitochondrial Fe and exhibited less ROS damage than Δccc1 cells. CCC1-up cells were adenine-deficient on minimal medium; supplementing with adenine caused a decline of NHHS Fe(II) suggesting that some of the NHHS Fe(II) that accumulated in these cells was associated with adenine deficiency rather than the overexpression of CCC1. A mathematical model was developed that simulated changes in Fe distributions. Simulations suggested that only a modest proportion of the observed NHHS Fe(II) in both strains was the cytosolic form of Fe that is sensed by the Fe import regulatory system. The remainder is probably generated by the reduction of the vacuolar NHHS Fe(III) species.

Nanofibrous poly(lactide-co-glycolide) membranes loaded with diamond nanoparticles as promising substrates for bone tissue engineering

PubMed Central

Parizek, Martin; Douglas, Timothy EL; Novotna, Katarina; Kromka, Alexander; Brady, Mariea A; Renzing, Andrea; Voss, Eske; Jarosova, Marketa; Palatinus, Lukas; Tesarek, Pavel; Ryparova, Pavla; Lisa, Věra; dos Santos, Ana M; Bacakova, Lucie

2012-01-01

Background Nanofibrous scaffolds loaded with bioactive nanoparticles are promising materials for bone tissue engineering. Methods In this study, composite nanofibrous membranes containing a copolymer of L-lactide and glycolide (PLGA) and diamond nanoparticles were fabricated by an electrospinning technique. PLGA was dissolved in a mixture of methylene chloride and dimethyl formamide (2:3) at a concentration of 2.3 wt%, and nanodiamond (ND) powder was added at a concentration of 0.7 wt% (about 23 wt% in dry PLGA). Results In the composite scaffolds, the ND particles were either arranged like beads in the central part of the fibers or formed clusters protruding from the fibers. In the PLGA-ND membranes, the fibers were thicker (diameter 270 ± 9 nm) than in pure PLGA meshes (diameter 218 ± 4 nm), but the areas of pores among these fibers were smaller than in pure PLGA samples (0.46 ± 0.02 μm2 versus 1.28 ± 0.09 μm2 in pure PLGA samples). The PLGA-ND membranes showed higher mechanical resistance, as demonstrated by rupture tests of load and deflection of rupture probe at failure. Both types of membranes enabled the attachment, spreading, and subsequent proliferation of human osteoblast-like MG-63 cells to a similar extent, although these values were usually lower than on polystyrene dishes. Nevertheless, the cells on both types of membranes were polygonal or spindle-like in shape, and were distributed homogeneously on the samples. From days 1–7 after seeding, their number rose continuously, and at the end of the experiment, these cells were able to create a confluent layer. At the same time, the cell viability, evaluated by a LIVE/DEAD viability/cytotoxicity kit, ranged from 92% to 97% on both types of membranes. In addition, on PLGA-ND membranes, the cells formed well developed talin-containing focal adhesion plaques. As estimated by the determination of tumor necrosis factor-alpha levels in the culture medium and concentration of intercellular adhesion molecule-1, MG-63 cells, and RAW 264.7 macrophages on these membranes did not show considerable inflammatory activity. Conclusion This study shows that nanofibrous PLGA membranes loaded with diamond nanoparticles have interesting potential for use in bone tissue engineering. PMID:22619532

Nanofibrous poly(lactide-co-glycolide) membranes loaded with diamond nanoparticles as promising substrates for bone tissue engineering.

PubMed

Parizek, Martin; Douglas, Timothy E L; Novotna, Katarina; Kromka, Alexander; Brady, Mariea A; Renzing, Andrea; Voss, Eske; Jarosova, Marketa; Palatinus, Lukas; Tesarek, Pavel; Ryparova, Pavla; Lisa, Věra; dos Santos, Ana M; Warnke, Patrick H; Bacakova, Lucie

2012-01-01

Nanofibrous scaffolds loaded with bioactive nanoparticles are promising materials for bone tissue engineering. In this study, composite nanofibrous membranes containing a copolymer of L-lactide and glycolide (PLGA) and diamond nanoparticles were fabricated by an electrospinning technique. PLGA was dissolved in a mixture of methylene chloride and dimethyl formamide (2:3) at a concentration of 2.3 wt%, and nanodiamond (ND) powder was added at a concentration of 0.7 wt% (about 23 wt% in dry PLGA). In the composite scaffolds, the ND particles were either arranged like beads in the central part of the fibers or formed clusters protruding from the fibers. In the PLGA-ND membranes, the fibers were thicker (diameter 270 ± 9 nm) than in pure PLGA meshes (diameter 218 ± 4 nm), but the areas of pores among these fibers were smaller than in pure PLGA samples (0.46 ± 0.02 μm(2) versus 1.28 ± 0.09 μm(2) in pure PLGA samples). The PLGA-ND membranes showed higher mechanical resistance, as demonstrated by rupture tests of load and deflection of rupture probe at failure. Both types of membranes enabled the attachment, spreading, and subsequent proliferation of human osteoblast-like MG-63 cells to a similar extent, although these values were usually lower than on polystyrene dishes. Nevertheless, the cells on both types of membranes were polygonal or spindle-like in shape, and were distributed homogeneously on the samples. From days 1-7 after seeding, their number rose continuously, and at the end of the experiment, these cells were able to create a confluent layer. At the same time, the cell viability, evaluated by a LIVE/DEAD viability/cytotoxicity kit, ranged from 92% to 97% on both types of membranes. In addition, on PLGA-ND membranes, the cells formed well developed talin-containing focal adhesion plaques. As estimated by the determination of tumor necrosis factor-alpha levels in the culture medium and concentration of intercellular adhesion molecule-1, MG-63 cells, and RAW 264.7 macrophages on these membranes did not show considerable inflammatory activity. This study shows that nanofibrous PLGA membranes loaded with diamond nanoparticles have interesting potential for use in bone tissue engineering.

Mercury in the atmospheric and coastal environments of Mexico.

PubMed

Ruelas-Inzunza, Jorge; Delgado-Alvarez, Carolina; Frías-Espericueta, Martín; Páez-Osuna, Federico

2013-01-01

In Mexico, published studies relating to the occurrence of Hg in the environment are limited. Among the main sources of Hg in Mexico are mining and refining of Auand Hg, chloralkali plants, Cu smelting, residential combustion of wood, carbo electric plants, and oil refineries. Hg levels are highly variable in the atmospheric compartment because of the atmospheric dynamics and ongoing metal exchange with the terrestrial surface. In atmospheric studies, Hg levels are usually reported as total gaseous Hg (TGM). In Mexico, TGM values ranged from 1.32 ng m-3 in Hidalgo state (a rural agricultural area) to 71.82 ng m-3 in Zacatecas state (an area where brick manufacturers use mining wastes as a raw material).Published information on mercury levels in the coastal environment comprise 21 studies, representing 21 areas, in which sediments constituted the substrate that was analyzed for Hg. In addition, water samples were analyzed for Hg in nine studies.Few studies exist on Hg levels in the Caribbean and in the southwest of the country where tourism is rapidly increasing. Hence, there is a need for establishing baseline levels of mercury in these increasingly visited areas. In regions where studies have been undertaken, Hg levels in sediments were highly variable. Variations in Hg sediment levels mainly result from geological factors and the varying degree of anthropogenic impacts in the studied areas. In areas that still have pristine or nearly pristine environments (e.g., coast, Baja California, Todos Santos Bay, and La Paz lagoon), sediment Hg levels ranged from <0.006 to 0.35 j.lg g-1 on a dry wt basis.When higher levels exist (0.34-57.94 j.lg g-1 on a dry wt basis), the environment generally shows the influence of inputs from mining, oil processing, agriculture,geothermal events, or harmful algal bloom events (e.g., Guaymas Bay and Coatzacoalcos estuary). From chronological studies performed in selected coastal lagoons in NW Mexico, it is clear that Hg fluxes to sediments have increased from2- to 15-fold in recent years. Since the 1940s, historical increases of Hg fluxes have resulted from higher agricultural waste releases and exhaust from the thermo electric plants. The levels of Hg in water reveal a moderate to elevated contamination of some Mexican coastal sites. In Urias lagoon (NW Mexico), moderate to high levels were found in the dissolved and suspended fraction, and these are related to shipping, the fishing industry, domestic effluents, and the presence of a thermoelectric plant. In Coatzacoalcos (SE Mexico), extremely elevated Hg levels were found during the decade of the 1970s. Low to moderate levels of Hg were measured in waters from the Alvarado lagoon (SE Mexico); those concentrations appear to be associated with river waters that became enriched with organic matter and suspended solids inthe brackish mixing zone.Regarding the Hg content in invertebrates, the use of bivalves (oysters and mussels)as biomonitors must be established along the coastal zones of Mexico, because some coastal lagoons have not been previously monitored. In addition, more research is needed to investigate shrimp farms that are associated with agricultural basins and receive effluents from several anthropogenic sources (e.g., mining activity and urban discharges). Hg residues in several vertebrate groups collected in Mexico have been studied.These include mammals, birds, reptiles, and fish. In elasmobranch species, the highest Hg concentration (27.2 flg g-1 dry wt) was found in the muscle of the smooth hammer head shark (Sphyrna zygaena). Teleost fish are the vertebrate group that has been most studied, with regard to Hg residue content; the highest value (5.67 11g g-1dry wt) was detected in the striped marlin (T. audax). Among reptiles, only marine turtles were studied; Hg levels found ranged from 0.795 in the liver to 0.0006flg g-1dry wt in the blood of L. olivacea. In birds, the highest Hg concentration (5.08 flg g-1dry wt) detected was in the liver of the olivaceous cormorant (P. olivaceous).Specimens from stranded marine mammals were also analyzed; levels of Hg ranged from 70.35 flg g-1 dry wt in the liver of stranded spinner dolphin (S. longirostris ), to0.145 flg g-1 dry wt in the muscle of gray whale (E. robustus). The presence of Hgin these marine animals is not thought to have caused the stranding of the animals.Other organisms like macroalgae and vestimentiferan tube worms were used to monitor the occurrence of Hg in the aquatic environment; levels were comparable to data reported on similar organisms from other areas of the world. Few investigation shave been carried out concerning the mercury content in human organs/tissues in Mexico. Considering the potential deleterious effects of Hg on kidney, lung, and the central nervous system, more information about human exposure to organic and inorganic forms of mercury and their effects is needed, both in Mexico and elsewhere.

The Metallothionein-Null Phenotype Is Associated with Heightened Sensitivity to Lead Toxicity and an Inability to Form Inclusion Bodies

PubMed Central

Qu, Wei; Diwan, Bhalchandra A.; Liu, Jie; Goyer, Robert A.; Dawson, Tammy; Horton, John L.; Cherian, M. George; Waalkes, Michael P.

2002-01-01

Susceptibility to lead toxicity in MT-null mice and cells, lacking the major forms of the metallothionein (MT) gene, was compared to wild-type (WT) mice or cells. Male MT-null and WT mice received lead in the drinking water (0 to 4000 ppm) for 10 to 20 weeks. Lead did not alter body weight in any group. Unlike WT mice, lead-treated MT-null mice showed dose-related nephromegaly. In addition, after lead exposure renal function was significantly diminished in MT-null mice in comparison to WT mice. MT-null mice accumulated less renal lead than WT mice and did not form lead inclusion bodies, which were present in the kidneys of WT mice. In gene array analysis, renal glutathione S-transferases were up-regulated after lead in MT-null mice only. In vitro studies on fibroblast cell lines derived from MT-null and WT mice showed that MT-null cells were much more sensitive to lead cytotoxicity. MT-null cells accumulated less lead and formed no inclusion bodies. The MT-null phenotype seems to preclude lead-induced inclusion body formation and increases lead toxicity at the organ and cellular level despite reducing lead accumulation. This study reveals important roles for MT in chronic lead toxicity, lead accumulation, and inclusion body formation. PMID:11891201

Glyceraldehyde-3-phosphate Dehydrogenase (GAPDH) Aggregation Causes Mitochondrial Dysfunction during Oxidative Stress-induced Cell Death*

PubMed Central

Itakura, Masanori; Kubo, Takeya; Kaneshige, Akihiro; Harada, Naoki; Izawa, Takeshi; Azuma, Yasu-Taka; Kuwamura, Mitsuru; Yamaji, Ryouichi; Takeuchi, Tadayoshi

2017-01-01

Glycolytic glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a multifunctional protein that also mediates cell death under oxidative stress. We reported previously that the active-site cysteine (Cys-152) of GAPDH plays an essential role in oxidative stress-induced aggregation of GAPDH associated with cell death, and a C152A-GAPDH mutant rescues nitric oxide (NO)-induced cell death by interfering with the aggregation of wild type (WT)-GAPDH. However, the detailed mechanism underlying GAPDH aggregate-induced cell death remains elusive. Here we report that NO-induced GAPDH aggregation specifically causes mitochondrial dysfunction. First, we observed a correlation between NO-induced GAPDH aggregation and mitochondrial dysfunction, when GAPDH aggregation occurred at mitochondria in SH-SY5Y cells. In isolated mitochondria, aggregates of WT-GAPDH directly induced mitochondrial swelling and depolarization, whereas mixtures containing aggregates of C152A-GAPDH reduced mitochondrial dysfunction. Additionally, treatment with cyclosporin A improved WT-GAPDH aggregate-induced swelling and depolarization. In doxycycline-inducible SH-SY5Y cells, overexpression of WT-GAPDH augmented NO-induced mitochondrial dysfunction and increased mitochondrial GAPDH aggregation, whereas induced overexpression of C152A-GAPDH significantly suppressed mitochondrial impairment. Further, NO-induced cytochrome c release into the cytosol and nuclear translocation of apoptosis-inducing factor from mitochondria were both augmented in cells overexpressing WT-GAPDH but ameliorated in C152A-GAPDH-overexpressing cells. Interestingly, GAPDH aggregates induced necrotic cell death via a permeability transition pore (PTP) opening. The expression of either WT- or C152A-GAPDH did not affect other cell death pathways associated with protein aggregation, such as proteasome inhibition, gene expression induced by endoplasmic reticulum stress, or autophagy. Collectively, these results suggest that NO-induced GAPDH aggregation specifically induces mitochondrial dysfunction via PTP opening, leading to cell death. PMID:28167533

Saccharification of Cellulose by Recombinant Rhodococcus opacus PD630 Strains

PubMed Central

Hetzler, Stephan; Bröker, Daniel

2013-01-01

The noncellulolytic actinomycete Rhodococcus opacus strain PD630 is the model oleaginous prokaryote with regard to the accumulation and biosynthesis of lipids, which serve as carbon and energy storage compounds and can account for as much as 87% of the dry mass of the cell in this strain. In order to establish cellulose degradation in R. opacus PD630, we engineered strains that episomally expressed six different cellulase genes from Cellulomonas fimi ATCC 484 (cenABC, cex, cbhA) and Thermobifida fusca DSM43792 (cel6A), thereby enabling R. opacus PD630 to degrade cellulosic substrates to cellobiose. Of all the enzymes tested, five exhibited a cellulase activity toward carboxymethyl cellulose (CMC) and/or microcrystalline cellulose (MCC) as high as 0.313 ± 0.01 U · ml−1, but recombinant strains also hydrolyzed cotton, birch cellulose, copy paper, and wheat straw. Cocultivations of recombinant strains expressing different cellulase genes with MCC as the substrate were carried out to identify an appropriate set of cellulases for efficient hydrolysis of cellulose by R. opacus. Based on these experiments, the multicellulase gene expression plasmid pCellulose was constructed, which enabled R. opacus PD630 to hydrolyze as much as 9.3% ± 0.6% (wt/vol) of the cellulose provided. For the direct production of lipids from birch cellulose, a two-step cocultivation experiment was carried out. In the first step, 20% (wt/vol) of the substrate was hydrolyzed by recombinant strains expressing the whole set of cellulase genes. The second step was performed by a recombinant cellobiose-utilizing strain of R. opacus PD630, which accumulated 15.1% (wt/wt) fatty acids from the cellobiose formed in the first step. PMID:23793636

Mineralogy of the Hydrous Lower Mantle

NASA Astrophysics Data System (ADS)

Shim, S. H.; Chen, H.; Leinenweber, K. D.; Kunz, M.; Prakapenka, V.; Bechtel, H.; Liu, Z.

2017-12-01

The hydrous ringwoodite inclusions found in diamonds suggest water storage in the mantle transition zone. However, water storage in the lower mantle remains unclear. Bridgmanite and magnesiowustite appear to have very little storage capacity for water. Here, we report experimental results indicating significant changes in the lower-mantle mineralogy under the presence of water. We have synthesized Mg2SiO4 ringwoodite with 2 wt% water in multi-anvil press at 20 GPa and 1573 K at ASU. The hydrous ringwoodite sample was then loaded to diamond anvil cells with Ar or Ne as a pressure medium. We heated the pure hydrous ringwoodite samples at lower-mantle pressure using a CO2 laser heating system at ASU. We measured X-ray diffraction patterns at the GSECARS sector of the Advanced Photon Source (APS) and 12.2.2 sector of the Advanced Light Source (ALS). For the separate Pt-mixed samples, we have conducted in situ heating at the beamlines using near IR laser heating systems. We measured the infrared spectra of the heated samples at high pressure and after pressure quench at 1.4.4 sector of ALS. In the in situ experiments with hydrous ringwoodite + Pt mixture as a starting material, we found formation of stishovite together with bridgmanite and periclase during heating with a near IR laser beams at 1300-2500 K and 35-66 GPa. However, some hydrous ringwoodite still remains even after a total of 45 min of heating. In contrast, the hydrous ringwoodite samples heated without Pt by CO2 laser beams are transformed completely to bridgmanite, periclase and stishovite at 31-55 GPa and 1600-1900 K. We have detected IR active OH mode of stishovite from the samples heated at lower-mantle pressures. The unit-cell volume of stishovite measured after pressure quench is greater than that of dry stishovite by 0.3-0.6%, supporting 0.5-1 wt% of H2O in stishovite in these samples. Stishovite is a thermodynamically forbidden phase in the dry lower mantle because of the existence of periclase and bridgmanite. However, our results indicate that stishovite can exist together with periclase and bridgmanite when water is present, because water is stored in stishovite. Therefore, water-rich parts of the lower mantle, such as regions with subducting slabs, would have distinct mineralogy from their dry counterparts, containing stishovite as a water storage mineral.

Mutation in the Fas Pathway Impairs CD8+ T Cell Memory1

PubMed Central

Dudani, Renu; Russell, Marsha; van Faassen, Henk; Krishnan, Lakshmi; Sad, Subash

2014-01-01

Fas death pathway is important for lymphocyte homeostasis, but the role of Fas pathway in T cell memory development is not clear. We show that whereas the expansion and contraction of CD8+ T cell response against Listeria monocytogenes were similar for wild-type (WT) and Fas ligand (FasL) mutant mice, the majority of memory CD8+ T cells in FasL mutant mice displayed an effector memory phenotype in the long-term in comparison with the mainly central memory phenotype displayed by memory CD8+ T cells in WT mice. Memory CD8+ T cells in FasL mutant mice expressed reduced levels of IFN-γ and displayed poor homeostatic and Ag-induced proliferation. Impairment in CD8+ T cell memory in FasL mutant hosts was not due to defective programming or the expression of mutant FasL on CD8+ T cells, but was caused by perturbed cytokine environment in FasL mutant mice. Although adoptively transferred WT memory CD8+ T cells mediated protection against L. monocytogenes in either the WT or FasL mutant hosts, FasL mutant memory CD8+ T cells failed to mediate protection even in WT hosts. Thus, in individuals with mutation in Fas pathway, impairment in the function of the memory CD8+ T cells may increase their susceptibility to recurrent/latent infections. PMID:18292515

Limonene encapsulation in freeze dried gellan systems.

PubMed

Evageliou, Vasiliki; Saliari, Dimitra

2017-05-15

The encapsulation of limonene in freeze-dried gellan systems was investigated. Surface and encapsulated limonene content was determined by measurement of the absorbance at 252nm. Gellan matrices were both gels and solutions. For a standard gellan concentration (0.5wt%) gelation was induced by potassium or calcium chloride. Furthermore, gellan solutions of varying concentrations (0.25-1wt%) were also studied. Limonene was added at two different concentrations (1 and 2mL/100g sample). Gellan gels encapsulated greater amounts of limonene than solutions. Among all gellan gels, the KCl gels had the greater encapsulated limonene content. However, when the concentration of limonene was doubled in these KCl gels, the encapsulated limonene decreased. The surface limonene content was significant, especially for gellan solutions. The experimental conditions and not the mechanical properties of the matrices were the dominant factor in the interpretation of the observed results. Copyright © 2016 Elsevier Ltd. All rights reserved.

Lead poisoning of passerines at a trap and skeet range

USGS Publications Warehouse

Vyas, N.B.; Spann, J.W.; Heinz, G.H.; Beyer, W.N.; Jaquette, J.A.; Mengelkoch, J.M.

2000-01-01

Our objective was to determine if ground foraging passerines in a woodland surrounding a trap and skeet range were subject to lead poisoning. Lead availability to birds was determined by shot counts and soil and earthworm analysis. Avian exposure to lead was identified by measuring free-erythrocyte protoporphyrin levels in blood and lead in tissues of three passerine species. Results showed that most shot were found in the top 3 cm of soil. Lead measurements ranged from 110 to 27,000 ppm (dry wt) in soil and were 660 and 840 ppm in earthworms. Sparrows held in an aviary at the range (p = 0.02) and free-flying juncos (p = 0.0005) mistnetted at the range displayed significantly higher protoporphyrin levels than those at an uncontaminated site. Sparrow and cowbird carcasses from the aviary carried 37 and 39 ppm lead (dry wt), respectively, whereas a junco liver contained 9.3 ppm lead.

Nanofibrillated cellulose (CNF) from eucalyptus sawdust as a dry strength agent of unrefined eucalyptus handsheets.

PubMed

Vallejos, María Evangelina; Felissia, Fernando Esteban; Area, María Cristina; Ehman, Nanci Vanesa; Tarrés, Quim; Mutjé, Pere

2016-03-30

Nanofibrillated cellulose has been obtained from the cellulosic fraction of eucalyptus sawdust. The fractionation process involved the partial removal of hemicelluloses and lignin. CNF was obtained using TEMPO oxidation with NaOCl in basic medium followed by mechanical homogenization. The obtained CNF was subsequently used as a dry strength agent on unbleached unrefined eucalyptus pulp. The addition of 3, 6 and 9 wt.% of CNF increased lineally the tensile index of handsheets to about 55 N mg(-1) at 35°SR, compatible with papermachine runnability. The other mechanical properties also increased substantially, and porosity decreased moderately. The estimated specific surface and average diameter of these CNF were 60 m(2)g(-1), and of 41.0 nm, respectively. The addition of 9 wt.% of CNF produced an increase in mechanical strength, equivalent to that produced by PFI refining at 1600 revolutions. Copyright © 2015 Elsevier Ltd. All rights reserved.

Poly(vinyl alcohol)-based film potentially suitable for antimicrobial packaging applications.

PubMed

Musetti, Alessandro; Paderni, Katia; Fabbri, Paola; Pulvirenti, Andrea; Al-Moghazy, Marwa; Fava, Patrizia

2014-04-01

This work aimed at developing a thin and water-resistant food-grade poly(vinyl alcohol) (PVOH)-based matrix able to swell when in contact with high moisture content food products without rupturing to release antimicrobial agents onto the food surface. This film was prepared by blending PVOH and 7.20% (wt/wt of PVOH) of poly(ethylene glycol) (PEG) with citric acid as crosslinking agent. The film-forming solution was then casted onto a flat surface and the obtained film was 60 μm in thickness and showed a good transparency (close to T = 100%) in the visible region (400 to 700 nm). After immersion in water for 72 h at room temperature, the crosslinked matrix loses only 19.2% of its original weight (the percentage includes the amount of unreacted crosslinking agent, antimicrobial in itself). Water content, degree of swelling, and crosslinking density of the film prove that the presence of PEG diminishes the hydrophilic behavior of the material. Also the mechanical properties of the wet and dry film were assessed. Alongside this, 2.5% (wt/wt of dry film) of grapefruit seed extract (GSE), an antimicrobial agent, was added to the film-forming solution just before casting and the ability of the plastic matrix to release the additive was then evaluated in vitro against 2 GSE-susceptible microorganisms, Salmonella enteritidis and Listeria innocua. The results indicate that the developed matrix may be a promising food-grade material for the incorporation of active substances. © 2014 Institute of Food Technologists®

A HER2-targeting antibody-drug conjugate, trastuzumab deruxtecan (DS-8201a), enhances antitumor immunity in a mouse model.

PubMed

Iwata, Tomomi Nakayama; Ishii, Chiaki; Ishida, Saori; Ogitani, Yusuke; Wada, Teiji; Agatsuma, Toshinori

2018-04-27

Trastuzumab deruxtecan (DS-8201a), a HER2-targeting antibody-drug conjugate with a topoisomerase I inhibitor exatecan derivative (DX-8951 derivative, DXd), has been reported to exert potent antitumor effects in xenograft mouse models and clinical trials. In this study, the immune system-activating ability of DS-8201a was assessed. DS-8201a significantly suppressed tumor growth in an immunocompetent mouse model with human HER2-expressing CT26.WT (CT26.WT-hHER2) cells. Cured immunocompetent mice rejected not only re-challenged CT26.WT-hHER2 cells, but also CT26.WT-mock cells. Splenocytes from the cured mice responded to both CT26.WT-hHER2 and CT26.WT-mock cells. Further analyses revealed that DXd up-regulated CD86 expression on bone marrow-derived DCs in vitro, and that DS-8201a increased tumor-infiltrating DCs and up-regulated their CD86 expression in vivo. DS-8201a also increased tumor-infiltrating CD8+ T cells and enhanced PD-L1 and MHC class I expression on tumor cells. Furthermore, combination therapy with DS-8201a and anti-PD-1 antibody was more effective than either monotherapy. In conclusion, DS-8201a enhanced antitumor immunity, as evidenced by the increased expression of DC markers, augmented expression of MHC class I in tumor cells, and rejection of re-challenged tumor cells by adaptive immune cells, suggesting that DS-8201a enhanced tumor recognition by T cells. Furthermore, DS-8201a treatment benefited from combination with anti-PD-1 antibody, possibly due to increased T cell activity and up-regulated PD-L1 expression induced by DS-8201a. Copyright ©2018, American Association for Cancer Research.

The contribution of Chlamydia-specific CD8⁺ T cells to upper genital tract pathology.

PubMed

Vlcek, Kelly R; Li, Weidang; Manam, Srikanth; Zanotti, Brian; Nicholson, Bruce J; Ramsey, Kyle H; Murthy, Ashlesh K

2016-02-01

Genital chlamydial infections lead to severe upper reproductive tract pathology in a subset of untreated women. We demonstrated previously that tumor necrosis factor (TNF)-α-producing CD8(+) T cells contribute significantly to chlamydial upper genital tract pathology in female mice. In addition, we observed that minimal chlamydial oviduct pathology develops in OT-1 transgenic (OT-1) mice, wherein the CD8(+) T-cell repertoire is restricted to recognition of the ovalbumin peptide Ova(257-264), suggesting that non-Chlamydia-specific CD8(+) T cells may not be responsible for chlamydial pathogenesis. In the current study, we evaluated whether antigen-specific CD8(+) T cells mediate chlamydial pathology. Groups of wild-type (WT) C57BL/6J, OT-1 mice, and OT-1 mice replete with WT CD8(+) T cells (1 × 10(6) cells per mouse intravenously) were infected intravaginally with C. muridarum (5 × 10(4) IFU/mouse). Serum total anti-Chlamydia antibody and total splenic anti-Chlamydia interferon (IFN)-γ and TNF-α responses were comparable among the three groups of animals. However, Chlamydia-specific IFN-γ and TNF-α production from purified splenic CD8(+) T cells of OT-1 mice was minimal, whereas responses in OT-1 mice replete with WT CD8(+) T cells were comparable to those in WT animals. Vaginal chlamydial clearance was comparable between the three groups of mice. Importantly, the incidence and severity of oviduct and uterine horn pathology was significantly reduced in OT-1 mice but reverted to WT levels in OT-1 mice replete with WT CD8(+) T cells. Collectively, these results demonstrate that Chlamydia-specific CD8(+) T cells contribute significantly to upper genital tract pathology.

Treatment with chemotherapy and dendritic cells pulsed with multiple Wilms' tumor 1 (WT1)-specific MHC class I/II-restricted epitopes for pancreatic cancer.

PubMed

Koido, Shigeo; Homma, Sadamu; Okamoto, Masato; Takakura, Kazuki; Mori, Masako; Yoshizaki, Shinji; Tsukinaga, Shintaro; Odahara, Shunichi; Koyama, Seita; Imazu, Hiroo; Uchiyama, Kan; Kajihara, Mikio; Arakawa, Hiroshi; Misawa, Takeyuki; Toyama, Yoichi; Yanagisawa, Satoru; Ikegami, Masahiro; Kan, Shin; Hayashi, Kazumi; Komita, Hideo; Kamata, Yuko; Ito, Masaki; Ishidao, Takefumi; Yusa, Sei-Ichi; Shimodaira, Shigetaka; Gong, Jianlin; Sugiyama, Haruo; Ohkusa, Toshifumi; Tajiri, Hisao

2014-08-15

We performed a phase I trial to investigate the safety, clinical responses, and Wilms' tumor 1 (WT1)-specific immune responses following treatment with dendritic cells (DC) pulsed with a mixture of three types of WT1 peptides, including both MHC class I and II-restricted epitopes, in combination with chemotherapy. Ten stage IV patients with pancreatic ductal adenocarcinoma (PDA) and 1 patient with intrahepatic cholangiocarcinoma (ICC) who were HLA-positive for A*02:01, A*02:06, A*24:02, DRB1*04:05, DRB1*08:03, DRB1*15:01, DRB1*15:02, DPB1*05:01, or DPB1*09:01 were enrolled. The patients received one course of gemcitabine followed by biweekly intradermal vaccinations with mature DCs pulsed with MHC class I (DC/WT1-I; 2 PDA and 1 ICC), II (DC/WT1-II; 1 PDA), or I/II-restricted WT1 peptides (DC/WT1-I/II; 7 PDA), and gemcitabine. The combination therapy was well tolerated. WT1-specific IFNγ-producing CD4(+) T cells were significantly increased following treatment with DC/WT1-I/II. WT1 peptide-specific delayed-type hypersensitivity (DTH) was detected in 4 of the 7 patients with PDA vaccinated with DC/WT1-I/II and in 0 of the 3 patients with PDA vaccinated with DC/WT1-I or DC/WT1-II. The WT1-specific DTH-positive patients showed significantly improved overall survival (OS) and progression-free survival (PFS) compared with the negative control patients. In particular, all 3 patients with PDA with strong DTH reactions had a median OS of 717 days. The activation of WT1-specific immune responses by DC/WT1-I/II combined with chemotherapy may be associated with disease stability in advanced pancreatic cancer. ©2014 American Association for Cancer Research.

Enantioselective oxidation of racemic lactic acid to D-lactic acid and pyruvic acid by Pseudomonas stutzeri SDM.

PubMed

Gao, Chao; Qiu, Jianhua; Li, Jingchen; Ma, Cuiqing; Tang, Hongzhi; Xu, Ping

2009-03-01

D-lactic acid and pyruvic acid are two important building block intermediates. Production of D-lactic acid and pyruvic acid from racemic lactic acid by biotransformation is economically interesting. Biocatalyst prepared from 9 g dry cell wt l(-1) of Pseudomonas stutzeri SDM could catalyze 45.00 g l(-1)DL-lactic acid into 25.23 g l(-1)D-lactic acid and 19.70 g l(-1) pyruvic acid in 10h. Using a simple ion exchange process, D-lactic acid and pyruvic acid were effectively separated from the biotransformation system. Co-production of d-lactic acid and pyruvic acid by enantioselective oxidation of racemic lactic acid is technically feasible.

Effects of chronic dietary lead in American kestrels (Falco sparverius)

USGS Publications Warehouse

Franson, J. Christian; Sileo, Louis; Pattee, Oliver H.; Moore, John F.

1983-01-01

American kestrels were fed a diet containing 0, 10, or 50 ppm lead (Pb) powder for at least 5 mo. Blood delta-aminolevulinic acid dehydratase (ALAD) activity in birds receiving 50 ppm Pb was as low as 20% of controls but no significant effects were noted in packed cell volume (PCV) or hemoglobin concentration (Hb). Mean liver Pb residues in birds fed 50 ppm Pb were 1.3 and 2.4 ppm (dry wt) for males and females, respectively. Liver Pb residues in birds fed 10 ppm Pb were not significantly greater than controls. There was no significant correlation between blood ALAD activity and blood Pb concentration, no consistent histopathological lesions were noted, and body and organ weights were not affected.

Tissue-specific root ion profiling reveals essential roles of the CAX and ACA calcium transport systems in response to hypoxia in Arabidopsis

PubMed Central

Wang, Feifei; Chen, Zhong-Hua; Liu, Xiaohui; Colmer, Timothy David; Zhou, Meixue; Shabala, Sergey

2016-01-01

Waterlogging is a major abiotic stress that limits the growth of plants. The crucial role of Ca2+ as a second messenger in response to abiotic and biotic stimuli has been widely recognized in plants. However, the physiological and molecular mechanisms of Ca2+ distribution within specific cell types in different root zones under hypoxia is poorly understood. In this work, whole-plant physiological and tissue-specific Ca2+ changes were studied using several ACA (Ca2+-ATPase) and CAX (Ca2+/proton exchanger) knock-out Arabidopsis mutants subjected to waterlogging treatment. In the wild-type (WT) plants, several days of hypoxia decreased the expression of ACA8, CAX4, and CAX11 by 33% and 50% compared with the control. The hypoxic treatment also resulted in an up to 11-fold tissue-dependent increase in Ca2+ accumulation in root tissues as revealed by confocal microscopy. The increase was much higher in stelar cells in the mature zone of Arabidopsis mutants with loss of function for ACA8, ACA11, CAX4, and CAX11. In addition, a significantly increased Ca2+ concentration was found in the cytosol of stelar cells in the mature zone after hypoxic treatment. Three weeks of waterlogging resulted in dramatic loss of shoot biomass in cax11 plants (67% loss in shoot dry weight), while in the WT and other transport mutants this decline was only 14–22%. These results were also consistent with a decline in leaf chlorophyll fluorescence (F v/F m). It is suggested that CAX11 plays a key role in maintaining cytosolic Ca2+ homeostasis and/or signalling in root cells under hypoxic conditions. PMID:26889007

Effects of the Loss of Conjunctival Muc16 on Corneal Epithelium and Stroma in Mice

PubMed Central

Shirai, Kumi; Okada, Yuka; Cheon, Dong-Joo; Miyajima, Masayasu; Behringer, Richard R.; Yamanaka, Osamu; Saika, Shizuya

2014-01-01

Purpose. To examine the role of conjunctival Muc16 in the homeostasis of the ocular surface epithelium and stroma using Muc16-null knockout (KO) mice. Methods. We used KO mice (n = 58) and C57/BL6 (WT) mice (n = 58). Histology and immunohistochemistry were employed to analyze the phenotypes in the ocular surface epithelium. The expression of phospho-Stat3, AP-1 components, interleukin 6 (IL-6), and tumor necrosis factor-α (TNFα) in the cornea and conjunctiva was examined. The shape of the nuclei of corneal epithelial cells was examined to evaluate intraepithelial cell differentiation. Epithelial cell proliferation was studied using bromo-deoxyuridine labeling. Finally, the wound healing of a round defect (2-mm diameter) in the corneal epithelium was measured. The keratocyte phenotype and macrophage invasion in the stroma were evaluated after epithelial repair. Results. The loss of Muc16 activated Stat3 signal, affected JunB signal, and upregulated the expression of IL-6 in the conjunctiva. Basal-like cells were observed in the suprabasal layer of the corneal epithelium with an increase in proliferation. The loss of Muc16 accelerated the wound healing of the corneal epithelium. The incidence of myofibroblast appearance and macrophage invasion were more marked in KO stroma than in WT stroma after epithelial repair. Conclusions. The loss of Muc16 in the conjunctiva affected the homeostasis of the corneal epithelium and stroma. The mechanism might include the upregulation of the inflammatory signaling cascade (i.e., Stat3 signal, and IL-6 expression in the KO conjunctiva). Current data provides insight into the research of the pathophysiology of dry eye syndrome. PMID:24812549

Quantitative Proteomics Analysis of the cAMP/Protein Kinase A Signaling Pathway

PubMed Central

2012-01-01

To define the proteins whose expression is regulated by cAMP and protein kinase A (PKA), we used a quantitative proteomics approach in studies of wild-type (WT) and kin- (PKA-null) S49 murine T lymphoma cells. We also compared the impact of endogenous increases in the level of cAMP [by forskolin (Fsk) and the phosphodiesterase inhibitor isobutylmethylxanthine (IBMX)] or by a cAMP analogue (8-CPT-cAMP). We identified 1056 proteins in WT and kin- S49 cells and found that 8-CPT-cAMP and Fsk with IBMX produced differences in protein expression. WT S49 cells had a correlation coefficient of 0.41 between DNA microarray data and the proteomics analysis in cells incubated with 8-CPT-cAMP for 24 h and a correlation coefficient of 0.42 between the DNA microarray data obtained at 6 h and the changes in protein expression after incubation with 8-CPT-cAMP for 24 h. Glutathione reductase (Gsr) had a higher level of basal expression in kin- S49 cells than in WT cells. Consistent with this finding, kin- cells are less sensitive to cell killing and generation of malondialdehyde than are WT cells incubated with H2O2. Cyclic AMP acting via PKA thus has a broad impact on protein expression in mammalian cells, including in the regulation of Gsr and oxidative stress. PMID:23110364

Progressive Oxidation of Pyrite in Five Bituminous Coal Samples: An As XANES and 57Fe Mossbauer Spectroscopic Study

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kolker,A.; Huggins, F.

2007-01-01

Naturally occurring pyrite commonly contains minor substituted metals and metalloids (As, Se, Hg, Cu, Ni, etc.) that can be released to the environment as a result of its weathering. Arsenic, often the most abundant minor constituent in pyrite, is a sensitive monitor of progressive pyrite oxidation in coal. To test the effect of pyrite composition and environmental parameters on the rate and extent of pyrite oxidation in coal, splits of five bituminous coal samples having differing amounts of pyrite and extents of As substitution in the pyrite, were exposed to a range of simulated weathering conditions over a period ofmore » 17 months. Samples investigated include a Springfield coal from Indiana (whole coal pyritic S = 2.13 wt.%; As in pyrite = detection limit (d.l.) to 0.06 wt.%), two Pittsburgh coal samples from West Virginia (pyritic S = 1.32-1.58 wt.%; As in pyrite = d.l. to 0.34 wt.%), and two samples from the Warrior Basin, Alabama (pyritic S = 0.26-0.27 wt.%; As in pyrite = d.l. to 2.72 wt.%). Samples were collected from active mine faces, and expected differences in the concentration of As in pyrite were confirmed by electron microprobe analysis. Experimental weathering conditions in test chambers were maintained as follows: (1) dry Ar atmosphere; (2) dry O{sub 2} atmosphere; (3) room atmosphere (relative humidity {approx}20-60%); and (4) room atmosphere with samples wetted periodically with double-distilled water. Sample splits were removed after one month, nine months, and 17 months to monitor the extent of As and Fe oxidation using As X-ray absorption near-edge structure (XANES) spectroscopy and {sup 57}Fe Mossbauer spectroscopy, respectively. Arsenic XANES spectroscopy shows progressive oxidation of pyritic As to arsenate, with wetted samples showing the most rapid oxidation. {sup 57}Fe Mossbauer spectroscopy also shows a much greater proportion of Fe{sup 3+} forms (jarosite, Fe{sup 3+} sulfate, FeOOH) for samples stored under wet conditions, but much less difference among samples stored under dry conditions in different atmospheres. The air-wet experiments show evidence of pyrite re-precipitation from soluble ferric sulfates, with As retention in the jarosite phase. Extents of As and Fe oxidation were similar for samples having differing As substitution in pyrite, suggesting that environmental conditions outweigh the composition and amount of pyrite as factors influencing the oxidation rate of Fe sulfides in coal.« less

IL-4 Is Required for Sex Differences in Vulnerability to Focal Ischemia in Mice.

PubMed

Xiong, Xiaoxing; Xu, Lijun; Wei, Liang; White, Robin E; Ouyang, Yi-Bing; Giffard, Rona G

2015-08-01

Interleukin (IL)-4 protects from middle cerebral artery occlusion in male mice. Females generally show less injury in response to the same ischemic challenge, but the underlying mechanisms are not fully understood. We tested the importance of IL-4 in female protection using IL-4 knockout (KO) mice. IL-4 KO and wild-type (WT) mice of both sexes were subjected to middle cerebral artery occlusion. Infarct volume was assessed by triphenyltetrazolium chloride staining and neurobehavioral outcome by neuroscore. T cell proliferation was assessed after Concanavalin A exposure. Ischemic brain immune cell populations were analyzed by fluorescence-activated cell sorting and immunostaining. Infarction in WT females during estrus and proestrus phases was significantly smaller than in males; neurological score was better. Infarction volume was larger and neurological score worse in IL-4 KO compared with WT in both sexes, with no sex difference. Proliferation of T cells was inhibited in WT females with higher proliferation and no sex difference in IL-4 KO. Macrophage numbers and total T cells in the ischemic hemisphere were lower in WT females, and monocytes increased markedly in IL-4 KOs with no sex difference. The reduced macrophage infiltration in WT-females was predominantly M2. Loss of IL-4 increased CD68+ and iNOS+ cells and reduced YM1+ and Arg1+ cells in both sexes. IL-4 is required for female neuroprotection during the estrus phase of the estrus cycle. Protected WT females show a predominance of M2-activated microglia/macrophages and reduced inflammatory infiltration. Increasing macrophage M2 polarization, with or without added inhibition of infiltration, may be a new approach to stroke treatment. © 2015 American Heart Association, Inc.

A cluster-randomized trial assessing the impact of school water, sanitation, and hygiene improvements on pupil enrollment and gender parity in enrollment.

PubMed

Garn, Joshua V; Greene, Leslie E; Dreibelbis, Robert; Saboori, Shadi; Rheingans, Richard D; Freeman, Matthew C

2013-10-01

We employed a cluster randomized trial design to measure the impact of a school based water, sanitation, and hygiene (WASH) improvement on pupil enrollment and on gender parity in enrollment, in primary schools in Nyanza Province, Kenya (2007-2009). Among schools with poor water access during the dry season, those that received a water supply, hygiene promotion and water treatment (HP&WT) and sanitation improvement, demonstrated increased enrollment (β=0.091 [0.009, 0.173] p=0.03), which translates to 26 additional pupils per school on average. The proportion of girls enrolled in school also increased by 4% (prevalence ratio (PR)=1.04 [1.00, 1.07] p=0.02). Among schools with better baseline water access during the dry season (schools that didn't receive a water source), we found no evidence of increased enrollment in schools that received a HP&WT intervention (β=0.016 [-0.039, 0.072] p=0.56) or the HP&WT and sanitation intervention (β=0.027 [-0.028, 0.082]p=0.34), and there was no evidence of improved gender parity (PR=0.99 [0.96, 1.02] p=0.59, PR=1.00 [0.97, 1.02] p=0.75, respectively). Our findings suggest that increased school enrollment and improved gender parity may be influenced by a comprehensive WASH program that includes an improved water source; schools with poor water access during the dry season may benefit most from these interventions.

Toxicity of Pb-contaminated soil to Japanese quail (Coturnix japonica) and the use of the blood-dietary Pb slope in risk assessment

USGS Publications Warehouse

Beyer, W. Nelson; Chen, Yu; Henry, Paula; May, Thomas; Mosby, David; Rattner, Barnett A.; Shearn-Bochsler, Valerie I.; Sprague, Daniel; Weber, John

2014-01-01

This study relates tissue concentrations and toxic effects of Pb in Japanese quail (Coturnix japonica) to the dietary exposure of soil-borne Pb associated with mining and smelting. From 0% to 12% contaminated soil, by weight, was added to 5 experimental diets (0.12 to 382 mg Pb/kg, dry wt) and fed to the quail for 6 weeks. Benchmark doses associated with a 50% reduction in delta-aminolevulinic acid dehydratase activity were 0.62 mg Pb/kg in the blood, dry wt, and 27 mg Pb/kg in the diet. Benchmark doses associated with a 20% increase in the concentration of erythrocyte protoporphyrin were 2.7 mg Pb/kg in the blood and 152 mg Pb/kg in the diet. The quail showed no other signs of toxicity (histopathological lesions, alterations in plasma–testosterone concentration, and body and organ weights). The relation of the blood Pb concentration to the soil Pb concentration was linear, with a slope of 0.013 mg Pb/kg of blood (dry wt) divided by mg Pb/kg of diet. We suggest that this slope is potentially useful in ecological risk assessments on birds in the same way that the intake slope factor is an important parameter in risk assessments of children exposed to Pb. The slope may also be used in a tissue-residue approach as an additional line of evidence in ecological risk assessment, supplementary to an estimate of hazard based on dietary toxicity reference values.

Biochemical composition of three species of unionid mussels after emersion

USGS Publications Warehouse

Greseth, Shari L.; Cope, W.G.; Rada, R.G.; Waller, D.L.; Bartsch, M.R.

2003-01-01

Freshwater mussels are emersed (exposed to air) during conservation activities such as surveys and relocations. Success of these activities depends upon the ability of mussels to survive emersion and to re-burrow in the substratum. We evaluated the acute sublethal effects of emersion on three species of unionid mussels [pocketbook, Lampsilis cardium (Rafinesque, 1820); pimpleback, Quadrula pustulosa pustulosa (I. Lea, 1831); spike, Elliptio dilatata (Rafinesque, 1820)] by measuring three biochemicals (carbohydrate, lipid, protein) indicative of biochemical function and energy storage. Mussels were acclimated in water at 25A?C and exposed to five air temperatures (15, 20, 25, 35 and 45A?C) for 15, 30 and 60 min. After emersion, mussels were returned to water at 25A?C and observed for 14 days. Samples of mantle tissue were taken after the 14-day postexposure period and analysed for carbohydrate, lipid and protein. Three-way analysis of variance (ANOVA) did not reveal consistent trends in carbohydrate, lipid or protein concentrations due to sex of mussels, duration of emersion, air temperature or their interaction terms that indicated biological compensation to stress. Overall mean carbohydrate concentrations were greatest (range 447a??615 mg/g dry wt) among the species, followed by protein (179a??289 mg/g dry wt) and lipids (26.7a??38.1 mg/g dry wt). These results have positive implications for conducting conservation activities, because emersion over the range of temperatures (15a??35A?C) and durations (15a??60 min) examined did not appear acutely harmful to mussels.

The effect of filler addition and oven temperature to the antioxidant quality in the drying of Physalis angulata leaf extract obtained by subcritical water extraction

NASA Astrophysics Data System (ADS)

Susanti, R. F.; Natalia, Desy

2016-11-01

In traditional medicine, Physalis angulata which is well known as ceplukan in Indonesia, has been utilized to cure several diseases by conventional extraction in hot water. The investigation of the Swietenia mahagoni extract activity in modern medicine typically utilized organic solvents such as ethanol, methanol, chloroform and hexane in extraction. In this research, subcritical water was used as a solvent instead of organic solvent to extract the Pysalis angulata leaf part. The focus of this research was the investigation of extract drying condition in the presence of filler to preserve the quality of antioxidant in Swietenia mahagoni extract. Filler, which is inert, was added to the extract during drying to help absorb the water while protect the extract from exposure in heat during drying. The effects of filler types, concentrations and oven drying temperatures were investigated to the antioxidant quality covering total phenol and antioxidant activity. Aerosil and microcrystalline cellulose (MCC) were utilized as fillers with concentration was varied from 0-30 wt% for MCC and 0-15 wt% for aerosil. The oven drying temperature was varied from 40-60 oC. The results showed that compare to extract dried without filler, total phenol and antioxidant activity were improved upon addition of filler. The higher the concentration of filler, the better the antioxidant; however it was limited by the homogeneity of filler in the extract. Both of the variables (oven temperature and concentration) played an important role in the improvement of extract quality of Swietenia mahagoni leaf. It was related to the drying time which can be minimized to protect the deterioration of extract from heat. In addition, filler help to provide the powder form of extract instead of the typical extract form which is sticky and oily.

Development of high refractive ZnS/PVP/PDMAA hydrogel nanocomposites for artificial cornea implants.

PubMed

Zhang, Quanyuan; Su, Kai; Chan-Park, Mary B; Wu, Hong; Wang, Dongan; Xu, Rong

2014-03-01

A series of high refractive index (RI) ZnS/PVP/PDMAA hydrogel nanocomposites containing ZnS nanoparticles (NPs) were successfully synthesized via a simple ultraviolet-light-initiated free radical co-polymerization method. The average diameter of the ZnS NPs is ∼ 3 nm and the NPs are well dispersed and stabilized in the PVP/PDMAA hydrogel matrix up to a high content of 60 wt.% in the hydrogel nanocomposites. The equilibrium water content of ZnS/PVP/PDMAA hydrogel nanocomposites varied from 82.0 to 66.8 wt.%, while the content of mercaptoethanol-capped ZnS NPs correspondingly varied from 30 to 60 wt.%. The resulting nanocomposites are clear and transparent and their RIs were measured to be as high as 1.58-1.70 and 1.38-1.46 in the dry and hydrated states, respectively, which can be tuned by varying the ZnS NPs content. In vitro cytotoxicity assays suggested that the introduction of ZnS NPs added little cytotoxicity to the PVP/PDMAA hydrogel and all the hydrogel nanocomposites exhibited minimal cytotoxicity towards common cells. The hydrogel nanocomposites implanted in rabbit eyes can be well tolerated over 3 weeks. Hence, the high RI ZnS/PVP/PDMAA hydrogel nanocomposites with adjustable RIs developed in this work might potentially be a candidate material for artificial corneal implants. Copyright © 2013 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

A 300,000-mol-wt intermediate filament-associated protein in baby hamster kidney (BHK-21) cells.

PubMed

Yang, H Y; Lieska, N; Goldman, A E; Goldman, R D

1985-02-01

Native intermediate filament (IF) preparations from the baby hamster kidney fibroblastic cell line (BHK-21) contain a number of minor polypeptides in addition to the IF structural subunit proteins desmin, a 54,000-mol-wt protein, and vimentin, a 55,000-mol-wt protein. A monoclonal antibody was produced that reached exclusively with a high molecular weight (300,000) protein representative of these minor proteins. Immunological methods and comparative peptide mapping techniques demonstrated that the 300,000-mol-wt species was biochemically distinct from the 54,000- and 55,000-mol-wt proteins. Double-label immunofluorescence observations on spread BHK cells using this monoclonal antibody and a rabbit polyclonal antibody directed against the 54,000- and 55,000-mol-wt proteins showed that the 300,000-mol-wt species co-distributed with IF in a fibrous pattern. In cells treated with colchicine or those in the early stages of spreading, double-labeling with these antibodies revealed the co-existence of the respective antigens in the juxtanuclear cap of IF that is characteristic of cells in these physiological states. After colchicine removal, or in the late stages of cell spreading, the 300,00-mol-wt species and the IF subunits redistributed to their normal, highly coincident cytoplasmic patterns. Ultrastructural localization by the immunogold technique using the monoclonal antibody supported the light microscopic findings in that the 300,000-mol-wt species was associated with IF in the several physiological and morphological cell states investigated. The gold particle pattern was less intimately associated with IF than that defined by anti-54/55 and was one of non-uniform distribution along IF, being clustered primarily at points of proximity between IF, where an amorphous, proteinaceous material was often the labeled element. Occasionally, "bridges" of label were seen extending outward from such clusters on IF. Gold particles were infrequently bound to microtubules, microfilaments, or other cellular organelles, and when so, IF were usually contiguous. During multiple cycles of in vitro disassembly/assembly of the IF from native preparations, the 300,000-mol-wt protein remained in the fraction containing the 54,000- and 55,000-mol-wt structural subunits, whether the latter were in the soluble state or pelleted as formed filaments. In keeping with the nomenclature developed for the microtubule-associated proteins (MAPs), the acronym IFAP-300K (intermediate filament associated protein) is proposed for this molecule.

Properties of wastepaper sludge in geopolymer mortars for masonry applications.

PubMed

Yan, Shiqin; Sagoe-Crentsil, Kwesi

2012-12-15

This paper presents the results of an investigation into the use of wastepaper sludge in geopolymer mortar systems for manufacturing construction products. The investigation was driven by the increasing demand for reuse options in paper-recycling industry. Both fresh and hardened geopolymer mortar properties are evaluated for samples incorporating dry wastepaper sludge, and the results indicate potential end-use benefits in building product manufacture. Addition of wastepaper sludge to geopolymer mortar reduces flow properties, primarily due to dry sludge absorbing water from the binder mix. The average 91-day compressive strength of mortar samples incorporating 2.5 wt% and 10 wt% wastepaper sludge respectively retained 92% and 52% of the reference mortar strength. However, contrary to the normal trend of increasing drying shrinkage with increasing paper sludge addition to Portland cement matrices, the corresponding geopolymer drying shrinkage decreased by 34% and 64%. Equally important, the water absorption of hardened geopolymer mortar decreased with increasing paper sludge content at ambient temperatures, providing good prospects of overall potential for wastepaper sludge incorporation in the production of building and masonry elements. The results indicate that, despite its high moisture absorbance due to the organic matter and residual cellulose fibre content, wastepaper sludge appears compatible with geopolymer chemistry, and hence serves as a potential supplementary additive to geopolymer cementitious masonry products. Copyright © 2012 Elsevier Ltd. All rights reserved.

The structure and productivity of the Thalassia testudinum community in Bon Accord Lagoon, Tobago.

PubMed

Juman, Rahanna A

2005-05-01

The Thalassia testudinum dominated seagrass community in the Buccoo Reef/ Bon Accord Lagoon Marine Park, measures 0.5 km(2) and is part of a contiguous coral reef, seagrass bed and mangrove swamp system in southwest Tobago. 7 testudinum coverage, productivity and percent turnover rates were measured from February 1998 to February 1999 at four sample locations, while total T. testudinum biomass was measured at two locations in the lagoon from 1992-2002. Productivity and turnover rates varied spatially and seasonally. They were higher in the back-reef area than in the mangrove-fringed lagoon, and were lowest at locations near to a sewage outfall. 7 testudinum coverage ranged from 6.6% in the lagoon to 68.5% in the back-reef area while productivity ranged from 3.9 to 4.9 g dry wt m(-2) d(-1) . Productivity and percentage turnover rates were higher in the dry season (January-June) than in the wet season (July-December). Productivity ranged from 3.0 in the wet season to 5.0 g dry wt m2 d-' in the dry season while percentage turnover rates ranged from 4.2% to 5.6%. Total Thalassia biomass and productivity in Bon Accord Lagoon were compared to six similar sites in the Caribbean that also participate in the Caribbean Coastal Marine Productivity Program (CARICOMP). This seagrass community is being negatively impacted by nutrient-enriched conditions.

B-1a transitional cells are phenotypically distinct and are lacking in mice deficient in IκBNS.

PubMed

Pedersen, Gabriel K; Àdori, Monika; Khoenkhoen, Sharesta; Dosenovic, Pia; Beutler, Bruce; Karlsson Hedestam, Gunilla B

2014-09-30

B-1 cells mediate early protection against infection by responding to T cell-independent (TI) antigens found on the surface of various pathogens. Mice with impaired expression of the atypical IκB protein IκBNS have markedly reduced frequencies of B-1 cells. We used a mouse strain with dysfunctional IκBNS derived from an N-ethyl-N-nitrosourea (ENU) screen, named bumble, to investigate the point in the development of B-1 cells where IκBNS is required. The presence of wild-type (wt) peritoneal cells in mixed wt/bumble chimeras did not rescue the development of bumble B-1 cells, but wt peritoneal cells transferred to bumble mice restored natural IgM levels and response to TI antigens. The bumble and wt mice displayed similar levels of fetal liver B-1 progenitors and splenic neonatal transitional B (TrB) cells, both of which were previously shown to give rise to B-1 cells. Interestingly, we found that a subset of wt neonatal TrB cells expressed common B-1a markers (TrB-1a) and that this cell population was absent in the bumble neonatal spleen. Sorted TrB-1a (CD93(+)IgM(+)CD5(+)) cells exclusively generated B-1a cells when adoptively transferred, whereas sorted CD93(+)IgM(+)CD5(-) cells gave rise to B-2 cells and, to a lesser extent, B-1b and B-1a cells. This study identifies a phenotypically distinct splenic population of TrB-1a cells and establishes that the development of B-1a cells is blocked before this stage in the absence of IκBNS.

Development of oral cancer vaccine using recombinant Bifidobacterium displaying Wilms' tumor 1 protein.

PubMed

Kitagawa, Koichi; Oda, Tsugumi; Saito, Hiroki; Araki, Ayame; Gonoi, Reina; Shigemura, Katsumi; Hashii, Yoshiko; Katayama, Takane; Fujisawa, Masato; Shirakawa, Toshiro

2017-06-01

Several types of vaccine-delivering tumor-associated antigens (TAAs) have been developed in basic and clinical research. Wilms' tumor 1 (WT1), identified as a gene responsible for pediatric renal neoplasm, is one of the most promising TAA for cancer immunotherapy. Peptide and dendritic cell-based WT1 cancer vaccines showed some therapeutic efficacy in clinical and pre-clinical studies but as yet no oral WT1 vaccine can be administrated in a simple and easy way. In the present study, we constructed a novel oral cancer vaccine using a recombinant Bifidobacterium longum displaying WT1 protein. B. longum 420 was orally administered into mice inoculated with WT1-expressing tumor cells for 4 weeks to examine anti-tumor effects. To analyze the WT1-specific cellular immune responses to oral B. longum 420, mice splenocytes were isolated and cytokine production and cytotoxic activities were determined. Oral administrations of B. longum 420 significantly inhibited WT1-expressing tumor growth and prolonged survival in mice. Immunohistochemical study and immunological assays revealed that B. longum 420 substantially induced tumor infiltration of CD4 + T and CD8 + T cells, systemic WT1-specific cytokine production, and cytotoxic activity mediated by WT1-epitope specific cytotoxic T lymphocytes, with no apparent adverse effects. Our novel oral cancer vaccine safely induced WT1-specific cellular immunity via activation of the gut mucosal immune system and achieved therapeutic efficacy with several practical advantages over existing non-oral vaccines.

The YAP1/SIX2 axis is required for DDX3-mediated tumor aggressiveness and cetuximab resistance in KRAS-wild-type colorectal cancer

PubMed Central

Wu, De-Wei; Lin, Po-Lin; Wang, Lee; Huang, Chi-Chou; Lee, Huei

2017-01-01

The mechanism underlying tumor aggressiveness and cetuximab (CTX) resistance in KRAS-wild-type (KRAS -WT) colorectal cancer remains obscure. We here provide evidence that DDX3 promoted soft agar growth and invasiveness of KRAS-WT cells, as already confirmed in KRAS-mutated cells. Mechanistically, increased KRAS expression induced ROS production, which elevated HIF-1α and YAP1 expression. Increased HIF-1α persistently promoted DDX3 expression via a KRAS/ROS/HIF-1α feedback loop. DDX3-mediated aggressiveness and CTX resistance were regulated by the YAP1/SIX2 axis in KRAS-WT cells and further confirmed in animal models. Kaplan-Meier and Cox regression analysis indicated that DDX3, KRAS, and YAP1 expression had prognostic value for OS and RFS in KRAS-WT and KRAS-mutated tumors, but SIX2 and YAP1/SIX2 were prognostic value only in KRAS-WT patients. The observation from patients seemed to support the mechanistic action of cell and animal models. We therefore suggest that combining YAP1 inhibitors with CTX may therefore suppress DDX3-mediated tumor aggressiveness and enhance CTX sensitivity in KRAS-WT colorectal cancer. PMID:28435452

A Novel Method for Assessing Sex-Specific and Genotype-Specific Response to Injury in Astrocyte Culture

PubMed Central

Liu, Mingyue; Oyarzabal, Esteban; Yang, Rui; Murphy, Stephanie J; Hurn, Patricia D.

2008-01-01

Female astrocytes sustain less cell death from oxygen-glucose deprivation (OGD) than male astrocytes. Arimidex, an aromatase inhibitor, abolishes these sex differences. To verify sex-dependent differences in P450 aromatase function in astrocyte cell death following OGD, we developed a novel method that uses sex-specific and genotype-specific single pup primary astrocyte cultures from wild-type (WT) and aromatase-knockout (ArKO) mice. After determining sex by external and internal examination as well as PCR and genotype by PCR amplification of tail cDNA, we established cultures from 1−3 day-old male and female, WT and ArKO mice pups and grew them to confluence in estrogen-free media. Cell death was measured by lactate dehydrogenase (LDH) assay. Our study shows that, while WT female astrocytes are more resistant to OGD than WT male cells, sex differences disappear in ArKO cells. Cell death is significantly increased in ArKO compared to WT in female astrocytes but not male cells. Therefore, P450 aromatase appears to be essential in endogenous neuroprotection in females, and this finding may have clinical implications. This innovative technique may also be applied to other in vitro studies of sex-related functional differences. PMID:18436308

Manipulation of culture strategies to enhance capsaicin biosynthesis in suspension and immobilized cell cultures of Capsicum chinense Jacq. cv. Naga King Chili.

PubMed

Kehie, Mechuselie; Kumaria, Suman; Tandon, Pramod

2014-06-01

Manipulation of culture strategies was adopted to study the influence of nutrient stress, pH stress and precursor feeding on the biosynthesis of capsaicin in suspension and immobilized cell cultures of C. chinense. Cells cultured in the absence of one of the four nutrients (ammonium and potassium nitrate for nitrate and potassium stress, potassium dihydrogen orthophosphate for phosphorus stress, and sucrose for sugar stress) influenced the accumulation of capsaicin. Among the stress factors studied, nitrate stress showed maximal capsaicin production on day 20 (505.9 ± 2.8 μg g(-1) f.wt) in immobilized cell, whereas in suspension cultures the maximum accumulation (345.5 ± 2.9 μg g(-1) f.wt) was obtained on day 10. Different pH affected capsaicin accumulation; enhanced accumulation of capsaicin (261.6 ± 3.4 μg g(-1) f.wt) was observed in suspension cultures at pH 6 on day 15, whereas in case of immobilized cultures the highest capsaicin content (433.3 ± 3.3 μg g(-1) f.wt) was obtained at pH 5 on day 10. Addition of capsaicin precursors and intermediates significantly enhanced the biosynthesis of capsaicin, incorporation of vanillin at 100 μM in both suspension and immobilized cell cultures resulted in maximum capsaicin content with 499.1 ± 5.5 μg g(-1) f.wt on day 20 and 1,315.3 ± 10 μg g(-1) f.wt on day 10, respectively. Among the different culture strategies adopted to enhance capsaicin biosynthesis in cell cultures of C. chinense, cells fed with vanillin resulted in the maximum capsaicin accumulation. The rate of capsaicin production was significantly higher in immobilized cells as compared to freely suspended cells.

Feasibility of hydrogel fiducial markers for in vivo proton range verification using PET

NASA Astrophysics Data System (ADS)

Cho, Jongmin; Campbell, Patrick; Wang, Min; Alqathami, Mamdooh; Mawlawi, Osama; Kerr, Matthew; Cho, Sang Hyun

2016-03-01

Biocompatible/biodegradable hydrogel polymers were immersed in 18O-enriched water and 16O-water to create 18O-water hydrogels and 16O-water hydrogels. In both cases, the hydrogels were made of ~91 wt% water and ~9 wt% polymer. In addition, 5-8 μm Zn powder was suspended in 16O-water and 18O-enriched water and cross-linked with hydrogel polymers to create Zn/16O-water hydrogels (30/70 wt%, ~9 wt% polymer) and Zn/18O-water hydrogels (10/90 wt%), respectively. A block of extra-firm ‘wet’ tofu (12.3  ×  8.8  ×  4.9 cm, ρ  ≈  1.05 g cm-3) immersed in water was injected with Zn/16O-water hydrogels (0.9 ml each) at four different depths using an 18-gauge needle. Similarly, Zn/18O-water hydrogels (0.9 ml) were injected into a second tofu phantom. As a reference, both 16O-water hydrogels (1.8 ml) and 18O-water hydrogels (0.9 ml) in Petri dishes were irradiated in a ‘dry’ environment. The hydrogels in the wet tofu phantoms and dry Petri dishes were scanned via CT and images were used for treatment planning. Then, they were positioned at the proton distal dose fall-off region and irradiated (2 Gy) followed by PET/CT imaging. Notably high PET signals were observed only in 18O-water hydrogels in the dry environment. The visibility of the Zn/16O-water hydrogels injected into the tofu phantom was outstanding in CT images, but these hydrogels provided no noticeable PET signals. The visibility of the Zn/18O-water hydrogels in the wet tofu were excellent on CT and moderate on PET; however, the PET signals were weaker than those in the dry environment, possibly owing to 18O-water leaching out. The hydrogel markers studied here could be used to develop universal PET/CT fiducial markers. Their PET visibility (attributed more to activated 18O-water than Zn) after proton irradiation can be used for proton therapy/range verification. More investigation is needed to slow down the leaching of 18O-water.

Desulfurization of Dibenzothiophene and Diesel Oils by a Newly Isolated Gordona Strain, CYKS1

PubMed Central

Rhee, Sung-Keun; Chang, Je Hwan; Chang, Yong Keun; Chang, Ho Nam

1998-01-01

A dibenzothiophene (DBT)-desulfurizing bacterial strain was isolated and identified as Gordona strain CYKS1. Strain CYKS1 was found to transform DBT to 2-hydroxybiphenyl via the 4S pathway and to be able to also use organic sulfur compounds other than DBT as a sole sulfur source. Its desulfurization activity was susceptible to sulfate repression. Active resting cells for desulfurization could be prepared only in the early growth phase. When two types of diesel oils, middle distillate unit feed (MDUF) and light gas oil (LGO) containing various organic sulfur compounds including DBT, were treated with resting cells of strain CYKS1 for 12 h, the total sulfur content significantly decreased, from 0.15% (wt/wt) to 0.06% (wt/wt) for MDUF and from 0.3% (wt/wt) to 0.25% (wt/wt) for LGO. The newly isolated strain CYKS1 is considered to have good potential for application in the biodesulfurization of fossil fuels. PMID:9603863

Biodiesel from dewatered wastewater sludge: a two-step process for a more advantageous production.

PubMed

Pastore, Carlo; Lopez, Antonio; Lotito, Vincenzo; Mascolo, Giuseppe

2013-07-01

Alternative approaches for obtaining biodiesel from municipal sludge have been successfully investigated. In order to avoid the expensive conventional preliminary step of sludge drying, dewatered sludge (TSS: 15wt%) was used as starting material. The best performance in terms of yield of fatty acid methyl esters (18wt%) with the lowest energy demand (17MJkgFAME(-1)) was obtained by a new two-step approach based on hexane extraction carried out directly on dewatered acidified (H2SO4) sludge followed by methanolysis of extracted lipids. It was found that sulphuric acid plays a key role in the whole process not only for the transesterification of glycerides but also for the production of new free fatty acids from soaps and their esterification with methanol. In addition to biodiesel production, the investigated process allows valorization of primary sludge as it turns it into a valuable source of chemicals, namely sterols (2.5wt%), aliphatic alcohols (0.8wt%) and waxes (2.3wt%). Copyright © 2013 Elsevier Ltd. All rights reserved.

Low VOC Barrier Coating for Industrial Maintenance

DTIC Science & Technology

2012-09-01

VOC Total Solids (wt) Total Solids (volume) Percent Pigment Stormer Viscosity Brookfield Viscosity Pot Life Sag Resistance Theoretical...Percent Pigment – Stormer Viscosity – Brookfield Viscosity – Pot Life – Sag Resistance – Theoretical Coverage – Drying Times – Mixing Ratio

SUMO-modified insulin-like growth factor 1 receptor (IGF-1R) increases cell cycle progression and cell proliferation.

PubMed

Lin, Yingbo; Liu, Hongyu; Waraky, Ahmed; Haglund, Felix; Agarwal, Prasoon; Jernberg-Wiklund, Helena; Warsito, Dudi; Larsson, Olle

2017-10-01

Increasing number of studies have shown nuclear localization of the insulin-like growth factor 1 receptor (nIGF-1R) in tumor cells and its links to adverse clinical outcome in various cancers. Any obvious cell physiological roles of nIGF-1R have, however, still not been disclosed. Previously, we reported that IGF-1R translocates to cell nucleus and modulates gene expression by binding to enhancers, provided that the receptor is SUMOylated. In this study, we constructed stable transfectants of wild type IGF1R (WT) and triple-SUMO-site-mutated IGF1R (TSM) using igf1r knockout mouse fibroblasts (R-). Cell clones (R-WT and R-TSM) expressing equal amounts of IGF-1R were selected for experiments. Phosphorylation of IGF-1R, Akt, and Erk upon IGF-1 stimulation was equal in R-WT and R-TSM. WT was confirmed to enter nuclei. TSM did also undergo nuclear translocation, although to a lesser extent. This may be explained by that TSM heterodimerizes with insulin receptor, which is known to translocate to cell nuclei. R-WT proliferated substantially faster than R-TSM, which did not differ significantly from the empty vector control. Upon IGF-1 stimulation G1-S-phase progression of R-WT increased from 12 to 38%, compared to 13 to 20% of R-TSM. The G1-S progression of R-WT correlated with increased expression of cyclin D1, A, and CDK2, as well as downregulation of p27. This suggests that SUMO-IGF-1R affects upstream mechanisms that control and coordinate expression of cell cycle regulators. Further studies to identify such SUMO-IGF-1R dependent mechanisms seem important. © 2017 The Authors. Journal of Cellular Physiology Published by Wiley Periodicals Inc.

The R882H DNMT3A Mutation Associated with AML Dominantly Inhibits WT DNMT3A by Blocking its Ability to Form Active Tetramers

PubMed Central

Russler-Germain, David A.; Spencer, David H.; Young, Margaret A.; Lamprecht, Tamara L.; Miller, Christopher A.; Fulton, Robert; Meyer, Matthew R.; Erdmann-Gilmore, Petra; Townsend, R. Reid; Wilson, Richard K.; Ley, Timothy J.

2014-01-01

Summary Somatic mutations in DNMT3A, which encodes a de novo DNA methyltransferase, are found in ~30% of normal karyotype acute myeloid leukemia (AML) cases. Most mutations are heterozygous and alter R882 within the catalytic domain (most commonly R882H), suggesting the possibility of dominant negative consequences. The methyltransferase activity of R882H DNMT3A is reduced by ~80% compared to the WT enzyme. In vitro mixing of WT and R882H DNMT3A does not affect the WT activity but co-expression of the two proteins in cells profoundly inhibits the WT enzyme by disrupting its ability to homotetramerize. AML cells with the R882H mutation have severely reduced de novo methyltransferase activity and focal hypomethylation at specific CpGs throughout AML cell genomes. PMID:24656771

Deletion of the COOH-Terminal Domain of CXC Chemokine Receptor 4 Leads to the Down-regulation of Cell-to-Cell Contact, Enhanced Motility and Proliferation in Breast Carcinoma Cells

PubMed Central

Ueda, Yukiko; Neel, Nicole F.; Schutyser, Evemie; Raman, Dayanidhi; Richmond, Ann

2009-01-01

The CXC chemokine receptor 4 (CXCR4) contributes to the metastasis of human breast cancer cells. The CXCR4 COOH-terminal domain (CTD) seems to play a major role in regulating receptor desensitization and down-regulation. We expressed either wild-type CXCR4 (CXCR4-WT) or CTD-truncated CXCR4 (CXCR4-ΔCTD) in MCF-7 human mammary carcinoma cells to determine whether the CTD is involved in CXCR4-modulated proliferation of mammary carcinoma cells. CXCR4-WT-transduced MCF-7 cells (MCF-7/CXCR4-WT cells) do not differ from vector-transduced MCF-7 control cells in morphology or growth rate. However, CXCR4-ΔCTD-transduced MCF-7 cells (MCF-7/CXCR4-ΔCTD cells) exhibit a higher growth rate and altered morphology, potentially indicating an epithelial-to-mesenchymal transition. Furthermore, extracellular signal-regulated kinase (ERK) activation and cell motility are increased in these cells. Ligand induces receptor association with β-arrestin for both CXCR4-WT and CXCR4-ΔCTD in these MCF-7 cells. Overexpressed CXCR4-WT localizes predominantly to the cell surface in unstimulated cells, whereas a significant portion of overexpressed CXCR4-ΔCTD resides intracellularly in recycling endosomes. Analysis with human oligomicroarray, Western blot, and immunohistochemistry showed that E-cadherin and Zonula occludens are down-regulated in MCF-7/CXCR4-ΔCTD cells. The array analysis also indicates that mesenchymal marker proteins and certain growth factor receptors are up-regulated in MCF-7/CXCR4-ΔCTD cells. These observations suggest that (a) the overexpression of CXCR4-ΔCTD leads to a gain-of-function of CXCR4-mediated signaling and (b) the CTD of CXCR4-WT may perform a feedback repressor function in this signaling pathway. These data will contribute to our understanding of how CXCR4-ΔCTD may promote progression of breast tumors to metastatic lesions. PMID:16740704

ROS Modifiers and NOX4 Affect the Expression of the Survivin-Associated Radio-Adaptive Response.

PubMed

Murley, Jeffrey S; Arbiser, Jack L; Weichselbaum, Ralph R; Grdina, David J

2018-04-13

The survivin-associated radio-adaptive response can be induced following exposure to ionizing radiation in the dose range from 5 to 100 mGy, and its magnitude of expression is dependent upon the TP53 mutational status of cells and ROS signaling. The purpose of the study was to investigate the potential role of ROS in the development of the survivin-associated adaptive response. Utilizing human colon carcinoma HCT116 TP53 wild type (WT) and HCT116 isogenic TP53 null mutant (Mut) cell cultures, the roles of inter- and intracellular ROS signaling on expression of the adaptive response as evidenced by changes in intracellular translocation of survivin measured by ELISA, and cell survival determined by a standard colony forming assay were investigated using ROS modifying agents that include emodin, N-acetyl-l-cysteine (NAC), fulvene-5, honokiol, metformin and rotenone. The role of NADPH oxidase 4 (NOX4) in the survivin-associated adaptive response was investigated by transfecting HCT116 cells, both WT and Mut, with two different NOX4 siRNA oligomers and Western blotting. A dose of 5 mGy or a 15min exposure to 50µM of the ROS producing drug emodin were equally effective in inducing a pro-survival adaptive response in TP53 WT and a radio-sensitization adaptive response in TP53 Mut HCT116 cells. Each response was associated with a corresponding translocation of survivin into the cytoplasm or nucleus, respectively. Exposure to 10mM NAC completely inhibited both responses. Exposure to 10µM honokiol induced responses similar to those observed following NAC exposure in TP53 WT and Mut cells. The mitochondrial complex 1 inhibitor rotenone was effective in reducing both cytoplasmic and nuclear survivin levels, but was ineffective in altering the expression of the adaptive response in either TP53 WT or Mut cells. In contrast, both metformin and fulvene-5, inhibitors of NOX4, facilitated the reversal of TP53 WT and Mut adaptive responses from pro-survival to radio-sensitization and vice versa, respectively. These changes were accompanied by corresponding reversals in the translocation of survivin to the nuclei of TP53 WT and to the cytoplasm of TP53 Mut cells. The potential role of NOX4 in the expression of the survivin-associated adaptive response was investigated by transfecting HCT116 cells with NOX4 siRNA oligomers to inhibit NOX4 expression. Under these conditions NOX4 expression was inhibited by about 50%, resulting in a reversal in the expression of the TP53 WT and Mut survivin-associated adaptive responses as was observed following metformin and fulvene-5 treatment. Exposure to 5 mGy resulted in enhanced NOX4 expression by about 40% in both TP53 WT and Mut cells, in contrast to only a 1 to 2% increase following a 2Gy only exposure. Utilizing mixed cultures of HCT116 TP53 WT and isogenic null Mut cells, as few as 10% TP53 Mut cells were sufficient to control the expression of the remaining 90% WT cells and resulted in an overall radio-sensitization response accompanied by the nuclear translocation of survivin characteristic of homogeneous TP53 Mut populations. Copyright © 2018. Published by Elsevier Inc.

Induction of the Hajdu-Cheney Syndrome Mutation in CD19 B Cells in Mice Alters B-Cell Allocation but Not Skeletal Homeostasis.

PubMed

Yu, Jungeun; Zanotti, Stefano; Schilling, Lauren; Schoenherr, Chris; Economides, Aris N; Sanjay, Archana; Canalis, Ernesto

2018-06-01

Mice harboring Notch2 mutations replicating Hajdu-Cheney syndrome (Notch2 tm1.1ECan ) have osteopenia and exhibit an increase in splenic marginal zone B cells with a decrease in follicular B cells. Whether the altered B-cell allocation is responsible for the osteopenia of Notch2 tm1.1ECan mutants is unknown. To determine the effect of NOTCH2 activation in B cells on splenic B-cell allocation and skeletal phenotype, a conditional-by-inversion (COIN) Hajdu-Cheney syndrome allele of Notch2 (Notch2 [ΔPEST]COIN ) was used. Cre recombination generates a permanent Notch2 ΔPEST allele expressing a transcript for which sequences coding for the proline, glutamic acid, serine, and threonine-rich (PEST) domain are replaced by a stop codon. CD19-Cre drivers were backcrossed into Notch2 [ΔPEST]COIN/[ΔPEST]COIN to generate CD19-specific Notch2 ΔPEST/ΔPEST mutants and control Notch2 [ΔPEST]COIN/[ΔPEST]COIN littermates. There was an increase in marginal zone B cells and a decrease in follicular B cells in the spleen of CD19 Cre/WT ;Notch2 ΔPEST/ΔPEST mice, recapitulating the splenic phenotype of Notch2 tm1.1ECan mice. The effect was reproduced when the NOTCH1 intracellular domain was induced in CD19-expressing cells (CD19 Cre/WT ;Rosa Notch1/WT mice). However, neither CD19 Cre/WT ;Notch2 ΔPEST/ΔPEST nor CD19 Cre/WT ;Rosa Notch1/WT mice had a skeletal phenotype. Moreover, splenectomies in Notch2 tm1.1ECan mice did not reverse their osteopenic phenotype. In conclusion, Notch2 activation in CD19-expressing cells determines B-cell allocation in the spleen but has no skeletal consequences. Copyright © 2018 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.

Epimorphin Regulates the Intestinal Stem Cell Niche via Effects on the Stromal Microenvironment.

PubMed

Vishy, Courtney E; Swietlicki, Elzbieta A; Gazit, Vered; Amara, Suneetha; Heslop, Gabriela; Lu, Jianyun; Levin, Marc S; Rubin, Deborah C

2018-04-06

Stem cell therapy is a potential therapeutic approach for disorders characterized by intestinal injury or loss of functional surface area. Stem cell function and proliferation are mediated by the stem cell niche. Stromal cells such as intestinal subepithelial myofibroblasts (ISEMFs) are important but poorly studied components of the stem cell niche. To examine the role of ISEMFs, we have previously generated mice with deletion of epimorphin (Epim), an ISEMF protein and member of the syntaxin family of intracellular vesicle docking proteins that regulate cell secretion. Herein we explore the mechanisms for previous observations that Epim deletion increases gut crypt cell proliferation, crypt fission and small bowel length in vivo. Stem cell derived crypt culture techniques were used to explore the interaction between enteroids and myofibroblasts from Epim -/- and WT mice. Enteroids co-cultured with ISEMFS had increased growth and crypt-like budding compared to enteroids cultured without stromal support. Epim deletion in ISEMFs resulted in increased enteroid budding and surface area compared to co-cultures with WT ISEMFs. In primary crypt cultures, Epim -/- enteroids had significantly increased surface area and budding compared WTs. However stem cell assays comparing the number of Epim -/- vs WT colony forming units after first passage showed no differences in the absence of ISEMF support. Epim -/- vs. WT ISEMFs had increased Wnt4 expression and addition of Wnt4 to WT co-cultures enhanced budding. We conclude that ISEMFs play an important role in the stem cell niche. Epim regulates stem cell proliferation and differentiation via stromal contributions to the niche microenvironment.

A case of mistaken identity: CD11c-eYFP(+) cells in the normal mouse brain parenchyma and neural retina display the phenotype of microglia, not dendritic cells.

PubMed

Dando, Samantha J; Naranjo Golborne, Cecilia; Chinnery, Holly R; Ruitenberg, Marc J; McMenamin, Paul G

2016-08-01

Under steady-state conditions the central nervous system (CNS) is traditionally thought to be devoid of antigen presenting cells; however, putative dendritic cells (DCs) expressing enhanced yellow fluorescent protein (eYFP) are present in the retina and brain parenchyma of CD11c-eYFP mice. We previously showed that these mice carry the Crb1(rd8) mutation, which causes retinal dystrophic lesions; therefore we hypothesized that the presence of CD11c-eYFP(+) cells within the CNS may be due to pathology associated with the Crb1(rd8) mutation. We generated CD11c-eYFP Crb1(wt/wt) mice and compared the distribution and immunophenotype of CD11c-eYFP(+) cells in CD11c-eYFP mice with and without the Crb1(rd8) mutation. The number and distribution of CD11c-eYFP(+) cells in the CNS was similar between CD11c-eYFP Crb1(wt/wt) and CD11c-eYFP Crb1(rd8/rd8) mice. CD11c-eYFP(+) cells were distributed throughout the inner retina, and clustered in brain regions that receive input from the external environment or lack a blood-brain barrier. CD11c-eYFP(+) cells within the retina and cerebral cortex of CD11c-eYFP Crb1(wt/wt) mice expressed CD11b, F4/80, CD115 and Iba-1, but not DC or antigen presentation markers, whereas CD11c-eYFP(+) cells within the choroid plexus and pia mater expressed CD11c, I-A/I-E, CD80, CD86, CD103, DEC205, CD8α and CD135. The immunophenotype of CD11c-eYFP(+) cells and microglia within the CNS was similar between CD11c-eYFP Crb1(wt/wt) and CD11c-eYFP Crb1(rd8/rd8) mice; however, CD11c and I-A/I-E expression was significantly increased in CD11c-eYFP Crb1(rd8/rd8) mice. This study demonstrates that the overwhelming majority of CNS CD11c-eYFP(+) cells do not display the phenotype of DCs or their precursors and are most likely a subpopulation of microglia. GLIA 2016. GLIA 2016;64:1331-1349. © 2016 Wiley Periodicals, Inc.

Wilms tumor gene product: sensitive and contextually specific marker of serous carcinomas of ovarian surface epithelial origin.

PubMed

Hwang, Harry; Quenneville, Louise; Yaziji, Hadi; Gown, Allen M

2004-06-01

Carcinomas of ovarian surface epithelial origin can arise from, and often present at, extraovarian sites. There are few available markers for the positive identification of carcinomas of ovarian surface epithelial origin, which might aid in distinguishing them from metastatic carcinomas, such as of breast, colon, or lung origin. Recently, the Wilms tumor gene product (WT-1) has been shown to be expressed in ovarian surface and mesothelial epithelium. We tested the hypothesis that WT-1 would be a sensitive and specific marker of ovarian surface epithelium carcinomas. An archived series of 116 ovarian carcinomas (57 serous [43 ovarian, 14 extraovarian], 31 mucinous, 15 clear cell, 13 endometrioid), 118 breast carcinomas, 46 colonic carcinomas, and 45 nonsmall cell lung cancers were selected. A polyclonal antibody to the WT-1 gene product was applied to deparaffinized, formalin-fixed tissue sections after epitope retrieval. Fifty-two of 57 (93%) serous carcinomas of ovarian surface epithelial origin were WT-1-positive, in a nuclear pattern, with virtually all the tumor cell population positive in the majority of cases. None of the mucinous, clear cell, or endometrioid ovarian cancers were positive, and only 8 of 118 breast, 0 of 46 colonic, and 0 of 45 lung nonsmall cell carcinomas were WT-1-positive. These findings demonstrate that WT-1 is a highly sensitive and specific marker of serous carcinomas of ovarian surface epithelial origin (both ovarian and extraovarian). These results also contradict recent reports demonstrating WT-1 expression in both breast and lung carcinomas.

Dynamin-mediated Nephrin Phosphorylation Regulates Glucose-stimulated Insulin Release in Pancreatic Beta Cells*

PubMed Central

Jeon, Jongmin; Leibiger, Ingo; Moede, Tilo; Walter, Britta; Faul, Christian; Maiguel, Dony; Villarreal, Rodrigo; Guzman, Johanna; Berggren, Per-Olof; Mundel, Peter; Ricordi, Camillo; Merscher-Gomez, Sandra; Fornoni, Alessia

2012-01-01

We have previously demonstrated a role for Nephrin in glucose stimulated insulin release (GSIR). We now hypothesize that Nephrin phosphorylation is required for GSIR and that Dynamin influences Nephrin phosphorylation and function. MIN6-C3 Nephrin-deficient pancreatic beta cells and human islets were transfected with WT-Nephrin or with a mutant Nephrin in which the tyrosine residues responsible for SH2 domain binding were substituted with phenylalanine (3YF-Nephrin). GSIR and live images of Nephrin and vesicle trafficking were studied. Immunoprecipitation experiments and overexpression of WT-Dynamin or dominant negative Dynamin mutant (K44A-Dynamin) in WT-Nephrin, 3YF-Nephrin, or Nephrin siRNA-transfected cells were utilized to study Nephrin-Dynamin interaction. In contrast to WT-Nephrin or to single tyrosine mutants, 3YF-Nephrin did not positively affect GSIR and led to impaired cell-cell contacts and vesicle trafficking. K44A-Dynamin prevented the effect of Nephrin on GSIR in the absence of protein-protein interaction between Nephrin and Dynamin. Nephrin gene silencing abolished the positive effects of WT-Dynamin on GSIR. The effects of protamine sulfate and vanadate on Nephrin phosphorylation and GSIR were studied in MIN6 cells and human islets. WT-Nephrin phosphorylation after glucose occurred at Tyr-1176/1193 and resulted in improved GSIR. On the contrary, protamine sulfate-induced phosphorylation at Tyr-1176/1193/1217 was associated with Nephrin degradation and impaired GSIR. Vanadate, which prevented Nephrin dephosphorylation after glucose stimulation, improved GSIR in human islets and MIN6 cells. In conclusion, Dynamin-dependent Nephrin phosphorylation occurs in response to glucose and is necessary for Nephrin-mediated augmentation of GSIR. Pharmacological modulation of Nephrin phosphorylation may thus facilitate pancreatic beta cell function. PMID:22718751

Microscopic and Spectroscopic Analyses of Chlorhexidine Tolerance in Delftia acidovorans Biofilms

PubMed Central

Rema, Tara; Lawrence, John R.; Dynes, James J.; Hitchcock, Adam P.

2014-01-01

The physicochemical responses of Delftia acidovorans biofilms exposed to the commonly used antimicrobial chlorhexidine (CHX) were examined in this study. A CHX-sensitive mutant (MIC, 1.0 μg ml−1) was derived from a CHX-tolerant (MIC, 15.0 μg ml−1) D. acidovorans parent strain using transposon mutagenesis. D. acidovorans mutant (MT51) and wild-type (WT15) strain biofilms were cultivated in flow cells and then treated with CHX at sub-MIC and inhibitory concentrations and examined by confocal laser scanning microscopy (CLSM), scanning transmission X-ray microscopy (STXM), and infrared (IR) spectroscopy. Specific morphological, structural, and chemical compositional differences between the CHX-treated and -untreated biofilms of both strains were observed. Apart from architectural differences, CLSM revealed a negative effect of CHX on biofilm thickness in the CHX-sensitive MT51 biofilms relative to those of the WT15 strain. STXM analyses showed that the WT15 biofilms contained two morphochemical cell variants, whereas only one type was detected in the MT51 biofilms. The cells in the MT51 biofilms bioaccumulated CHX to a similar extent as one of the cell types found in the WT15 biofilms, whereas the other cell type in the WT15 biofilms did not bioaccumulate CHX. STXM and IR spectral analyses revealed that CHX-sensitive MT51 cells accumulated the highest levels of CHX. Pretreating biofilms with EDTA promoted the accumulation of CHX in all cells. Thus, it is suggested that a subpopulation of cells that do not accumulate CHX appear to be responsible for greater CHX resistance in D. acidovorans WT15 biofilm in conjunction with the possible involvement of bacterial membrane stability. PMID:25022584

Effect of Nano-Si3N4 Additives and Plasma Treatment on the Dry Sliding Wear Behavior of Plasma Sprayed Al2O3-8YSZ Ceramic Coatings

NASA Astrophysics Data System (ADS)

Gou, Junfeng; Zhang, Jian; Zhang, Qiwen; Wang, You; Wang, Chaohui

2017-04-01

In this paper, the effect of nano-Si3N4 additives and plasma treatment on the wear behavior of Al2O3-8YSZ ceramic coatings was studied. Nano-Al2O3, nano-8YSZ (8 wt.% Y2O3-stabilized ZrO2) and nano-Si3N4 powders were used as raw materials to fabricate four types of sprayable feedstocks. Plasma treatment was used to improve the properties of the feedstocks. The surface morphologies of the ceramic coatings were observed. The mechanical properties of the ceramic coatings were measured. The dry sliding wear behavior of the Al2O3-8YSZ coatings with and without Si3N4 additives was studied. Nano-Si3N4 additives and plasma treatment can improve the morphologies of the coatings by prohibiting the initiation of micro-cracks and reducing the unmelted particles. The hardness and bonding strength of AZSP (Al2O3-18 wt.% 8YSZ-10 wt.% Si3N4-plasma treatment) coating increased by 79.2 and 44% compared to those of AZ (Al2O3-20 wt.% 8YSZ) coating. The porosity of AZSP coating decreased by 85.4% compared to that of AZ coating. The wear test results showed that the addition of nano-Si3N4 and plasma treatment could improve the wear resistance of Al2O3-8YSZ coatings.

Sintered bentonite ceramics for the immobilization of cesium- and strontium-bearing radioactive waste

NASA Astrophysics Data System (ADS)

Ortega, Luis Humberto

The Advanced Fuel Cycle Initiative (AFCI) is a Department of Energy (DOE) program, that has been investigating technologies to improve fuel cycle sustainability and proliferation resistance. One of the program's goals is to reduce the amount of radioactive waste requiring repository disposal. Cesium and strontium are two primary heat sources during the first 300 years of spent nuclear fuel's decay, specifically isotopes Cs-137 and Sr-90. Removal of these isotopes from spent nuclear fuel will reduce the activity of the bulk spent fuel, reducing the heat given off by the waste. Once the cesium and strontium are separated from the bulk of the spent nuclear fuel, the isotopes must be immobilized. This study is focused on a method to immobilize a cesium- and strontium-bearing radioactive liquid waste stream. While there are various schemes to remove these isotopes from spent fuel, this study has focused on a nitric acid based liquid waste. The waste liquid was mixed with the bentonite, dried then sintered. To be effective sintering temperatures from 1100 to 1200°C were required, and waste concentrations must be at least 25 wt%. The product is a leach resistant ceramic solid with the waste elements embedded within alumino-silicates and a silicon rich phase. The cesium is primarily incorporated into pollucite and the strontium into a monoclinic feldspar. The simulated waste was prepared from nitrate salts of stable ions. These ions were limited to cesium, strontium, barium and rubidium. Barium and rubidium will be co-extracted during separation due to similar chemical properties to cesium and strontium. The waste liquid was added to the bentonite clay incrementally with drying steps between each addition. The dry powder was pressed and then sintered at various temperatures. The maximum loading tested is 32 wt. percent waste, which refers to 13.9 wt. percent cesium, 12.2 wt. percent barium, 4.1 wt. percent strontium, and 2.0 wt. percent rubidium. Lower loadings of waste were also tested. The final solid product was a hard dense ceramic with a density that varied from 2.12 g/cm3 for a 19% waste loading with a 1200°C sintering temperature to 3.03 g/cm 3 with a 29% waste loading and sintered at 1100°C. Differential Scanning Calorimetry and Thermal Gravimetric Analysis (DSC-TGA) of the loaded bentonite displayed mass loss steps which were consistent with water losses in pure bentonite. Water losses were complete after dehydroxylation at ˜650°C. No mass losses were evident beyond the dehydroxylation. The ceramic melts at temperatures greater than 1300°C. Light flash analysis found heat capacities of the ceramic to be comparable to those of strontium and barium feldspars as well as pollucite. Thermal conductivity improved with higher sintering temperatures, attributed to lower porosity. Porosity was minimized in 1200°C sinterings. Ceramics with waste loadings less than 25 wt% displayed slump, the lowest waste loading, 15 wt% bloated at a 1200°C sintering. Waste loading above 25 wt% produced smooth uniform ceramics when sintered >1100°C. Sintered bentonite may provide a simple alternative to vitrification and other engineered radioactive waste-forms.

Establishment of a Conditionally Immortalized Wilms Tumor Cell Line with a Homozygous WT1 Deletion within a Heterozygous 11p13 Deletion and UPD Limited to 11p15.

PubMed

Brandt, Artur; Löhers, Katharina; Beier, Manfred; Leube, Barbara; de Torres, Carmen; Mora, Jaume; Arora, Parineeta; Jat, Parmjit S; Royer-Pokora, Brigitte

2016-01-01

We describe a stromal predominant Wilms tumor with focal anaplasia and a complex, tumor specific chromosome 11 aberration: a homozygous deletion of the entire WT1 gene within a heterozygous 11p13 deletion and an additional region of uniparental disomy (UPD) limited to 11p15.5-p15.2 including the IGF2 gene. The tumor carried a heterozygous p.T41A mutation in CTNNB1. Cells established from the tumor carried the same chromosome 11 aberration, but a different, homozygous p.S45Δ CTNNB1 mutation. Uniparental disomy (UPD) 3p21.3pter lead to the homozygous CTNNB1 mutation. The tumor cell line was immortalized using the catalytic subunit of human telomerase (hTERT) in conjunction with a novel thermolabile mutant (U19dl89-97tsA58) of SV40 large T antigen (LT). This cell line is cytogenetically stable and can be grown indefinitely representing a valuable tool to study the effect of a complete lack of WT1 in tumor cells. The origin/fate of Wilms tumors with WT1 mutations is currently poorly defined. Here we studied the expression of several genes expressed in early kidney development, e.g. FOXD1, PAX3, SIX1, OSR1, OSR2 and MEIS1 and show that these are expressed at similar levels in the parental and the immortalized Wilms10 cells. In addition the limited potential for muscle/ osteogenic/ adipogenic differentiation similar to all other WT1 mutant cell lines is also observed in the Wilms10 tumor cell line and this is retained in the immortalized cells. In summary these Wilms10 cells are a valuable model system for functional studies of WT1 mutant cells.

Establishment of a Conditionally Immortalized Wilms Tumor Cell Line with a Homozygous WT1 Deletion within a Heterozygous 11p13 Deletion and UPD Limited to 11p15

PubMed Central

Brandt, Artur; Löhers, Katharina; Beier, Manfred; Leube, Barbara; de Torres, Carmen; Mora, Jaume; Arora, Parineeta; Jat, Parmjit S.; Royer-Pokora, Brigitte

2016-01-01

We describe a stromal predominant Wilms tumor with focal anaplasia and a complex, tumor specific chromosome 11 aberration: a homozygous deletion of the entire WT1 gene within a heterozygous 11p13 deletion and an additional region of uniparental disomy (UPD) limited to 11p15.5-p15.2 including the IGF2 gene. The tumor carried a heterozygous p.T41A mutation in CTNNB1. Cells established from the tumor carried the same chromosome 11 aberration, but a different, homozygous p.S45Δ CTNNB1 mutation. Uniparental disomy (UPD) 3p21.3pter lead to the homozygous CTNNB1 mutation. The tumor cell line was immortalized using the catalytic subunit of human telomerase (hTERT) in conjunction with a novel thermolabile mutant (U19dl89-97tsA58) of SV40 large T antigen (LT). This cell line is cytogenetically stable and can be grown indefinitely representing a valuable tool to study the effect of a complete lack of WT1 in tumor cells. The origin/fate of Wilms tumors with WT1 mutations is currently poorly defined. Here we studied the expression of several genes expressed in early kidney development, e.g. FOXD1, PAX3, SIX1, OSR1, OSR2 and MEIS1 and show that these are expressed at similar levels in the parental and the immortalized Wilms10 cells. In addition the limited potential for muscle/ osteogenic/ adipogenic differentiation similar to all other WT1 mutant cell lines is also observed in the Wilms10 tumor cell line and this is retained in the immortalized cells. In summary these Wilms10 cells are a valuable model system for functional studies of WT1 mutant cells. PMID:27213811

Electrode for molten carbonate fuel cell

DOEpatents

Iacovangelo, Charles D.; Zarnoch, Kenneth P.

1983-01-01

A sintered porous electrode useful for a molten carbonate fuel cell is produced which is composed of a plurality of 5 wt. % to 95 wt. % nickel balance copper alloy encapsulated ceramic particles sintered together by the alloy.

Synthetic Lethality of a Novel Small Molecule Against Mutant KRAS-Expressing Cancer Cells Involves AKT-Dependent ROS Production.

PubMed

Iskandar, Kartini; Rezlan, Majidah; Yadav, Sanjiv Kumar; Foo, Chuan Han Jonathan; Sethi, Gautam; Qiang, Yu; Bellot, Gregory L; Pervaiz, Shazib

2016-05-10

We recently reported the death-inducing activity of a small-molecule compound, C1, which triggered reactive oxygen species (ROS)-dependent autophagy-associated apoptosis in a variety of human cancer cell lines. In this study, we examine the ability of the compound to specifically target cancer cells harboring mutant KRAS with minimal activity against wild-type (WT) RAS-expressing cells. HCT116 cells expressing mutated KRAS are susceptible, while the WT-expressing HT29 cells are resistant. Interestingly, C1 triggers activation of mutant RAS, which results in the downstream phosphorylation and activation of AKT/PKB. Gene knockdown of KRAS or AKT or their pharmacological inhibition resulted in the abrogation of C1-induced ROS production and rescued tumor colony-forming ability. We also made use of HCT116 mutant KRAS knockout (KO) cells, which express only a single WT KRAS allele. Exposure of KO cells to C1 failed to increase mitochondrial ROS and cell death, unlike the parental cells harboring mutant KRAS. Similarly, mutant KRAS-transformed prostate epithelial cells (RWPE-1-RAS) were more sensitive to the ROS-producing and death-inducing effects of C1 than the vector only expressing RWPE-1 cells. An in vivo model of xenograft tumors generated with HCT116 KRAS(WT/MUT) or KRAS(WT/-) cells showed the efficacy of C1 treatment and its ability to affect the relative mitotic index in tumors harboring KRAS mutant. These data indicate a synthetic lethal effect against cells carrying mutant KRAS, which could have therapeutic implications given the paucity of KRAS-specific chemotherapeutic strategies. Antioxid. Redox Signal. 24, 781-794.

The Wilms tumor protein WT1 stimulates transcription of the gene encoding insulin-like growth factor binding protein 5 (IGFBP5).

PubMed

Müller, Miriam; Persson, Anja Bondke; Krueger, Katharina; Kirschner, Karin M; Scholz, Holger

2017-07-01

Insulin-like growth factor (IGF) binding proteins (IGFBPs) constitute a family of six secreted proteins that regulate the signaling of insulin-like growth factors (IGFs). IGFBP5 is the most conserved family member in vertebrates and the major IGF binding protein in bone. IGFBP5 is required for normal development of the musculoskeletal system, and various types of cancer frequently express high levels of IGFP5. Here we identify the gene encoding IGFBP5 as a novel downstream target of the Wilms tumor protein WT1. IGFBP5 and WT1 are expressed in an overlapping pattern in the condensing metanephric mesenchyme of embryonic murine kidneys. Down-regulation of WT1 by transfection with antisense vivo-morpholino significantly decreased Igfbp5 transcripts in murine embryonic kidney explants. Likewise, silencing of Wt1 in a mouse mesonephros-derived cell line reduced Igfbp5 mRNA levels by approximately 80%. Conversely, induction of the WT1(-KTS) isoform, whose role as transcriptional regulator has been firmly established, significantly increased IGFBP5 mRNA and protein levels in osteosarcoma cells. IGFBP5 expression was not significantly changed by WT1(+KTS) protein, which exhibits lower DNA binding affinity than the WT1(-KTS) isoform and has a presumed role in post-transcriptional gene regulation. Luciferase reporter constructs harboring 0.8 and 1.6 kilobases of the murine Igfbp5 promoter, respectively, were stimulated approximately 5-fold by co-transfection of WT1(-KTS). The WT1(+KTS) variant had no significant effect on IGFBP5 promoter activity. Binding of WT1(-KTS), but not of WT1(+KTS) protein, to the IGFBP5 promoter in human osteosarcoma cells was proven by chromatin immunoprecipitation (ChIP) and confirmed by electrophoretic mobility shift assay. These findings demonstrate that WT1 activates transcription of the IGFBP5 gene with possible implications for kidney development and bone (patho)physiology. Copyright © 2017 Elsevier B.V. All rights reserved.

Interconnection of thermal parameters, microstructure and mechanical properties in directionally solidified Sn–Sb lead-free solder alloys

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dias, Marcelino; Costa, Thiago; Rocha, Otávio

2015-08-15

Considerable effort is being made to develop lead-free solders for assembling in environmental-conscious electronics, due to the inherent toxicity of Pb. The search for substitute alloys of Pb–Sn solders has increased in order to comply with different soldering purposes. The solder must not only meet the expected levels of electrical performance but may also have appropriate mechanical strength, with the absence of cracks in the solder joints. The Sn–Sb alloy system has a range of compositions that can be potentially included in the class of high temperature solders. This study aims to establish interrelations of solidification thermal parameters, microstructure andmore » mechanical properties of Sn–Sb alloys (2 wt.%Sb and 5.5 wt.%Sb) samples, which were directionally solidified under cooling rates similar to those of reflow procedures in industrial practice. A complete high-cooling rate cellular growth is shown to be associated with the Sn–2.0 wt.%Sb alloy and a reverse dendrite-to-cell transition is observed for the Sn–5.5 wt.%Sb alloy. Strength and ductility of the Sn–2.0 wt.%Sb alloy are shown not to be affected by the cellular spacing. On the other hand, a considerable variation in these properties is associated with the cellular region of the Sn–5.5 wt.%Sb alloy casting. - Graphical abstract: Display Omitted - Highlights: • The microstructure of the Sn–2 wt.%Sb alloy is characterized by high-cooling rates cells. • Reverse dendrite > cell transition occurs for Sn–5.5 wt.%Sb alloy: cells prevail for cooling rates > 1.2 K/s. • Sn–5.5 wt.%Sb alloy: the dendritic region occurs for cooling rates < 0.9 K/s. • Sn–5.5 wt.%Sb alloy: tensile properties are improved with decreasing cellular spacing.« less

Monitoring fluidized bed drying of pharmaceutical granules.

PubMed

Briens, Lauren; Bojarra, Megan

2010-12-01

Placebo granules consisting of lactose monohydrate, corn starch, and polyvinylpyrrolidone were prepared using de-ionized water in a high-shear mixer and dried in a conical fluidized bed dryer at various superficial gas velocities. Acoustic, vibration, and pressure data obtained over the course of drying was analyzed using various statistical, frequency, fractal, and chaos techniques. Traditional monitoring methods were also used for reference. Analysis of the vibration data showed that the acceleration levels decreased during drying and reached a plateau once the granules had reached a final moisture content of 1–2 wt.%; this plateau did not differ significantly between superficial gas velocities, indicating a potential criterion to support drying endpoint identification. Acoustic emissions could not reliably identify the drying endpoint. However, high kurtosis values of acoustic emissions measured in the filtered air exhaust corresponded to high entrainment rates. This could be used for process control to adjust the fluidization gas velocity to allow drying to continue rapidly while minimizing entrainment and possible product losses.

Treatment-Induced Autophagy Associated with Tumor Dormancy and Relapse

DTIC Science & Technology

2017-07-01

immunogenic apoptosis, autophagy and senescence in MMC and SKBR3 tumor cell lines. We determined that ADR, but not radiation , induced what appears to...hours after WT and BPTF KD 4T1 cells were treated with 5 Gy γ- radiation exposure. (D) γH2AX was measured by flow cytometry 30 minutes after WT and...BPTF KD 4T1 cells were treated with 6Gy γ- radiation . (E) Flow cytometry measurement of 1 ug/ml ADR accumulation after 24 hours exposure to WT and BPTF

Mercury in alligators (Alligator mississippiensis) in the southeastern United States

USGS Publications Warehouse

Jagoe, C.H.; Arnold-Hill, B.; Yanochko, G.M.; Winger, P.V.; Brisbin, I.L.

1998-01-01

Mercury methylation may be enhanced in wetlands and humic-rich, blackwater systems that crocodiles and alligators typically inhabit. Given their high trophic level and long life-spans, crocodilians could accumulate significant burdens of Hg. Our objectives were to survey Hg concentrations in alligators from several areas in the southeastern United States to test their utility as sentinels of Hg contamination, to examine relationships among Hg concentrations in various tissues and to look for any differences in tissue Hg concentrations among locations. We measured total Hg concentrations in alligators collected in the Florida Everglades (n = 18), the Okefenokee National Wildlife Refuge, Georgia (n = 9), the Savannah River Site (SRS), South Carolina (n = 49) and various locations in central Florida ( n = 21), sampling tissues including blood, brain, liver, kidney, muscle, bone, fat, spleen, claws and dermal scutes. Alligators from the Everglades were mostly juvenile, but Hg concentrations in tissues were high (means: liver 41.0, kidney 36.4, muscle 5.6 mg Hg/kg dry wt.). Concentrations in alligators from other locations in Florida were lower (means: liver 14.6, kidney 12.6, muscle 1.8 mg Hg/kg dry wt.), although they tended to be larger adults. Alligators from the Okefenokee were smallest and had the lowest Hg concentrations (means: liver 4.3, kidney 4.8, muscle 0.8 mg Hg/kg dry wt.). At some locations, alligator length was correlated with Hg concentrations in some internal organs. However, at three of the four locations, muscle Hg was not related to length. Tissue Hg concentrations were correlated at most locations; however, claw or dermal scute Hg explained less than 74% of the variation of Hg in muscle or organs, suggesting readily-obtained tissues, such as scutes or claws, have limited value for nondestructive screening of Hg in alligator populations.

Chloromethane Degradation in Soils: A Combined Microbial and Two-Dimensional Stable Isotope Approach.

PubMed

Jaeger, Nicole; Besaury, Ludovic; Kröber, Eileen; Delort, Anne-Marie; Greule, Markus; Lenhart, Katharina; Nadalig, Thierry; Vuilleumier, Stéphane; Amato, Pierre; Kolb, Steffen; Bringel, Françoise; Keppler, Frank

2018-03-01

Chloromethane (CHCl, methyl chloride) is the most abundant volatile halocarbon in the atmosphere and involved in stratospheric ozone depletion. The global CHCl budget, and especially the CHCl sink from microbial degradation in soil, still involves large uncertainties. These may potentially be resolved by a combination of stable isotope analysis and bacterial diversity studies. We determined the stable isotope fractionation of CHCl hydrogen and carbon and investigated bacterial diversity during CHCl degradation in three soils with different properties (forest, grassland, and agricultural soils) and at different temperatures and headspace mixing ratios of CHCl. The extent of chloromethane degradation decreased in the order forest > grassland > agricultural soil. Rates ranged from 0.7 to 2.5 μg g dry wt. d for forest soil, from 0.1 to 0.9 μg g dry wt. d for grassland soil, and from 0.1 to 0.4 μg g dry wt. d for agricultural soil and increased with increasing temperature and CHCl supplementation. The measured mean stable hydrogen enrichment factor of CHCl of -50 ± 13‰ was unaffected by temperature, mixing ratio, or soil type. In contrast, the stable carbon enrichment factor depended on CHCl degradation rates and ranged from -38 to -11‰. Bacterial community composition correlated with soil properties was independent from CHCl degradation or isotope enrichment. Nevertheless, increased abundance after CHCl incubation was observed in 21 bacterial operational taxonomical units (OTUs at the 97% 16S RNA sequence identity level). This suggests that some of these bacterial taxa, although not previously associated with CHCl degradation, may play a role in the microbial CHCl sink in soil. Copyright © by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America, Inc.

Bio sorption of strontium from aqueous solution by New Strain Bacillus sp. GTG-83

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tajer Mohammad Ghazvini, P.; Ghorbanzadeh Mashkani, S.; Ghafourian, H.

Attempt was made to isolate bacterial strains capable of removing Sr biologically. In this study we collected ten different water samples from naturally radioactive spring Neydasht in Iran and bacterial strains samples isolated. Initial screening of a total of 50 bacterial isolates resulted in selection of one strain. The strain showed maximum adsorption capacity with 55 mg Sr/g dry wt. It was tentatively identified as Bacillus sp. according to morphological and biochemical properties and called strain GTG-83. Studies indicated that Bacillus sp. GTG-83 was able to grow aerobically in the presence of 50 mM SrCl{sub 2} but showed severe growthmore » inhibition at levels above that concentration. The bio-sorption capacity of Bacillus sp. GTG-83 strongly depends on solution pH, and the maximum Sr sorption capacity of Bacillus sp. GTG-83 were obtained at pH 10 independent of the absence or the presence of increasing concentrations of salt (MgCl{sub 2}). Sr-salt bio-sorption studies were also performed at this pH values. Equilibrium uptakes of Sr increased with increasing Sr concentrations up to 250 mg/l for Bacillus sp. GTG-83. Maximum bio-sorption of Sr was obtained at temperatures in the range of 30-35 deg. C. Bacillus sp. GTG-83 bio-sorbed 97 mg Sr/g dry wt at 100 mg/l initial Sr concentration without salt medium (MgCl{sub 2}). When salt concentration (MgCl{sub 2}) increased to 15% (w/v), these values dropped to 23.6 mg Sr/g dry wt at the same conditions. Uptake of Sr within 5 min of incubation was relatively rapid and the absorption continued slowly thereafter. (authors)« less

Major correlates of mercury in small fish and common loons (Gavia immer) across four large study areas in Canada.

PubMed

Scheuhammer, A M; Lord, S I; Wayland, M; Burgess, N M; Champoux, L; Elliott, J E

2016-03-01

We investigated mercury (Hg) concentrations in small fish (mainly yellow perch, Perca flavescens; ∼60% of fish collected) and in blood of common loons (Gavia immer) that prey upon them during the breeding season on lakes in 4 large, widely separated study areas in Canada (>13 lakes per study area; total number of lakes = 93). Although surface sediments from lakes near a base metal smelter in Flin Flon, Manitoba had the highest Hg concentrations, perch and other small fish and blood of common loon chicks sampled from these same lakes had low Hg concentrations similar to those from uncontaminated reference lakes. Multiple regression modeling with AIC analysis indicated that lake pH was by far the most important single factor influencing perch Hg concentrations in lakes across the four study areas (R(2) = 0.29). The best model was a three-variable model (pH + alkalinity + sediment Se; Wi = 0.61, R(2) = 0.85). A single-variable model (fish Hg) best explained among-lake variability in loon chick blood Hg (Wi = 0.17; R(2) = 0.53). From a toxicological risk perspective, all lakes posing a potential Hg health risk for perch and possibly other small pelagic fish species (where mean fish muscle Hg concentrations exceeded 2.4 μg/g dry wt.), and for breeding common loons (where mean fish muscle Hg concentrations exceeded 0.8 μg/g dry wt., and loon chick blood Hg exceeded 1.4 μg/g dry wt.) had pH < 6.7 and were located in eastern Canada. Crown Copyright © 2016. Published by Elsevier Ltd. All rights reserved.

Impact of T-cell-specific Smad4 deficiency on the development of autoimmune diabetes in NOD mice

PubMed Central

Kim, Donghee; Lee, Song Mi; Jun, Hee-Sook

2017-01-01

Type 1 diabetes results from autoimmune-mediated pancreatic beta-cell destruction and transforming growth factor-beta (TGF-β) is known to play a preventive role in type 1 diabetes in non-obese diabetic (NOD) mice. In this study, we investigated the role of Smad4, a key molecule for Smad-dependent TGF-β signaling, in T cells of NOD mice in the pathogenesis of autoimmune diabetes. We generated T-cell-specific Smad4 knockout (Smad4 tKO) NOD mice and assessed the pathological and immunological changes. Smad4 tKO showed earlier onset and increased incidence of diabetes than wild type (WT) NOD mice. Pathological features such as insulitis, anti-glutamic acid decarboxylase auto-antibody levels and serum IFN-γ levels were significantly increased in Smad4 tKO compared with WT NOD mice. Proportion and number of activated/memory CD4+ T cell were significantly increased in pancreatic lymph nodes of Smad4 tKO compared with WT NOD mice. However, the proportion and function of regulatory T cells was not different. Effector CD4+ T cells from Smad4 tKO were more resistant to suppression by regulatory T cells than effector cells from WT NOD mice. The proliferative potential of effector T cells from Smad4 tKO was significantly elevated compared with WT NOD mice, and activation of sterol regulatory element binding protein-1c (SREBP-1c) in T cells of Smad4 tKO NOD mice was correlated with this proliferative activity. We conclude that Smad4 deletion in T cells of NOD mice accelerated the development of autoimmune diabetes and increased the incidence of the disease by dysregulation of T cell activation at least in part via SREBP-1c activation. PMID:27686408

Impact of T-cell-specific Smad4 deficiency on the development of autoimmune diabetes in NOD mice.

PubMed

Kim, Donghee; Lee, Song Mi; Jun, Hee-Sook

2017-03-01

Type 1 diabetes results from autoimmune-mediated pancreatic beta-cell destruction and transforming growth factor-beta (TGF-β) is known to play a preventive role in type 1 diabetes in non-obese diabetic (NOD) mice. In this study, we investigated the role of Smad4, a key molecule for Smad-dependent TGF-β signaling, in T cells of NOD mice in the pathogenesis of autoimmune diabetes. We generated T-cell-specific Smad4 knockout (Smad4 tKO) NOD mice and assessed the pathological and immunological changes. Smad4 tKO showed earlier onset and increased incidence of diabetes than wild type (WT) NOD mice. Pathological features such as insulitis, anti-glutamic acid decarboxylase auto-antibody levels and serum IFN-γ levels were significantly increased in Smad4 tKO compared with WT NOD mice. Proportion and number of activated/memory CD4 + T cell were significantly increased in pancreatic lymph nodes of Smad4 tKO compared with WT NOD mice. However, the proportion and function of regulatory T cells was not different. Effector CD4 + T cells from Smad4 tKO were more resistant to suppression by regulatory T cells than effector cells from WT NOD mice. The proliferative potential of effector T cells from Smad4 tKO was significantly elevated compared with WT NOD mice, and activation of sterol regulatory element binding protein-1c (SREBP-1c) in T cells of Smad4 tKO NOD mice was correlated with this proliferative activity. We conclude that Smad4 deletion in T cells of NOD mice accelerated the development of autoimmune diabetes and increased the incidence of the disease by dysregulation of T cell activation at least in part via SREBP-1c activation.

Sjögren's syndrome associated dry eye in a mouse model is ameliorated by topical application of integrin α4 antagonist GW559090.

PubMed

Contreras-Ruiz, Laura; Mir, Fayaz A; Turpie, Bruce; Krauss, Achim H; Masli, Sharmila

2016-02-01

Sjögren's syndrome is an autoimmune disease associated with inflammation of exocrine glands with clinical manifestations of dry eye and dry mouth. Dry eye in this disease involves inflammation of the ocular surface tissues - cornea and conjunctiva. While systemic blockade of adhesion molecules has been used to treat autoimmune diseases, the purpose of this study was to determine the therapeutic efficacy of topical application of an integrin α4 adhesion molecule antagonist in a mouse model of dry eye associated with Sjögren's syndrome. To assess this spontaneously developed ocular surface inflammation related to Sjögren's syndrome in TSP-1null mice (12 wks) was evaluated. Mice were treated with topical formulations containing 0.1% dexamethasone or 30 mg/ml GW559090 or vehicle control. Corneal fluorescein staining and conjunctival goblet cell density were assessed. Real-time PCR analysis was performed to assess expression of the inflammatory marker IL-1β in the cornea and Tbet and RORγt in the draining lymph nodes. Ocular surface inflammation was detectable in TSP-1null mice (≥12 wk old), which resulted in increased corneal fluorescein staining indicative of corneal barrier disruption and reduced conjunctival goblet cell density. These changes were accompanied by increased corneal expression of IL-1β as compared to WT controls and an altered balance of Th1 (Tbet) and Th17 (RORγt) markers in the draining lymph nodes. Topically applied dexamethasone and GW559090 significantly reduced corneal fluorescein staining compared to vehicle treatment (p = 0.023 and p < 0.001, respectively). This improved corneal barrier integrity upon adhesion molecule blockade was consistent with significantly reduced corneal expression of pro-inflammatory IL-1β compared to vehicle treated groups (p < 0.05 for both treatments). Significant improvement in goblet cell density was also noted in mice treated with 0.1% dexamethasone and GW559090 (p < 0.05 for both). We conclude that similar to topical dexamethasone, topically administered GW559090 successfully improved corneal barrier integrity and inflammation in an established ocular surface disease associated with Sjögren's syndrome. Copyright © 2015 Elsevier Ltd. All rights reserved.

Mechanical properties and shape memory effect of thermal-responsive polymer based on PVA

NASA Astrophysics Data System (ADS)

Lin, Liulan; Zhang, Lingfeng; Guo, Yanwei

2018-01-01

In this study, the effect of content of glutaraldehyde (GA) on the shape memory behavior of a shape memory polymer based on polyvinyl alcohol chemically cross-linked with GA was investigated. Thermal-responsive shape memory composites with three different GA levels, GA-PVA (3 wt%, 5 wt%, 7 wt%), were prepared by particle melting, mold forming and freeze-drying technique. The mechanical properties, thermal properties and shape memory behavior were measured by differential scanning calorimeter, physical bending test and cyclic thermo-mechanical test. The addition of GA to PVA led to a steady shape memory transition temperature and an improved mechanical compressive strength. The composite with 5 wt% of GA exhibited the best shape recoverability. Further increase in the crosslinking agent content of GA would reduce the recovery force and prolong the recovery time due to restriction in the movement of the soft PVA chain segments. These results provide important information for the study on materials in 4D printing.

Galectin-3 disruption impaired tumoral angiogenesis by reducing VEGF secretion from TGFβ1-induced macrophages.

PubMed

Machado, Camila Maria Longo; Andrade, Luciana Nogueira Sousa; Teixeira, Verônica Rodrigues; Costa, Fabrício Falconi; Melo, Camila Morais; dos Santos, Sofia Nascimento; Nonogaki, Suely; Liu, Fu-Tong; Bernardes, Emerson Soares; Camargo, Anamaria Aranha; Chammas, Roger

2014-04-01

In order to study the role of galectin-3 in tumor angiogenesis associated with tumor-associated macrophages (TAM) and tumor parenchyma, the galectin-3 expression was reconstituted in Tm1 melanoma cell line that lacks this protein. Galectin-3-expressing cells (Tm1G3) and mock-vector transfected cells (Tm1N3) were injected into wild-type (WT) and galectin-3 knockout (KO) C57Bl/6 mice. Tumors originated from Tm1G3 were larger in tumor volume with enlarged functional vessels, decreased necrotic areas, and increased vascular endothelial growth factor (VEGF) protein levels. Galectin-3-nonexpressing-cells injected into WT and KO showed increased levels of transforming growth factor beta 1 (TGFβ1) and, in WT animals this feature was also accompanied by increased VEGFR2 expression and its phosphorylation. In KO animals, tumors derived from galectin-3-expressing cells were infiltrated by CD68(+)-cells, whereas in tumors derived from galectin-3-nonexpressing-cells, CD68(+) cells failed to infiltrate tumors and accumulated in the periphery of the tumor mass. In vitro studies showed that Tm1G3 secreted more VEGF than Tm1N3 cells. In the latter case, TGFβ1 induced VEGF production. Basal secretion of VEGF was higher in WT-bone marrow-derived macrophages (BMDM) than in KO-BMDM. TGFβ1 induced secretion of VEGF only in WT-BMDM. Tm1G3-induced tumors had the Arginase I mRNA increased, which upregulated alternative macrophage (M2)/TAM induction. M2 stimuli, such as interleukin-4 (IL4) and TGFβ1, increased Arginase I protein levels and galectin-3 expression in WT- BMDM, but not in cells from KO mice. Hence, we report that galectin-3 disruption in tumor stroma and parenchyma decreases angiogenesis through interfering with the responses of macrophages to the interdependent VEGF and TGFβ1 signaling pathways. © 2014 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.

Alphavirus production is inhibited in neurofibromin 1-deficient cells through activated RAS signalling

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kolokoltsova, Olga A.; Domina, Aaron M.; Kolokoltsov, Andrey A.

2008-07-20

Virus-host interactions essential for alphavirus pathogenesis are poorly understood. To address this shortcoming, we coupled retrovirus insertional mutagenesis and a cell survival selection strategy to generate clonal cell lines broadly resistant to Sindbis virus (SINV) and other alphaviruses. Resistant cells had significantly impaired SINV production relative to wild-type (WT) cells, although virus binding and fusion events were similar in both sets of cells. Analysis of the retroviral integration sites identified the neurofibromin 1 (NF1) gene as disrupted in alphavirus-resistant cell lines. Subsequent analysis indicated that expression of NF1 was significantly reduced in alphavirus-resistant cells. Importantly, independent down-regulation of NF1 expressionmore » in WT HEK 293 cells decreased virus production and increased cell viability during SINV infection, relative to infected WT cells. Additionally, we observed hyperactive RAS signalling in the resistant HEK 293 cells, which was anticipated because NF1 is a negative regulator of RAS. Expression of constitutively active RAS (HRAS-G12V) in a WT HEK 293 cell line resulted in a marked delay in virus production, compared with infected cells transfected with parental plasmid or dominant-negative RAS (HRAS-S17N). This work highlights novel host cell determinants required for alphavirus pathogenesis and suggests that RAS signalling may play an important role in neuronal susceptibility to SINV infection.« less

The microstructural dependence of wear resistance in austenite containing plate steels

NASA Astrophysics Data System (ADS)

Wolfram, Preston Charles

The purpose of this project was to examine the microstructural dependence of wear resistance of various plate steels, with interests in exploring the influence of retained austenite (RA). Materials resistant to abrasive wear are desirable in the industrial areas of agriculture, earth moving, excavation, mining, mineral processing, and transportation. Abrasive wear contributes to significant financial cost associated with wear to the industry. The motivation for the current study was to determine whether it would be beneficial from a wear resistance perspective to produce plate steels with increased amounts of retained austenite. This thesis investigates this motivation through a material matrix containing AR400F, Abrasive (0.21 wt pct C, 1.26 wt pct Mn, 0.21 wt pct Si, 0.15 wt pct Ni, 0.18 wt pct Mo), Armor (0.46 wt pct C, 0.54 wt pct Mn, 0.36 wt pct Si, 1.74 wt pct Ni, 0.31 wt pct Mo), 9260, 301SS, Hadfield, and SAE 4325 steels. The Abrasive, Armor and 9260 steels were heat treated using different methods such as quench and temper, isothermal bainitic hold, and quench and partitioning (Q&P). These heat treatments yielded various microstructures and the test matrix allowed for investigation of steels with similar hardness and varying levels of RA. The wear test methods used consisted of dry sand rubber wheel (DSRW), impeller-tumbler impact-abrasion (impeller), and Bond abrasion wear testing. DSRW and impeller wear resistance was found to increase with hardness and retained austenite levels at certain hardness levels. Some Q&P samples exhibited similar or less wear than the Hadfield steels in DSRW and impeller tests. Scanning electron microscopy investigation of wear surfaces revealed different wear mechanisms for the different wear test methods ranging from micro-plowing, to micro-cutting and to fragmentation.

Expression of the Wilms' tumor gene WT1 in the murine urogenital system.

PubMed

Pelletier, J; Schalling, M; Buckler, A J; Rogers, A; Haber, D A; Housman, D

1991-08-01

The Wilms' tumor gene WT1 is a recessive oncogene that encodes a putative transcription factor implicated in nephrogenesis during kidney development. In this report we analyze expression of WT1 in the murine urogenital system. WT1 is expressed in non-germ-cell components of the testis and ovaries in both young and adult mice. In situ mRNA hybridization studies demonstrate that WT1 is expressed in the granulosa and epithelial cells of ovaries, the Sertoli cells of the testis, and in the uterine wall. In addition to the 3.1-kb WT1 transcript detected by Northern blotting of RNA from kidney, uterus, and gonads, there is an approximately 2.5-kb WT1-related mRNA species in testis. The levels of WT1 mRNA in the gonads are among the highest observed, surpassing amounts detected in the embryonic kidney. During development, these levels are differentially regulated, depending on the sexual differentiation of the gonad. Expression of WT1 mRNA in the female reproductive system does not fluctuate significantly from days 4 to 40 postpartum. In contrast, WT1 mRNA levels in the tesis increase steadily after birth, reaching their highest expression levels at day 8 postpartum and decreasing slightly as the animal matures. Expression of WT1 in the gonads is detectable as early as 12.5 days postcoitum (p.c.). As an initial step toward exploring the tissue-specific expression of WT1, DNA elements upstream of WT1 were cloned and sequenced. Three putative transcription initiation sites, utilized in testis, ovaries, and uterus, were mapped by S1 nuclease protection assays. The sequences surrounding these sites have a high G + C content, and typical upstream CCAAT and TATAA boxes are not present. These studies allowed us to identify the translation initiation site for WT1 protein synthesis. We have also used an epitope-tagging protocol to demonstrate that WT1 is a nuclear protein, consistent with its role as a transcription factor. Our results demonstrate regulation of WT1 expression during development of the gonads, implicate WT1 in genitourinary development, and provide a molecular framework toward understanding genitourinary defects observed among hereditary cases of Wilms' tumor.

IL-23 is critical in the induction but not in the effector phase of experimental autoimmune encephalomyelitis.

PubMed

Thakker, Paresh; Leach, Michael W; Kuang, Wen; Benoit, Stephen E; Leonard, John P; Marusic, Suzana

2007-02-15

Experimental autoimmune encephalomyelitis (EAE), a T cell-mediated inflammatory disease of the CNS, is a rodent model of human multiple sclerosis. IL-23 is one of the critical cytokines in EAE development and is currently believed to be involved in the maintenance of encephalitogenic responses during the tissue damage effector phase of the disease. In this study, we show that encephalitogenic T cells from myelin oligodendrocyte glycopeptide (MOG)-immunized wild-type (WT) mice caused indistinguishable disease when adoptively transferred to WT or IL-23-deficient (p19 knockout (KO)) recipient mice, demonstrating that once encephalitogenic cells have been generated, EAE can develop in the complete absence of IL-23. Furthermore, IL-12/23 double-deficient (p35/p19 double KO) recipient mice developed EAE that was indistinguishable from WT recipients, indicating that IL-12 did not compensate for IL-23 deficiency during the effector phase of EAE. In contrast, MOG-specific T cells from p19KO mice induced EAE with delayed onset and much lower severity when transferred to WT recipient mice as compared with the EAE that was induced by cells from WT controls. MOG-specific T cells from p19KO mice were highly deficient in the production of IFN-gamma, IL-17A, and TNF, indicating that IL-23 plays a critical role in development of encephalitogenic T cells and facilitates the development of T cells toward both Th1 and Th17 pathways.

Effect of particle size and ammonium sulfate concentration on rice bran fermentation with the fungus Rhizopus oryzae.

PubMed

Schmidt, Cristiano Gautério; Furlong, Eliana Badiale

2012-11-01

The effects of rice bran particle size (0.18-0.39mm) and ammonium sulfate concentration in the nutrient solution (2-8g/L) on biomass production, protein and phenolic content generated by solid state fermentation with the fungus Rhizopus oryzae (CCT 1217) were studied. Particle size had a positive effect on biomass production and a negative effect (p⩽0.05) on protein and phenolic contents. Ammonium sulfate concentration had a positive effect (p⩽0.05) on biomass and phenolic content gain. Cultivation of fungus in rice bran with particle size of 0.18mm and in the presence of 8g/L ammonium sulfate, resulted in protein levels of 20g/100g dry wt and phenolics content of 4mg/g dry wt. These values were 53 and 65% higher than those achieved with unfermented rice bran. The results demonstrate that the fermentation process increased the value of compounds recovered for potential use in food formulations. Copyright © 2012 Elsevier Ltd. All rights reserved.

Distribution and accumulation of organotin species in seawater, sediments and organisms collected from a Taiwan mariculture area.

PubMed

Liu, Li-Lian; Wang, Jih-Terng; Chung, Kuo-Nan; Leu, Ming-Yih; Meng, Pei-Jie

2011-01-01

The present study was undertaken to evaluate the distribution and accumulation of tributyltin (TBT) and triphenyltin (TPhT) in seawater, sediments and selected organisms from a cage mariculture area in southern Taiwan, Hsiao Liouchiou Island. Our results show that ΣOTs were found in concentrations as high as 196 ng/L in seawater collected from the sites in Pai-Sa harbor, and up 1040 ng/g dry wt. in sediments dredged from sites within Da-Fu harbor. Also, ΣOTs concentrations of 859 ng/g dry wt. were observed in the liver of cobia (Rachycentron canadum) from mariculture cages. As most published studies have focused on the acute toxicity and bioaccumulation of organotins in mussels, the effects of organotins on cobia and other marine fauna are still poorly understood. This study highlights the significance of ΣBTs accumulation in cobia, as well as in the sediments and seawater surrounding their culture facilities. Copyright © 2011 Elsevier Ltd. All rights reserved.

The Effect of Nanocopper Additions in a Urea-Formaldehyde Adhesive on the Physical and Mechanical Properties of Particleboard Manufactured from Date Palm Waste

NASA Astrophysics Data System (ADS)

Rangavar, H.; Hoseiny fard, M. S.

2015-03-01

The effect of addition of copper nanoparticles to a urea-formaldehyde (UF) adhesive on the physical and mechanical properties of particleboards manufactured from date palm waste (DPW) was investigated. The variable factors in the study included copper nanoparticles in amounts of 6 and 8 wt.% of the dry mass of wood, pressing durations of 5 and 6 min, and pressing temperatures of 150 and 160°C. The physical and mechanical properties of manufactured boards were measured according to EN standards. The results showed that the addition of copper nanoparticles to the UF adhesive considerably improved the physical and mechanical properties of the boards and shortened the pressing duration. The boards manufactured with 6 wt.% copper nanoparticles in a dry mass of wood mixed with the adhesive and pressed at a temperature of 160°C for 5 min had mechanical properties exceeding the EN312-2 standard levels.

Solvo-thermal in situ transesterification of wet spent coffee grounds for the production of biodiesel.

PubMed

Park, Jeongseok; Kim, Bora; Son, Jeesung; Lee, Jae W

2018-02-01

This work addresses non-catalytic biodiesel production from spent coffee ground (SCG) by integrating solvo-thermal effect of 1,2-dichloroethane (DCE) with in situ transesterification over 160 °C. The SCG water content has a positive effect on the DCE hydrolysis up to 60 wt% due to the bimolecular substitution mechanism. The hydrolysis gives an acidic environment favorable for cellulose decomposition, SCG particle size reduction and lipid conversion. The optimal fatty acid ethyl ester yield was 11.8 wt% based on the mass of dried SCG with 3.36 ml ethanol and 3.16 ml DCE at 196.8 °C through the response surface methodology. Using the solvo-thermal effect, direct utilization of wet SCG as a biodiesel feedstock provides not only economic feasibility without using drying process and additional acid catalyst but also environmental advantage of recycling the municipal waste. Copyright © 2017 Elsevier Ltd. All rights reserved.

Production of cellulosic ethanol from cotton processing residues after pretreatment with dilute sodium hydroxide and enzymatic hydrolysis.

PubMed

Fockink, Douglas Henrique; Maceno, Marcelo Adriano Corrêa; Ramos, Luiz Pereira

2015-01-01

In this study, production of cellulosic ethanol from two cotton processing residues was investigated after pretreatment with dilute sodium hydroxide. Pretreatment performance was investigated using a 2(2) factorial design and the highest glucan conversion was achieved at the most severe alkaline conditions (0.4g NaOH g(-1) of dry biomass and 120°C), reaching 51.6% and 38.8% for cotton gin waste (CGW) and cotton gin dust (CGD), respectively. The susceptibility of pretreated substrates to enzymatic hydrolysis was also investigated and the best condition was achieved at the lowest total solids (5wt%) and the highest enzyme loading (85mg of Cellic CTec2 g(-1) of dry substrate). However, the highest concentration of fermentable sugars - 47.8 and 42.5gL(-1) for CGD and CGW, respectively - was obtained at 15wt% total solids using this same enzyme loading. Substrate hydrolysates had no inhibitory effects on the fermenting microorganism. Copyright © 2015. Published by Elsevier Ltd.

Activated leucocyte cell adhesion molecule (ALCAM/CD166) regulates T cell responses in a murine model of food allergy.

PubMed

Kim, Y S; Kim, M N; Lee, K E; Hong, J Y; Oh, M S; Kim, S Y; Kim, K W; Sohn, M H

2018-05-01

Food allergy is a major public health problem. Studies have shown that long-term interactions between activated leucocyte cell adhesion molecule (ALCAM/CD166) on the surface of antigen-presenting cells, and CD6, a co-stimulatory molecule, influence immune responses. However, there are currently no studies on the functions of ALCAM in food allergy. Therefore, we aimed to identify the functions of ALCAM in ovalbumin (OVA)-induced food allergy using ALCAM-deficient mice. Wild-type (WT) and ALCAM-deficient (ALCAM -/- ) mice were sensitized intraperitoneally and with orally fed OVA. The mice were killed, and parameters related to food allergy and T helper type 2 (Th2) immune responses were analysed. ALCAM serum levels increased and mRNA expression decreased in OVA-challenged WT mice. Serum immunoglobulin (Ig)E levels, Th2 cytokine mRNA and histological injuries were higher in OVA-challenged WT mice than in control mice, and these were attenuated in ALCAM -/- mice. T cell proliferation of total cells, CD3 + CD4 + T cells and activated T cells in immune tissues were diminished in OVA-challenged ALCAM -/- mice. Proliferation of co-cultured T cells and dendritic cells (DCs) was decreased by the anti-CD6 antibody. In addition, WT mice sensitized by adoptive transfer of OVA-pulsed ALCAM -/- BM-derived DCs showed reduced immune responses. Lastly, serum ALCAM levels were higher in children with food allergy than in control subjects. In this study, serum levels of ALCAM were elevated in food allergy-induced WT mice and children with food allergy. Moreover, immune responses and T cell activation were attenuated in OVA-challenged ALCAM -/- mice. These results indicate that ALCAM regulates food allergy by affecting T cell activation. © 2018 British Society for Immunology.

EFFECT OF THAI SARAPHI FLOWER EXTRACTS ON WT1 AND BCR/ABL PROTEIN EXPRESSION IN LEUKEMIC CELL LINES.

PubMed

Sangkaruk, Rungkarn; Rungrojsakul, Methee; Tima, Singkome; Anuchapreeda, Songyot

2017-01-01

Saraphi (Mammea siamensis) is a Thai traditional herb. In this study, the cytotoxic effects of crude ethanolic and fractional extracts including hexane, ethyl acetate, and methanol fractions from M. siamensis flowers were investigated in order to determine their effect on WT1 expression in Molt4 and K562 cells and Bcr/Abl expression in K562 cells. The flowers of M. siamensis were extracted using ethanol. The ethanol flower extract was further fractionated with hexane, ethyl acetate, and methanol. Cytotoxic effects were measured by the MTT assay. Bcr/Abl and WT1 protein levels after treatments were determined by Western blotting. The total cell number was determined via the typan blue exclusion method. The hexane fraction showed the strongest cytotoxic activity on Molt4 and K562 cells, with IC 50 values of 2.6 and 77.6 μg/ml, respectively. The hexane extract decreased Bcr/Abl protein expression in K562 cells by 74.6% and WT1 protein expressions in Molt4 and K562 cells by 68.4 and 72.1%, respectively. Total cell numbers were decreased by 66.2 and 48.7% in Molt4 and K562 cells, respectively. Mammea E/BB (main active compound) significantly decreased both Bcr/Abl and WTlprotein expressions by 75 and 49.5%, respectively when compared to vehicle control. The hexane fraction from M. siamensis flowers inhibited cell proliferation via the suppression of WT1 expression in Molt4 and K562 cells and Bcr/Abl expression in K562 cells. The active compound may be mammea E/BB. Extracts from M. siamensis flowers show promise as naturally occurring anti-cancer drugs.

Phosphorylation of interleukin (IL)-24 is required for mediating its anti-cancer activity.

PubMed

Panneerselvam, Janani; Shanker, Manish; Jin, Jiankang; Branch, Cynthia D; Muralidharan, Ranganayaki; Zhao, Yan D; Chada, Sunil; Munshi, Anupama; Ramesh, Rajagopal

2015-06-30

Interleukin (IL)-24 is a tumor suppressor/cytokine gene that undergoes post-translational modifications (PTMs). Glycosylation and ubiquitination are important for IL-24 protein stabilization and degradation respectively. Little is known about IL-24 protein phosphorylation and its role in IL-24-mediated anti-tumor activities. In this study we conducted molecular studies to determine whether IL-24 phosphorylation is important for IL-24-mediated anti-cancer activity.Human H1299 lung tumor cell line that was stably transfected with a doxycycline (DOX)-inducible (Tet-on) plasmid vector carrying the cDNA of IL-24-wild-type (IL-24wt) or IL-24 with all five phosphorylation sites replaced (IL-24mt) was used in the present study. Inhibition of tumor cell proliferation, cell migration and invasion, and induction of G2/M cell cycle arrest was observed in DOX-induced IL-24wt-expressing cells but not in IL-24mt-expressing cells. Secretion of IL-24mt protein was greatly reduced compared to IL-24wt protein. Further, IL-24wt and IL-24mt proteins markedly differed in their subcellular organelle localization. IL-24wt but not IL-24mt inhibited the AKT/mTOR signaling pathway. SiRNA-mediated AKT knockdown and overexpression of myristolyated AKT protein confirmed that IL-24wt but not IL-24mt mediated its anti-cancer activity by inhibiting the AKT signaling pathway.Our results demonstrate that IL-24 phosphorylation is required for inhibiting the AKT/mTOR signaling pathway and exerting its anti-cancer activities.

Dry cell battery poisoning

MedlinePlus

Batteries - dry cell ... Acidic dry cell batteries contain: Manganese dioxide Ammonium chloride Alkaline dry cell batteries contain: Sodium hydroxide Potassium hydroxide Lithium dioxide dry cell batteries ...

Performance evaluation of GDC-SrMoO4-YSZ SOFCs prepared with different pore formers

NASA Astrophysics Data System (ADS)

Hongxin, You; Lian, Peng; Xiaojuan, Wang; Cong, Zhao; Yajun, Guan; Tao, Yu; Lijun, Xu; Abuliti

2018-04-01

The paper aims to evaluate the performance of anodes prepared with different pore formers. Anodic precursor material SrMoO4 was prepared by hard template method. Gd0.2Ce0.8O1.9 (GDC) was introduced to the precursor to prepare composite anode material GDC-SrMoO4-YSZ by wet impregnation method. Cotton-fibers, graphite powder, flour and activated carbon fibers (ACF) were added as pore formers to the anode to prepare the corresponding solid oxide fuel cell (SOFC), respectively. The electrical performance testing was conducted under the methane environment at 800°C. The result showed that the single cell with 5wt% cotton-fibers as anode pore-former performed best with the maximum power density (464.49 mW.cm2). The cross section samples of the test cells indicated that the anode was left with a plenty of continuous long channels because of the burning of cotton-fibers. Thus, the influence of the amount of cotton-fibers (2wt%, 4wt%, 5wt%, 7wt%, 10wt%) of the anode on the performance of SOFC was tested and further analyzed by the scanning electron microscope (SEM). It was indicated that the optimum adding amount of cotton-fibers was 5wt%.

Galectin-3 is essential for proper bone cell differentiation and activity, bone remodeling and biomechanical competence in mice.

PubMed

Iacobini, Carla; Fantauzzi, Claudia Blasetti; Bedini, Rossella; Pecci, Raffaella; Bartolazzi, Armando; Amadio, Bruno; Pesce, Carlo; Pugliese, Giuseppe; Menini, Stefano

2018-02-09

Galectin-3 is constitutively expressed in bone cells and was recently shown to modulate osteogenic transdifferentiation of vascular smooth muscle cells and atherosclerotic calcification. However, the role of galectin-3 in bone physiology is largely undefined. To address this issue, we analyzed (1) the skeletal features of 1-, 3- and 6-month-old galectin-3 null (Lgals3 -/- ) and wild type (WT) mice and (2) the differentiation and function of osteoblasts and osteoclasts derived from these animals. Long bone phenotype, gene expression profile, and remodeling were investigated by micro-computed tomography, real time-PCR, static and dynamic histomorphometry, and assessment of biochemical markers of bone resorption and formation. Bone competence was also evaluated by biomechanical testing at 3 months. In vitro, the effects of galectin-3 deficiency on bone cell differentiation and function were investigated by assessing (a) gene expression of osteoblast markers, alkaline phosphatase activity, mineralization assay, and WNT/β-catenin signaling (of which galectin-3 is a known regulator) in osteoblasts; and (b) tartrate-resistant acid phosphatase activity and bone resorption activity in osteoclasts. Lgals3 -/- mice revealed a wide range of age-dependent alterations including lower bone formation and higher bone resorption, accelerated age-dependent trabecular bone loss (p < 0.01 vs. WT at 3 months) and reduced bone strength (p < 0.01 vs. WT at 3 months). These abnormalities were accompanied by a steady inflammatory state, as revealed by higher bone expression of the pro-inflammatory cytokines interleukin (IL)-1β and IL-6 (p < 0.001 vs. WT at 3 months), increased content of osteal macrophages (p < 0.01 vs. WT at 3 months), and reduced expression of markers of alternative (M2) macrophage activation. Lgals3 -/- osteoblasts and osteoclasts showed impaired terminal differentiation, reduced mineralization capacity (p < 0.01 vs. WT cells) and resorption activity (p < 0.01 vs. WT cells). Mechanistically, impaired differentiation and function of Lgals3 -/- osteoblasts was associated with altered WNT/β-catenin signaling (p < 0.01 vs. WT cells). These data provide evidence for a contribution of galectin-3 to bone cell maturation and function, bone remodeling, and biomechanical competence, thus identifying galectin-3 as a promising therapeutic target for age-related disorders of bone remodeling. Copyright © 2018. Published by Elsevier Inc.

The role of the PI3K/mTOR signaling pathway in Staphylococcus epidermidis small colony variants intracellular survival.

PubMed

Magryś, Agnieszka; Bogut, Agnieszka; Kiełbus, Michał; Olender, Alina

2018-04-01

The objective of this study was to analyze how Staphylococcus epidermidis SCV and WT strains manipulate the PI3K/Akt/mTOR signaling pathway. Six S. epidermidis strains with normal phenotype (WT) and six S. epidermidis strains with SCV phenotype were isolated in parallel from six patients with the prosthetic hip joint infections. THP-1 activated cells were incubated with or without PI3K inhibitor-wortmannin or with mTOR inhibitor-rapamycin. Next, macrophages were exposed to S. epidermidis WT and SCV strains. After 4 h incubation, bacterial survival inside macrophages as well as PI3K-mTOR activation was analyzed. SCV strains of S. epidermidis increased the level of Akt phosphorylation, compared to uninfected macrophages and to their parental WT forms. Wild type variants of S. epidermidis phosphorylated Akt at similar or lower levels as control uninfected cells. Next, the induction of mTOR target, phosphorylated ribosomal protein S6, was measured in bacteria-infected macrophages. The level of phosphorylation was significantly reduced when the cells were exposed to WT strains of S. epidermidis. In contrast, the SCV strains activated S6 protein mostly at a level comparable to the control cells. Rapamycin inhibited mTOR activation as the number of p-S6 positive cells decreased in the tested cases. To conclude, the SCV strains activate the PI3K-Akt signaling pathway in opposite to WT strains. This fact however did not influence the increase in the number of live SCV bacteria as compared to the WT strains. Knowing that the PI3K-Akt pathway is involved in proinflammatory cytokines suppression, SCVs seem to use this pathway to reduce the inflammatory response during the infection.

[Mechanisms of myeloid cell RelA/p65 in cigarette smoking-induced lung cancer growth in mice].

PubMed

Yao, Yiwen; Wu, Junlu; Quan, Wenqiang; Zhou, Hong; Zhang, Yu; Wan, Haiying; Li, Dong

2014-06-01

The aim of this study was to investigate the mechanism of cigarette smoking (CS)-induced lung cancer growth in mice. RelA/p65⁻/⁻ mice and WT mice were used to establish mouse models of lung cancer. Both mice were divided into two groups: air group and CS group, respectively. Tumor number on the lung surface was counted and maximal tumor size was evaluated using HE staining. Kaplan Meier (K-M) survival curve was used to analyze the survival rate of the mice. Expression of Ki-67, TNF-α and CD68 in the tumor tissue was determined by immunohistochemical analysis, and cyclin D1 and c-myc proteins were examined by Western blot. Apoptosis of tumor cells was analyzed using TUNEL staining. The concentrations of inflammatory cytokines TNF-α, IL-6 and KC in the mouse lung tissues were evaluated by ELISA. Compared with the WT air group, the lung weight, lung tumor multiplicity, as well as maximum tumor size in the WT mice exposed to CS were (1.5 ± 0.1)g, (64.8 ± 4.1) and (7.6 ± 0.2) mm, respectively, significantly increased than those in the WT mice not exposed to CS (P < 0.05 for all). However, there were no statistically significant differences between RelA/p65⁻/⁻ mice before and after CS exposure (P > 0.05 for all). Kaplan-Meier survival analysis showed that CS exposure significantly shortened the life time of WT mice (P < 0.05), and deletion of RelA/p65 in myeloid cells resulted in an increased survival compared with that of the WT mice (P < 0.05 for all). The ratios of Ki-67 positive tumor cells were (43.4 ± 2.9)%, (60.6 ± 5.4)%, (12.8 ± 3.6)% and (15.0 ± 4.2)% in the WT air group, WT CS groups, RelA/p65⁻/⁻ air groups and RelA/p65⁻/⁻ CS groups, respectively. After smoking, the number of Ki-67-positive cells was significantly increased in the WT mice (P < 0.05). However, there was no significant difference between the RelA/p65⁻/⁻ groups before and after smoking (P > 0.05). The apoptosis rate of WT air, WT CS, RelA/p65⁻/⁻ air and RelA/p65⁻/⁻ CS groups were (11.6 ± 1.7)%, (13.0 ± 2.0)%, (13.2 ± 2.0)% and (11.0 ± 1.4)%, respectively, with no significant difference among them (P > 0.05). Expression of cyclin D1 and c-myc was induced in response to CS exposure in lung tumor cells of WT mice. In contrast, their expressions were not significantly changed in the RelA/p65⁻/⁻ mice after smoke exposure. CS exposure was associated with an increased number of macrophages infiltrating in the tumor tissue, in both WT and RelA/p65⁻/⁻ mice (P < 0.05). The concentrations of IL-6, KC and TNF-α were significantly increased after CS exposure in the lungs of WT mice (P < 0.05). Cigarette smoking promotes the lung cancer growth in mice. Myeloid cell RelA/p65 mediates CS-induced tumor growth. TNFα regulated by RelA/p65 may be involved in the lung cancer development.

Characterization and human gingival fibroblasts biocompatibility of hydroxyapatite/PMMA nanocomposites for provisional dental implant restoration

NASA Astrophysics Data System (ADS)

Zhang, Jingchao; Liao, Juan; Mo, Anchun; Li, Yubao; Li, Jidong; Wang, Xuejiang

2008-11-01

The aim of this study was to determine nHA/PMMA composites (H/P) in an optimal ratio with improved cytocompatibility as well as valid physical properties for provisional dental implant restoration. 20 wt.%, 30 wt.%, 40 wt.% and 50 wt.% H/P were developed and characterized using XPS, bending strength test and SEM. Human gingival fibroblasts cultured in extracts or directly on sample discs were investigated by fluorescent staining and MTT assay. Chemical integration in nHA/PMMA interface was indicated by XPS. Typical fusiform cells with adhesion spots were detected on H/P discs. MTT results also indicated higher cell viability in 30 wt.% and 40 wt.% H/P discs ( P < 0.05). We conclude that nHA addition to PMMA enhances cytocompatibility and the optimal nHA/PMMA ratio for provisional fixed crowns (PFC) is 0.4:1.

A study of electrochemical devices based on Agar-Agar-NH4I biopolymer electrolytes

NASA Astrophysics Data System (ADS)

Selvalakshmi, S.; Mathavan, T.; Selvasekarapandian, S.; Premalatha, M.

2018-04-01

A polymer electrolyte system has been developed using a biopolymer namely, Agar-Agar in combination with ammonium iodide in different weight percentages by solution casting technique. The films were characterized electrically by AC Impedance Spectroscopy for its conductivity. The highest conductivity achieved at room temperature was for 50 wt. % agar-agar: 50 wt. % NH4I with a conductivity value of 1.20 × 10-4 Scm-1. An electrochemical cell was fabricated in the configuration of: Zn + ZnSO4.7H2O + graphite (anode) | 50 wt. % (Agar-agar): 50 wt. % NH4I (electrolyte) | PbO2 + V2O5 + graphite (cathode) and it produced a maximum open circuit voltage of 1.73 V. A single PEM fuel cell was constructed with the highest conducting sample (50 wt. % (Agar-agar): 50 wt. % NH4I) and it exhibited an output voltage of 408mV.

Modulation of growth, differentiation, and mucous glycoprotein synthesis by retinyl acetate in cloned carcinoma cell lines. [Rats

DOE Office of Scientific and Technical Information (OSTI.GOV)

Marchok, A.C.; Clark, J.N.; Klein-Szanto, A.

1981-06-01

The ability of retinyl acetate to alter growth, differentiation, and synthesis of mucous glycoproteins in cell lines cloned from an adenocarcinoma (T-8) and a squamous cell carcinoma (1000 WT) was investigated with the use of F344 rats. Growth rate was inhibited approximately 25 and 50% in 6.6 x 10/sup -6/ and 3.3 x 10/sup -5/ M retinyl acetate, respectively, in both cell lines. Retinyl acetate induced the formation of numerous vacuoles and periodic acid-silver methenamine-positive granules in both T-8 and 1000 WT cells. After T-8 cells were cultured for 7 days in retinyl acetate, (/sup 3/H)glucosamine incorporation increased 133- tomore » 147-fold and (/sup 14/C)serine incorporation increased twelvefold to twentyfold in the high-molecular-weight mucous glycoprotein fraction (peak A) from the cell cytosol. In 1000 WT cells, (/sup 3/H)glucosamine incorporation increased only 4.2- to 7.5-fold, and (/sup 14/C)serine incorporation increased only 2.6- to 4.6-fold under the same culture conditions. Thus T-8 cells showed a marked increase in the synthesis and secretion of mucins, whereas 1000 WT cells showed a comparatively small but significant increase.« less

Amish burn ointment and burdock leaf dressings: assessments of antimicrobial and cytotoxic activities.

PubMed

Rieman, Mary T; Neely, Alice N; Boyce, Steven T; Kossenjans, William J; Durkee, Paula J; Zembrodt, Jacquelyn M; Puthoff, Barbara K; Kagan, Richard J

2014-01-01

Amish burn wound ointment (ABO) contains honey, lanolin, oils, glycerin, bees wax, and other natural additives. Although there are many anecdotal reports that this ointment covered with a burdock leaf (BL) dressing promotes burn wound healing, little scientific testing of this treatment has occurred. The goal of this study was to evaluate in vitro some of the components of this treatment modality for antimicrobial and cytotoxic activities. The ABO was tested for sterility using standard microbiological techniques. Because of the semisolid, lipid-based nature of the salve, the at-use product could not be tested in bioassays. Samples of BL and the dry ingredients (DI) used in the ointment were provided by the Amish vendor. Aqueous extracts of the DI and of the BL were prepared and freeze dried. The freeze-dried extracts were reconstituted, filtered, and tested separately on keratinocyte and fibroblast cell cultures for cytotoxicity (growth inhibition assay) and against a panel of susceptible and resistant microbes for antimicrobial activity (Nathan's agar-well diffusion assay) in a series of concentrations (% wt/vol). Neither DI nor BL extracts demonstrated antimicrobial activity against any of organisms tested. The DI extract inhibited growth of both keratinocytes and fibroblasts at the 0.1% concentration. The 0.1 and 0.03% concentrations of the BL extract were cytotoxic to both keratinocytes and fibroblasts. Although tests for microbial growth from the at-use preparation of the ABO were negative, extracts of the DI and BL did not demonstrate any antimicrobial activity. Additionally, both extracts inhibited the growth of skin cells in vitro at higher concentrations. These results suggest caution in the use of ABO and BL dressings if there is more than a minimal risk of complications from the burn injury.

[Study of negative feedback between wild-type BRAF or RAFV600E and Mps1 in melanoma].

PubMed

Zhang, Ling; He, Chanting; Bi, Yanghui; Liu, Feng; Cui, Heyang; Wang, Juan; Song, Bin; Shi, Ruyi; Yang, Bin; Wang, Fang; Jia, Zhiwu; Zhao, Zhenxiang; Liu, Jing

2015-04-01

To study the effect of Mps1 on BRAFWT/MEK/ERK pathway in the presence of wild type BRAF or BRAFV600E in melanoma. Melanoma cells harboring BRAFWT genotype were transfected either with pBabe-puro-GST-BRAF-WT and/or pBabe-puro-GFP-Mps1-WT or pBabe-puro-GST-BRAFV600E and/or pBabe-puro-GFP-Mps1-WT, followed by Western blot to detect Mps1 and p-ERK expression. The melanoma cells harboring BRAFWT and BRAFV600E genotype were infected with pSUPER-Mps1 retrovirus to knockdown the endogenous Mps1 protein, followed by Western blot to detect Mps1 and p-ERK expression. Meanwhile, melanoma cells harboring BRAFV600E genotype were infected with pBabe-puro-GFP-Mps1 and Western blot was performed to detect Mps1 and p-ERK expression. In melanoma cells harboring BRAFWT genotype and transfected with pBabe-puro-GST-BRAF-WT and pBabe-puro-GFP-Mps1-WT, phospho-ERK levels were notably reduced as compared to either negative control or empty vector. However, cells transfected with pBabe-puro-GST-BRAFV600E and pBabe-puro-GFP-Mps1-WT, phospho-ERK levels did not change significantly compared with either negative control or empty vector. Knockout of Mps1 in BRAF wild-type cell lines led to an increased ERK activity. However, there was no significant change of ERK activity in BRAFV600E cell lines in the absence of Mps1. The expression of p-ERK in BRAFV600E mutant cell lines infected with pBabe-puro-GFP-Mps1-WT did not show any significant difference from either negative control or empty vector. Based on these findings, it suggests that there exists an auto-regulatory negative feedback loop between the Mps1 kinase and BRAFWT/ERK signaling. Oncogenic BRAFV600E abrogates the regulatory negative feedback loop of Mps1 on the MAPK pathway.

Graphene oxide nanoflakes incorporated gelatin-hydroxyapatite scaffolds enhance osteogenic differentiation of human mesenchymal stem cells

NASA Astrophysics Data System (ADS)

Nair, Manitha; Nancy, D.; Krishnan, Amit G.; Anjusree, G. S.; Vadukumpully, Sajini; Nair, Shantikumar V.

2015-04-01

In this study, graphene oxide (GO) nanoflakes (0.5 and 1 wt%) were incorporated into a gelatin-hydroxyapatite (GHA) matrix through a freeze drying technique and its effect to enhance mechanical strength and osteogenic differentiation was studied. The GHA matrix with GO demonstrated less brittleness in comparison to GHA scaffolds. There was no significant difference in mechanical strength between GOGHA0.5 and GOGHA1.0 scaffolds. When the scaffolds were immersed in phosphate buffered saline (to mimic physiologic condition) for 60 days, around 50-60% of GO was released in sustained and linear manner and the concentration was within the toxicity limit as reported earlier. Further, GOGHA0.5 scaffolds were continued for cell culture experiments, wherein the scaffold induced osteogenic differentiation of human adipose derived mesenchymal stem cells without providing supplements like dexamethasone, L-ascorbic acid and β glycerophosphate in the medium. The level of osteogenic differentiation of stem cells was comparable to those cultured on GHA scaffolds with osteogenic supplements. Thus biocompatible, biodegradable and porous GO reinforced gelatin-HA 3D scaffolds may serve as a suitable candidate in promoting bone regeneration in orthopaedics.

Global gene expression analysis of transgenic, mannitol-producing, and salt-tolerant Arabidopsis thaliana indicates widespread changes in abiotic and biotic stress-related genes

PubMed Central

Chan, Zhulong; Grumet, Rebecca; Loescher, Wayne

2011-01-01

Mannitol is a putative osmoprotectant contributing to salt tolerance in several species. Arabidopsis plants transformed with the mannose-6-phosphate reductase (M6PR) gene from celery were dramatically more salt tolerant (at 100 mM NaCl) as exhibited by reduced salt injury, less inhibition of vegetative growth, and increased seed production relative to the wild type (WT). When treated with 200 mM NaCl, transformants produced no seeds, but did bolt, and exhibited less chlorosis/necrosis and greater survival and dry weights than the WT. Without salt there were no M6PR effects on growth or phenotype, but expression levels of 2272 genes were altered. Many fewer differences (1039) were observed between M6PR and WT plants in the presence of salt, suggesting that M6PR pre-conditioned the plants to stress. Previous work suggested that mannitol is an osmoprotectant, but mannitol levels are invariably quite low, perhaps inadequate for osmoprotectant effects. In this study, transcriptome analysis reveals that the M6PR transgene activated the downstream abscisic acid (ABA) pathway by up-regulation of ABA receptor genes (PYL4, PYL5, and PYL6) and down-regulation of protein phosphatase 2C genes (ABI1 and ABI2). In the M6PR transgenic lines there were also increases in transcripts related to redox and cell wall-strengthening pathways. These data indicate that mannitol-enhanced stress tolerance is due at least in part to increased expression of a variety of stress-inducible genes. PMID:21821598

BRE facilitates skeletal muscle regeneration by promoting satellite cell motility and differentiation

PubMed Central

Xiao, Lihai; Lee, Kenneth Ka Ho

2016-01-01

ABSTRACT The function of the Bre gene in satellite cells was investigated during skeletal muscle regeneration. The tibialis anterior leg muscle was experimentally injured in Bre knockout mutant (BRE-KO) mice. It was established that the accompanying muscle regeneration was impaired as compared with their normal wild-type counterparts (BRE-WT). There were significantly fewer pax7+ satellite cells and smaller newly formed myofibers present in the injury sites of BRE-KO mice. Bre was required for satellite cell fusion and myofiber formation. The cell fusion index and average length of newly-formed BRE-KO myofibers were found to be significantly reduced as compared with BRE-WT myofibers. It is well established that satellite cells are highly invasive which confers on them the homing ability to reach the muscle injury sites. Hence, we tracked the migratory behavior of these cells using time-lapse microscopy. Image analysis revealed no difference in directionality of movement between BRE-KO and BRE-WT satellite cells but there was a significant decrease in the velocity of BRE-KO cell movement. Moreover, chemotactic migration assays indicated that BRE-KO satellite cells were significantly less responsive to chemoattractant SDF-1α than BRE-WT satellite cells. We also established that BRE normally protects CXCR4 from SDF-1α-induced degradation. In sum, BRE facilitates skeletal muscle regeneration by enhancing satellite cell motility, homing and fusion. PMID:26740569

Mercury demethylation in waterbird livers: Dose-response thresholds and differences among species

USGS Publications Warehouse

Eagles-Smith, Collin A.; Ackerman, Joshua T.; Julie, Y.E.E.; Adelsbach, T.L.

2009-01-01

We assessed methylmercury (MeHg) demethylation in the livers of adults and chicks of four waterbird species that commonly breed in San Francisco Bay: American avocets, black-necked stilts, Caspian terns, and Forster's terns. In adults (all species combined), we found strong evidence for a threshold, model where MeHg demethylation occurred above a hepatic total mercury concentration threshold of 8.51 ?? 0.93 ??g/g dry weight, and there was a strong decline in %MeHg values as total mercury (THg) concentrations increased above 8.51 ??g/g dry weight. Conversely, there was no evidence for a demethylation threshold in chicks, and we found that %MeHg values declined linearly with increasing THg concentrations. For adults, we also found taxonomie differences in the demethylation responses, with avocets and stilts showing a higher demethylation rate than that of terns when concentrations exceeded the threshold, whereas terns had a lower demethylation threshold (7.48 ?? 1.48 ??g/g dry wt) than that of avocets and stilts (9.91 ?? 1.29 ??g/g dry wt). Finally, we assessed the role of selenium (Se) in the demethylation process. Selenium concentrations were positively correlated with inorganic Hg in livers of birds above the demethylation threshold but not below. This suggests that Se may act as a binding site for demethylated Hg and may reduce the potential for secondary toxicity. Our findings indicate that waterbirds demethylate mercury in their livers if exposure exceeds a threshold value and suggest that taxonomie differences in demethylation ability may be an important factor in evaluating species-specific risk to MeHg exposure. Further, we provide strong evidence for a threshold of approximately 8.5 ??g/g dry weight of THg in the liver where demethylation is initiated. ?? 2009 SETAC.

Extraction of caustic potash from spent tea for biodiesel Production

NASA Astrophysics Data System (ADS)

Sulaiman, Sarina; Faiz Che Fisol, Ahmad; Sharikh, Atikah Mohamed; Noraini Jimat, Dzun; Jamal, Parveen

2018-01-01

Biodiesel is an alternative to non-renewable fossil fuels due to its low gas emission and economical value. This study aims to extract caustic potash (KOH) from spent tea and to optimize the transesterfication process based on parameters such as amount of catalyst, reaction temperature and methanol to oil ratio. The spent tea was first dried at 60°C prior to calcination at 600°C for two hours. Caustic Potash were extracted from the calcined spent tea. The transesterification process was done based on Design of Experiments (DOE) to study the effects of amount of catalyst ranging from 0.5 wt % to 2.5 wt %, reaction temperature from 55°C to 65°C and methanol to oil ratio from 6:1 to 12:1 at a constant agitation rate of 300 rpm for three hours. The calcined spent tea produced was recorded the highest at 54.3 wt % and the extracted catalyst was 2.4 wt %. The optimized biodiesel yield recorded was 56.95% at the optimal conditions of 2.5 wt % amount of catalyst, 65°C reaction temperature and 9:1 methanol to oil ratio.

Two distinct HLA-A0201-presented epitopes of the Wilms tumor antigen 1 can function as targets for leukemia-reactive CTL.

PubMed

Bellantuono, Ilaria; Gao, Liquan; Parry, Suzanne; Marley, Steve; Dazzi, Francesco; Apperley, Jane; Goldman, John M; Stauss, Hans J

2002-11-15

Using the allo-restricted T-cell approach to circumvent tolerance, we have previously identified a cytotoxic T-lymphocyte (CTL) epitope in the transcription factor Wilms tumor antigen 1 (WT1) presented by HLA-A0201 (A2) class I molecules. Here we describe an additional A2-presented epitope and show that CTLs against both epitopes kill WT1-expressing leukemia cell lines. Colony-forming assays demonstrated that both types of CTL killed CD34(+) progenitor cells from A2(+) leukemia patients, but not from A2(+) healthy individuals. The long-term culture-initiating cell (LTC-IC) assay was used to analyze the killing activity of WT1-specific CTLs against the more immature fraction of CD34(+) cells. The CTLs killed LTC-ICs of patients with chronic myelogenous leukemia (CML), whereas the function of normal CD34(+) progenitor/stem cells was not inhibited. Together, the data show that CTLs specific for 2 distinct peptide epitopes of WT1 can discriminate between normal and leukemia LTC-ICs, suggesting that such CTLs have the potential to selectively kill CML progenitor/stem cells.

Filamin A Mediates Wound Closure by Promoting Elastic Deformation and Maintenance of Tension in the Collagen Matrix

PubMed Central

Mohammadi, Hamid; Pinto, Vanessa I.; Wang, Yongqiang; Hinz, Boris; Janmey, Paul A.; McCulloch, Christopher A.

2016-01-01

Cell-mediated remodeling and wound closure are critical for efficient wound healing, but the contribution of actin-binding proteins to contraction of the extracellular matrix is not defined. We examined the role of filamin A (FLNa), an actin filament cross-linking protein, in wound contraction and maintenance of matrix tension. Conditional deletion of FLNa in fibroblasts in mice was associated with ~ 4 day delay of full-thickness skin wound contraction compared with wild-type (WT) mice. We modeled the healing wound matrix using cultured fibroblasts plated on grid-supported collagen gels that create lateral boundaries, which are analogues to wound margins. In contrast to WT cells, FLNa knockdown (KD) cells could not completely maintain tension when matrix compaction was resisted by boundaries, which manifested as relaxed matrix tension. Similarly, WT cells on cross-linked collagen, which requires higher levels of sustained tension, exhibited approximately fivefold larger deformation fields and approximately twofold greater fiber alignment compared with FLNa KD cells. Maintenance of boundary-resisted tension markedly influenced the elongation of cell extensions: in WT cells, the number (~50%) and length (~300%) of cell extensions were greater than FLNa KD cells. We conclude that FLNa is required for wound contraction, in part by enabling elastic deformation and maintenance of tension in the matrix. PMID:26134946

Activation of Dll4/Notch Signaling and Hypoxia-Inducible Factor-1 Alpha Facilitates Lymphangiogenesis in Lacrimal Glands in Dry Eye.

PubMed

Min, Ji Hwan; Lee, Chul Hee; Ji, Yong Woo; Yeo, Areum; Noh, Hyemi; Song, Insil; Kim, Eung Kweon; Lee, Hyung Keun

2016-01-01

By using hypoxia-inducible factor-1 alpha conditional knockout (HIF-1α CKO) mice and a dry eye (DE) mouse model, we aimed to determine the role played by delta-like ligand 4 (Dll4)/Notch signaling and HIF-1α in the lymphangiogenesis of lacrimal glands (LGs). C57BL/6 mice were housed in a controlled-environment chamber for DE induction. During DE induction, the expression level of Dll4/Notch signaling and lymphangiogenesis in LGs was measured by quantitative RT-PCR, immunoblot, and immunofluorescence staining. Next, lymphangiogenesis was measured after Dll4/Notch signal inhibition by anti-Dll4 antibody or γ-secretase inhibitor. Using HIF-1α CKO mice, the expression of Dll4/Notch signaling and lymphangiogenesis in LGs of DE-induced HIF-1α CKO mice were assessed. Additionally, the infiltration of CD45+ cells in LGs was assessed by immunohistochemical (IHC) staining and flow cytometry for each condition. DE significantly upregulated Dll4/Notch and lymphangiogenesis in LGs. Inhibition of Dll4/Notch significantly suppressed lymphangiogenesis in LGs. Compared to wild-type (WT) mice, DE induced HIF-1α CKO mice showed markedly low levels of Dll4/Notch and lymphangiogenesis. Inhibition of lymphangiogenesis by Dll4/Notch suppression resulted in increased CD45+ cell infiltration in LGs. Likewise, CD45+ cells infiltrated more in the LGs of HIF-1α CKO DE mice than in non-DE HIF-1α CKO mice. Dll4/Notch signaling and HIF-1α are closely related to lymphangiogenesis in DE-induced LGs. Lymphangiogenesis stimulated by Dll4/Notch and HIF-1α may play a role in protecting LGs from DE-induced inflammation by aiding the clearance of immune cells from LGs.

Investigation on Sr0.2Na0.8Nb1-xVxO3 (x=0.1, 0.2, 0.3) as new ceramic anode materials for low-temperature solid oxide fuel cells

NASA Astrophysics Data System (ADS)

Pan, Ke-Ji; Hussain, A. Mohammed; Wachsman, Eric D.

2017-04-01

Variants of SNNV (Sr0.2Na0.8Nb1-xVxO3, X = 0.1-0.3) ceramic oxides were synthesized via wet chemical method. SNNVs show high electronic conductivity of >100 S/cm when reduced in hydrogen at a relatively low temperature of 650 °C. In particular, 30% V-doped SNNV exhibited the highest conductivity of 300 S/cm at 450 °C. In order to investigate the fuel cell performance, Gd0.1Ce0.9O2-δ (GDC) based electrolyte-supported fuel cells were prepared to study the anode characteristics. Sr0.2Na0.8Nb0.9V0.1O3 (SNNV10)-GDC composite was used as an anode and La0.6Sr0.4Co0.2Fe0.8O3-δ (LSCF)-GDC as a cathode. Both electrodes were porous and sintered at 1050 °C for 2 h in air. The anode side of the fuel cell was infiltrated with 10 wt% GDC/Ni-GDC precursor to activate the anode for fuel oxidation. I-V characteristics were determined in gas conditions such as dry/humidified hydrogen and methane at 650 °C. With the infiltration Ni-GDC, peak power density (PPD) of 280 mW/cm2 and 220 mW/cm2 in dry H2 and CH4, respectively, were obtained at 650 °C, which is higher than GDC alone as infiltrate. The high resistances in the humidified conditions are attributed to the lower conductivity of SNNV10 in high PO2 atmospheres.

Identification of a Novel C-Terminal Truncated WT1 Isoform with Antagonistic Effects against Major WT1 Isoforms

PubMed Central

Tatsumi, Naoya; Hojo, Nozomi; Sakamoto, Hiroyuki; Inaba, Rena; Moriguchi, Nahoko; Matsuno, Keiko; Fukuda, Mari; Matsumura, Akihide; Hayashi, Seiji; Morimoto, Soyoko; Nakata, Jun; Fujiki, Fumihiro; Nishida, Sumiyuki; Nakajima, Hiroko; Tsuboi, Akihiro; Oka, Yoshihiro; Hosen, Naoki; Sugiyama, Haruo; Oji, Yusuke

2015-01-01

The Wilms’ tumor gene WT1 consists of 10 exons and encodes a zinc finger transcription factor. There are four major WT1 isoforms resulting from alternative splicing at two sites, exon 5 (17AA) and exon 9 (KTS). All major WT1 isoforms are overexpressed in leukemia and solid tumors and play oncogenic roles such as inhibition of apoptosis, and promotion of cell proliferation, migration and invasion. In the present study, a novel alternatively spliced WT1 isoform that had an extended exon 4 (designated as exon 4a) with an additional 153 bp (designated as 4a sequence) at the 3’ end was identified and designated as an Ex4a(+)WT1 isoform. The insertion of exon 4a resulted in the introduction of premature translational stop codons in the reading frame in exon 4a and production of C-terminal truncated WT1 proteins lacking zinc finger DNA-binding domain. Overexpression of the truncated Ex4a(+)WT1 isoform inhibited the major WT1-mediated transcriptional activation of anti-apoptotic Bcl-xL gene promoter and induced mitochondrial damage and apoptosis. Conversely, suppression of the Ex4a(+)WT1 isoform by Ex4a-specific siRNA attenuated apoptosis. These results indicated that the Ex4a(+)WT1 isoform exerted dominant negative effects on anti-apoptotic function of major WT1 isoforms. Ex4a(+)WT1 isoform was endogenously expressed as a minor isoform in myeloid leukemia and solid tumor cells and increased regardless of decrease in major WT1 isoforms during apoptosis, suggesting the dominant negative effects on anti-apoptotic function of major WT1 isoforms. These results indicated that Ex4a(+)WT1 isoform had an important physiological function that regulated oncogenic function of major WT1 isoforms. PMID:26090994

New Nitric Oxide Donor NCX 1443: Therapeutic Effects on Pulmonary Hypertension in the SAD Mouse Model of Sickle Cell Disease.

PubMed

Abid, Shariq; Kebe, Kanny; Houssaïni, Amal; Tomberli, Françoise; Marcos, Elisabeth; Bizard, Emilie; Breau, Marielle; Parpaleix, Aurelien; Tissot, Claire-Marie; Maitre, Bernard; Lipskaia, Larissa; Derumeaux, Genevieve; Bastia, Elena; Mekontso-Dessap, Armand; Adnot, Serge

2018-05-01

Nitric oxide (NO) donors may be useful for treating pulmonary hypertension (PH) complicating sickle cell disease (SCD), as endogenous NO is inactivated by hemoglobin released by intravascular hemolysis. Here, we investigated the effects of the new NO donor NCX1443 on PH in transgenic SAD mice, which exhibit mild SCD without severe hemolytic anemia. In SAD and wild-type (WT) mice, the pulmonary pressure response to acute hypoxia was similar and was abolished by 100 mg/kg NCX1443. The level of PH was also similar in SAD and WT mice exposed to chronic hypoxia (9% O2) alone or with SU5416 and was similarly reduced by daily NCX1443 gavage. Compared with WT mice, SAD mice exhibited higher levels of HO-1, endothelial NO synthase, and PDE5 but similar levels of lung cyclic guanosine monophosphate. Cultured pulmonary artery smooth muscle cells from SAD mice grew faster than those from WT mice and had higher PDE5 protein levels. Combining NCX1443 and a PDE5 inhibitor suppressed the growth rate difference between SAD and WT cells and induced a larger reduction in hypoxic PH severity in SAD than in WT mice. By amplifying endogenous protective mechanisms, NCX1443 in combination with PDE5 inhibition may prove useful for treating PH complicating SCD.

Expression of Biologically Active Human Butyrylcholinesterase in the Cabbage Looper (Trichoplusia ni)

DTIC Science & Technology

2000-01-01

recombinant human BUChE; Sf, Spodoptera frugiperda ; VX, 0-ethyl S-[2-[bis(I -methylethyl)amino]ethyl]methyl phosphonothiolate; wt, wild-type. 1 To whom...ATCC (Rockville, MD, U.S.A.). Insect cells ( Spodoptera frugiperda Sf9 cells and T. ni High 5 cells) and wild-type (wt)-AcNPV were purchased from

Loss of Myeloid Related Protein-8/14 Exacerbates Cardiac Allograft Rejection

PubMed Central

Shimizu, Koichi; Libby, Peter; Rocha, Viviane Z.; Folco, Eduardo J.; Shubiki, Rica; Grabie, Nir; Jang, Sunyoung; Lichtman, Andrew H.; Shimizu, Ayako; Hogg, Nancy; Simon, Daniel I.; Mitchell, Richard N.; Croce, Kevin

2011-01-01

Background The calcium-binding proteins myeloid-related protein (MRP)-8 (S100A8) and MRP-14 (S100A9) form MRP-8/14 heterodimers (S100A8/A9, calprotectin) that regulate myeloid cell function and inflammatory responses, and serve as early serum markers for monitoring acute allograft rejection. Despite functioning as a pro-inflammatory mediator, the pathophysiological role of MRP-8/14 complexes in cardiovascular disease is incompletely defined. This study investigated the role of MRP-8/14 in cardiac allograft rejection using MRP-14-deficient mice (MRP14-/-) that lack MRP-8/14 complexes. Methods and Results We examined parenchymal rejection (PR) after major histocompatibility complex (MHC) class II allomismatched cardiac transplantation (bm12 donor heart and B6 recipients) in wild-type (WT) and MRP14-/- recipients. Allograft survival averaged 5.9 ± 2.9 weeks (n=10) in MRP14-/- recipients, compared to > 12 weeks (n = 15, p < 0.0001) in WT recipients. Two weeks after transplantation, allografts in MRP14-/- recipients had significantly higher PR scores (2.8 ± 0.8, n=8) than did WT recipients (0.8 ± 0.8, n=12, p<0.0001). Compared to WT recipients, allografts in MRP14-/- recipients had significantly increased T-cell and macrophage infiltration, as well as increased mRNA levels of IFN-γ and IFN-γ–associated chemokines (CXCL9, CXCL10, and CXCL11), IL-6, and IL-17, with significantly higher levels of Th17 cells. MRP14-/- recipients also had significantly more lymphocytes in the adjacent paraaortic lymph nodes than did WT recipients (cell number per lymph node: 23.7 ± 0.7 × 105 for MRP14-/- vs. 6.0 ± 0.2 × 105 for WT, p < 0.0001). The dendritic cells (DCs) of the MRP14-/- recipients of bm12 hearts expressed significantly higher levels of the co-stimulatory molecules CD80 and CD86 than did those of WT recipients 2 weeks after transplantation. Mixed leukocyte reactions using allo-EC-primed MRP14-/- DCs resulted in significantly higher antigen-presenting function than reactions using WT DCs. Ovalbumin-primed MRP14-/- DCs augmented proliferation of OT-II CD4+ T cells with increased IL-2 and IFN-γ production. Cardiac allografts of B6 MHC class II-/- hosts and of B6 WT hosts receiving MRP14-/- DCs had significantly augmented inflammatory cell infiltration and accelerated allograft rejection, compared to WT DCs from transferred recipient allografts. Bone marrow–derived MRP14-/- DCs infected with MRP-8 and MRP-14 retroviral vectors showed significantly decreased CD80 and CD86 expression compared to controls, indicating that MRP-8/14 regulates B7-costimulatory molecule expression. Conclusion Our results indicate that MRP-14 regulates B7 molecule expression and reduces antigen presentation by DCs, and subsequent T-cell priming. The absence of MRP-14 markedly increased T-cell activation and exacerbated allograft rejection, indicating a previously unrecognized role for MRP-14 in immune cell biology. PMID:22144572

Bach2 Controls Homeostasis of Eosinophils by Restricting the Type-2 Helper Function of T Cells.

PubMed

Sato, Yuki; Kato, Hiroki; Ebina-Shibuya, Risa; Itoh-Nakadai, Ari; Okuyama, Ryuhei; Igarashi, Kazuhiko

2017-03-01

Bach2 is a transcription factor which represses its target genes and plays important roles in the differentiation of B and T lymphoid cells. Bach2-deficient (KO) mice develop severe pulmonary alveolar proteinosis, which is associated with increased numbers of granulocytes and T cells. Bach2 is essential for the regulation of T cells, but its role in the regulation of granulocytes is not clear. Here, we observed increased numbers of eosinophils but not neutrophils in the bone marrow, spleen, peripheral blood, and bronchoalveolar lavage fluids of Bach2 KO mice compared with those of wild-type (WT) mice. Upon co-transplantation of the bone marrow cells from CD45.2 Bach2 KO and CD45.1/CD45.2 double-positive WT mice to irradiated WT CD45.1/CD45.2 mice, the reconstituted numbers of eosinophils were similar between Bach2 KO and WT cells. These results showed that the deficiency of Bach2 in eosinophils did not directly drive the differentiation of eosinophils. To investigate the effect of Bach2 KO CD4 + T cells upon eosinophils, we analyzed Rag2/Bach2-double deficient (dKO) mice which lack lymphocytes including CD4 + T cells. Rag2/Bach2 dKO mice did not show any increase in the numbers of eosinophils. Importantly, Bach2 KO mice showed an increase of interleukin-5 (Il-5) in the sera compared with WT mice. These results suggest that up-regulated functions of CD4 + T cells including secretion of Il-5 resulted in proliferation and/or migration to peripheral tissues of eosinophils in Bach2 KO mice. We propose that Bach2 controls homeostasis of eosinophils via restricting the production of Il-5 in CD4 + T cells.

Chemically characterized Mentha cardiaca L. essential oil as plant based preservative in view of efficacy against biodeteriorating fungi of dry fruits, aflatoxin secretion, lipid peroxidation and safety profile assessment.

PubMed

Dwivedy, Abhishek Kumar; Prakash, Bhanu; Chanotiya, Chandan Singh; Bisht, Deepa; Dubey, Nawal Kishore

2017-08-01

The study reports Mentha cardiaca essential oil (EO) as plant based preservative against fungal and aflatoxin contamination of stored dry fruits. Mycoflora analysis of the dry fruits revealed Aspergillus favus LHP-PV-1 as the most aflatoxigenic isolate with highest Aflatoxin B 1 content. M. cardiaca EO showed broad fungitoxic spectrum inhibiting the tested moulds contaminating dry fruits. It's minimum inhibitory concentration (MIC), minimum aflatoxin inhibitory concentration (MAIC) and minimum fungicidal concentration (MFC) against A. favus LHP-PV-1 were recorded to be 1.25, 1.0 and 2.25 µL/mL respectively. The EO caused decrease in ergosterol content and enhanced leakage of Ca 2+ , K + and Mg 2+ ions from treated fungal cells, depicting fungal plasma membrane as the site of antifungal action. The EO showed promising DPPH free radical scavenging activity (IC 50 value:15.89 µL/mL) and favourable safety profile with LD 50 value (7133.70 mg/kg body wt.) when estimated through acute oral toxicity on mice. Carvone (61.62%) was recorded as the major component of the oil during chemical characterisation through GC-MS. Based on strong antifungal, antiaflatoxigenic and antioxidant potential, the chemically characterised M. cardiaca EO may be recommended as safe plant based preservative and shelf life enhancer of food items. This is the first report on antifungal and antiaflatoxigenic activity of M. cardiaca EO. Copyright © 2017 Elsevier Ltd. All rights reserved.

Differential effects of Phe19 and Phe20 on fibril formation by amyloidogenic peptide A beta 16-22 (Ac-KLVFFAE-NH2).

PubMed

Inouye, Hideyo; Gleason, Katherine A; Zhang, Dong; Decatur, Sean M; Kirschner, Daniel A

2010-08-01

The sequence KLVFFAE (A beta 16-22) in Alzheimer's beta-amyloid is thought to be a core beta-structure that could act as a template for folding other parts of the polypeptide or molecules into fibrillar assemblies rich in beta-sheet. To elucidate the mechanism of the initial folding process, we undertook combined X-ray fiber/powder diffraction and infrared (IR) spectroscopy to analyze lyophilized A beta 16-22 and solubilized/dried peptide containing nitrile probes at F19 and/or F20. Solubilized/dried wild-type (WT) A beta 16-22 and the peptide containing cyanophenylalanine at F19 (19CN) or at F20 (20CN) gave fiber patterns consistent with slab-like beta-crystallites that were cylindrically averaged around the axis parallel to the polypeptide chain direction. The WT and 19CN assemblies showed 30-A period arrays arising from the stacking of the slabs along the peptide chain direction, whereas the 20CN assemblies lacked any such stacking. The electron density projection along the peptide chain direction indicated similar side-chain dispositions for WT and 20CN, but not for 19CN. These X-ray results and modeling imply that in the assembly of WT A beta 16-22 the F19 side chain is localized within the intersheet space and is involved in hydrophobic contact with amino acids across the intersheet space, whereas the F20 side chain localized near the slab surface is less important for the intersheet interaction, but involved in slab stacking. IR observations for the same peptides in dilute solution showed a greater degree of hydrogen bonding for the nitrile groups in 20CN than in 19CN, supporting this interpretation. (c) 2010 Wiley-Liss, Inc.

The pro-apoptotic protein Bim is a convergence point for cAMP/protein kinase A- and glucocorticoid-promoted apoptosis of lymphoid cells.

PubMed

Zhang, Lingzhi; Insel, Paul A

2004-05-14

The mechanisms by which cAMP mediates apoptosis are not well understood. In the current studies, we used wild-type (WT) S49 T-lymphoma cells and the kin(-) variant (which lacks protein kinase A (PKA)) to examine cAMP/PKA-mediated apoptosis. The cAMP analog, 8-CPT-cAMP, increased phosphorylation of the cAMP response element-binding protein (CREB), activated caspase-3, and induced apoptosis in WT but not in kin(-) S49 cells. Using an array of 96 apoptosis-related genes, we found that treatment of WT cells with 8-CPT-cAMP for 24 h induced expression of mRNA for the pro-apoptotic gene, Bim. Real-time PCR analysis indicated that 8-CPT-cAMP increased Bim RNA in WT cells in <2 h and maintained this increase for >24 h. Bim protein expression increased in WT but not kin(-) cells treated with 8-CPT-cAMP or with the beta-adrenergic receptor agonist isoproterenol. Both apoptosis and Bim expression were reversible with removal of 8-CPT-cAMP after <6 h. The glucocorticoid dexamethasone also promoted apoptosis and Bim expression in S49 cells. In contrast, both UV light and anti-mouse Fas monoclonal antibody promoted apoptosis in S49 cells but did not induce Bim expression. 8-CPT-cAMP also induced Bim expression and enhanced dexamethasone-promoted apoptosis in human T-cell leukemia CEM-C7-14 (glucocorticoid-sensitive) and CEM-C1-15 (glucocorticoid-resistant) cells; increased Bim expression in 8-CPT-cAMP-treated CEM-C1-15 cells correlated with conversion of the cells from resistance to sensitivity to glucocorticoid-promoted apoptosis. Induction of Bim appears to be a key event in cAMP-promoted apoptosis in both murine and human T-cell lymphoma and leukemia cells and thus appears to be a convergence point for the killing of such cells by glucocorticoids and agents that elevate cAMP.

Fibroblast growth factor rescues brain endothelial cells lacking presenilin 1 from apoptotic cell death following serum starvation.

PubMed

Gama Sosa, Miguel A; De Gasperi, Rita; Hof, Patrick R; Elder, Gregory A

2016-07-22

Presenilin 1 (Psen1) is important for vascular brain development and is known to influence cellular stress responses. To understand the role of Psen1 in endothelial stress responses, we investigated the effects of serum withdrawal on wild type (wt) and Psen1-/- embryonic brain endothelial cells. Serum starvation induced apoptosis in Psen1-/- cells but did not affect wt cells. PI3K/AKT signaling was reduced in serum-starved Psen1-/- cells, and this was associated with elevated levels of phospho-p38 consistent with decreased pro-survival AKT signaling in the absence of Psen1. Fibroblast growth factor (FGF1 and FGF2), but not vascular endothelial growth factor (VEGF) rescued Psen1-/- cells from serum starvation induced apoptosis. Inhibition of FGF signaling induced apoptosis in wt cells under serum withdrawal, while blocking γ-secretase activity had no effect. In the absence of serum, FGF2 immunoreactivity was distributed diffusely in cytoplasmic and nuclear vesicles of wt and Psen1-/- cells, as levels of FGF2 in nuclear and cytosolic fractions were not significantly different. Thus, sensitivity of Psen1-/- cells to serum starvation is not due to lack of FGF synthesis but likely to effects of Psen1 on FGF release onto the cell surface and impaired activation of the PI3K/AKT survival pathway.

FOXOs modulate proteasome activity in human-induced pluripotent stem cells of Huntington's disease and their derived neural cells.

PubMed

Liu, Yanying; Qiao, Fangfang; Leiferman, Patricia C; Ross, Alan; Schlenker, Evelyn H; Wang, Hongmin

2017-11-15

Although it has been speculated that proteasome dysfunction may contribute to the pathogenesis of Huntington's disease (HD), a devastating neurodegenerative disorder, how proteasome activity is regulated in HD affected stem cells and somatic cells remains largely unclear. To better understand the pathogenesis of HD, we analyzed proteasome activity and the expression of FOXO transcription factors in three wild-type (WT) and three HD induced-pluripotent stem cell (iPSC) lines. HD iPSCs exhibited elevated proteasome activity and higher levels of FOXO1 and FOXO4 proteins. Knockdown of FOXO4 but not FOXO1 expression decreased proteasome activity. Following neural differentiation, the HD-iPSC-derived neural progenitor cells (NPCs) demonstrated lower levels of proteasome activity and FOXO expressions than their WT counterparts. More importantly, overexpression of FOXO4 but not FOXO1 in HD NPCs dramatically enhanced proteasome activity. When HD NPCs were further differentiated into DARPP32-positive neurons, these HD neurons were more susceptible to death than WT neurons and formed Htt aggregates under the condition of oxidative stress. Similar to HD NPCs, HD-iPSC-derived neurons showed reduced proteasome activity and diminished FOXO4 expression compared to WT-iPSC-derived neurons. Furthermore, HD iPSCs had lower AKT activities than WT iPSCs, whereas the neurons derived from HD iPSC had higher AKT activities than their WT counterparts. Inhibiting AKT activity increased both FOXO4 level and proteasome activity, indicating a potential role of AKT in regulating FOXO levels. These data suggest that FOXOs modulate proteasome activity, and thus represents a potentially valuable therapeutic target for HD. © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Lack of effect of the alpha2C-adrenoceptor Del322-325 polymorphism on inhibition of cyclic AMP production in HEK293 cells.

PubMed

Montgomery, M D; Bylund, D B

2010-02-01

The alpha(2C)-adrenoceptor has multiple functions, including inhibiting release of noradrenaline from presynaptic nerve terminals. A human alpha(2C) polymorphism, Del322-325, a potential risk factor for heart failure, has been reported to exhibit reduced signalling in CHO cells. To further understand the role of the Del322-325 polymorphism on receptor signalling, we attempted to replicate and further study the reduced signalling in HEK293 cells. Human alpha(2C) wild-type (WT) and Del322-325 adrenoceptors were stably transfected into HEK293 cells. Radioligand binding was performed to determine affinities for both receptors. In intact cells, inhibition of forskolin-stimulated cyclic AMP production by WT and Del322-325 clones with a range of receptor densities (200-2320 fmol.mg(-1) protein) was measured following agonist treatment. Noradrenaline, brimonidine and clonidine exhibited similar binding affinities for WT and Del322-325. Brimonidine and clonidine also had similar efficacies and potencies for both receptors for the inhibition of cyclic AMP production at all receptor densities tested. A linear regression analysis comparing efficacy and potency with receptor expression levels showed no differences in slopes between WT and Del322-325. The alpha(2C) WT and Del322-325 adrenoceptors exhibited similar binding properties. Additionally, inhibition of cyclic AMP production by Del322-325 was similar to that of WT over a range of receptor densities. Therefore, in intact HEK293 cells, the alpha(2C)-Del322-325 polymorphism does not exhibit reduced signalling to adenylyl cyclase and may not represent a clinically important phenotype.

Bone marrow cell migration to the heart in a chimeric mouse model of acute chagasic disease

PubMed Central

Irion, Camila Iansen; Paredes, Bruno Diaz; Brasil, Guilherme Visconde; da Cunha, Sandro Torrentes; Paula, Luis Felipe; Carvalho, Alysson Roncally; de Carvalho, Antonio Carlos Campos; Carvalho, Adriana Bastos; Goldenberg, Regina Coeli dos Santos

2017-01-01

BACKGROUND Chagas disease is a public health problem caused by infection with the protozoan Trypanosoma cruzi. There is currently no effective therapy for Chagas disease. Although there is some evidence for the beneficial effect of bone marrow-derived cells in chagasic disease, the mechanisms underlying their effects in the heart are unknown. Reports have suggested that bone marrow cells are recruited to the chagasic heart; however, studies using chimeric mouse models of chagasic cardiomyopathy are rare. OBJECTIVES The aim of this study was to investigate the migration of bone marrow cells to the heart after T. cruzi infection in a model of chagasic disease in chimeric mice. METHODS To obtain chimerical mice, wild-type (WT) C57BL6 mice were exposed to full body irradiation (7 Gy), causing bone marrow ablation. Then, bone marrow cells from green fluorescent protein (GFP)-transgenic mice were infused into the mice. Graft effectiveness was confirmed by flow cytometry. Experimental mice were divided into four groups: (i) infected chimeric (iChim) mice; (ii) infected WT (iWT) mice, both of which received 3 × 104 trypomastigotes of the Brazil strain; (iii) non-infected chimeric (Chim) mice; and (iv) non-infected WT mice. FINDINGS At one-month post-infection, iChim and iWT mice showed first degree atrioventricular block with decreased heart rate and treadmill exercise parameters compared to those in the non-infected groups. MAIN CONCLUSIONS iChim mice showed an increase in parasitaemia, myocarditis, and the presence of amastigote nests in the heart tissue compared to iWT mice. Flow cytometry analysis did not detect haematopoietic progenitor cells in the hearts of infected mice. Furthermore, GFP+ cardiomyocytes were not detected in the tissues of chimeric mice. PMID:28767980

Bone marrow cell migration to the heart in a chimeric mouse model of acute chagasic disease.

PubMed

Irion, Camila Iansen; Paredes, Bruno Diaz; Brasil, Guilherme Visconde; Cunha, Sandro Torrentes da; Paula, Luis Felipe; Carvalho, Alysson Roncally; Carvalho, Antonio Carlos Campos de; Carvalho, Adriana Bastos; Goldenberg, Regina Coeli Dos Santos

2017-08-01

Chagas disease is a public health problem caused by infection with the protozoan Trypanosoma cruzi. There is currently no effective therapy for Chagas disease. Although there is some evidence for the beneficial effect of bone marrow-derived cells in chagasic disease, the mechanisms underlying their effects in the heart are unknown. Reports have suggested that bone marrow cells are recruited to the chagasic heart; however, studies using chimeric mouse models of chagasic cardiomyopathy are rare. The aim of this study was to investigate the migration of bone marrow cells to the heart after T. cruzi infection in a model of chagasic disease in chimeric mice. To obtain chimerical mice, wild-type (WT) C57BL6 mice were exposed to full body irradiation (7 Gy), causing bone marrow ablation. Then, bone marrow cells from green fluorescent protein (GFP)-transgenic mice were infused into the mice. Graft effectiveness was confirmed by flow cytometry. Experimental mice were divided into four groups: (i) infected chimeric (iChim) mice; (ii) infected WT (iWT) mice, both of which received 3 × 104 trypomastigotes of the Brazil strain; (iii) non-infected chimeric (Chim) mice; and (iv) non-infected WT mice. At one-month post-infection, iChim and iWT mice showed first degree atrioventricular block with decreased heart rate and treadmill exercise parameters compared to those in the non-infected groups. iChim mice showed an increase in parasitaemia, myocarditis, and the presence of amastigote nests in the heart tissue compared to iWT mice. Flow cytometry analysis did not detect haematopoietic progenitor cells in the hearts of infected mice. Furthermore, GFP+ cardiomyocytes were not detected in the tissues of chimeric mice.

Direct comparison of progenitor cells derived from adipose, muscle, and bone marrow from wild-type or craniosynostotic rabbits

PubMed Central

GM, Cooper; EL, Lensie; JJ, Cray; MR, Bykowski; GE, DeCesare; MA, Smalley; MP, Mooney; PG, Campbell; JE, Losee

2010-01-01

Background Reports have identified cells capable of osteogenic differentiation in bone marrow, muscle, and adipose tissues, but there are few direct comparisons of these different cell-types. Also, few have investigated the potential connection between a tissue-specific pathology and cells derived from seemingly unrelated tissues. Here, we compare cells isolated from wild-type rabbits or rabbits with nonsyndromic craniosynostosis, defined as the premature fusion of one or more of the cranial sutures. Methods Cells were derived from bone marrow, adipose, and muscle of 10 day-old wild-type rabbits (WT; n=17) or from age-matched rabbits with familial nonsyndromic craniosynostosis (CS; n=18). Cells were stimulated with bone morphogenetic protein 4 (BMP4) and alkaline phosphatase expression and cell proliferation were assessed. Results In WT rabbits, cells derived from muscle had more alkaline phosphatase activity than cells derived from either adipose or bone marrow. The cells derived from CS rabbit bone marrow and muscle were significantly more osteogenic than WT. Adipose-derived cells demonstrated no significant differences. While muscle-derived cells were most osteogenic in WT rabbits, bone marrow-derived cells were most osteogenic in CS rabbits. Conclusions Results suggest that cells from different tissues have different potentials for differentiation. Furthermore, cells derived from rabbits with craniosynostosis were different from wild-type derived cells. Interestingly, cells derived from the craniosynostotic rabbits were not uniformly more responsive compared with wild-type cells, suggesting that specific tissue-derived cells may react differently in individuals with craniosynostosis. PMID:20871482

Mesenchymal stem cells-derived MFG-E8 accelerates diabetic cutaneous wound healing.

PubMed

Uchiyama, Akihiko; Motegi, Sei-Ichiro; Sekiguchi, Akiko; Fujiwara, Chisako; Perera, Buddhini; Ogino, Sachiko; Yokoyama, Yoko; Ishikawa, Osamu

2017-06-01

Diabetic wounds are intractable due to complex factors, such as the inhibition of angiogenesis, dysfunction of phagocytosis by macrophages and abnormal inflammatory responses. It is recognized that mesenchymal stem cells (MSCs) promote wound healing in diabetic mice. We previously demonstrated that MSCs produce large amounts of MFG-E8. The objective was to ascertain the role of MSCs-derived MFG-E8 in murine diabetic wounds. MFG-E8 WT/KO MSCs or rMFG-E8 were subcutaneously injected around the wound in diabetic db/db mice, and wound areas were analyzed. Quantification of angiogenesis, infiltrating inflammatory cells, apoptotic cells at the wound area was performed by immunofluorescence staining and real-time PCR. Phagocytosis assay was performed using peritoneal macrophages from WT or db/db mice. MFG-E8 expression in granulation tissue in diabetic mice was significantly reduced compared with that in non-diabetic mice. We next examined the effect of subcutaneous injection of MFG-E8 WT/KO MSCs around the wound. Diabetic wound healing was significantly accelerated by the injection of MSCs. Diabetic wound healing in MFG-E8 KO MSCs-injected wounds was significantly delayed compared to that in WT MSCs-injected wounds. The numbers of CD31 + EC and NG2 + pericytes, as well as M2 macrophages in wounds in KO MSCs-injected mice were significantly decreased. MFG-E8 WT MSCs treatment suppressed the number of apoptotic cells and TNF-α + cells in wounds. In an in vitro assay, MFG-E8 WT MSCs-conditioned medium enhanced phagocytosis of apoptotic cells by peritoneal macrophages from diabetic mice. MSCs-derived MFG-E8 might accelerate diabetic wound healing by promoting angiogenesis, the clearance of apoptotic cells, and the infiltration of M2 macrophages, and by suppressing inflammatory cytokines in wound area. Copyright © 2017 Japanese Society for Investigative Dermatology. Published by Elsevier B.V. All rights reserved.

Characterization of the molecular distribution of drugs in glassy solid dispersions at the nano-meter scale, using differential scanning calorimetry and gravimetric water vapour sorption techniques.

PubMed

van Drooge, D J; Hinrichs, W L J; Visser, M R; Frijlink, H W

2006-03-09

The molecular distribution in fully amorphous solid dispersions consisting of poly(vinylpyrrolidone) (PVP)-diazepam and inulin-diazepam was studied. One glass transition temperature (T(g)), as determined by temperature modulated differential scanning calorimetry (TMDSC), was observed in PVP-diazepam solid dispersions prepared by fusion for all drug loads tested (10-80 wt.%). The T(g) of these solid dispersions gradually changed with composition and decreased from 177 degrees C for pure PVP to 46 degrees C for diazepam. These observations indicate that diazepam was dispersed in PVP on a molecular level. However, in PVP-diazepam solid dispersions prepared by freeze drying, two T(g)'s were observed for drug loads above 35 wt.% indicating phase separation. One T(g) indicated the presence of amorphous diazepam clusters, the other T(g) was attributed to a PVP-rich phase in which diazepam was dispersed on a molecular level. With both the value of the latter T(g) and the DeltaC(p) of the diazepam glass transition the concentrations of molecular dispersed diazepam could be calculated (27-35 wt.%). Both methods gave similar results. Water vapour sorption (DVS) experiments revealed that the PVP-matrix was hydrophobised by the incorporated diazepam. TMDSC and DVS results were used to estimate the size of diazepam clusters in freeze dried PVP-diazepam solid dispersions, which appeared to be in the nano-meter range. The inulin-diazepam solid dispersions prepared by spray freeze drying showed one T(g) for drug loads up to 35 wt.% indicating homogeneous distribution on a molecular level. However, this T(g) was independent of the drug load, which is unexpected because diazepam has a lower T(g) than inulin (46 and 155 degrees C, respectively). For higher drug loads, a T(g) of diazepam as well as a T(g) of the inulin-rich phase was observed, indicating the formation of amorphous diazepam clusters. From the DeltaC(p) of the diazepam glass transition the amount of molecularly dispersed diazepam was calculated (12-27 wt.%). In contrast to the PVP-diazepam solid dispersions, DVS-experiments revealed that inulin was not hydrophobised by diazepam. Consequently, the size of diazepam clusters could not be estimated. It was concluded that TMDSC enables characterization and quantification of the molecular distribution in amorphous solid dispersions. When the hygroscopicity of the carrier is reduced by the drug, DVS in combination with TMDSC can be used to estimate the size of amorphous drug clusters.

Unraveling the catalyzing behaviors of different iron species (Fe2+ vs. Fe0) in activating persulfate-based oxidation process with implications to waste activated sludge dewaterability.

PubMed

Zhen, Guangyin; Lu, Xueqin; Su, Lianghu; Kobayashi, Takuro; Kumar, Gopalakrishnan; Zhou, Tao; Xu, Kaiqin; Li, Yu-You; Zhu, Xuefeng; Zhao, Youcai

2018-05-01

Dewatering of waste activated sludge (WAS) is of major interest in its volume reduction, transportation and ultimate disposal. Persulfate-based oxidation process is a newly developed option for enhancing WAS dewaterability through the generation of powerful sulfate radicals (SO 4 - ·). However, the enhancement in WAS dewaterability by persulfate differs with the species of iron catalysts used. In this study, two types of iron catalysts (i.e. Fe 2+ vs. Fe 0 ) were employed to initiate the persulfate (S 2 O 8 2- ), and the catalyzing behaviors and the underlying principles in enhancing WAS dewaterability were investigated and compared. The Fe 2+ exhibited the high effectiveness in catalyzing the decomposition of persulfate to sulfate radicals (SO 4 - ·), inducing the greater improvement in WAS dewatering. The WAS dewaterability (indicated by dry solids content after filtration) increased with the added S 2 O 8 2- /Fe 2+ dosages, with the dry solids content reaching up to 5.1 ± 0.8 wt% at S 2 O 8 2- /Fe 2+ dosages of 1.2/1.5 mmol/g-VS after only 30 s' filtration, roughly 1.8-fold increase than raw WAS (1.8 ± 0.1 wt%). In contrast, the influence of the persulfate oxidation when activated with Fe 0 on WAS dewaterability was statistically insignificant. The WAS dewaterability remained nearly unchanged (i.e. dry solids content of 2.0 ± 0.0 wt%), irrespective of the employed S 2 O 8 2- /Fe 0 dosages. Further analysis demonstrated that the WAS dewaterability negatively corresponded to loosely bound extracellular polymeric substances (LB-EPS) and tightly bound EPS (TB-EPS). The abundant SO 4 - · from S 2 O 8 2- /Fe 2+ system could effectively disrupt the gel-like EPS matrix, break apart the cells and subsequently arouse the release of the water inside EPS and cells, facilitating water-solid separation. In the case of S 2 O 8 2- /Fe 0 , the dissolution of Fe 0 particles was the rate-limiting step, due to the formation of oxide iron layer near Fe 0 metallic surface, which resulted in the slow SO 4 - · production and thus hardly promoted WAS dewaterability. The pH adjustment could accelerate Fe 0 dissolution and enhance the dewatering performance of S 2 O 8 2- /Fe 0 process to a certain degree, but the effect was unsatisfactory. Additionally, the observations regarding the dissolved organic matters and ammonium collectively revealed that except for enhancing WAS dewatering, S 2 O 8 2- /Fe 2+ oxidation could concurrently degrade COD and ammonia from WAS filtrate, lighten the burden of the subsequent sewage treatment facilities and reduce operational expense. Hence, from an environmental and economic perspective, the S 2 O 8 2- /Fe 2+ system possesses much greater promise for WAS dewatering. Copyright © 2018 Elsevier Ltd. All rights reserved.

CAR-Engineered NK Cells Targeting Wild-Type EGFR and EGFRvIII Enhance Killing of Glioblastoma and Patient-Derived Glioblastoma Stem Cells.

PubMed

Han, Jianfeng; Chu, Jianhong; Keung Chan, Wing; Zhang, Jianying; Wang, Youwei; Cohen, Justus B; Victor, Aaron; Meisen, Walter H; Kim, Sung-hak; Grandi, Paola; Wang, Qi-En; He, Xiaoming; Nakano, Ichiro; Chiocca, E Antonio; Glorioso, Joseph C; Kaur, Balveen; Caligiuri, Michael A; Yu, Jianhua

2015-07-09

Glioblastoma (GB) remains the most aggressive primary brain malignancy. Adoptive transfer of chimeric antigen receptor (CAR)-modified immune cells has emerged as a promising anti-cancer approach, yet the potential utility of CAR-engineered natural killer (NK) cells to treat GB has not been explored. Tumors from approximately 50% of GB patients express wild-type EGFR (wtEGFR) and in fewer cases express both wtEGFR and the mutant form EGFRvIII; however, previously reported CAR T cell studies only focus on targeting EGFRvIII. Here we explore whether both wtEGFR and EGFRvIII can be effectively targeted by CAR-redirected NK cells to treat GB. We transduced human NK cell lines NK-92 and NKL, and primary NK cells with a lentiviral construct harboring a second generation CAR targeting both wtEGFR and EGFRvIII and evaluated the anti-GB efficacy of EGFR-CAR-modified NK cells. EGFR-CAR-engineered NK cells displayed enhanced cytolytic capability and IFN-γ production when co-cultured with GB cells or patient-derived GB stem cells in an EGFR-dependent manner. In two orthotopic GB xenograft mouse models, intracranial administration of NK-92-EGFR-CAR cells resulted in efficient suppression of tumor growth and significantly prolonged the tumor-bearing mice survival. These findings support intracranial administration of NK-92-EGFR-CAR cells represents a promising clinical strategy to treat GB.

Effect of Few-Layered Graphene-Based CdO Nanocomposite-Enhanced Power Conversion Efficiency of Dye-Sensitized Solar Cell

NASA Astrophysics Data System (ADS)

Bykkam, Satish; Kalagadda, Bikshalu; Kalagadda, Venkateswara Rao; Ahmadipour, Mohsen; Chakra, Ch. Shilpa; Rajendar, V.

2018-01-01

A few-layered graphene (FLG)/cadmium oxide (CdO) nanocomposite was sucessfully prepared through ultrasonic-assisted synthesis. The morphology of FLG (1.0 wt.%, 2.0 wt.%, and 3.0 wt.%)/CdO nanocomposites were characterized using high-resolution transmission electron microscopy and field emission scanning electron microscopy techniques. The optical properties were studied with the help of UV-Vis diffuse reflectance spectroscopy and Raman spectroscopy, while the crystalline phases were analyzed using x-ray diffraction. The doctor blade method was used to deposit FLG/CdO nanocomposites on fluorine-doped tin oxide conductive glass substrates. The effect of FLG weight percentage (1.0 wt.%, 2.0 wt.%, and 3.0 wt.%) was studied on the power conversion efficiency of dye-sensitized solar cell applications. The photovoltaic characteristics, current density-voltage curves were measured with ruthenium (II)-based dye under air mass condition 1.5G, 100 m W m-2 of a solar simulator. The results showed that higher power conversion efficiency of 3.54% was achieved at the appropriate weight percentage of FLG (1.0 wt.%)/CdO nanocomposite, compared to the CdO and other nanocomposite working electrodes FLG (2.0 wt.%, and 3.0 wt.%)/CdO.

Somato-synaptic variation of GABA(A) receptors in cultured murine cerebellar granule cells: investigation of the role of the alpha6 subunit.

PubMed

Mellor, J R; Wisden, W; Randall, A D

2000-07-10

Electrophysiological investigation of cultured cerebellar murine granule cells revealed differences between the GABA(A) receptors at inhibitory synapses and those on the cell body. Specifically, mIPSCs decayed more rapidly than cell body receptors deactivated, the mean single channel conductance at the synapse (32 pS) was greater than that at cell body (21 pS) and only cell body receptors were sensitive to Zn(2+) (150 microM), which depressed response amplitude by 82+/-5% and almost doubled the rate of channel deactivation. The GABA(A) receptor alpha6 subunit is selectively expressed in cerebellar granule cells. Although concentrated at synapses, it is also found on extrasynaptic membranes. Using a mouse line (Deltaalpha6lacZ) lacking this subunit, we investigated its role in the somato-synaptic differences in GABA(A) receptor function. All differences between cell body and synaptic GABA(A) receptors observed in wild-type (WT) granule cells persisted in Deltaalpha6lacZ cells, thus demonstrating that they are not specifically due to the cellular distribution of the alpha6 subunit. However, mIPSCs from WT and Deltaalpha6lacZ cells differed in both their kinetics (faster decay in WT cells) and underlying single channel conductance (32 pS WT, 25 pS Deltaalpha6lacZ). This provides good evidence for a functional contribution of the alpha6 subunit to postsynaptic GABA(A) receptors in these cells. Despite this, deactivation kinetics of mIPSCs in WT and Deltaalpha6lacZ granule cells exhibited similar benzodiazepene (BDZ) sensitivity. This suggests that the enhanced BDZ-induced ataxia seen in Deltaalpha6lacZ mice may reflect physiological activity at extrasynaptic receptors which, unlike those at synapses, display differential BDZ-sensitivity in WT and Deltaalpha6lacZ granule cells (Jones, A.M., Korpi, E.R., McKernan, R.M., Nusser, Z., Pelz, R., Makela, R., Mellor, J.R., Pollard, S., Bahn, S., Stephenson, F.A., Randall, A.D., Sieghart, W., Somogyi, P., Smith, A.J.H., Wisden, W., 1997. Ligand-gated ion channel partnerships: GABA(A) receptor alpha(6) subunit inactivation inhibits delta subunit expression. Journal of Neuroscience 17, 1350-1362).

In Vivo Assays for Assessing the Role of the Wilms' Tumor Suppressor 1 (Wt1) in Angiogenesis.

PubMed

McGregor, Richard J; Ogley, R; Hadoke, Pwf; Hastie, Nicholas

2016-01-01

The Wilms' tumor suppressor gene (WT1) is widely expressed during neovascularization, but it is almost entirely absent in quiescent adult vasculature. However, in vessels undergoing angiogenesis, WT1 is dramatically upregulated. Studies have shown Wt1 has a role in both tumor and ischemic angiogenesis, but the mechanism of Wt1 action in angiogenic tissue remains to be elucidated. Here, we describe two methods for induction of in vivo angiogenesis (subcutaneous sponge implantation, femoral artery ligation) that can be used to assess the influence of Wt1 on new blood vessel formation. Subcutaneously implanted sponges stimulate an inflammatory and fibrotic response including cell infiltration and angiogenesis. Femoral artery ligation creates ischemia in the distal hindlimb and produces an angiogenic response to reperfuse the limb which can be quantified in vivo by laser Doppler flowmetry. In both of these models, the role of Wt1 in the angiogenic process can be assessed using histological/immunohistochemical staining, molecular analysis (qPCR) and flow cytometry. Furthermore, combined with suitable genetic modifications, these models can be used to explore the causal relationship between Wt1 expression and angiogenesis and to trace the lineage of cells expressing Wt1. This approach will help to clarify the importance of Wt1 in regulating neovascularization in the adult, and its potential as a therapeutic target.

Reactivation of wild-type and mutant p53 by tryptophanolderived oxazoloisoindolinone SLMP53-1, a novel anticancer small-molecule

PubMed Central

Soares, Joana; Raimundo, Liliana; Pereira, Nuno A.L.; Monteiro, Ângelo; Gomes, Sara; Bessa, Cláudia; Pereira, Clara; Queiroz, Glória; Bisio, Alessandra; Fernandes, João; Gomes, Célia; Reis, Flávio; Gonçalves, Jorge; Inga, Alberto; Santos, Maria M.M.; Saraiva, Lucília

2016-01-01

Restoration of the p53 pathway, namely by reactivation of mutant (mut) p53, represents a valuable anticancer strategy. Herein, we report the identification of the enantiopure tryptophanol-derived oxazoloisoindolinone SLMP53-1 as a novel reactivator of wild-type (wt) and mut p53, using a yeast-based screening strategy. SLMP53-1 has a p53-dependent anti-proliferative activity in human wt and mut p53R280K-expressing tumor cells. Additionally, SLMP53-1 enhances p53 transcriptional activity and restores wt-like DNA binding ability to mut p53R280K. In wt/mut p53-expressing tumor cells, SLMP53-1 triggers p53 transcription-dependent and mitochondrial apoptotic pathways involving BAX, and wt/mut p53 mitochondrial translocation. SLMP53-1 inhibits the migration of wt/mut p53-expressing tumor cells, and it shows promising p53-dependent synergistic effects with conventional chemotherapeutics. In xenograft mice models, SLMP53-1 inhibits the growth of wt/mut p53-expressing tumors, but not of p53-null tumors, without apparent toxicity. Collectively, besides the potential use of SLMP53-1 as anticancer drug, the tryptophanol-derived oxazoloisoindolinone scaffold represents a promissing starting point for the development of effective p53-reactivating drugs. PMID:26735173

Influences of CdSe NCs on the photovoltaic parameters of BHJ organic solar cells

NASA Astrophysics Data System (ADS)

Ongul, Fatih; Yuksel, Sureyya Aydin; Allahverdi, Cagdas; Bozar, Sinem; Kazici, Mehmet; Gunes, Serap

2018-04-01

In this study, the high quality CdSe nanocrystals (NCs) capped with stearic acid were synthesized in a solvent and then purified four times by using the precipitation and redissolution process. The average size of the synthesized CdSe NCs was determined 3.0 nm via transmission electron microscopy (TEM) measurement and their corresponding optical band edge energy was also calculated as 2.1 eV using ultraviolet-visible (UV-Vis) absorption spectroscopy. The bulk heterojunction (BHJ) hybrid solar cells based on a ternary system including P3HT, PCBM and CdSe NCs at different weight concentrations (0 wt%, 0.1 wt%, 0.5 wt%, 1 wt% and 2 wt%) were fabricated by spin-casting process. The effect of the concentration of CdSe NCs on the photovoltaic parameters of these BHJ organic solar cells was investigated. The surface morphology of the photoactive layer modified by the incorporation of CdSe NCs into P3HT:PCBM matrix was observed with scanning electron microscopy (SEM). It was shown that when the concentration of CdSe NCs increases above 0.1 wt% in this ternary system, the photovoltaic performance of the devices significantly decreases. The power conversion efficiency of the organic photovoltaic (OPV) device was enhanced 20% by incorporating CdSe NCs with 0.1 wt% with respect to those without CdSe NCs.

Influences of CdSe NCs on the photovoltaic parameters of BHJ organic solar cells.

PubMed

Ongul, Fatih; Yuksel, Sureyya Aydin; Allahverdi, Cagdas; Bozar, Sinem; Kazici, Mehmet; Gunes, Serap

2018-04-05

In this study, the high quality CdSe nanocrystals (NCs) capped with stearic acid were synthesized in a solvent and then purified four times by using the precipitation and redissolution process. The average size of the synthesized CdSe NCs was determined ~3.0nm via transmission electron microscopy (TEM) measurement and their corresponding optical band edge energy was also calculated as ~2.1eV using ultraviolet-visible (UV-Vis) absorption spectroscopy. The bulk heterojunction (BHJ) hybrid solar cells based on a ternary system including P3HT, PCBM and CdSe NCs at different weight concentrations (0wt%, 0.1wt%, 0.5wt%, 1wt% and 2wt%) were fabricated by spin-casting process. The effect of the concentration of CdSe NCs on the photovoltaic parameters of these BHJ organic solar cells was investigated. The surface morphology of the photoactive layer modified by the incorporation of CdSe NCs into P3HT:PCBM matrix was observed with scanning electron microscopy (SEM). It was shown that when the concentration of CdSe NCs increases above 0.1wt% in this ternary system, the photovoltaic performance of the devices significantly decreases. The power conversion efficiency of the organic photovoltaic (OPV) device was enhanced ~20% by incorporating CdSe NCs with 0.1wt% with respect to those without CdSe NCs. Copyright © 2017 Elsevier B.V. All rights reserved.

Supercritical water pyrolysis of sewage sludge.

PubMed

Ma, Wenchao; Du, Guiyue; Li, Jian; Fang, Yuanhao; Hou, Li'an; Chen, Guanyi; Ma, Degang

2017-01-01

Municipal sewage sludge (SS) from wastewater treatment plant containing high water content (>85wt.%), lead to the difficulty of co-combustion with MSW or coal due to the high cost of drying. This study explores an alternative method by supercritical water (SCW) pyrolysis of sewage sludge (SS) in a high pressure reaction vessel. The effects of temperature and moisture content of SS on yield and composition of the products (bio-oil, bio char and non-condensable gas) were studied. A temperature of 385°C and moisture content of 85wt.% were found to be the optimum conditions for the maximum bio-oil production of 37.23wt.%, with a higher heating value of 31.08MJ/kg. In the optimum condition, the yields of aliphatic hydrocarbon and phenols were about 29.23wt.% and 12.51wt.%, respectively. The physical and chemical properties of bio-char were analyzed by using XRF and BET. Results of GC analyses of NCG showed that it has the maximum HHV of 13.39MJ/m 3 at 445°C and moisture content of 85wt.%. The reaction path from SS to bio-oil through SCW pyrolysis was given. Moreover, carbon balance was calculated for the optimal condition, and finding out that 64.27wt.% of the carbon content was transferred from SS to bio-oil. Finally, this work demonstrates that the SCW pyrolysis is a promising disposal method for SS. Copyright © 2016. Published by Elsevier Ltd.

IDH1 R132H mutation regulates glioma chemosensitivity through Nrf2 pathway.

PubMed

Li, Kaishu; Ouyang, Leping; He, Mingliang; Luo, Ming; Cai, Wangqing; Tu, Yalin; Pi, Rongbiao; Liu, Anmin

2017-04-25

Numerous studies have reported that glioma patients with isocitrate dehydrogenase 1(IDH1) R132H mutation are sensitive to temozolomide treatment. However, the mechanism of IDH1 mutations on the chemosensitivity of glioma remains unclear. In this study, we investigated the role and the potential mechanism of Nrf2 in IDH1 R132H-mediated drug resistance. Wild type IDH1 (R132H-WT) and mutant IDH1 (R132H) plasmids were constructed. Stable U87 cells and U251 cells overexpressing IDH1 were generated. Phenotypic differences between IDH1-WT and IDH1 R132H overexpressing cells were evaluated using MTT, cell colony formation assay, scratch test assay and flow cytometry. Expression of IDH1 and its associated targets, nuclear factor-erythroid 2-related factor 2 (Nrf2), NAD(P)H quinine oxidoreductase 1 (NQO1), multidrug resistant protein 1 (MRP1) and p53 were analyzed. The IDH1 R132H overexpressing cells were more sensitive to temozolomide than WT and the control, and Nrf2 was significantly decreased in IDH1 R132H overexpressing cells. We found that knocking down Nrf2 could decrease resistance to temozolomide. The nuclear translocation of Nrf2 in IDH1 R132H overexpressing cells was lower than the WT and the control groups after temozolomide treatment. When compared with WT cells, NQO1 expression was reduced in IDH1 R132H cells, especially after temozolomide treatment. P53 was involved in the resistance mechanism of temozolomide mediated by Nrf2 and NQO1. Nrf2 played an important role in IDH1 R132H-mediated drug resistance. The present study provides new insight for glioma chemotherapy with temozolomide.

IDH1 R132H mutation regulates glioma chemosensitivity through Nrf2 pathway

PubMed Central

Luo, Ming; Cai, Wangqing; Tu, Yalin; Pi, Rongbiao; Liu, Anmin

2017-01-01

Purpose Numerous studies have reported that glioma patients with isocitrate dehydrogenase 1(IDH1) R132H mutation are sensitive to temozolomide treatment. However, the mechanism of IDH1 mutations on the chemosensitivity of glioma remains unclear. In this study, we investigated the role and the potential mechanism of Nrf2 in IDH1 R132H-mediated drug resistance. Methods Wild type IDH1 (R132H-WT) and mutant IDH1 (R132H) plasmids were constructed. Stable U87 cells and U251 cells overexpressing IDH1 were generated. Phenotypic differences between IDH1-WT and IDH1 R132H overexpressing cells were evaluated using MTT, cell colony formation assay, scratch test assay and flow cytometry. Expression of IDH1 and its associated targets, nuclear factor-erythroid 2-related factor 2 (Nrf2), NAD(P)H quinine oxidoreductase 1 (NQO1), multidrug resistant protein 1 (MRP1) and p53 were analyzed. Results The IDH1 R132H overexpressing cells were more sensitive to temozolomide than WT and the control, and Nrf2 was significantly decreased in IDH1 R132H overexpressing cells. We found that knocking down Nrf2 could decrease resistance to temozolomide. The nuclear translocation of Nrf2 in IDH1 R132H overexpressing cells was lower than the WT and the control groups after temozolomide treatment. When compared with WT cells, NQO1 expression was reduced in IDH1 R132H cells, especially after temozolomide treatment. P53 was involved in the resistance mechanism of temozolomide mediated by Nrf2 and NQO1. Conclusions Nrf2 played an important role in IDH1 R132H-mediated drug resistance. The present study provides new insight for glioma chemotherapy with temozolomide. PMID:28427200

Long-term stability of dental adhesive incorporated by boron nitride nanotubes.

PubMed

Degrazia, Felipe Weidenbach; Leitune, Vicente Castelo Branco; Visioli, Fernanda; Samuel, Susana Maria Werner; Collares, Fabrício Mezzomo

2018-03-01

The aim of this study was to evaluate physicochemical properties, long-term microtensile bond strength and cytotoxicity of methacrylate-based adhesive containing boron nitride nanotubes (BNNTs) as fillers. A dental adhesive was formulated using BisGMA/HEMA, 66/33wt% (control). Inorganic BNNT fillers were incorporated into the adhesive at different concentrations (0.05, 0.075, 0.1 and 0.15wt%). Analyses of degree of conversion (DC), polymerization rate [Rp.(s -1 )], contact angle (CA) on dentin, after 24h and 6 months microtensile bond strength (μTBS-24h and 6 months) were assessed. Cytotoxicity was performed through viability of fibroblast cells (%) by sulforhodamine B (SRB) colorimetry. DC and max. polymerization rate increased (p<0.05) after incorporating 0.075 and 0.1wt% BNNT. The contact angle on dentin increased (p<0.05) after incorporating 0.15wt% BNNT. The μTBS-24h showed no changes (p>0.05) after incorporating up to 0.15wt% BNNT comparing to control. After 6 months, μTBS decreased (p<0.05) for control and 0.15wt% BNNT and BNNT groups up to 0.15wt% showed higher μTBS than control (p<0.05). No difference of fibroblast growth was found among adhesives (p>0.05) and up to 19% of cell viability was found comparing 0.05wt% BNNT to positive control group (100%). Incorporating boron nitride nanotubes up to 0.1wt% into dental adhesive increased the long-term stability to dentin without decreasing viability of fibroblast cell growth. Thus, the use of BNNTs as filler may decrease failure rate of current dentinal adhesives. Copyright © 2017 The Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

Roles of Alum and Monophosphoryl Lipid A Adjuvants in Overcoming CD4+ T Cell Deficiency to Induce Isotype-Switched IgG Antibody Responses and Protection by T-Dependent Influenza Vaccine

PubMed Central

Ko, Eun-Ju; Lee, Young-Tae; Kim, Ki-Hye; Lee, Youri; Jung, Yu-Jin; Kim, Min-Chul; Lee, Yu-Na; Kang, Taeuk; Kang, Sang-Moo

2016-01-01

Vaccine adjuvant effects in CD4 deficient condition largely remain unknown. We investigated the roles of combined monophosphoryl lipid A (MPL) and Alum adjuvant (MPL+Alum) in inducing immunity after immunization of CD4-knockout (CD4KO) and wild-type (WT) mice with T-dependent influenza vaccine. MPL+Alum adjuvant mediated IgG isotype-switched antibodies, IgG secreting cell responses, and protection in CD4KO mice, which were comparable to those in WT mice. In contrast, Alum adjuvant effects were dependent on CD4+ T cells. MPL+Alum adjuvant was effective in recruiting monocytes and neutrophils as well as in protecting macrophages from alum-mediated cell loss at the injection site in CD4KO mice. MPL+Alum appeared to attenuate MPL-induced inflammatory responses in WT mice, likely improving the safety. Additional studies in CD4-depleted WT mice and MHCII KO mice suggest that MHCII positive antigen presenting cells contribute to providing alternative B cell help in CD4 deficient condition in the context of MPL+Alum adjuvanted vaccination. PMID:27881702

High strength porous support tubes for high temperature solid electrolyte electrochemical cells

DOEpatents

Rossing, Barry R.; Zymboly, Gregory E.

1986-01-01

A high temperature, solid electrolyte electrochemical cell is made, having an electrode and a solid electrolyte disposed on a porous, sintered support material containing thermally stabilized zirconia powder particles and from about 3 wt. % to about 45 wt. % of thermally stable oxide fibers.

PRDM1 expression on the epithelial component but not on ectopic lymphoid tissues of Warthin tumour.

PubMed

Wang, Y; Zhou, J; Zhang, Y; Wang, L; Liu, Y; Fan, L; Zhu, J; Xu, X; Huang, G; Li, X; Xun, W

2015-05-01

To determine the role of PRDM1, a key molecule for modulating the immune cells, in Warthin tumour (WT) pathogenesis. Forty paraffin-embedded parotid tissues of patients (mean age: 62.08 ± 11.90) with WT were retrieved from the pathology archives of Qindu Hospital from January 2012 to December 2012. The PRDM1 expression was investigated in a cohort of WT by immunohistochemistry. PRDM1 was expressed only on the epithelial component but not on ectopic lymphoid tissue of the tumour. Statistically, PRDM1 expression rates between WT glandular epithelial cells (40/40 cases) and the tumour-adjacent tissues (0/9 cases), and WT germinal centres (0/34 cases) and tonsil tissues (10/10 cases) were significantly different (P < 0.001), respectively. The PRDM1 expression appeared to play an essential role in WT pathogenesis. A better understanding of it might give options for revealing possible novel management strategies. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Respiratory syncytial virus infection disrupts monolayer integrity and function in cystic fibrosis airway cells.

PubMed

Kong, Michele; Maeng, Patrick; Hong, Jeong; Szczesniak, Rhonda; Sorscher, Eric; Sullender, Wayne; Clancy, John Paul

2013-09-19

Respiratory Syncytial Virus (RSV) infection is a common contributor to pulmonary symptoms in children with cystic fibrosis (CF). Here we examined RSV infection in immortalized bronchial epithelial cells (CFBE41o-) expressing wild-type (wt) or F508del cystic fibrosis transmembrane conductance regulator (CFTR), for monolayer integrity and RSV replication. CFBE41o- monolayers expressing wt or F508del CFTR were grown on permeable supports and inoculated with RSV A2 strain. Control experiments utilized UV-inactivated RSV and heat-killed RSV. Monolayer resistance and RSV production was monitored for up to six days post-infection. Within 24 h, a progressive decrease in monolayer resistance was observed in RSV infected F508del CFBE41o- cells, while the monolayer integrity of RSV infected wt CFTR CFBE41o- cells remained stable. RSV replication was necessary to disrupt F508del CFBE41o- monolayers as UV-irradiated and heat killed RSV had no effect on monolayer integrity, with an earlier and much more pronounced peak in RSV titer noted in F508del relative to wt CFTR-expressing cells. RSV infection of wt CFBE41o- monolayers also resulted in blunting of CFTR response. These findings identify an enhanced sensitivity of CFBE41o- cells expressing F508del CFTR to RSV infection, replication and monolayer disruption independent of the cellular immune response, and provide a novel mechanism by which cystic fibrosis airway epithelia are susceptible to RSV-dependent injury.

Production, Survival, and Evaluation of Solid-Substrate Inocula of Penicillium oxalicum, a Biocontrol Agent Against Fusarium Wilt of Tomato.

PubMed

Larena, I; Melgarejo, P; De Cal, A

2002-08-01

ABSTRACT Production of conidia of Penicillium oxalicum (ATCC number pending), a biocontrol agent of Fusarium oxysporum f. sp. lycopersici, was tested in liquid and solid fermentation. P. oxalicum produced 250-fold more conidia in solid than in liquid fermentation at 30 days after inoculation of substrate. Solid fermentation was carried out in plastic bags (600 cm(3)) especially designed for solid fermentation (VALMIC) containing 50 g of peat/vermiculite (PV) (1:1, wt/wt) with 40% moisture, sealed, sterilized, and then inoculated with 1 ml of a conidial suspension of P. oxalicum (10(5) conidia g(-1) dry substrate), sealed again, and incubated in darkness at 20 to 25 degrees C for 30 days. Addition of amendments to PV in a proportion of 0.5 (wt/wt) significantly increased conidial production of P. oxalicum. The best production was obtained on PV plus meal of cereal grains (barley) or leguminous seeds (lentil) (100-fold higher). Conidial production obtained after 5 days of inoculation was similar to that obtained at 30 days. However, viability of conidia produced in PV plus lentil meal was 35% higher than that of conidia produced in PV plus barley meal. Changes in proportions (1:1:0.5, wt/wt/wt; 1:1:1, wt/wt/wt; 1:0.5:0.5, wt/wt/wt; 1:1:0.5, vol/vol/vol) of components of the substrate (peat/vermiculite/lentil meal) did not enhance production or viability of conidia. Optimal initial moisture in the substrate was 30 to 40%. At lower moistures, significant reductions of production of conidia were observed, particularly at 10%. There was a general decline in the number of conidia in bags with time of storage at -80, -20, 4, and 25 degrees C, or at room temperature (range from 30 to 15 degrees C), with the highest decline occurring from 60 to 180 days. Conidial viability also was reduced with time, except for conidia stored at -20 degrees C. Fresh conidia produced in solid fermentation system or those conidia stored at -20 degrees C for 180 days reduced Fusarium wilt of tomato by 49 and 61%, respectively.

Serotonin transporter protects the placental cells against apoptosis in caspase 3-independent pathway.

PubMed

Hadden, Coedy; Fahmi, Tariq; Cooper, Anthonya; Savenka, Alena V; Lupashin, Vladimir V; Roberts, Drucilla J; Maroteaux, Luc; Hauguel-de Mouzon, Sylvie; Kilic, Fusun

2017-12-01

Serotonin (5-HT) and its specific transporter, SERT play important roles in pregnancy. Using placentas dissected from 18d gestational SERT-knock out (KO), peripheral 5-HT (TPH1)-KO, and wild-type (WT) mice, we explored the role of 5-HT and SERT in placental functions in detail. An abnormal thick band of fibrosis and necrosis under the giant cell layer in SERT-KO placentas appeared only moderately in TPH1-KO and minimally present in WT placentas. The majority of the changes were located at the junctional zone of the placentas in SERT. The etiology of these findings was tested with TUNEL assays. The placentas from SERT-KO and TPH1-KO showed 49- and 8-fold increase in TUNEL-positive cells without a concurrent change in the DNA repair or cell proliferation compared to WT placentas. While the proliferation rate in the embryos of TPH1-KO mice was 16-fold lower than the rate in gestational age matched embryos of WT or SERT-KO mice. These findings highlight an important role of continuous 5-HT signaling on trophoblast cell viability. SERT may contribute to protecting trophoblast cells against cell death via terminating the 5-HT signaling which changes cell death ratio in trophoblast as well as proliferation rate in embryos. However, the cell death in SERT-KO placentas is in caspase 3-independent pathway. © 2017 Wiley Periodicals, Inc.

Creep of experimental short fiber-reinforced composite resin.

PubMed

Garoushi, Sufyan; Kaleem, Muhammad; Shinya, Akikazu; Vallittu, Pekka K; Satterthwaite, Julian D; Watts, David C; Lassila, Lippo V J

2012-01-01

The purpose of this study was to investigate the reinforcing effect of short E-glass fiber fillers oriented in different directions on composite resin under static and dynamic loading. Experimental short fiber-reinforced composite resin (FC) was prepared by mixing 22.5 wt% of short E-glass fibers, 22.5 wt% of resin, and 55 wt% of silane-treated silica fillers. Three groups of specimens (n=5) were tested: FC with isotropic fiber orientation, FC with anisotropic fiber orientation, and particulate-filled composite resin (PFC) as a control. Time-dependent creep and recovery were recorded. ANOVA revealed that after secondary curing in a vacuum oven and after storage in dry condition for 30 days, FC with isotropic fiber orientation (1.73%) exhibited significantly lower static creep value (p<0.05) than PFC (2.54%). For the different curing methods and storage conditions evaluated in this study, FC achieved acceptable static and dynamic creep values when compared to PFC.

CD4+ Foxp3+ T-cells contribute to myocardial ischemia-reperfusion injury.

PubMed

Mathes, Denise; Weirather, Johannes; Nordbeck, Peter; Arias-Loza, Anahi-Paula; Burkard, Matthias; Pachel, Christina; Kerkau, Thomas; Beyersdorf, Niklas; Frantz, Stefan; Hofmann, Ulrich

2016-12-01

The present study analyzed the effect of CD4 + Forkhead box protein 3 negative (Foxp3 - ) T-cells and Foxp3 + CD4 + T-cells on infarct size in a mouse myocardial ischemia-reperfusion model. We examined the infarct size as a fraction of the area-at-risk as primary study endpoint in mice after 30minutes of coronary ligation followed by 24hours of reperfusion. CD4 + T-cell deficient MHC-II KO mice showed smaller histologically determined infarct size (34.5±4.7% in MHCII KO versus 59.4±4.9% in wildtype (WT)) and better preserved ejection fraction determined by magnetic resonance tomography (56.9±2.8% in MHC II KO versus 39.0±4.2% in WT). MHC-II KO mice also displayed better microvascular perfusion than WT mice after 24hours of reperfusion. Also CD4 + T-cell sufficient OT-II mice, which express an in this context irrelevant T-cell receptor, revealed smaller infarct sizes compared to WT mice. However, MHC-II blocking anti-I-A/I-E antibody treatment was not able to reduce infarct size indicating that autoantigen recognition is not required for the activation of CD4 + T-cells during reperfusion. Flow-cytometric analysis also did not detect CD4 + T-cell activation in heart draining lymph nodes in response to 24hours of ischemia-reperfusion. Adoptive transfer of CD4 + T-cells in CD4 KO mice increased the infarct size only when including the Foxp3 + CD25 + subset. Depletion of CD4 + Foxp3 + T-cells in DEREG mice enabling specific conditional ablation of this subset by treatment with diphtheria toxin attenuated infarct size as compared to diphtheria toxin treated WT mice. CD4 + Foxp3 + T-cells enhance myocardial ischemia-reperfusion injury. CD4 + T-cells exert injurious effects without the need for prior activation by MHC-II restricted autoantigen recognition. Copyright © 2016 Elsevier Ltd. All rights reserved.

Improved fibronectin-immobilized fibrinogen microthreads for the attachment and proliferation of fibroblasts

PubMed Central

Rajangam, Thanavel; An, Seong Soo A

2013-01-01

The aim of this study was to fabricate fibrinogen (Fbg) microfibers with different structural characteristics for the development of 3-D tissue-engineering scaffolds. Fabricated Fbg microfibers were investigated for their biomolecule encapsulation, cell adhesion, and proliferations. Microfibers with three different concentrations of Fbg (5, 10, and 15 wt%) were prepared by a gel solvent-extraction method using a silicone rubber tube. Fbg microfibers were covalently modified with fibronectin (FN) by using water-soluble 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide as the cross-linking agent. Fbg microfibers were characterized by their FN cross-linking properties, structural morphology, and in vitro degradation. Furthermore, FN/Fbg microfibers were evaluated for cell attachment and proliferation. The bio-compatibility and cell proliferation of the microfibers were assessed by measuring adenosine triphosphate activity in C2C12 fibroblast cells. Cell attachment and proliferation on microfibers were further examined using fluorescence and scanning electron microscopic images. FN loading on the microfibers was confirmed by fluorescence and infrared spectroscopy. Surface morphology was characterized by scanning electron microscopy, and showed highly aligned nanostructures for fibers made with 15 wt% Fbg, a more porous structure for fibers made with 10 wt% Fbg, and a less porous structure for those made with 5 wt% Fbg. Controlled biodegradation of the fiber was observed for 8 weeks by using an in vitro proteolytic degradation assay. Fbg microfibers with highly aligned nanostructures (15 wt%) showed enhanced biomolecule encapsulation, as well as higher cell adhesion and proliferation than another two types of FN/Fbg fibers (5 and 10 wt%) and unmodified Fbg fibers. The promising results obtained from the present study reveal that optimal structure of Fbg microfibers could be used as a potential substratum for growth factors or drug release, especially in wound healing and vascular tissue engineering, in which fibers could be applied to promote and orient cell adhesion and proliferation. PMID:23515334

The Human 343delT HSPB5 Chaperone Associated with Early-onset Skeletal Myopathy Causes Defects in Protein Solubility*

PubMed Central

Mitzelfelt, Katie A.; Limphong, Pattraranee; Choi, Melinda J.; Kondrat, Frances D. L.; Lai, Shuping; Kolander, Kurt D.; Kwok, Wai-Meng; Dai, Qiang; Grzybowski, Michael N.; Zhang, Huali; Taylor, Graydon M.; Lui, Qiang; Thao, Mai T.; Hudson, Judith A.; Barresi, Rita; Bushby, Kate; Jungbluth, Heinz; Wraige, Elizabeth; Geurts, Aron M.; Benesch, Justin L. P.; Riedel, Michael; Christians, Elisabeth S.; Minella, Alex C.; Benjamin, Ivor J.

2016-01-01

Mutations of HSPB5 (also known as CRYAB or αB-crystallin), a bona fide heat shock protein and molecular chaperone encoded by the HSPB5 (crystallin, alpha B) gene, are linked to multisystem disorders featuring variable combinations of cataracts, cardiomyopathy, and skeletal myopathy. This study aimed to investigate the pathological mechanisms involved in an early-onset myofibrillar myopathy manifesting in a child harboring a homozygous recessive mutation in HSPB5, 343delT. To study HSPB5 343delT protein dynamics, we utilize model cell culture systems including induced pluripotent stem cells derived from the 343delT patient (343delT/343delT) along with isogenic, heterozygous, gene-corrected control cells (WT KI/343delT) and BHK21 cells, a cell line lacking endogenous HSPB5 expression. 343delT/343delT and WT KI/343delT-induced pluripotent stem cell-derived skeletal myotubes and cardiomyocytes did not express detectable levels of 343delT protein, contributable to the extreme insolubility of the mutant protein. Overexpression of HSPB5 343delT resulted in insoluble mutant protein aggregates and induction of a cellular stress response. Co-expression of 343delT with WT prevented visible aggregation of 343delT and improved its solubility. Additionally, in vitro refolding of 343delT in the presence of WT rescued its solubility. We demonstrate an interaction between WT and 343delT both in vitro and within cells. These data support a loss-of-function model for the myopathy observed in the patient because the insoluble mutant would be unavailable to perform normal functions of HSPB5, although additional gain-of-function effects of the mutant protein cannot be excluded. Additionally, our data highlight the solubilization of 343delT by WT, concordant with the recessive inheritance of the disease and absence of symptoms in carrier individuals. PMID:27226619

Smad4 in T cells plays a protective role in the development of autoimmune Sjögren's syndrome in the nonobese diabetic mouse.

PubMed

Kim, Donghee; Kim, Jae Young; Jun, Hee-Sook

2016-12-06

We investigated the role of Smad4, a signaling molecule of the TGF-beta pathway, in T cells on the pathology of Sjögren's syndrome (SS) in nonobese diabetic (NOD) mice, an animal model of SS. T cell-specific Smad4-deleted (Smad4fl/fl,CD4-Cre; Smad4 tKO) NOD mice had accelerated development of SS compared with wild-type (Smad4+/+,CD4-Cre; WT) NOD mice, including increased lymphocyte infiltration into exocrine glands, decreased tear and saliva production, and increased levels of autoantibodies at 12 weeks of age. Activated/memory T cells and cytokine (IFN-γ, IL-17)-producing T cells were increased in Smad4 tKO NOD mice, however the proportion and function of regulatory T (Treg) cells were not different between Smad4 tKO and WT NOD mice. Effector T (Teff) cells from Smad4 tKO NOD mice were less sensitive than WT Teff cells to suppression by Treg cells. Th17 differentiation capability of Teff cells was similar between Smad4 tKO and WT NOD mice, but IL-17 expression was increased under inducible Treg skewing conditions in T cells from Smad4 tKO NOD mice. Our results demonstrate that disruption of the Smad4 pathway in T cells of NOD mice increases Teff cell activation resulting in upregulation of Th17 cells, indicating that Smad4 in T cells has a protective role in the development of SS in NOD mice.

Smad4 in T cells plays a protective role in the development of autoimmune Sjögren's syndrome in the nonobese diabetic mouse

PubMed Central

Kim, Donghee; Kim, Jae Young; Jun, Hee-Sook

2016-01-01

We investigated the role of Smad4, a signaling molecule of the TGF-beta pathway, in T cells on the pathology of Sjögren's syndrome (SS) in nonobese diabetic (NOD) mice, an animal model of SS. T cell-specific Smad4-deleted (Smad4fl/fl,CD4-Cre; Smad4 tKO) NOD mice had accelerated development of SS compared with wild-type (Smad4+/+,CD4-Cre; WT) NOD mice, including increased lymphocyte infiltration into exocrine glands, decreased tear and saliva production, and increased levels of autoantibodies at 12 weeks of age. Activated/memory T cells and cytokine (IFN-γ, IL-17)-producing T cells were increased in Smad4 tKO NOD mice, however the proportion and function of regulatory T (Treg) cells were not different between Smad4 tKO and WT NOD mice. Effector T (Teff) cells from Smad4 tKO NOD mice were less sensitive than WT Teff cells to suppression by Treg cells. Th17 differentiation capability of Teff cells was similar between Smad4 tKO and WT NOD mice, but IL-17 expression was increased under inducible Treg skewing conditions in T cells from Smad4 tKO NOD mice. Our results demonstrate that disruption of the Smad4 pathway in T cells of NOD mice increases Teff cell activation resulting in upregulation of Th17 cells, indicating that Smad4 in T cells has a protective role in the development of SS in NOD mice. PMID:27880731

Transient development of ovotestes in XX Sox9 transgenic mice

PubMed Central

Gregoire, Elodie P.; Lavery, Rowena; Chassot, Anne-Amandine; Akiyama, Haruhiko; Treier, Mathias; Behringer, Richard R.; Chaboissier, Marie-Christine

2010-01-01

The sex of an individual results from the paternal transmission of the SRY gene located on the Y chromosome. In turn, SRY initiates Sox9 expression, a transcription factor required for testicular differentiation. Ectopic activation of SOX9 in XX Wt1:Sox9 transgenic mice, induces female-to-male sex reversal in adult mice. Here we show that complete sex reversal is preceded by a transient phase of ovotestis differentiation with XX Wt1:Sox9 transgenic gonads containing a testicular central region and one or both ovarian poles indicating that Wt1:Sox9 is not as efficient as Sry to induce male development. In XX Wt1:Sox9Tg/+ gonads, transgenic Sox9 is expressed earlier than Sox9 in XY gonads, and is able to induce the expression of EGFP, knocked into the 3′ UTR of Sox9 indicating that SOX9 is involved in the initiation and maintenance of its own expression. However, the delayed onset of expression of endogenous Sox9-EGFP suggests that this activation requires other factors, whose expression depends on SOX9. In the testicular regions of the XX Wt1:Sox9 ovotestes, proliferation of the XX foetal germ cells is hampered and they differentiate as pro-spermatogonia. This indicates that XX germ cells are not competent to respond to proliferative signals released from a testicular environment. In the ovarian regions, despite the continuous mRNA expression of the WT1:Sox9 transgene, the SOX9 protein does not accumulate suggesting that regulation of this gene in ovarian cells involves post-transcriptional mechanisms. Finally, ovarian cells of the XX Wt1:Sox9 ovotestis undergo apoptosis during late embryogenesis leading to complete female-to-male sex reversal of the transgenic mice at birth. PMID:20965161

Innate immune signaling by Toll-like receptor-4 (TLR4) shapes the inflammatory microenvironment in colitis-associated tumors

PubMed Central

Fukata, Masayuki; Hernandez, Yasmin; Conduah, Daisy; Cohen, Jason; Chen, Anli; Breglio, Keith; Goo, Tyralee; Hsu, David; Xu, Ruliang; Abreu, Maria T.

2009-01-01

Patients with ulcerative colitis are at increased risk for developing colorectal cancer. We have shown that TLR4 is over-expressed in human colitis-associated cancer (CAC) and that mice deficient in TLR4 are markedly protected against colitis-associated neoplasia. We wished to elucidate the specific contributions of TLR4 signaling by myeloid cells and colonic epithelial cells (CEC) in colitis-associated tumorigenesis. TLR4-deficient mice or wild-type littermates (WT) were transplanted with bone marrow (BM) cells: TLR4-/- BM→WT mice (TLR4-expressing CEC) and WT BM→TLR4-/- mice (TLR4-expressing myeloid cells). Colitis-associated neoplasia was induced by azoxymethane (AOM 7.3mg/kg) injection and two cycles of dextran sodium sulfate (DSS) treatment. The number and size of dysplastic lesions were greater in TLR4-/- BM→WT mice than in WT BM→TLR4-/- mice (P<0.005). Histologically, TLR4-/- BM→WT mice had greater numbers of mucosal neutrophils and macrophages compared to WT BM→TLR4-/- mice. The chemokines KC and CCL2, important in recruitment of neutrophils and macrophages, respectively, were induced in mice expressing TLR4 in CEC rather than the myeloid compartment. The lamina propria infiltrate of mice expressing TLR4 in CEC was characterized by macrophages expressing Cox-2. Moreover, mice expressing TLR4 in CEC rather than the myeloid compartment had increased production of amphiregulin and EGFR activation. These findings indicate that TLR4 signaling on CEC is necessary for recruitment and activation of Cox-2 expressing macrophages and increasing the number and size of dysplastic lesions. Our results implicate innate immune signaling on CEC as a key regulator of a tumor-promoting microenvironment. PMID:19229991

Degradable biocomposite of nano calcium-deficient hydroxyapatite-multi(amino acid) copolymer

PubMed Central

Li, Hong; Gong, Min; Yang, Aiping; Ma, Jian; Li, Xiangde; Yan, Yonggang

2012-01-01

Background and methods A nano calcium-deficient hydroxyapatite (n-CDHA)-multi(amino acid) copolymer (MAC) composite bone substitute biomaterial was prepared using an in situ polymerization method. The composition, structure, and compressive strength of the composite was characterized, and the in vitro degradability in phosphate-buffered solution and preliminary cell responses to the composite were investigated. Results The composite comprised n-CDHA and an amide linkage copolymer. The compressive strength of the composite was in the range of 88–129 MPa, varying with the amount of n-CDHA in the MAC (ranging from 10 wt% to 50 wt%). Weight loss from the composite increased (from 32.2 wt% to 44.3 wt%) with increasing n-CDHA content (from 10 wt% to 40 wt%) in the MAC after the composite was soaked in phosphate-buffered solution for 12 weeks. The pH of the soaking medium varied from 6.9 to 7.5. MG-63 cells with an osteogenic phenotype were well adhered and spread on the composite surface. Viability and differentiation increased with time, indicating that the composite had no negative effects on MG-63 cells. Conclusion The n-CDHA-MAC composite had good cytocompatibility and has potential to be used as a bone substitute. PMID:22457591

CD22 is required for formation of memory B cell precursors within germinal centers.

PubMed

Chappell, Craig P; Draves, Kevin E; Clark, Edward A

2017-01-01

CD22 is a BCR co-receptor that regulates B cell signaling, proliferation and survival and is required for T cell-independent Ab responses. To investigate the role of CD22 during T cell-dependent (TD) Ab responses and memory B cell formation, we analyzed Ag-specific B cell responses generated by wild-type (WT) or CD22-/- B cells following immunization with a TD Ag. CD22-/- B cells mounted normal early Ab responses yet failed to generate either memory B cells or long-lived plasma cells, whereas WT B cells formed both populations. Surprisingly, B cell expansion and germinal center (GC) differentiation were comparable between WT and CD22-/- B cells. CD22-/- B cells, however, were significantly less capable of generating a population of CXCR4hiCD38hi GC B cells, which we propose represent memory B cell precursors within GCs. These results demonstrate a novel role for CD22 during TD humoral responses evident during primary GC formation and underscore that CD22 functions not only during B cell maturation but also during responses to both TD and T cell-independent antigens.

CD22 is required for formation of memory B cell precursors within germinal centers

PubMed Central

Chappell, Craig P.; Draves, Kevin E.

2017-01-01

CD22 is a BCR co-receptor that regulates B cell signaling, proliferation and survival and is required for T cell-independent Ab responses. To investigate the role of CD22 during T cell-dependent (TD) Ab responses and memory B cell formation, we analyzed Ag-specific B cell responses generated by wild-type (WT) or CD22-/- B cells following immunization with a TD Ag. CD22-/- B cells mounted normal early Ab responses yet failed to generate either memory B cells or long-lived plasma cells, whereas WT B cells formed both populations. Surprisingly, B cell expansion and germinal center (GC) differentiation were comparable between WT and CD22-/- B cells. CD22-/- B cells, however, were significantly less capable of generating a population of CXCR4hiCD38hi GC B cells, which we propose represent memory B cell precursors within GCs. These results demonstrate a novel role for CD22 during TD humoral responses evident during primary GC formation and underscore that CD22 functions not only during B cell maturation but also during responses to both TD and T cell-independent antigens. PMID:28346517

B cells are critical to T-cell-mediated antitumor immunity induced by a combined immune-stimulatory/conditionally cytotoxic therapy for glioblastoma.

PubMed

Candolfi, Marianela; Curtin, James F; Yagiz, Kader; Assi, Hikmat; Wibowo, Mia K; Alzadeh, Gabrielle E; Foulad, David; Muhammad, A K M G; Salehi, Sofia; Keech, Naomi; Puntel, Mariana; Liu, Chunyan; Sanderson, Nicholas R; Kroeger, Kurt M; Dunn, Robert; Martins, Gislaine; Lowenstein, Pedro R; Castro, Maria G

2011-10-01

We have demonstrated that modifying the tumor microenvironment through intratumoral administration of adenoviral vectors (Ad) encoding the conditional cytotoxic molecule, i.e., HSV1-TK and the immune-stimulatory cytokine, i.e., fms-like tyrosine kinase 3 ligand (Flt3L) leads to T-cell-dependent tumor regression in rodent models of glioblastoma. We investigated the role of B cells during immune-mediated glioblastoma multiforme regression. Although treatment with Ad-TK+Ad-Flt3L induced tumor regression in 60% of wild-type (WT) mice, it completely failed in B-cell-deficient Igh6(-/-) mice. Tumor-specific T-cell precursors were detected in Ad-TK+Ad-Flt3L-treated WT mice but not in Igh6(-/-) mice. The treatment also failed in WT mice depleted of total B cells or marginal zone B cells. Because we could not detect circulating antibodies against tumor cells and the treatment was equally efficient in WT mice and in mice with B-cell-specific deletion of Prdm 1 (encoding Blimp-1), in which B cells are present but unable to fully differentiate into antibody-secreting plasma cells, tumor regression in this model is not dependent on B cells' production of tumor antigen-specific immunoglobulins. Instead, B cells seem to play a role as antigen-presenting cells (APCs). Treatment with Ad-TK+Ad-Flt3L led to an increase in the number of B cells in the cervical lymph nodes, which stimulated the proliferation of syngeneic T cells and induced clonal expansion of antitumor T cells. Our data show that B cells act as APCs, playing a critical role in clonal expansion of tumor antigen-specific T cells and brain tumor regression.

High Mobility Group Box 1 Promotes Angiogenesis from Bone Marrow-derived Endothelial Progenitor Cells after Myocardial Infarction.

PubMed

Nakamura, Yuichi; Suzuki, Satoshi; Shimizu, Takeshi; Miyata, Makiko; Shishido, Tetsuro; Ikeda, Kazuhiko; Saitoh, Shu-Ichi; Kubota, Isao; Takeishi, Yasuchika

2015-01-01

High mobility group box 1 (HMGB1) is a DNA-binding protein secreted into the extracellular space from necrotic cells that acts as a cytokine. We examined the role of HMGB1 in angiogenesis from bone marrow-derived cells in the heart using transgenic mice exhibiting the cardiac-specific overexpression of HMGB1 (HMGB1-TG). HMGB1-TG mice and wild-type littermate (WT) mice were lethally irradiated and injected with bone marrow cells from green fluorescent protein mice through the tail vein. After bone marrow transplantation, the left anterior descending artery was ligated to induce myocardial infarction (MI). Flow cytometry revealed that the levels of circulating endothelial progenitor cells (EPCs) mobilized from the bone marrow increased after MI in the HMGB-TG mice versus the WT mice. In addition, the size of MI was smaller in the HMGB1-TG mice than in the WT mice, and immunofluorescence staining demonstrated that the number of engrafted vascular endothelial cells derived from bone marrow in the border zones of the MI areas was increased in the HMGB1-TG mice compared to that observed in the WT mice. Moreover, the levels of cardiac vascular endothelial growth factor after MI were higher in the HMGB1-TG mice than in the WT mice. The present study demonstrated that HMGB1 promotes angiogenesis and reduces the MI size by enhancing the mobilization and differentiation of bone marrow cells to EPCs as well as their migration to the border zones of the MI areas and engraftment as vascular endothelial cells in new capillaries or arterioles in the infarcted heart.

Mesenchymal stem cells induce epithelial proliferation within the inflamed stomach.

PubMed

Donnelly, Jessica M; Engevik, Amy; Feng, Rui; Xiao, Chang; Boivin, Gregory P; Li, Jing; Houghton, JeanMarie; Zavros, Yana

2014-06-15

Bone marrow-derived mesenchymal stem cells (MSCs) sustain cancer cells by creating a microenvironment favorable for tumor growth. In particular, MSCs have been implicated in gastric cancer development. There is extensive evidence suggesting that Hedgehog signaling regulates tumor growth. However, very little is known regarding the precise roles of Hedgehog signaling and MSCs in tumor development within the stomach. The current study tests that hypothesis that Sonic Hedgehog (Shh), secreted from MSCs, provides a proliferative stimulus for the gastric epithelium in the presence of inflammation. Red fluorescent protein-expressing MSCs transformed in vitro (stMSCs) were transduced with lentiviral constructs containing a vector control (stMSC(vect)) or short hairpin RNA (shRNA) targeting the Shh gene (stMSC(ShhKO)). Gastric submucosal transplantation of wild-type MSCs (wtMSCs), wild-type MSCs overexpressing Shh (wtMSC(Shh)), stMSC(vect), or stMSC(ShhKO) cells in C57BL/6 control (BL/6) or gastrin-deficient (GKO) mice was performed and mice analyzed 30 and 60 days posttransplantation. Compared with BL/6 mice transplanted with wtMSC(Shh) and stMSC(vect) cells, inflamed GKO mice developed aggressive gastric tumors. Tumor development was not observed in mouse stomachs transplanted with wtMSC or stMSC(ShhKO) cells. Compared with stMSC(ShhKO)-transplanted mice, within the inflamed GKO mouse stomach, Shh-expressing stMSC(vect)- and wtMSC(Shh)-induced proliferation of CD44-positive cells. CD44-positive cells clustered in gland-like structures within the tumor stroma and were positive for Patched (Ptch) expression. We conclude that Shh, secreted from MSCs, provides a proliferative stimulus for the gastric epithelium that is associated with tumor development, a response that is sustained by chronic inflammation. Copyright © 2014 the American Physiological Society.

Mesenchymal stem cells induce epithelial proliferation within the inflamed stomach

PubMed Central

Donnelly, Jessica M.; Engevik, Amy; Feng, Rui; Xiao, Chang; Boivin, Gregory P.; Li, Jing; Houghton, JeanMarie

2014-01-01

Bone marrow-derived mesenchymal stem cells (MSCs) sustain cancer cells by creating a microenvironment favorable for tumor growth. In particular, MSCs have been implicated in gastric cancer development. There is extensive evidence suggesting that Hedgehog signaling regulates tumor growth. However, very little is known regarding the precise roles of Hedgehog signaling and MSCs in tumor development within the stomach. The current study tests that hypothesis that Sonic Hedgehog (Shh), secreted from MSCs, provides a proliferative stimulus for the gastric epithelium in the presence of inflammation. Red fluorescent protein-expressing MSCs transformed in vitro (stMSCs) were transduced with lentiviral constructs containing a vector control (stMSCvect) or short hairpin RNA (shRNA) targeting the Shh gene (stMSCShhKO). Gastric submucosal transplantation of wild-type MSCs (wtMSCs), wild-type MSCs overexpressing Shh (wtMSCShh), stMSCvect, or stMSCShhKO cells in C57BL/6 control (BL/6) or gastrin-deficient (GKO) mice was performed and mice analyzed 30 and 60 days posttransplantation. Compared with BL/6 mice transplanted with wtMSCShh and stMSCvect cells, inflamed GKO mice developed aggressive gastric tumors. Tumor development was not observed in mouse stomachs transplanted with wtMSC or stMSCShhKO cells. Compared with stMSCShhKO-transplanted mice, within the inflamed GKO mouse stomach, Shh-expressing stMSCvect- and wtMSCShh-induced proliferation of CD44-positive cells. CD44-positive cells clustered in gland-like structures within the tumor stroma and were positive for Patched (Ptch) expression. We conclude that Shh, secreted from MSCs, provides a proliferative stimulus for the gastric epithelium that is associated with tumor development, a response that is sustained by chronic inflammation. PMID:24789207

The Effect of PKCα on the Light Response of Rod Bipolar Cells in the Mouse Retina

PubMed Central

Xiong, Wei-Hong; Pang, Ji-Jie; Pennesi, Mark E.; Duvoisin, Robert M.; Wu, Samuel M.; Morgans, Catherine W.

2015-01-01

Purpose Protein kinase C α (PKCα) is abundantly expressed in rod bipolar cells (RBCs) in the retina, yet the physiological function of PKCα in these cells is not well understood. To elucidate the role of PKCα in visual processing in the eye, we examined the effect of genetic deletion of PKCα on the ERG and on RBC light responses in the mouse. Methods Immunofluorescent labeling was performed on wild-type (WT), TRPM1 knockout, and PKCα knockout (PKC-KO) retina. Scotopic and photopic ERGs were recorded from WT and PKC-KO mice. Light responses of RBCs were measured using whole-cell recordings in retinal slices from WT and PKC-KO mice. Results Protein kinase C alpha expression in RBCs is correlated with the activity state of the cell. Rod bipolar cells dendrites are a major site of PKCα phosphorylation. Electroretinogram recordings indicated that loss of PKCα affects the scotopic b-wave, including a larger peak amplitude, longer implicit time, and broader width of the b-wave. There were no differences in the ERG a- or c-wave between PKCα KO and WT mice, indicating no measurable effect of PKCα in photoreceptors or the RPE. The photopic ERG was unaffected consistent with the lack of detectable PKCα in cone bipolar cells. Whole-cell recordings from RBCs in PKC-KO retinal slices revealed that, compared with WT, RBC light responses in the PKC-KO retina are delayed and of longer duration. Conclusions Protein kinase C alpha plays an important modulatory role in RBCs, regulating both the peak amplitude and temporal properties of the RBC light response in the rod visual pathway. PMID:26230760

Disease-associated mutations in CNGB3 promote cytotoxicity in photoreceptor-derived cells

PubMed Central

Liu, Chunming; Sherpa, Tshering

2013-01-01

Purpose To determine if achromatopsia associated F525N and T383fsX mutations in the CNGB3 subunit of cone photoreceptor cyclic nucleotide-gated (CNG) channels increases susceptibility to cell death in photoreceptor-derived cells. Methods Photoreceptor-derived 661W cells were transfected with cDNA encoding wild-type (WT) CNGA3 subunits plus WT or mutant CNGB3 subunits, and incubated with the membrane-permeable CNG channel activators 8-(4-chlorophenylthio) guanosine 3′,5′-cyclic monophosphate (CPT-cGMP) or CPT-adenosine 3′,5′-cyclic monophosphate (CPT-cAMP). Cell viability under these conditions was determined by measuring lactate dehydrogenase release. Channel ligand sensitivity was calibrated by patch-clamp recording after expression of WT or mutant channels in Xenopus oocytes. Results Coexpression of CNGA3 with CNGB3 subunits containing F525N or T383fsX mutations produced channels exhibiting increased apparent affinity for CPT-cGMP compared to WT channels. Consistent with these effects, cytotoxicity in the presence of 0.1 μM CPT-cGMP was enhanced relative to WT channels, and the increase in cell death was more pronounced for the mutation with the largest gain-of-function effect on channel gating, F525N. Increased susceptibility to cell death was prevented by application of the CNG channel blocker L-cis-diltiazem. Increased cytotoxicity was also found to be dependent on the presence of extracellular calcium. Conclusions These results indicate a connection between disease-associated mutations in cone CNG channel subunits, altered CNG channel-activation properties, and photoreceptor cytotoxicity. The rescue of cell viability via CNG channel block or removal of extracellular calcium suggests that cytotoxicity in this model depends on calcium entry through hyperactive CNG channels. PMID:23805033

Physiologic TLR9-CpG-DNA interaction is essential for the homeostasis of the intestinal immune system.

PubMed

Hofmann, Claudia; Dunger, Nadja; Doser, Kristina; Lippert, Elisabeth; Siller, Sebastian; Edinger, Matthias; Falk, Werner; Obermeier, Florian

2014-01-01

Cytosine-guanosine dinucleotide (CpG) motifs are immunostimulatory components of bacterial DNA and activators of innate immunity through Toll-like receptor 9 (TLR9). Administration of CpG oligodeoxynucleotides before the onset of experimental colitis prevents intestinal inflammation by enforcement of regulatory mechanisms. It was investigated whether physiologic CpG/TLR9 interactions are critical for the homeostasis of the intestinal immune system. Mesenteric lymph node cell and lamina propria mononuclear cell (LPMC) populations from BALB/c wild-type (wt) or TLR9 mice were assessed by flow cytometry and proteome profiling. Cytokine secretion was determined and nuclear extracts were analyzed for nuclear factor kappa B (NF-κB) and cAMP response-element binding protein activity. To assess the colitogenic potential of intestinal T cells, CD4-enriched cells from LPMC of wt or TLR9 donor mice were injected intraperitoneally in recipient CB-17 SCID mice. TLR9 deficiency was accompanied by slight changes in cellular composition and phosphorylation of signaling proteins of mesenteric lymph node cell and LPMC. LPMC from TLR9 mice displayed an increased proinflammatory phenotype compared with wt LPMC. NF-κB activity in cells from TLR9 mice was enhanced, whereas cAMP response-element binding activity was reduced compared with wt. Transfer of lamina propria CD4-enriched T cells from TLR9 mice induced severe colitis, whereas wt lamina propria CD4-enriched T cells displayed an attenuated phenotype. Lack of physiologic CpG/TLR9 interaction impairs the function of the intestinal immune system indicated by enhanced proinflammatory properties. Thus, physiologic CpG/TLR interaction is essential for homeostasis of the intestinal immune system as it is required for the induction of counterregulating anti-inflammatory mechanisms.

Effect of Drying Methods on Protein and DNA Conformation Changes in Lactobacillus rhamnosus GG Cells by Fourier Transform Infrared Spectroscopy.

PubMed

Hlaing, Mya M; Wood, Bayden R; McNaughton, Don; Ying, DanYang; Dumsday, Geoff; Augustin, Mary Ann

2017-03-01

Microencapsulation protects cells against environmental stress encountered during the production of probiotics, which are used as live microbial food ingredients. Freeze-drying and spray-drying are used in the preparation of powdered microencapsulated probiotics. This study examines the ability of Fourier transform infrared (FTIR) spectroscopy to detect differences in cells exposed to freeze-drying and spray-drying of encapsulated Lactobacillus rhamnosus GG cells. The FTIR analysis clearly demonstrated there were more significant molecular changes in lipid, fatty acid content, protein, and DNA conformation of nonencapsulated compared to encapsulated bacterial cells. The technique was also able to differentiate between spray-dried and freeze-dried cells. The results also revealed the extent of protection from a protein-carbohydrate-based encapsulant matrix on the cells depending on the type drying process. The extent of this protection to the dehydration stress was shown to be less in spray-dried cells than in freeze-dried cells. This suggests that FTIR could be used as a rapid, noninvasive, and real-time measurement technique to detect detrimental drying effects on cells.

T-type Ca2+ channels regulate the exit of cardiac myocytes from the cell cycle after birth

PubMed Central

Wang, Fang; Gao, Hui; Kubo, Hajime; Fan, Xiaoxuan; Zhang, Hongyu; Berretta, Remus; Chen, Xiongwen; Sharp, Thomas; Starosta, Timothy; Makarewich, Catherine; Li, Ying; Molkentin, Jeffrey D.; Houser, Steven R.

2013-01-01

T-type Ca2+ channels (TTCCs) are expressed in the fetal heart and then disappear from ventricular myocytes after birth. The hypothesis examined in this study was the α1G TTCCs' influence in myocyte maturation and their rapid withdrawal from the cell cycle after birth. Methods Cardiac myocytes were isolated from neonatal and adult wild type (WT), α1G−/− and α1G over expressing (α1GDT) mice. Bromodeoxyuridine (BrdU) uptake, myocyte nucleation, cell cycle analysis, and T-type Ca2+ currents were measured. Results All myocytes were mono-nucleated at birth and 35% of WT myocytes expressed functional TTCCs. Very few neonatal myocytes had functional TTCCs in α1G−/− hearts. By the end of the first week after birth no WT or α1G−/− had functional TTCCs. During the first week after birth about 25% of WT myocytes were BrdU+ and became bi-nucleated. Significantly fewer α1G−/− myocytes became bi-nucleated and fewer of these myocytes were BrdU+. Neonatal α1G−/− myocytes were also smaller than WT. Adult WT and α1G−/− hearts were similar in size, but α1G−/− myocytes were smaller and a greater % were mono-nucleated. α1G over expressing hearts were smaller than WT but their myocytes were larger. Conclusions The studies performed show that loss of functional TTCCs is associated with bi-nucleation and myocyte withdrawal from the cell cycle. Loss of α1G TTCCs slowed the transition from mono- to bi-nucleation and resulted in an adult heart with a greater number of small cardiac myocytes. These results suggest that TTCCs are involved in the regulation of myocyte size and the exit of myocytes from the cell cycle during the first week after birth. PMID:23743021

DOE Office of Scientific and Technical Information (OSTI.GOV)

Adhikari, Ananta R., E-mail: aa8381@gmail.com; Rusakova, Irene; Chu, Wei-Kan

Polymer-matrix nanocomposites based on Poly(lactic-co-glycolic) acid (PLGA) and Graphene platelets (GNPs) were studied. GNPs, nanomaterials with a 2D flat surface, were chosen with or without chemical modification in PLGA/GNP nanocomposites and their microstructure, thermal property, and their compatibility as scaffolds for cell growth were investigated. PLGA/GNP nanocomposites (0, 1, and 5 wt. % of GNPs) were prepared using a solution based technique. Transmission electron microscopy, X-ray diffraction, Differential scanning calorimeter, and Thermogravimetric analyzer were used to analyze morphology and thermal properties. This work demonstrated the role of GNPs flat surface to provide a favorable platform resulting in an enhanced PLGA crystallization. Functionalizedmore » GNPs suppress both the thermal stability and the crystallization of PLGA. Finally, to determine the potential usefulness of these scaffolds for biomedical applications, mammalian cells were cultured on various PLGA/GNP nanocomposites (0, 1, and 5 wt. % GNPs). 1 wt. % PLGA/GNP nanocomposites showed better biocompatibility for cell growth with/without graphenes functionalization compared to pure PLGA and 5 wt. % PLGA/GNP. The function of GNPs in PLGA/GNPs (1 wt. %) composites is to provide a stage for PLGA crystallization where cell growth is favored. These results provide strong evidence for a new class of materials that could be important for biomedical applications.« less

Capillary arterialization requires the bone-marrow-derived cell (BMC)-specific expression of chemokine (C-C motif) receptor-2, but BMCs do not transdifferentiate into microvascular smooth muscle.

PubMed

Nickerson, Meghan M; Burke, Caitlin W; Meisner, Joshua K; Shuptrine, Casey W; Song, Ji; Price, Richard J

2009-01-01

Chemokine (C-C motif) receptor-2 (CCR2) regulates arteriogenesis and angiogenesis, facilitating the MCP-1-dependent recruitment of growth factor-secreting bone marrow-derived cells (BMCs). Here, we tested the hypothesis that the BMC-specific expression of CCR2 is also required for new arteriole formation via capillary arterialization. Following non-ischemic saphenous artery occlusion, we measured the following in gracilis muscles: monocyte chemotactic protein-1 (MCP-1) in wild-type (WT) C57Bl/6J mice by ELISA, and capillary arterialization in WT-WT and CCR2(-/-)-WT (donor-host) bone marrow chimeric mice, as well as BMC transdifferentiation in EGFP(+)-WT mice, by smooth muscle (SM) alpha-actin immunochemistry. MCP-1 levels were significantly elevated 1 day after occlusion in WT mice. In WT-WT mice at day 7, compared to sham controls, arterial occlusion induced a 34% increase in arteriole length density, a 46% increase in SM alpha-actin(+) vessels, and a 45% increase in the fraction of vessels coated with SM alpha-actin, indicating significant capillary arterialization. However, in CCR2(-/-)-WT mice, no differences were observed between arterial occlusion and sham surgery. In EGFP(+)-WT mice, EGFP and SM alpha-actin never colocalized. We conclude that BMC-specific CCR2 expression is required for skeletal muscle capillary arterialization following arterial occlusion; however, BMCs do not transdifferentiate into smooth muscle.

Er0.4Bi1.6O3-δ - La0.8Sr0.2MnO3-δ nano-composite as a low-temperature firing cathode of solid oxide fuel cell

NASA Astrophysics Data System (ADS)

Kim, Sun Jae; Dayaghi, Amir Masoud; Kim, Kun Joong; Choi, Gyeong Man

2017-03-01

Er0.4Bi1.6O3-δ (ESB) composited with La0.8Sr0.2MnO3-δ (LSM) (2:3 or 3:2 wt:wt) with a bonding aid to decrease firing temperature TF are screen-printed on symmetric single cells composed of a Gd0.2Ce0.8O2-δ (GDC) interlayer/yttria-stabilized zirconia (YSZ) electrolyte/GDC interlayer, and their impedance spectra are compared. Addition of 5 wt % CuO to ESB-LSM (3:2 wt:wt) decreases the cathode TF to 650 °C without increasing cathodic polarization resistance (Rp ∼0.19 Ω cm2 at 650 °C). This ESB-LSM composite can be used as a cathode that can be fired at low temperature.

Impaired Angiogenesis and Mobilization of Circulating Angiogenic Cells in HIF-1α Heterozygous-Null Mice after Burn Wounding

PubMed Central

Zhang, Xianjie; Liu, Lixin; Wei, Xiaofei; Tan, Yee Sun; Tong, Lana; Chang, Ryan; Ghanamah, Mohammed S.; Reinblatt, Maura; Marti, Guy P.; Harmon, John W.; Semenza, Gregg L.

2014-01-01

Hypoxia-inducible factor 1 (HIF-1) is a transcription factor that controls vascular responses to hypoxia and ischemia. In this study, mice that were heterozygous for a null allele at the locus encoding the HIF-1α subunit (HET mice) and their wild type (WT) littermates were subjected to thermal injury involving 10% of body surface area. HIF-1α protein levels were increased in burn wounds of WT but not of HET mice on day 2. Serum levels of stromal-derived factor 1α, which binds to CXCR4, were increased on day 2 in WT but not in HET mice. Circulating angiogenic cells were also increased on day 2 in WT but not in HET mice and included CXCR4+Sca1+ cells. Laser Doppler perfusion imaging demonstrated increased blood flow in burn wounds of WT but not HET mice on day 7. Immunohistochemistry on day 7 revealed a reduced number of CD31+ vessels at the healing margin of burn wounds in HET as compared to WT mice. Vessel maturation was also impaired in wounds of HET mice as determined by the number of α-smooth muscle actin-positive vessels on day 21. The remaining wound area on day 14 was significantly increased in HET mice compared to WT littermates. The percentage of healed wounds on day 14 was significantly decreased in HET mice. These data delineate a signaling pathway by which HIF-1 promotes angiogenesis during burn wound healing. PMID:20163569

The effects of biochar and manure in silage corn

USDA-ARS?s Scientific Manuscript database

Amending soil with biochar may be a means of sequestering atmospheric CO2 and improving soil quality, but few multiyear field studies have examined the impacts of a one-time biochar application in an irrigated, calcareous soil. We fall-applied four treatments: dairy manure (18.7 tons/ac dry wt.); ha...

Activation of murine invariant NKT cells promotes susceptibility to candidiasis by IL-10 induced modulation of phagocyte antifungal activity.

PubMed

Haraguchi, Norihiro; Kikuchi, Norihiro; Morishima, Yuko; Matsuyama, Masashi; Sakurai, Hirofumi; Shibuya, Akira; Shibuya, Kazuko; Taniguchi, Masaru; Ishii, Yukio

2016-07-01

Invariant NKT (iNKT) cells play an important role in a variety of antimicrobial immune responses due to their ability to produce high levels of immune-modulating cytokines. Here, we investigated the role of iNKT cells in host defense against candidiasis using Jα18-deficient mice (Jα18(-/-) ), which lack iNKT cells. Jα18(-/-) mice were more resistant to the development of lethal candidiasis than wild-type (WT) mice. In contrast, treatment of WT mice with the iNKT cell activating ligand α-galactosylceramide markedly enhanced their mortality after infection with Candida albicans. Serum IL-10 levels were significantly elevated in WT mice in response to infection with C. albicans. Futhermore, IL-10 production increased after in vitro coculture of peritoneal macrophages with iNKT cells and C. albicans. The numbers of peritoneal macrophages, the production of IL-1β and IL-18, and caspase-1 activity were also significantly elevated in Jα18(-/-) mice after infection with C. albicans. The adoptive transfer of iNKT cells or exogenous administration of IL-10 into Jα18(-/-) reversed susceptibility to candidiasis to the level of WT mice. These results suggest that activation of iNKT cells increases the initial severity of C. albicans infection, most likely mediated by IL-10 induced modulation of macrophage antifungal activity. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Dilute phosphoric acid-catalysed hydrolysis of municipal bio-waste wood shavings using autoclave parr reactor system.

PubMed

Orozco, Angela M; Al-Muhtaseb, Ala'a H; Albadarin, Ahmad B; Rooney, David; Walker, Gavin M; Ahmad, Mohammad N M

2011-10-01

The visibility of using municipal bio-waste, wood shavings, as a potential feedstock for ethanol production was investigated. Dilute acid hydrolysis of wood shavings with H₃PO₄ was undertaken in autoclave parr reactor. A combined severity factor (CSF) was used to integrate the effects of hydrolysis times, temperature and acid concentration into a single variable. Xylose concentration reached a maximum value of 17 g/100 g dry mass corresponding to a yield of 100% at the best identified conditions of 2.5 wt.% H₃PO₄, 175 °C and 10 min reaction time corresponding to a CSF of 1.9. However, for glucose, an average yield of 30% was obtained at 5 wt.% H₃PO₄, 200 °C and 10 min. Xylose production increased with increasing temperature and acid concentration, but its transformation to the degradation product furfural was also catalysed by those factors. The maximum furfural formed was 3 g/100 g dry mass, corresponding to the 24% yield. Copyright © 2011 Elsevier Ltd. All rights reserved.

Glucose-ABL1-TOR Signaling Modulates Cell Cycle Tuning to Control Terminal Appressorial Cell Differentiation

PubMed Central

2017-01-01

The conserved target of rapamycin (TOR) pathway integrates growth and development with available nutrients, but how cellular glucose controls TOR function and signaling is poorly understood. Here, we provide functional evidence from the devastating rice blast fungus Magnaporthe oryzae that glucose can mediate TOR activity via the product of a novel carbon-responsive gene, ABL1, in order to tune cell cycle progression during infection-related development. Under nutrient-free conditions, wild type (WT) M. oryzae strains form terminal plant-infecting cells (appressoria) at the tips of germ tubes emerging from three-celled spores (conidia). WT appressorial development is accompanied by one round of mitosis followed by autophagic cell death of the conidium. In contrast, Δabl1 mutant strains undergo multiple rounds of accelerated mitosis in elongated germ tubes, produce few appressoria, and are abolished for autophagy. Treating WT spores with glucose or 2-deoxyglucose phenocopied Δabl1. Inactivating TOR in Δabl1 mutants or glucose-treated WT strains restored appressorium formation by promoting mitotic arrest at G1/G0 via an appressorium- and autophagy-inducing cell cycle delay at G2/M. Collectively, this work uncovers a novel glucose-ABL1-TOR signaling axis and shows it engages two metabolic checkpoints in order to modulate cell cycle tuning and mediate terminal appressorial cell differentiation. We thus provide new molecular insights into TOR regulation and cell development in response to glucose. PMID:28072818

Glucose-ABL1-TOR Signaling Modulates Cell Cycle Tuning to Control Terminal Appressorial Cell Differentiation.

PubMed

Marroquin-Guzman, Margarita; Sun, Guangchao; Wilson, Richard A

2017-01-01

The conserved target of rapamycin (TOR) pathway integrates growth and development with available nutrients, but how cellular glucose controls TOR function and signaling is poorly understood. Here, we provide functional evidence from the devastating rice blast fungus Magnaporthe oryzae that glucose can mediate TOR activity via the product of a novel carbon-responsive gene, ABL1, in order to tune cell cycle progression during infection-related development. Under nutrient-free conditions, wild type (WT) M. oryzae strains form terminal plant-infecting cells (appressoria) at the tips of germ tubes emerging from three-celled spores (conidia). WT appressorial development is accompanied by one round of mitosis followed by autophagic cell death of the conidium. In contrast, Δabl1 mutant strains undergo multiple rounds of accelerated mitosis in elongated germ tubes, produce few appressoria, and are abolished for autophagy. Treating WT spores with glucose or 2-deoxyglucose phenocopied Δabl1. Inactivating TOR in Δabl1 mutants or glucose-treated WT strains restored appressorium formation by promoting mitotic arrest at G1/G0 via an appressorium- and autophagy-inducing cell cycle delay at G2/M. Collectively, this work uncovers a novel glucose-ABL1-TOR signaling axis and shows it engages two metabolic checkpoints in order to modulate cell cycle tuning and mediate terminal appressorial cell differentiation. We thus provide new molecular insights into TOR regulation and cell development in response to glucose.

Disulfide bond exchanges in integrins αIIbβ3 and αvβ3 are required for activation and post-ligation signaling during clot retraction.

PubMed

Mor-Cohen, Ronit; Rosenberg, Nurit; Averbukh, Yulia; Seligsohn, Uri; Lahav, Judith

2014-05-01

Integrin αIIbβ3 mediates platelet adhesion, aggregation and fibrin clot retraction. These processes require activation of αIIbβ3 and post-ligation signaling. Disulfide bond exchanges are involved in αIIbβ3 and αvβ3 activation. In order to investigate the role of integrin activation and disulfide bond exchange during αIIbβ3- and αvβ3-mediated clot retraction, we co-expressed in baby hamster kidney cells wild-type (WT) human αIIb and WT or mutated human β3 that contain single or double cysteine substitutions disrupting C523-C544 or C560-C583 bonds. Flow cytometry was used to measure surface expression and activation state of the integrins. Time-course of fibrin clot retraction was examined. Cells expressed WT or mutated human αIIbβ3 as well as chimeric hamster/human αvβ3. The αIIbβ3 mutants were constitutively active and the thiol blocker dithiobisnitrobenzoic acid (DTNB) did not affect their activation state. WT cells retracted the clot and addition of αvβ3 inhibitors decreased the retraction rate. The active mutants and WT cells activated by anti-LIBS6 antibody retracted the clot faster than untreated WT cells, particularly in the presence of αvβ3 inhibitor. DTNB substantially inhibited clot retraction by WT or double C523S/C544S mutant expressing cells, but minimally affected single C523S, C544S or C560S mutants. Anti-LIBS6-enhanced clot retraction was significantly inhibited by DTNB when added prior to anti-LIBS6. Both αIIbβ3 and αvβ3 contribute to clot retraction without prior activation of the integrins. Activation of αIIbβ3, but not of αvβ3 enhances clot retraction. Both αIIbβ3 activation and post-ligation signaling during clot retraction require disulfide bond exchange. Copyright © 2014 Elsevier Ltd. All rights reserved.

Micromodel observations of evaporative drying and salt deposition in porous media

NASA Astrophysics Data System (ADS)

Rufai, Ayorinde; Crawshaw, John

2017-12-01

Most evaporation experiments using artificial porous media have focused on single capillaries or sand packs. We have carried out, for the first time, evaporation studies on a 2.5D micromodel based on a thin section of a sucrosic dolomite rock. This allowed direct visual observation of pore-scale processes in a network of pores. NaCl solutions from 0 wt. % (de-ionized water) to 36 wt. % (saturated brine) were evaporated by passing dry air through a channel in front of the micromodel matrix. For de-ionized water, we observed the three classical periods of evaporation: the constant rate period (CRP) in which liquid remains connected to the matrix surface, the falling rate period, and the receding front period, in which the capillary connection is broken and water transport becomes dominated by vapour diffusion. However, when brine was dried in the micromodel, we observed that the length of the CRP decreased with increasing brine concentration and became almost non-existent for the saturated brine. In the experiments with brine, the mass lost by evaporation became linear with the square root of time after the short CRP. However, this is unlikely to be due to capillary disconnection from the surface of the matrix, as salt crystals continued to be deposited in the channel above the matrix. We propose that this is due to salt deposition at the matrix surface progressively impeding hydraulic connectivity to the evaporating surface.

Irradiation effect of low-energy ion on polyurethane nanocoating containing metal oxide nanoparticles

NASA Astrophysics Data System (ADS)

Verma, Jaya; Nigam, Subhasha; Sinha, Surbhi; Sikarwar, B. S.; Bhattacharya, Arpita

2017-12-01

Irradiation effect of low-energy ion beam has been investigated on nanocoating developed with silica, titania and silica-titania core-shell nanoparticles embedded in an organic binder for nanopaint application. In this work, we have taken polyurethane as a model organic binder. Silica nanoparticles have been prepared through sol-gel synthesis with a particle size of 85 nm. Titania and core-shell nanoparticles have been prepared through both sol-gel and peptization process. Particle sizes obtained were 107 nm for titania and 240 nm for core-shell nanoparticles prepared through sol-gel process and 75 nm for TiO2 and 144 nm for core-shell nanoparticles prepared through peptization process. The coating formulations were developed with the above nanoparticles individually and nanoparticle concentration was varied from 1 to 6 wt% and the best performance in terms of hydrophobicity was obtained with 4 wt % of the nanoparticles in polyurethane coating formulation. All the coating formulations prepared were applied on a glass substrate and dried at 100°C. The dry film thickness obtained was around 100 µm in each case. These films dried on glass substrate were irradiated by nitrogen and argon ion beam with energy of 26 keV at fluences of 1014 to 1016 ions/cm2. The anti-algal property of the irradiated samples was improved and hydrophobicity was reduced.

TNFR2-deficient memory CD8 T cells provide superior protection against tumor cell growth.

PubMed

Kim, Edward Y; Teh, Soo-Jeet; Yang, Jocelyn; Chow, Michael T; Teh, Hung-Sia

2009-11-15

TNF receptor-2 (TNFR2) plays a critical role in promoting the activation and survival of naive T cells during the primary response. Interestingly, anti-CD3 plus IL-2 activated TNFR2(-/-) CD8 T cells are highly resistant to activation-induced cell death (AICD), which correlates with high expression levels of prosurvival molecules such as Bcl-2, survivin, and CD127 (IL-7Ralpha). We determined whether the resistance of activated TNFR2(-/-) CD8 T cells to AICD contributes to more effective protection against tumor cell growth. We found that during a primary tumor challenge, despite initial inferiority in controlling tumor cell growth, TNFR2(-/-) mice were able to more effectively control tumor burden over time compared with wild-type (WT) mice. Furthermore, vaccination of TNFR2(-/-) mice with recombinant Listeria monocytogenes that express OVA confers better protection against the growth of OVA-expressing E.G7 tumor cells relative to similarly vaccinated WT mice. The enhanced protection against tumor cell growth was not due to more effective activation of OVA-specific memory CD8 T cells in vaccinated TNFR2(-/-) mice. In vitro studies indicate that optimally activated OVA-specific TNFR2(-/-) CD8 T cells proliferated to the same extent and possess similar cytotoxicity against E.G7 tumor cells as WT CD8 T cells. However, relative to WT cells, activated OVA-specific TNFR2(-/-) CD8 T cells were highly resistant to AICD. Thus, the enhanced protection against E.G7 in TNFR2(-/-) mice is likely due to the recruitment and activation of OVA-specific memory TNFR2(-/-) CD8 T cells and their prolonged survival at the tumor site.

Cross-activating invariant NKT cells and kupffer cells suppress cholestatic liver injury in a mouse model of biliary obstruction.

PubMed

Duwaerts, Caroline C; Sun, Eric P; Cheng, Chao-Wen; van Rooijen, Nico; Gregory, Stephen H

2013-01-01

Both Kupffer cells and invariant natural killer T (iNKT) cells suppress neutrophil-dependent liver injury in a mouse model of biliary obstruction. We hypothesize that these roles are interdependent and require iNKT cell-Kupffer cell cross-activation. Female, wild-type and iNKT cell-deficient C57Bl/6 mice were injected with magnetic beads 3 days prior to bile duct ligation (BDL) in order to facilitate subsequent Kupffer cell isolation. On day three post-BDL, the animals were euthanized and the livers dissected. Necrosis was scored; Kupffer cells were isolated and cell surface marker expression (flow cytometry), mRNA expression (qtPCR), nitric oxide (NO (.) ) production (Griess reaction), and protein secretion (cytometric bead-array or ELISAs) were determined. To address the potential role of NO (.) in suppressing neutrophil accumulation, a group of WT mice received 1400W, a specific inducible nitric oxide synthase (iNOS) inhibitor, prior to BDL. To clarify the mechanisms underlying Kupffer cell-iNKT cell cross-activation, WT animals were administered anti-IFN-γ or anti-lymphocyte function-associated antigen (LFA)-1 antibody prior to BDL. Compared to their WT counterparts, Kupffer cells obtained from BDL iNKT cell-deficient mice expressed lower iNOS mRNA levels, produced less NO (.) , and secreted more neutrophil chemoattractants. Both iNOS inhibition and IFN-γ neutralization increased neutrophil accumulation in the livers of BDL WT mice. Anti-LFA-1 pre-treatment reduced iNKT cell accumulation in these same animals. These data indicate that the LFA-1-dependent cross-activation of iNKT cells and Kupffer cells inhibits neutrophil accumulation and cholestatic liver injury.

Cross-Activating Invariant NKT Cells and Kupffer Cells Suppress Cholestatic Liver Injury in a Mouse Model of Biliary Obstruction

PubMed Central

Duwaerts, Caroline C.; Sun, Eric P.; Cheng, Chao-Wen; van Rooijen, Nico; Gregory, Stephen H.

2013-01-01

Both Kupffer cells and invariant natural killer T (iNKT) cells suppress neutrophil-dependent liver injury in a mouse model of biliary obstruction. We hypothesize that these roles are interdependent and require iNKT cell-Kupffer cell cross-activation. Female, wild-type and iNKT cell-deficient C57Bl/6 mice were injected with magnetic beads 3 days prior to bile duct ligation (BDL) in order to facilitate subsequent Kupffer cell isolation. On day three post-BDL, the animals were euthanized and the livers dissected. Necrosis was scored; Kupffer cells were isolated and cell surface marker expression (flow cytometry), mRNA expression (qtPCR), nitric oxide (NO.) production (Griess reaction), and protein secretion (cytometric bead-array or ELISAs) were determined. To address the potential role of NO. in suppressing neutrophil accumulation, a group of WT mice received 1400W, a specific inducible nitric oxide synthase (iNOS) inhibitor, prior to BDL. To clarify the mechanisms underlying Kupffer cell-iNKT cell cross-activation, WT animals were administered anti-IFN-γ or anti-lymphocyte function-associated antigen (LFA)-1 antibody prior to BDL. Compared to their WT counterparts, Kupffer cells obtained from BDL iNKT cell-deficient mice expressed lower iNOS mRNA levels, produced less NO., and secreted more neutrophil chemoattractants. Both iNOS inhibition and IFN-γ neutralization increased neutrophil accumulation in the livers of BDL WT mice. Anti-LFA-1 pre-treatment reduced iNKT cell accumulation in these same animals. These data indicate that the LFA-1-dependent cross-activation of iNKT cells and Kupffer cells inhibits neutrophil accumulation and cholestatic liver injury. PMID:24260285

Wilms' Tumor 1 Overexpression in Granulosa Cells Is Associated with Polycystic Ovaries in Polycystic Ovary Syndrome Patients.

PubMed

Wang, Qun; Huang, Tao; Shu, Xin; Zhao, Shi-Gang; Liang, Yu; Muhammad, Tahir; Gao, Fei; Zhao, Han; Liu, Hong-Bin

2018-01-01

Polycystic ovary syndrome (PCOS) is a heterogeneous disorder characterized by chronic ovulatory dysfunction, hyperandrogenism, and polycystic ovaries. Wilms' tumor 1 (WT1) encoding a transcription factor involved in the differentiation of granulosa cells (GCs) regulates androgen receptor in the development of male genitalia. However, the expression pattern and possible role of WT1 in ovaries of PCOS patients are still unknown. GCs from 95 PCOS patients (PCOS group) and 62 healthy controls (control group) were isolated. The expression of WT1 in GCs was quantified using the reverse transcription-polymerase chain reaction. The correlation between WT1 expression and clinical characteristics was evaluated in PCOS patients. WT1 expression was increased in PCOS patients compared with the normal controls. The expression of WT1 was moderately correlated with testosterone (r = 0.334, p = 0.001) and luteinizing hormone (r = 0.357, p = 0.001) levels and the antral follicle counts (r = 0.337, p = 0.001). Our study provided novel insights into the relationship between hyperandrogenism and polycystic ovaries of PCOS and WT1. © 2018 S. Karger AG, Basel.

Simultaneous extraction and depolymerization of fucoidan from Sargassum muticum in aqueous media.

PubMed

Balboa, Elena M; Rivas, Sandra; Moure, Andrés; Domínguez, Herminia; Parajó, Juan Carlos

2013-11-21

The biomass components of the invasive seaweed Sargassum muticum were fractionated to allow their separate valorization. S. muticum (Sm) and the solid residue remaining after alginate extraction of this seaweed (AESm) were processed with hot, compressed water (hydrothermal processing) to assess the effects of temperature on fucoidan solubilization. Fucose-containing oligosaccharides were identified as reaction products. Operating under optimal conditions (170 °C), up to 62 and 85 wt% of the dry mass of Sm and AESm were solubilized, respectively. The reaction media were subjected to precipitation, nanofiltration and freeze-drying. The dried products contained 50% and 85% of the fucoidan present in Sm and AESm, respectively; together with other components such as phenolics and inorganic components. The saccharidic fraction, accounting for up to 35% of the dried extracts, contained fucose as the main sugar, and also galactose, xylose, glucose and mannose. The concentrates were characterized for antioxidant activity using the TEAC assay.

Adsorption of polyethylene glycol (PEG) onto cellulose nano-crystals to improve its dispersity.

PubMed

Cheng, Dong; Wen, Yangbing; Wang, Lijuan; An, Xingye; Zhu, Xuhai; Ni, Yonghao

2015-06-05

In this work, the adsorption of polyethylene glycol (PEG) onto cellulose nano-crystals (CNC) was investigated for preparing re-dispersible dried CNC. Results showed that the re-dispersity of CNC in water can be significantly enhanced using a PEG1000 dosage of 5wt% (based on the dry weight of CNC). The elemental analysis confirmed the adsorption of PEG onto the CNC surface. Transmission electron microscopy (TEM) was used to characterize the dry powder and indicated that the irreversible agglomeration of CNC after drying was essentially eliminated based on the PEG adsorption concept. Thermo-gravimetric analysis (TGA) and X-ray diffraction (XRD) suggested that CNC crystallinity and thermal stability were not affected by the adsorption of PEG. Thus, the adsorption of PEG has great potential for producing re-dispersible powder CNC. Copyright © 2015 Elsevier Ltd. All rights reserved.

Simultaneous Extraction and Depolymerization of Fucoidan from Sargassum muticum in Aqueous Media

PubMed Central

Balboa, Elena M.; Rivas, Sandra; Moure, Andrés; Domínguez, Herminia; Parajó, Juan Carlos

2013-01-01

The biomass components of the invasive seaweed Sargassum muticum were fractionated to allow their separate valorization. S. muticum (Sm) and the solid residue remaining after alginate extraction of this seaweed (AESm) were processed with hot, compressed water (hydrothermal processing) to assess the effects of temperature on fucoidan solubilization. Fucose-containing oligosaccharides were identified as reaction products. Operating under optimal conditions (170 °C), up to 62 and 85 wt% of the dry mass of Sm and AESm were solubilized, respectively. The reaction media were subjected to precipitation, nanofiltration and freeze-drying. The dried products contained 50% and 85% of the fucoidan present in Sm and AESm, respectively; together with other components such as phenolics and inorganic components. The saccharidic fraction, accounting for up to 35% of the dried extracts, contained fucose as the main sugar, and also galactose, xylose, glucose and mannose. The concentrates were characterized for antioxidant activity using the TEAC assay. PMID:24284426

The Oxidation of AlN in Dry and Wet Oxygen

NASA Technical Reports Server (NTRS)

Opila, Elizabeth; Humphrey, Donald; Jacobson, Nathan; Yoshio, Tetsuo; Oda, Kohei

1998-01-01

The oxidation kinetics of AlN containing 3.5 wt% Y2O3 were studied by thermogravimetric analysis in dry oxygen and 10% H2O/balance oxygen at temperatures between 1000 and 1200 C for times between 48 and 100 h. The oxidation kinetics for AlN in dry oxygen were parabolic and of approximately the same magnitude and temperature dependence as other alumina forming materials. In this case, diffusion of oxygen and/or aluminum through the alumina scale is the rate limiting mechanism. The oxidation kinetics for AlN in wet oxygen were nearly linear and much more rapid than rates observed in dry oxygen. Numerous micropores were observed in the alumina formed on AIN in wet oxygen. These pores provide a fast path for oxygen transport. The linear kinetics observed in this case suggest that the interface reaction rate of AlN with wet oxygen is the oxidation rate limiting step.

A hydrogen transient nuclear magnetic resonance sensor for industrial drying applications

DOE Office of Scientific and Technical Information (OSTI.GOV)

Nicholls, C.

1990-01-01

It has been estimated that industrial non-paper drying processes consume {approximately}0.8 quad (i.e. 8 {times} 10{sup 14} BTU) of energy per year in the United States, representing {approximately}5% of total industrial consumption. If improved technologies could be used to increase the efficiencies of the drying process and hence produce a 2% reduction in energy consumption, the energy savings would be 0.016 quad per year, or {approximately}2.5 million bbl of crude oil. DOE studies indicated that the most attractive R D target to aid in achieving these savings was an advanced moisture sensor, capable of application to a wide variety ofmore » drying processes. To meet these objectives the sensor should accurately monitor product moisture content over the range 2--35 % wt moisture (wb) and be usable at temperatures up to 350{degree}F. 22 refs., 11 figs., 1 tab.« less

HIV-1 replication in cell lines harboring INI1/hSNF5 mutations.

PubMed

Sorin, Masha; Yung, Eric; Wu, Xuhong; Kalpana, Ganjam V

2006-08-31

INI1/hSNF5 is a cellular protein that directly interacts with HIV-1 integrase (IN). It is specifically incorporated into HIV-1 virions. A dominant negative mutant derived from INI1 inhibits HIV-1 replication. Recent studies indicate that INI1 is associated with pre-integration and reverse transcription complexes that are formed upon viral entry into the target cells. INI1 also is a tumor suppressor, biallelically deleted/mutated in malignant rhabdoid tumors. We have utilized cell lines derived from the rhabdoid tumors, MON and STA-WT1, that harbor either null or truncating mutations of INI1 respectively, to assess the effect of INI1 on HIV-1 replication. We found that while HIV-1 virions produced in 293T cells efficiently transduced MON and STA-WT1 cells, HIV-1 particle production was severely reduced in both of these cells. Reintroduction of INI1 into MON and STA-WT1 significantly enhanced the particle production in both cell lines. HIV-1 particles produced in MON cells were reduced for infectivity, while those produced in STA-WT1 were not. Further analysis indicated the presence of INI1 in those virions produced from STA-WT1 but not from those produced from MON cells. HIV-1 produced in MON cells were defective for synthesis of early and late reverse transcription products in the target cells. Furthermore, virions produced in MON cells were defective for exogenous reverse transcriptase activity carried out using exogenous template, primer and substrate. Our results suggest that INI1-deficient cells exhibit reduced particle production that can be partly enhanced by re-introduction of INI1. Infectivity of HIV-1 produced in some but not all INI1 defective cells, is affected and this defect may correlate to the lack of INI1 and/or some other proteins in these virions. The block in early events of virion produced from MON cells appears to be at the stage of reverse transcription. These studies suggest that presence of INI1 or some other host factor in virions and reverse transcription complexes may be important for early events of HIV-1 replication.

Calpastatin overexpression impairs postinfarct scar healing in mice by compromising reparative immune cell recruitment and activation.

PubMed

Wan, Feng; Letavernier, Emmanuel; Le Saux, Claude Jourdan; Houssaini, Amal; Abid, Shariq; Czibik, Gabor; Sawaki, Daigo; Marcos, Elisabeth; Dubois-Rande, Jean-Luc; Baud, Laurent; Adnot, Serge; Derumeaux, Geneviève; Gellen, Barnabas

2015-12-01

The activation of the calpain system is involved in the repair process following myocardial infarction (MI). However, the impact of the inhibition of calpain by calpastatin, its natural inhibitor, on scar healing and left ventricular (LV) remodeling is elusive. Male mice ubiquitously overexpressing calpastatin (TG) and wild-type (WT) controls were subjected to an anterior coronary artery ligation. Mortality at 6 wk was higher in TG mice (24% in WT vs. 44% in TG, P < 0.05) driven by a significantly higher incidence of cardiac rupture during the first week post-MI, despite comparable infarct size and LV dysfunction and dilatation. Calpain activation post-MI was blunted in TG myocardium. In TG mice, inflammatory cell infiltration and activation were reduced in the infarct zone (IZ), particularly affecting M2 macrophages and CD4(+) T cells, which are crucial for scar healing. To elucidate the role of calpastatin overexpression in macrophages, we stimulated peritoneal macrophages obtained from TG and WT mice in vitro with IL-4, yielding an abrogated M2 polarization in TG but not in WT cells. Lymphopenic Rag1(-/-) mice receiving TG splenocytes before MI demonstrated decreased T-cell recruitment and M2 macrophage activation in the IZ day 5 after MI compared with those receiving WT splenocytes. Calpastatin overexpression prevented the activation of the calpain system after MI. It also impaired scar healing, promoted LV rupture, and increased mortality. Defective scar formation was associated with blunted CD4(+) T-cell and M2-macrophage recruitment. Copyright © 2015 the American Physiological Society.

Cell type of origin as well as genetic alterations contribute to breast cancer phenotypes

PubMed Central

West, William W.; Qiu, Fang; Band, Hamid; Band, Vimla

2015-01-01

Breast cancer is classified into different subtypes that are associated with different patient survival outcomes, underscoring the importance of understanding the role of precursor cell and genetic alterations in determining tumor subtypes. In this study, we evaluated the oncogenic phenotype of two distinct mammary stem/progenitor cell types designated as K5+/K19− or K5+/K19+ upon introduction of identical combinations of oncogenes-mutant H-Ras (mRas) and mutant p53 (mp53), together with either wild-type ErbB2(wtErbB2) or wild-type EGFR (wtEGFR). We examined their tumor forming and metastasis potential, using both in-vitro and in-vivo assays. Both the combinations efficiently transformed K5+/K19− or K5+/K19+ cells. Xenograft tumors formed by these cells were histologically heterogeneous, with variable proportions of luminal, basal-like and claudin-low type components depending on the cell types and oncogene combinations. Notably, K5+/K19− cells transformed with mRas/mp53/wtEGFR combination had a significantly longer latency for primary tumor development than other cell lines but more lung metastasis incidence than same cells expressing mRas/mp53/wtErbB2. K5+/K19+ cells exhibit shorter overall tumor latency, and high metastatic potential than K5+/K19− cells, suggesting that these K19+ progenitors are more susceptible to oncogenesis and metastasis. Our results suggest that both genetic alterations and cell type of origin contribute to oncogenic phenotype of breast tumors. PMID:25940703

The influence of bioglass nanoparticles on the biodegradation and biocompatibility of poly (3-hydroxybutyrate) scaffolds.

PubMed

Hajiali, Hadi; Hosseinalipour, Mohammad; Karbasi, Saeed; Shokrgozar, Mohammad Ali

2012-11-01

Nanocomposite scaffolds have been developed in order to achieve better mechanical and physiological properties in bone tissue engineering applications. In this study, reinforced poly (3-hydroxybutyrate) (PHB) composite scaffolds made with different weight ratios of nanobioglass (0, 2.5, 5, 7.5, and 10 wt%) and various porosities (70, 80 and 90 wt% of NaCl) were prepared by the salt leaching process. The scaffolds were placed in a PBS solution and their weight loss was measured. The biocompatibility of samples was examined in vitro using the MG63 cell line by indirect test, cell proliferation, and alkaline phosphatase (ALP) assays. Cell attachment on the surface of the scaffolds was observed by scanning electron microscopy (SEM). The biodegradation results showed that increasing the volume fraction of porosity and concentration of bioglass nanoparticles enhanced the weight loss of the scaffolds. The cell study demonstrated that a certain concentration of nanobioglass (7.5 wt%) in the scaffolds can significantly improve cell proliferation, inducing better osteoconductivity, compared to that of the pure PHB scaffolds and controls. In addition, the SEM results showed high cell attachment on these samples. All these factors indicate that samples with 7.5 wt% nanobioglass are a promising scaffold for bone tissue engineering.

Morphological and mechanical characterization of composite bone cement containing polymethylmethacrylate matrix functionalized with trimethoxysilyl and bioactive glass.

PubMed

Puska, Mervi; Moritz, Niko; Aho, Allan J; Vallittu, Pekka K

2016-06-01

Medical polymers of biostable nature (e.g. polymethylmetacrylate, PMMA) are widely used in various clinical applications. In this study, novel PMMA-based composite bone cement was prepared. Bioactive glass (BAG) particulate filler (30wt%) was added to enhance potentially the integration of bone to the cement. The polymer matrix was functionalized with trimethoxysilyl to achieve an interfacial bond between the matrix and the fillers of BAG. The amount of trimethoxysilyl in the monomer system varied from 0 to 75wt%. The effects of dry and wet (simulated body fluid, SBF at +37°C for 5 weeks) conditions were investigated. In total, 20 groups of specimens were prepared. The specimens were subjected to a destructive mechanical test in compression. Scanning electron microscopy (SEM) and micro-computed tomography (micro-CT) were used to study the surface and the three-dimensional morphology of the specimens. The results of the study indicated that the addition of trimethoxysilyl groups led to the formation of a hybrid polymer matrix which, in lower amounts (<10wt% of total weight), did not significantly affect the compression properties. However, when the specimens stored in dry and wet conditions were compared, the water sorption increased the compression strength (~5-10MPa per test group). At the same time, the water sorption also caused an evident porous structure formation for the specimens containing BAG and siloxane formation in the hybrid polymer matrix. Copyright © 2015 Elsevier Ltd. All rights reserved.

Modification of bone graft by blending with lecithin to improve hydrophilicity and biocompatibility.

PubMed

Wang, Y; Cui, F Z; Jiao, Y P; Hu, K; Fan, D D

2008-03-01

Lecithin was blended to improve the hydrophilicity and biocompatibility of bone graft containing poly(l-lactic acid) (PLLA). Solution blending and freeze drying were used to fabricate symmetrical scaffolds containing different percentages of lecithin (lecithin: PLLA = 0, 5, 10 wt%). Scanning electron microscopy showed that the scaffolds maintained the three-dimensional porous structure. A water uptake experiment proved the significant improvement of hydrophilicity of the blend scaffold. With the addition of lecithin, the compressive strength and compressive modulus decreased. When the weight ratio of lecithin to PLLA was up to 10%, the compressive strength was still more than the lower limit of natural cancellous bone. To test the biocompatibility of the scaffolds, cell culture in vitro and subcutaneous implantation in vivo were performed. MC3T3-E1 preosteoblastic cells were cultured on the scaffolds for 7 days. Methylthiazol tetrazolium assay and laser scanning confocal microscopy were used to exhibit proliferation and morphology of the cells. The subcutaneous implantation in rats tested inflammatory response to the scaffolds. The results proved the better biocompatibility and milder inflammatory reactions of the blend scaffold (lecithin: PLLA = 5%) compared with the scaffold without lecithin. The modified scaffold containing lecithin is promising for bone tissue engineering.

Ethanol production in a membrane bioreactor: pilot-scale trials in a corn wet mill.

PubMed

Escobar, J M; Rane, K D; Cheryan, M

2001-01-01

Pilot plant trials were conducted in a corn wet mill with a 7000-L membrane recycle bioreactor (MRB) that integrated ceramic microfiltration membranes in a semi-closed loop configuration with a stirred-tank reactor. Residence times of 7.5-10 h with ethanol outputs of 10-11.5% (v/v) were obtained when the cell concentration was 60-100 g/L dry wt of yeast, equivalent to about 10(9)-10(10) cells/mL. The performance of the membrane was dependent on the startup mode and pressure management techniques. A steady flux of 70 L/(m2 x h) could be maintained for several days before cleaning was necessary. The benefits of the MRB include better productivity; a clear product stream containing no particulates or yeast cells, which should improve subsequent stripping and distillation operations; and substantially reduced stillage handling. The capital cost of the MRB is $21-$34/(m3 x yr) ($0.08-$0.13/[gal x yr]) of ethanol capacity. Operating cost, including depreciation, energy, membrane replacement, maintenance, labor, and cleaning, is $4.5-9/m3 ($0.017-$0.034/gal) of ethanol.

The hematopoietic tumor suppressor interferon regulatory factor 8 (IRF8) is upregulated by the antimetabolite cytarabine in leukemic cells involving the zinc finger protein ZNF224, acting as a cofactor of the Wilms' tumor gene 1 (WT1) protein.

PubMed

Montano, Giorgia; Ullmark, Tove; Jernmark-Nilsson, Helena; Sodaro, Gaetano; Drott, Kristina; Costanzo, Paola; Vidovic, Karina; Gullberg, Urban

2016-01-01

The transcription factor interferon regulatory factor-8 (IRF8) is highly expressed in myeloid progenitors, while most myeloid leukemias show low or absent expression. Loss of IRF8 in mice leads to a myeloproliferative disorder, indicating a tumor-suppressive role of IRF8. The Wilms tumor gene 1 (WT1) protein represses the IRF8-promoter. The zinc finger protein ZNF224 can act as a transcriptional co-factor of WT1 and potentiate the cytotoxic response to the cytostatic drug cytarabine. We hypothesized that cytarabine upregulates IRF8 and that transcriptional control of IRF8 involves WT1 and ZNF224. Treatment of leukemic K562 cells with cytarabine upregulated IRF8 protein and mRNA, which was correlated to increased expression of ZNF224. Knock down of ZNF224 with shRNA suppressed both basal and cytarabine-induced IRF8 expression. While ZNF224 alone did not affect IRF8 promoter activity, ZNF224 partially reversed the suppressive effect of WT1 on the IRF8 promoter, as judged by luciferase reporter experiments. Coprecipitation revealed nuclear binding of WT1 and ZNF224, and by chromatin immunoprecipitation (ChIP) experiments it was demonstrated that WT1 recruits ZNF224 to the IRF8 promoter. We conclude that cytarabine-induced upregulation of the IRF8 in leukemic cells involves increased levels of ZNF224, which can counteract the repressive activity of WT1 on the IRF8-promoter. Copyright © 2015 Elsevier Ltd. All rights reserved.

Adhesion of a fluorinated poly(amic acid) with stainless steel surfaces

NASA Astrophysics Data System (ADS)

Jung, Youngsuk; Song, Sunjin; Kim, Sangmo; Yang, Yooseong; Chae, Jungha; Park, Tai-Gyoo; Dong Cho, Myung

2013-01-01

The authors elucidate an origin and probable mechanism of adhesion strength change at an interface of fluorinated poly(amic acid) and stainless steel. Fluorination provides favorable delamination with release strength weaker than 0.08 N/mm from a metal surface, once the amount of residual solvent becomes less than 35 wt. %. However, the release strength critically depends on film drying temperature. Characterization on stainless steel surfaces and thermodynamic analyses on wet films reveal a drying temperature of 80 °C fosters interaction between the metal oxides at stainless steel surface and the free electron donating groups in poly(amic acid).

Betaine synthesis and accumulation in barley during field water-stress

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hitz, W.D.; Ladyman, J.A.R.; Hanson, A.D.

1982-01-01

The timing and extent of betaine accumulation by mature leaves of barley (Hordeum vulgare L.) were followed in irrigated (I) and non-irrigated (N-I) plots under rain-shelters. In the N-I crop, leaf water potential (/sup psi/leaf) began to fall at the five-leaf stage, continued to drop steadily until maturity, and reached a minimum of about -35 bars. Betaine accumulation started in the N-I crop about a week after the decline in /sup psi/leaf began and continued until about 10 days post-anthesis. The maximum betaine concentration attained by N-I leaves (100 ..mu..mol/g dry wt) was three times that in I leaves. Betainemore » accumulation by upper leaves was due mainly to de novo synthesis in these leaves, because: (1) there was little /sup 14/C-import into upper leaves when (/sup 14/C)betaine was applied to lower leaves, and (2) attached upper leaves of N-I plants rapidly converted supplied (/sup 14/C)ethanolamine to (/sup 14/C)betaine during the peak period of betaine accumulation. Phosphatidylcholine (PC) behaved as an intermediate in the conversion of (/sup 14/C)ethanolamine to betaine. The estimated peak metabolic cost of betaine biosynthesis via PC by stressed leaves (about 2 mg hexose/g dry wt/day) approached the cost of protein turnover in the same leaves (3 to 5 mg hexose/g dry wt/day) as estimated from (/sup 3/H) lysine incorporation. In N-I plants, cessation of betaine synthesis preceded the onset of senescence by several days, indicating that continuous betaine production is not mandatory for leaf function at lowered /sup psi/leaf. These field results are consistent with an adaptive value for betaine accumulation in barley during prolonged water stress. A search for genetic variation in betaine-accumulating potential in barley is now warranted.« less

Trace element concentrations in livers of polar bears from two populations in Northern and Western Alaska.

PubMed

Kannan, Kurunthachalam; Agusa, Tetsuro; Evans, Thomas J; Tanabe, Shinsuke

2007-10-01

Concentrations of 20 trace elements (V, Cr, Mn, Co, Cu, Zn, Rb, Sr, Mo, Ag, Cd, In, Sn, Sb, Cs, Ba, Hg, Tl, Pb, and Bi) were measured in livers of polar bears (Ursus maritimus) collected from Northern and Western Alaska from 1993 to 2002 to examine differences in the profiles of trace metals between the Beaufort Sea (Northern Alaska) and the Chukchi Sea (Western Alaska) subpopulations in Alaska. Among the trace elements analyzed, concentrations of Cu (50-290 microg/g, dry wt) in polar bear livers were in the higher range of values that have been reported for marine mammals. Concentrations of Hg in polar bears varied widely, from 3.5 to 99 microg/g dry wt, and the mean concentrations in polar bears were comparable to concentrations reported previously for several other species of marine mammals. Mean concentrations of Pb and Cd were 0.67 and 1.0 microg/g dry wt, respectively; these concentrations were lower than levels reported elsewhere for polar bears from Greenland and Canada. Age- and gender-related variations in the concentrations of trace elements in our polar bears were minimal. Concentrations of Hg decreased slowly in samples collected during 1993-2002, whereas Cd and Pb concentrations were found to be stable or slowly increasing, in the livers of Alaskan polar bears. Concentrations of Ag, Bi, Ba, Cu, and Sn were significantly higher in the Chukchi Sea subpopulation than in the Beaufort Sea subpopulation. Concentrations of Hg were significantly higher in the Beaufort Sea subpopulation than in the Chukchi Sea subpopulation. Differences in the profiles and concentrations of Hg, Ag, Bi, Ba, Cu, and Sn suggest that the sources of exposure to these trace elements between Western and Northern Alaskan polar bears are different, in agreement with findings reported earlier for several organic contaminants.

Evaluation of the metabolic fate of munitions material (TNT & RDX) in plant systems and initial assessment of material interaction with plant genetic material. Validation of the metabolic fate of munitions materials (TNT, RDX) in mature crops

DOE Office of Scientific and Technical Information (OSTI.GOV)

Fellows, R.J.; Harvey, S.D.; Cataldo, D.A.

1995-09-01

The goals of this effort were to confirm and expand data related to the behavior and impacts of munitions residues upon human food chain components. Plant species employed included corn (Zea mays), alfalfa (Medicago sativa). spinach (Spinacea oleraceae), and carrot (Daucus carota). Plants were grown from seed to maturity (70 to 120 days) in a low-fertility soil (Burbank) amended with either {sup 14}C-TNT or {sup 14}C-RDX at which time they were harvested and analyzed for munitions uptake, partitioning, and chemical form of the munition or munition-metabolite. All four of the plant species used in this study accumulated the {sup 14}C-TNT-more » and RDX-derived label. The carrot, alfalfa, and corn demonstrated a higher percentage of label retained in the roots (62, 73, and 83% respectively). The spinach contained less activity in its root (36%) but also contained the highest TNT specific activity observed (>4600 jig TNT equivalents/g dry wt.). The specific uptake values of RDX for the spinach and alfalfa were comparable to those previously reported for wheat and bean (314 to 590 {mu}g RDX-equivalents/g dry wt. respectively). An exception to this may be the carrot where the specific activity was found to exceed 4200 {mu}g RDX-equivalents/g dry wt. in the shoot. The total accumulation of TNT by the plants ranged from 1.24% for the spinach to 2.34% for the carrot. The RDX plants ranging from 15% for the spinach to 37% for the carrot. There was no identifiable TNT or amino dinitrotoluene (ADNT) isomers present in the plants however, the parent RDX compound was found at significant levels in the shoot of alfalfa (> 1 80 {mu}g/g) and corn (>18 {mu}g/g).« less

Effect of Organic Material on Mechanical, Hydrological, and Microstructural Properties of Mudstones

NASA Astrophysics Data System (ADS)

Altobelli, M. A.; Reece, J. S.

2016-12-01

In this research we analyze the influence of organic material on the mechanical and flow properties of mudstones. We uniformly mix peat, milled and harvested by Bord na Móna from the surface of bogs in Ireland, with natural mudstone from Site C0011 in the Nankai Trough, offshore Japan, obtained during Integrated Ocean Drilling Program Expedition 322. The mudstone had previously been disaggregated into a homogeneous dry powder of clay- and silt-sized particles. The peat is ground and dry-sieved to achieve a similar particle size distribution as the mudstone (< 63 micrometers). In order to understand the mechanical and hydrological processes affected by peat, we prepare dry peat-mudstone mixtures with three different peat concentrations: 0 wt%, 5 wt%, and 10 wt%. Then, these peat - mudstone mixtures are saturated with deionized water at a water content of 109%, formed into stable slurries, and uniaxially compressed to an axial stress of 100 kPa using resedimentation, a method that simulates the natural behavior of deposition and burial in the laboratory under controlled conditions. How the organic material interacts with the mudstone matrix and pore fluid under compression influences the physical properties of the mudstones such as porosity, compressibility, and permeability; all of which are measured in the resedimentation experiments. We will also analyze the microstructural changes as a function of peat concentration using a petrographic microscope and scanning electron microscope. Due to the fibrous and absorbent nature of peat, we anticipate the peat to force tightly packed clay particles in the mudstone apart resulting in a looser microstructure and increased porosity, and thus, a higher compressibility and permeability. Understanding the controls on the mechanical and flow properties of hydrocarbon-bearing, fine-grained formations is crucial for exploration and successful production from hydrocarbon reservoirs. Additionally, this study has large implications for soil water storage and soil amendment to improve plant growth and health in clayey soils.

Macrophage-derived insulin-like growth factor-1 affects influenza vaccine efficacy through the regulation of immune cell homeostasis.

PubMed

Yoon, Il-Sub; Park, Hyelim; Kwak, Hye-Won; Woo Jung, Yong; Nam, Jae-Hwan

2017-08-24

The level of antibody production induced by a vaccine involves a variety of host factors. One of these, insulin-like growth factor-1 (IGF-1), plays an important role in lymphocyte maturation and antibody expression. Here, we investigated the role of macrophage-derived IGF-1 in the induction of influenza vaccine-specific antibodies using macrophage-derived IGF-1 gene knockout (MIKO) mice. The titers of vaccine-specific total immunoglobulin G (IgG) and IgG1 after immunization were about two- to fourfold lower in MIKO mice than in WT mice. Moreover, MIKO mice showed a relatively weak booster effect of repeated immunization. In contrast, antigen-nonspecific total IgG was about threefold higher in MIKO mice than in WT mice. After viral challenge, the viral titer and the pathological damage in lungs of MIKO mice were higher than those in WT mice despite vaccination. Interestingly, the proportions of proinflammatory immune cells including M1 macrophages, Th1 and Th17 cells was higher in unvaccinated MIKO mice than in unvaccinated WT mice. This suggests that nonspecific activation of immune cells may paradoxically impair the response to the vaccine. In addition, although the proportions of T follicular helper (Tfh) cells and GL-7 + germinal center (GC) B cells were higher in MIKO mice than in WT mice, the population of CD138 + B220 + antibody-secreting plasmablasts was lower in MIKO mice, which may be a cause of the low influenza-specific antibody titer in MIKO mice. Taken together, these results suggest that macrophage-derived IGF-1 might play an important role in the vaccine-triggered immune response by regulating immune cell homeostasis. Copyright © 2017 Elsevier Ltd. All rights reserved.

Adenosine A1 receptors contribute to immune regulation after neonatal hypoxic ischemic brain injury.

PubMed

Winerdal, Max; Winerdal, Malin E; Wang, Ying-Qing; Fredholm, Bertil B; Winqvist, Ola; Ådén, Ulrika

2016-03-01

Neonatal brain hypoxic ischemia (HI) often results in long-term motor and cognitive impairments. Post-ischemic inflammation greatly effects outcome and adenosine receptor signaling modulates both HI and immune cell function. Here, we investigated the influence of adenosine A1 receptor deficiency (A1R(-/-)) on key immune cell populations in a neonatal brain HI model. Ten-day-old mice were subjected to HI. Functional outcome was assessed by open locomotion and beam walking test and infarction size evaluated. Flow cytometry was performed on brain-infiltrating cells, and semi-automated analysis of flow cytometric data was applied. A1R(-/-) mice displayed larger infarctions (+33%, p < 0.05) and performed worse in beam walking tests (44% more mistakes, p < 0.05) than wild-type (WT) mice. Myeloid cell activation after injury was enhanced in A1R(-/-) versus WT brains. Activated B lymphocytes expressing IL-10 infiltrated the brain after HI in WT, but were less activated and did not increase in relative frequency in A1R(-/-). Also, A1R(-/-) B lymphocytes expressed less IL-10 than their WT counterparts, the A1R antagonist DPCPX decreased IL-10 expression whereas the A1R agonist CPA increased it. CD4(+) T lymphocytes including FoxP3(+) T regulatory cells, were unaffected by genotype, whereas CD8(+) T lymphocyte responses were smaller in A1R(-/-) mice. Using PCA to characterize the immune profile, we could discriminate the A1R(-/-) and WT genotypes as well as sham operated from HI-subjected animals. We conclude that A1R signaling modulates IL-10 expression by immune cells, influences the activation of these cells in vivo, and affects outcome after HI.

Parp1 activation in mouse embryonic fibroblasts promotes Pol β-dependent cellular hypersensitivity to alkylation damage

PubMed Central

Jelezcova, Elena; Trivedi, Ram N.; Wang, Xiao-hong; Tang, Jiang-bo; Brown, Ashley R.; Goellner, Eva M.; Schamus, Sandy; Fornsaglio, Jamie L.; Sobol, Robert W.

2010-01-01

Alkylating agents induce cell death in wild-type (WT) mouse embryonic fibroblasts (MEFs) by multiple mechanisms, including apoptosis, autophagy and necrosis. DNA polymerase β (Pol β) knockout (KO) MEFs are hypersensitive to the cytotoxic effect of alkylating agents, as compared to WT MEFs. To test the hypothesis that Parp1 is preferentially activated by methyl methanesulfonate (MMS) exposure of Pol β KO MEFs, we have examined the relationship between Pol β expression, Parp1 activation and cell survival following MMS exposure in a series of WT and Pol β deficient MEF cell lines. Consistent with our hypothesis, we observed elevated Parp1 activation in Pol β KO MEFs as compared to matched WT MEFs. Both the MMS-induced activation of Parp1 and the MMS-induced cytoxicity of Pol β KO MEFs are attenuated by pre-treatment with the Parp1/Parp2 inhibitor PJ34. Further, elevated Parp1 activation is observed following knockdown (KD) of endogenous Pol β, as compared to WT cells. Pol β KD MEFs are hypersensitive to MMS and both the MMS-induced hypersensitivity and Parp1 activation is prevented by pre-treatment with PJ34. In addition, the MMS-induced cellular sensitivity of Pol β KO MEFs is reversed when Parp1 is also deleted (Pol β/Parp1 double KO MEFs) and we observe no MMS sensitivity differential between Pol β/Parp1 double KO MEFs and those that express recombinant mouse Pol β. These studies suggest that Parp1 may function as a sensor of BER to initiate cell death when BER is aborted or fails. Parp1 may therefore function in BER as a tumor suppressor by initiating cell death and preventing the accumulation of cells with chromosomal damage due to a BER defect. PMID:20096707

E-cadherin genetic variants predict survival outcome in breast cancer patients.

PubMed

Memni, Hager; Macherki, Yosra; Klayech, Zahra; Ben-Haj-Ayed, Ahlem; Farhat, Karim; Remadi, Yassmine; Gabbouj, Sallouha; Mahfoudh, Wijden; Bouzid, Nadia; Bouaouina, Noureddine; Chouchane, Lotfi; Zakhama, Abdelfattah; Hassen, Elham

2016-11-16

E-cadherin is a major component of adherens junctions that regulates cell shape and maintains tissue integrity. A complete loss or any decrease in cell surface expression of E-cadherin will interfere with the cell-to-cell junctions' strength and leads to cell detachment and escape from the primary tumor site. In this prospective study, three functional single nucleotide polymorphisms (-347G/GA, rs5030625; -160C/A, rs16260; +54C/T, rs1801026), were found to modulate E-cadherin expression. 577 DNA samples from breast cancer (BC) cases were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). We detected no significant correlations between each polymorphism and the clinical parameters of the patients whereas the GACC haplotype was significantly associated with low SBR grading. Overall survival analysis showed that both -347G/G and +54C/C wild (wt) genotypes had a significantly worse effect compared to the other genotypes (non-wt). Moreover, carrying simultaneously both the -347 and +54 wt genotypes confers a significantly higher risk of death. However, with metastatic recurrence, the death-rate was null in patients carrying the non-wt genotypes, and attained 37% in those carrying the wt genotype. A multivariate analysis showed that these two polymorphisms are independent prognostic factors for overall survival in BC patients. Our results support the fact that E-cadherin genetic variants control disease severity and progression and could be a marker of disease outcome. These findings could be useful in selecting patients that should be monitored differently.

Stability of carbonated basaltic melt at the base of the Earth's upper mantle

NASA Astrophysics Data System (ADS)

Ghosh, S.; Litasov, K.; Ohtani, E.; Suzuki, A.

2006-12-01

Seismological observations of low velocity zones (LVZ) at the top of the 410-km discontinuity reveal possible existence of dense melt at this boundary (e.g. Reveanugh and Sipkin, 1994). Density measurements of anhydrous basaltic melts indicate that it is denser than surrounding mantle near 410-km depth (Ohtani and Maeda, 2001). However, melting temperature of peridotite is much higher than about 1400°C, estimated at 410-km depth. It has been shown recently that hydrous basaltic melt containing up to 2 wt.% H2O is denser than peridotite atop 410-km and therefore can be accumulated at the base of the upper mantle (Sakamaki et al., 2006). CO2 is another major volatile component in the mantle and it could be also important for explanation of LVZ near 410 km. In the present study, we have measured the density of carbonated basaltic melt at high pressures and high temperatures and discussed its possible stability at the base of the upper mantle. The density of the melt was determined using sink/float technique. The starting material was synthetic MORB glass. 5 and 10 wt.% CO2 was added to the glass as CaCO3 and Na2CO3, adjusting to proportions of related oxides. Experiments were carried out at 16-22 GPa and 2200-2300°C using a multianvil apparatus at Tohoku University, Japan. We observed neutral buoyancy of diamond density marker in MORB + 5 wt.% CO2 at 18 GPa and 2300°C, whereas, diamond was completely dissolved in the carbonated MORB melt containing 10 wt.% CO2 in 0.5-1 minute experiments. Based on the buoyancy test, the density of the carbonated basaltic melt, containing 5 wt.% CO2, is 3.56 g/cm3 at 18 GPa and 2300°C using an equation of state of diamond. To calculate the bulk modulus we assume that the pressure derivative of the isothermal bulk modulus is the same as that of the dry MORB melt, dKT/dP=5.0 and zero-pressure partial molar volume of CO2 is 32 cm3/mol (based on low-pressure experiments on carbonated basaltic melts and carbonatites, e.g. Dobson et al., 1996; Liu and Lange, 2003). Accordingly, the isothermal bulk modulus (KT) of the carbonated MORB melt containing 5 wt.% CO2 calculated using the Birch-Murnaghan equation of state is 16.3 ± 1 GPa. This value is close to that of dry MORB (KT=18 GPa) and indicates that addition of 5 wt.% CO2 to basaltic melt has minor influence on its compressibility. Density of MORB + 5 wt.% CO2 is almost same with the density of MORB + 2 wt.% H2O at 15-20 GPa. Comparison of the density of carbonated basaltic melt with PREM density profile at 1600°C indicates that it is buoyant above the 410 km discontinuity in the mantle only if it contains more than about 5 wt.% CO2.

Enhanced Mulberroside A Production from Cell Suspension and Root Cultures of Morus alba Using Elicitation.

PubMed

Komaikul, Jukrapun; Kitisripanya, Tharita; Tanaka, Hiroyuki; Sritularak, Boonchoo; Putalun, Waraporn

2015-07-01

Morus alba L. has been used in Asian traditional medicine as an anti-inflammatory, anti-asthmatic, anthelmintic and as a whitening agent in cosmetic products. Mulberroside A is the major active compound from M. alba root bark. In this study, cell suspension and root cultures of M. alba were established, and the effect of the elicitors on the enhancement of mulberroside A production in M. alba was investigated. The cell suspension and root cultures of M. alba were exposed to elicitors and then mulberroside A contents were determined by an indirect competitive ELISA method. High levels of mulberroside A were obtained by addition of 100 and 200 μM salicylic acid with 24 h exposure time in cell suspension cultures (37.9 ± 1.5 and 34.0 ± 4.7 mg/g dry wt., respectively). Furthermore, addition of yeast extract at 2 mg/mL with 24 h exposure time can significantly increase mulberroside A contents from both cell suspension (3.2-fold) and root cultures (6.6-fold). Mulberroside A contents from both cell suspension and root cultures after treatment with elicitors are similar or higher than those found in the intact root and root bark of several years old M. alba. These results indicate that mulberry tissue cultures using the elicitation method are interesting alternative sources for mulberroside A production.

Impaired compensatory beta-cell function and growth in response to high-fat diet in LDL receptor knockout mice

PubMed Central

Oliveira, Ricardo B d; Carvalho, Carolina P d F; Polo, Carla C; Dorighello, Gabriel d G; Boschero, Antônio C; Oliveira, Helena C F d; Collares-Buzato, Carla B

2014-01-01

In this study, we investigated the effect of low density lipoprotein receptor (LDLr) deficiency on gap junctional connexin 36 (Cx36) islet content and on the functional and growth response of pancreatic beta-cells in C57BL/6 mice fed a high-fat (HF) diet. After 60 days on regular or HF diet, the metabolic state and morphometric islet parameters of wild-type (WT) and LDLr−/− mice were assessed. HF diet-fed WT animals became obese and hypercholesterolaemic as well as hyperglycaemic, hyperinsulinaemic, glucose intolerant and insulin resistant, characterizing them as prediabetic. Also they showed a significant decrease in beta-cell secretory response to glucose. Overall, LDLr−/− mice displayed greater susceptibility to HF diet as judged by their marked cholesterolaemia, intolerance to glucose and pronounced decrease in glucose-stimulated insulin secretion. HF diet induced similarly in WT and LDLr−/− mice, a significant decrease in Cx36 beta-cell content as revealed by immunoblotting. Prediabetic WT mice displayed marked increase in beta-cell mass mainly due to beta-cell hypertrophy/replication. Nevertheless, HF diet-fed LDLr−/− mice showed no significant changes in beta-cell mass, but lower islet–duct association (neogenesis) and higher beta-cell apoptosis index were seen as compared to controls. The higher metabolic susceptibility to HF diet of LDLr−/− mice may be explained by a deficiency in insulin secretory response to glucose associated with lack of compensatory beta-cell expansion. PMID:24853046

An analysis of two open reading frames (ORF3 and ORF4) of rat hepatitis E virus genome using its infectious cDNA clones with mutations in ORF3 or ORF4.

PubMed

Tanggis; Kobayashi, Tominari; Takahashi, Masaharu; Jirintai, Suljid; Nishizawa, Tsutomu; Nagashima, Shigeo; Nishiyama, Takashi; Kunita, Satoshi; Hayama, Emiko; Tanaka, Takeshi; Mulyanto; Okamoto, Hiroaki

2018-04-02

Rat hepatitis E virus (ratHEV) genome has four open reading frames (ORFs: ORF1, ORF2, ORF3 and ORF4). The functions of ORF3 and ORF4 are unknown. An infectious cDNA clone (pUC-ratELOMB-131L_wt, wt) and its derivatives including ORF3-defective (ΔORF3) and ORF4-defective (ΔORF4) mutants, were constructed and their full-length RNA transcripts transfected into PLC/PRF/5 cells. ΔORF3 replicated as efficiently as wt in cells. However, ≤1/1000 of the number of progenies were detectable in the culture supernatant of ΔORF3-infected cells compared with wt-infected cells. ORF4 protein was not detectable in ratHEV-infected cells or in the liver tissues of ratHEV-infected rats. No marked differences were noted between wt and ΔORF4 regarding the viral replication and protein expression. ORF3 mutants with proline-to-leucine mutations at amino acids (aa) 93, 96 and/or 98 in ORF3 were constructed and transfected into PLC/PRF/5 cells. Wt and an ORF3 mutant with leucine at aa 98 (ORF3-L98) replicated efficiently (density 1.15-1.16 g/cm 3 ), while ORF3-L93 + L96 exhibited a decreased viral release and banded at 1.26-1.27 g/cm 3 , similar to ΔORF3. In conclusion, the ORF3 protein, especially its proline residues at aa 93 and 96, is essential for the release of membrane-associated ratHEV particles, and ORF4 is unnecessary for the replication of ratHEV. Copyright © 2018 Elsevier B.V. All rights reserved.

Effects of endoplasmic reticulum stressors on maturation and signaling of hemizygous and heterozygous wild-type and mutant forms of KIT.

PubMed

Brahimi-Adouane, Sabrina; Bachet, Jean-Baptiste; Tabone-Eglinger, Séverine; Subra, Frédéric; Capron, Claude; Blay, Jean-Yves; Emile, Jean-François

2013-06-01

Gain of function mutations of KIT are frequent in some human tumors, and are sensible to tyrosine kinase inhibitors. In most tumors, oncogenic mutations are heterozygous, however most in vitro data of KIT activation have been obtained with hemizygous mutation. This study aimed to investigate the maturation and activation of wild-type (WT) and mutant (M) forms of KIT in hemizygous and heterozygous conditions. WT and two types of exon 11 deletions M forms of human KIT were expressed in NIH3T3 cell lines. Membrane expression of KIT was quantified by flow cytometry. Quantification of glycosylated forms of KIT and phosphorylated forms of AKT and ERK were performed by western blot. Simultaneous activation of WT KIT and treatment with endoplasmic reticulum (ER) inhibitors, tunicamycin or brefeldin A induced a complete inhibition of membrane expression of the 145 kDa form of KIT. By contrast activation or ER inhibitors alone, only partly inhibited this form. ER inhibitors also inhibited KIT activation-dependent phosphorylation of AKT and ERK1/2. Brefeldin A induced a complete down regulation of the 145 kDa form in hemizygous M, and induced an intra-cellular accumulation of the 125 kDa form in WT but not in hemizygous M. Heterozygous cells had glycosylation and response to ER inhibitors patterns more similar to WT than to hemizygous M. Phosphorylated AKT was reduced in hemizygous cells in comparison to WT KIT cells and heterozygous cells, and in the presence of brefeldin A in all cell lines. Effects of ER inhibitors are significantly different in hemizygous and heterozygous mutants. Differences in intra-cellular trafficking of KIT forms result in differences in downstream signaling pathways, and activation of PI3K/AKT pathway appears to be tied to the presence of the mature 145 kDa form of KIT at the membrane surface. Copyright © 2012 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

Modulation of Stem Cells Differentiation and Myostatin as an Approach to Counteract Fibrosis in Muscle Dystrophy and Regeneration after Injury

DTIC Science & Technology

2008-03-01

well as the other parallel work with bone marrow cells [ 22 ], the effects were comparatively short- lived, since in our case, the considerable...comparison of myogenic, fibrogenic and adipogenic potential of stem cells from intact and regenerating muscle from mdx, wt and Mst(-/-) mice; b) effects on...sections (not shown). Figure 3. The stem cell nature of Wt MDSC and response to paracrine effects is evidenced by their conversion into SMC when

Mercury and other trace elements in Ohio River fish collected near coal-fired power plants: Interspecific patterns and consideration of consumption risks.

PubMed

Reash, Robin J; Brown, Lauren; Merritt, Karen

2015-07-01

Many coal-fired electric generating facilities in the United States are discharging higher loads of Hg, Se, and other chemicals to receiving streams due to the installation of flue gas desulfurization (FGD) air pollution control units. There are regulatory concerns about the potential increased uptake of these bioaccumulative trace elements into food webs. We evaluated the concentrations of As, total Hg (THg), methylmercury (MeHg), and Se in Ohio River fish collected proximal to coal-fired power plants, of which 75% operate FGD systems. Fillet samples (n = 50) from 6 fish species representing 3 trophic levels were analyzed. Geometric mean fillet concentrations of THg (wet wt), MeHg (wet wt), and Se (dry wt) in 3 species were 0.136, 0.1181, and 3.19 mg/kg (sauger); 0.123, 0.1013, and 1.56 mg/kg (channel catfish); and 0.127, 0.0914, and 3.30 mg/kg (hybrid striped bass). For all species analyzed, only 3 fillet samples (6% of total) had MeHg concentrations that exceeded the US Environmental Protection Agency (USEPA) human health criterion (0.3 mg/kg wet wt); all of these were freshwater drum aged ≥ 19 y. None of the samples analyzed exceeded the USEPA proposed muscle and whole body Se thresholds for protection against reproductive effects in freshwater fish. All but 8 fillet samples had a total As concentration less than 1.0 mg/kg dry wt. Mean Se health benefit values (HBVSe ) for all species were ≥ 4, indicating that potential Hg-related health risks associated with consumption of Ohio River fish are likely to be offset by adequate Se concentrations. Overall, we observed no measurable evidence of enhanced trace element bioaccumulation associated with proximity to power plant FGD facilities, however, some enhanced bioaccumulation could have occurred in the wastewater mixing zones. Furthermore, available evidence indicates that, due to hydraulic and physical factors, the main stem Ohio River appears to have low net Hg methylation potential. © 2015 SETAC.

In vitro blood and fibroblast responses to BisGMA-TEGDMA/bioactive glass composite implants.

PubMed

Abdulmajeed, Aous A; Kokkari, Anne K; Käpylä, Jarmo; Massera, Jonathan; Hupa, Leena; Vallittu, Pekka K; Närhi, Timo O

2014-01-01

This in vitro study was designed to evaluate both blood and human gingival fibroblast responses to bisphenol A-glycidyl methacrylate-triethyleneglycol dimethacrylate (BisGMA-TEGDMA)/bioactive glass (BAG) composite, aimed to be used as composite implant abutment surface modifier. Three different types of substrates were investigated: (a) plain polymer (BisGMA 50 wt%-TEGDMA 50 wt%), (b) BAG-composite (50 wt% polymer + 50 wt% fraction of BAG-particles, <50 μm), and (c) plain BAG plates (100 wt% BAG). The blood response, including the blood-clotting ability and platelet adhesion morphology were evaluated. Human gingival fibroblasts were plated and cultured on the experimental substrates for up to 10 days, then the cell proliferation rate was assessed using AlamarBlue assay™. The BAG-composite and plain BAG substrates had a shorter clotting time than plain polymer substrates. Platelet activation and aggregation were most extensive, qualitatively, on BAG-composite. Analysis of the normalized cell proliferation rate on the different surfaces showed some variations throughout the experiment, however, by day 10 the BAG-composite substrate showed the highest (P < 0.001) cell proliferation rate. In conclusion, the presence of exposed BAG-particles enhances fibroblast and blood responses on composite surfaces in vitro.

Flexible bactericidal graphene oxide-chitosan layers for stem cell proliferation

NASA Astrophysics Data System (ADS)

Mazaheri, M.; Akhavan, O.; Simchi, A.

2014-05-01

Graphene oxide (GO)-chitosan composite layers with stacked layer structures were synthesized using chemically exfoliated GO sheets (with lateral dimensions of ˜1 μm and thickness of ˜1 nm), and applied as antibacterial and flexible nanostructured templates for stem cell proliferation. By increasing the GO content from zero to 6 wt%, the strength and elastic modulus of the layers increased ˜80% and 45%, respectively. Similar to the chitosan layer, the GO-chitosan composite layers showed significant antibacterial activity (>77% inactivation after only 3 h) against Staphylococcus aureus bacteria. Surface density of the actin cytoskeleton fibers of human mesenchymal stem cells (hMSCs) cultured on the chitosan and GO(1.5 wt%)-chitosan composite layers was found nearly the same, while it significantly decreased by increasing the GO content to 3 and 6 wt%. Our results indicated that although a high concentration of GO in the chitosan layer (here, 6 wt%) could decelerate the proliferation of the hMSCs on the flexible layer, a low concentration of GO (i.e., 1.5 wt%) not only resulted in biocompatibility but also kept the mechanical flexibility of the self-sterilized layers for high proliferation of hMSCs.

Ternary Blend Composed of Two Organic Donors and One Acceptor for Active Layer of High-Performance Organic Solar Cells.

PubMed

Lee, Jong Won; Choi, Yoon Suk; Ahn, Hyungju; Jo, Won Ho

2016-05-04

Ternary blends composed of two donor absorbers with complementary absorptions provide an opportunity to enhance the short-circuit current and thus the power conversion efficiency (PCE) of organic solar cells. In addition to complementary absorption of two donors, ternary blends may exhibit favorable morphology for high-performance solar cells when one chooses properly the donor pair. For this purpose, we develop a ternary blend with two donors (diketopyrrolopyrrole-based polymer (PTDPP2T) and small molecule ((TDPP)2Ph)) and one acceptor (PC71BM). The solar cell made of a ternary blend with 10 wt % (TDPP)2Ph exhibits higher PCE of 7.49% as compared with the solar cells with binary blends, PTDPP2T:PC71BM (6.58%) and (TDPP)2Ph:PC71BM (3.21%). The higher PCE of the ternary blend solar cell is attributed mainly to complementary absorption of two donors. However, a further increase in (TDPP)2Ph content in the ternary blend (>10 wt %) decreases the PCE. The ternary blend with 10 wt % (TDPP)2Ph exhibits well-developed morphology with narrow-sized fibrils while the blend with 15 wt % (TDPP)2Ph shows phase separation with large-sized domains, demonstrating that the phase morphology and compatibility of ternary blend are important factors to achieve a high-performance solar cell made of ternary blends.

Simulation of Zr content in TiZrCuNi brazing filler metal for Ti6Al4V alloy

NASA Astrophysics Data System (ADS)

Yue, Xishan; Xie, Zonghong; Jing, Yongjuan

2017-07-01

To optimize the Zr content in Ti-based filler metal, the covalent electron on the nearest atoms bond in unit cell ( n A u-v ) with Ti-based BCC structure was calculated, in which the brazing temperature was considered due to its influence on the lattice parameter. Based on EET theory (The Empirical Electron Theory for solid and molecules), n_{{A}}^{{u - v}} represents the strength of the unit cell with defined element composition and structure, which reflects the effect from solid solution strengthening on the strength of the unit cell. For Ti-Zr-15Cu-10Ni wt% filler metal, it kept constant as 0.3476 with Zr as 37.5˜45 wt% and decreased to 0.333 with Zr decreasing from 37.5 to 25 wt%. Finally, it increased up to 0.3406 with Zr as 2˜10 wt%. Thus, Ti-based filler metal with Zr content being 2˜10 wt% is suggested based on the simulation results. Moreover, the calculated covalent electron of n A u-v showed good agreement with the hardness of the joint by filler 37.5Zr and 10Zr. The composition of Ti-10Zr-15Cu-10Ni wt% was verified in this study with higher tensile strength of the brazing joint and uniform microstructure of the interface.

In vitro cytotoxicity and in vivo osseointergration properties of compression-molded HDPE-HA-Al2O3 hybrid biocomposites.

PubMed

Tripathi, Garima; Gough, Julie E; Dinda, Amit; Basu, Bikramjit

2013-06-01

The aim of this study was to investigate the in vivo biocompatibility in terms of healing of long segmental bone defect in rabbit model as well as in vitro cytotoxicity of eluates of compression-molded High density polyethylene (HDPE)-hydroxyapatite (HA)-aluminum oxide (Al2O3) composite-based implant material. Based on the physical property in terms of modulus and strength properties, as reported in our recent publication, HDPE-40 wt % HA and HDPE-20 wt % HA-20 wt % Al2O3 hybrid composites were used for biocompatibility assessment. Osteoblasts cells were cultured in conditioned media, which contains varying amount of composite eluate (0.01, 0.1, and 1.0 wt %). In vitro, the eluates did not exhibit any significant negative impact on proliferation, mineralization or on morphology of human osteoblast cells. In vivo, the histological assessment revealed neobone formation at the bone/implant interface, characterized by the presence of osteoid and osteoblasts. The observation of osteoclastic activity indicates the process of bone remodeling. No inflammation to any noticeable extent was observed at the implantation site. Overall, the combination of in vitro and in vivo results are suggestive of potential biomedical application of compression-molded HDPE- 20 wt % HA- 20 wt % Al2O3 composites to heal long segmental bone defects without causing any toxicity of bone cells. Copyright © 2012 Wiley Periodicals, Inc.

GENDER DIFFERENCES IN INJURY INDUCED MESENCHYMAL STEM CELL APOPTOSIS, EXPRESSION OF VEGF, TNF, AND IL-6 AND ABROGATION VIA TNFR1 ABLATION

PubMed Central

Crisostomo, Paul R.; Wang, Meijing; Herring, Christine M.; Markel, Troy A.; Meldrum, Kirstan K.; Lillemoe, Keith D.; Meldrum, Daniel R.

2007-01-01

Concomitant pro- and anti-inflammatory properties of bone marrow stem cells (BMSC) may be an important aspect of their ability to heal injured tissue. However, very few studies have examined whether gender differences exist in BMSC function. Indeed, it remains unknown whether gender differences exist in BMSC function and ability to resist apoptosis, and if so, whether TNF receptor 1 (TNFR1) plays a role in these differences. We hypothesized that TNFR1 ablation equalizes gender differences in bone marrow mesenchymal stem cell (MSC) apoptosis, as well as expression of vascular endothelial growth factor (VEGF), TNF, and interleukin (IL)-6. Mouse MSCs from male wildtype (WT), female WT, male TNFR1 knockouts (TNFR1KO), and female TNFR1KO were stressed by endotoxin 200 ng/ml or 1 hr hypoxia. MSC activation was determined by measuring VEGF, TNF, and IL-6 production (ELISA). Differences considered significant if p<0.05. LPS and hypoxia resulted in significant activation in all experimental groups compared to controls. Male WT demonstrated significantly greater TNF and IL-6 and significantly less VEGF release than female WT MSCs. However, release of TNF, IL-6, and VEGF in male TNFR1 knockouts differed from male WT, but was not different from female WT MSCs. Similarly apoptosis in hypoxic male TNFRIKO differed from male WT, but it was not different from apoptosis from WT female. Female WT did not differ in TNF, IL-6, and VEGF release compared to female TNFR1KO. Gender differences exist in injury induced BMSC VEGF, TNF, and IL-6 expression. TNFR1 may autoregulate VEGF, TNF, and IL-6 expression in males more than females. MSCs are novel therapeutic agents for organ protection, but further study of the disparate expression of VEGF, TNF, and IL-6 in males and females as well as the role of TNFR1 in these gender differences is necessary to maximize this protection. PMID:17070836

Podocyte-specific deletion of Rac1 leads to aggravation of renal injury in STZ-induced diabetic mice

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ishizaka, Masanori; Gohda, Tomohito, E-mail: goda@juntendo.ac.jp; Takagi, Miyuki

Rac1, a GTPase of the Rho subfamily, has a crucial role in cytoskeletal architecture, as well as the regulation of cell migration and growth. However, renal injury in mice with podocyte-specific deletion of Rac1 has yet to be elucidated fully due to conflicting findings. Herein, we identified a possible role for Rac1 in podocytes of streptozotocin- (STZ) induced diabetic mice. The urinary albumin/creatinine ratio (ACR) in the knockout (KO) group was significantly higher than that in the wild type (WT) group at any week of age. A more marked ACR increase was observed in STZ/KO group than STZ/WT group, althoughmore » ACR did increase with weeks of age in both diabetic groups. The kidney sections from diabetic mice revealed a glomerular hypertrophy with mesangial expansion, but there was no appreciable difference in glomerular findings under a light microscope between STZ/WT and STZ/KO mice. However, an electron microscopy analysis revealed that regardless of the presence or absence of diabetes, both KO (KO and STZ/KO) groups had a higher rate of foot process effacement compared with both WT (WT and STZ/WT) groups. The expression levels of the slit diaphragm protein, podocin, was reduced with the induction of diabetes, and the levels in the STZ/KO group experienced a further reduction compared with the STZ/WT group. The number of WT1-positive cells in the STZ/KO group was more significantly decreased than that in the other three groups. In contrast, the numbers of cleaved caspase 3- and TUNEL-positive cells in the glomeruli of the STZ/KO group were more increased than those in the STZ/WT group. Thus, this study provides evidence that podocyte-specific deletion of Rac1 results in morphological alteration in podocytes, and that the induction of apoptosis or decreased expression of the slit diaphragm proteins by hyperglycemic stimuli are associated with the progression of diabetic nephropathy.« less

Keratins Regulate p38MAPK-Dependent Desmoglein Binding Properties in Pemphigus

PubMed Central

Vielmuth, Franziska; Walter, Elias; Fuchs, Michael; Radeva, Mariya Y.; Buechau, Fanny; Magin, Thomas M.; Spindler, Volker; Waschke, Jens

2018-01-01

Keratins are crucial for the anchorage of desmosomes. Severe alterations of keratin organization and detachment of filaments from the desmosomal plaque occur in the autoimmune dermatoses pemphigus vulgaris and pemphigus foliaceus (PF), which are mainly caused by autoantibodies against desmoglein (Dsg) 1 and 3. Keratin alterations are a structural hallmark in pemphigus pathogenesis and correlate with loss of intercellular adhesion. However, the significance for autoantibody-induced loss of intercellular adhesion is largely unknown. In wild-type (wt) murine keratinocytes, pemphigus autoantibodies induced keratin filament retraction. Under the same conditions, we used murine keratinocytes lacking all keratin filaments (KtyII k.o.) as a model system to dissect the role of keratins in pemphigus. KtyII k.o. cells show compromised intercellular adhesion without antibody (Ab) treatment, which was not impaired further by pathogenic pemphigus autoantibodies. Nevertheless, direct activation of p38MAPK via anisomycin further decreased intercellular adhesion indicating that cell cohesion was not completely abrogated in the absence of keratins. Direct inhibition of Dsg3, but not of Dsg1, interaction via pathogenic autoantibodies as revealed by atomic force microscopy was detectable in both cell lines demonstrating that keratins are not required for this phenomenon. However, PF-IgG shifted Dsg1-binding events from cell borders toward the free cell surface in wt cells. This led to a distribution pattern of Dsg1-binding events similar to KtyII k.o. cells under resting conditions. In keratin-deficient keratinocytes, PF-IgG impaired Dsg1-binding strength, which was not different from wt cells under resting conditions. In addition, pathogenic autoantibodies were capable of activating p38MAPK in both KtyII wt and k.o. cells, the latter of which already displayed robust p38MAPK activation under resting conditions. Since inhibition of p38MAPK blocked autoantibody-induced loss of intercellular adhesion in wt cells and restored baseline cell cohesion in keratin-deficient cells, we conclude that p38MAPK signaling is (i) critical for regulation of cell adhesion, (ii) regulated by keratins, and (iii) targets both keratin-dependent and -independent mechanisms. PMID:29616033

Effect of the relationship between particle size, inter-particle distance, and metal loading of carbon supported fuel cell catalysts on their catalytic activity

NASA Astrophysics Data System (ADS)

Corradini, Patricia Gon; Pires, Felipe I.; Paganin, Valdecir A.; Perez, Joelma; Antolini, Ermete

2012-09-01

The effect of the relationship between particle size ( d), inter-particle distance ( x i ), and metal loading ( y) of carbon supported fuel cell Pt or PtRu catalysts on their catalytic activity, based on the optimum d (2.5-3 nm) and x i / d (>5) values, was evaluated. It was found that for y < 30 wt%, the optimum values of both d and x i / d can be always obtained. For y ≥ 30 wt%, instead, the positive effect of a thinner catalyst layer of the fuel cell electrode than that using catalysts with y < 30 wt% is concomitant to a decrease of the effective catalyst surface area due to an increase of d and/or a decrease of x i / d compared to their optimum values, with in turns gives rise to a decrease in the catalytic activity. The effect of the x i / d ratio has been successfully verified by experimental results on ethanol oxidation on PtRu/C catalysts with same particle size and same degree of alloying but different metal loading. Tests in direct ethanol fuel cells showed that, compared to 20 wt% PtRu/C, the negative effect of the lower x i / d on the catalytic activity of 30 and 40 wt% PtRu/C catalysts was superior to the positive effect of the thinner catalyst layer.

p53 is important for the anti-proliferative effect of ibuprofen in colon carcinoma cells

DOE Office of Scientific and Technical Information (OSTI.GOV)

Janssen, Astrid; Schiffmann, Susanne; Birod, Kerstin

2008-01-25

S-ibuprofen which inhibits the cyclooxygenase-1/-2 and R-ibuprofen which shows no COX-inhibition at therapeutic concentrations have anti-carcinogenic effects in human colon cancer cells; however, the molecular mechanisms for these effects are still unknown. Using HCT-116 colon carcinoma cell lines, expressing either the wild-type form of p53 (HCT-116 p53{sup wt}) or being p(HCT-116 p53{sup -/-}), we demonstrated that both induction of a cell cycle block and apoptosis after S- and R-ibuprofen treatment is in part dependent on p53. Also in the in vivo nude mice model HCT-116 p53{sup -/-} xenografts were less sensitive for S- and R-ibuprofen treatment than HCT-116 p53{sup wt}more » cells. Furthermore, results indicate that induction of apoptosis in HCT-116 p53{sup wt} cells after ibuprofen treatment is in part dependent on a signalling pathway including the neutrophin receptor p75{sup NTR}, p53 and Bax.« less

Overexpression or absence of calretinin in mouse primary mesothelial cells inversely affects proliferation and cell migration.

PubMed

Blum, Walter; Pecze, László; Felley-Bosco, Emanuela; Schwaller, Beat

2015-12-22

The Ca(2+)-binding protein calretinin is currently used as a positive marker for identifying epithelioid malignant mesothelioma (MM) and reactive mesothelium, but calretinin's likely role in mesotheliomagenesis remains unclear. Calretinin protects immortalized mesothelial cells in vitro from asbestos-induced cytotoxicity and thus might be implicated in mesothelioma formation. To further investigate calretinin's putative role in the early steps of MM generation, primary mesothelial cells from calretinin knockout (CR-/-) and wildtype (WT) mice were compared. Primary mouse mesothelial cells from WT and CR-/- mice were investigated with respect to morphology, marker proteins, proliferation, cell cycle parameters and mobility in vitro. Overexpression of calretinin or a nuclear-targeted variant was achieved by a lentiviral expression system. CR-/- mice have a normal mesothelium and no striking morphological abnormalities compared to WT animals were noted. Primary mouse mesothelial cells from both genotypes show a typical "cobblestone-like" morphology and express mesothelial markers including mesothelin. In cells from CR-/- mice in vitro, we observed more giant cells and a significantly decreased proliferation rate. Up-regulation of calretinin in mesothelial cells of both genotypes increases the proliferation rate and induces a cobblestone-like epithelial morphology. The length of the S/G2/M phase is unchanged, however the G1 phase is clearly prolonged in CR-/- cells. They are also much slower to close a scratch in a confluent cell layer (2D-wound assay). In addition to a change in cell morphology, an increase in proliferation and mobility is observed, if calretinin overexpression is targeted to the nucleus. Thus, both calretinin and nuclear-targeted calretinin increase mesothelial cell proliferation and consequently, speed up the scratch-closure time. The increased rate of scratch closure in WT cells is the result of two processes: an increased proliferation rate and augmented cell mobility of the border cells migrating towards the empty space. We hypothesize that the differences in proliferation and mobility between WT and CR-/- mesothelial cells are the likely result from differences in their developmental trajectories. The mechanistic understanding of the function of calretinin and its putative implication in signaling pathways in normal mesothelial cells may help understanding its role during the processes that lead to mesothelioma formation and could possibly open new avenues for mesothelioma therapy, either by directly targeting calretinin expression or indirectly by targeting calretinin-mediated downstream signaling.

Warthin-like Mucoepidermoid Carcinoma: A Combined Study of Fluorescence In Situ Hybridization and Whole-slide Imaging.

PubMed

Ishibashi, Kenichiro; Ito, Yohei; Masaki, Ayako; Fujii, Kana; Beppu, Shintaro; Sakakibara, Takeo; Takino, Hisashi; Takase, Hiroshi; Ijichi, Kei; Shimozato, Kazuo; Inagaki, Hiroshi

2015-11-01

There has been some debate as to whether a subset of metaplastic Warthin tumors (mWTs) harbor the mucoepidermoid carcinoma (MEC)-associated CRTC1-MAML2 fusion. We analyzed 15 tumors originally diagnosed as mWT (mWT-like tumors), 2 of which had concurrent MECs. We looked for the CRTC1/3-MAML2 fusion transcripts and performed immunohistochemistry for p63 and fluorescence in situ hybridization (FISH) for the MAML2 split. To localize MAML2 split-positive cells at the cellular level, whole tumor tissue sections were digitalized (whole-slide imaging [WSI]). The CRTC1-MAML2, but not CRTC3-MAML2 was detected in 5/15 mWT-like tumors. FISH-WSI results showed that all epithelial cells harbored the MAML2 split in fusion-positive mWT-like tumors and were totally negative in fusion-negative mWT-like tumors. A review of the hematoxylin and eosin-stained slides showed that morphology of the "metaplastic" epithelium was virtually indistinguishable between fusion-positive and fusion-negative tumors. However, oncocytic bilayered tumor epithelium, characteristic to typical WT, was always found somewhere in the fusion-negative tumors but not in the fusion-positive tumors. This distinguishing histologic finding enabled 5 pathologists to easily differentiate the 2 tumor groups with 100% accuracy. The age and sex distribution of fusion-positive mWT-like tumor cases was similar to that of fusion-positive MEC cases and significantly different from those of fusion-negative mWT-like tumor and typical WT cases. In addition, only fusion-positive mWT-like tumors possessed concurrent low-grade MECs. In conclusion, a subset of mWT-like tumors were positive for the CRTC1-MAML2 fusion and had many features that are more in accord with MEC than with WT. The term Warthin-like MEC should be considered for fusion-positive mWT-like tumors.

Atomic structures of fibrillar segments of hIAPP suggest tightly mated β-sheets are important for cytotoxicity

DOE Office of Scientific and Technical Information (OSTI.GOV)

Krotee, Pascal; Rodriguez, Jose A.; Sawaya, Michael R.

2017-01-03

hIAPP fibrils are associated with Type-II Diabetes, but the link of hIAPP structure to islet cell death remains elusive. Here we observe that hIAPP fibrils are cytotoxic to cultured pancreatic β-cells, leading us to determine the structure and cytotoxicity of protein segments composing the amyloid spine of hIAPP. Using the cryoEM method MicroED, we discover that one segment, 19–29 S20G, forms pairs of β-sheets mated by a dry interface that share structural features with and are similarly cytotoxic to full-length hIAPP fibrils. In contrast, a second segment, 15–25 WT, forms non-toxic labile β-sheets. These segments possess different structures and cytotoxicmore » effects, however, both can seed full-length hIAPP, and cause hIAPP to take on the cytotoxic and structural features of that segment. These results suggest that protein segment structures represent polymorphs of their parent protein and that segment 19–29 S20G may serve as a model for the toxic spine of hIAPP.« less

Tim-3 directly enhances CD8 T cell responses to acute Listeria monocytogenes infection

PubMed Central

Gorman, Jacob V.; Starbeck-Miller, Gabriel; Pham, Nhat-Long L.; Traver, Geri L.; Rothman, Paul B.; Harty, John T.; Colgan, John D.

2014-01-01

Tim-3 is a surface molecule expressed throughout the immune system that can mediate both stimulatory and inhibitory effects. Previous studies have provided evidence that Tim-3 functions to enforce CD8 T cell exhaustion, a dysfunctional state associated with chronic stimulation. In contrast, the role of Tim-3 in the regulation of CD8 T cell responses to acute and transient stimulation remains undefined. To address this knowledge gap, we examined how Tim-3 affects CD8 T cell responses to acute Listeria monocytogenes (LM) infection. Analysis of wild-type (WT) mice infected with LM revealed that Tim-3 was transiently expressed by activated CD8 T cells and was associated primarily with acquisition of an effector phenotype. Comparison of responses to LM by WT and Tim-3 KO mice showed that the absence of Tim-3 significantly reduced the magnitudes of both primary and secondary CD8 T cell responses, which correlated with decreased IFN-γ production and degranulation by Tim-3 KO cells stimulated with peptide antigen ex vivo. To address the T cell-intrinsic role of Tim-3, we analyzed responses to LM infection by WT and Tim-3 KO TCR-transgenic CD8 T cells following adoptive transfer into a shared WT host. In this setting, the accumulation of CD8 T cells and the generation of cytokine-producing cells were significantly reduced by the lack of Tim-3, demonstrating that this molecule has a direct effect on CD8 T cell function. Combined, our results suggest that Tim-3 can mediate a stimulatory effect on CD8 T cell responses to an acute infection. PMID:24567532

Fibrin-polyethylene oxide interpenetrating polymer networks: new self-supported biomaterials combining the properties of both protein gel and synthetic polymer.

PubMed

Akpalo, E; Bidault, L; Boissière, M; Vancaeyzeele, C; Fichet, O; Larreta-Garde, V

2011-06-01

Interpenetrating polymer network (IPN) architectures were conceived to improve the mechanical properties of a fibrin gel. Conditions allowing an enzymatic reaction to create one of the two networks in IPN architecture were included in the synthesis pathway. Two IPN series were carried out, starting from two polyethylene oxide (PEO) network precursors leading to different cross-linking densities of the PEO phase. The fibrin concentration varied from 5 to 20 wt.% in each series. The behavior of these materials during dehydration/hydration cycles was also studied. The mechanical properties of the resulting IPN were characterized in the wet and dry states. These self-supported biomaterials combine the properties of both a protein gel and a synthetic polymer. Finally, cells were grown on PEO/fibrin IPN, indicating that they are non-cytotoxic. Copyright © 2011 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Polyhydroxybutyrate accumulation by a Serratia sp.

PubMed

Lugg, Harriet; Sammons, Rachel L; Marquis, Peter M; Hewitt, Christopher J; Yong, Ping; Paterson-Beedle, Marion; Redwood, Mark D; Stamboulis, Artemis; Kashani, Mitra; Jenkins, Mike; Macaskie, Lynne E

2008-03-01

A strain of Serratia sp. showed intracellular electron-transparent inclusion bodies when incubated in the presence of citrate and glycerol 2-phosphate without nitrogen source following pre-growth under carbon-limitation in continuous culture. About 1.3 mmol citrate were consumed per 450 mg biomass, giving a calculated yield of maximally 55% of stored material per g of biomass dry wt. The inclusion bodies were stained with Sudan Black and Nile Red (NR), suggesting a lipid material, which was confirmed as polyhydroxybutyrate (PHB) by analysis of molecular fragments by GC and by FTIR spectroscopy of isolated bio-PHB in comparison with reference material. Multi-parameter flow cytometry in conjunction with NR fluorescence, and electron microscopy, showed that not all cells contained heavy PHB bodies, suggesting the potential for increasing the overall yield. The economic attractiveness is enhanced by the co-production of nanoscale hydroxyapatite (HA), a possible high-value precursor for bone replacement materials.

Expression of lycopene biosynthesis genes fused in line with Shine-Dalgarno sequences improves the stress-tolerance of Lactococcus lactis.

PubMed

Dong, Xiangrong; Wang, Yanping; Yang, Fengyuan; Zhao, Shanshan; Tian, Bing; Li, Tao

2017-01-01

Lycopene biosynthetic genes from Deinococcus radiodurans were co-expressed in Lactococcus lactis to produce lycopene and improve its tolerance to stress. Lycopene-related genes from D. radiodurans, DR1395 (crtE), DR0862 (crtB), and DR0861 (crtI), were fused in line with S hine-Dalgarno (SD) sequences and co-expressed in L. lactis. The recombinant strain produced 0.36 mg lycopene g -1  dry cell wt after 48 h fermentation. The survival rate to UV irradiation of the recombinant strain was higher than that of the non-transformed strain. The L. lactis with co-expressed genes responsible for lycopene biosynthesis from D. radiodurans produced lycopene and exhibited increased resistance to UV stress, suggesting that the recombinant strain has important application potential in food industry.

Assessing storage of stability and mercury reduction of freeze-dried Pseudomonas putida within different types of lyoprotectant

NASA Astrophysics Data System (ADS)

Azoddein, Abdul Aziz Mohd; Nuratri, Yana; Azli, Faten Ahada Mohd; Bustary, Ahmad Bazli

2017-12-01

Pseudomonas putida is a potential strain in biological treatment to remove mercury contained in the effluent of petrochemical industry due to its mercury reductase enzyme that able to reduce ionic mercury to elementary mercury. Freeze-dried P. putida allows easy, inexpensive shipping, handling and high stability of the product. This study was aimed to freeze dry P. putida cells with addition of lyoprotectant. Lyoprotectant was added into the cells suspension prior to freezing. Dried P. putida obtained was then mixed with synthetic mercury. Viability of recovery P. putida after freeze dry was significantly influenced by the type of lyoprotectant. Among the lyoprotectants, tween 80/ sucrose was found to be the best lyoprotectant. Sucrose was able to recover more than 78% (6.2E+09 CFU/ml) of the original cells (7.90E+09CFU/ml) after freeze dry and able to retain 5.40E+05 viable cells after 4 weeks storage at 4 °C without vacuum. Polyethylene glycol (PEG) pre-treated freeze dried cells and broth pre-treated freeze dried cells after the freeze-dry process recovered more than 64% (5.0 E+09CFU/ml) and >0.1% (5.60E+07CFU/ml). Freeze-dried P. putida cells in PEG and broth cannot survive after 4 weeks storage. Freeze dry also does not really change the pattern of growth P. putida but extension of lag time was found 1 hour after 3 weeks of storage. Additional time was required for freeze-dried P. putida cells to recover before introducing freeze-dried cells to more complicated condition such as mercury solution. The maximum mercury reduction of PEG pre-treated freeze-dried cells after freeze dry and after storage of 3 weeks was 17.91 %. The maximum of mercury reduction of tween 80/sucrose pre-treated freeze-dried cells after freeze dry and after storage 3 weeks was 25.03%. Freeze dried P. putida was found to have lower mercury reduction compare to the fresh P. putida that has been grown in agar. Result from this study may be beneficial and useful as initial reference before commercialized freeze-dried P. putida.

Cyclooxygenase-2 deficiency impairs muscle-derived stem cell-mediated bone regeneration via cellular autonomous and non-autonomous mechanisms.

PubMed

Gao, Xueqin; Usas, Arvydas; Lu, Aiping; Kozemchak, Adam; Tang, Ying; Poddar, Minakshi; Sun, Xuying; Cummins, James H; Huard, Johnny

2016-08-01

This study investigated the role of cyclooxygenase-2 (COX-2) expression by donor and host cells in muscle-derived stem cell (MDSC)-mediated bone regeneration utilizing a critical size calvarial defect model. We found that BMP4/green fluorescent protein (GFP)-transduced MDSCs formed significantly less bone in COX-2 knock-out (Cox-2KO) than in COX-2 wild-type (WT) mice. BMP4/GFP-transduced Cox-2KO MDSCs also formed significantly less bone than transduced WT MDSCs when transplanted into calvarial defects created in CD-1 nude mice. The impaired bone regeneration in the Cox-2KO MDSCBMP4/GFP group is associated with downregulation of BMP4-pSMAD1/5 signaling, decreased osteogenic differentiation and lowered proliferation capacity after transplantation, compared with WT MDSCBMP4/GFP cells. The Cox-2KO MDSCBMP4/GFP group demonstrated a reduction in cell survival and direct osteogenic differentiation in vitro These effects were mediated in part by the downregulation of Igf1 and Igf2. In addition, the Cox-2KO MDSCBMP4/GFP cells recruited fewer macrophages than the WT MDSC/BMP4/GFP cells in the early phase after injury. We concluded that the bone regeneration capacity of Cox-2KO MDSCs was impaired because of a reduction in cell proliferation and survival capacities, reduction in osteogenic differentiation and a decrease in the ability of the cells to recruit host cells to the injury site. © The Author 2016. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Electrochemical impedance spectroscopy study of carbon electrodes prepared from date pits and fibers of oil palm empty fruit bunches

NASA Astrophysics Data System (ADS)

Hamdan, E.; Deraman, M.; Suleman, M.; Nor, N. S. M.; Basri, N. H.; Hanappi, M. F. Y. M.; Sazali, N. E. S.; Tajuddin, N. S. M.; Omar, R.; Othman, M. A. R.; Shamsudin, S. A.

2016-11-01

In this study, we produced pre-carbonized date pits (PDP) and self-adhesive carbon grains (SACGs) from oil palm empty fruit bunches (EFB) by a low temperature (200°C for DP and 280°C for SACGs, respectively) carbonization method followed by KOH treatment to obtain KOH treated PDP (T-PDP) and KOH treated SACGs (T-SACGs). Four sets of green monolith (GMs) denoted as GM-A, GM-B, GM-C and GM-D were prepared respectively from SACGs (100 wt. %), mixture of PDP and SACGs (50:50 wt. %), T-SACGs (100 wt. %), and mixture of T-SACGs and T-PDP (50:50 wt. %), respectively. From these GMs the respective activated carbon monolith (ACMs) electrodes namely ACM-A, ACM-B, ACM-C and ACM-D were prepared via carbonization (N2 carbonization) and activation (CO2 environment). These ACMs electrodes were used to fabricate the corresponding EDLC cells: Cell-A, Cell-B, Cell-C and Cell-D, respectively. The electrochemical impedance spectroscopy tests conducted on the cells found that the Cell-D showed the maximum value of specific capacitance, Csp (˜ 135 F g-1) whereas the Cell-A showed the minimum values of ESR and characteristic response time, respectively, ˜ 2.14 Ω and ˜ 46 s. Therefore, it can be concluded that the KOH treatment can improve the capacitance but caused the increase in the ESR and response time.

Isolation and characterization of dental epithelial cells derived from amelogenesis imperfecta rat.

PubMed

Adiningrat, A; Tanimura, A; Miyoshi, K; Hagita, H; Yanuaryska, R D; Arinawati, D Y; Horiguchi, T; Noma, T

2016-03-01

Disruption of the third zinc finger domain of specificity protein 6 (SP6) presents an enamel-specific defect in a rat model of amelogenesis imperfecta (AMI rats). To understand the molecular basis of amelogenesis imperfecta caused by the Sp6 mutation, we established and characterized AMI-derived rat dental epithelial (ARE) cells. ARE cell clones were isolated from the mandibular incisors of AMI rats, and amelogenesis-related gene expression was analyzed by reverse transcription polymerase chain reaction (RT-PCR). Localization of wild-type SP6 (SP6WT) and mutant-type SP6 (SP6AMI) was analyzed by immunocytochemistry. SP6 transcriptional activity was monitored by rho-associated protein kinase 1 (Rock1) promoter activity with its specific binding to the promoter region in dental (G5 and ARE) and non-dental (COS-7) epithelial cells. Isolated ARE cells were varied in morphology and gene expression. Both SP6WT and SP6AMI were mainly detected in nuclei. The promoter analysis revealed that SP6WT and SP6AMI enhanced Rock1 promoter activity in G5 cells but that enhancement by SP6AMI was weaker, whereas no enhancement was observed in the ARE and COS-7 cells, even though SP6WT and SP6AMI bound to the promoter in all instances. ARE cell clones can provide a useful in vitro model to study the mechanism of SP6-mediated amelogenesis imperfecta. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Decoupling of the PI3K pathway via mutation necessitates combinatorial treatment in HER2+ breast cancer

DOE PAGES

Korkola, James E.; Collisson, Eric A.; Heiser, Laura; ...

2015-07-16

We report here on experimental and theoretical efforts to determine how best to combine drugs that inhibit HER2 and AKT in HER2 + breast cancers. We accomplished this by measuring cellular and molecular responses to lapatinib and the AKT inhibitors (AKT i) GSK690693 and GSK2141795 in a panel of 22 HER2 + breast cancer cell lines carrying wild type or mutant PIK3CA. We observed that combinations of lapatinib plus AKT i were synergistic in HER2 /PIK3CA mut cell lines but not in HER2 +/PIK3CA wt cell lines. We measured changes in phospho-protein levels in 15 cell lines after treatment withmore » lapatinib, AKT i or lapatinib + AKT i to shed light on the underlying signaling dynamics. This revealed that p-S6RP levels were less well attenuated by lapatinib in HER2 +/PIK3CA mut cells compared to HER2 +/PIK3CA wt cells and that lapatinib + AKT i reduced p-S6RP levels to those achieved in HER2 +/PIK3CA wt cells with lapatinib alone. We also found that that compensatory up-regulation of p-HER3 and p-HER2 is blunted in PIK3CA mut cells following lapatinib + AKT i treatment. Responses of HER2 + SKBR3 cells transfected with lentiviruses carrying control or PIK3CA mut sequences were similar to those observed in HER2 +/PIK3CA mut cell lines but not in HER2 +/PIK3CA wt cell lines. We used a nonlinear ordinary differential equation model to support the idea that PIK3CA mutations act as downstream activators of AKT that blunt lapatinib inhibition of downstream AKT signaling and that the effects of PIK3CA mutations can be countered by combining lapatinib with an AKT i. This combination does not confer substantial benefit beyond lapatinib in HER2 +/PIK3CA wt cells.« less

NOS2 deficiency has no influence on the radiosensitivity of the hematopoietic system.

PubMed

Li, Chengcheng; Luo, Yi; Shao, Lijian; Meng, Aimin; Zhou, Daohong

2018-01-01

Previous studies have shown that inhibition of inducible NO synthase (NOS2 or iNOS) with an inhibitor can selectively protect several normal tissues against radiation during radiotherapy. However, the role of NOS2 in ionizing radiation (IR)-induced bone marrow (BM) suppression is unknown and thus was investigated in the present study using NOS2 - / - and wild-type mice 14 days after they were exposed to a sublethal dose of total body irradiation (TBI). The effects of different doses of IR (1, 2 and 4 Gy) on the apoptosis and colony-forming ability of bone marrow cells from wild-type (WT) and NOS2 - / - mice were investigated in vitro. In addition, we exposed NOS2 - / - mice and WT mice to 6-Gy TBI or sham irradiation. They were euthanized 14 days after TBI for analysis of peripheral blood cell counts and bone marrow cellularity. Colony-forming unit-granulocyte and macrophage, burst-forming unit-erythroid and CFU-granulocyte, erythroid, macrophage in bone marrow cells from the mice were determined to evaluate the function of hematopoietic progenitor cells (HPCs), and the ability of hematopoietic stem cells (HSCs) to self-renew was analysed by the cobblestone area forming cell assay. The cell cycling of HPCs and HSCs were measured by flow cytometry. Exposure to 2 and 4 Gy IR induced bone marrow cell apoptosis and inhibited the proliferation of HPCs in vitro. However, there was no difference between the cells from WT mice and NOS2 - / - mice in response to IR exposure in vitro. Exposure of WT mice and NOS2 - / - mice to 6 Gy TBI decreased the white blood cell, red blood cell, and platelet counts in the peripheral blood and bone marrow mononuclear cells, and reduced the colony-forming ability of HPCs (P < 0.05), damaged the clonogenic function of HSCs. However, these changes were not significantly different in WT and NOS2 - / - mice. These data suggest that IR induces BM suppression in a NOS2-independent manner.

p27(kip1) Knockout enhances collateralization in response to hindlimb ischemia.

PubMed

Ankri-Eliahoo, Galit; Weitz, Kevin; Cox, Timothy C; Tang, Gale L

2016-05-01

The natural response to arterial occlusive disease is enlargement of collaterals; however, the molecular factors that control collateralization are not well understood. The gene p27(Kip1) (p27) affects human response to arterial injury. Previous studies have shown that overexpression of p27 inhibits vascular endothelial and vascular smooth muscle cell (VSMC) proliferation and angiogenesis. To test the hypothesis that knockout of p27 would improve collateralization in reaction to ischemia, we performed in vivo and in vitro experiments using p27 knockout (p27(-/-)) and wild-type (wt) mice. Hindlimb ischemia was induced by left femoral artery ligation in p27(-/-) and wt (C57BL/6) female mice. The mice underwent weekly laser Doppler perfusion imaging of the footpads until sacrifice on postoperative day 28 followed by microcomputed tomography scanning of both hindlimbs. VSMCs were isolated from p27(-/-) and wt mice and used in migration and gel contraction assays in the absence and presence of the nonspecific matrix metalloproteinase (MMP) inhibitor BB94. MMP-2 and MMP-9 messenger RNA (mRNA) expression was measured by quantitative reverse transcription-polymerase chain reaction in p27(-/-) and wt VSMCs. p27(-/-) mice reperfused more effectively than wt mice by laser Doppler starting from day 7 (ischemic/nonischemic ratio, 0.33 ± 0.02 vs 0.25 ± 0.02; P < .05) and continuing through day 28 (0.45 ± 0.04 vs 0.31 ± 0.04; P < .05). The gracilis collateral diameter was similar for the nonischemic hindlimbs of the p27(-/-) and wt mice, and this collateral pathway increased similarly after ischemia as assessed by microcomputed tomography. However, the p27(-/-) mice significantly enlarged a novel collateral pathway that bridged directly between the femoral artery proximal to the ligation site and the saphenous or popliteal artery distal to the ligation site more than wt mice (158 ± 18.3 vs 82 ± 22 μm; P < .001). p27(-/-) VSMCs migrated more (79% ± 5% vs 56% ± 6%; P < .05) and caused more gel contraction (18% ± 5% of the initial area vs 43% ± 4%; P < .05) than wt cells. Migration and collagen contraction were abolished in p27(-/-) and wt cells by MMP inhibition. p27(-/-) cells expressed significantly more MMP-2 mRNA than wt cells did. Knockout of p27 enhances arterial collateralization in response to hindlimb ischemia through enlargement of a new collateral pathway. In vitro, knockout of p27 increases collagen gel contraction in addition to stimulating VSMC migration. We speculate that p27 may affect collateralization through its role in regulating MMP-2 expression. Published by Elsevier Inc.

Study on the combined sewage sludge pyrolysis and gasification process: mass and energy balance.

PubMed

Wang, Zhonghui; Chen, Dezhen; Song, Xueding; Zhao, Lei

2012-12-01

A combined pyrolysis and gasification process for sewage sludge was studied in this paper for the purpose of its safe disposal with energy self-balance. Three sewage sludge samples with different dry basis lower heat values (LHV(db)) were used to evaluate the constraints on this combined process. Those samples were pre-dried and then pyrolysed within the temperature range of 400-550 degrees C. Afterwards, the char obtained from pyrolysis was gasified to produce fuel gas. The experimental results showed that the char yield ranged between 37.28 and 53.75 wt% of the dry sludge and it changed with ash content, pyrolysis temperature and LHV(db) of the sewage sludge. The gas from char gasification had a LHV around 5.31-5.65 MJ/Nm3, suggesting it can be utilized to supply energy in the sewage sludge drying and pyrolysis process. It was also found that energy balance in the combined process was affected by the LHV(db) of sewage sludge, moisture content and pyrolysis temperature. Higher LHV(db), lower moisture content and higher pyrolysis temperature benefit energy self-balance. For sewage sludge with a moisture content of 80 wt%, LHV(db) of sewage sludge should be higher than 18 MJ/kg and the pyrolysis temperature should be higher than 450 degrees C to maintain energy self-sufficiency when volatile from the pyrolysis process is the only energy supplier; when the LHV(db) was in the range of 14.65-18 MJ/kg, energy self-balance could be maintained in this combined process with fuel gas from char gasification as a supplementary fuel; auxiliary fuel was always needed if the LHV(db) was lower than 14.65 MJ/kg.

Generation of TCR-Expressing Innate Lymphoid-like Helper Cells that Induce Cytotoxic T Cell-Mediated Anti-leukemic Cell Response.

PubMed

Ueda, Norihiro; Uemura, Yasushi; Zhang, Rong; Kitayama, Shuichi; Iriguchi, Shoichi; Kawai, Yohei; Yasui, Yutaka; Tatsumi, Minako; Ueda, Tatsuki; Liu, Tian-Yi; Mizoro, Yasutaka; Okada, Chihiro; Watanabe, Akira; Nakanishi, Mahito; Senju, Satoru; Nishimura, Yasuharu; Kuzushima, Kiyotaka; Kiyoi, Hitoshi; Naoe, Tomoki; Kaneko, Shin

2018-06-05

CD4 + T helper (Th) cell activation is essential for inducing cytotoxic T lymphocyte (CTL) responses against malignancy. We reprogrammed a Th clone specific for chronic myelogenous leukemia (CML)-derived b3a2 peptide to pluripotency and re-differentiated the cells into original TCR-expressing T-lineage cells (iPS-T cells) with gene expression patterns resembling those of group 1 innate lymphoid cells. CD4 gene transduction into iPS-T cells enhanced b3a2 peptide-specific responses via b3a2 peptide-specific TCR. iPS-T cells upregulated CD40 ligand (CD40L) expression in response to interleukin-2 and interleukin-15. In the presence of Wilms tumor 1 (WT1) peptide, antigen-specific dendritic cells (DCs) conditioned by CD4-modified CD40L high iPS-T cells stimulated WT1-specific CTL priming, which eliminated WT1 peptide-expressing CML cells in vitro and in vivo. Thus, CD4 modification of CD40L high iPS-T cells generates innate lymphoid helper-like cells inducing bcr-abl-specific TCR signaling that mediates effectiveanti-leukemic CTL responses via DC maturation, showing potential for adjuvant immunotherapy against leukemia. Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

Inducible nitric oxide synthase during the late phase of sepsis is associated with hypothermia and immune cell migration.

PubMed

Takatani, Yudai; Ono, Kenji; Suzuki, Hiromi; Inaba, Masato; Sawada, Makoto; Matsuda, Naoyuki

2018-02-14

Hypothermia is a significant sign of sepsis, which is associated with poor prognosis, but few mechanisms underlying the regulation of hypothermia are known. Inducible nitric oxide synthase (iNOS) is a key inflammatory mediator of sepsis. However, the therapeutic benefit of iNOS inhibition in sepsis is still controversial, and requires elucidation in an accurate model system. In this study, wild-type (WT) mice showed temperature drops in a biphasic manner at the early and late phase of sepsis, and all mice died within 48 h of sepsis. In contrast, iNOS-knockout (KO) mice never showed the second temperature drop and exhibited improved mortality. Plasma nitric oxide (NO) levels of WT mice increased in the late phase of sepsis and correlated to hypothermia. The results indicate that iNOS-derived NO during the late phase of sepsis caused vasodilation-induced hypothermia and a lethal hypodynamic state. The expression of the iNOS mRNA was high in the lung of WT mice with sepsis, which reflects the pathology of acute respiratory distress syndrome (ARDS). We obtained the results in a modified keyhole-type cecal ligation and puncture model of septic shock induced by minimally invasive surgery. In this accurate and reproducible model system, we transplanted the bone marrow cells of GFP transgenic mice into WT and iNOS-KO mice, and evaluated the role of increased pulmonary iNOS expression in cell migration during the late phase of sepsis. We also investigated the quantity and type of bone marrow-derived cells (BMDCs) in the lung. The number of BMDCs in the lung of iNOS-KO mice was less than that in the lung of WT mice. The major BMDCs populations were CD11b-positive, iNOS-negative cells in WT mice, and Gr-1-positive cells in iNOS-KO mice that expressed iNOS. These results suggest that sustained hypothermia may be a beneficial guide for future iNOS-targeted therapy of sepsis, and that iNOS modulated the migratory efficiency and cell type of BMDCs in septic ARDS.

Paracellular transport through healthy and cystic fibrosis bronchial epithelial cell lines--do we have a proper model?

PubMed

Molenda, Natalia; Urbanova, Katarina; Weiser, Nelly; Kusche-Vihrog, Kristina; Günzel, Dorothee; Schillers, Hermann

2014-01-01

It has been reported recently that the cystic fibrosis transmembrane conductance regulator (CFTR) besides transcellular chloride transport, also controls the paracellular permeability of bronchial epithelium. The aim of this study was to test whether overexpressing wtCFTR solely regulates paracellular permeability of cell monolayers. To answer this question we used a CFBE41o- cell line transfected with wtCFTR or mutant F508del-CFTR and compered them with parental line and healthy 16HBE14o- cells. Transepithelial electrical resistance (TER) and paracellular fluorescein flux were measured under control and CFTR-stimulating conditions. CFTR stimulation significant decreased TER in 16HBE14o- and also in CFBE41o- cells transfected with wtCFTR. In contrast, TER increased upon stimulation in CFBE41o- cells and CFBE41o- cells transfected with F508del-CFTR. Under non-stimulated conditions, all four cell lines had similar paracellular fluorescein flux. Stimulation increased only the paracellular permeability of the 16HBE14o- cell monolayers. We observed that 16HBE14o- cells were significantly smaller and showed a different structure of cell-cell contacts than CFBE41o- and its overexpressing clones. Consequently, 16HBE14o- cells have about 80% more cell-cell contacts through which electrical current and solutes can leak. Also tight junction protein composition is different in 'healthy' 16HBE14o- cells compared to 'cystic fibrosis' CFBE41o- cells. We found that claudin-3 expression was considerably stronger in 16HBE14o- cells than in the three CFBE41o- cell clones and thus independent of the presence of functional CFTR. Together, CFBE41o- cell line transfection with wtCFTR modifies transcellular conductance, but not the paracellular permeability. We conclude that CFTR overexpression is not sufficient to fully reconstitute transport in CF bronchial epithelium. Hence, it is not recommended to use those cell lines to study CFTR-dependent epithelial transport.

Role of STAT1 in Chlamydia-Induced Type-1 Interferon Production in Oviduct Epithelial Cells

PubMed Central

Hosey, Kristen Lynette; Hu, Sishun

2015-01-01

We previously reported that Chlamydia muridarum-infected murine oviduct epithelial cells (OE cells) secrete interferon β (IFN-β) in a mostly TLR3-dependent manner. However, C. muridarum-infected TLR3-deficient OE cells were still able to secrete detectable levels of IFN-β into the supernatants, suggesting that other signaling pathways contribute to Chlamydia-induced IFN-β synthesis in these cells. We investigated the role of STAT1 as a possible contributor in the Chlamydia-induced type-1 IFN production in wild-type (WT) and TLR3-deficient OE cells to ascertain its putative role at early- and late-times during Chlamydia infection. Our data show that C. muridarum infection significantly increased STAT1 gene expression and protein activation in WT OE cells; however, TLR3-deficient OE cells showed diminished STAT1 protein activation and gene expression. There was significantly less IFN-β detected in the supernatants of C. muridarum-infected OE cells derived from mice deficient in STAT1 when compared with WT OE cells, which suggest that STAT1 is required for the optimal synthesis of IFN-β during infection. Real-time quantitative polymerase chain reaction analyses of signaling components of the type-1 IFN signaling pathway demonstrated equal upregulation in the expression of STAT2 and IRF7 genes in the WT and TLR3-deficient OE cells, but no upregulation in these genes in the STAT1-deficient OE cells. Finally, experiments in which INFAR1 was blocked with neutralizing antibody revealed that IFNAR1-mediated signaling was critical to the Chlamydia-induced upregulation in IFN-α gene transcription, but had no role in the Chlamydia-induced upregulation in IFN-β gene transcription. PMID:26262558

Tissue Non-specific Alkaline Phosphatase Expression is Needed for the Full Stimulation of T Cells and T Cell-Dependent Colitis.

PubMed

Hernández-Chirlaque, Cristina; Gámez-Belmonte, Reyes; Ocón, Borja; Martínez-Moya, Patricia; Wirtz, Stefan; Sánchez de Medina, Fermín; Martínez-Augustin, Olga

2017-07-01

Two alkaline phosphatase isoforms, intestinal [IAP] and tissue non-specific alkaline phosphatase [TNAP], are coexpressed in mouse colon, with the latter predominating in colitis. We aimed to examine the role of TNAP in T lymphocytes, using heterozygous TNAP+/- mice [as TNAP-/- mice are non-viable]. In vitro primary cultures and in vivo T cell models using TNAP+/- mice were used. Stimulated splenocytes [lipopolysaccharide and concanavalin A] and T lymphocytes [concanavalin A and a-CD3/a-CD28] showed a decreased cytokine production and expression when compared with wild-type [WT] cells. Decreased T cell activation was reproduced by the TNAP inhibitors levamisole, theophylline, and phenylalanine in WT cells. Intraperitoneal administration of anti-CD3 in vivo resulted in reduced plasma cytokine levels, and decreased activation of splenocytes and T cells ex vivo in TNAP+/- mice. We further tested the hypothesis that TNAP expressed in T lymphocytes is involved in T cell activation and inflammation, using the lymphocyte transfer model of colitis. Rag1-/- mice were transferred with T naïve cells [CD4+ CD62L+] from TNAP+/- or WT mice and developed colitis, which was attenuated in the group receiving TNAP+/- cells. Compared with WT, T cells from TNAP+/- mice showed a decreased capacity for proliferation, with no change in differentiation. Our results offer clear evidence that TNAP modulates T lymphocyte function and specifically T cell-dependent colitis. This was associated with distinct changes in the type of TNAP expressed, probably because of changes in glycosylation. Copyright © 2016 European Crohn’s and Colitis Organisation (ECCO). Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oup.com

γδT cells but not αβT cells contribute to sepsis-induced white matter injury and motor abnormalities in mice.

PubMed

Zhang, Xiaoli; Rocha-Ferreira, Eridan; Li, Tao; Vontell, Regina; Jabin, Darakhshan; Hua, Sha; Zhou, Kai; Nazmi, Arshed; Albertsson, Anna-Maj; Sobotka, Kristina; Ek, Joakim; Thornton, Claire; Hagberg, Henrik; Mallard, Carina; Leavenworth, Jianmei W; Zhu, Changlian; Wang, Xiaoyang

2017-12-20

Infection and sepsis are associated with brain white matter injury in preterm infants and the subsequent development of cerebral palsy. In the present study, we used a neonatal mouse sepsis-induced white matter injury model to determine the contribution of different T cell subsets (αβT cells and γδT cells) to white matter injury and consequent behavioral changes. C57BL/6J wild-type (WT), T cell receptor (TCR) δ-deficient (Tcrd -/- , lacking γδT cells), and TCRα-deficient (Tcra -/- , lacking αβT cells) mice were administered with lipopolysaccharide (LPS) at postnatal day (PND) 2. Brain myelination was examined at PNDs 12, 26, and 60. Motor function and anxiety-like behavior were evaluated at PND 26 or 30 using DigiGait analysis and an elevated plus maze. White matter development was normal in Tcrd -/- and Tcrα -/- compared to WT mice. LPS exposure induced reductions in white matter tissue volume in WT and Tcrα -/- mice, but not in the Tcrd -/- mice, compared with the saline-treated groups. Neither LPS administration nor the T cell deficiency affected anxiety behavior in these mice as determined with the elevated plus maze. DigiGait analysis revealed motor function deficiency after LPS-induced sepsis in both WT and Tcrα -/- mice, but no such effect was observed in Tcrd -/- mice. Our results suggest that γδT cells but not αβT cells contribute to sepsis-induced white matter injury and subsequent motor function abnormalities in early life. Modulating the activity of γδT cells in the early stages of preterm white matter injury might represent a novel therapeutic strategy for the treatment of perinatal brain injury.

Structures of native and affinity-enhanced WT1 epitopes bound to HLA-A*0201: implications for WT1-based cancer therapeutics.

PubMed

Borbulevych, Oleg Y; Do, Priscilla; Baker, Brian M

2010-09-01

Presentation of peptides by class I or class II major histocompatibility complex (MHC) molecules is required for the initiation and propagation of a T cell-mediated immune response. Peptides from the Wilms Tumor 1 transcription factor (WT1), upregulated in many hematopoetic and solid tumors, can be recognized by T cells and numerous efforts are underway to engineer WT1-based cancer vaccines. Here we determined the structures of the class I MHC molecule HLA-A*0201 bound to the native 126-134 epitope of the WT1 peptide and a recently described variant (R1Y) with improved MHC binding. The R1Y variant, a potential vaccine candidate, alters the positions of MHC charged side chains near the peptide N-terminus and significantly reduces the peptide/MHC electrostatic surface potential. These alterations indicate that the R1Y variant is an imperfect mimic of the native WT1 peptide, and suggest caution in its use as a therapeutic vaccine. Stability measurements revealed how the R1Y substitution enhances MHC binding affinity, and together with the structures suggest a strategy for engineering WT1 variants with improved MHC binding that retain the structural features of the native peptide/MHC complex. Copyright 2010 Elsevier Ltd. All rights reserved.

Piper betle induces phase I & II genes through Nrf2/ARE signaling pathway in mouse embryonic fibroblasts derived from wild type and Nrf2 knockout cells.

PubMed

Wan Hasan, Wan Nuraini; Kwak, Mi-Kyoung; Makpol, Suzana; Wan Ngah, Wan Zurinah; Mohd Yusof, Yasmin Anum

2014-02-23

Nuclear factor-erythroid 2 p45 related factor 2 (Nrf2) is a primary transcription factor, protecting cells from oxidative stress by regulating a number of antioxidants and phase II detoxifying enzymes. Dietary components such as sulforaphane in broccoli and quercetin in onions have been shown to be inducers of Nrf2. Piper betle (PB) grows well in tropical climate and the leaves are used in a number of traditional remedies for the treatment of stomach ailments and infections among Asians. The aim of this study was to elucidate the effect of Piper betle (PB) leaves extract in Nrf2 signaling pathway by using 2 types of cells; mouse embryonic fibroblasts (MEFs) derived from wild-type (WT) and Nrf2 knockout (N0) mice. WT and N0 cells were treated with 5 and 10 μg/ml of PB for 10 and 12-h for the determination of nuclear translocation of Nrf2 protein. Luciferase reporter gene activity was performed to evaluate the antioxidant response element (ARE)-induction by PB. Real-time PCR and Western blot were conducted on both WT and N0 cells after PB treatment for the determination of antioxidant enzymes [superoxide dismutase (SOD1) and heme-oxygenase (HO-1)], phase I oxidoreductase enzymes [ quinone oxidoreductase (NQO1)] and phase II detoxifying enzyme [glutathione S-transferase (GST)]. Nuclear translocation of Nrf2 by PB in WT cells was better after 10 h incubation compared to 12 h. Real time PCR and Western blot analysis showed increased expressions of Nrf2, NQO1 and GSTA1 genes with corresponding increases in glutathione, NQO1 and HO-1 proteins in WT cells. Reporter gene ARE was stimulated by PB as shown by ARE/luciferase assay. Interestingly, PB induced SOD1 gene and protein expressions in N0 cells but not in WT cells. The results of this study confirmed that PB activated Nrf2-ARE signaling pathway which subsequently induced some phase I oxidoreductase, phase II detoxifying and antioxidant genes expression via ARE reporter gene involved in the Nrf2 pathway with the exception of SOD1 which may not be dependent on this pathway.

Piper betle induces phase I & II genes through Nrf2/ARE signaling pathway in mouse embryonic fibroblasts derived from wild type and Nrf2 knockout cells

PubMed Central

2014-01-01

Background Nuclear factor-erythroid 2 p45 related factor 2 (Nrf2) is a primary transcription factor, protecting cells from oxidative stress by regulating a number of antioxidants and phase II detoxifying enzymes. Dietary components such as sulforaphane in broccoli and quercetin in onions have been shown to be inducers of Nrf2. Piper betle (PB) grows well in tropical climate and the leaves are used in a number of traditional remedies for the treatment of stomach ailments and infections among Asians. The aim of this study was to elucidate the effect of Piper betle (PB) leaves extract in Nrf2 signaling pathway by using 2 types of cells; mouse embryonic fibroblasts (MEFs) derived from wild-type (WT) and Nrf2 knockout (N0) mice. Methods WT and N0 cells were treated with 5 and 10 μg/ml of PB for 10 and 12-h for the determination of nuclear translocation of Nrf2 protein. Luciferase reporter gene activity was performed to evaluate the antioxidant response element (ARE)-induction by PB. Real-time PCR and Western blot were conducted on both WT and N0 cells after PB treatment for the determination of antioxidant enzymes [superoxide dismutase (SOD1) and heme-oxygenase (HO-1)], phase I oxidoreductase enzymes [NAD(P)H: quinone oxidoreductase (NQO1)] and phase II detoxifying enzyme [glutathione S-transferase (GST)]. Results Nuclear translocation of Nrf2 by PB in WT cells was better after 10 h incubation compared to 12 h. Real time PCR and Western blot analysis showed increased expressions of Nrf2, NQO1 and GSTA1 genes with corresponding increases in glutathione, NQO1 and HO-1 proteins in WT cells. Reporter gene ARE was stimulated by PB as shown by ARE/luciferase assay. Interestingly, PB induced SOD1 gene and protein expressions in N0 cells but not in WT cells. Conclusion The results of this study confirmed that PB activated Nrf2-ARE signaling pathway which subsequently induced some phase I oxidoreductase, phase II detoxifying and antioxidant genes expression via ARE reporter gene involved in the Nrf2 pathway with the exception of SOD1 which may not be dependent on this pathway. PMID:24559113

WT1 controls antagonistic FGF and BMP-pSMAD pathways in early renal progenitors.

PubMed

Motamedi, Fariba Jian; Badro, Danielle A; Clarkson, Michael; Lecca, M Rita; Bradford, Stephen T; Buske, Fabian A; Saar, Kathrin; Hübner, Norbert; Brändli, André W; Schedl, Andreas

2014-07-17

Kidney organogenesis requires the tight control of proliferation, differentiation and apoptosis of renal progenitor cells. How the balance between these cellular decisions is achieved remains elusive. The Wilms' tumour suppressor Wt1 is required for progenitor survival, but the molecular cause for renal agenesis in mutants is poorly understood. Here we demonstrate that lack of Wt1 abolishes fibroblast growth factor (FGF) and induces BMP/pSMAD signalling within the metanephric mesenchyme. Addition of recombinant FGFs or inhibition of pSMAD signalling rescues progenitor cell apoptosis induced by the loss of Wt1. We further show that recombinant BMP4, but not BMP7, induces an apoptotic response within the early kidney that can be suppressed by simultaneous addition of FGFs. These data reveal a hitherto unknown sensitivity of early renal progenitors to pSMAD signalling, establishes FGF and pSMAD signalling as antagonistic forces in early kidney development and places WT1 as a key regulator of pro-survival FGF signalling pathway genes.

Transient development of ovotestes in XX Sox9 transgenic mice.

PubMed

Gregoire, Elodie P; Lavery, Rowena; Chassot, Anne-Amandine; Akiyama, Haruhiko; Treier, Mathias; Behringer, Richard R; Chaboissier, Marie-Christine

2011-01-01

The sex of an individual results from the paternal transmission of the SRY gene located on the Y chromosome. In turn, SRY initiates Sox9 expression, a transcription factor required for testicular differentiation. Ectopic activation of SOX9 in XX Wt1:Sox9 transgenic mice induces female-to-male sex reversal in adult mice. Here we show that complete sex reversal is preceded by a transient phase of ovotestis differentiation with XX Wt1:Sox9 transgenic gonads containing a testicular central region and one or both ovarian poles indicating that Wt1:Sox9 is not as efficient as Sry to induce male development. In XX Wt1:Sox9(Tg/+) gonads, transgenic Sox9 is expressed earlier than Sox9 in XY gonads and is able to induce the expression of EGFP, knocked into the 3' UTR of Sox9 indicating that SOX9 is involved in the initiation and maintenance of its own expression. However, the delayed onset of expression of endogenous Sox9-EGFP suggests that this activation requires other factors, whose expression depends on SOX9. In the testicular regions of the XX Wt1:Sox9 ovotestes, proliferation of the XX fetal germ cells is hampered and they differentiate as pro-spermatogonia. This indicates that XX germ cells are not competent to respond to proliferative signals released from a testicular environment. In the ovarian regions, despite the continuous mRNA expression of the WT1:Sox9 transgene, the SOX9 protein does not accumulate suggesting that regulation of this gene in ovarian cells involves post-transcriptional mechanisms. Finally, ovarian cells of the XX Wt1:Sox9 ovotestis undergo apoptosis during late embryogenesis leading to complete female-to-male sex reversal of the transgenic mice at birth. Copyright © 2010 Elsevier Inc. All rights reserved.

Nutlin‐3a selects for cells harbouring TP 53 mutations

PubMed Central

Hollstein, Monica; Arlt, Volker M.; Phillips, David H.

2016-01-01

TP53 mutations occur in half of all human tumours. Mutagen‐induced or spontaneous TP53 mutagenesis can be studied in vitro using the human TP53 knock‐in (Hupki) mouse embryo fibroblast (HUF) immortalisation assay (HIMA). TP53 mutations arise in up to 30% of mutagen‐treated, immortalised HUFs; however, mutants are not identified until TP53 sequence analysis following immortalisation (2–5 months) and much effort is expended maintaining TP53‐WT cultures. In order to improve the selectivity of the HIMA for HUFs harbouring TP53 mutations, we explored the use of Nutlin‐3a, an MDM2 inhibitor that leads to stabilisation and activation of wild‐type (WT) p53. First, we treated previously established immortal HUF lines carrying WT or mutated TP53 with Nutlin‐3a to examine the effect on cell growth and p53 activation. Nutlin‐3a induced the p53 pathway in TP53‐WT HUFs and inhibited cell growth, whereas most TP53‐mutated HUFs were resistant to Nutlin‐3a. We then assessed whether Nutlin‐3a treatment could discriminate between TP53‐WT and TP53‐mutated cells during the HIMA (n = 72 cultures). As immortal clones emerged from senescent cultures, each was treated with 10 µM Nutlin‐3a for 5 days and observed for sensitivity or resistance. TP53 was subsequently sequenced from all immortalised clones. We found that all Nutlin‐3a‐resistant clones harboured TP53 mutations, which were diverse in position and functional impact, while all but one of the Nutlin‐3a‐sensitive clones were TP53‐WT. These data suggest that including a Nutlin‐3a counter‐screen significantly improves the specificity and efficiency of the HIMA, whereby TP53‐mutated clones are selected prior to sequencing and TP53‐WT clones can be discarded. PMID:27813088

Nitrification in histosols: a potential role for the heterotrophic nitrifier.

PubMed

Tate, R L

1977-04-01

Insufficient populations of Nitrosomonas and Nitrobacter were found in a Pahokee muck soil (Lithic medidaprit) to account for the nitrate concentration observed. To determine if heterotrophic nitrifiers could account for some of this discrepancy, a method was developed to measure the levels of heterotrophic nitrifiers in soil. A population of 4.1 X 10(5) Arthrobacter per g of dry fallow soil, capable of producing nitrite and/or nitrate from reduced nitrogenous compounds, was observed. Amendment of the much with 0.5% (wt/wt) sodium acetate and 0.1% (wt/wt) ammonium-nitrogen as ammonium sulfate (final concentrations) not only resulted in the usual increase in autotrophic nitrifiers, but also in a fourfold increase in the heterotrophic nitrifying Arrthrobacter. Amendment of like samples with N-Serve [2-chloro-6(trichloromethyl) pyridinel] prevented the increase in Nitrosomonas, but not that in the heterotrophic nitrifiers. Nitrate production in the presence of the inhibitor was diminished but not prevented. An Arthrobacter sp., isolated from the muck, produced nitrite when inoculated at high densities into sterile soil, unamended or amended with sodium acetate and/or ammomium sulfate. These data suggest that the heterotrophic population may be responsible for some of the nitrate produced in these Histosols.

Myeloid-derived suppressor cells regulate T cell and B cell responses during autoimmune disease.

PubMed

Crook, Kristen R; Jin, Mengyao; Weeks, Michael F; Rampersad, Rishi R; Baldi, Robert M; Glekas, Amy S; Shen, Yajuan; Esserman, Denise A; Little, Paul; Schwartz, Todd A; Liu, Peng

2015-03-01

MDSCs are a heterogeneous group of myeloid cells that suppress T cell activity in cancer and autoimmune disease. The effect of MDSCs on B cell function is not clear. Using the CIA model of autoimmune disease, we found an increase in M-MDSCs in the periphery of WT mice with CIA compared with naïve mice. These MDSCs were absent from the periphery of CCR2(-/-) mice that developed exacerbated disease. M-MDSCs, isolated from immunized mice, inhibited autologous CD4(+) T cell proliferation. The M-MDSC-mediated suppression of T cell proliferation was NO and IFN-γ dependent but IL-17 independent. Furthermore, we demonstrated for the first time that M-MDSCs from CIA mice also inhibited autologous B cell proliferation and antibody production. The suppression of B cells by M-MDSCs was dependent on the production of NO and PGE2 and required cell-cell contact. Administration of M-MDSCs rescued CCR2(-/-) mice from the exacerbated CIA phenotype and ameliorated disease in WT mice. Furthermore, adoptive transfer of M-MDSCs reduced autoantibody production by CCR2(-/-) and WT mice. In summary, M-MDSCs inhibit T cell and B cell function in CIA and may serve as a therapeutic approach in the treatment of autoimmune arthritis. © Society for Leukocyte Biology.

Comprehensive process maps for synthesizing high density aluminum oxide-carbon nanotube coatings by plasma spraying for improved mechanical and wear properties

NASA Astrophysics Data System (ADS)

Keshri, Anup Kumar

Plasma sprayed aluminum oxide ceramic coating is widely used due to its outstanding wear, corrosion, and thermal shock resistance. But porosity is the integral feature in the plasma sprayed coating which exponentially degrades its properties. In this study, process maps were developed to obtain Al2O3-CNT composite coatings with the highest density (i.e. lowest porosity) and improved mechanical and wear properties. Process map is defined as a set of relationships that correlates large number of plasma processing parameters to the coating properties. Carbon nanotubes (CNTs) were added as reinforcement to Al2O 3 coating to improve the fracture toughness and wear resistance. Two novel powder processing approaches viz spray drying and chemical vapor growth were adopted to disperse CNTs in Al2O3 powder. The degree of CNT dispersion via chemical vapor deposition (CVD) was superior to spray drying but CVD could not synthesize powder in large amount. Hence optimization of plasma processing parameters and process map development was limited to spray dried Al2O3 powder containing 0, 4 and 8 wt. % CNTs. An empirical model using Pareto diagram was developed to link plasma processing parameters with the porosity of coating. Splat morphology as a function of plasma processing parameter was also studied to understand its effect on mechanical properties. Addition of a mere 1.5 wt. % CNTs via CVD technique showed ˜27% and ˜24% increase in the elastic modulus and fracture toughness respectively. Improved toughness was attributed to combined effect of lower porosity and uniform dispersion of CNTs which promoted the toughening by CNT bridging, crack deflection and strong CNT/Al2O3 interface. Al2O 3-8 wt. % CNT coating synthesized using spray dried powder showed 73% improvement in the fracture toughness when porosity reduced from 4.7% to 3.0%. Wear resistance of all coatings at room and elevated temperatures (573 K, 873 K) showed improvement with CNT addition and decreased porosity. Such behavior was due to improved mechanical properties, protective film formation due to tribochemical reaction, and CNT bridging between the splats. Finally, process maps correlating porosity content, CNT content, mechanical properties, and wear properties were developed.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Pickenheim, B.; Hansen, E.; Leishear, R.

A 10-inch READCO mixer is used for mixing the premix (45 (wt%) fly ash, 45 wt% slag, and 10 wt% portland cement) with salt solution in the Saltstone Production Facility (SPF). The Saltstone grout free falls into the grout hopper which feeds the suction line leading to the Watson SPX 100 duplex hose pump. The Watson SPX 100 pumps the grout through approximately 1500 feet of piping prior to being discharged into the Saltstone Disposal Facility (SDF) vaults. The existing grout hopper has been identified by the Saltstone Enhanced Low Activity Waste Disposal (ELAWD) project for re-design. The current nominalmore » working volume of this hopper is 12 gallons and does not permit handling an inadvertent addition of excess dry feeds. Saltstone Engineering has proposed a new hopper tank that will have a nominal working volume of 300 gallons and is agitated with a mechanical agitator. The larger volume hopper is designed to handle variability in the output of the READCO mixer and process upsets without entering set back during processing. The objectives of this task involve scaling the proposed hopper design and testing the scaled hopper for the following processing issues: (1) The effect of agitation on radar measurement. Formation of a vortex may affect the ability to accurately measure the tank level. The agitator was run at varying speeds and with varying grout viscosities to determine what parameters cause vortex formation and whether measurement accuracy is affected. (2) A dry feeds over addition. Engineering Calculating X-ESR-Z-00017 1 showed that an additional 300 pounds of dry premix added to a 300 gallon working volume would lower the water to premix ratio (W/P) from the nominal 0.60 to 0.53 based on a Salt Waste Processing Facility (SWPF) salt simulant. A grout with a W/P of 0.53 represents the upper bound of grout rheology that could be processed at the facility. A scaled amount of dry feeds will be added into the hopper to verify that this is a recoverable situation. (3) The necessity of baffles in the hopper. The preference of the facility is not to have baffles in the hopper; however, if the initial testing indicates inadequate agitation or difficulties with the radar measurement, baffles will be tested.« less

Coenzyme Q(1) as a probe for mitochondrial complex I activity in the intact perfused hyperoxia-exposed wild-type and Nqo1-null mouse lung.

PubMed

Bongard, Robert D; Myers, Charles R; Lindemer, Brian J; Baumgardt, Shelley; Gonzalez, Frank J; Merker, Marilyn P

2012-05-01

Previous studies showed that coenzyme Q(1) (CoQ(1)) reduction on passage through the rat pulmonary circulation was catalyzed by NAD(P)H:quinone oxidoreductase 1 (NQO1) and mitochondrial complex I, but that NQO1 genotype was not a factor in CoQ(1) reduction on passage through the mouse lung. The aim of the present study was to evaluate the complex I contribution to CoQ(1) reduction in the isolated perfused wild-type (NQO1(+/+)) and Nqo1-null (NQO1(-)/(-)) mouse lung. CoQ(1) reduction was measured as the steady-state pulmonary venous CoQ(1) hydroquinone (CoQ(1)H(2)) efflux rate during infusion of CoQ(1) into the pulmonary arterial inflow. CoQ(1)H(2) efflux rates during infusion of 50 μM CoQ(1) were not significantly different for NQO1(+/+) and NQO1(-/-) lungs (0.80 ± 0.03 and 0.68 ± 0.07 μmol·min(-1)·g lung dry wt(-1), respectively, P > 0.05). The mitochondrial complex I inhibitor rotenone depressed CoQ(1)H(2) efflux rates for both genotypes (0.19 ± 0.08 and 0.08 ± 0.04 μmol·min(-1)·g lung dry wt(-1) for NQO1(+/+) and NQO1(-/-), respectively, P < 0.05). Exposure of mice to 100% O(2) for 48 h also depressed CoQ(1)H(2) efflux rates in NQO1(+/+) and NQO1(-/-) lungs (0.43 ± 0.03 and 0.11 ± 0.04 μmol·min(-1)·g lung dry wt(-1), respectively, P < 0.05 by ANOVA). The impact of rotenone or hyperoxia on CoQ(1) redox metabolism could not be attributed to effects on lung wet-to-dry weight ratios, perfusion pressures, perfused surface areas, or total venous effluent CoQ(1) recoveries, the latter measured by spectrophotometry or mass spectrometry. Complex I activity in mitochondria-enriched lung fractions was depressed in hyperoxia-exposed lungs for both genotypes. This study provides new evidence for the potential utility of CoQ(1) as a nondestructive indicator of the impact of pharmacological or pathological exposures on complex I activity in the intact perfused mouse lung.

Nanostructured composite material graphite/TiO2 and its antibacterial activity under visible light irradiation.

PubMed

Dědková, Kateřina; Lang, Jaroslav; Matějová, Kateřina; Peikertová, Pavlína; Holešinský, Jan; Vodárek, Vlastimil; Kukutschová, Jana

2015-08-01

The paper addresses laboratory preparation, characterization and in vitro evaluation of antibacterial activity of graphite/TiO2 nanocomposites. Composites graphite/TiO2 with various ratio of TiO2 nanoparticles (30wt.%, and 50wt.%) to graphite were prepared using a thermal hydrolysis of titanylsulfate in the presence of graphite particles, and subsequently dried at 80°C. X-ray powder diffraction, transmission electron microscopy and Raman microspectroscopy served as phase-analytical methods distinguishing anatase and rutile phases in the prepared composites. Scanning and transmission electron microscopy techniques were used for characterization of morphology of the prepared samples. A developed modification of the standard microdilution test was used for in vitro evaluation of daylight induced antibacterial activity, using four common human pathogenic bacterial strains (Staphylococcus aureus, Escherichia coli, Enterococcus faecalis and Pseudomonas aeruginosa). Antibacterial activity of the graphite/TiO2 nanocomposites could be based mainly on photocatalytic reaction with subsequent potential interaction of reactive oxygen species with bacterial cells. During the antibacterial activity experiments, the graphite/TiO2 nanocomposites exhibited antibacterial activity, where differences in the onset of activity and activity against bacterial strains were observed. The highest antibacterial activity evaluated as minimum inhibitory concentration was observed against P. aeruginosa after 180min of irradiation. Copyright © 2015 Elsevier B.V. All rights reserved.

Mercury concentrations in Southern Beaufort Sea polar bears: variation based on stable isotopes of carbon and nitrogen.

PubMed

Cardona-Marek, Tamara; Knott, Katrina K; Meyer, Benjamin E; O'Hara, Todd M

2009-07-01

Total Hg concentration was measured in hair and whole blood of 52 adult Southern Beaufort Sea polar bears (Ursus maritimus) captured in the spring of 2005. Stable isotopic signatures (i.e., 13C/12C, delta13C; 15N/14N, delta15N) in hair and two blood compartments (packed blood cells/clot and serum) were determined to assess the variation of Hg concentrations among polar bears in relation to their feeding ecology and other biological factors. Concentrations of Hg in hair and blood (2.2-23.9 microg/g dry wt and 0.007-0.213 microg/g wet wt, respectively) were within the range of values previously reported for polar bears in Canada and East Greenland. Mercury concentration in hair from females was higher than that in hair from males, and concentration was related to interactions between delta13C, delta15N, and longitude of capture location. Mercury concentrations in hair were inversely correlated to delta13C in hair and blood, suggesting that polar bears with greater total Hg concentrations fed more on pelagic prey, such as ringed seals or beluga whale, than on benthic prey. Variability in Hg concentrations in polar bear hair and blood may be the result of intraspecific or regional variation in prey selection rather than strictly trophic level interactions.

Enhanced poly(3-hydroxypropionate) production via β-alanine pathway in recombinant Escherichia coli

PubMed Central

Lacmata, Stephen Tamekou; Kuiate, Jules-Roger; Ding, Yamei; Xian, Mo; Liu, Huizhou; Boudjeko, Thaddée; Feng, Xinjun; Zhao, Guang

2017-01-01

Poly(3-hydroxypropionate) (P3HP) is a thermoplastic with great compostability and biocompatibility, and can be produced through several biosynthetic pathways, in which the glycerol pathway achieved the highest P3HP production. However, exogenous supply of vitamin B12 was required to maintain the activity of glycerol dehydratase, resulting in high production cost. To avoid the addition of VB12, we have previously constructed a P3HP biosynthetic route with β-alanine as intermediate, and the present study aimed to improve the P3HP production of this pathway. L-aspartate decarboxylase PanD was found to be the rate-limiting enzyme in the β-alanine pathway firstly. To improve the pathway efficiency, PanD was screened from four different sources (Escherichia coli, Bacillus subtilis, Pseudomonas fluorescens, and Corynebacterium glutamicum). And PanD from C. glutamicum was found to have the highest activity, the P3HP production was improved in flask cultivation with this enzyme. To further improve the production, the host strain was screened and the culture condition was optimized. Under optimal conditions, production and content of P3HP reached to 10.2 g/L and 39.1% (wt/wt [cell dry weight]) in an aerobic fed-batch fermentation. To date, this is the highest P3HP production without VB12. PMID:28253372

Effects of Coating Materials and Processing Conditions on Flow Enhancement of Cohesive Acetaminophen Powders by High-Shear Processing With Pharmaceutical Lubricants.

PubMed

Wei, Guoguang; Mangal, Sharad; Denman, John; Gengenbach, Thomas; Lee Bonar, Kevin; Khan, Rubayat I; Qu, Li; Li, Tonglei; Zhou, Qi Tony

2017-10-01

This study has investigated the surface coating efficiency and powder flow improvement of a model cohesive acetaminophen powder by high-shear processing with pharmaceutical lubricants through 2 common equipment, conical comil and high-shear mixer. Effects of coating materials and processing parameters on powder flow and surface coating coverage were evaluated. Both Carr's index and shear cell data indicated that processing with the lubricants using comil or high-shear mixer substantially improved the flow of the cohesive acetaminophen powder. Flow improvement was most pronounced for those processed with 1% wt/wt magnesium stearate, from "cohesive" for the V-blended sample to "easy flowing" for the optimally coated sample. Qualitative and quantitative characterizations demonstrated a greater degree of surface coverage for high-shear mixing compared with comilling; nevertheless, flow properties of the samples at the corresponding optimized conditions were comparable between 2 techniques. Scanning electron microscopy images demonstrated different coating mechanisms with magnesium stearate or l-leucine (magnesium stearate forms a coating layer and leucine coating increases surface roughness). Furthermore, surface coating with hydrophobic magnesium stearate did not retard the dissolution kinetics of acetaminophen. Future studies are warranted to evaluate tableting behavior of such dry-coated pharmaceutical powders. Copyright © 2017 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.

Down-regulation of MutS homolog 3 by hypoxia in human colorectal cancer

PubMed Central

Li, Jie; Koike, Junichi; Kugoh, Hiroyuki; Arita, Michitsune; Ohhira, Takahito; Kikuchi, Yoshinori; Funahashi, Kimihiko; Takamatsu, Ken; Boland, C. Richard; Koi, Minoru; Hemmi, Hiromichi

2013-01-01

Down-regulation of hMSH3 is associated with elevated microsatellite alterations at selected tetranucleotide repeats and low levels of microsatellite instability in colorectal cancer (CRC). However, the mechanism that down-regulates hMSH3 in CRC is not known. In this study, a significant association between over-expression of glucose transporter 1, a marker for hypoxia, and down-regulation of hMSH3 in CRC tissues was observed. Therefore, we examined the effect of hypoxia on the expression of hMSH3 in human cell lines. When cells with wild type p53 (wt-p53) were exposed to hypoxia, rapid down-regulation of both hMSH2 and hMSH3 occurred. In contrast, when null or mutated p53 (null/mut-p53) cells were exposed to hypoxia, only hMSH3 was down-regulated, and at slower rate than wt-p53 cells. Using a reporter assay, we found that disruption of the two putative hypoxia response elements (HREs) located within the promoter region of the hMSH3 abrogated the suppressive effect of hypoxia on reporter activity regardless of p53 status. In an EMSA, two different forms of HIF-1α complexes that specifically bind to these HREs were detected. A larger complex containing HIF-1α predominantly bound to the HREs in hypoxic null/mut-p53 cells whereas a smaller complex predominated in wt-p53 cells. Finally, HIF-1α knockdown by siRNA significantly inhibited down-regulation of hMSH3 by hypoxia in both wt-p53 and mut-p53 cells. Taken together, our results suggest that the binding of HIF-1α complexes to HRE sites is necessary for down-regulation of hMSH3 in both wt-p53 and mut-p53 cells. PMID:22343000

Human Langerhans cells use an IL-15R-α/IL-15/pSTAT5-dependent mechanism to break T-cell tolerance against the self-differentiation tumor antigen WT1.

PubMed

Romano, Emanuela; Cotari, Jesse W; Barreira da Silva, Rosa; Betts, Brian C; Chung, David J; Avogadri, Francesca; Fink, Mitsu J; St Angelo, Erin T; Mehrara, Babak; Heller, Glenn; Münz, Christian; Altan-Bonnet, Gregoire; Young, James W

2012-05-31

Human CD34(+) progenitor-derived Langerhans-type dendritic cells (LCs) are more potent stimulators of T-cell immunity against tumor and viral antigens in vitro than are monocyte-derived DCs (moDCs). The exact mechanisms have remained elusive until now, however. LCs synthesize the highest amounts of IL-15R-α mRNA and protein, which binds IL-15 for presentation to responder lymphocytes, thereby signaling the phosphorylation of signal transducer and activator of transcription 5 (pSTAT5). LCs electroporated with Wilms tumor 1 (WT1) mRNA achieve sufficiently sustained presentation of antigenic peptides, which together with IL-15R-α/IL-15, break tolerance against WT1 by stimulating robust autologous, WT1-specific cytolytic T-lymphocytes (CTLs). These CTLs develop from healthy persons after only 7 days' stimulation without exogenous cytokines and lyse MHC-restricted tumor targets, which include primary WT1(+) leukemic blasts. In contrast, moDCs require exogenous rhuIL-15 to phosphorylate STAT5 and attain stimulatory capacity comparable to LCs. LCs therefore provide a more potent costimulatory cytokine milieu for T-cell activation than do moDCs, thus accounting for their superior stimulation of MHC-restricted Ag-specific CTLs without need for exogenous cytokines. These data support the use of mRNA-electroporated LCs, or moDCs supplemented with exogenous rhuIL-15, as vaccines for cancer immunotherapy to break tolerance against self-differentiation antigens shared by tumors.

Study of SEM preparation artefacts with correlative microscopy: Cell shrinkage of adherent cells by HMDS-drying.

PubMed

Katsen-Globa, Alisa; Puetz, Norbert; Gepp, Michael M; Neubauer, Julia C; Zimmermann, Heiko

2016-11-01

One of the often reported artefacts during cell preparation to scanning electron microscopy (SEM) is the shrinkage of cellular objects, that mostly occurs at a certain time-dependent stage of cell drying. Various methods of drying for SEM, such as critical point drying, freeze-drying, as well as hexamethyldisilazane (HMDS)-drying, were usually used. The latter becomes popular since it is a low cost and fast method. However, the correlation of drying duration and real shrinkage of objects was not investigated yet. In this paper, cell shrinkage at each stage of preparation for SEM was studied. We introduce a shrinkage coefficient using correlative light microscopy (LM) and SEM of the same human mesenchymal stem cells (hMSCs). The influence of HMDS-drying duration on the cell shrinkage is shown: the longer drying duration, the more shrinkage is observed. Furthermore, it was demonstrated that cell shrinkage is inversely proportional to cultivation time: the longer cultivation time, the more cell spreading area and the less cell shrinkage. Our results can be applicable for an exact SEM quantification of cell size and determination of cell spreading area in engineering of artificial cellular environments using biomaterials. SCANNING 38:625-633, 2016. © 2016 Wiley Periodicals, Inc. © Wiley Periodicals, Inc.

Differential regulation of insulin-like growth factor-I receptor gene expression by wild type and mutant androgen receptor in prostate cancer cells.

PubMed

Schayek, Hagit; Seti, Hila; Greenberg, Norman M; Sun, Shihua; Werner, Haim; Plymate, Stephen R

2010-07-29

The progression of prostate cancer from an organ-confined, androgen-sensitive disease to a metastatic one is associated with dysregulation of androgen receptor (AR)-regulated target genes and with a decrease in insulin-like growth factor-I receptor (IGF-IR) expression. To investigate the differential effects of wild type (wt) and mutant AR on IGF-IR levels we employed a series of isogenic prostate-derived cell lines and human xenografts. We show that basal and phosphorylated IGF-IR levels progressively decreased as prostate cancer cells became more tumorigenic and metastatic. In addition, we show that wt, but not mutant, AR along with dihydrotestosterone treatment increased IGF-IR promoter activity and endogenous IGF-IR levels. ChIP analysis show enhanced AR binding to the IGF-IR promoter in AR-overexpressing cells. Finally, wt AR-overexpressing cells display an enhanced proliferation rate. In summary, we provide evidence that activated wt AR enhances IGF-IR transcription in prostate cancer cells via a mechanism that involves AR binding to the IGF-IR promoter. AR mutations alter the ability of the mutated protein to regulate IGF-IR expression. Our results suggest that prostate cancer progression is associated with a decrease in IGF-IR expression that could be the result of impaired ability of AR to stimulate IGF-IR gene expression. 2010 Elsevier Ireland Ltd. All rights reserved.

Differential regulation of insulin-like growth factor-I receptor gene expression by wild type and mutant androgen receptor in prostate cancer cells

PubMed Central

Schayek, Hagit; Seti, Hila; Greenberg, Norman M.; Sun, Shihua; Werner, Haim; Plymate, Stephen R.

2010-01-01

The progression of prostate cancer from an organ-confined, androgen-sensitive disease to a metastatic one is associated with dysregulation of androgen receptor (AR)-regulated target genes and with a decrease in insulin-like growth factor-I receptor (IGF-IR) expression. To investigate the differential effects of wild type (wt) and mutant AR on IGF-IR levels we employed a series of isogenic prostate-derived cell lines and human xenografts. We show that basal and phosphorylated IGF-IR levels progressively decreased as prostate cancer cells became more tumorigenic and metastatic. In addition, we show that wt, but not mutant, AR along with dihydrotestosterone treatment increased IGF-IR promoter activity and endogenous IGF-IR levels. ChIP analysis show enhanced AR binding to the IGF-IR promoter in AR-overexpressing cells. Finally, wt AR-overexpressing cells display an enhanced proliferation rate. In summary, we provide evidence that activated wt AR enhances IGF-IR transcription in prostate cancer cells via a mechanism that involves AR binding to the IGF-IR promoter. AR mutations alter the ability of the mutated protein to regulate IGF-IR expression. Our results suggest that prostate cancer progression is associated with a decrease in IGF-IR expression that could be the result of impaired ability of AR to stimulate IGF-IR gene expression. PMID:20417685

Randomized Phase II Trial of Adjuvant WT-1 Analog Peptide Vaccine in Patients with Malignant Pleural Mesothelioma after Completion of Multimodality Therapy

DTIC Science & Technology

2017-11-01

journal of cancer research : Gann 1999; 90(2): 194-204. 6. Rosenfeld C, Cheever MA, Gaiger A. WT1 in acute leukemia, chronic myelogenous leukemia...and myelodysplastic syndrome : therapeutic potential of WT1 targeted therapies. Leukemia 2003; 17(7): 1301-12. 7. Cheever MA, Allison JP, Ferris AS...Vaccination with synthetic analog peptides derived from WT1 oncoprotein induces T-cell responses in patients with complete remission from acute myeloid

Electrochemical Characteristics of Cell Cultured Ti-Nb-Zr Alloys After Nano-Crystallized Si-HA Coating.

PubMed

Jeong, Yong-Hoon; Choe, Han-Cheol

2015-01-01

The aim of this study was to investigate the electrochemical characteristics of nano crystallized Si-HA coating on Ti-Nb-Zr alloy after human osteoblast like (HOB) cell attachment. The Ti-Nb-Zr alloy was manufactured with 35 wt.% of Nb and 10 wt.% of Zr by arc melting furnace to appropriate physical properties as biomaterials. The HA and Si-substituted coatings were prepared by electron-beam physical vapor deposition method with 0.5, 0.8 and 1.2 wt.% of Si contents, and nano aging treatment was performed 500 degrees C for 1 h. The characteristics of coating surface were analyzed by field emission scanning electron microscopy, energy dispersive X-ray spectroscopy, and X-ray diffraction, respectively. To evaluate of cell attachment on cell cultured surface, the potentiodynamic test was performed on the surface using HOB cells. The results showed that the Si-HA coating surface showed higher tendency of cell attachment than that of single HA coating, corrosion resistance value was increased by dense of cell attachment.

Inhibition of thrombin receptor signaling on α-smooth muscle actin(+) CD34(+) progenitors leads to repair after murine immune vascular injury.

PubMed

Chen, Daxin; Shrivastava, Seema; Ma, Liang; Tham, El-Li; Abrahams, Joel; Coe, J David; Scott, Diane; Lechler, Robert I; McVey, John H; Dorling, Anthony

2012-01-01

The goal of this study was to use mice expressing human tissue factor pathway inhibitor (TFPI) on α-smooth muscle actin (α-SMA)(+) cells as recipients of allogeneic aortas to gain insights into the cellular mechanisms of intimal hyperplasia (IH). BALB/c aortas (H-2(d)) transplanted into α-TFPI-transgenic (Tg) mice (H-2(b)) regenerated a quiescent endothelium in contrast to progressive IH seen in C57BL/6 wild-type (WT) mice even though both developed aggressive anti-H-2(d) alloresponses, indicating similar vascular injuries. Adoptively transferred Tg CD34(+) (but not CD34(-)) cells inhibited IH in WT recipients, indicating the phenotype of α-TFPI-Tg mice was due to these cells. Compared with syngeneic controls, endogenous CD34(+) cells were mobilized in significant numbers after allogeneic transplantation, the majority showing sustained expression of tissue factor and protease-activated receptor-1 (PAR-1). In WT, most were CD45(+) myeloid progenitors coexpressing CD31, vascular endothelial growth factor receptor-2 and E-selectin; 10% of these cells coexpressed α-SMA and were recruited to the neointima. In contrast, the α-SMA(+) human TFPI(+) CD34(+) cells recruited in Tg recipients were from a CD45(-) lineage. WT CD34(+) cells incubated with a PAR-1 antagonist or taken from PAR-1-deficient mice inhibited IH as Tg cells did. Specific inhibition of thrombin generation or PAR-1 signaling on α-SMA(+) CD34(+) cells inhibits IH and promotes regenerative repair despite ongoing immune-mediated damage.

Polycyclic aromatic and aliphatic hydrocarbons in Chukchi Sea biota and sediments and their toxicological response in the Arctic cod, Boreogadus saida

NASA Astrophysics Data System (ADS)

Harvey, H. Rodger; Taylor, Karen A.; Pie, Hannah V.; Mitchelmore, Carys L.

2014-04-01

As part of the Chukchi Sea Offshore Monitoring in Drilling Area-Chemical and Benthos (COMIDA CAB) project, we determined the distribution and concentrations of aliphatic n-alkanes and polycyclic aromatic hydrocarbons (PAHs) in surface sediments (0-1 cm) among 52 sites across the Chukchi Sea and in muscle tissues of the benthic Northern whelk, Neptunea heros, collected opportunistically. In addition, downcore profiles of contaminants were determined at three targeted sites to establish historic patterns. Baseline responses of PAH exposure and its potential toxicological effects were examined in the common Arctic cod, Boreogadus saida, through measures of cytochrome P4501A/ ethoxyresorufin O-deethylase (CYP1A/EROD), glutathione-S-transferase (GST), and Cu/Zn superoxide dismutase (SOD) activity in liver tissue. The total concentration of PAHs in surface sediments throughout the study area, including parent and alkyl-homologs, were very low (<1600 ng g-1 dry wt) except for a single station, where values were 2-20-fold greater than at other baseline sites (2956 ng g-1 dry wt). Alkyl-substituted PAHs were the dominant form in all surface (54-93%) and subsurface sediments (50-81% of the total), with a general decrease in total PAH concentrations observed downcore. In biota, larger Neptunea showed lower total concentrations of PAHs in foot muscles (4.5-10.7 ng g-1 wet wt) compared to smaller animals; yet aliphatic n-alkane (C19-C33) concentrations (0.655-5.20 μg g-1 wet wt) increased in larger organisms with distributions dominated by long-chain (C23-C33) hydrocarbons. In B. saida, CYP1A1, GST, and SOD enzyme levels were comparable to baseline levels previously reported in other pristine systems. Of the three assays, only SOD had a significant correlation between gene expression and enzyme activity.

Safety and tolerability of allogeneic dendritic cell vaccination with induction of Wilms tumor 1-specific T cells in a pediatric donor and pediatric patient with relapsed leukemia: a case report and review of the literature.

PubMed

Saito, Shoji; Yanagisawa, Ryu; Yoshikawa, Kentaro; Higuchi, Yumiko; Koya, Terutsugu; Yoshizawa, Kiyoshi; Tanaka, Miyuki; Sakashita, Kazuo; Kobayashi, Takashi; Kurata, Takashi; Hirabayashi, Koichi; Nakazawa, Yozo; Shiohara, Masaaki; Yonemitsu, Yoshikazu; Okamoto, Masato; Sugiyama, Haruo; Koike, Kenichi; Shimodaira, Shigetaka

2015-03-01

A 15-year-old girl with acute lymphoblastic leukemia received allogeneic dendritic cell vaccination, pulsed with Wilms tumor 1 (WT1) peptide, after her third hematopoietic stem cell transplantation (HSCT). The vaccines were generated from the third HSCT donor, who was her younger sister, age 12 years. The patient received 14 vaccines and had no graft-versus-host disease or systemic adverse effect, aside from grade 2 skin reaction at the injection site. WT1-specific immune responses were detected after vaccination by both WT1-tetramer analysis and enzyme-linked immunosorbent spot assay. This strategy may be safe, tolerable and even feasible for patients with a relapse after HSCT. Copyright © 2015 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

The yeast Holliday junction resolvase, CCE1, can restore wild-type mitochondrial DNA to human cells carrying rearranged mitochondrial DNA.

PubMed

Sembongi, Hiroshi; Di Re, Miriam; Bokori-Brown, Monika; Holt, Ian J

2007-10-01

Rearrangements of mitochondrial DNA (mtDNA) are a well-recognized cause of human disease; deletions are more frequent, but duplications are more readily transmitted to offspring. In theory, partial duplications of mtDNA can be resolved to partially deleted and wild-type (WT) molecules, via homologous recombination. Therefore, the yeast CCE1 gene, encoding a Holliday junction resolvase, was introduced into cells carrying partially duplicated or partially triplicated mtDNA. Some cell lines carrying the CCE1 gene had substantial amounts of WT mtDNA suggesting that the enzyme can mediate intramolecular recombination in human mitochondria. However, high levels of expression of CCE1 frequently led to mtDNA loss, and so it is necessary to strictly regulate the expression of CCE1 in human cells to ensure the selection and maintenance of WT mtDNA.

Spent coffee ground as a new bulking agent for accelerated biodrying of dewatered sludge.

PubMed

Hao, Zongdi; Yang, Benqin; Jahng, Deokjin

2018-07-01

The feasibility of using spent coffee ground (SCG) as a new bulking agent for biodrying of dewatered sludge (DS) was investigated in comparison with two other frequently-used bulking agents, air-dried sludge (AS) and sawdust (SD). Results showed that the moisture contents (MC) of 16-day DS biodrying with AS (Trial A), SCG (Trial B) and SD (Trial C) decreased from 70.14 wt%, 68.25 wt% and 71.63 wt% to 59.12 wt%, 41.35 wt% and 57.69 wt%, respectively. In case of Trial B, the MC rapidly decreased to 46.16 wt% with the highest water removal (70.87%) within 8 days because of the longest high-temperature period (5.8 days). Further studies indicated that the abundant biodegradable volatile solids (BVS) and high dissolved organic matter (DOM) contents in SCG were the main driving forces for water removal. According to pyrosequencing data, Firmicutes, most of which were recognized as thermophiles, was rapidly enriched on Day 8 and became the dominant phylum in Trial B. Four thermophilic genera, Bacillus, Ureibacillus, Geobacillus and Thermobifida, which can produce thermostable hydrolytic extracellular enzymes, were the most abundant in Trial B, indicating that these thermophilic bacteria evolved during the long high-temperature period enhanced the biodegradation of BVS in SCG. The 8-day biodried product of Trial B was demonstrated to be an excellent solid fuel with low heating value (LHV) of 9284 kJ kg -1 , which was 2.1 and 1.8 times those of biodried products with AS and SD, respectively. Thus SCG was found to be an excellent bulking agent accelerating DS biodrying and producing a solid fuel with a high calorific value. Copyright © 2018 Elsevier Ltd. All rights reserved.

Spi-C has opposing effects to PU.1 on gene expression in progenitor B cells.

PubMed

Schweitzer, Brock L; Huang, Kelly J; Kamath, Meghana B; Emelyanov, Alexander V; Birshtein, Barbara K; DeKoter, Rodney P

2006-08-15

The Ets transcription factor Spi-C, expressed in B cells and macrophages, is closely related to PU.1 and has the ability to recognize the same DNA consensus sequence. However, the function of Spi-C has yet to be determined. The purpose of this study is to further examine Spi-C activity in B cell development. First, using retroviral vectors to infect PU.1(-/-) fetal liver progenitors, Spi-C was found to be inefficient at inducing cytokine-dependent proliferation and differentiation of progenitor B (pro-B) cells or macrophages relative to PU.1 or Spi-B. Next, Spi-C was ectopically expressed in fetal liver-derived, IL-7-dependent pro-B cell lines. Wild-type (WT) pro-B cells ectopically expressing Spi-C (WT-Spi-C) have several phenotypic characteristics of pre-B cells such as increased CD25 and decreased c-Kit surface expression. In addition, WT-Spi-C pro-B cells express increased levels of IgH sterile transcripts and reduced levels of expression and transcription of the FcgammaRIIb gene. Gel-shift analysis suggests that Spi-C, ectopically expressed in pro-B cells, can bind PU.1 consensus sites in the IgH intronic enhancer and FcgammaRIIb promoter. Transient transfection analysis demonstrated that PU.1 functions to repress the IgH intronic enhancer and activate the FcgammaRIIb promoter, while Spi-C opposes these activities. WT-Spi-C pro-B cells have reduced levels of dimethylation on lysine 9 of histone H3 within the IgH 3' regulatory region, indicating that Spi-C can contribute to removal of repressive features in the IgH locus. Overall, these studies suggest that Spi-C may promote B cell differentiation by modulating the activity of PU.1-dependent genes.

CD205-TLR9-IL-12 axis contributes to CpG-induced oversensitive liver injury in HBsAg transgenic mice by promoting the interaction of NKT cells with Kupffer cells.

PubMed

Hou, Xin; Hao, Xiaolei; Zheng, Meijuan; Xu, Congfei; Wang, Jun; Zhou, Rongbin; Tian, Zhigang

2017-08-01

Gut-derived bacterial products contribute to liver inflammation and injury during chronic hepatitis B virus infection; however, the underlying mechanisms remain obscure. In this study, hepatitis B surface antigen transgenic (HBs-Tg) mice and their wild-type (WT) control C57BL/6 mice were injected with CpG-oligodeoxynucleotides (ODNs) to mimic the translocation of gut microbial products into the systemic circulation. We found that, compared with the WT mice, the HBs-Tg mice were oversensitive to CpG-ODN-induced liver injury, which was dependent on natural killer T (NKT) cells. CpG-ODN injection enhanced the expression of Fas ligand (FasL) on NKT cells. In addition, hepatocytes from the HBs-Tg mice expressed higher levels of Fas than did those from the WT mice, which was further augmented by CpG-ODN. Interaction of Fas and FasL was involved in the cytotoxicity of NKT cells against hepatocytes in the HBs-Tg mice. Moreover, Kupffer cells in the HBs-Tg mice expressed higher levels of CD205 and produced greater amounts of interleukin (IL)-12 than did those in the WT mice. Finally, the depletion of Kupffer cells, neutralization of IL-12 or specific silencing of CD205 on Kupffer cells significantly inhibited CpG-ODN-induced liver injury and NKT activation in the HBs-Tg mice. Our data suggest that CD205-expressing Kupffer cells respond to CpG-ODNs and subsequently release IL-12 to promote NKT cell activation. Activated NKT cells induce liver damage through the Fas signaling pathway in HBs-Tg mice.

CD205-TLR9-IL-12 axis contributes to CpG-induced oversensitive liver injury in HBsAg transgenic mice by promoting the interaction of NKT cells with Kupffer cells

PubMed Central

Hou, Xin; Hao, Xiaolei; Zheng, Meijuan; Xu, Congfei; Wang, Jun; Zhou, Rongbin; Tian, Zhigang

2017-01-01

Gut-derived bacterial products contribute to liver inflammation and injury during chronic hepatitis B virus infection; however, the underlying mechanisms remain obscure. In this study, hepatitis B surface antigen transgenic (HBs-Tg) mice and their wild-type (WT) control C57BL/6 mice were injected with CpG-oligodeoxynucleotides (ODNs) to mimic the translocation of gut microbial products into the systemic circulation. We found that, compared with the WT mice, the HBs-Tg mice were oversensitive to CpG-ODN-induced liver injury, which was dependent on natural killer T (NKT) cells. CpG-ODN injection enhanced the expression of Fas ligand (FasL) on NKT cells. In addition, hepatocytes from the HBs-Tg mice expressed higher levels of Fas than did those from the WT mice, which was further augmented by CpG-ODN. Interaction of Fas and FasL was involved in the cytotoxicity of NKT cells against hepatocytes in the HBs-Tg mice. Moreover, Kupffer cells in the HBs-Tg mice expressed higher levels of CD205 and produced greater amounts of interleukin (IL)-12 than did those in the WT mice. Finally, the depletion of Kupffer cells, neutralization of IL-12 or specific silencing of CD205 on Kupffer cells significantly inhibited CpG-ODN-induced liver injury and NKT activation in the HBs-Tg mice. Our data suggest that CD205-expressing Kupffer cells respond to CpG-ODNs and subsequently release IL-12 to promote NKT cell activation. Activated NKT cells induce liver damage through the Fas signaling pathway in HBs-Tg mice. PMID:27041637

The effect of acidification of liquid whey protein concentrate on the flavor of spray-dried powder.

PubMed

Park, Curtis W; Bastian, Eric; Farkas, Brian; Drake, MaryAnne

2014-07-01

Off-flavors in whey protein negatively influence consumer acceptance of whey protein ingredient applications. Clear acidic beverages are a common application of whey protein, and recent studies have demonstrated that beverage processing steps, including acidification, enhance off-flavor production from whey protein. The objective of this study was to determine the effect of preacidification of liquid ultrafiltered whey protein concentrate (WPC) before spray drying on flavor of dried WPC. Two experiments were performed to achieve the objective. In both experiments, Cheddar cheese whey was manufactured, fat-separated, pasteurized, bleached (250 mg/kg of hydrogen peroxide), and ultrafiltered (UF) to obtain liquid WPC that was 13% solids (wt/wt) and 80% protein on a solids basis. In experiment 1, the liquid retentate was then acidified using a blend of phosphoric and citric acids to the following pH values: no acidification (control; pH 6.5), pH 5.5, or pH 3.5. The UF permeate was used to normalize the protein concentration of each treatment. The retentates were then spray dried. In experiment 2, 150 μg/kg of deuterated hexanal (D₁₂-hexanal) was added to each treatment, followed by acidification and spray drying. Both experiments were replicated 3 times. Flavor properties of the spray-dried WPC were evaluated by sensory and instrumental analyses in experiment 1 and by instrumental analysis in experiment 2. Preacidification to pH 3.5 resulted in decreased cardboard flavor and aroma intensities and an increase in soapy flavor, with decreased concentrations of hexanal, heptanal, nonanal, decanal, dimethyl disulfide, and dimethyl trisulfide compared with spray drying at pH 6.5 or 5.5. Adjustment to pH 5.5 before spray drying increased cabbage flavor and increased concentrations of nonanal at evaluation pH values of 3.5 and 5.5 and dimethyl trisulfide at all evaluation pH values. In general, the flavor effects of preacidification were consistent regardless of the pH to which the solutions were adjusted after spray drying. Preacidification to pH 3.5 increased recovery of D₁₂-hexanal in liquid WPC and decreased recovery of D₁₂-hexanal in the resulting powder when evaluated at pH 6.5 or 5.5. These results demonstrate that acidification of liquid WPC80 to pH 3.5 before spray drying decreases off-flavors in spray-dried WPC and suggest that the mechanism for off-flavor reduction is the decreased protein interactions with volatile compounds at low pH in liquid WPC or the increased interactions between protein and volatile compounds in the resulting powder. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Fabrication of Chitosan Silk-based Tracheal Scaffold Using Freeze-Casting Method

PubMed Central

Nematollahi, Zeinab; Tafazzoli-Shadpour, Mohammad; Zamanian, Ali; Seyedsalehi, Amir; Mohammad-Behgam, Shadmehr; Ghorbani, Fariba; Mirahmadi, Fereshte

2017-01-01

Background: Since the treatments of long tracheal lesions are associated with some limitations, tissue engineered trachea is considered as an alternative option. This study aimed at preparing a composite scaffold, based on natural and synthetic materials for tracheal tissue engineering. Methods: Nine chitosan silk-based scaffolds were fabricated using three freezing rates (0.5, 1, and 2°C/min) and glutaraldehyde (GA) concentrations (0, 0.4, and 0.8 wt%). Samples were characterized, and scaffolds having mechanical properties compatible with those of human trachea and proper biodegradability were selected for chondrocyte cell seeding and subsequent biological assessments. Results: The pore sizes were highly influenced by the freezing rate and varied from 135.3×372.1 to 37.8×83.4 µm. Swelling and biodegradability behaviors were more affected by GA rather than freezing rate. Tensile strength raised from 120 kPa to 350 kPa by an increment of freezing rate and GA concentration. In addition, marked stiffening was demonstrated by increasing elastic modulus from 1.5 MPa to 12.2 MPa. Samples having 1 and 2°C/min of freezing rate and 0.8 wt% GA concentration made a non-toxic, porous structure with tensile strength and elastic modulus in the range of human trachea, facilitating the chondrocyte proliferation. The results of 21-day cell culture indicated that glycosaminoglycans content was significantly higher for the rate of 2°C/min (12.04 µg/min) rather than the other (9.6 µg/min). Conclusion: A homogenous porous structure was created by freeze drying. This allows the fabrication of a chitosan silk scaffold cross-linked by GA for cartilage tissue regeneration with application in tracheal regeneration. PMID:28131109

Preparation and properties of high storage stability polyester polyol dispersion for two-component waterborne polyurethane coating

NASA Astrophysics Data System (ADS)

Hao, H.; Hu, J. Q.; Wang, F.; Tu, W. P.

2017-01-01

A new type of polyester polyol dispersion with good storage stability was prepared based on a hydrophilic monomer 5-sodium sulfodimethyl isophthalate (5-SIPM), and frequently-used monomers such as neopentyl glycol (NPG), dimethyl terephthalate (DMT), dimethyl phthalate (DMP) and trimethylolpropane (TMP) by the transpolycondensation and polycondensation method. The polyester polyol dispersion was characterized by FTIR and GPC. The proper content of these monomers were determined by the performance of polyester dispersion: the content of TMP was 15wt%, the content of NPG was 7.5wt% and the hydrophilic monomer 5-SIPM content was 5wt%. Two-component waterborne polyurethane (2K-WPU) coatings were prepared by Bayhydur® XP2487/1 and polyester polyol dispersions, which were stored before and after at 40 ° for 6 weeks, the prepared films have no differences in drying time, adhesion, pencil hardness, gloss and chemical resistance, the result also reveals that the polyester polyol dispersion have excellent storage stability resistance.

New-Generation Aluminum Composite with Bottom Ash Industrial Waste

NASA Astrophysics Data System (ADS)

Mandal, A. K.; Sinha, O. P.

2018-02-01

Industrial waste bottom ash (BA) from a pulverized coal combustion boiler containing hard wear-resistant particles was utilized in this study to form an aluminum composite through a liquid metallurgy route. Composites comprising 5 wt.% and 10 wt.% bottom ash were characterized for their physiochemical, microstructural, mechanical, as well as tribological properties, along with pure aluminum. Scanning electron microscopy (SEM) microstructure revealed uniform distribution of BA particles throughout the matrix of the composite, whereas x-ray diffraction (XRD) analysis confirmed presence of aluminosilicate phase. Addition of 10 wt.% BA improved the Brinell hardness number (BHN) from 13 to 19 and ultimate tensile strength (UTS) from 71 MPa to 87 MPa, whereas ductility was adversely reduced after 5% BA addition. Incorporation of BA particles resulted in reduced dry sliding wear rates examined up to 80 N load compared with aluminum. Hence, such composites having lower cost could be applied as significantly hard, wear-resistant materials in applications in the automotive industry.

New-Generation Aluminum Composite with Bottom Ash Industrial Waste

NASA Astrophysics Data System (ADS)

Mandal, A. K.; Sinha, O. P.

2018-06-01

Industrial waste bottom ash (BA) from a pulverized coal combustion boiler containing hard wear-resistant particles was utilized in this study to form an aluminum composite through a liquid metallurgy route. Composites comprising 5 wt.% and 10 wt.% bottom ash were characterized for their physiochemical, microstructural, mechanical, as well as tribological properties, along with pure aluminum. Scanning electron microscopy (SEM) microstructure revealed uniform distribution of BA particles throughout the matrix of the composite, whereas x-ray diffraction (XRD) analysis confirmed presence of aluminosilicate phase. Addition of 10 wt.% BA improved the Brinell hardness number (BHN) from 13 to 19 and ultimate tensile strength (UTS) from 71 MPa to 87 MPa, whereas ductility was adversely reduced after 5% BA addition. Incorporation of BA particles resulted in reduced dry sliding wear rates examined up to 80 N load compared with aluminum. Hence, such composites having lower cost could be applied as significantly hard, wear-resistant materials in applications in the automotive industry.

Mislocalization of SLP-76 leads to aberrant inflammatory cytokine and autoantibody production.

PubMed

Sonnenberg, Gregory F; Mangan, Paul R; Bezman, Natalie A; Sekiguchi, Debora R; Luning Prak, Eline T; Erikson, Jan; Maltzman, Jonathan S; Jordan, Martha S; Koretzky, Gary A

2010-03-18

Central and peripheral tolerance is required to prevent immune responses to self-antigens. We now present a mouse model in which wild-type (WT) SH2 domain-containing leukocyte phosphoprotein of 76 kDa (SLP-76) has been constitutively targeted to the membrane, where CD4+ T cells become spontaneously dysregulated and develop an inflammatory phenotype. Mice bearing membrane-targeted SLP-76 (MTS) have a partial T-cell lymphopenia and impaired signaling though the mature T-cell receptor. The CD4+ T cells that develop in these mice possess an activated-like phenotype and are skewed toward the inflammatory T(H)1 and T(H)17 lineages. MTS mice also spontaneously develop autoantibodies at an early age. To rule out abnormal thymic selection as the sole cause of the MTS phenotype, we expressed WT SLP-76 along with the MTS followed by deletion of the WT allele in peripheral T cells. The peripheral MTS-expressing T cells demonstrate skewed cytokine responses when transferred into lymphopenic hosts. Thus, the abnormal effector T-cell phenotype still occurs in the presence of preserved central and peripheral tolerance, suggesting that diminished T-cell receptor signaling can promote skewed T-cell responses.

Claudin 5 Expression in Mouse Seminiferous Epithelium Is Dependent upon the Transcription Factor Ets Variant 5 and Contributes to Blood-Testis Barrier Function1

PubMed Central

Morrow, Carla M.K.; Tyagi, Gaurav; Simon, Liz; Carnes, Kay; Murphy, Kenneth M.; Cooke, Paul S.; Hofmann, Marie-Claude C.; Hess, Rex A.

2009-01-01

The blood-testis barrier (BTB) is formed by tight junctions between Sertoli cells. Results of previous studies suggested that the barrier is deficient in ets variant 5 (ETV5) gene-deleted mice; therefore, microarray data were examined for changes in tight junction-associated genes. The tight junctional protein claudin 5 (CLDN5) was decreased in testes of 8-day-old Etv5−/− pups. The study reported herein examined the expression of CLDN5 in wild-type (WT) and Etv5−/− mice and evaluated its contribution to BTB function. CLDN5 protein expression was evaluated in 8-day-old WT and Etv5−/− and adult WT, Etv5−/−, and W/Wv testes by immunohistochemistry and in 8-day-old WT Sertoli cell-enriched and germ cell-enriched fractions by immunocytochemistry. Cldn5 mRNA expression was evaluated in 0- to 20-day-old and adult WT mice and in 8-day-old and adult Etv5−/− mice via quantitative PCR. Tracer studies were performed in adult WT, Etv5−/−, and W/Wv mice. The results indicate the following: 1) CLDN5 was expressed in Sertoli cells, spermatogonia, and preleptotene spermatocytes. 2) Seminiferous epithelial CLDN5 expression depended upon both the presence of germ cells and ETV5. 3) CLDN5 expression in testicular vascular endothelium and rete testis epithelium was ETV5 independent. 4) Cldn5 mRNA expression increased in the testes of juvenile mice at the time of BTB formation. 5) Testes of Etv5−/− and W/Wv mice, which are both deficient in seminiferous epithelial CLDN5 expression, had biotin tracer leakage from the interstitial space into the seminiferous tubule lumen. In conclusion, CLDN5 is expressed in the seminiferous epithelium, appears to be regulated by multiple influences, and contributes to BTB function. PMID:19571261

In Situ Growth of Metal Sulfide Nanocrystals in Poly(3-hexylthiophene): [6,6]-Phenyl C61-Butyric Acid Methyl Ester Films for Inverted Hybrid Solar Cells with Enhanced Photocurrent.

PubMed

Yang, Chunyan; Sun, Yingying; Li, Xinjie; Li, Cheng; Tong, Junfeng; Li, Jianfeng; Zhang, Peng; Xia, Yangjun

2018-06-20

It has been reported that the performance of bulk heterojunction organic solar cells can be improved by incorporation of nano-heterostructures of metals, semiconductors, and dielectric materials in the active layer. In this manuscript, CdS or Sb 2 S 3 nanocrystals were in situ generated inside the poly(3-hexylthiophene): [6,6]-phenyl C61-butyric acid (P3HT:PC 61 BM) system by randomly mixing P3HT and PC 61 BM in the presence of cadmium or antimony xanthate precursor. Hybrid solar cells (HSCs) with the configurations of tin-doped indium oxide substrate (ITO)/CdS interface layer/P3HT:PC 61 BM: x wt.% CdS/MoO 3 /Ag and ITO/CdS interface layer /P3HT:PC 61 BM: x wt.% Sb 2 S 3 /MoO 3 /Ag were fabricated. Hybrid active layers (P3HT:PC 61 BM: x wt.% CdS or P3HT:PC 61 BM: x wt.% Sb 2 S 3 ) were formed completely by thermally annealing the film resulting in the decomposition of the cadmium or antimony xanthate precursor to CdS or Sb 2 S 3 nanocrystals, respectively. The effects of x wt.% CdS (or Sb 2 S 3 ) nanocrystals on the performance of the HSCs were studied. From UV-Vis absorption, hole mobilities, and surface morphological characterizations, it has been proved that incorporation of 3 wt.% CdS (or Sb 2 S 3 ) nanocrystals in the active layer of P3HT:PC 61 BM-based solar cells improved the optical absorption, the hole mobility, and surface roughness in comparison with P3HT:PC 61 BM-based solar cells, thus resulting in the improved power conversion efficiencies (PCEs) of the devices.

Characterization of mutant tobacco mosaic virus coat protein that interferes with virus cell-to-cell movement.

PubMed

Bendahmane, Mohammed; Szecsi, Judit; Chen, Iju; Berg, R Howard; Beachy, Roger N

2002-03-19

Expression of tobacco mosaic virus (TMV) coat protein (CP) in plants confers resistance to infection by TMV and related tobamoviruses. Certain mutants of the CP (CP(T42W)) provide much greater levels of resistance than wild-type (wt) CP. In the present work, infection induced by RNA transcripts of TMV clones that contain wt CP or mutant CP(T42W) fused to the green fluorescent protein (GFP) (TMV-CP:GFP, TMV-CP(T42W):GFP) and clones harboring TMV movement protein (MP):GFP were followed in nontransgenic and transgenic tobacco BY-2 protoplasts and Nicotiana tabaccum Xanthi-nn plants that express wt CP or CP(T42W). On nontransgenic and wt CP transgenic plants, TMV-CP:GFP produced expanding, highly fluorescent disk-shaped areas. On plants expressing CP(T42W), infection by TMV-CP:GFP or TMV-MP:GFP-CP produced infection sites of smaller size that were characterized by low fluorescence, reflecting reduced levels of virus spread and reduced accumulation of both CP:GFP and MP:GFP. TMV-CP(T42W):GFP failed to produce visible infection sites on nontransgenic plants, yet produced normal infection sites on MP-transgenic plants that produce MP. TMV infection of transgenic BY-CP(T42W) protoplasts resulted in very low levels of MP accumulation, whereas on BY-CP protoplasts (containing wt CP), infection produced higher levels of MP than in nontransgenic BY-2 cells. The results suggest that wt CP has a positive effect on the production of MP, whereas the CP(T42W) has a negative effect on MP accumulation and/or function. This effect results in very high levels of resistance to TMV infection in plants containing CP(T42W). This report shows that the CP of a plant virus regulates production of the MP, and that a mutant CP interferes with MP accumulation and cell-to-cell movement of infection.

Influence of temperature on fluoride toxicity and bioaccumulation in the nonindigenous freshwater mollusk Dreissena polymorpha Pallas, 1769.

PubMed

Del Piero, Stefania; Masiero, Luciano; Casellato, Sandra

2012-11-01

Fluoride toxicity and bioaccumulation tests (short- and long-term) were performed on the nonindigenous freshwater mollusk Dreissena polymorpha at two different temperatures: 17 ± 0.5°C and 22 ± 0.5°C. Concentrations that did not result in toxicity in short-term experiments (96 h) induced effects over a longer period (17 weeks), especially at the warmest temperature, highlighting the role of this parameter. Fluoride bioaccumulation increased linearly with increasing concentration and temperature, reaching 4,202 µg F(-)/g dry weight in soft tissues only after 48 h of exposure at 22°C at a concentration of 640 mg F(-)/L. Comparing tolerance to fluoride and bioaccumulation values of this species with those of other freshwater invertebrates, D. polymorpha was much more resistant and revealed its capacity to accumulate a great quantity of this xenobiotic substance. The results of the present study demonstrated that fluoride accumulation in the soft tissue of this animal was much higher (up to 1,409.6 µg F(-)/g dry wt) than that in its shell (up to 706.4 µg F(-)/g dry wt). If we consider this datum and the fact that D. polymorpha is widespread in many aquatic ecosystems around the world, representing a food source for many birds and other vertebrates, we must acknowledge the possibility that it can represent a serious danger in view of fluoride biomagnification in the aquatic environment. Copyright © 2012 SETAC.

The effect of calorie restriction on the presence of apoptotic ovarian cells in normal wild type mice and low-plasma-IGF-1 Laron dwarf mice

PubMed Central

2013-01-01

Background It is known that caloric restriction extends lifespan and can minimize age-related dysfunction of the reproductive system. We became interested in how caloric restriction influences apoptosis, which is a crucial process that maintains ovarian cell homeostasis. Methods We examined ovarian cells in: 2.5-year-old wild type mice on caloric restriction (CR) or fed ad libitum (AL) and Laron dwarf mice (GHR-KO) at the same ages on CR or fed AL. Apoptosis was assessed by histochemical analysis on paraffin sections of ovarian tissue. Results Morphological and histochemical analysis revealed that CR improved reproductive potential in 2.5-year-old WT littermates and GHR-KO female mice, as indicated by the increased number of ovarian follicles. The level of apoptosis in ovarian tissue was higher in WT mice on a CR diet compared with WT mice on the AL diet. In GHR-KO mice, the level of apoptosis in ovaries was similar for mice on CR and on AL diets and bigger than in WT mice on CR. Conclusions Morphological and histochemical analysis revealed a younger biological age of the ovaries in 2-year-old WT littermates and GHR-KO female mice on CR compared with animals fed AL. PMID:24063422

Characterization of Cross-Linked Porous Gelatin Carriers and Their Interaction with Corneal Endothelium: Biopolymer Concentration Effect

PubMed Central

Lai, Jui-Yang; Ma, David Hui-Kang; Lai, Meng-Heng; Li, Ya-Ting; Chang, Ren-Jie; Chen, Li-Mei

2013-01-01

Cell sheet-mediated tissue regeneration is a promising approach for corneal reconstruction. However, the fragility of bioengineered corneal endothelial cell (CEC) monolayers allows us to take advantage of cross-linked porous gelatin hydrogels as cell sheet carriers for intraocular delivery. The aim of this study was to further investigate the effects of biopolymer concentrations (5–15 wt%) on the characteristic and safety of hydrogel discs fabricated by a simple stirring process combined with freeze-drying method. Results of scanning electron microscopy, porosity measurements, and ninhydrin assays showed that, with increasing solid content, the pore size, porosity, and cross-linking index of carbodiimide treated samples significantly decreased from 508±30 to 292±42 µm, 59.8±1.1 to 33.2±1.9%, and 56.2±1.6 to 34.3±1.8%, respectively. The variation in biopolymer concentrations and degrees of cross-linking greatly affects the Young’s modulus and swelling ratio of the gelatin carriers. Differential scanning calorimetry measurements and glucose permeation studies indicated that for the samples with a highest solid content, the highest pore wall thickness and the lowest fraction of mobile water may inhibit solute transport. When the biopolymer concentration is in the range of 5–10 wt%, the hydrogels have high freezable water content (0.89–0.93) and concentration of permeated glucose (591.3–615.5 µg/ml). These features are beneficial to the in vitro cultivation of CECs without limiting proliferation and changing expression of ion channel and pump genes such as ATP1A1, VDAC2, and AQP1. In vivo studies by analyzing the rabbit CEC morphology and count also demonstrate that the implanted gelatin discs with the highest solid content may cause unfavorable tissue-material interactions. It is concluded that the characteristics of cross-linked porous gelatin hydrogel carriers and their triggered biological responses are in relation to biopolymer concentration effects. PMID:23382866

Cultivation, characterization, and properties of Chlorella vulgaris microalgae with different lipid contents and effect on fast pyrolysis oil composition.

PubMed

Adamakis, Ioannis-Dimosthenis; Lazaridis, Polykarpos A; Terzopoulou, Evangelia; Torofias, Stylianos; Valari, Maria; Kalaitzi, Photeini; Rousonikolos, Vasilis; Gkoutzikostas, Dimitris; Zouboulis, Anastasios; Zalidis, Georgios; Triantafyllidis, Konstantinos S

2018-06-01

A systematic study of the effect of nitrogen levels in the cultivation medium of Chlorella vulgaris microalgae grown in photobioreactor (PBR) on biomass productivity, biochemical and elemental composition, fatty acid profile, heating value (HHV), and composition of the algae-derived fast pyrolysis (bio-oil) is presented in this work. A relatively high biomass productivity and cell concentration (1.5 g of dry biomass per liter of cultivation medium and 120 × 10 6 cells/ml, respectively) were achieved after 30 h of cultivation under N-rich medium. On the other hand, the highest lipid content (ca. 36 wt.% on dry biomass) was obtained under N-depletion cultivation conditions. The medium and low N levels favored also the increased concentration of the saturated and mono-unsaturated C16:0 and C18:1(n-9) fatty acids (FA) in the lipid/oil fraction, thus providing a raw lipid feedstock that can be more efficiently converted to high-quality biodiesel or green diesel (via hydrotreatment). In terms of overall lipid productivity, taking in consideration both the biomass concentration in the medium and the content of lipids on dry biomass, the most effective system was the N-rich one. The thermal (non-catalytic) pyrolysis of Chlorella vulgaris microalgae produced a highly complex bio-oil composition, including fatty acids, phenolics, ethers, ketones, etc., as well as aromatics, alkanes, and nitrogen compounds (pyrroles and amides), originating from the lipid, protein, and carbohydrate fractions of the microalgae. However, the catalytic fast pyrolysis using a highly acidic ZSM-5 zeolite, afforded a bio-oil enriched in mono-aromatics (BTX), reducing at the same time significantly oxygenated compounds such as phenolics, acids, ethers, and ketones. These effects were even more pronounced in the catalytic fast pyrolysis of Chlorella vulgaris residual biomass (after extraction of lipids), thus showing for the first time the potential of transforming this low value by-product towards high added value platform chemicals.

IL-15-deficient mice develop enhanced allergic responses to airway allergen exposure

PubMed Central

Mathias, Clinton B.; Schramm, Craig M.; Guernsey, Linda A.; Wu, Carol A.; Polukort, Stephanie H.; Rovatti, Jeffrey; Ser-Dolansky, Jennifer; Secor, Eric; Schneider, Sallie S.; Thrall, Roger S.; Aguila, Hector L.

2017-01-01

Background Interleukin-15 is a pleiotropic cytokine that is critical for the development and survival of multiple hematopoietic lineages. Mice lacking IL-15 have selective defects in populations of several pro-allergic immune cells including natural killer (NK) cells, NKT cells, and memory CD8+T cells. We therefore hypothesized that IL-15−/− mice will have reduced inflammatory responses during the development of allergic airway disease (AAD). Objective To determine whether IL-15−/− mice have attenuated allergic responses in a mouse model of AAD. Methods C57BL/6 wild-type (WT) and IL-15−/− mice were sensitized and challenged with ovalbumin (OVA) and the development of AAD was ascertained by examining changes in airway inflammatory responses, Th2 responses, and lung histopathology. Results Here we report that IL-15−/− mice developed enhanced allergic responses in an OVA-induced model of AAD. In the absence of IL-15, OVA-challenged mice exhibited enhanced bronchial eosinophilic inflammation, elevated IL-13 production, and severe lung histopathology in comparison with WT mice. In addition, increased numbers of CD4+T and B cells in the spleens and broncholaveolar lavage (BAL) were also observed. Examination of OVA-challenged IL-15Rα−/− animals revealed a similar phenotype resulting in enhanced airway eosinophilia compared to WT mice. Adoptive transfer of splenic CD8+T cells from OVA-sensitized WT mice suppressed the enhancement of eosinophilia in IL-15−/− animals to levels observed in WT mice, but had no further effects. Conclusion and Clinical Relevance These data demonstrate that mice with an endogenous IL-15 deficiency are susceptible to the development of severe, enhanced Th2-mediated AAD, which can be regulated by CD8+T cells. Furthermore, the development of disease as well as allergen-specific Th2 responses occurs despite deficiencies in several IL-15-dependent cell types including NK, NKT, and γδ T cells, suggesting that these cells or their subsets are dispensable for the induction of AAD in IL-15-deficient mice. PMID:28093832

Tribo-performance of epoxy hybrid composites reinforced with carbon fibers and potassium titanate whiskers

NASA Astrophysics Data System (ADS)

Suresha, B.; Harshavardhan, B.; Ravishankar, R.

2018-04-01

The present investigation deals with the fabrication and characterization of epoxy reinforced with bidirectional carbon fiber mat (CF/Ep) and filled with 2.5, 5 and 7.5 wt% potassium titanate whiskers (PTw) composites. The effect of PTw loading on hardness, tensile properties and dry sliding wear behaviour of CF/Ep composite were carefully investigated in expectation of providing valuable information for the application of hybrid CF/Ep composites. Results indicated that the incorporation of PTw actually improved the hardness, tensile strength and tensile modulus of CF/Ep composites. Meanwhile, the specific wear rate of CF/Ep filled by 5 wt % PTw reached to 6.3× 10-14 m3/N-m, which is 41% lower than that of CF/Ep composite at the same dry sliding condition. It also seen that the fiber and filler worked synergistically to enhance the wear resistance. Further, for all composites the friction coefficient increases with increase in load and sliding velocity. However, PTw reinforced CF/Ep exhibited considerably higher coefficient of friction compared to unfilled ones, while PTw filler loading of 5 wt% was effective in reducing the specific wear rate of CF/Ep composite. The carbon fiber carried the applied load between the contact surfaces and protected the epoxy from severe abrasion of the counterface. At the same time, the exposed PTw out of the epoxy matrix around the fiber inhibited the direct scraping between the fiber and counterface so that the fibers could be less directly impacted during the subsequent wear process and they were protected from severe damage.

Potential of Staphylococcus xylosus strain for recovering nickel ions from aqueous solutions

NASA Astrophysics Data System (ADS)

Al-Gheethi, A.; Efaq, A. N.; Mohamed, R. M.; Abdel-Monem, M.; Amir Hashim, M.

2017-10-01

The potential of bacterial biomass for the biosorption of heavy metals has investigated extensively. However, the bacterial species exhibited different affinities toward the heavy metals ions based on their differences in cell wall characteristics, structure and physiological status (living or dead cells). In this study, the potential of living and dead cells of Staphylococcus xylosus 222W for removal nickel ions from aqueous solution as a function for physiological status, nickel and biomass concentrations, time, pH and temperature was investigated. The pre-treatment of bacterial cells was performed by the heating at 100 °C for 15 min. The removal experiments were conducted in the lab scale. The results revealed that the dead cells exhibited more efficiency in removing nickel ions than living cells at all investigated concentrations (2 to 10 mM). The biosorption efficiency (E %) increased with increasing in biomass cells to limit concentrations (0.1 to 1 g dry wt L-1). The maximum removal of nickel was 81.41 vs. 77.10 % by living and dead cells, respectively achieved after 9 and 10 hrs of the incubation period, respectively. The acidic conditions decrease the efficiency of metal removal, while the optimal removal was recorded at pH 8 for both biomass (living and dead cells). The maximum uptake capacity of S. xylosus 222W (living and dead cells) was recorded at 37oC, the percentage removed being 75.90 vs. 84.92 %, respectively. It can be concluded that S. xylosus 222W exhibited high potential and affinity to remove of nickel ions from aqueous solution.

Optimizing the design and in vitro evaluation of bioreactive glucose oxidase-microspheres for enhanced cytotoxicity against multidrug resistant breast cancer cells.

PubMed

Cheng, Ji; Liu, Qun; Shuhendler, Adam J; Rauth, Andrew M; Wu, Xiao Yu

2015-06-01

Glucose oxidase (GOX) encapsulated in alginate-chitosan microspheres (GOX-MS) was shown in our previous work to produce reactive oxygen species (ROS) in situ and exhibit anticancer effects in vitro and in vivo. The purpose of present work was to optimize the design and thus enhance the efficacy of GOX-MS against multidrug resistant (MDR) cancer cells. GOX-MS with different mean diameters of 4, 20 or 140 μm were prepared using an emulsification-internal gelation-adsorption-chitosan coating method with varying compositions and conditions. The GOX loading efficiency, loading level, relative bioactivity of GOX-MS, and GOX leakage were determined and optimal chitosan concentrations in the coating solution were identified. The influence of particle size on cellular uptake, ROS generation, cytotoxicity and their underlying mechanisms was investigated. At the same GOX dose and incubation time, smaller sized GOX-MS produced larger amounts of H2O2 in cell culture medium and greater cytotoxicity toward murine breast cancer MDR (EMT6/AR1.0) and wild type (EMT6/WT) cells. Fluorescence and confocal laser scanning microscopy revealed significant uptake of small sized (4 μm) GOX-MS by both MDR and WT cells, but no cellular uptake of large (140 μm) GOX-MS. The GOX-MS were equally effective in killing both MDR cells and WT cells. The cytotoxicity of the GOX formulations was positively correlated with membrane damage and lipid peroxidation. GOX-MS induced greater membrane damage and lipid peroxidation in MDR cells than the WT cells. These results suggest that the optimized, small micron-sized GOX-MS are highly effective against MDR breast cancer cells. Copyright © 2015 Elsevier B.V. All rights reserved.

p53-dependent cell death/apoptosis is required for a productive adenovirus infection.

PubMed

Hall, A R; Dix, B R; O'Carroll, S J; Braithwaite, A W

1998-09-01

The p53 tumor suppressor protein binds to both cellular and viral proteins, which influence its biological activity. One such protein is the large E1b tumor antigen (E1b58kDa) from adenoviruses (Ads), which abrogates the ability of p53 to transactivate various promoters. This inactivation of p53 function is believed to be the mechanism by which E1b58kDa contributes to the cell transformation process. Although the p53-E1b58kDa complex occurs during infection and is conserved among different serotypes, there are limited data demonstrating that it has a role in virus replication. However, loss of p53 expression occurs after adenovirus infection of human cells and an E1b58kDa deletion mutant (Onyx-015, also called dl 1520) selectively replicates in p53-defective cells. These (and other) data indicate a plausible hypothesis is that loss of p53 function may be conducive to efficient adenovirus replication. However, wild-type (wt) Ad5 grows more efficiently in cells expressing a wt p53 protein. These studies indicate that the hypothesis may be an oversimplification. Here, we show that cells expressing wt p53, as well as p53-defective cells, allow adenovirus replication, but only cells expressing wt p53 show evidence of virus-induced cytopathic effect. This correlates with the ability of adenovirus to induce cell death. Our data indicate that p53 plays a necessary part in mediating cellular destruction to allow a productive adenovirus infection. In contrast, p53-deficient cells are less sensitive to the cytolytic effects of adenovirus and as such raise questions about the use of E1b58kDa-deficient adenoviruses in tumor therapy.

Insights on the SO2 Poisoning of Pt3Co/VC and Pt/VC Fuel Cell Catalysts

DTIC Science & Technology

2010-01-01

catalyst is performed at the cathode of proton exchange membrane fuel cells ( PEMFCs ) in order to link previously reported results at the elec- trode...stripping voltammetry and underpotential deposition (upd) of copper adatoms. Then the performance of PEMFC cathodes employing 30wt.% Pt3Co/VC and 50wt.% Pt/VC...proton exchange membrane fuel cells( PEMFCs )in order to link previously reported results at the elec- trode/solution interface to the FC environment. First

Rheological modification of corn stover biomass at high solids concentrations

Treesearch

Joseph R. Samaniuk; C. Tim Scott; Thatcher W. Root; Daniel J. Klingenberg

2012-01-01

Additives were tested for their ability to modify the rheology of lignocellulosic biomass. Additive types included water-soluble polymers (WSPs), surfactants, and fine particles. WSPs were the most effective rheological modifiers, reducing yield stresses of concentrated biomass by 60–80% for additive concentrations of 1–2 wt. % (based on mass of dry biomass solids)....

Neuroelectric Predictors of Performance Accuracy.

DTIC Science & Technology

1986-07-31

3OM REz),S evn-eae Ialca wav IM EOmS, avrae Iro seemujcsmrentsgiiatl ifrnoAILR RIL F& the comarso oflf-cuae()wt etiacrt B codiios or ritacre...man. CE jn N~L~bi... ~,266. Posner. M. (1978) CbhrOnmIric C2QUt1dri Q1~ jIQ MidI Erlbaum: New Jersey. Rohrbaugh. J.W., Syndulko, K., Sanquist, T.F

Infiltration of Matrix-Non-producers Weakens the Salmonella Biofilm and Impairs Its Antimicrobial Tolerance and Pathogenicity.

PubMed

Srinandan, Chakravarthy S; Elango, Monalisha; Gnanadhas, Divya P; Chakravortty, Dipshikha

2015-01-01

Bacterial biofilms display a collective lifestyle, wherein the cells secrete extracellular polymeric substances (EPS) that helps in adhesion, aggregation, stability, and to protect the bacteria from antimicrobials. We asked whether the EPS could act as a public good for the biofilm and observed that infiltration of cells that do not produce matrix components weakened the biofilm of Salmonella enterica serovar Typhimurium. EPS production was costly for the producing cells, as indicated by a significant reduction in the fitness of wild type (WT) cells during competitive planktonic growth relative to the non-producers. Infiltration frequency of non-producers in the biofilm showed a concomitant decrease in overall productivity. It was apparent in the confocal images that the non-producing cells benefit from the EPS produced by the Wild Type (WT) to stay in the biofilm. The biofilm containing non-producing cells were more significantly susceptible to sodium hypochlorite and ciprofloxacin treatment than the WT biofilm. Biofilm infiltrated with non-producers delayed the pathogenesis, as tested in a murine model. The cell types were spatially assorted, with non-producers being edged out in the biofilm. However, cellulose was found to act as a barrier to keep the non-producers away from the WT microcolony. Our results show that the infiltration of non-cooperating cell types can substantially weaken the biofilm making it vulnerable to antibacterials and delay their pathogenesis. Cellulose, a component of EPS, was shown to play a pivotal role of acting as the main public good, and to edge-out the non-producers away from the cooperating microcolony.

Relationship between peat geochemistry and depositional environments, Cranberry Island, Maine

USGS Publications Warehouse

Raymond, R.; Cameron, C.C.; Cohen, A.D.

1987-01-01

The Heath, Great Cranberry Island, Maine, offers a unique locality for studying lateral and vertical relationships between radically different peat types within 1 km2. The majority of The Heath is a Sphagnum moss-dominated raised bog. Surrounding the raised bog is a swamp/marsh complex containing grass, sedge, Sphagnum moss, alder, tamarack, and skunk cabbage. Swamp/ marsh-deposited peat occurs both around the margins of The Heath and under Sphagnum-dominated peat, which was deposited within the raised bog. A third peat type, dominated by herbaceous aquatics, is present underlying the swamp/marsh-dominated peat but is not present as a dominant botanical community of The Heath. The three peat types have major differences in petrographic characteristics, ash contents, and associated minerals. Sulfur contents range from a low of 0.19 wt.% (dry) within the raised bog to a high of 4.44 wt% (dry) near the west end of The Heath, where swamp/marsh peat occurring directly behind a storm beach berm has been influenced by marine waters. The presence of major geochemical variations within a 1-km2 peat deposit suggests the need for in-depth characterization of potential peat resources prior to use. ?? 1987.

Loosely bound oxytetracycline in riverine sediments from two tributaries of the Chesapeake Bay

USGS Publications Warehouse

Simon, N.S.

2005-01-01

The fate of antibiotics that bind to riverine sediment is not well understood. A solution used in geochemical extraction schemes to determine loosely bound species in sediments, 1 M MgCl2 (pH 8), was chosen to determine loosely bound, and potentially bioavailable, tetracycline antibiotics (TCs), including oxytetracycline (5-OH tetracycline) (OTC) in sediment samples from two rivers on the eastern shore of the Chesapeake Bay. Bottom sediments were collected at sites upstream from, at, and downstream from municipal sewage-treatment plants (STPs) situated on two natural waterways, Yellow Bank Stream, MD, and the Pocomoke River, MD. Concentrations of easily desorbed OTC ranged from 0.6 to approximately 1.2 ??g g-1 dry wt sediment in Yellow Bank Stream and from 0.7 to approximately 3.3 ??g g-1 dry wt sediment in the Pocomoke River. Concentrations of easily desorbable OTC were generally smaller in sediment upstream than in sediment downstream from the STP in the Pocomoke River. STPs and poultry manure are both potential sources of OTC to these streams. OTC that is loosely bound to sediment is subject to desorption. Other researchers have found desorbed TCs to be biologically active compounds.

Production of Hexagenia limbata nymphs in contaminated sediments in the Upper Great Lakes connecting channels

USGS Publications Warehouse

Edsall, Thomas A.; Manny, Bruce A.; Schloesser, Donald W.; Nichols, Susan J.; Frank, Anthony M.

1991-01-01

In April through October 1986, we sampled sediments and populations of nymphs of the burrowing mayfly, Hexagenia limbata (Serville), at 11 locations throughout the connecting channels of the upper Great Lakes, to determine if sediment contaminants adversely affected nymph production. Production over this period was high (980 to 9231 mg dry wt m-2) at the five locations where measured sediment levels of oil, cyanide, and six metals were below the threshold criteria of the U.S. Environmental Protection Agency and the Ontario Ministry of Environment for contaminated or polluted sediments, and also where the criterion for visible oil given in the Water Quality Agreement between the U.S.A. and Canada for connecting waters of the Great Lakes was not exceeded. At the other six locations where sediments were polluted, production was markedly lower (359 to 872 mg dry wt m-2). This finding is significant because it indicates that existing sediment quality criteria can be applied to protect H. limbata from oil, cyanide, and metals in the Great Lakes and connecting channels where the species fulfills a major role in secondary production and trophic transfer of energy.

Palaeoenvironmental reconstruction of Hüsamlar coal seam, SW Turkey

NASA Astrophysics Data System (ADS)

Büçkün, Zeynep; İnaner, Hülya; Oskay, Riza Görkem; Christanis, Kimon

2015-06-01

The Ören and Yatağan Basins in SW Turkey host several Miocene coal deposits currently under exploitation for power generation. The present study aims to provide insight into the palaeoenvironmental conditions, which controlled the formation of the Hüsamlar coal seam located in Ören Basin. The coal seam displays many sharp alternations of matrix lignite beds and inorganic, lacustrine sediment layers. The coal is a medium-to-high ash lignite (10.47-31.16 wt%, on dry basis) with high total sulphur content (up to 10 wt%, on dry, ash-free basis), which makes it prone to self-combustion. The maceral composition indicates that the peat-forming vegetation consisted of both arboreal and herbaceous plants, with the latter being predominant in the upper part of the seam. Mica and feldspars contribute to the low part of the seam; carbonates are dominant in the upper part, whereas quartz and pyrite are present along the entire coal profile. The sudden transitions of the telmatic to the lacustrine regime and reverse is attributed to tectonic movements that controlled water table levels in the palaeomire, which affected surface runoff and hence, clastic deposition.

Robust Immobilized Amine CO 2 Sorbent Pellets Utilizing a Poly(Chloroprene) Polymer Binder and Fly Ash Additive

DOE PAGES

Wilfong, Walter Christopher; Kail, Brian W.; Howard, Bret H.; ...

2016-08-04

Pelletization of ca. 50 wt % amine/silica carbon dioxide sorbents was achieved with the novel combination of fly ash (FA) as a strength additive and hydrophobic poly(chloroprene) (PC) as a binder. The PC content and overall synthesis procedure of these materials were optimized to produce pellets, labeled as FA/E100-S_(20/80)_12.2, with the highest ball-mill attrition resistance (<0.5 wt % by fines, 24 h) and maximum CO 2 capture capacity of 1.78 mmol CO 2 g -1. The strength of the pellets was attributed to hydrogen-bonding of the relatively homogeneous PC network with the interlocked FA and BIAS particles (DRIFTS, SEM-EDS). Themore » low degradation of 3–4 % in the pellet's CO 2 capture capacity under both dry TGA (7.5 h) and practical fixed-bed (6.5 h dry; 4.5 h humid,≈5 vol % H 2O) CO 2 adsorption–desorption conditions highlights the pellet's excellent cyclic stability. These robust pellet characteristics make PC/FA/sorbent materials promising for commercial scale, point-source CO 2 capture.« less

Fabrication of Activated Rice Husk Charcoal by Slip Casting as a Hybrid Material for Water Filter Aid

NASA Astrophysics Data System (ADS)

Tuaprakone, T.; Wongphaet, N.; Wasanapiarnpong, T.

2011-04-01

Activated charcoal has been widely used as an odor absorbent in household and water purification industry. Filtration equipment for drinking water generally consists of four parts, which are microporous membrane (porous alumina ceramic or diatomite, or porous polymer), odor absorbent (activated carbon), hard water treatment (ion exchange resin), and UV irradiation. Ceramic filter aid is usually prepared by slip casting of alumina or diatomite. The membrane offers high flux, high porosity and maximum pore size does not exceed 0.3 μm. This study investigated the fabrication of hybrid activated charcoal tube for water filtration and odor absorption by slip casting. The suitable rice husk charcoal and water ratio was 48 to 52 wt% by weight with 1.5wt% (by dry basis) of CMC binder. The green rice husk charcoal bodies were dried and fired between 700-900 °C in reduction atmosphere. The resulting prepared slip in high speed porcelain pot for 60 min and sintered at 700 °C for 1 h showed the highest specific surface area as 174.95 m2/g. The characterizations of microstructure and pore size distribution as a function of particle size were investigated.

Transfer of (40)K, (238)U, (210)Pb, and (210)Po from soil to plant in various locations in south of Syria.

PubMed

Al-Masri, M S; Al-Akel, B; Nashawani, A; Amin, Y; Khalifa, K H; Al-Ain, F

2008-02-01

Transfer factors of (40)K, (238)U, (210)Pb, and (210)Po from soil to some agriculture crops in various locations in south of Syria (Dara'a and Assuwaydaa districts) have been determined. Soil and vegetable crops (green pepper, cucumber, tomato, and eggplant), legumes crops (lentil, chickpea, and broad bean), fruit trees (apple, grape, and olives) and cereals (barley and wheat) were collected and analyzed for (238)U, (210)Pb, and (210)Po. The results have shown that higher transfer factors (calculated as Bqkg(-1) dry wt. plant material per Bqkg(-1) dry wt. soil) for (210)Po, (210)Pb and (238)U were observed in vegetable leaves than fruits and cereals leaves; the highest values of transfer factor (TF) for (238)U were found to be 0.1 for straw of chickpea. Transfer factors for (210)Po varied between 2.8x10(-2) and 2 in fruits of eggplant and grain of barley, respectively. In addition, several parameters affecting transfer factors of the radionuclides were evaluated. The results can be considered as base values for TF of natural radionuclides in the region.

Postsynthetic Functionalization of Mg-MOF-74 with Tetraethylenepentamine: Structural Characterization and Enhanced CO2 Adsorption.

PubMed

Su, Xiao; Bromberg, Lev; Martis, Vladimir; Simeon, Fritz; Huq, Ashfia; Hatton, T Alan

2017-03-29

Postsynthetic functionalization of magnesium 2,5-dihydroxyterephthalate (Mg-MOF-74) with tetraethylenepentamine (TEPA) resulted in improved CO 2 adsorption performance under dry and humid conditions. XPS, elemental analysis, and neutron powder diffraction studies indicated that TEPA was incorporated throughout the MOF particle, although it coordinated preferentially with the unsaturated metal sites located in the immediate proximity to the surface. Neutron and X-ray powder diffraction analyses showed that the MOF structure was preserved after amine incorporation, with slight changes in the lattice parameters. The adsorption capacity of the functionalized amino-Mg-MOF-74 (TEPA-MOF) for CO 2 was as high as 26.9 wt % versus 23.4 wt % for the original MOF due to the extra binding sites provided by the multiunit amines. The degree of functionalization with the amines was found to be important in enhancing CO 2 adsorption, as the optimal surface coverage improved performance and stability under both pure CO 2 and CO 2 /H 2 O coadsorption, and with partially saturated surface coverage, optimal CO 2 capacity could be achieved under both wet and dry conditions by a synergistic binding of CO 2 to the amines as well as metal centers.

Graphene as transparent electrode in Si solar cells: A dry transfer method

NASA Astrophysics Data System (ADS)

Zang, Yuan; Li, Lian-bi; Chu, Qing; Pu, Hongbin; Hu, Jichao; Jin, Haili; Zhang, Yan

2018-06-01

This paper present an experimental study on a new dry transfer method of graphene applied as transparent conducting electrode in textured silicon solar cells. Raman spectra of dry exfoliated graphene indicated a monolayer graphene was transferred onto textured Si cells and the dry exfoliated graphene with better crystalline quality is attained. Photovoltaic result shows the short circuit current of the dry transferred graphene on texture Si solar cell has an increase of 28% compared to the wet transferred graphene. It proves the possibility of dry transferred graphene as transparent conducting electrode in textured Si solar cell applications.

From coin cells to 400 mAh pouch cells: Enhancing performance of high-capacity lithium-ion cells via modifications in electrode constitution and fabrication

NASA Astrophysics Data System (ADS)

Trask, Stephen E.; Li, Yan; Kubal, Joseph J.; Bettge, Martin; Polzin, Bryant J.; Zhu, Ye; Jansen, Andrew N.; Abraham, Daniel P.

2014-08-01

In this article we describe efforts to improve performance and cycle life of cells containing Li1.2Ni0.15Mn0.55Co0.1O2-based positive and graphite-based negative electrodes. Initial work to identify high-performing materials, compositions, fabrication variables, and cycling conditions is conducted in coin cells. The resulting information is then used for the preparation of double-sided electrodes, assembly of pouch cells, and electrochemical testing. We report the cycling performance of cells with electrodes prepared under various conditions. Our data indicate that cells with positive electrodes containing 92 wt.% Li1.2Ni0.15Mn0.55Co0.1O2, 4 wt.% carbons (no graphite), and 4 wt.% PVdF (92-4-4) show ∼20% capacity fade after 1000 cycles in the 2.5-4.4 V range, significantly better than our baseline cells that show the same fade after only 450 cycles. Our analyses indicate that the major contributors to cell energy fade are capacity loss and impedance rise. Therefore incorporating approaches that minimize capacity fade and impedance rise, such as electrode coatings and electrolyte additives, can significantly enhance calendar and cycle life of this promising cell chemistry.