Effect of oxygen supply on Monascus pigments and citrinin production in submerged fermentation.

PubMed

Yang, Jian; Chen, Qi; Wang, Weiping; Hu, Jiajun; Hu, Chuan

2015-05-01

The influence of oxygen supply on Monascus pigments and citrinin production by Monascus ruber HS.4000 in submerged fermentation was studied. For Monascus cultivation with high pigments and low citrinin production, the initial growth phase, mid-stage phase, and later-stage production phase were separated by shifting oxygen supply. The optimal condition for the fermentation process in shake-flask fermentation was a three-stage rotating rate controlled strategy (0-48 h at 150 rpm, 48-108 h at 250 rpm, 108-120 h at 200 rpm) with medium volume of 100 mL added to 250 mL Erlenmeyer flasks at 30°C for 120 h cultivation. Compared to constant one-stage cultivation (medium volume of 100 mL, rotating rate of 250 rpm), the pigments were reduced by 40.4%, but citrinin was reduced by 64.2%. The most appropriate condition for the fermentation process in a 10 L fermentor is also a three-stage aeration process (0-48 h at 300 L/h, 48-96 h at 500 L/h, 96-120 h at 200 L/h) with agitation of 300 rpm at 30°C for 120 h cultivation, and 237.3 ± 5.7 U/mL pigments were produced in 120 h with 6.05 ± 0.19 mg/L citrinin in a 10 L fermentor. Compared to aeration-constant (500 L/h) cultivation, pigment production was increased by 29.6% and citrinin concentration was reduced by 79.5%. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Production and structural elucidation of exopolysaccharide from endophytic Pestalotiopsis sp. BC55.

PubMed

Mahapatra, Subhadip; Banerjee, Debdulal

2016-01-01

There is a little information on exopolysaccharide production by endophytic fungi. In this investigation endophytic Pestalotiopsis sp. BC55 was used for optimization of exopolysaccharide production. One variable at a time method and response surface methodology were adopted to find out the best culture conditions and medium compositions for maximum exopolysaccharide production. The organism produced maximum exopolysaccharide (4.320 ± 0.022 g/l EPS) in 250 ml Erlenmeyer flask containing 75 ml potato dextrose broth supplemented with (g%/l) glucose, 7.66; urea, 0.29; CaCl2, 0.05 with medium pH 6.93; after 3.76 days of incubation at 24°C. Exopolysaccharide [EPS (EP-I)] produced by this organism have Mw ∼2×10(5)Da with a melting point range of 122-124°C. Structural elucidation of the EPS (PS-I) was carried out after a series of experiments. Result indicated the presence of only (1→3)-linked β-d-glucopyranosyl moiety. The structure of the repeating unit was established as - →3)-β-d-Glcp-(1→. Copyright © 2015 Elsevier B.V. All rights reserved.

Involvement of Physical Parameters in Medium Improvement for Tannase Production by Aspergillus niger FETL FT3 in Submerged Fermentation

PubMed Central

Darah, I.; Sumathi, G.; Jain, K.; Hong, Lim Sheh

2011-01-01

Aspergillus niger FETL FT3, a local extracellular tannase producer strain that was isolated from one of dumping sites of tannin-rich barks of Rhizophora apiculata in Perak, Malaysia. This fungus was cultivated in 250 mL Erlenmeyer flask under submerged fermentation system. Various physical parameters were studied in order to maximize the tannase production. Maximal yield of tannase production, that is, 2.81 U per mL was obtained on the fourth day of cultivation when the submerged fermentation was carried out using liquid Czapek-Dox medium containing (percent; weight per volume) 0.25% NaNO3, 0.1% KH2PO4, 0.05% MgSO4 ·7H2O, 0.05% KCl, and 1.0% tannic acid. The physical parameters used initial medium pH of 6.0, incubation temperature of 30°C, agitation speed of 200 rpm and inoculums size of 6 × 106 spores/ ml. This research has showed that physical parameters were influenced the tannase production by the fungus with 156.4 percent increment. PMID:21826273

Involvement of Physical Parameters in Medium Improvement for Tannase Production by Aspergillus niger FETL FT3 in Submerged Fermentation.

PubMed

Darah, I; Sumathi, G; Jain, K; Hong, Lim Sheh

2011-01-01

Aspergillus niger FETL FT3, a local extracellular tannase producer strain that was isolated from one of dumping sites of tannin-rich barks of Rhizophora apiculata in Perak, Malaysia. This fungus was cultivated in 250 mL Erlenmeyer flask under submerged fermentation system. Various physical parameters were studied in order to maximize the tannase production. Maximal yield of tannase production, that is, 2.81 U per mL was obtained on the fourth day of cultivation when the submerged fermentation was carried out using liquid Czapek-Dox medium containing (percent; weight per volume) 0.25% NaNO(3), 0.1% KH(2)PO(4), 0.05% MgSO(4) ·7H(2)O, 0.05% KCl, and 1.0% tannic acid. The physical parameters used initial medium pH of 6.0, incubation temperature of 30°C, agitation speed of 200 rpm and inoculums size of 6 × 10(6) spores/ ml. This research has showed that physical parameters were influenced the tannase production by the fungus with 156.4 percent increment.

Regulation of the Inflammasome, a Modulator of Caspase-Mediated Cytokine Production

DTIC Science & Technology

2008-07-01

and amplified. For protein expression, insect cells were cultured in suspension at 27C using Erlenmeyer culture flasks or Cytostir bioreactors ...in bacterial or insect cell expression systems, exhibiting poor expression levels and solubility. The NALP pyrin domain has been successfully produced... mammalian codon supplementation. Protein overexpression was induced by 0.5 mM IPTG (isopropyl β-D-1- thiogalactopyranoside) in standard LB (Luria-Bertani

Application of statistical experimental methodology to optimize bioremediation of n-alkanes in aquatic environment.

PubMed

Zahed, Mohammad Ali; Aziz, Hamidi Abdul; Mohajeri, Leila; Mohajeri, Soraya; Kutty, Shamsul Rahman Mohamed; Isa, Mohamed Hasnain

2010-12-15

Response surface methodology (RSM) was employed to optimize nitrogen and phosphorus concentrations for removal of n-alkanes from crude oil contaminated seawater samples in batch reactors. Erlenmeyer flasks were used as bioreactors; each containing 250 mL dispersed crude oil contaminated seawater, indigenous acclimatized microorganism and different amounts of nitrogen and phosphorus based on central composite design (CCD). Samples were extracted and analyzed according to US-EPA protocols using a gas chromatograph. During 28 days of bioremediation, a maximum of 95% total aliphatic hydrocarbons removal was observed. The obtained Model F-value of 267.73 and probability F<0.0001 implied the model was significant. Numerical condition optimization via a quadratic model, predicted 98% n-alkanes removal for a 20-day laboratory bioremediation trial using nitrogen and phosphorus concentrations of 13.62 and 1.39 mg/L, respectively. In actual experiments, 95% removal was observed under these conditions. Copyright © 2010 Elsevier B.V. All rights reserved.

Survival of Microbial Pathogens in the Marine Environment.

DTIC Science & Technology

1978-05-01

serial 10-fold dilutions were made in ocean water sterilized by filtration (0.22 iiM Millipore membrane). The I... water (salinity 28 parts per thousand, pH 7.6) was seeded with H-i virus to contain approximately 1O4 TCID~0 1 s per ml. Four’ replicate flasks were...100 ml of untreated ocean water (test flask) and to a flask of 100 ml of filtered (0.22 pM ?4illipore membrane) ocean water (control flask). The flasks

Detecting volatile compounds from Kraft lignin degradation in the headspace of microbial cultures by selected ion flow tube mass spectrometry (SIFT-MS).

PubMed

Gibson, Andrew; Malek, Lada; Dekker, Robert F H; Ross, Brian

2015-05-01

Selected Ion Flow Tube Mass Spectrometry (SIFT-MS) was used to quantify methanol and other volatile compounds in the headspace of one bacterial and 12 fungal lignin-degrading microbial cultures. Cultures were grown in 250 mL Erlenmeyer flasks capped with aluminum foil containing 40 mL of nutrient media using Kraft lignin (0.3% w/v) as the sole carbon source. Analysis was done using SIFT-MS with H3O(+) and NO(+) precursors. Product ions were identified with multiple ion mode (MIM). Full scan (FS) mode was used to identify other compounds of interest. Absidia cylindrospora, Ischnoderma resinosum and Pholiota aurivella increased headspace methanol concentration by 136 ppb, 1196 ppb and 278 ppb, respectively, while Flammulina velutipes and Laetiporus sulphureus decreased concentration below ambient levels. F. velutipes and L. sulphureus were found to produce products of methanol oxidation (formaldehyde and formic acid) and were likely metabolizing methanol. Some additional unidentified compounds generated by the fungal cultures are intriguing and will require further study. SIFT-MS can be used to quantify methanol and other volatile compounds in the headspace of microbial cultures and has the potential to be a rapid, sensitive, non-invasive tool useful in elucidating the mechanisms of lignin degradative pathways. Copyright © 2015 Elsevier B.V. All rights reserved.

Ethanol production from agricultural wastes using Saccharomyces cerevisiae.

PubMed

Irfan, Muhammad; Nadeem, Muhammad; Syed, Quratualain

2014-01-01

The main objective of this study was production of ethanol from three lignocellulosic biomasses like sugarcane bagasse, rice straw and wheat straw by Sacchromyces cervisae. All the three substrates were ground to powder form (2 mm) and pretreated with 3%H2O2 + 2% NaOH followed by steaming at 130 °C for 60 min. These substrates were hydrolyzed by commercial cellulase enzyme. The whole fermentation process was carried out in 500 mL Erlenmeyer flask under anaerobic conditions in submerged fermentation at 30 °C for three days of incubation period. FTIR analysis of the substrates indicated significant changes in the alteration of the structure occurred after pretreatment which leads to efficient saccharification. After pretreatment the substrates were hydrolyzed by commercial cellulase enzyme and maximum hydrolysis was observed in sugarcane bagasse (64%) followed by rice straw (40%) and wheat straw (34%). Among all these tested substrates, sugarcane bagasse (77 g/L) produced more ethanol as compared to rice straw (62 g/L) and wheat straw (44 g/L) using medium composition of (%) 0.25 (NH4)2SO4, 0.1 KH2PO4, 0.05 MgSO4, 0.25 Yeast extract by S. cervisae.

Ethanol production from agricultural wastes using Sacchromyces cervisae

PubMed Central

Irfan, Muhammad; Nadeem, Muhammad; Syed, Quratualain

2014-01-01

The main objective of this study was production of ethanol from three lignocellulosic biomasses like sugarcane bagasse, rice straw and wheat straw by Sacchromyces cervisae. All the three substrates were ground to powder form (2 mm) and pretreated with 3%H2O2 + 2% NaOH followed by steaming at 130 °C for 60 min. These substrates were hydrolyzed by commercial cellulase enzyme. The whole fermentation process was carried out in 500 mL Erlenmeyer flask under anaerobic conditions in submerged fermentation at 30 °C for three days of incubation period. FTIR analysis of the substrates indicated significant changes in the alteration of the structure occurred after pretreatment which leads to efficient saccharification. After pretreatment the substrates were hydrolyzed by commercial cellulase enzyme and maximum hydrolysis was observed in sugarcane bagasse (64%) followed by rice straw (40%) and wheat straw (34%). Among all these tested substrates, sugarcane bagasse (77 g/L) produced more ethanol as compared to rice straw (62 g/L) and wheat straw (44 g/L) using medium composition of (%) 0.25 (NH4)2SO4, 0.1 KH2PO4, 0.05 MgSO4, 0.25 Yeast extract by S. cervisae. PMID:25242928

Enhanced Biotransformation of Fluoranthene by Intertidally Derived Cunninghamella elegans under Biofilm-Based and Niche-Mimicking Conditions

PubMed Central

Mitra, Sayani; Pramanik, Arnab; Banerjee, Srijoni; Haldar, Saubhik; Gachhui, Ratan

2013-01-01

The aims of the investigation were to ascertain if surface attachment of Cunninghamella elegans and niche intertidal conditions provided in a bioreactor influenced biotransformation of fluoranthene by C. elegans. A newly designed polymethylmethacrylate (PMMA) conico-cylindrical flask (CCF) holding eight equidistantly spaced rectangular strips mounted radially on a circular disc allowed comparison of fluoranthene biotransformation between CCFs with a hydrophobic surface (PMMA-CCF) and a hydrophilic glass surface (GS-CCF) and a 500-ml Erlenmeyer flask (EF). Fluoranthene biotransformation was higher by 22-fold, biofilm growth was higher by 3-fold, and cytochrome P450 gene expression was higher by 2.1-fold when C. elegans was cultivated with 2% inoculum as biofilm culture in PMMA-CCF compared to planktonic culture in EF. Biotransformation was enhanced by 7-fold with 10% inoculum. The temporal pattern of biofilm progression based on three-channel fluorescence detection by confocal laser scanning microscopy demonstrated well-developed, stable biofilm with greater colocalization of fluoranthene within extracellular polymeric substances and filaments of the biofilm grown on PMMA in contrast to a glass surface. A bioreactor with discs rotating at 2 revolutions per day affording 6-hourly emersion and immersion mimicked the niche intertidal habitat of C. elegans and supported biofilm formation and transformation of fluoranthene. The amount of transformed metabolite was 3.5-fold, biofilm growth was 3-fold, and cytochrome P450 gene expression was 1.9-fold higher in the process mimicking the intertidal conditions than in a submerged process without disc rotation. In the CCF and reactor, where biofilm formation was comparatively greater, higher concentration of exopolysaccharides allowed increased mobilization of fluoranthene within the biofilm with consequential higher gene expression leading to enhanced volumetric productivity. PMID:24038685

Development of a circulation direct sampling and monitoring system for O2 and CO2 concentrations in the gas-liquid phases of shake-flask systems during microbial cell culture.

PubMed

Takahashi, Masato; Sawada, Yoshisuke; Aoyagi, Hideki

2017-08-23

Monitoring the environmental factors during shake-flask culture of microorganisms can help to optimise the initial steps of bioprocess development. Herein, we developed a circulation direct monitoring and sampling system (CDMSS) that can monitor the behaviour of CO 2 and O 2 in the gas-liquid phases and obtain a sample without interrupting the shaking of the culture in Erlenmeyer flasks capped with breathable culture plugs. Shake-flask culturing of Escherichia coli using this set-up indicated that a high concentration of CO 2 accumulated not only in the headspace (maximum ~100 mg/L) but also in the culture broth (maximum ~85 mg/L) during the logarithmic phase (4.5-9.0 h). By packing a CO 2 absorbent in the gas circulation unit of CDMSS, a specialised shake-flask culture was developed to remove CO 2 from the headspace. It was posited that removing CO 2 from the headspace would suppress increases in the dissolved CO 2 concentration in the culture broth (maximum ~15 mg/L). Furthermore, the logarithmic growth phase (4.5-12.0 h) was extended, the U.O.D. 580 and pH value increased, and acetic acid concentration was reduced, compared with the control. To our knowledge, this is the first report of a method aimed at improving the growth of E. coli cells without changing the composition of the medium, temperature, and shaking conditions.

Identification of a New Marine Bacterial Strain SD8 and Optimization of Its Culture Conditions for Producing Alkaline Protease

PubMed Central

Cui, Hongxia; Yang, Muyang; Wang, Liping; Xian, Cory J.

2015-01-01

While much attention has been given to marine microorganisms for production of enzymes, which in general are relatively more stable and active compared to those from plants and animals, studies on alkaline protease production from marine microorganisms have been very limited. In the present study, the alkaline protease producing marine bacterial strain SD8 isolated from sea muds in the Geziwo Qinhuangdao sea area of China was characterized and its optimal culture conditions were investigated. Strain SD8 was initially classified to belong to genus Pseudomonas by morphological, physiological and biochemical characterizations, and then through 16S rDNA sequence it was identified to be likely Pseudomonas hibiscicola. In addition, the culture mediums, carbon sources and culture conditions of strain SD8 were optimized for maximum production of alkaline protease. Optimum enzyme production (236U/mL when cultured bacteria being at 0.75 mg dry weight/mL fermentation broth) was obtained when the isolate at a 3% inoculum size was grown in LB medium at 20 mL medium/100mL Erlenmeyer flask for 48h culture at 30°C with an initial of pH 7.5. This was the first report of strain Pseudomonas hibiscicola secreting alkaline protease, and the data for its optimal cultural conditions for alkaline protease production has laid a foundation for future exploration for the potential use of SD8 strain for alkaline protease production. PMID:26716833

Dredging Operations Technical Support Program. Transformation, Fixation, and Mobilization of Arsenic and Antimony in Contaminated Sediments.

DTIC Science & Technology

1984-01-01

transferred -. to a 2.8-f wide mouth erlenmeyer flask. Sufficient distiLled deionized 1 38 -... 3 .° - . N,. 0 a V0 1- f I. S.0 cc SL ’S. 0 *1’*, -E C) C 5...water-to- dry -sediment weight ratio of 15:1. The sediment-water mixtures were kept in suspension by continuous stirring with a magnetic stirrer. Sediment...pH 5.0, pit 6.5, and pH 8.0). Each Eh-pH combination was amended with 75 Wg of As(V)/g dry weight of sediment as sodium arsenate and 25 wg Sb/g dry

Pyle metaphyseal dysplasia in an African child: Case report and review of the literature.

PubMed

Wonkam, A; Makubalo, N; Roberts, T; Chetty, M

2016-05-25

Pyle disease (OMIM 265900), also known as metaphyseal dysplasia, is a rare autosomal recessive disorder with no known gene mutation. We report a case of Pyle disease in a 7-year-old African boy of mixed ancestry who presented with finger and wrist fractures following minor trauma. The radiological findings revealed abnormally broad metaphyses of the tubular bones, known as Erlenmeyer-flask bone deformity, and mild cranial sclerosis, both hallmarks of the condition. We report the first case in a patient with African ancestry, which could help in the gene discovery of this rare autosomal recessive skeletal dysplasia with unknown mutations.

27 CFR 21.94 - Acetaldol.

Code of Federal Regulations, 2014 CFR

2014-04-01

... 5 ml of this solution to a 250 ml glass-stoppered flask containing 25 ml distilled water. Add 25 ml of a freshly prepared 1 percent sodium bisulfite solution. Prepare a blank omitting the acetaldol solution. Place the flasks in a dark place away from excessive heat or cold and allow to stand six hours...

27 CFR 21.94 - Acetaldol.

Code of Federal Regulations, 2013 CFR

2013-04-01

... 5 ml of this solution to a 250 ml glass-stoppered flask containing 25 ml distilled water. Add 25 ml of a freshly prepared 1 percent sodium bisulfite solution. Prepare a blank omitting the acetaldol solution. Place the flasks in a dark place away from excessive heat or cold and allow to stand six hours...

27 CFR 21.94 - Acetaldol.

Code of Federal Regulations, 2011 CFR

2011-04-01

... 5 ml of this solution to a 250 ml glass-stoppered flask containing 25 ml distilled water. Add 25 ml of a freshly prepared 1 percent sodium bisulfite solution. Prepare a blank omitting the acetaldol solution. Place the flasks in a dark place away from excessive heat or cold and allow to stand six hours...

27 CFR 21.94 - Acetaldol.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 5 ml of this solution to a 250 ml glass-stoppered flask containing 25 ml distilled water. Add 25 ml of a freshly prepared 1 percent sodium bisulfite solution. Prepare a blank omitting the acetaldol solution. Place the flasks in a dark place away from excessive heat or cold and allow to stand six hours...

27 CFR 21.94 - Acetaldol.

Code of Federal Regulations, 2012 CFR

2012-04-01

... 5 ml of this solution to a 250 ml glass-stoppered flask containing 25 ml distilled water. Add 25 ml of a freshly prepared 1 percent sodium bisulfite solution. Prepare a blank omitting the acetaldol solution. Place the flasks in a dark place away from excessive heat or cold and allow to stand six hours...

[Studies for analyzing the prohibited ingredients such as acetohexamide in cosmetics].

PubMed

Tokunaga, Hiroshi; Uchino, Tadashi

2005-01-01

Acetohexamide (AH) is nominated as the prohibited ingredients in cosmetics in Japanese Pharmaceutical Affairs Act. So the analytical method for AH was investigated by HPLC. The lotion or milky lotion of 0.5g was put into a 10-ml volumetric flask. After adding 1.0ml of AH solution at 50 microg/ml into the volumetric flask, the mixture was made up to 10ml with methanol as the testing solution. Creams were procedured as follows; 0.5 g of cream was put into a 10-ml volumetric flask. After adding 1.0ml of tetrahydrofuran into the volumetric flask, the mixture was stirred for several minutes and the ingredients of the creams were dissolved. After adding 1.0ml of AH solution at 50 microg/ml into the volumetric flask, the mixture was made up to 10ml with methanol. One milliliter of the mixture including AH at 5 microg/ml was exactly put into a test tube with a cap and then 1 ml of water and 1 ml of hexane were added. After shaking vigorously, stand for several minutes. After centrifuging, the hexane layer was eliminated and the residual mixture was used as the test solution. The testing solution of 20 microl was analyzed by HPLC using the ODS column (CAPCELL PAK C18 column, 4.6 x 250mm), the mixture of acetonitrile and 50 mmol/l phosphate buffer(pH 5.3)(3:1) and the detection wavelength of 247 nm. The working curve from 0.5 to 6.0 microg/ml showed a linear line between the concentrations of AH and the peak areas. There was no interference of peak of AH with the ingredients such as methylparaben, ethylparaben in the lotions, milky lotion and creams.

[Studies for analyzing the prohibited ingredients such as cyproheptadine hydrochloride in cosmetics].

PubMed

Tokunaga, Hiroshi; Uchino, Tadashi

2005-01-01

Cyproheptadine hydrochloride (CH) is nominated as the prohibited ingredients in cosmetics in Japanese Pharmaceutical Affairs Act. So the analytical method for CH was investigated by HPLC. The lotion or milky lotion of 0.5 g was put into a 10-ml volumetric flask. After adding 1.0ml of CH solution at 50 microg/ml into the volumetric flask, the mixture was made up to 10ml with methanol as the test solution. Creams were procedured as follows; 0.5 g of cream was put into a 10-ml volumetric flask. After adding 1.0 ml of tetrahydrofuran into the volumetric flask, the mixture was stirred for several minutes and the ingredients of the creams were dissolved. After adding 1.0 ml of CH solution at 50 microg/ml into the volumetric flask, the mixture was made up to 10ml with methanol. This mixture was transferred to a centrifuging tube with a cap and then the tube was centrifuged for 5 minutes at 3000 rpm. The supernatant was used as the test solution. The test solution of 20 microl was analyzed by HPLC using the ODS column (CAPCELL PAK C18 column, 4.6 x 250 mm), the mixture of 1% acetic acid with 10 mmol/l sodium octanesulfonate and acetonitrile (11:9) and the detection wavelength of 286 nm. The working curve from 0.5 to 6.0 microg/ml showed a linear line between the concentrations of CH and the peak areas. There was no interference of peak of CH with the ingredients such as methylparaben, ethylparaben in the lotions, milky lotion and creams.

[Studies for analyzing restricted ingredients such as phenylbenzoimidazole sulfonic acid].

PubMed

Tokunaga, Hiroshi; Mori, Kenichiro; Onuki, Nahomi; Nosaka, Tomio; Doi, Kayo; Sakaguchi, Hiroshi; Fujii, Makiko; Takano, Katuhiro; Hayashi, Masato; Yoshizawa, Kenichi; Shimamura, Kimio; Sato, Nobuo

2006-01-01

Phenylbenzoimidazol sulfonic acid (PBS) is a kind of sunscreens in cosmetics and is nominated as the restricted ingredients in cosmetics in Japanese Pharmaceutical Affairs Act. So the analytical method for PBS was investigated by HPLC. 1.0 g of the lotions with 1.0% PBS was exactly weighed, put into a 50-mL volumetric flask. Water was added to make exactly 50 mL and this mixture was used as the sample solution. On the other hand, 1.0 g of the creams with 1.0% PBS was exactly weighed, put into a beaker. After adding 1 mL of tetrahydrofuran and dissolving the cream, that mixture was transferred to a 50-mL volumetric flask. And then the beaker was rinsed with 1 mL of tetrahydrofuran and the rinsed solution was put together into the volumetric flask. After adding water to the volumetric flask to make exactly 50 mL, this mixture was used as the sample solution. If necessary, the mixture was filtrated with a membrane filter (0.45 microm). 5.0 mL of the sample solution was pipetted and put into a 200-mL volumetric flask. After adding water to make exactly 200 mL, 20 microL of this solution was analyzed by HPLC using the ODS column (CAPCELL PAK C18 column, 4.6 mm i.d. x 250 mm), the mixture of 40 mmol/L acetic buffer (pH 3.4) and acetonitrile (3:1) with 0.8 mmol/L dodecyltrimethyl ammonium bromide and the detection wavelength of 305 nm. The working curve from 0.5 to 20.0 microg/mL showed a linear line between the concentrations of PBS and the peak areas. There was no interference of peak of PBS from the lotion and cream.

Comparison of a 50 mL pycnometer and a 500 mL flask, EURAMET.M.FF.S8 (EURAMET 1297)

NASA Astrophysics Data System (ADS)

Mićić, Ljiljana; Batista, Elsa

2018-01-01

The purpose of this comparison was to compare the results of the participating laboratories in the calibration of 50 mL pycnometer and 500 mL volumetric flask using the gravimetric method. Laboratories were asked to determined the 'contained' volume of the 50 mL pycnometer and of the 500 mL flask at a reference temperature of 20 °C. The gravimetric method was used for both instruments by all laboratories. Main text To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

Modifications to the algal growth inhibition test for use as a regulatory assay

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

Geis, S.W.; Fleming, K.L.; Korthals, E.T.

2000-01-01

Biological assays using aquatic invertebrates and fish do not necessarily predict protection levels for primary producers such as algae and aquatic macrophytes. State regulatory programs may not be protecting the environment from many phytotoxic compounds. Recent modifications of the US Environmental Protection Agency's algal test were evaluated for their potential use as a regulatory assay. Primary goals of this investigation were to downsize the algal assay and to evaluate various methods of automation. Disposable microplates with 2-ml sample wells were evaluated as an alternative testing chamber for the 96-h growth inhibition test with Raphidocelis subcapitata (formerly known as Selenastrum capricornutum).more » The authors compared the standardized Erlenmeyer {reg_sign} flask test to the microplate test using CuCl{sub 2}, NaCl, phenol, ZnCl{sub 2}, and a surfactant. They noted improved control performance with the microplate test, whereas median inhibitory concentration values were similar for both methods. Other procedures they addressed included the use of EDTA, filtration of samples, and the effect of colored samples on algal growth. They also evaluated growth estimates by comparing manual cell counting to more automated growth estimates using fluorescence and absorbance endpoints. The use of fluorescence and absorbance measurements demonstrated reductions in replicate variability over manual counting and may offer time-saving alternatives for laboratory analysts.« less

[Optimization of fermentation conditions for cold-adapted amylase production by Micrococcus antarcticus and its enzymatic properties].

PubMed

Fan, Hong-xi; Liu, Ying; Liu, Zhi-pei

2009-08-15

By single factor experiments, the fermentation conditions for cold-adapted amylase production from Micrococcus antarcticus were determined as follows(medium g/L): Na2 HPO4 2.0, KH2PO4 1.0, MgSO4 x 7H2O 0.1, NaCl 5.0, (NH4)2SO4 2.5, maltose 5.0, trace element solution 5.0 mL, pH 8.0, 100 mL/Erlenmeyer flask (500 mL); cultivation was in a rotating shaker at 12 degrees C and 160 r/min for 64 h.Under those conditions,the highest total enzyme activity (2.6 U/mL) was obtained and increased by 10.8 fold compared with the original value of 0.24 U/mL before optimization. This amylase was purified by concentration with ultrafiltration membrane module, Hitrap Q anion exchange chromatography and Superdex 200 gel filtration chromatography. The optimal temperature and pH for the purified amylase were 30 degrees C and 6.0, respectively.It still showed high activity at low temperature 10-15 degrees C. It was sensitive to high temperature but was stable at pH 6.0-10.0 with at least 70% activity remained. These results indicated that it was a typical cold-adapted enzyme. The enzyme activity was stimulated by Ca2+, Mn2+, Co2+ and Mg2+; but inhibited by Zn2+, Ba2+, Ag+, Cu2+, Al3+, Fe2, Fe3+, Hg2+, EDTA and citrate. This cold-adapted amylase showed resistance to inactivation of 0.1% nonionic surfactants such as Tween 80, TrintonX-100, etc. Its Km was 0.90 mg/mL.

Laboratory and Field Evaluation of a Waterless Food Service Sanitation System Used by Military Mobile Kitchen Trailer Crews

DTIC Science & Technology

1993-12-01

Stopper the flask and shake vigorously. The mixture was titrated with 0.003 N sodium lauryl sulfate drcpwise. The endpoint was the first definite...100 g sodium sulfate and 1000 mL distilled water, pill0) and three drops of 0.1% bracu 1henol blue indicator to 50 ML of sample in a 250 mL flask...Product QDS (Syncide Plus) (1) was determined by a bromcphenol blue nethod (7). Add 25 mL of chloroform, 25 mL salt buffer solution (7 g sodium carbonate

Simplified Method of the Growth of Human Tumor Infiltrating Lymphocytes (TIL) in Gas-Permeable Flasks to Numbers Needed for Patient Treatment

PubMed Central

Jin, Jianjian; Sabatino, Marianna; Somerville, Robert; Wilson, John R.; Dudley, Mark E.; Stroncek, David F.; Rosenberg, Steven A.

2012-01-01

Adoptive cell therapy (ACT) of metastatic melanoma with autologous tumor infiltrating lymphocytes (TIL) is clinically effective, but TIL production can be challenging. Here we describe a simplified method for initial TIL culture and rapid expansion in gas-permeable flasks. TIL were initially cultured from tumor digests and fragments in 40 mL capacity flasks with a 10 cm2 gas-permeable silicone bottom, G-Rex10. A TIL rapid expansion protocol (REP) was developed using 500 mL capacity flasks with a 100 cm2 gas-permeable silicone bottom, G-Rex100. TIL growth was successfully initiated in G-Rex10 flasks from tumor digests from 13 of 14 patients and from tumor fragments in all 11 tumor samples tested. TIL could then be expanded to 8–10×109 cells in a two-step REP which began by seeding 5 × 106 TIL into a G-Rex100 flask, followed by expansion at day 7 into 3 G-Rex100 flasks. To obtain the 30 to 60 × 109 cells used for patient treatment we seeded 6 G-Rex100 flasks with 5×106 cells and expanded into 18 G-Rex100 flasks. Large scale TIL REP in gas-permeable flasks requires approximately 9 to 10 liters of media, about 3 to 4 times less than other methods. In conclusion, TIL initiation and REP in gas-permeable G-Rex flasks require fewer total vessels, less media, less incubator space and less labor than initiation and REP in 24-well plates, tissue culture flasks and bags. TIL culture in G-Rex flasks will facilitate the production of TIL at the numbers required for patient treatment at most cell processing laboratories. PMID:22421946

Aerosol-phase Activity of Iodine Captured from a Triiodide Resin Filter on Fine Particles Containing an Infectious Virus

DTIC Science & Technology

2015-01-01

conductive tubing direc ted the aerosol into a heat sealed plastic bag (Food Saver; Jarden Consumer Solutions, Rye, NY), measuring 589 cm 9 254...a self sealing injection port attached to each bag with an adhesive. Approximately 100 mg of iodine crystals were stored at 35°C in an Erlenmeyer...was collected in a bag, 075 ml was drawn from the headspace into a 1 ml syringe and needle and immediately injected through the port into the sample

An efficient method to control high mannose and core fucose levels in glycosylated antibody production using deoxymannojirimycin.

PubMed

Shalel Levanon, Sagit; Aharonovitz, Orit; Maor-Shoshani, Ayelet; Abraham, Gita; Kenett, Dan; Aloni, Yehoshua

2018-06-20

Glycosylation on the Fc region of recombinant Immunoglobulin G (IgG) therapeutic antibodies is a critical protein quality attribute which may affect the efficacy and safety of the molecule. During the development of biosimilar therapeutics, adjustment of the glycosylation profile is required in order to match the reference innovator profile. Deoxymannojirimycin (DMJ), a known inhibitor of mannosidase, was used in this study to modulate the glycosylation pattern of antibodies. The effect of DMJ, at concentrations of 5 μM - 500 μM, on non-fucosylated glycoform levels was tested in the biosynthesis processes of two different IgG1 (IgG1 #A and IgG1 #B) using two Chinese hamster ovary (CHO) cell lines (CHO-DXB-11 and CHOK1SV, respectively) in Erlenmeyer flasks and in lab scale bioreactors. DMJ affected glycan forms in a dose response manner. At the highest concentration tested, DMJ reduced N-linked complex glycoform and core fucose levels by 15 and 14 fold, respectively, and increased high mannose level by 21 fold. 10 μM DMJ decreased IgG1 #A core fucose level in CHO-DXB-11 from 92% to 73% and increased high mannose level from 4% to 22% in Erlenmeyer flasks. Furthermore, in lab scale bioreactors, 15 μM DMJ decreased IgG1 #A core fucose level from 95% to 84% and increased high mannose level from 3% to 13%. Core fucose level of IgG1 #B in CHOK1SV was decreased from 81% to 73% using 10 μM DMJ in lab scale bioreactors while high mannose was increased from 6% to 15%. While affecting core fucose and high mannose levels, DMJ decreased maximum viable cell concentration by 16% and did not significantly affect cell productivity (less than 10%). This study demonstrated that DMJ can enable the control of core fucosylated and high mannose levels of IgG1 antibodies in a defined range. Copyright © 2018 Elsevier B.V. All rights reserved.

Using Ice-Cooled Condensers in Chemistry Laboratory

NASA Astrophysics Data System (ADS)

Solomon, Sally; Brook, Bryan; Rutkowsky, Susan; Bennet, Joseph

2003-03-01

An ice-cooled condenser, consisting of a jacket built around a tube open to the atmosphere with an outlet for removal of melting ice, is designed for use in academic laboratory classes. The apparatus can be used in place of standard water cooled condensers in setups where refluxing or distillation is performed. With this simple, inexpensive device there is no need for access to running water. Potential flooding due to insecure tubing is no longer a problem. The ice-cooled accessory, produced with standard glass tubing and either 14/10 or 14/20 ground glass joints, is compatible with most commercially available microscale or small scale kits. The device may even be used with an Erlenmeyer flask and a stopper or cork. Two experiments using ordinary household chemicals are suggested, one requiring refluxing and the other distillation.

Production, purification and characterization of an aspartic protease from Aspergillus foetidus.

PubMed

Souza, Paula Monteiro; Werneck, Gabriela; Aliakbarian, Bahar; Siqueira, Felix; Ferreira Filho, Edivaldo Ximenes; Perego, Patrizia; Converti, Attilio; Magalhães, Pérola Oliveira; Junior, Adalberto Pessoa

2017-11-01

An acidic thermostable protease was extracellularly produced either in shake flask or in stirred tank bioreactor by an Aspergillus foetidus strain isolated from the Brazilian savanna soil using different nitrogen sources. Its maximum activity (63.7 U mL -1 ) was obtained in a medium containing 2% (w/v) peptone. A cultivation carried out in a 5.0 L stirred-tank bioreactor provided a maximum protease activity 9% lower than that observed in Erlenmeyer flasks, which was obtained after a significantly shorter (by 16-29%) time. Protease purification by a combination of gel-filtration chromatography resulted in a 16.9-fold increase in specific activity (248.1 U g -1 ). The estimated molecular weight of the purified enzyme was 50.6 kDa, and the optimal pH and temperature were 5.0 and 55 °C, respectively. The enzyme was completely inhibited by pepstatin A, and its activity enhanced by some metals. According to the inhibition profiles, it was confirmed that the purified acid protease belongs to the aspartic protease type. These results are quite promising for future development of large-scale production of such protease, which can be useful in biotechnological applications requiring high enzyme activity and stability under acidic conditions. Copyright © 2017 Elsevier Ltd. All rights reserved.

Cashew apple juice as microbial cultivation medium for non-immunogenic hyaluronic acid production.

PubMed

Oliveira, Adriano H; Ogrodowski, Cristiane C; de Macedo, André C; Santana, Maria Helena A; Gonçalves, Luciana R B

2013-12-01

In this work, natural cashew apple juice was used as cultivation medium as an alternative to substitute brain heart infusion medium. The effect of aeration and juice supplementation with yeast extract on the production of hyaluronic acid in batch fermentation was also investigated. Similar levels of cell mass were obtained in inoculum using cashew apple juice supplemented with yeast extract or the conventional brain heart infusion medium. Fermentation in Erlenmeyer flasks produced low biomass and hyaluronic acid concentrations. The hyaluronic acid concentration and viscosity increased from 0.15 g/L and 3.87 cP (no aeration or medium supplementation) to 1.76 g/L and 107 cP, when aeration (2 vvm) and 60 g/L of yeast extract were used. The results suggest the production of low-molecular weight hyaluronic acid oligomers instead of the high molecular weight polymer.

Cashew apple juice as microbial cultivation medium for non-immunogenic hyaluronic acid production

PubMed Central

Oliveira, Adriano H.; Ogrodowski, Cristiane C.; de Macedo, André C.; Santana, Maria Helena A.; Gonçalves, Luciana R.B.

2013-01-01

In this work, natural cashew apple juice was used as cultivation medium as an alternative to substitute brain heart infusion medium. The effect of aeration and juice supplementation with yeast extract on the production of hyaluronic acid in batch fermentation was also investigated. Similar levels of cell mass were obtained in inoculum using cashew apple juice supplemented with yeast extract or the conventional brain heart infusion medium. Fermentation in Erlenmeyer flasks produced low biomass and hyaluronic acid concentrations. The hyaluronic acid concentration and viscosity increased from 0.15 g/L and 3.87 cP (no aeration or medium supplementation) to 1.76 g/L and 107 cP, when aeration (2 vvm) and 60 g/L of yeast extract were used. The results suggest the production of low-molecular weight hyaluronic acid oligomers instead of the high molecular weight polymer. PMID:24688498

Effect of temperature on growth and paralytic toxin profiles in isolates of Gymnodinium catenatum (Dinophyceae) from the Pacific coast of Mexico.

PubMed

Band-Schmidt, Christine J; Bustillos-Guzmán, José J; Hernández-Sandoval, Francisco E; Núñez-Vázquez, Erick J; López-Cortés, David J

2014-11-01

The effects of temperature on growth, cell toxicity, toxin content, and profile of paralytic shellfish toxins was determined in eight isolates of Gymnodinium catenatum from several localities along the Pacific Coast of Mexico. The isolates were cultivated in modified f/2 media with Se (10(-8) M), and a reduced concentration of Cu (10(-8) M), under a 12 h:12 h day-night cycle with an irradiance of 150 μE m(-2) s(-1). Isolates were progressively adapted for three generations to each of the temperatures (16, 19, 22, 24, 27, 30, and 33 °C). The cultures were grown in 125 mL Erlenmeyer flasks with 60 mL of media and harvested by filtration in late exponential growth. Toxins were analyzed by HPLC with a post-column oxidation and fluorescent detection (FLD). G. catenatum isolates tolerate temperatures between 16 and 33 °C, with maximum growth rates of 0.32 and 0.39 div day(-1) at 21 °C and 24 °C, respectively; maximum cell densities of 4700 and 5500 cells mL(-1) were obtained at 27 and 21 °C, respectively. No effect of toxicity per cell with temperature was observed, varying between 10.10 and 28.19 pgSXTeq cell(-1). Ten saxitoxin analogues were detected in all isolates, observing changes in the toxin profile with temperature. C1/2 toxins decreased from 80% mol at 16 °C to 20% mol at 33 °C, B1/2 toxins increased from 19% mol at 16 °C to 42% mol at 33 °C, and decarbamoyl toxins were more abundant at 21 °C. These results show that G. catenatum isolates from different regions of the Pacific coast of Mexico have a similar response to temperature and that this parameter can modify growth rate, cell density, and toxin profile of the species, particularly the decarbamoyl and sulfocarbamoyl toxins. Copyright © 2014 Elsevier Ltd. All rights reserved.

Sugarcane straw as a feedstock for xylitol production by Candida guilliermondii FTI 20037.

PubMed

Hernández-Pérez, Andrés Felipe; de Arruda, Priscila Vaz; Felipe, Maria das Graças de Almeida

2016-01-01

Sugarcane straw has become an available lignocellulosic biomass since the progressive introduction of the non-burning harvest in Brazil. Besides keeping this biomass in the field, it can be used as a feedstock in thermochemical or biochemical conversion processes. This makes feasible its incorporation in a biorefinery, whose economic profitability could be supported by integrated production of low-value biofuels and high-value chemicals, e.g., xylitol, which has important industrial and clinical applications. Herein, biotechnological production of xylitol is presented as a possible route for the valorization of sugarcane straw and its incorporation in a biorefinery. Nutritional supplementation of the sugarcane straw hemicellulosic hydrolyzate as a function of initial oxygen availability was studied in batch fermentation of Candida guilliermondii FTI 20037. The nutritional supplementation conditions evaluated were: no supplementation; supplementation with (NH4)2SO4, and full supplementation with (NH4)2SO4, rice bran extract and CaCl2·2H2O. Experiments were performed at pH 5.5, 30°C, 200rpm, for 48h in 125mL Erlenmeyer flasks containing either 25 or 50mL of medium in order to vary initial oxygen availability. Without supplementation, complete consumption of glucose and partial consumption of xylose were observed. In this condition the maximum xylitol yield (0.67gg(-1)) was obtained under reduced initial oxygen availability. Nutritional supplementation increased xylose consumption and xylitol production by up to 200% and 240%, respectively. The maximum xylitol volumetric productivity (0.34gL(-1)h(-1)) was reached at full supplementation and increased initial oxygen availability. The results demonstrated a combined effect of nutritional supplementation and initial oxygen availability on xylitol production from sugarcane straw hemicellulosic hydrolyzate. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

Purification and characterization of a new neutral metalloprotease from marine Exiguobacterium sp. SWJS2.

PubMed

Lei, Fenfen; Cui, Chun; Zhao, Haifeng; Tang, Xuelu; Zhao, Mouming

2016-01-01

Among the protease-producing bacterial strains isolated from deep-sea sediments, SWJS2 was finally selected and identified as genus Exiguobacterium. Plackett-Burman and orthogonal array designs were applied to optimize the fermentation conditions, and the results are as follows: Glucose 5g, yeast extract 15g, glycerin 2g and CaCl2 ⋅2H2 O 0.5 g dissolved in 1 L artificial seawater; temperature 25 °C, original pH 7, inoculum rate 2%, seed age 12 H, loading volume 25 mL (250-mL Erlenmeyer flask), shaking speed 150 rpm, and fermentation time 44 H. The protease activity production was improved from about 80 to 660 U/mL under the optimized parameters. The protease was purified fourfold with specificity activity of 30,654.1 U/mg protein and a total yield of 16.2%. The protease exhibited the maximum activity at 40-45 °C and pH 7. Moreover, the enzyme activity was found to be inhibited by Cu(2+) , Ba(2+) , Cd(2+) , Hg(2+) , and Al(3+) at 5 mM, whereas it can be increased by Mg(2+) , Mn(2+) , and Ca(2+) at 0.5-5 mM. The enzyme was totally inactivated by 1 or 5 mM ethylenediaminetetraacetic acid but not by phenylmethanesulfonyl fluoride, tyrpsin inhibitor from Glycine max (STI), benzamidine, 5,5'-dithio-bis-(2-nitro benzoic acid), or pepstatin A, suggesting that it belonged to metalloprotease. © 2015 International Union of Biochemistry and Molecular Biology, Inc.

Biodegradation of total organic carbons (TOC) in Jordanian petroleum sludge.

PubMed

Mrayyan, Bassam; Battikhi, Mohammed N

2005-04-11

Biodegradation is cost-effective, environmentally friendly treatment for oily contaminated sites by the use of microorganisms. In this study, laboratory experiments were conducted to establish the performance of bacterial isolates in degradation of organic compounds contained in oily sludge from the Jordanian Oil Refinery plant. As a result of the laboratory screening, three natural bacterial consortia capable of degrading total organic carbons (TOC) were prepared from isolates enriched from the oil sludge. Experiments were conducted in Erlenmeyer flasks under aerobic conditions, with TOC removal percentage varied from 0.3 to 28% depending on consortia type and concentration. Consortia 7B and 13B exhibited the highest TOC removal percentage of 28 and 22%, respectively, before nutrient addition. TOC removal rate was enhanced after addition of nutrients to incubated flasks. The highest TOC reduction (43%) was estimated after addition of combination of nitrogen, phosphorus and sulphur to consortia 7B. A significant variation (P<0.005) was observed between the effect of consortia type and concentration on TOC% reduction. No significant variation was observed between incubation at 10 and 18 days in TOC% reduction. This is the first report concerning biological treatment of TOC by bacteria isolated from the oil refinery plants, where it lays the ground for full integrated studies recommended for the degradation of organic compounds that assist in solving sludge problems.

Formation, Structure and Properties of Boron Nitride Fibers from Polymer Precursors

DTIC Science & Technology

1992-02-25

conversion, have been demonstrated in this study. Poly(ethylene oxide) (PEO), poly(N-vinyl-2-pyrrolidinone) ( PVP ), and poly(methyl methacrylate) (PMMA) were...vinylpyrrolidinone ( PVP , Mw = 360,000). A 100 ml reaction flask with teflon boiling chips and a 100 ml receiving flask were used on a high boiling distilling...CH 3 OBO) 3 : B(OH)3 Predicted 2 Experiment Experiment, 1 wt% PVP 2:1 47.8 mole% CH 3 OH 39.6 mole% CH 3OH 35.2 mole% CH3 OH 3:1 47.8 - 35.5 4:1 47.8

Production of R,R-2,3-butanediol of ultra-high optical purity from Paenibacillus polymyxa ZJ-9 using homologous recombination.

PubMed

Zhang, Li; Cao, Can; Jiang, Ruifan; Xu, Hong; Xue, Feng; Huang, Weiwei; Ni, Hao; Gao, Jian

2018-08-01

The present study describes the use of metabolic engineering to achieve the production of R,R-2,3-butanediol (R,R-2,3-BD) of ultra-high optical purity (>99.99%). To this end, the diacetyl reductase (DAR) gene (dud A) of Paenibacillus polymyxa ZJ-9 was knocked out via homologous recombination between the genome and the previously constructed targeting vector pRN5101-L'C in a process based on homologous single-crossover. PCR verification confirmed the successful isolation of the dud A gene disruption mutant P. polymyxa ZJ-9-△dud A. Moreover, fermentation results indicated that the optical purity of R,R-2,3-BD increased from about 98% to over 99.99%, with a titer of 21.62 g/L in Erlenmeyer flasks. The latter was further increased to 25.88 g/L by fed-batch fermentation in a 5-L bioreactor. Copyright © 2018 Elsevier Ltd. All rights reserved.

Development of a semidefined growth medium for Pedobacter cryoconitis BG5 using statistical experimental design.

PubMed

Ong, Magdalena; Ongkudon, Clarence M; Wong, Clemente Michael Vui Ling

2016-10-02

Pedobacter cryoconitis BG5 are psychrophiles isolated from the cold environment and capable of proliferating and growing well at low temperature regime. Their cellular products have found a broad spectrum of applications, including in food, medicine, and bioremediation. Therefore, it is imperative to develop a high-cell density cultivation strategy coupled with optimized growth medium for P. cryoconitis BG5. To date, there has been no published report on the design and optimization of growth medium for P. cryoconitis, hence the objective of this research project. A preliminary screening of four commercially available media, namely tryptic soy broth, R2A, Luria Bertani broth, and nutrient broth, was conducted to formulate the basal medium. Based on the preliminary screening, tryptone, glucose, NaCl, and K2HPO4 along with three additional nutrients (yeast extract, MgSO4, and NH4Cl) were identified to form the basal medium which was further analyzed by Plackett-Burman experimental design. Central composite experimental design using response surface methodology was adopted to optimize tryptone, yeast extract, and NH4Cl concentrations in the formulated growth medium. Statistical data analysis showed a high regression factor of 0.84 with a predicted optimum optical (600 nm) cell density of 7.5 using 23.7 g/L of tryptone, 8.8 g/L of yeast extract, and 0.7 g/L of NH4Cl. The optimized medium for P. cryoconitis BG5 was tested, and the observed optical density was 7.8. The cost-effectiveness of the optimized medium was determined as 6.25 unit prices per gram of cell produced in a 250-ml Erlenmeyer flask.

Citric Acid Production from Orange Peel Wastes by Solid-State Fermentation

PubMed Central

Torrado, Ana María; Cortés, Sandra; Manuel Salgado, José; Max, Belén; Rodríguez, Noelia; Bibbins, Belinda P.; Converti, Attilio; Manuel Domínguez, José

2011-01-01

Valencia orange (Citrus sinensis) peel was employed in this work as raw material for the production of citric acid (CA) by solid-state fermentation (SSF) of Aspergillus niger CECT-2090 (ATCC 9142, NRRL 599) in Erlenmeyer flasks. To investigate the effects of the main operating variables, the inoculum concentration was varied in the range 0.5·103 to 0.7·108 spores/g dry orange peel, the bed loading from 1.0 to 4.8 g of dry orange peel (corresponding to 35-80 % of the total volume), and the moisture content between 50 and 100 % of the maximum water retention capacity (MWRC) of the material. Moreover, additional experiments were done adding methanol or water in different proportions and ways. The optimal conditions for CA production revealed to be an inoculum of 0.5·106 spores/g dry orange peel, a bed loading of 1.0 g of dry orange peel, and a humidification pattern of 70 % MWRC at the beginning of the incubation with posterior addition of 0.12 mL H2O/g dry orange peel (corresponding to 3.3 % of the MWRC) every 12 h starting from 62 h. The addition of methanol was detrimental for the CA production. Under these conditions, the SSF ensured an effective specific production of CA (193 mg CA/g dry orange peel), corresponding to yields of product on total initial and consumed sugars (glucose, fructose and sucrose) of 376 and 383 mg CA/g, respectively. These results, which demonstrate the viability of the CA production by SSF from orange peel without addition of other nutrients, could be of interest to possible, future industrial applications. PMID:24031646

Production of microsclerotia by Brazilian strains of Metarhizium spp. using submerged liquid culture fermentation.

PubMed

Mascarin, Gabriel Moura; Kobori, Nilce Naomi; de Jesus Vital, Rayan Carlos; Jackson, Mark Alan; Quintela, Eliane Dias

2014-05-01

We investigated the potential production and desiccation tolerance of microsclerotia (MS) by Brazilian strains of Metarhizium anisopliae (Ma), M. acridum (Mc) and M. robertsii (Mr). These fungi were grown in a liquid medium containing 16 g carbon l⁻¹ with a carbon:nitrogen ratio of 50:1. One hundred milliliters cultures were grown in 250 ml Erlenmeyer flasks in a rotary incubator shaker at 28 °C and 200 rpm for 5 days. Five-day-old MS were harvested, mixed with diatomaceous earth (DE) and air-dried for 2 days at 30 °C. The air-dried MS-DE granular preparations were milled by mortar + pestle and stored in centrifuged tubes at either 26 or -20 °C. Desiccation tolerance and conidia production were assessed for dried MS granules by measuring hyphal germination after incubation for 2 days on water agar plates at 26 °C and for conidia production following 7 days incubation. Yields of MS by all strains of Metarhizium were 6.1-7.3 × 10⁶ l⁻¹ after 3 days growth with maximum MS yields (0.7-1.1 × 10⁷ l⁻¹) after 5 days growth. No differences in biomass accumulation were observed after 3 days growth, whereas Ma-CG168 showed the highest biomass accumulation after 5 days growth. Dried MS-DE preparations of all fungal strains were equally tolerant to desiccation (≥93 % germination) and the highest conidia production was obtained by MS granules of Mc-CG423 (4 × 10⁹ conidia g⁻¹). All MS granules showed similar stability after storage at either 26 or -20 °C for 3.5 months.

In vitro penetration of swine oocytes by homologous spermatozoa: Distinct systems for gamete's co-incubation and oocyte's cryopreservation.

PubMed

Macedo, M C; Lucia, T; Rambo, G; Ferreira Filho, E B; Rosa, A P; Fabiane, C; Cabral, M; Deschamps, J C

2010-02-01

In vitro penetration (IVP) of swine oocytes by homologous spermatozoa was evaluated in two experiments using four boars as semen donors. In experiment 1, the IVP rate and the number of penetrating spermatozoa (PSP) were compared using three co-incubation systems for vitrified oocytes and fresh sperm: (1) 35mL petri dishes in a CO(2) incubator, (2) 35mL petri dishes in bags (submarine system) and (3) glass flasks partially submerged in water bath with the same gas mixture used for the bag system. Mean PSP was 8.2+/-10.1 and the IVP rate was 90.5%. The PSP differed across all systems (P=0.0006): 15.5+/-0.5 for flasks, 6.3+/-0.4 for CO(2), and 3.9+/-0.4 for bags. The IVP rate for flasks (95.0%) was greater (P=0.01) than for CO(2) and bags (90.8% and 85.0%, respectively), but it did not differ between flasks and CO(2) for three boars (P>0.05). In experiment 2, co-incubation was done as described for glass flasks in experiment 1. The IVP rate and PSP were compared for cryopreserved oocytes: either vitrified in open pulled straws (OPS), or frozen in cryotubes. Mean PSP was 5.4+/-6.5 and IVP rate was 89.6%. Both PSP and IVP rate were greater (P<0.0001) for oocytes frozen in cryotubes (7.0+/-0.3% and 95.8%, respectively) than those frozen in OPS (3.7+/-0.3% and 83.4%, respectively), with no differences found for three boars (P>0.05). In summary, successful IVP of swine oocytes by homologous spermatozoa can be achieved using gametes incubated in glass flasks and oocytes frozen in cryotubes.

[Studies for analyzing prohibited ingredients such as estriol in cosmetics].

PubMed

Tokunaga, Hiroshi; Takeuchi, Orie; Uchino, Tadashi; Ando, Masanori

2004-01-01

Estriol (EO) is nominated as the prohibited ingredients in cosmetics in Japanese Pharmaceutical Affairs Act. So the analytical method using HPLC for EO was investigated. After placing 1.0 ml of EO solution at 50 microg/ml and 0.5 g of the lotion into a 10-ml volumetric flask, the methanol was added to make until that volume and this solution was used as the testing solution. Milky lotion was procedured as follows: After placing 1.0 ml of EO solution at 50 microg/ml and 0.5 g of the milky lotion into a 10-ml volumetric flask, the methanol was added to make until that volume. The suspending mixture was moved to a centrifuging tube with a cap. After centrifuging at 3000 rpm for 5 minutes, the supernatant was used as the testing solution. The testing solution of 20 microl was determined by HPLC using the ODS column (CAPCELL PAK C18 column, 4.6 x 250 mm), the mixture of water and acetonitrile (31:9) and the detection wavelength of 285 nm. The working curve from 1.0 to 6.0 microg/ml showed a linear line between the concentrations of EO and the peak area. There was no interference of peak of EO from the lotion and milky lotion.

Adsorptive loss of secreted recombinant proteins in transgenic rice cell suspension cultures.

PubMed

Kwon, Jun-Young; Lee, Kyoung-Hoon; Cheon, Su-Hwan; Ryu, Hyun-Nam; Kim, Sun Jin; Kim, Dong-Il

2012-03-01

Adsorptive loss of human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) in transgenic rice cell suspension cultures was investigated using glass flasks, plastic flasks, disposable vessels, and stainless steel vessels. When hCTLA4Ig was added to the glass flasks containing sterile AA medium, a rapid decrease in the concentration of hCTLA4Ig, independent on pH, was observed resulting in more than 90% of the protein loss within 1 h due to the surface adsorption. When the same experiments were performed on four different types of culture equipments mentioned above, the lowest adsorption level was observed in the plastic flasks and the highest level was observed in the glass flasks. The use of the plastic flasks retarded the adsorptive loss of hCTLA4Ig at the early stage of the protein production. There was a significant increase in the production of hCTLA4Ig when the flasks were coated with bovine serum albumin. However, the spike test of purified hCTLA4Ig at two different concentrations of 15 and 100 mg L(-1) in 500-mL spinner flasks confirmed that the amount of hCTLA4Ig adsorbed was dependent on the surface area of the flasks but not on the concentrations. In conclusion, although the protein adsorption affected the total amount of the protein yielded to some extent, it could be regarded as a minor factor in transgenic plant cell cultures with higher titer.

9 CFR 147.15 - Laboratory procedure recommended for the bacteriological examination of mycoplasma reactors. 12

Code of Federal Regulations, 2013 CFR

2013-01-01

... 10523. (5) Use sterile deionized distilled water to reconstitute penicillin. (6) Sterile serum should be... samples (ml)—5.0 (1:2000) Mycoplasma Broth Base (g)—22.5 Aqueous penicillin (units)—500,000 Sterile serum... Aqueous penicillin (units)—400,000 NAD (ml)—12.5 Cysteine hydrochloride (ml)—12.5 (1) Rotate flask gently...

9 CFR 147.15 - Laboratory procedure recommended for the bacteriological examination of mycoplasma reactors. 12

Code of Federal Regulations, 2012 CFR

2012-01-01

... 10523. (5) Use sterile deionized distilled water to reconstitute penicillin. (6) Sterile serum should be... samples (ml)—5.0 (1:2000) Mycoplasma Broth Base (g)—22.5 Aqueous penicillin (units)—500,000 Sterile serum... Aqueous penicillin (units)—400,000 NAD (ml)—12.5 Cysteine hydrochloride (ml)—12.5 (1) Rotate flask gently...

9 CFR 147.15 - Laboratory procedure recommended for the bacteriological examination of mycoplasma reactors. 12

Code of Federal Regulations, 2014 CFR

2014-01-01

... 10523. (5) Use sterile deionized distilled water to reconstitute penicillin. (6) Sterile serum should be... samples (ml)—5.0 (1:2000) Mycoplasma Broth Base (g)—22.5 Aqueous penicillin (units)—500,000 Sterile serum... Aqueous penicillin (units)—400,000 NAD (ml)—12.5 Cysteine hydrochloride (ml)—12.5 (1) Rotate flask gently...

Gaucher disease: the role of the specialist on metabolic bone diseases.

PubMed

Masi, Laura; Brandi, Maria Luisa

2015-01-01

According to European legislation, a disease can be considered rare or "orphan" when it affects less than 1 subject of 2000 (1). Often these diseases affecting the pediatric age, are complex diseases and chronically debilitating and for this motive need the intervention of multidisciplinary skills specific. Among the rare disease as affecting the skeleton more than 400 are characterized by dysplastic changes of the skeleton (2). Alongside the disorders affecting the skeleton primitively, many systemic diseases can have a bone involvement. Among these, the Gaucher disease (GD), an heterogeneous lysosomal storage determined by hereditary enzyme deficiency of β-glucosidase. Patients with this disease have skeletal disorders of varying severity (Erlenmeyer flask deformity, lytic lesions and osteonecrosis, pathological fractures) that affects both the bone marrow, both mineralized bone with progressive damage of the tissue. The bone disease is the most debilitating of GD and can have a significant impact on the quality of life of patients. Thorough evaluations by monitoring biochemical markers of bone turnover and instrumental, with a quantitative and qualitative evaluation of the bone, are of fundamental importance to intervene early so they can prevent complications irreversible.

Improving cellulase productivity of Penicillium oxalicum RE-10 by repeated fed-batch fermentation strategy.

PubMed

Han, Xiaolong; Song, Wenxia; Liu, Guodong; Li, Zhonghai; Yang, Piao; Qu, Yinbo

2017-03-01

Medium optimization and repeated fed-batch fermentation were performed to improve the cellulase productivity by P. oxalicum RE-10 in submerged fermentation. First, Plackett-Burman design (PBD) and central composite design (CCD) were used to optimize the medium for cellulase production. PBD demonstrated wheat bran and NaNO 3 had significant influences on cellulase production. The CCD results showed the maximum filter paper activity (FPA) production of 8.61U/mL could be achieved in Erlenmeyer flasks. The maximal FPA reached 12.69U/mL by submerged batch fermentation in a 7.5-L stirred tank, 1.76-fold higher than that on the original medium. Then, the repeated fed-batch fermentation strategy was performed successfully for increasing the cellulase productivity from 105.75U/L/h in batch fermentation to 158.38U/L/h. The cellulase activity and the glucan conversion of delignined corn cob residue hydrolysis had no significant difference between the enzymes sampled from different cycles of the repeated fed-batch fermentation and that from batch culture. Copyright © 2016 Elsevier Ltd. All rights reserved.

Laccase production by free and immobilized mycelia of Peniophora cinerea and Trametes versicolor: a comparative study.

PubMed

Silvério, Sara C; Moreira, Sérgio; Milagres, Adriane M F; Macedo, Eugénia A; Teixeira, José A; Mussatto, Solange I

2013-03-01

The production of laccase by immobilized mycelia of Peniophora cinerea and Trametes versicolor was studied. In an initial stage, experimental assays were performed in Erlenmeyer flasks using free and immobilized mycelium, and the performance of the fungal strains to produce the enzyme was compared. Both fungi adhered into the support material (a synthetic fiber), growing not only on the surface but also in the interspaces of the fibers. Immobilization of P. cinerea provided a 35-fold increase in laccase production when compared to the production obtained by using free mycelium. On the other hand, immobilization of T. versicolor caused a decrease in laccase activity. A comparison between the strains revealed that immobilized P. cinerea (3,500 U/L) surpassed the enzyme production by free T. versicolor (800 U/L). When the conditions that gave the best laccase production to each fungus were employed in a stirred tank bioreactor, very low laccase production was observed for both the cases, suggesting that shear stress and mycelia damage caused by the agitation impellers negatively affected the enzyme production.

Can the Short-Term Toxicity of Water-Soluble Jet Fuel Hydrocarbons Produce Long-Lasting Effects in Lake Plankton Communities?

DTIC Science & Technology

1983-09-01

Richard Starr Algal collection or Ankistrodesmus sp. or Chlorella sp. from the Carolina Biological collection. The cultures were unialgal but not... Chlorella were grown in BBM medium. After five days ot growth, half ot the flasks (3) for each species were spiked with 0.1 ml of toluene. The , .. flasks...quickly recovered and grew at rates equa’ the = toluene-free cells, finally reaching the same final biomass. Ankistrodesmus and Chlorella however

Effects of different fermentation methods on bacterial cellulose and acid production by Gluconacetobacter xylinus in Cantonese-style rice vinegar.

PubMed

Fu, Liang; Chen, Siqian; Yi, Jiulong; Hou, Zongxia

2014-07-01

A strain of acidogenic bacterium was isolated from the fermentation liquid of Cantonese-style rice vinegar produced by traditional surface fermentation. 16S rDNA identification confirmed the bacterium as Gluconacetobacter xylinus, which synthesizes bacterial cellulose, and the acid productivity of the strain was investigated. In the study, the effects of the membrane integrity and the comparison of the air-liquid interface membrane with immerged membrane on total acidity, cellulose production, alcohol dehydrogenase (ADH) activity and number of bacteria were investigated. The cellulose membrane and the bacteria were observed under SEM for discussing their relationship. The correlations between oxygen consumption and total acid production rate were compared in surface and shake flask fermentation. The results showed the average acid productivity of the strain was 0.02g/(100mL/h), and the integrity of cellulose membrane in surface fermentation had an important effect on total acidity and cellulose production. With a higher membrane integrity, the total acidity after 144 h of fermentation was 3.75 g/100 mL, and the cellulose production was 1.71 g/100 mL after 360 h of fermentation. However, when the membrane was crushed by mechanical force, the total acidity and the cellulose production were as low as 0.36 g/100 mL and 0.14 g/100 mL, respectively. When the cellulose membrane was forced under the surface of fermentation liquid, the total acid production rate was extremely low, but the activity of ADH in the cellulose membrane was basically the same with the one above the liquid surface. The bacteria were mainly distributed in the cellulose membrane during the fermentation. The bacterial counts in surface fermentation were more than in the shake flask fermentation and G. xylinus consumed the substrate faster, in surface fermentation than in shake flask fermentation. The oxygen consumption rate and total acid production rate of surface fermentation were respectively 26.13 times and 2.92 times that of shake flask fermentation.

Combustion Product Evaluation of Various Charge Sizes and Propellant Formulations. Task 1 Report: Sampling and Analytical Procedures Proposed for Use in Gun Propellant Combustion Product Characterization

DTIC Science & Technology

1988-01-01

ml, 0.01N, NaOH + 1 ml 30% H202 H2S 10 ml. 0.01M CdSO4 + STRactan 10 in H2 Aldehydes 10 ml of 3.1 WM/ml 2.4- dinitrophenylhydrazine (DNPH) soln. in...3 15 IITRI C06673-TR1 I,ý 3. 3.1.2.6 Aldehydes I After sampling, the 11 flasks will be injected with 10 mi of a 3.1 14/mX -2,4- dinitrophenylhydrazine

Thermal Transitions and Reaction Kinetics of Polyhederal Silsesquioxane Containing Phenylethynylphthalimides (Preprint)

DTIC Science & Technology

2010-03-18

Grignard reagent was added drop-wise to a well-stirred THF solution (10 mL) of trichloromethylsilane (5.431 g, 36.3 mmol) in a 250 mL flask. After...chloromethylsilane) The Grignard reagent , (Me3Si)2NC6H4MgBr was prepared exactly as previously described from 37.6 mmol of magnesium and 30 mmol of (N...trimethylsilyl)2-4-bromoaniline in THF (25 mL). This Grignard reagent was added drop-wise to a well-stirred THF solution (10 mL) of trichloromethylsilane

Vitality Stains and Real Time PCR Studies to Delineate the Interactions of Pichia anomala and Aspergillus flavus

USDA-ARS?s Scientific Manuscript database

The objectives of this study were to probe the effect of the yeast, P. anomala against A flavus by using real time RT-PCR technique and vitality fluorescent stains. Yeast and fungi were inoculated into a 250 ml-flask containing 50 ml potato dextrose broth (PDB) at yeast to fungus (Y : F) ratios of ...

Optimization of culture conditions for penicilazaphilone C production by a marine-derived fungus Penicillium sclerotiorum M-22.

PubMed

Zhao, H-G; Wang, M; Lin, Y-Y; Zhou, S-L

2018-03-01

The aim of this study was to optimize the culture conditions of a marine-derived fungus Penicillium sclerotiorum M-22 for the production of penicilazaphilone C (PAC), a novel azaphilonidal derivative exhibiting broad cytotoxic and antibacterial effects. By single factor experiments, the effects to the production of PAC of aged seawater concentration, initial pH values, fermentation time, carbon sources, nitrogen sources and inorganic salt sources were investigated individually. Response surface methodology (RSM) analysis was adopted to investigate the interactions between variables and determine the optimal values for maximum PAC production. Evaluation of the experimental results signified that the optimum conditions for maximum production of PAC (19·85 mg l -1 ) in 250 ml Erlenmeyer flask were fermentation time 24·83 days, pH of 7·00, corn meal concentration of 10·72 g l -1 , yeast extract concentration of 4·58 g l -1 , crude sea salt concentration of 20·59 g l -1 . Production under optimized conditions increased to 1·344-fold comparing to its production prior to optimization. The higher PAC production and the penicilazaphilone C -producing marine fungus would be provide a promising alterative approach for industrial and commercial applications. Penicilazaphilone C (PAC) was a novel azaphilonidal derivative which had exhibited selective cytotoxicity and antibacterial activity. To further enhance production of PAC by optimizing fermentation conditions of Penicillium sclerotiorum M-22 would provide a promising alterative approach for industrial and commercial applications. We used the single factor test to determine the key factors which influence the PAC production. Then through the Response surface methodology and Box-Behnken design to determine the best fermentation condition for maximum production of PAC. Through these experimental designs and analysis will help us improve experimental efficiency and save time and materials. © 2017 The Society for Applied Microbiology.

Scale-up of an alkaline protease from Bacillus pumilus MTCC 7514 utilizing fish meal as a sole source of nutrients.

PubMed

Gupta, Rishikesh Kumar; Prasad, Dinesh; Sathesh, Jaykumar; Naidu, Ramachandra Boopathy; Kamini, Numbi Ramudu; Palanivel, Saravanan; Gowthaman, Marichetti Kuppuswami

2012-09-01

Fish meal grades SL1 and SL2 from Sardine (Sardinella longiceps) and NJ from Pink Perch (Nemipterus japonicas) were evaluated as a sole source of carbon and nitrogen in the medium for alkaline protease production by Bacillus pumilus MTCC 7514. The analysis of the fish meal suggests that the carbon and nitrogen contents in fish meal are sufficient to justify its choice as replacement for other nutrients. Protease production increased significantly (4,914 U/ml) in medium containing only fish meal, compared with the basal medium (2,646 U/ml). However, the elimination of inorganic salts from media reduced the protease productivity. In addition, all the three grades of fish meal yielded almost the same amounts of protease when employed as the sole source of carbon and nitrogen. Nevertheless, the best results were observed in fish meal SL1 medium. Furthermore, protease production was enhanced to 6,966 U/ml and 7,047 U/ml on scaling up from flask (4,914 U/ml) to 3.7 and 20 L fermenters, respectively, using fish meal (10 g/l). Similarly, the corresponding improvement in productivities over flask (102.38 U/ml/h) was 193.5 and 195.75 U/ml/h in 3.7 and 20 L fermenters, respectively. The crude protease was found to have dehairing ability in leather processing, which is bound to have great environmental benefits.

A high efficiency stirrer for suspension cell culture with or without microcarriers.

PubMed

de Bruyne, N A

1984-01-01

The stirrer described in this paper employs "Teaspoon stirring" in which a secondary motion is superimposed on the rotation of the liquid. This secondary motion arises from the viscous drag from the wall and bottom of the flask. The culture medium is rotated by a bulb-ended rod suspended from inside the flask cap: the bulb orbits around in a circular trough formed between a central indent in the base and the rounded periphery of the base. There are no bearings and no stagnant areas. The power used to stir 4 flasks each holding 500 ml of medium is less than 2 watts. The speed is electronically controlled by a tachometer and automatically gives a smooth start and stop as well as "Interval stirring" to assist initial attachment of cells.

Life history trait analysis of the entomopathogenic nematode Steinernema feltiae provides the basis for prediction of dauer juvenile yields in monoxenic liquid culture.

PubMed

Addis, Temesgen; Teshome, Asmamaw; Strauch, Olaf; Ehlers, Ralf-Udo

2016-05-01

Entomopathogenic nematodes (Steinernema spp.) are used in integrated pest management to control insect pests in cryptic environments. The nematodes are mass produced in monoxenic liquid culture with their symbiotic bacteria Xenorhabdus spp. For a better understanding of nematode population dynamics, the life history traits (LHTs) of the entomopathogenic nematode Steinernema feltiae were assessed at 25 °C by observing single pairs of male and female nematodes using a hanging drop technique. To investigate the influence of different food supplies on nematode reproduction, the LHTs were assessed with a daily supply of 5 ×, 10 × and 20 × 10(9) cells ml(-1) of the nematode's bacterial symbiont Xenorhabdus bovienii in semi-solid nematode growth gelrite (NGG) medium. Increasing bacterial density had a significant positive influence on the average number of offspring produced, which ranged from 359 to 813 per female. The intrinsic rate of natural increase r m, which ranges from 1.10 to 1.19 day(-1), was neither influenced by the bacterial density, nor was the mean generation time T (5.12-5.25 days) and population doubling time (PDT) (0.64-0.59 days). The average lifespan of reproductive females, which ranged from 6.7 to 7.3 days, was positively correlated with bacterial density. A positive correlation between female body volume and bacterial density was recorded (R = 0.67) as well as a significant positive correlation between female body size and offspring production (R = 0.89) in hanging drops. Whether these data can be used to predict nematode yields in liquid culture was tested. The total female body volume calculated as the average female body volume × total number of parental females per millilitre 3 days after nematode inoculation was positively correlated (R = 0.72) with nematode yields. The total female body volume on process day 3 is thus a good indicator for the estimation of nematode yield at the end of the process (12-15 days post dauer juvenile (DJ) inoculation) in both Erlenmeyer flasks and bioreactors. With a mean deviation of 9467 DJs ml(-1), the error resembles approximately 5 % of the final DJ yields.

Inhibition of bacterial activity in acid mine drainage

NASA Astrophysics Data System (ADS)

Singh, Gurdeep; Bhatnagar, Miss Mridula

1988-12-01

Acid mine drainage water give rise to rapid growth and activity of an iron- and sulphur- oxidizing bacterium Thiobacillus ferrooxidians which greatly accelerate acid producing reactions by oxidation of pyrite material associated with coal and adjoining strata. The role of this bacterium in production of acid mine drainage is described. This study presents the data which demonstrate the inhibitory effect of certain organic acids, sodium benzoate, sodium lauryl sulphate, quarternary ammonium compounds on the growth of the acidophilic aerobic autotroph Thiobacillus ferrooxidians. In each experiment, 10 milli-litres of laboratory developed culture of Thiobacillus ferrooxidians was added to 250 milli-litres Erlenmeyer flask containing 90 milli-litres of 9-k media supplemented with FeSO4 7H2O and organic compounds at various concentrations. Control experiments were also carried out. The treated and untreated (control) samples analysed at various time intervals for Ferrous Iron and pH levels. Results from this investigation showed that some organic acids, sodium benzoate, sodium lauryl sulphate and quarternary ammonium compounds at low concentration (10-2 M, 10-50 ppm concentration levels) are effective bactericides and able to inhibit and reduce the Ferrous Iron oxidation and acidity formation by inhibiting the growth of Thiobacillus ferrooxidians is also discussed and presented

Gaucher disease: the role of the specialist on metabolic bone diseases

PubMed Central

Masi, Laura; Brandi, Maria Luisa

2015-01-01

Summary According to European legislation, a disease can be considered rare or “orphan” when it affects less than 1 subject of 2000 (1). Often these diseases affecting the pediatric age, are complex diseases and chronically debilitating and for this motive need the intervention of multidisciplinary skills specific. Among the rare disease as affecting the skeleton more than 400 are characterized by dysplastic changes of the skeleton (2). Alongside the disorders affecting the skeleton primitively, many systemic diseases can have a bone involvement. Among these, the Gaucher disease (GD), an heterogeneous lysosomal storage determined by hereditary enzyme deficiency of β-glucosidase. Patients with this disease have skeletal disorders of varying severity (Erlenmeyer flask deformity, lytic lesions and osteonecrosis, pathological fractures) that affects both the bone marrow, both mineralized bone with progressive damage of the tissue. The bone disease is the most debilitating of GD and can have a significant impact on the quality of life of patients. Thorough evaluations by monitoring biochemical markers of bone turnover and instrumental, with a quantitative and qualitative evaluation of the bone, are of fundamental importance to intervene early so they can prevent complications irreversible. PMID:26604943

Improving xylitol production at elevated temperature with engineered Kluyveromyces marxianus through over-expressing transporters.

PubMed

Zhang, Jia; Zhang, Biao; Wang, Dongmei; Gao, Xiaolian; Hong, Jiong

2015-01-01

Three transporter genes including Kluyveromyces marxianus aquaglyceroporin gene (KmFPS1), Candida intermedia glucose/xylose facilitator gene (CiGXF1) or glucose/xylose symporter gene (CiGXS1) were over-expressed in K. marxianus YZJ017 to improve xylitol production at elevated temperatures. The xylitol production of YZJ074 that harbored CiGXF1 was improved to 147.62g/L in Erlenmeyer flask at 42°C. In fermenter, 99.29 and 149.60g/L xylitol were produced from 99.55 and 151.91g/L xylose with productivity of 4.14 and 3.40g/L/h respectively at 42°C. Even at 45°C, YZJ074 could produce 101.30g/L xylitol from 101.41g/L xylose with productivity of 2.81g/L/h. Using fed-batch fermentation through repeatedly adding non-sterilized substrate directly, YZJ074 could produce 312.05g/L xylitol which is the highest yield reported to date. The engineered strains YZJ074 which can produce xylitol at elevated temperatures is an excellent foundation for xylitol bioconversion. Copyright © 2014 Elsevier Ltd. All rights reserved.

Development of Synthesis and Large Scale Production Technology for Ultrahigh Energy and Density Fluoro-Organic Compounds

DTIC Science & Technology

2005-12-01

M-I (H2C=CHSF5). To 3.5 g potassium hydroxide in 4 mL of water was added 10.5 mL of isopropanol in a 50 mL three-necked flask equipped with an...S- 5 2006 Ne Tech New Era Technology, Inc. 3720 NW 4 3rd Street, Suite 105, Gainesville, Florida 32606-6190, USA Final Report TITLE: Development of ...Senior Scientist, NeTech Research Institute (Subcontractor): University of Florida Dr. William R. Dolbier, Jr., P1 - University of Florida, Crow

Fabrication of a Mechanically Robust Carbon Nanofiber Foam

DTIC Science & Technology

2015-06-01

Erlenmeyer exhaust trap utilizing zeolite and permanganate . ........................ 11   Figure 9.   Early CFF experimental mold...containing zeolite and permanganate to dilute the exhaust gases and trap unreacted ethylene prior to their release. Figure 7. MKS mass flow...controller (model MKS 647a). Figure 8. Erlenmeyer exhaust trap utilizing zeolite and permanganate . 12 c. Gas Mixture A flow of pure compressed

Synthesis of a Biologically Active Oxazol-5-(4H)-One via an Erlenmeyer-Plo¨chl Reaction

ERIC Educational Resources Information Center

Rodrigues, Catarina A. B.; Martinho, Jose´ M. G.; Afonso, Carlos A. M.

2015-01-01

The synthesis of (Z)-4-(4-nitrobenzylidene)-2- phenyloxazol-5(4"H")-one, which is a potent immunomodulator and tyrosinase inhibitor, is described as an experiment for an upper-division undergraduate organic chemistry laboratory course. This compound is produced via an Erlenmeyer-Plo¨chl reaction in the absence of any additional solvents…

Production of nattokinase by batch and fed-batch culture of Bacillus subtilis.

PubMed

Cho, Young-Han; Song, Jae Yong; Kim, Kyung Mi; Kim, Mi Kyoung; Lee, In Young; Kim, Sang Bum; Kim, Hyeon Shup; Han, Nam Soo; Lee, Bong Hee; Kim, Beom Soo

2010-09-30

Nattokinase was produced by batch and fed-batch culture of Bacillus subtilis in flask and fermentor. Effect of supplementing complex media (peptone, yeast extract, or tryptone) was investigated on the production of nattokinase. In flask culture, the highest cell growth and nattokinase activity were obtained with 50 g/L of peptone supplementation. In this condition, nattokinase activity was 630 unit/ml at 12 h. In batch culture of B. subtilis in fermentor, the highest nattokinase activity of 3400 unit/ml was obtained at 10h with 50 g/L of peptone supplementation. From the batch kinetics data, it was shown that nattokinase production was growth-associated and culture should be harvested before stationary phase for maximum nattokinase production. In fed-batch culture of B. subtilis using pH-stat feeding strategy, cell growth (optical density monitored at 600 nm) increased to ca. 100 at 22 h, which was 2.5 times higher than that in batch culture. The highest nattokinase activity was 7100 unit/ml at 19 h, which was also 2.1 times higher than that in batch culture. Copyright 2010 Elsevier B.V. All rights reserved.

[Isolation, identification and characterization of a diethylstilbestrol-degrading bacterial strain Serratia sp].

PubMed

Xu, Ran-Fang; Sun, Min-Xia; Liu, Juan; Wang, Hong; Li, Xin; Zhu, Xue-Zhu; Ling, Wan-Ting

2014-08-01

Utilizing the diethylstilbestrol (DES)-degrading bacteria to biodegrade DES is a most reliable technique for cleanup of DES pollutants from the environment. However, little information is available heretofore on the isolation of DES-degrading bacteria and their DES removal performance in the environment. A novel bacterium capable of degrading DES was isolated from the activated sludge of a wastewater treatment plant. According to its morphology, physiochemical characteristics, and 16S rDNA sequence analysis, this strain was identified as Serratia sp.. The strain was an aerobic bacterium, and it could degrade 68.3% of DES (50 mg x L(-1)) after culturing for 7 days at 30 degrees C, 150 r x min(-1) in shaking flasks. The optimal conditions for DES biodegradation by the obtained strain were 30 degrees C, 40-60 mg x L(-1) DES, pH 7.0, 5% of inoculation volume, 0 g x L(-1) of added NaCl, and 10 mL of liquid medium volume in 100 mL flask.

Delivery of Epinephrine in the Vapor Phase for the Treatment of Croup.

PubMed

Leung, Kitty; Newth, Christopher J L; Hotz, Justin C; O'Brien, Kevin C; Fink, James B; Coates, Allan L

2016-04-01

The Vapotherm system delivers high humidity to the airway of patients by using semipermeable tubules where heated liquid water is in contact with air. The humidified air is conducted to the patient via a heated tube. Preliminary clinical observations in infants with croup suggested that epinephrine added to the water supplying the humidity was delivered successfully in the vapor phase. The purpose of this study was to evaluate the efficiency of the delivery of epinephrine in the vapor phase and to develop the feasibility criteria for a clinical pilot study. Thirty milligrams of epinephrine in a 1-L bag of sterile water was used as the humidification source for a Vapotherm 2000i. The output of the heated circuit was condensed and collected into a small Erlenmeyer flask via a metal coil while the whole collection system was submerged in an ice slurry to maintain the outflow temperature from the flask between 0°C and 2°C. The in vitro system was tested at 40°C with flows of 5, 10, and 15 L/min and L-epinephrine concentrations of 15, 30, and 60 mg/L. Each test was duplicated at each of the six conditions. Academic children's hospital research laboratory. None. None. The system recovered more than 90% of the water vapor from the fully saturated air at 40°C. The epinephrine concentration recovery quantified by ultraviolet-visible spectrophotometry was 23.9% (27.5-20.4%) (mean and range) of the initial concentration. At flows of 5, 10, and 15 L/min, the delivery of epinephrine would be 1.8, 3.6, and 4.2 μg/min, respectively, which is in the therapeutic range used for parenteral infusion in young children. The Vapotherm system can be used to deliver epinephrine in pharmacological doses to the respiratory system as a vapor and thus as an alternative to droplets by conventional nebulization.

Processing Method for Creating Ultra-Thin Lead Zirconate Titanate (PZT) Films Via Chemical Solution Deposition

DTIC Science & Technology

2008-12-01

n-propoxide and titanium isopropoxide , were measured with a graduated auto pipet and combined with 45 mL of 2-MOE in a 125 mL flask. The solution...nitrogen (N2). This anneal procedure was used to remove trapped hydrogen from the thin film. Following the anneal, a bi-layer of titanium (Ti) and...dioxide Ti titanium 10 NO. OF COPIES ORGANIZATION 1 ADMNSTR ELEC DEFNS TECHL INFO CTR ATTN DTIC OCP 8725 JOHN J KINGMAN RD STE

40 CFR Appendix 5 to Subpart A of... - Determination of Crude Oil Contamination in Non-Aqueous Drilling Fluids by Gas Chromatography...

Code of Federal Regulations, 2011 CFR

2011-07-01

.... 6.2.2Analytical balance—Capable of weighing 0.1 mg. 6.2.3Glassware. 6.2.3.1Disposable pipettes... standard at 62.5 mg/mL in methylene chloride. Prepare a medium concentration SPTM standard at 1.25 mg/mL by transferring 1.0 mL of the 62.5 mg/mL solution into a 50 mL volumetric flask and diluting to the mark with...

Analysis of Structure and Specific Functional Groups Involved in Acetylcholinesterase Catalysis and Inhibition

DTIC Science & Technology

1992-12-15

et al., 1990). 2. SRpodoptera frugiperda (Sf9. Cells were typically grown in 250 mL of medium in a 500-mL spinner flask with slow stirring at 27"C in...reasonably good expression systems in Spodoptera for preparing large quantities of enzyme. The enzymes prepared from the baculovirus-Sjodo tera system were...4Standard Errors) for Wild-Type and Mutant Acetylcholinesterases Expressed in a Baculovirus- Spodoptera System’ enzyme 10’K, (M) Km tl/K. .. t 101K

Submerged Culture of Grifola gargal and G. sordulenta (Higher Basidiomycetes) from Argentina as a Source of Mycelia with Antioxidant Activity.

PubMed

Postemsky, Pablo Daniel; Curvetto, Néstor Raúl

2015-01-01

Submerged culture is an alternative mycelium source for Grifola gargal and G. sordulenta, two rare edible mushrooms related to Grifola frondosa. This work studies their mycelia as a source of antioxidants. The efficient concentrations of methanolic extracts in both radical scavenging (RS) and reducing power (RP) abilities in G. gargal and in G. sordulenta showed a high antioxidant activity. In the experimental design used, the antioxidant activity mainly depended on the culture conditions rather than on the media composition. Irrespective of the basal culture medium, mycelium methanolic extracts of G. sordulenta obtained from culture in Erlenmeyer flasks showed equivalents to ascorbic acid (EQ(AA)) RS-EQ(AA) and RP-EQ(AA) contents higher than the corresponding values obtained with jar cultures. Under stationary cultivation, G. sordulenta produced approximately 50% higher content in both RS-EQ(AA) and RP-EQ(AA) than the medicinal mushroom G. frondosa. Phenolics correlated with RS-EQ(AA) and RP-EQ(AA) in G. gargal and with RP-EQ(AA) in G. sordulenta; besides, thin-layer chromatography showed these compounds to be at least in part related to the RS capacity. It is concluded that G. gargal and G. sordulenta mycelia are excellent sources of antioxidant metabolites.

A method to estimate the biomass of Spirulina platensis cultivated on a solid medium.

PubMed

Pelizer, Lúcia Helena; Moraes, Iracema de Oliveira

2014-01-01

This paper presents a method to estimate the biomass of Spirulina cultivated on solid medium with sugarcane bagasse as a support, in view of the difficulty in determining biomass concentrations in bioprocesses, particularly those conducted in semi-solid or solid media. The genus Spirulina of the family Oscillatoriaceae comprises the group of multicellular filamentous cyanobacteria (blue-green microalgae). Spirulina is used as fish feed in aquaculture, as a food supplement, a source of vitamins, pigments, antioxidants and fatty acids. Therefore, its growth parameters are extremely important in studies of the development and optimization of bioprocesses. For studies of biomass growth, Spirulina platensis was cultured on solid medium using sugarcane bagasse as a support. The biomass thus produced was estimated by determining the protein content of the material grown during the process, based on the ratio of dry weight to protein content obtained in the surface growth experiments. The protein content of the biomass grown in Erlenmeyer flasks on surface medium was examined daily to check the influence of culture time on the protein content of the biomass. The biomass showed an average protein content of 42.2%. This methodology enabled the concentration of biomass adhering to the sugarcane bagasse to be estimated from the indirect measurement of the protein content associated with cell growth.

Industrial wastewater as raw material for exopolysaccharide production by Rhizobium leguminosarum

PubMed Central

Sellami, Mohamed; Oszako, Tomasz; Miled, Nabil; Ben Rebah, Faouzi

2015-01-01

The objective of this study was to evaluate the exopolysaccharide (EPS) production by Rhizobium leguminosarum cultivated in wastewater generated by oil companies (WWOC1 and WWOC2) and fish processing industry (WWFP). The results obtained in Erlenmeyer flasks indicated that the rhizobial strain grew well in industrial wastewater. Generally, wastewater composition affected the growth and the EPS production. WWFP allowed good bacterial growth similar to that obtained with the standard medium (YMB). During growth, various quantities of EPS were produced and yields varied depending on the media. Growing in YMB, EPS production did not exceed 9.7 g/L obtained after 72 h of growth. In wastewater, the maximum EPS value reached 11.1 g/L obtained with the fish processing wastewater, after 72 h of growth. The use of a mixture of the oil company wastewater (WWOC2) and the fish processing wastewater (WWFP) as culture medium affected not only the rhizobial strain growth, but also EPS production. The highest EPS (42.4 g/L, after 96 h of culture) was obtained using a ratio of WWFP and WWOC2 of 50:50 (v:v). Therefore, this work shows the ability of Rhizobium leguminosarum, growing in industrial wastewater as new economic medium, to produce EPS. This biopolymer could be applied in enormous biotechnological areas. PMID:26273255

Towards Improved Optical Limiters

DTIC Science & Technology

2002-05-03

nitrophthalonitrile, 3.37 g of potassium carbonate, and 0.54 g of neopentyl glycol were added to 10 mL of dry DMSO in a round bottom flask. The... glycol oxygens which in turn replace the nitro groups of the phthalonitrile and form a bond. This substitution reaction produces the bridge unit as

USSR and Eastern Europe Scientific Abstracts, Biomedical and Behavioral Sciences, Number 92

DTIC Science & Technology

1978-07-26

EFFECT OF NUTRIENT MEDIUM COMPOSITION ON THE BIOSYNTHESIS OF CELLULOYTIC ENZYMES WITH SUBMERGED CULTIVATION OF FUNGUS TRICHODERMA LONGIBRACHIATUM 7-26...enzymes using submerged cultivation of Trichoderma longibrachiatum 7-26 was sought. The culture was in a rocker (200 rot/min) in 750 ml flasks at 28-30

Synthesis and Characterization of Furanic Compounds

DTIC Science & Technology

2013-09-01

trifluoroacetic acid, 1.6-M solution of BuLi in hexane, dichloromethane, sodium bicarbonate, pyridinium chlorochromate, potassium permanganate , sodium...intermediate, 2,5-diformylfuran (2.29 g, 18.47 mmol), was oxidized in a 100-mL round-bottom flask with potassium permanganate (4.47 g, 153.1 mmol) in a NaOH

Optimization of flask culture medium and conditions for hyaluronic acid production by a Streptococcus equisimilis mutant nc2168.

PubMed

Chen, Yong-Hao; Li, Jun; Liu, Li; Liu, Hong-Zhi; Wang, Qiang

2012-10-01

A mutant designated NC2168, which was selected from wild-type Streptococcus equisimilis CVCC55116 by ultraviolet ray combined with(60)Co-γ ray treatment and does not produce streptolysin, was employed to produce hyaluronic acid (HA). In order to increase the output of HA in a flask, the culture medium and conditions for NC2168 were optimized in this study. The influence of culture medium ingredients including carbon sources, nitrogen sources and metal ions on HA production was evaluated using factional factorial design. The mathematical model, which represented the effect of each medium component and their interaction on the yield of HA, was established by the quadratic rotary combination design and response surface method. The model estimated that, a maximal yield of HA could be obtained when the concentrations of yeast extract, peptone, glucose, and MgSO4 were set at 3 g/100 mL, 2 g/100 mL, 0.5 g/100 mL and 0.15 g/100 mL, respectively. Compared with the values obtained by other runs in the experimental design, the optimized medium resulted in a remarkable increase in the output of HA and the maximum of the predicted HA production was 174.76 mg/L. The model developed was accurate and reliable for predicting the production of HA by NC2168.Cultivation conditions were optimized by an orthogonal experimental design and the optimal conditions were as follows: temperature 33°C, pH 7.8, agitation speed 200 rpm, medium volume 20 mL.

One-pot bioethanol production from cellulose by co-culture of Acremonium cellulolyticus and Saccharomyces cerevisiae

PubMed Central

2012-01-01

Background While the ethanol production from biomass by consolidated bioprocess (CBP) is considered to be the most ideal process, simultaneous saccharification and fermentation (SSF) is the most appropriate strategy in practice. In this study, one-pot bioethanol production, including cellulase production, saccharification of cellulose, and ethanol production, was investigated for the conversion of biomass to biofuel by co-culture of two different microorganisms such as a hyper cellulase producer, Acremonium cellulolyticus C-1 and an ethanol producer Saccharomyces cerevisiae. Furthermore, the operational conditions of the one-pot process were evaluated for maximizing ethanol concentration from cellulose in a single reactor. Results Ethanol production from cellulose was carried out in one-pot bioethanol production process. A. cellulolyticus C-1 and S. cerevisiae were co-cultured in a single reactor. Cellulase producing-medium supplemented with 2.5 g/l of yeast extract was used for productions of both cellulase and ethanol. Cellulase production was achieved by A. cellulolyticus C-1 using Solka-Floc (SF) as a cellulase-inducing substrate. Subsequently, ethanol was produced with addition of both 10%(v/v) of S. cerevisiae inoculum and SF at the culture time of 60 h. Dissolved oxygen levels were adjusted at higher than 20% during cellulase producing phase and at lower than 10% during ethanol producing phase. Cellulase activity remained 8–12 FPU/ml throughout the one-pot process. When 50–300 g SF/l was used in 500 ml Erlenmeyer flask scale, the ethanol concentration and yield based on initial SF were as 8.7–46.3 g/l and 0.15–0.18 (g ethanol/g SF), respectively. In 3-l fermentor with 50–300 g SF/l, the ethanol concentration and yield were 9.5–35.1 g/l with their yields of 0.12–0.19 (g/g) respectively, demonstrating that the one-pot bioethanol production is a reproducible process in a scale-up bioconversion of cellulose to ethanol. Conclusion A. cellulolyticus cells produce cellulase using SF. Subsequently, the produced cellulase saccharifies the SF, and then liberated reducing sugars are converted to ethanol by S. cerevisiae. These reactions were carried out in the one-pot process with two different microorganisms in a single reactor, which does require neither an addition of extraneous cellulase nor any pretreatment of cellulose. Collectively, the one-pot bioethanol production process with two different microorganisms could be an alternative strategy for a practical bioethanol production using biomass. PMID:22938388

Reduction of C Bonds Proceeds with Retention of Configuration: Stereochemical Investigation of the Heterogeneous Reduction by Dideuterium of (Homohypostrophene)Neopentyl(2-Norbornyl)Platinum(II) Complexes on Platinum Black.

DTIC Science & Technology

1991-04-23

ClIH16: C, 89.12; H, 10.88; Found: C, 88.85; H, 10.90. Synthesis of Grignard Reagents . Neopentylmagnesium chloride. Into a 500-mL round-bottomed flask... Grignard reagent several times using the following procedure. We placed 5.0 g (0.206 mol) of magnesium chips and a magnetic stir bar to a 200-mL round...Norbornylmagnesium bromide. This Grignard reagent was synthesized using a variation on established procedures. 58,60 We transferred under argon 50.0 mL (30.0 mmol

Photolysis of oxyfluorfen in aqueous methanol.

PubMed

Chakraborty, Subhasish K; Chakraborty, Savitri; Bhattacharyya, Anjan; Chowdhury, Ashim

2013-01-01

Photolysis of oxyfluorfen, an herbicide of the nitrodiphenyl ether class, was studied in aqueous methanol under UV and sunlight. UV irradiation was carried out in a borosilicate glass photoreactor (containing 250 ppm oxyfluorfen in 50% aqueous methanol) equipped with a quartz filter and 125 watt mercury lamp (maximum output 254 nm) at 25 ± 1°C. Sunlight irradiation was conducted at 28 ± 1°C in borosilicate Erlenmeyer flasks containing 250 ppm oxyfluorfen in 50% aqueous methanol. The samples from both the irradiated conditions were withdrawn at a definite time interval and extracted to measure oxyfluorfen content by gas chromatography-flame ionization detector for rate study. The half-life values were 20 hours and 2.7 days under UV and sunlight exposure, respectively. Photolysis of oxyfluorfen yielded 13 photoproducts of which three were characterized by infrared spectrophotometer and (1)H NMR and (13)C NMR spectroscopy. The rest of the photoproducts were identified by gas chromatography-mass spectrometry (GC-MS) and thin layer chromatography (TLC). An ionization potential 70 eV was used for electron impact-mass spectrometry (EI-MS) and methane was used as reagent gas for chemical ionization-mass spectrometry (CI-MS). Two of the photoproducts were also synthesized for comparison. The main phototransformation pathways of oxyfluorfen involved nitro reduction, dechlorination, and hydrolysis as well as nucleophiles displacement reaction.

An Efficient In Vitro Propagation Protocol of Cocoyam [Xanthosoma sagittifolium (L) Schott

PubMed Central

Sama, Anne E.; Hughes, Harrison G.; Abbas, Mohamed S.; Shahba, Mohamed A.

2012-01-01

Sprouted corm sections of “South Dade” white cocoyam were potted and maintained in a greenhouse for 8 weeks. Shoot tips of 3–5 mm comprising the apical meristem with 4–6 leaf primordial, and approximately 0.5 mm of corm tissue at the base. These explants were treated to be used into the culture medium. A modified Gamborg's B5 mineral salts supplemented with 0.05 μM 1-naphthaleneacetic acid (NAA) were used throughout the study. Thidiazuron (TDZ) solution containing 0.01% dimethyl sulfoxide (DMSO) was used. Erlenmeyer flasks and test tubes were used for growing cultures. The effect of different media substrate, thidiazuron, and the interaction between TDZ and Benzylaminopurine (BAP) on cocoyam culture were tested. Results indicated that cocoyam can be successfully micropropagated in vitro through various procedures. All concentrations tested (5–20 μM BAP and 1–4 μM TDZ) produced more axillary shoots per shoot tip than the control without cytokinins. Greater proliferation rates were obtained through the use of 20 μM BAP and 2 μM TDZ, respectively, 12 weeks from initiation. Shoots produced with BAP were larger and more normal in appearance than those produced with TDZ, which were small, compressed, and stunted. The use of stationary liquid media is recommended for economic reasons. PMID:22666109

40 CFR Appendix A to Part 80 - Test for the Determination of Phosphorus in Gasoline

Code of Federal Regulations, 2013 CFR

2013-07-01

... Specification for Filter Paper for Use in Chemical Analysis. 3. Summary of method. 3.1 Organic matter in the...) during the entire period of sample heating. Note 1: If the temperature of the hot water bath drops below... 100-ml volumetric flasks submerged to the mark in ice water. 4.4 Filter Paper, for quantitative...

40 CFR Appendix A to Part 80 - Test for the Determination of Phosphorus in Gasoline

Code of Federal Regulations, 2012 CFR

2012-07-01

... Specification for Filter Paper for Use in Chemical Analysis. 3. Summary of method. 3.1 Organic matter in the...) during the entire period of sample heating. Note 1: If the temperature of the hot water bath drops below... 100-ml volumetric flasks submerged to the mark in ice water. 4.4 Filter Paper, for quantitative...

40 CFR Appendix A to Part 80 - Test for the Determination of Phosphorus in Gasoline

Code of Federal Regulations, 2014 CFR

2014-07-01

... Specification for Filter Paper for Use in Chemical Analysis. 3. Summary of method. 3.1 Organic matter in the...) during the entire period of sample heating. Note 1: If the temperature of the hot water bath drops below... 100-ml volumetric flasks submerged to the mark in ice water. 4.4 Filter Paper, for quantitative...

46 CFR Appendix III to Part 150 - Testing Procedures for Determining Exceptions to the Chart

Code of Federal Regulations, 2014 CFR

2014-10-01

... provided with shields. Testing chemicals other than liquids—The procedure outlined below was developed for... test tube to a stand behind a safety shield (in a hood). Carefully add from a dropper 0.5ml of the... mixture. The Dewar flask is equipped with a magnetic stirrer having a stirring bar coated with an inert...

46 CFR Appendix III to Part 150 - Testing Procedures for Determining Exceptions to the Chart

Code of Federal Regulations, 2012 CFR

2012-10-01

... provided with shields. Testing chemicals other than liquids—The procedure outlined below was developed for... test tube to a stand behind a safety shield (in a hood). Carefully add from a dropper 0.5ml of the... mixture. The Dewar flask is equipped with a magnetic stirrer having a stirring bar coated with an inert...

46 CFR Appendix III to Part 150 - Testing Procedures for Determining Exceptions to the Chart

Code of Federal Regulations, 2013 CFR

2013-10-01

... provided with shields. Testing chemicals other than liquids—The procedure outlined below was developed for... test tube to a stand behind a safety shield (in a hood). Carefully add from a dropper 0.5ml of the... mixture. The Dewar flask is equipped with a magnetic stirrer having a stirring bar coated with an inert...

Efficient Manufacturing of Therapeutic Mesenchymal Stromal Cells Using the Quantum Cell Expansion System

PubMed Central

Hanley, Patrick J.; Mei, Zhuyong; Durett, April G.; Cabreira-Harrison, Marie da Graca; Klis, Mariola; Li, Wei; Zhao, Yali; Yang, Bing; Parsha, Kaushik; Mir, Osman; Vahidy, Farhaan; Bloom, Debra; Rice, R. Brent; Hematti, Peiman; Savitz, Sean I; Gee, Adrian P.

2014-01-01

Background The use of bone marrow-derived mesenchymal stromal cells (MSCs) as a cellular therapy for various diseases, such as graft-versus-host-disease, diabetes, ischemic cardiomyopathy, and Crohn's disease has produced promising results in early-phase clinical trials. However, for widespread application and use in later phase studies, manufacture of these cells needs to be cost effective, safe, and reproducible. Current methods of manufacturing in flasks or cell factories are labor-intensive, involve a large number of open procedures, and require prolonged culture times. Methods We evaluated the Quantum Cell Expansion system for the expansion of large numbers of MSCs from unprocessed bone marrow in a functionally closed system and compared the results to a flask-based method currently in clinical trials. Results After only two passages, we were able to expand a mean of 6.6×108 MSCs from 25 mL of bone marrow reproducibly. The mean expansion time was 21 days, and cells obtained were able to differentiate into all three lineages: chondrocytes, osteoblasts, and adipocytes. The Quantum was able to generate the target cell number of 2.0×108 cells in an average of 9-fewer days and in half the number of passages required during flask-based expansion. We estimated the Quantum would involve 133 open procedures versus 54,400 in flasks when manufacturing for a clinical trial. Quantum-expanded MSCs infused into an ischemic stroke rat model were therapeutically active. Discussion The Quantum is a novel method of generating high numbers of MSCs in less time and at lower passages when compared to flasks. In the Quantum, the risk of contamination is substantially reduced due to the substantial decrease in open procedures. PMID:24726657

Influence of intermittent pressure, fluid flow, and mixing on the regenerative properties of articular chondrocytes.

PubMed

Carver, S E; Heath, C A

1999-11-05

Equine articular chondrocytes, embedded within a polyglycolic acid nonwoven mesh, were cultured with various combinations of intermittent pressure, fluid flow, and mixing to examine the effects of different physical stimuli on neochondrogenesis from young cells. The cell/polymer constructs were cultured first in 125 ml spinner flasks for 1, 2, or 4 weeks and then in a perfusion system with intermittent pressure for a total of up to 6 weeks. Additional constructs were either cultured for all 6 weeks in the spinner flasks or for 1 week in spinners followed by 5 weeks in the perfusion system without intermittent pressure. Tissue constructs cultivated for 2 or 4 weeks in spinner flasks followed by perfusion with intermittent pressure had significantly higher concentrations of both sulfated glycosaminoglycan and collagen than constructs cultured entirely in spinners or almost entirely in the pressure/perfusion system. Initial cultivation in the spinner flasks, with turbulent mixing, enhanced both cell attachment and early development of the extracellular matrix. Subsequent culture with perfusion and intermittent pressure appeared to accelerate matrix formation. While the correlation was much stronger in the pressurized constructs, the compressive modulus was directly proportional to the concentration of sulfated glycosaminoglycan in all physically stressed constructs. Constructs that were not stressed beyond the 1-week seeding period lost mechanical integrity upon harvest, suggesting that physical stimulation, particularly with intermittent pressure, of immature tissue constructs during their development may contribute to their ultimate biomechanical functionality. Copyright 1999 John Wiley & Sons, Inc.

Influence of simulated microgravity on the longevity of insect-cell culture

NASA Technical Reports Server (NTRS)

Cowger, N. L.; O'Connor, K. C.; Bivins, J. E.

1997-01-01

Simulated microgravity within the NASA High Aspect Rotating-Wall Vessel (HARV) provides a quiescent environment to culture fragile insect cells. In this vessel, the duration of stationary and death phase for cultures of Spodoptera frugiperda cells was greatly extended over that achieved in shaker-flask controls. For both HARV and control cultures, S. frugiperda cells grew to concentrations in excess of 1 x 10(7) viable cells ml-1 with viabilities greater than 90%. In the HARV, stationary phase was maintained 9-15 days in contrast to 4-5 days in the shaker flask. Furthermore, the rate of cell death was reduced in the HARV by a factor of 20-90 relative to the control culture and was characterized with a death rate constant of 0.01-0.02 day-1. Beginning in the stationary phase and continuing in the death phase, there was a significant decrease in population size in the HARV versus an increase in the shaker flask. This phenomenon could represent cell adaptation to simulated microgravity and/or a change in the ratio of apoptotic to necrotic cells. Differences observed in this research between the HARV and its control were attributed to a reduction in hydrodynamic forces in the microgravity vessel.

Highly Efficient Large-Scale Lentiviral Vector Concentration by Tandem Tangential Flow Filtration

PubMed Central

Cooper, Aaron R.; Patel, Sanjeet; Senadheera, Shantha; Plath, Kathrin; Kohn, Donald B.; Hollis, Roger P.

2014-01-01

Large-scale lentiviral vector (LV) concentration can be inefficient and time consuming, often involving multiple rounds of filtration and centrifugation. This report describes a simpler method using two tangential flow filtration (TFF) steps to concentrate liter-scale volumes of LV supernatant, achieving in excess of 2000-fold concentration in less than 3 hours with very high recovery (>97%). Large volumes of LV supernatant can be produced easily through the use of multi-layer flasks, each having 1720 cm2 surface area and producing ~560 mL of supernatant per flask. Combining the use of such flasks and TFF greatly simplifies large-scale production of LV. As a demonstration, the method is used to produce a very high titer LV (>1010 TU/mL) and transduce primary human CD34+ hematopoietic stem/progenitor cells at high final vector concentrations with no overt toxicity. A complex LV (STEMCCA) for induced pluripotent stem cell generation is also concentrated from low initial titer and used to transduce and reprogram primary human fibroblasts with no overt toxicity. Additionally, a generalized and simple multiplexed real- time PCR assay is described for lentiviral vector titer and copy number determination. PMID:21784103

Automatic dispensing of liquid nitrogen in submilliliter doses

NASA Astrophysics Data System (ADS)

Milner, C. J.

1984-10-01

Well-controlled doses of 0.2 to 0.5 ml of liquid nitrogen are delivered, on electrical signal (not more than once per 5 s), as fills of a miniature bucket raised by an automatic hoist. The bucket is lifted, brimming, from the storage flask and then moved sideways until over the receiver. At this point, a steel ball, which has been resting in and sealing a drain hole in the bucket, is lifted from its seat by a magnet fixed alongside the (now descending) bucket. Design features are outlined: some alternative designs, valving liquid through a short drain tube fixed in the storage flask, are briefly reviewed. In tests the device delivered 74 g (approx. 260 doses) during 63 min, the loss by evaporation meanwhile being 11 g from the bucket, implying a transfer efficiency of 87%. An indirect measure indicated the dose sizes as 354±10 μl approximately.

Ethanol production from xylose with the yeast Pichia stipitis and simultaneous product recovery by gas stripping using a gas-lift loop fermentor with attached side-arm (GLSA).

PubMed

Domínguez, J M; Cao, N; Gong, C S; Tsao, G T

2000-02-05

The bioconversion of xylose into ethanol with the yeast Pichia stipitis CBS 5773 is inhibited when 20 g/L of ethanol are present in the fermentation broth. In order to avoid this limitation, the fermentation was carried out with simultaneous recovery of product by CO(2) stripping. The fermentation was also improved by attaching a side-arm to the main body of a classical gas-lift loop fermentor. This side-arm increases the liquid circulation, mass transfer, and gas distribution, reducing the amount of oxygen in the inlet gas necessary to perform the fermentation of xylose under microaerobic conditions (K(L)a approximately 16 h(-1)). The continuous stripping of ethanol from the fermentation broth in this new bioreactor system allowed the consumption of higher xylose concentrations than using Erlenmeyer shaker flasks, improved significantly the process productivity and provided a clean ethanol solution by using an ice-cooled condenser system. Finally, a fed-batch fermentation was carried out with a K(L)a = 15.8 h(-1). Starting with 248.2 g of xylose, 237.6 g of xylose was consumed to produce 88.1 g of ethanol which represents 72.6% of the theoretical yield (47.2 g/L of ethanol was recovered in the condenser, while 9.6 g/L remained in the fermentation broth). Copyright 2000 John Wiley & Sons, Inc.

Recombinant antibody production by perfusion cultures of rCHO cells in a depth filter perfusion system.

PubMed

Lee, Joon Chul; Chang, Ho Nam; Oh, Duk Jae

2005-01-01

Recombinant Chinese hamster ovary cells, producing recombinant antibody against the human platelet, were cultivated in a depth filter perfusion system (DFPS). When perfusion cultures with working volume of 1 L were operated at perfusion rates of 5/d and 6/d, volumetric antibody productivities reached values 28 and 34 times higher than that of batch suspension culture in Erlenmeyer flasks and 43 and 53 times higher than that of batch culture in a controlled stirred tank reactor, respectively. Perfusion cultures in the DFPS showed stable antibody production over the whole culture period of up to 20 days. In the DFPS, inoculated cells in suspension were entrapped in a few hours within the depth filter matrix by medium circulation and retained there until the void space of the filter matrix was saturated by the cultured cells. After cells in the depth filter matrix reached saturation, overgrown viable cells at a perfusion rate of 5/d or 6/d were continuously collected into waste medium at a density of 2-4 x 10(5) cells/mL, which resulted in stable operation at high perfusion rates, maintaining values of process parameters such as glucose/lactate concentration, pH, and dissolved oxygen concentration. Because the DFPS overcomes most drawbacks observed with conventional perfusion systems, it is preferable to be used as a key culture system to produce monoclonal antibody stably for a long culture period.

Nitrous oxide production during nitrification from organic solid waste under temperature and oxygen conditions.

PubMed

Nag, Mitali; Shimaoka, Takayuki; Komiya, Teppei

2016-11-01

Landfill aeration can accelerate the biological degradation of organic waste and reduce methane production; however, it induces nitrous oxide (N2O), a potent greenhouse gas. Nitrification is one of the pathways of N2O generation as a by-product during aerobic condition. This study was initiated to demonstrate the features of N2O production rate from organic solid waste during nitrification under three different temperatures (20°C, 30°C, and 40°C) and three oxygen concentrations (5%, 10%, and 20%) with high moisture content and high substrates' concentration. The experiment was carried out by batch experiment using Erlenmeyer flasks incubated in a shaking water bath for 72 h. A duplicate experiment was carried out in parallel, with addition of 100 Pa of acetylene as a nitrification inhibitor, to investigate nitrifiers' contribution to N2O production. The production rate of N2O ranged between 0.40 × 10(-3) and 1.14 × 10(-3) mg N/g-DM/h under the experimental conditions of this study. The rate of N2O production at 40°C was higher than at 20°C and 30°C. Nitrification was found to be the dominant pathway of N2O production. It was evaluated that optimization of O2 content is one of the crucial parameters in N2O production that may help to minimize greenhouse gas emissions and N turnover during aeration.

Blood transport method for chromosome analysis of residents living near Semipalatinsk nuclear test site.

PubMed

Rodzi, Mohd; Ihda, Shozo; Yokozeki, Masako; Takeichi, Nobuo; Tanaka, Kimio; Hoshi, Masaharu

2009-12-01

A study was conducted to compare the storage conditions and transportation period for blood samples collected from residents living in areas near the Semipalatinsk nuclear test site (SNTS). Experiments were performed to simulate storage and shipping environments. Phytohaemagglutinin (PHA)-stimulated blood was stored in 15-ml tubes (condition A: current transport method) in the absence or in 50-ml flasks (condition B: previous transport method) in the presence of RPMI-1640 and 20% fetal bovine serum (FBS). Samples were kept refrigerated at 4 degrees C and cell viability was assessed after 3, 8, 12 and 14 days of storage. RPMI-1640, 20% FBS and further PHA were added to blood samples under condition A in 50-ml flasks for culture. Whole-blood samples under condition B were directly incubated without further sub-culturing process, neither media nor PHA were added, to adopt a similar protocol to that employed in the previous transport method. Samples in condition A and condition B were incubated for 48 hr at 37 degrees C and their mitotic index was determined. The results showed that viable lymphocytes were consistent in both storage conditions but the mitotic index was higher in condition A than in condition B. Although further confirmation studies have to be carried out, previous chromosomal studies and the present experiment have shown that PHA-stimulated blood could be stored without culture medium for up to 8 days under condition A. The present results will be useful for cytogenetic analysis of blood samples that have been transported long distances wherever a radiation accident has occurred.

Radiochemical Separation and Quantification of Tritium in Metallic Radwastes Generated from CANDU Type NPP - 13279

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

Ahn, H.J.; Choi, K.C.; Choi, K.S.

2013-07-01

As a destructive quantification method of {sup 3}H in low and intermediate level radwastes, bomb oxidation, sample oxidation, and wet oxidation methods have been introduced. These methods have some merits and demerits in the radiochemical separation of {sup 3}H radionuclides. That is, since the bomb oxidation and sample oxidation methods are techniques using heating at high temperature, the separation methods of the radionuclides are relatively simple. However, since {sup 3}H radionuclide has a property of being diffused deeply into the inside of metals, {sup 3}H which is distributed on the surface of the metals can only be extracted if themore » methods are applied. As an another separation method, the wet oxidation method makes {sup 3}H oxidized with an acidic solution, and extracted completely to an oxidized HTO compound. However, incomplete oxidized {sup 3}H compounds, which are produced by reactions of acidic solutions and metallic radwastes, can be released into the air. Thus, in this study, a wet oxidation method to extract and quantify the {sup 3}H radionuclide from metallic radwastes was established. In particular, a complete extraction method and complete oxidation method of incomplete chemical compounds of {sup 3}H using a Pt catalyst were studied. The radioactivity of {sup 3}H in metallic radwastes is extracted and measured using a wet oxidation method and liquid scintillation counter. Considering the surface dose rate of the sample, the appropriate size of the sample was determined and weighed, and a mixture of oxidants was added to a 200 ml round flask with 3 tubes. The flask was quickly connected to the distilling apparatus. 20 mL of 16 wt% H{sub 2}SO{sub 4} was given into the 200-ml round flask through a dropping funnel while under stirring and refluxing. After dropping, the temperature of the mixture was raised to 96 deg. C and the sample was leached and oxidized by refluxing for 3 hours. At that time, the incomplete oxidized {sup 3}H compounds were completely oxidized using the Pt catalysts and produced a stable HTO compound. After that, about a 20 ml solution was distilled in the separation apparatus, and the distillate was mixed with an ultimagold LLT as a cocktail solution. The solution in the vial was left standing for at least 24 hours. The radioactivity of {sup 3}H was counted directly using a liquid scintillation analyzer (Packard, 2500 TR/AB, Alpha and Beta Liquid Scintillation Analyzer). (authors)« less

Evaluation of secretome of highly efficient lignocellulolytic Penicillium sp. Dal 5 isolated from rhizosphere of conifers.

PubMed

Rai, Rohit; Kaur, Baljit; Singh, Surender; Di Falco, Macros; Tsang, Adrian; Chadha, B S

2016-09-01

Penicillium sp. (Dal 5) isolated from rhizosphere of conifers from Dalhousie (Himachal Pradesh, India) was found to be an efficient cellulolytic strain. The culture under shake flask on CWR (cellulose, wheat bran and rice straw) medium produced appreciably higher levels of endoglucanase (35.69U/ml), β-glucosidase (4.20U/ml), cellobiohydrolase (2.86U/ml), FPase (1.2U/ml) and xylanase (115U/ml) compared to other Penicillium strains reported in literature. The mass spectroscopy analysis of Penicillium sp. Dal 5 secretome identified 108 proteins constituting an array of CAZymes including glycosyl hydrolases (GH) belonging to 24 different families, polysaccharide lyases (PL), carbohydrate esterases (CE), lytic polysaccharide mono-oxygenases (LPMO) in addition to swollenin and a variety of carbohydrate binding modules (CBM) indicating an elaborate genetic potential of this strain for hydrolysis of lignocellulosics. Further, the culture extract was evaluated for hydrolysis of alkali treated rice straw, wheat straw, bagasse and corn cob at 10% substrate loading rate. Copyright © 2016 Elsevier Ltd. All rights reserved.

Methodology for enabling high-throughput simultaneous saccharification and fermentation screening of yeast using solid biomass as a substrate.

PubMed

Elliston, Adam; Wood, Ian P; Soucouri, Marie J; Tantale, Rachelle J; Dicks, Jo; Roberts, Ian N; Waldron, Keith W

2015-01-01

High-throughput (HTP) screening is becoming an increasingly useful tool for collating biological data which would otherwise require the employment of excessive resources. Second generation biofuel production is one such process. HTP screening allows the investigation of large sample sets to be undertaken with increased speed and cost effectiveness. This paper outlines a methodology that will enable solid lignocellulosic substrates to be hydrolyzed and fermented at a 96-well plate scale, facilitating HTP screening of ethanol production, whilst maintaining repeatability similar to that achieved at a larger scale. The results showed that utilizing sheets of biomass of consistent density (handbills), for paper, and slurries of pretreated biomass that could be pipetted allowed standardized and accurate transfers to 96-well plates to be achieved (±3.1 and 1.7%, respectively). Processing these substrates by simultaneous saccharification and fermentation (SSF) at various volumes showed no significant difference on final ethanol yields, either at standard shake flask (200 mL), universal bottle (10 mL) or 96-well plate (1 mL) scales. Substrate concentrations of up to 10% (w/v) were trialed successfully for SSFs at 1 mL volume. The methodology was successfully tested by showing the effects of steam explosion pretreatment on both oilseed rape and wheat straws. This methodology could be used to replace large shake flask reactions with comparatively fast 96-well plate SSF assays allowing for HTP experimentation. Additionally this method is compatible with a number of standardized assay techniques such as simple colorimetric, High-performance liquid chromatography (HPLC) and Nuclear magnetic resonance (NMR) spectroscopy. Furthermore this research has practical uses in the biorefining of biomass substrates for second generation biofuels and novel biobased chemicals by allowing HTP SSF screening, which should allow selected samples to be scaled up or studied in more detail.

Nanoscale Imaging with a Single Quantum Dot

DTIC Science & Technology

2011-12-19

mL round-bottom flask equipped with a condenser , a thermometer and a magnetic stirring bar. After the EG was heated to 160 °C in an oil bath, 0.5 mL...radiatively transferred to the wire’s SPP mode through an electric dipole interaction19. The efficiency of this process scales as the spontaneous emission...Liu, J., Gao, D., Li, H.-F. & Lin, J.-M. Controlled photopolymerization of hydrogel microstructures inside microchannels for bioassays. Lab. Chip 9

Oxygen transfer phenomena in 48-well microtiter plates: determination by optical monitoring of sulfite oxidation and verification by real-time measurement during microbial growth.

PubMed

Kensy, Frank; Zimmermann, Hartmut F; Knabben, Ingo; Anderlei, Tibor; Trauthwein, Harald; Dingerdissen, Uwe; Büchs, Jochen

2005-03-20

Oxygen limitation is one of the most frequent problems associated with the application of shaking bioreactors. The gas-liquid oxygen transfer properties of shaken 48-well microtiter plates (MTPs) were analyzed at different filling volumes, shaking diameters, and shaking frequencies. On the one hand, an optical method based on sulfite oxidation was used as a chemical model system to determine the maximum oxygen transfer capacity (OTR(max)). On the other hand, the Respiration Activity Monitoring System (RAMOS) was applied for online measurement of the oxygen transfer rate (OTR) during growth of the methylotropic yeast Hansenula polymorpha. A proportionality constant between the OTR(max) of the biological system and the OTR(max) of the chemical system were indicated from these data, offering the possibility to transform the whole set of chemical data to biologically relevant conditions. The results exposed "out of phase" shaking conditions at a shaking diameter of 1 mm, which were confirmed by theoretical consideration with the phase number (Ph). At larger shaking diameters (2-50 mm) the oxygen transfer rate in MTPs shaken at high frequencies reached values of up to 0.28 mol/L/h, corresponding to a volumetric mass transfer coefficient (k(L)a) of 1,600 1/h. The specific mass transfer area (a) increases exponentially with the shaking frequency up to values of 2,400 1/m. On the contrary, the mass transfer coefficient (k(L)) is constant at a level of about 0.15 m/h over a wide range of shaking frequencies and shaking diameters. However, at high shaking frequencies, when the complete liquid volume forms a thin film on the cylindric wall of the well, the mass transfer coefficient (k(L)) increases linearly to values of up to 0.76 m/h. Essentially, the present investigation demonstrates that the 48-well plate outperforms the 96-well MTP and shake flasks at widely used operating conditions with respect to oxygen supply. The 48-well plates emerge, therefore, as an excellent alternative for microbial cultivation and expression studies combining the advantages of both the high-throughput 96-well MTP and the classical shaken Erlenmeyer flask.

An effective system to produce smoke solutions from dried plant tissue for seed germination studies1

PubMed Central

Coons, Janice; Coutant, Nancy; Lawrence, Barbara; Finn, Daniel; Finn, Stephanie

2014-01-01

• Premise of the study: An efficient and inexpensive system was developed to produce smoke solutions from plant material to research the influence of water-soluble compounds from smoke on seed germination. • Methods and Results: Smoke solutions (300 mL per batch) were produced by burning small quantities (100–200 g) of dried plant material from a range of species in a bee smoker attached by a heater hose to a side-arm flask. The flask was attached to a vacuum water aspirator, to pull the smoke through the water. The entire apparatus was operated in a laboratory fume hood. • Conclusions: Compared with other smoke solution preparation systems, the system described is easy to assemble and operate, inexpensive to build, and effective at producing smoke solutions from desired species in a small indoor space. Quantitative measurements can be made when using this system, allowing for replication of the process. PMID:25202613

Effect of simulated microgravity and shear stress on microcin B17 production by Escherichia coli and on its excretion into the medium

NASA Technical Reports Server (NTRS)

Fang, A.; Pierson, D. L.; Koenig, D. W.; Mishra, S. K.; Demain, A. L.

1997-01-01

Production of the antibacterial polypeptide microcin B17 (MccB17) by Escherichia coli ZK650 was inhibited by simulated microgravity. The site of MccB17 accumulation was found to be different, depending on whether the organism was grown in shaking flasks or in rotating bioreactors designed to establish a simulated microgravity environment. In flasks, the accumulation was cellular, but in the reactors, virtually all the microcin was found in the medium. The change from a cellular site to an extracellular one was apparently not a function of gravity, since extracellular production occurred in these bioreactors, irrespective of whether they were operated in the simulated microgravity or normal gravity mode. More probably, excretion is due to the much lower degree of shear stress in the bioreactors. Addition of even a single glass bead to the 50-ml medium volume in the bioreactor created enough shear to change the site of MccB17 accumulation from the medium to the cells.

Impact of carbon and nitrogen feeding strategy on high production of biomass and docosahexaenoic acid (DHA) by Schizochytrium sp. LU310.

PubMed

Ling, Xueping; Guo, Jing; Liu, Xiaoting; Zhang, Xia; Wang, Nan; Lu, Yinghua; Ng, I-Son

2015-05-01

A new isolated Schizochytrium sp. LU310 from the mangrove forest of Wenzhou, China, was found as a high producing microalga of docosahexaenoic acid (DHA). In this study, the significant improvements for DHA fermentation by the batch mode in the baffled flasks (i.e. higher oxygen supply) were achieved. By applied the nitrogen-feeding strategy in 1000 mL baffled flasks, the biomass, DHA concentration and DHA productivity were increased by 110.4%, 117.9% and 110.4%, respectively. Moreover, DHA concentration of 21.06 g/L was obtained by feeding 15 g/L of glucose intermittently, which was an increase of 41.25% over that of the batch mode. Finally, an innovative strategy was carried out by intermittent feeding carbon and simultaneously feeding nitrogen. The maximum DHA concentration and DHA productivity in the fed-batch cultivation reached to 24.74 g/L and 241.5 mg/L/h, respectively. Copyright © 2014 Elsevier Ltd. All rights reserved.

Orally Administered Bioadherent Sustained Release Microencapsulated Vaccines

DTIC Science & Technology

1996-09-01

ml flask culture (MMY media) was inoculated with a recombinant Pichia yeast clone containing the vpB gene integrated at the alcohol oxidase (AOX...measured. 6 DAMD17-95-C-5099 2. Materials and Methods 2.1. Materials vpB is expressed in Pichia yeast and isolated and purified as described below. SEA...locus in the Pichia genome. The culture was grown at 30"C for approximately 48 hours until an optical density ( OD 650) of 3.0 was reached. The culture

Feasibility of Biomonitoring of Exposure to Permethrin Through Analysis of Long-Lived (Metabolite) Adducts to Proteins

DTIC Science & Technology

2007-09-01

Preparation of the Grignard reagent To flask E, containing magnesium (52 mg, 2 mmol) and a few crystals of iodine was added a solution of 3...bromodiphenylether (750 mg, 3 mmol) in diethyl ether (3 mL). The entire system was kept under nitrogen. The preparation of the Grignard reagent was initiated by...device represented below was used for the multi-step synthesis of 3-PBA; all reagents were present in the manifold prior to the beginning of the

Production of C4 and C5 branched-chain alcohols by engineered Escherichia. coli.

PubMed

Chen, Xiaoyan; Xu, Jingliang; Yang, Liu; Yuan, Zhenhong; Xiao, Shiyuan; Zhang, Yu; Liang, Cuiyi; He, Minchao; Guo, Ying

2015-11-01

Higher alcohols, longer chain alcohols, contain more than 3 carbon atoms, showed close energy advantages as gasoline, and were considered as the next generation substitution for chemical fuels. Higher alcohol biosynthesis by native microorganisms mainly needs gene expression of heterologous keto acid decarboxylase and alcohol dehydrogenases. In the present study, branched-chain α-keto acid decarboxylase gene from Lactococcus lactis subsp. lactis CICC 6246 (Kivd) and alcohol dehydrogenases gene from Zymomonas mobilis CICC 41465 (AdhB) were transformed into Escherichia coli for higher alcohol production. SDS-PAGE results showed these two proteins were expressed in the recombinant strains. The resulting strain was incubated in LB medium at 37 °C in Erlenmeyer flasks and much more 3-methyl-1-butanol (104 mg/L) than isobutanol (24 mg/L) was produced. However, in 5 g/L glucose-containing medium, the production of two alcohols was similar, 156 and 161 mg/L for C4 (isobutanol) and C5 (3-methyl-1-butanol) alcohol, respectively. Effects of fermentation factors including temperature, glucose content, and α-keto acid on alcohol production were also investigated. The increase of glucose content and the adding of α-keto acids facilitated the production of C4 and C5 alcohols. The enzyme activities of pure Kivd on α-ketoisovalerate and α-ketoisocaproate were 26.77 and 21.24 μmol min(-1) mg(-1), respectively. Due to its ability on decarboxylation of α-ketoisovalerate and α-ketoisocaproate, the recombinant E. coli strain showed potential application on isoamyl alcohol and isobutanol production.

Optimization of trehalose production by a novel strain Brevibacterium sp. SY361.

PubMed

Wang, Lei; Huang, Rui; Gu, Guanbin; Fang, Hongying

2008-10-01

Trehalose production by a novel strain of Brevibacterium sp. SY361 was optimized in submerged fermentation. Different chemical and physical parameters such as carbon and nitrogen sources, inoculum level, initial pH, incubation temperature, aeration and time-course of fermentation, were studied in order to increase trehalose productivity. An optimal production medium containing 3% (w/v) glucose, 0.9% (v/v) corn steep liquor, 0.5% (w/v) KH(2)PO(4) and 0.4% (w/v) MgSO(4).7 H(2)O was found suitable for trehalose production. An optimal volume of medium in a 500 ml flask was 80 ml. The optimal levels of other parameters were 4.0% (v/v) of inoculum, initial pH of 6.0, incubation temperature of 28-32 degrees C and time-course of 60 h. Optimized parameters gave a maximum trehalose of 12.2 mg/ml with a conversion rate of 58.4%. (c) 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Enhanced production of alkaline thermostable keratinolytic protease from calcium alginate immobilized cells of thermoalkalophilic Bacillus halodurans JB 99 exhibiting dehairing activity.

PubMed

Shrinivas, Dengeti; Kumar, Raghwendra; Naik, G R

2012-01-01

The thermoalkalophilic Bacillus halodurans JB 99 cells known for production of novel thermostable alkaline keratinolytic protease were immobilized in calcium alginate matrix. Batch and repeated batch cultivation using calcium alginate immobilized cells were studied for alkaline protease production in submerged fermentation. Immobilized cells with 2.5% alginate and 350 beads/flask of initial cell loading showed enhanced production of alkaline protease by 23.2% (5,275 ± 39.4 U/ml) as compared to free cells (4,280 ± 35.4 U/ml) after 24 h. In the semicontinuous mode of cultivation, immobilized cells under optimized conditions produced an appreciable level of alkaline protease in up to nine cycles and reached a maximal value of 5,975 U/ml after the seventh cycle. The enzyme produced from immobilized cells efficiently degraded chicken feathers in the presence of a reducing agent which can help the poultry industry in the management of keratin-rich waste and obtaining value-added products.

Organo-Metallic Elements for Associative Information Processing

DTIC Science & Technology

1989-01-15

7844 400 MHz oscilloscope Hamamatsu Model B2297-3 Ge photodiodes Scientech Model 365 power meter Miscellaneous optical components (i.e., harmonic...dried in vacuo at 80 °C for 3 hours to give a tan colored powder 234.2 g, yield I 84%, mp > 400 oC. IR(KBr) 1787, 1733, 1683, 1621, 1557, 1467, 1375...flask was cooled in an ice-water bath for 3 hours and the solid was collected on a filter and washed thoroughly with water 2 x 400 mL and ethanol 3 x

The conversion of BTEX compounds by single and defined mixed cultures to medium-chain-length polyhydroxyalkanoate.

PubMed

Nikodinovic, Jasmina; Kenny, Shane T; Babu, Ramesh P; Woods, Trevor; Blau, Werner J; O'Connor, Kevin E

2008-09-01

Here, we report the use of petrochemical aromatic hydrocarbons as a feedstock for the biotechnological conversion into valuable biodegradable plastic polymers--polyhydroxyalkanoates (PHAs). We assessed the ability of the known Pseudomonas putida species that are able to utilize benzene, toluene, ethylbenzene, p-xylene (BTEX) compounds as a sole carbon and energy source for their ability to produce PHA from the single substrates. P. putida F1 is able to accumulate medium-chain-length (mcl) PHA when supplied with toluene, benzene, or ethylbenzene. P. putida mt-2 accumulates mcl-PHA when supplied with toluene or p-xylene. The highest level of PHA accumulated by cultures in shake flask was 26% cell dry weight for P. putida mt-2 supplied with p-xylene. A synthetic mixture of benzene, toluene, ethylbenzene, p-xylene, and styrene (BTEXS) which mimics the aromatic fraction of mixed plastic pyrolysis oil was supplied to a defined mixed culture of P. putida F1, mt-2, and CA-3 in the shake flasks and fermentation experiments. PHA was accumulated to 24% and to 36% of the cell dry weight of the shake flask and fermentation grown cultures respectively. In addition a three-fold higher cell density was achieved with the mixed culture grown in the bioreactor compared to shake flask experiments. A run in the 5-l fermentor resulted in the utilization of 59.6 g (67.5 ml) of the BTEXS mixture and the production of 6 g of mcl-PHA. The monomer composition of PHA accumulated by the mixed culture was the same as that accumulated by single strains supplied with single substrates with 3-hydroxydecanoic acid occurring as the predominant monomer. The purified polymer was partially crystalline with an average molecular weight of 86.9 kDa. It has a thermal degradation temperature of 350 degrees C and a glass transition temperature of -48.5 degrees C.

Application of a fluorometric microplate algal toxicity assay for riverine periphytic algal species.

PubMed

Nagai, Takashi; Taya, Kiyoshi; Annoh, Hirochica; Ishihara, Satoru

2013-08-01

Although riverine periphytic algae attached to riverbed gravel are dominant species in flowing rivers, there is limited toxicity data on them because of the difficulty in cell culture and assays. Moreover, it is well known that sensitivity to pesticides differ markedly among species, and therefore the toxicity data for multiple species need to be efficiently obtained. In this study, we investigated the use of fluorometric microplate toxicity assay for testing periphytic algal species. We selected five candidate test algal species Desmodesmus subspicatus, Achnanthidium minutissimum, Navicula pelliculosa, Nitzschia palea, and Pseudanabaena galeata. The selected species are dominant in the river, include a wide range of taxon, and represent actual species composition. Other additional species were also used to compare the sensitivity and suitability of the microplate assay. A 96-well microplate was used as a test chamber and algal growth was measured by in-vivo fluorescence. Assay conditions using microplate and fluorometric measurement were established, and sensitivities of 3,5-dichlorophenol as a reference substance were assayed. The 50 percent effect concentrations (EC50s) obtained by fluorometric microplate assay and those obtained by conventional Erlenmeyer flask assay conducted in this study were consistent. Moreover, the EC50 values of 3,5-dichlorophenol were within the reported confidence intervals in literature. These results supported the validity of our microplate assay. Species sensitivity distribution (SSD) analysis was conducted using the EC50s of five species. The SSD was found to be similar to the SSD obtained using additional tested species, suggesting that SSD using the five species largely represents algal sensitivity. Our results provide a useful and efficient method for high-tier probabilistic ecological risk assessment of pesticides. Copyright © 2013 Elsevier Inc. All rights reserved.

Analysis of sweet diterpene glycosides from Stevia rebaudiana: improved HPLC method.

PubMed

Kolb, N; Herrera, J L; Ferreyra, D J; Uliana, R F

2001-10-01

An improved analytical method was developed which may be applied to quality control of stevioside and rebaudioside A contents in dried leaves of Stevia rebaudiana before processing; in a selective sampling program searching for plants of higher yield in diterpene glycosides content; or when a large number of samples are sent to the laboratory for analysis. The procedure developed involves two steps: solvent extraction followed by an isocratic HPLC analysis. The sample, 1 g of dried leaves of S. rebaudiana, is ground and solvent-extracted with EtOH 70% (w/w) in Erlenmeyer flasks by shaking for 30 min in a 70 degrees C water bath. After the extract was cooled, it was filtered and analyzed by HPLC using an NH(2) column (250 x 4.6 mm) and a mixture of acetonitrile/water (80:20, v/v) as mobile phase, pH 5 adjusted with acetic acid. The detection was in the UV range at 210 nm (0.04 AUFS). Quantitation was performed by means of an external standard calibration curve for each analyte which had been obtained from standard solutions of pure stevioside and rebaudioside A. Working under these conditions there were no observed interference effects. The method saves time in sample preparation, and reduces sample handling and chromatographic analysis time, while having little loss of precision [coefficient of variation (CV%) between 1.8% and 3.0%] and recovery [between 98.5% and 100.5%]. The method was applied to 30 samples of S. rebaudiana from Misiones (Northeastern Argentina), and the stevioside content found ranged between 3.78 and 9.75% (weight) whereas Rebaudioside A content ranged between 1.62 and 7.27% (weight).

Ethanol production from sugarcane bagasse hydrolysate using Pichia stipitis.

PubMed

Canilha, Larissa; Carvalho, Walter; Felipe, Maria das Graças de Almeida; Silva, João Batista de Almeida e; Giulietti, Marco

2010-05-01

The objective of this study was to evaluate the ethanol production from the sugars contained in the sugarcane bagasse hemicellulosic hydrolysate with the yeast Pichia stipitis DSM 3651. The fermentations were carried out in 250-mL Erlenmeyers with 100 mL of medium incubated at 200 rpm and 30 degrees C for 120 h. The medium was composed by raw (non-detoxified) hydrolysate or by hydrolysates detoxified by pH alteration followed by active charcoal adsorption or by adsorption into ion-exchange resins, all of them supplemented with yeast extract (3 g/L), malt extract (3 g/L), and peptone (5 g/L). The initial concentration of cells was 3 g/L. According to the results, the detoxification procedures removed inhibitory compounds from the hemicellulosic hydrolysate and, thus, improved the bioconversion of the sugars into ethanol. The fermentation using the non-detoxified hydrolysate led to 4.9 g/L ethanol in 120 h, with a yield of 0.20 g/g and a productivity of 0.04 g L(-1) h(-1). The detoxification by pH alteration and active charcoal adsorption led to 6.1 g/L ethanol in 48 h, with a yield of 0.30 g/g and a productivity of 0.13 g L(-1) h(-1). The detoxification by adsorption into ion-exchange resins, in turn, provided 7.5 g/L ethanol in 48 h, with a yield of 0.30 g/g and a productivity of 0.16 g L(-1) h(-1).

Microbiological Production of Carotenoids

PubMed Central

Ciegler, Alex; Nelson, George E. N.; Hall, Harlow H.

1962-01-01

Synthesis of β-carotene by mated strains of Blakeslea trispora in shaken-flask culture was considerably enhanced by adding either 5% kerosene after 2 days of fermentation or acid-refined kerosene at the start of fermentation to a grain-based medium that also contained a natural lipid, nonionic detergent, and β-ionone; average yields of 17,500 μg per g of dry fermentation solids (86,000 μg per 100 ml of medium) were attained when refined kerosene was used. Almost all of the carotene was retained within the mycelium. Peak yields were achieved in 5 days. PMID:13879500

Automated simultaneous measurement of the δ(13) C and δ(2) H values of methane and the δ(13) C and δ(18) O values of carbon dioxide in flask air samples using a new multi cryo-trap/gas chromatography/isotope ratio mass spectrometry system.

PubMed

Brand, Willi A; Rothe, Michael; Sperlich, Peter; Strube, Martin; Wendeberg, Magnus

2016-07-15

The isotopic composition of greenhouse gases helps to constrain global budgets and to study sink and source processes. We present a new system for high-precision stable isotope measurements of carbon, hydrogen and oxygen in atmospheric methane and carbon dioxide. The design is intended for analyzing flask air samples from existing sampling programs without the need for extra sample air for methane analysis. CO2 and CH4 isotopes are measured simultaneously using two isotope ratio mass spectrometers, one for the analysis of δ(13) C and δ(18) O values and the second one for δ(2) H values. The inlet carousel delivers air from 16 sample positions (glass flasks 1-5 L and high-pressure cylinders). Three 10-port valves take aliquots from the sample stream. CH4 from 100-mL air aliquots is preconcentrated in 0.8-mL sample loops using a new cryo-trap system. A precisely calibrated working reference air is used in parallel with the sample according to the Principle of Identical Treatment. It takes about 36 hours for a fully calibrated analysis of a complete carousel including extractions of four working reference and one quality control reference air. Long-term precision values, as obtained from the quality control reference gas since 2012, account for 0.04 ‰ (δ(13) C values of CO2 ), 0.07 ‰ (δ(18) O values of CO2 ), 0.11 ‰ (δ(13) C values of CH4 ) and 1.0 ‰ (δ(2) H values of CH4 ). Within a single day, the system exhibits a typical methane δ(13) C standard deviation (1σ) of 0.06 ‰ for 10 repeated measurements. The system has been in routine operation at the MPI-BGC since 2012. Consistency of the data and compatibility with results from other laboratories at a high precision level are of utmost importance. A high sample throughput and reliability of operation are important achievements of the presented system to cope with the large number of air samples to be analyzed. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

Osteoarthritic human chondrocytes proliferate in 3D co-culture with mesenchymal stem cells in suspension bioreactors.

PubMed

Khurshid, Madiha; Mulet-Sierra, Aillette; Adesida, Adetola; Sen, Arindom

2018-03-01

Osteoarthritis (OA) is a painful disease, characterized by progressive surface erosion of articular cartilage. The use of human articular chondrocytes (hACs) sourced from OA patients has been proposed as a potential therapy for cartilage repair, but this approach is limited by the lack of scalable methods to produce clinically relevant quantities of cartilage-generating cells. Previous studies in static culture have shown that hACs co-cultured with human mesenchymal stem cells (hMSCs) as 3D pellets can upregulate proliferation and generate neocartilage with enhanced functional matrix formation relative to that produced from either cell type alone. However, because static culture flasks are not readily amenable to scale up, scalable suspension bioreactors were investigated to determine if they could support the co-culture of hMSCs and OA hACs under serum-free conditions to facilitate clinical translation of this approach. When hACs and hMSCs (1:3 ratio) were inoculated at 20,000 cells/ml into 125-ml suspension bioreactors and fed weekly, they spontaneously formed 3D aggregates and proliferated, resulting in a 4.75-fold increase over 16 days. Whereas the apparent growth rate was lower than that achieved during co-culture as a 2D monolayer in static culture flasks, bioreactor co-culture as 3D aggregates resulted in a significantly lower collagen I to II mRNA expression ratio and more than double the glycosaminoglycan/DNA content (5.8 vs. 2.5 μg/μg). The proliferation of hMSCs and hACs as 3D aggregates in serum-free suspension culture demonstrates that scalable bioreactors represent an accessible platform capable of supporting the generation of clinical quantities of cells for use in cell-based cartilage repair. Copyright © 2017 John Wiley & Sons, Ltd.

pH-metric solubility. 2: correlation between the acid-base titration and the saturation shake-flask solubility-pH methods.

PubMed

Avdeef, A; Berger, C M; Brownell, C

2000-01-01

The objective of this study was to compare the results of a normal saturation shake-flask method to a new potentiometric acid-base titration method for determining the intrinsic solubility and the solubility-pH profiles of ionizable molecules, and to report the solubility constants determined by the latter technique. The solubility-pH profiles of twelve generic drugs (atenolol, diclofenac.Na, famotidine, flurbiprofen, furosemide, hydrochlorothiazide, ibuprofen, ketoprofen, labetolol.HCl, naproxen, phenytoin, and propranolol.HCl), with solubilities spanning over six orders of magnitude, were determined both by the new pH-metric method and by a traditional approach (24 hr shaking of saturated solutions, followed by filtration, then HPLC assaying with UV detection). The 212 separate saturation shake-flask solubility measurements and those derived from 65 potentiometric titrations agreed well. The analysis produced the correlation equation: log(1/S)titration = -0.063(+/- 0.032) + 1.025(+/- 0.011) log(1/S)shake-flask, s = 0.20, r2 = 0.978. The potentiometrically-derived intrinsic solubilities of the drugs were: atenolol 13.5 mg/mL, diclofenac.Na 0.82 microg/mL, famotidine 1.1 mg/ mL, flurbiprofen 10.6 microg/mL, furosemide 5.9 microg/mL, hydrochlorothiazide 0.70 mg/mL, ibuprofen 49 microg/mL, ketoprofen 118 microg/mL, labetolol.HCl 128 microg/mL, naproxen 14 microg/mL, phenytoin 19 microg/mL, and propranolol.HCl 70 microg/mL. The new potentiometric method was shown to be reliable for determining the solubility-pH profiles of uncharged ionizable drug substances. Its speed compared to conventional equilibrium measurements, its sound theoretical basis, its ability to generate the full solubility-pH profile from a single titration, and its dynamic range (currently estimated to be seven orders of magnitude) make the new pH-metric method an attractive addition to traditional approaches used by preformulation and development scientists. It may be useful even to discovery scientists in critical decision situations (such as calibrating computational prediction methods).

Erythropoietin production from CHO cells grown by continuous culture in a fluidized-bed bioreactor.

PubMed

Wang, M-D; Yang, M; Huzel, N; Butler, M

2002-01-20

A Chinese hamster ovary (CHO) cell line that expresses human erythropoietin (huEPO) was in a 2-L Cytopilot fluidized-bed bioreactor with 400 mL macroporous Cytoline-1 microcarriers and a variable perfusion rate of serum-free and protein-free medium for 48 days. The cell density increased to a maximum of 23 x 10(6) cells/mL, beads on day 27. The EPO concentration increased to 600 U/mL during the early part of the culture period (on day 24) and increased further to 980 U/mL following the addition of a higher concentration of glucose and the addition of sodium butyrate. The EPO concentration was significantly higher (at least 2x than that in a controlled stirred-tank bioreactor, in a spinner flask, or in a stationary T-flask culture. The EPO accumulated to a total production of 28,000 kUnits over the whole culture period. The molecular characteristics of EPO with respect to size and pattern of glycosylation did not change with scale up. The pattern of utilization and production of 18 amino acids was similar in the Cytopilot culture to that in a stationary batch culture in a T-flask. The concentration of ammonia was maintained at a low level (< 2 mM) over the entire culture period. The specific rate of consumption of glucose, as well as the specific rates of production of lactate and ammonia, were constant throughout the culture period indicating a consistent metabolic behavior of the cells in the bioreactor. These results indicate the potential of the Cytopilot bioreactor culture system for the continuous production of a recombinant protein over several weeks. Copyright 2002 John Wiley & Sons, Inc.

Flow Dynamics In Eccentrically Rotating Flasks Used For Dispersant Effectiveness Testing

EPA Science Inventory

The evaluation of dispersant effectiveness used for oil spills is commonly done using tests conducted in laboratory flasks. We used a Hot Wire Anemometer (HWA) to characterize mixing dynamics in the Swirling Flask (SF) and the Baffled Flask (BF), the latter is being considered b...

Electrically conductive connection for an electrode

DOEpatents

Hornack, Thomas R.; Chilko, Robert J.

1986-01-01

An electrically conductive connection for an electrode assembly of an electrolyte cell in which aluminum is produced by electrolysis in a molten salt is described. The electrode assembly comprises an electrode flask and a conductor rod. The flask has a collar above an area of minimum flask diameter. The electrically conductive connection comprises the electrode flask, the conductor rod and a structure bearing against the collar and the conductor rod for pulling the conductor rod into compressive and electrical contact with the flask.

Electrically conductive connection for an electrode

DOEpatents

Hornack, T.R.; Chilko, R.J.

1986-09-02

An electrically conductive connection for an electrode assembly of an electrolyte cell in which aluminum is produced by electrolysis in a molten salt is described. The electrode assembly comprises an electrode flask and a conductor rod. The flask has a collar above an area of minimum flask diameter. The electrically conductive connection comprises the electrode flask, the conductor rod and a structure bearing against the collar and the conductor rod for pulling the conductor rod into compressive and electrical contact with the flask. 2 figs.

Monoclonal antibody production using a new supermacroporous cryogel bioreactor.

PubMed

Nilsang, Suthasinee; Nandakumar, Kutty Selva; Galaev, Igor Yu; Rakshit, Sudip Kumar; Holmdahl, Rikard; Mattiasson, Bo; Kumar, Ashok

2007-01-01

A supermacroporous cryogel bioreactor has been developed to culture hybridoma cells for long-term continuous production of monoclonal antibodies (mAb). Hybridoma clone M2139, secreting antibodies against J1 epitope (GERGAAGIAGPK; amino acids, 551-564) of collagen type II, are immobilized in the porous bed matrix of a cryogel column (10 mL bed volume). The cells got attached to the matrix within 48 h after inoculation and grew as a confluent sheet inside the cryogel matrix. Cells were in the lag phase for 15 days and secreted mAb into the circulation medium. Glucose consumption and lactic acid production were also monitored, and during the exponential phase (approximately 20 days), the hybridoma cell line consumed 0.75 mM day-1 glucose, produced 2.48 mM day-1 lactic acid, and produced 6.5 microg mL-1 day-1 mAb during the exponential phase. The mAb concentration reached 130 microg mL-1 after continuous run of the cryogel column for 36 days. The yield of the mAb after purification was 67.5 mg L-1, which was three times greater than the mAb yield obtained from T-flask batch cultivation. Even after the exchange of medium reservoir, cells in the cryogel column were still active and had relatively stable mAb production for an extended period of time. The bioreactor was operated continuously for 55 days without any contamination. The results from ELISA as well as arthritis experiments demonstrate that the antibodies secreted by cells grown on the cryogel column did not differ from antibodies purified from the cells grown in commercial CL-1000 culture flasks. Thus, supermacroporous cryogels can be useful as a supporting material for productive hybridoma cell culture. Cells were found to be viable inside the porous matrix of the cryogel during the study period and secreted antibodies continuously. The antibodies thus obtained from the cryogel reactor were found to be functionally active in vivo, as demonstrated by their capacity to induce arthritis in mice.

Flask sample measurements for CO2, CH4 and CO using cavity ring-down spectrometry

NASA Astrophysics Data System (ADS)

Wang, J.-L.; Jacobson, G.; Rella, C. W.; Chang, C.-Y.; Liu, I.; Liu, W.-T.; Chew, C.; Ou-Yang, C.-F.; Liao, W.-C.; Chang, C.-C.

2013-08-01

In recent years, cavity ring-down spectrometry (CRDS) has been demonstrated to be a highly sensitive, stable and fast analytical technique for real-time in situ measurements of greenhouse gases. In this study, we propose the technique (which we call flask-CRDS) of analyzing whole air flask samples for CO2, CH4 and CO using a custom gas manifold designed to connect to a CRDS analyzer. Extremely stable measurements of these gases can be achieved over a large pressure range in the flask, from 175 to 760 Torr. The wide pressure range is conducive to flask sample measurement in three ways: (1) flask samples can be collected in low-pressure environments (e.g. high-altitude locations); (2) flask samples can be first analyzed for other trace gases with the remaining low-pressure sample for CRDS analysis of CO2, CH4 and CO; and (3) flask samples can be archived and re-analyzed for validation. The repeatability of this method (1σ of 0.07 ppm for CO2, 0.4 ppb for CH4, and 0.5 ppb for CO) was assessed by analyzing five canisters filled with the same air sample to a pressure of 200 Torr. An inter-comparison of the flask-CRDS data with in-situ CRDS measurements at a high-altitude mountain baseline station revealed excellent agreement, with differences of 0.10 ± 0.09 ppm (1σ) for CO2 and 0.9 ± 1.0 ppb for CH4. This study demonstrated that the flask-CRDS method was not only simple to build and operate but could also perform highly accurate and precise measurements of atmospheric CO2, CH4 and CO in flask samples.

Correlation of WAIS IQ in 10 Pairs of Brothers.

ERIC Educational Resources Information Center

Matarazzo, Joseph D.; And Others

1978-01-01

Pairs of brothers were individually examined with Wechsler Adult Intelligence Scale some 10 months apart by an experienced clinical psychologist unaware of the consanguineous relationship. Correlation of .42 for Full Scale IQ is consistent with median correlation reported by Erlenmeyer-Kimling and Jarvik in their 1963 literature review.…

[Studies for analyzing prohibited ingredients such as tetracaine hydrochloride in cosmetics].

PubMed

Tokunaga, Hiroshi; Takeuchi, Orie; Uchino, Tadashi; Ando, Masanori

2004-01-01

Tetracaine hydrochloride (TH) is nominated as the prohibited ingredients in cosmetics in Japanese Pharmaceutical Affairs Act. So the analytical method for TH was investigated by HPLC. After adding 5 ml of TH solution at 10 microg/ml and 2 ml of salicylic acid solution at 75 microg/ml as the internal standard to 0.5 g of the lotion, the mixture was made up to 10 ml with a mixture of water and methanol (1:1) as the testing solution. Milky lotion was procedured as follows: After adding 5 ml of TH solution at 10 microg/ml and 2 ml of internal standard solution to 0.5 g of the milky lotion, the mixture was made up to 10 ml with a mixture of water and methanol (1:1). Two milliliter of this mixture was placed into a centrifuging tube with a cap and 2 ml of hexane was added. After shaking vigorously and centrifuging, the lower layer was used as the testing solution. In the case of the cream, the other procedures were used: 0.5 g of cream was placed into a 10-ml volumetric flask and 1 ml of tetrahydrofuran was added. After dissolving, the mixture of methanol and water (1:1) was added to make up 10.0 ml. Two milliliter of this mixture was placed into a centrifuging tube with a cap and 2.0 ml of hexane was added. After shaking vigorously and centrifuging, the lower layer was used as the testing solution. The testing solution of 20 microl was analyzed by HPLC using the ODS column (CAPCELL PAK C18 column, 4.6 x 250 mm), the mixture of acetonitrile and 50 mmol/l phosphate buffer(pH 2.0)(7:3) and the detection wavelength of 303 nm. The working curves from 0.5 to 6.0 microg/ml showed a linear line between the concentrations of TH and the peak area ratio. There was no interference of peak of TH from the lotion, milky lotion and cream.

Comparative biodegradation of HDPE and LDPE using an indigenously developed microbial consortium.

PubMed

Satlewal, Alok; Soni, Ravindra; Zaidi, Mgh; Shouche, Yogesh; Goel, Reeta

2008-03-01

A variety of bacterial strains were isolated from waste disposal sites of Uttaranchal, India, and some from artificially developed soil beds containing maleic anhydride, glucose, and small pieces of polyethylene. Primary screening of isolates was done based on their ability to utilize high- and low-density polyethylenes (HDPE/LDPE) as a primary carbon source. Thereafter, a consortium was developed using potential strains. Furthermore, a biodegradation assay was carried out in 500-ml flasks containing minimal broth (250 ml) and HDPE/ LDPE at 5 mg/ml concentration. After incubation for two weeks, degraded samples were recovered through filtration and subsequent evaporation. Fourier transform infrared spectroscopy (FTIR) and simultaneous thermogravimetric-differential thermogravimetry-differential thermal analysis TG-DTG-DTA) were used to analyze these samples. Results showed that consortium-treated HDPE (considered to be more inert relative to LDPE) was degraded to a greater extent 22.41% weight loss) in comparison with LDPE (21.70% weight loss), whereas, in the case of untreated samples, weight loss was more for LDPE than HDPE (4.5% and 2.5%, respectively) at 400 degrees . Therefore, this study suggests that polyethylene could be degraded by utilizing microbial consortia in an eco-friendly manner.

Biodegradation of trichloroethylene and toluene by indigenous microbial populations in soil.

PubMed Central

Fan, S; Scow, K M

1993-01-01

The biodegradation of trichloroethylene (TCE) and toluene, incubated separately and in combination, by indigenous microbial populations was measured in three unsaturated soils incubated under aerobic conditions. Sorption and desorption of TCE (0.1 to 10 micrograms ml-1) and toluene (1.0 to 20 micrograms ml-1) were measured in two soils and followed a reversible linear isotherm. At a concentration of 1 micrograms ml-1, TCE was not degraded in the absence of toluene in any of the soils. In combination, both 1 microgram of TCE ml-1 and 20 micrograms of toluene ml-1 were degraded simultaneously after a lag period of approximately 60 to 80 h, and the period of degradation lasted from 70 to 90 h. Usually 60 to 75% of the initial 1 microgram of TCE ml-1 was degraded, whereas 100% of the toluene disappeared. A second addition of 20 micrograms of toluene ml-1 to a flask with residual TCE resulted in another 10 to 20% removal of the chemical. Initial rates of degradation of toluene and TCE were similar at 32, 25, and 18 degrees C; however, the lag period increased with decreasing temperature. There was little difference in degradation of toluene and TCE at soil moisture contents of 16, 25, and 30%, whereas there was no detectable degradation at 5 and 2.5% moisture. The addition of phenol, but not benzoate, stimulated the degradation of TCE in Rindge and Yolo silt loam soils, methanol and ethylene slightly stimulated TCE degradation in Rindge soil, glucose had no effect in either soil, and dissolved organic carbon extracted from soil strongly sorbed TCE but did not affect its rate of biodegradation. PMID:8328806

Enhanced production of ATP-binding cassette protein exporter-dependent lipase by modifying the growth medium components of Pseudomonas fluorescens.

PubMed

Eom, Gyeong Tae; Song, Jae Kwang

2014-08-01

The industrially-important thermostable lipase, TliA, was extracellularly produced in the recombinant Pseudomonas fluorescens by the homologous expression of TliA and its cognate ABC protein exporter, TliDEF. To increase the secretory production of TliA, we optimized the growth temperature and the culture medium of P. fluorescens. The total amount and the specific productivity of lipase was highest at 25 °C of cell growth temperature, although maximal cell growth was observed at 30 °C. Using the culture medium composed of 20 g dextrin l(-1), 40 g Tween 80 l(-1) and 30 g peptone l(-1), TliA was produced at a level of 2,200 U ml(-1) in a flask culture. The TliA production increased about 3.8-fold (8,450 U ml(-1)) in batch fermentation using a 2.5 l fermentor, which was about 7.7-fold higher than that of previously reported TliA production.

Breathing gas perfluorocarbon measurements using an absorber filled with zeolites.

PubMed

Proquitté, H; Rüdiger, M; Wauer, R R; Schmalisch, G

2003-11-01

Perfluorocarbon (PFC) has been widely used in the treatment of respiratory diseases; however, PFC content of the breathing gases remains unknown. Therefore, we developed an absorber using PFC selective zeolites for PFC measurement in gases and investigated its accuracy. To generate a breathing gas with different PFC contents a heated flask was rinsed with a constant air flow of 4 litre x min(-1) and 1, 5, 10, and 20 ml of PFC were infused over 20 min using an infusor. The absorber was placed on an electronic scale and the total PFC volume was calculated from the weight gain. Steady-state increase in weight was achieved 3.5 min after stopping the infusion. The calculated PFC volume was slightly underestimated but the measuring error did not exceed -1% for PFC less than 1 ml. The measurement error decreased with increasing PFC volume. This zeolite absorber is an accurate method to quantitatively determine PFC in breathing gases and can be used as a reference method to validate other PFC sensors.

Preparation of gentiooligosaccharides using Trichoderma viride β-glucosidase.

PubMed

Wang, Fei; Wu, Jing; Chen, Sheng

2018-05-15

The recombinant plasmid pPIC9K-bgl1 containing β-glucosidase bgl1 from Trichoderma viride was constructed by overlapping PCR and integrated into Pichia pastoris KM71. In order to assist the formation of disulfide bonds and thus improve protein folding efficiency, protein disulfide isomerase pdi was co-expressed in the P. pastoris KM71/pPIC9K-bgl1/pPICZ-A-pdi strain, and fermentation in flasks resulted in enzyme activity of 143 U/ml. The enzyme activity of β-glucosidase reached 1402 U/ml following optimisation of fermentation conditions in a 3.6 l bioreactor. With 80% glucose as substrate, gentiooligosaccharides were synthesised by β-glucosidase-based reverse hydrolysis. A yield of 130 g/l was achieved with a conversion rate of 16.25%. With 20% glucose and 40% cellobiose as substrates, gentiooligosaccharides were synthesised by transglycosylation with a yield of 116 g/l and a conversion rate of 19.4%. Copyright © 2017 Elsevier Ltd. All rights reserved.

Identification of significant medium components that affect docosahexaenoic acid production by Schizochytrium sp. SW1

NASA Astrophysics Data System (ADS)

Manikan, Vidyah; Hamid, Aidil A.

2013-11-01

Central composite design (CCD) was employed to investigate the significance of glucose, yeast extract, MSG and sea salt in affecting the amount of docosahexaenoic acid (DHA) accumulated by a locally isolated strain of Schizochytrium. Design Expert software was used to construct a set of experiments where each medium component mentioned above was varied over three levels. Cultivation was carried out in 250mL flasks containing 50mL of medium, incubated at 30°C with 200 rpm agitation for 96 hours. ANOVA was conducted to identify the influential factors and the level of their significance where factors that scored a probability value of less than 0.05 were considered significant. The level of influence for each independent variable was also interpreted using perturbation whereas pattern of interaction between the factors were interpreted using interaction plots. This experiment revealed that yeast extract and monosodium glutamate have significant influence on DHA accumulation process by Schizochytrium sp. SW1.

Cooling of Water in a Flask: Convection Currents in a Fluid with a Density Maximum

ERIC Educational Resources Information Center

Velasco, S.; White, J. A.; Roman, F. L.

2010-01-01

The effect of density inversion on the convective flow of water in a spherical glass flask cooled with the help of an ice-water bath is shown. The experiment was carried out by temperature measurements (cooling curves) taken at three different heights along the vertical diameter of the flask. Flows inside the flask are visualized by seeding the…

A new wireless system for decentralised measurement of physiological parameters from shake flasks

PubMed Central

Vasala, Antti; Panula, Johanna; Bollók, Monika; Illmann, Lutz; Hälsig, Christian; Neubauer, Peter

2006-01-01

Background Shake flasks are widely used because of their low price and simple handling. Many researcher are, however, not aware of the physiological consequences of oxygen limitation and substrate overflow metabolism that occur in shake flasks. Availability of a wireless measuring system brings the possibilities for quality control and design of cultivation conditions. Results Here we present a new wireless solution for the measurement of pH and oxygen from shake flasks with standard sensors, which allows data transmission over a distance of more than 100 metres in laboratory environments. This new system was applied to monitoring of cultivation conditions in shake flasks. The at-time monitoring of the growth conditions became possible by simple means. Here we demonstrate that with typical protocols E. coli shake flask cultures run into severe oxygen limitation and the medium is strongly acidified. Additionally the strength of the new system is demonstrated by continuous monitoring of the oxygen level in methanol-fed Pichia pastoris shake flask cultures, which allows the optimisation of substrate feeding for preventing starvation or methanol overfeed. 40 % higher cell density was obtained by preventing starvation phases which occur in standard shake flask protocols by adding methanol when the respiration activity decreased in the cultures. Conclusion The here introduced wireless system can read parallel sensor data over long distances from shake flasks that are under vigorous shaking in cultivation rooms or closed incubators. The presented technology allows centralised monitoring of decentralised targets. It is useful for the monitoring of pH and dissolved oxygen in shake flask cultures. It is not limited to standard sensors, but can be easily adopted to new types of sensors and measurement places (e.g., new sensor points in large-scale bioreactors). PMID:16504107

Practices of shake-flask culture and advances in monitoring CO2 and O2.

PubMed

Takahashi, Masato; Aoyagi, Hideki

2018-05-01

About 85 years have passed since the shaking culture was devised. Since then, various monitoring devices have been developed to measure culture parameters. O 2 consumed and CO 2 produced by the respiration of cells in shaking cultures are of paramount importance due to their presence in both the culture broth and headspace of shake flask. Monitoring in situ conditions during shake-flask culture is useful for analysing the behaviour of O 2 and CO 2 , which interact according to Henry's law, and is more convenient than conventional sampling that requires interruption of shaking. In situ monitoring devices for shake-flask cultures are classified as direct or the recently developed bypass type. It is important to understand the characteristics of each type along with their unintended effect on shake-flask cultures, in order to improve the existing devices and culture conditions. Technical developments in the bypass monitoring devices are strongly desired in the future. It is also necessary to understand the mechanism underlying conventional shake-flask culture. The existing shaking culture methodology can be expanded into next-generation shake-flask cultures constituting a novel culture environment through a judicious selection of monitoring devices depending on the intended purpose of shake-flask culture. Construction and sharing the databases compatible with the various types of the monitoring devices and measurement instruments adapted for shaking culture can provide a valuable resource for broadening the application of cells with shake-flask culture.

Petroleum residues degradation in laboratory-scale by rhizosphere bacteria isolated from the mangrove ecosystem

NASA Astrophysics Data System (ADS)

Rinanti, A.; Nainggolan, I. J.

2018-01-01

This research is about petroleum bioremediation experiment to obtain bacterial isolate from mangrove ecosystem which potentially degrade petroleum. It was conducted in an Erlenmeyer batch system filled with growth medium of Stone Mineral Salt Solution (SMSS) plus petroleum residue, placed in an incubator shaker with a rotation speed of 120 rpm, temperature 3000C, for 14 research days. Indigenous bacteria that have been isolated and identified from the roots of mangrove plants are Ochrobactrum anthropi and Bacillus sp., Ralstonia pickettii and Bacillus circulans. Those bacteriain both monoculture and consortium form (mixed culture) are incorporated into erlenmeyer as remediator agents. All bacteria can utilize hydrocarbon compounds, but Ralstonia pickettii and Bacillus circulans reached exponential phase faster with more cell count than other bacteria. Compared to single cultures, petroleum degradation by a bacterial consortium provides a higher TPH reduction efficiency, i.e. at 5%, 10%, and 15% of initial TPH of 94.4%, 72%, and 80.3%, respectively. This study proved that all bacteria could optimize hydrocarbon compounds up to 15% TPH load.

Production of a biodegradable plastic-degrading enzyme from cheese whey by the phyllosphere yeast Pseudozyma antarctica GB-4(1)W.

PubMed

Watanabe, Takashi; Shinozaki, Yukiko; Suzuki, Ken; Koitabashi, Motoo; Yoshida, Shigenobu; Sameshima-Yamashita, Yuka; Kuze Kitamoto, Hiroko

2014-08-01

Cheese whey is a by-product of cheese production and has high concentrations of lactose (about 5%) and other nutrients. Pseudozyma antarctica produces a unique cutinase-like enzyme, named PaE, that efficiently degrades biodegradable plastics. A previous study showed that a combination of 1% oil and 0.5% lactose increased cutinase-like enzyme production by another species of yeast. In this study, to produce PaE from cheese whey, we investigated the effects of soybean oil on PaE production (expressed as biodegradable plastic-degrading activity) by P. antarctica growing on lactose or cheese whey. In flask cultures, the final PaE activity was only 0.03 U/ml when soybean oil was used as the sole carbon source, but increased to 1.79 U/ml when a limited amount of soybean oil (under 0.5%) was combined with a relatively high concentration of lactose (6%). Using a 5-L jar fermentor with lactose fed-batch cultivation and periodic soybean oil addition, about 14.6 U/ml of PaE was obtained after 5 days of cultivation. When the lactose was replaced with cheese whey, PaE production was 10.8 U/ml after 3 days of cultivation. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Growth of cultured corneal endothelial cells onto a vitreous carbon matrix.

PubMed

Wickham, M G; Cleveland, P H; Binder, P S; Akers, P H

1983-01-01

Fourth passage cells of a rabbit corneal endothelial line were grown for 1 week in flasks containing pieces of a reticulated vitreous carbon matrix. The rate of cell growth in flasks containing the matrix was consistent with that in control flasks. Small fragments of the vitreous carbon material lying on the flask floor were covered by the monolayers as the cells grew to confluency. Vertical growth of cells onto larger pieces of the matrix proceeded in a staged fashion with maximum cell density on pieces of the matrix closest to the floor of the flask. As defined by scanning electron microscopy, cell growth occurred to a level at least 600 microns above the floor of the flask and the confluent monolayer. This novel culture procedure should be a model situation for study of many different aspects of the in vitro capabilities of corneal endothelial cells.

Limitations and possibilities of green synthesis and long-term stability of colloidal Ag nanoparticles

NASA Astrophysics Data System (ADS)

Velgosová, Oksana; Mražíková, Anna

2017-12-01

In this paper the influence of algae life cycle and the solutions pH on the green synthesis of colloidal Ag nanoparticles (AgNPs) as well as effect of different storage conditions on AgNPs long-term stability was investigated. Silver nanoparticles were biologically synthesized using extracts of Parachlorella kessleri algae cultivated 1, 2, 3 and 4 weeks. The formation of AgNPs was monitored using a UV-vis spectrophotometer and verified by TEM observation. The results confirmed formation of polyhedron and/or near polyhedron AgNPs, ranging between 5 and 60 nm in diameter. The age of algae influenced the synthesis rate and an amount of AgNPs in solution. The best results were obtained using tree weeks old algae. UV-vis analysis and TEM observation also revealed that the size and the stability of AgNPs depend on the pH of solution. AgNPs formed in solutions of higher pH (8 and 10) are polyhedron, fine, with narrow size interval and stabile. Nanoparticles formed in solutions of low pH (2, 4 and 6) started to lose their stability on 10th day of experiment, and the particle size interval was wide. The long-term stability of AgNPs can be influenced by light and temperature conditions. The most significant stability loss was observed at day light and room temperature (21°C). After 200-days significant amount of agglomerated particles settled on the bottom of the Erlenmeyer flask. AgNPs stored at dark and room temperature showed better long-term stability, weak particles agglomeration was observed. AgNPs stored at dark and at temperature 5°C showed the best long-term stability. Such AgNPs remained spherical, fine (5-20 nm), with narrow size interval and stable (no agglomeration) even after more than six months.

Decolorization of textile dyes in an air-lift bioreactor inoculated with Bjerkandera adusta OBR105.

PubMed

Sodaneath, Hong; Lee, Jung-In; Yang, Seung-Ok; Jung, Hyekyeng; Ryu, Hee Wook; Cho, Kyung-Suk

2017-09-19

A new decolorizing white-rot fungus, OBR105, was isolated from Mount Odae in South Korea and identified by the morphological characterization of its fruit body and spores and partial 18s rDNA sequences. The ligninolytic enzyme activity of OBR105 was studied to characterize their decolorizing mechanism using a spectrophotometric enzyme assay. For the evaluation of the decolorization capacity of OBR105, the isolate was incubated in an erlenmeyer flask and in an airlifte bioreator with potato dextrose broth (PDB) medium supplemented with each dye. In addition, the decolorization efficiency of real textile wastewater was evaluated in an airlift bioreactor inoculated with the isolate. The isolate was identified as Bjerkandera adusta and had ligninolytic enzymes such as laccase, lignin peroxidase (LiP), and Mn-dependent peroxidase (MnP). Its LiP activity was higher than its MnP and laccase activities. B. adusta OBR105 successfully decolorized reactive dyes (red 120, blue 4, orange 16, and black 5) and acid dyes (red 114, blue 62, orange 7, and black 172). B. adusta OBR105 decolorized 91-99% of 200 mg L -1 of each dye (except acid orange 7) within 3 days in a PDB medium at 28°C, pH 5, and 150 rpm. This fungus decolorized only 45% of 200 mg L -1 acid orange 7 (single azo-type dye) within 3 days, and the decolorization efficiency did not increase by prolonging the cultivation time. In the air-lift bioreactor, B. adusta OBR105 displayed a high decolorization capacity, greater than 90%, for 3 acid dyes (red 114, blue 62, and black 172) and 1 reactive dye (blue 4) within 10-15 h of treatment. B. adusta OBR105 could decolorize real textile wastewater in the air-lift bioreactor. This result suggests that an air-lift reactor employing B. adusta OBR105 is a promising bioreactor for the treatment of dye wastewater.

Chlorella vulgaris as a lipid source: Cultivation on air and seawater-simulating medium in a helicoidal photobioreactor.

PubMed

Frumento, Davide; Aliakbarian, Bahar; Casazza, Alessandro Alberto; Converti, Attilio; Al Arni, Saleh; da Silva, Milena Fernandes

2016-03-01

The freshwater microalga Chlorella vulgaris was cultured batchwise on the seawater-simulating Schlösser medium either in a 1.1-L-working volume helicoidal photobioreactor (HeP) or Erlenmeyer flask (EF) as control and continuously supplying air as CO2 source. In these systems, maximum biomass concentration reached 1.65 ± 0.17 g L(-1) and 1.25 ± 0.06 g L(-1) , and maximum cell productivity 197.6 ± 20.4 mg L(-1)  day(-1) and 160.8 ± 12.2 mg L(-1)  day(-1) , respectively. Compared to the Bold's Basal medium, commonly employed to cultivate this microorganism on a bench-scale, the Schlösser medium ensured significant increases in all the growth parameters, namely maximum cell concentration (268% in EF and 126% in HeP), maximum biomass productivity (554% in EF and 72% in HeP), average specific growth rate (67% in EF and 42% in HeP), and maximum specific growth rate (233% in EF and 22% in HeP). The lipid fraction of biomass collected at the end of runs was analyzed in terms of both lipid content and fatty acid profile. It was found that the seawater-simulating medium, despite of a 56-63% reduction of the overall biomass lipid content compared to the Bold's Basal one, led in HeP to significant increases in both the glycerides-to-total lipid ratio and polyunsaturated fatty acid content compared to the other conditions taken as an average. These results as a whole suggest that the HeP configuration could be a successful alternative to the present means to cultivate C. vulgaris as a lipid source. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:279-284, 2016. © 2016 American Institute of Chemical Engineers.

The Microbial Degradation of TCE (Trichloroethylene).

DTIC Science & Technology

1987-04-01

enrichment studies . All the sample flasks , including the controls, contained 14C in the 14C02 trap. The 14C measured in the control flask 14C02...layer compared to the controls. These data again suggested that TCE was being biologically modified. Those flasks flushed with air gave the first hard...only slightly soluble in water. All culture flasks were incubated at 250C for a minimum of a week. Results in the carbon and nitrogen source studies are

Atmospheric CO2 Concentrations from the Commonwealth Scientific and Industrial Research Organization (CSIRO) GASLAB Flask Sampling Network (March 1991 - December 2006)

DOE Data Explorer

Steele, L. P. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Aspendale, Victoria, Australia; Krummel, P. B. [Commonwealth Scientific and Industrial Research Organization (CSIRO),; Langenfelds, R. L. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Aspendale, Victoria, Australia

2008-01-01

Individual measurements have been obtained from flask air samples returned to the CSIRO GASLAB. Typical sample storage times range from days to weeks for some sites (e.g. Cape Grim, Aircraft over Tasmania and Bass Strait) to as much as one year for Macquarie Island and the Antarctic sites. Experiments carried out to test for changes in sample CO2 mixing ratio during storage have shown significant drifts in some flask types over test periods of several months to years (Cooper et al., 1999). Corrections derived from the test results are applied to network data according to flask type. These measurements indicate a rise in annual average atmospheric CO2 concentration from 357.72 parts per million by volume (ppmv) in 1992 to 383.05 ppmv in 2006, or an increase in annual average of about 1.81 ppmv/year. These flask data may be compared with other flask measurements from the Scripps Institution of Oceanography, available through 2004 in TRENDS; both indicate an annual average increase of 1.72 ppmv/year throuth 2004. Differences may be attributed to different sampling times or days, different numbers of samples, and different curve-fitting techniques used to obtain monthly and annual average numbers from flask data. Measurement error in flask data is believed to be small (Masarie et al., 2001).

Optimization of Carbon and Nitrogen Sources for Extracellular Polymeric Substances Production by Chryseobacterium indologenes MUT.2.

PubMed

Khani, Mojtaba; Bahrami, Ali; Chegeni, Asma; Ghafari, Mohammad Davoud; Mansouran Zadeh, ALi

2016-06-01

Bacterial Extracellular Polymeric Substances (EPS) are environmental friendly and versatile polymeric materials that are used in a wide range of industries such as: food, textile, cosmetics, and pharmaceuticals. To make the production process of the EPS cost-effective, improvements in the production yield is required which could be implemented through application of processes such as optimized culture conditions, and development of the strains with higher yield ( e.g . through genetic manipulation), or using low-cost substrates. In this work, the effects of carbon and nitrogen sources were studied in order to improve the EPS production by the submerged cultivation of Chryseobacterium indologenes MUT.2. The mesophilic microorganism Chryseobacterium indologenes MUT.2, was grown and maintained in the Luria Bertani agar. The initial basal medium contained: glucose (20 g.L -1 ), yeast extracts (5 g.L -1 ), K 2 HPO 4 (6 g.L -1 ), NaH 2 PO 4 (7 g.L -1 ), NH 4 CL (0.7 g.L -1 ), and MgSO 4 (0.5 g.L -1 ). For evaluating the carbon and nitrogen sources' effect on the fermentation performance, cultures were prepared in 500 mL flasks filled with 300 mL of the medium. The single-factor experiments based on statistics was employed to evaluate and optimize the carbon and nitrogen sources for EPS production in the liquid culture medium of Chryseobacterium indologenes MUT.2. The preferred carbon-sources, sucrose and glucose, commonly gave the highest EPS production of 8.32 and 6.37 g.L -1 , respectively, and the maximum EPS production of 8.87 g.L -1 was achieved when glutamic acid (5 g.L -1 ) was employed as the nitrogen source. In this work, the culture medium for production of EPS by Chryseobacterium indologenes MUT.2 was optimized. Compared to the basal culture medium in shake-flasks and stirred tank bioreactor, the use of optimized culture medium has resulted in a 53% and 73% increase in the EPS production, respectively.

Measurements of evaporated perfluorocarbon during partial liquid ventilation by a zeolite absorber.

PubMed

Proquitté, Hans; Rüdiger, Mario; Wauer, Roland R; Schmalisch, Gerd

2004-01-01

During partial liquid ventilation (PLV) the knowledge of the quantity of exhaled perfluorocarbon (PFC) allows a continuous substitution of the PFC loss to achieve a constant PFC level in the lungs. The aim of our in vitro study was to determine the PFC loss in the mixed expired gas by an absorber and to investigate the effect of the evaporated PFC on ventilatory measurements. To simulate the PFC loss during PLV, a heated flask was rinsed with a constant airflow of 4 L min(-1) and PFC was infused by different speeds (5, 10, 20 mL h(-1)). An absorber filled with PFC selective zeolites was connected with the flask to measure the PFC in the gas. The evaporated PFC volume and the PFC concentration were determined from the weight gain of the absorber measured by an electronic scale. The PFC-dependent volume error of the CO2SMO plus neonatal pneumotachograph was measured by manual movements of a syringe with volumes of 10 and 28 mL with a rate of 30 min(-1). Under steady state conditions there was a strong correlation (r2 = 0.999) between the infusion speed of PFC and the calculated PFC flow rate. The PFC flow rate was slightly underestimated by 4.3% (p < 0.01). However, this bias was independent from PFC infusion rate. The evaporated PFC volume was precisely measured with errors < 1%. The volume error of the CO2SMO-Plus pneumotachograph increased with increasing PFC content for both tidal volumes (p < 0.01). However for PFC flow rates up to 20 mL/h the error of the measured tidal volumes was < 5%. PFC selective zeolites can be used to quantify accurately the evaporated PFC volume during PLV. With increasing PFC concentrations in the exhaled air the measurement errors of ventilatory parameters have to be taken into account.

Increasing efficiency of human mesenchymal stromal cell culture by optimization of microcarrier concentration and design of medium feed.

PubMed

Chen, Allen Kuan-Liang; Chew, Yi Kong; Tan, Hong Yu; Reuveny, Shaul; Weng Oh, Steve Kah

2015-02-01

Large amounts of human mesenchymal stromal cells (MSCs) are needed for clinical cellular therapy. In a previous publication, we described a microcarrier-based process for expansion of MSCs. The present study optimized this process by selecting suitable basal media, microcarrier concentration and feeding regime to achieve higher cell yields and more efficient medium utilization. MSCs were expanded in stirred cultures on Cytodex 3 microcarriers with media containing 10% fetal bovine serum. Process optimization was carried out in spinner flasks. A 2-L bioreactor with an automated feeding system was used to validate the optimized parameters explored in spinner flask cultures. Minimum essential medium-α-based medium supported faster MSC growth on microcarriers than did Dulbecco's modified Eagle's medium (doubling time, 31.6 ± 1.4 vs 42 ± 1.7 h) and shortened the process time. At microcarrier concentration of 8 mg/mL, a high cell concentration of 1.08 × 10(6) cells/mL with confluent cell concentration of 4.7 × 10(4)cells/cm(2) was achieved. Instead of 50% medium exchange every 2 days, we have designed a full medium feed that is based on glucose consumption rate. The optimal medium feed that consisted of 1.5 g/L glucose supported MSC growth to full confluency while achieving the low medium usage efficiency of 3.29 mL/10(6)cells. Finally, a controlled bioreactor with the optimized parameters achieved maximal confluent cell concentration with 16-fold expansion and a further improved medium usage efficiency of 1.68 mL/10(6)cells. We have optimized the microcarrier-based platform for expansion of MSCs that generated high cell yields in a more efficient and cost-effective manner. This study highlighted the critical parameters in the optimization of MSC production process. Copyright © 2015 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

Ecofriendly Synthesis of nano Zero Valent Iron from Banana Peel Extract

NASA Astrophysics Data System (ADS)

Sunardi; Ashadi; Budi Rahardjo, Sentot; Inayati

2017-01-01

In this study, nano Zero Valent Iron (nZVI) were synthesized from banana peel extract (BPE) and ferrous sulfate. During the synthesis of nZVI both the precursor and the reducing agent were mixed in a clean sterilized flask in 1:1 proportion. For the reduction of Fe ions, 5 ml of filtered BPE was mixed to 5 ml of freshly prepared 0.001 M - 0.005 M aqueous of FeSO4 solution with constant stirring at room temperature. Within a particular time change in colour from brown to black color obtained by nanoparticles synthesis. A systematic characterization of nZVI was performed using UV-Vis. UV-visible absorption is used to investigate SPR. Characteristic surface plasmon absorption band was observed at 210 nm for the black colored nZVI synthesized from 0.001-0.005 M ferrous sulfate with BPE concentration 5 ml. It has been found that the optimum concentration for the synthesis of nZVI is 0.001M Fe2+ ions. There is small decrease in the intensity of SPR band from 0.001 to 0.005 M. The characterization size of nZVI was performed using TEM. The result shows that formation of particles size of nZVI was more 100 nm.

Revisiting the round bottom flask rainbow experiment

NASA Astrophysics Data System (ADS)

Selmke, Markus; Selmke, Sarah

2018-01-01

A popular demonstration experiment in optics uses a round-bottom flask filled with water to project a circular rainbow on a screen with a hole through which the flask is illuminated. We show how the vessel's wall shifts the first- and second-order bows towards each other and consequently reduces the width of Alexander's dark band. We address the challenge this introduces in observing Alexander's dark band, and explain the importance of a sufficient distance between the flask and the screen. The wall-effect also introduces a splitting of the bows that can easily be misinterpreted.

Culture expansion of adipose derived stromal cells. A closed automated Quantum Cell Expansion System compared with manual flask-based culture.

PubMed

Haack-Sørensen, Mandana; Follin, Bjarke; Juhl, Morten; Brorsen, Sonja K; Søndergaard, Rebekka H; Kastrup, Jens; Ekblond, Annette

2016-11-16

Adipose derived stromal cells (ASCs) are a rich and convenient source of cells for clinical regenerative therapeutic approaches. However, applications of ASCs often require cell expansion to reach the needed dose. In this study, cultivation of ASCs from stromal vascular fraction (SVF) over two passages in the automated and functionally closed Quantum Cell Expansion System (Quantum system) is compared with traditional manual cultivation. Stromal vascular fraction was isolated from abdominal fat, suspended in α-MEM supplemented with 10% Fetal Bovine Serum and seeded into either T75 flasks or a Quantum system that had been coated with cryoprecipitate. The cultivation of ASCs from SVF was performed in 3 ways: flask to flask; flask to Quantum system; and Quantum system to Quantum system. In all cases, quality controls were conducted for sterility, mycoplasmas, and endotoxins, in addition to the assessment of cell counts, viability, immunophenotype, and differentiation potential. The viability of ASCs passage 0 (P0) and P1 was above 96%, regardless of cultivation in flasks or Quantum system. Expression of surface markers and differentiation potential was consistent with ISCT/IFATS standards for the ASC phenotype. Sterility, mycoplasma, and endotoxin tests were consistently negative. An average of 8.0 × 10 7 SVF cells loaded into a Quantum system yielded 8.96 × 10 7 ASCs P0, while 4.5 × 10 6 SVF cells seeded per T75 flask yielded an average of 2.37 × 10 6 ASCs-less than the number of SVF cells seeded. ASCs P1 expanded in the Quantum system demonstrated a population doubling (PD) around 2.2 regardless of whether P0 was previously cultured in flasks or Quantum, while ASCs P1 in flasks only reached a PD of 1.0. Manufacturing of ASCs in a Quantum system enhances ASC expansion rate and yield significantly relative to manual processing in T-flasks, while maintaining the purity and quality essential to safe and robust cell production. Notably, the use of the Quantum system entails significantly reduced working hours and thereby costs.

Some Interesting Thermodynamics of the Thermos Flask

ERIC Educational Resources Information Center

Berger, Roland

2007-01-01

When opening a thermos flask filled with coffee, one often "hears" the equalization of the pressure difference that appears to be present between the air cavity inside the flask and the surrounding room atmosphere. Recently we discussed this phenomenon while drinking coffee and guessed about the direction of the gas stream accompanying the…

Studies on Viral Disinfection: An Evaluation of Moist Heat Disinfection for HBV by Using A0 Concept Defined in ISO 15883-Washer-Disinfectors.

PubMed

Uetera, Yushi; Kawamura, Kunio; Kobayashi, Hiroyoshi; Saito, Yuhei; Yasuhara, Hiroshi; Saito, Ryoichi

2010-01-01

International Organization for Standardization (ISO) 15883 for washer-disinfectors has introduced the A(0) concept to allow comparison of the lethality of moist heat processes. The A(0) value is the equivalent disinfection time in seconds at 80 °C calculated on the basis of microbial killing kinetics when the disinfection temperature is over 65 °C. Hepatitis B virus (HBV), transmissible only to humans and chimpanzees, is an important heat-resistant, blood-borne pathogen. Therefore, it is mandatory to disinfect HBV thoroughly in the washer-disinfectors employed for surgical instruments. Additionally, it has become extremely difficult to use chimpanzees as experimental models or to perform human volunteer studies. Therefore, it is considered worthwhile to re-evaluate the reported data on the moist heat disinfection of HBV using the A(0) value. In the voluntary active immunization to humans in 1973, HBV serum (infectivity titer: 10(6.5) CID(50)/mL) underwent moist heat disinfection at 98 °C for 1 min in a flask over an electric burner (conservatively estimated A(0) value: 3786). Then, 0.1 mL was inoculated to each of 29 volunteers. No one revealed evidence of infection clinically or in the laboratory tests available at the time. In 1979, a more sensitive test appeared and revealed three sub-clinically infected volunteers. In the 1980s, there were two chimpanzee experimental models using HBV serum (infectivity titer: 10(5) CID(50)/mL). In one model, the serum underwent moist heat disinfection at 98 °C for 2 min in a thermostat bath (conservatively estimated A(0) value: 7571). One milliliter was inoculated to each of two chimpanzees, and both of them revealed no evidence of infection. In another model, the serum underwent moist heat disinfection using two conditions in a thermostat bath, respectively: at 103 °C for 90 s (A(0) value: 24865) and at 65 °C for 10 h (A(0) value: 1138). Ten milliliters of each sample were mixed. Then, the mixture was inoculated to each of two chimpanzees. Both of them revealed no evidence of infection. In the human volunteer study, the serum infectivity titer was more than 30 times (10(1.5) times) higher than that used in the two chimpanzee experimental models. Moreover, the serum was heated in the flask over an electric burner, which is considered less reliable than the thermostat baths to realize uniform heat distribution. It is assumed that these factors were predisposed to the result that a conservatively estimated A(0) value of 3786 failed to inactivate the HBV serum of 10(6.5) CID(50)/mL. In the two chimpanzee models, it was suggested that A(0) value not less than 1138 was able to inactivate the HBV serum of 10(5) CID(50)/mL.

An effective and simplified scale-up strategy for acarbose fermentation based on the carbon source control.

PubMed

Li, Kun-tai; Wie, Sai-jin; Huang, Lin; Cheng, Xin

2012-02-01

The scale-up strategy for acarbose fermentation by Actinoplanes sp. A56 was explored in this paper. The results obtained in shake-flask cultivation demonstrated that the ratio of maltose and glucose had significant effects on the biosynthesis of acarbose, and the feeding medium containing 3:1 (mass ratio) of maltose and glucose was favorable for acarbose production. Then the correlation of the carbon source concentration with acarbose production was further investigated in 100-l fermenter, and the results showed that 7.5-8.0 g of total sugar/100 ml and 4.0-4.5 g of reducing sugar/100 ml were optimal for acarbose production. Based on the results in 100-l fermenter, an effective and simplified scale-up strategy was successfully established for acarbose fermentation in a 30-m(3) fermenter, by using total sugar and reducing sugar as the scale-up parameter. As a result, 4,327 mg of acarbose/l was obtained at 168 h of fermentation.

Cloning and strong expression of a Bacillus subtilis WL-3 mannanase gene in B. subtilis.

PubMed

Yoon, Ki-Hong; Lim, Byung-Lak

2007-10-01

A gene encoding the mannanase of Bacillus subtilis WL-3, which had been isolated from Korean soybean paste, was cloned into Escherichia coli and the nucleotide sequence of a 2.7-kb DNA fragment containing the mannanase gene was subsequently determined. The mannanase gene, designated manA, consisted of 1,080 nucleotides encoding polypeptide of 360 amino acid residues. The deduced amino acid sequence was highly homologous to those of mannanases belonging to glycosyl hydrolase family 26. The manA gene was strongly expressed in B. subtilis 168 by cloning the gene downstream of a strong B. subtilis promoter of plasmid pJ27Delta 88U. In flask cultures, the production of mannanase by recombinant B. subtilis 168 reached maximum levels of 300 units/ml and 450 units/ml in LB medium and LB medium containing 0.3% locust bean gum, respectively. Based on the zymogram of the mannanase, it was found that the mannanase produced by recombinant B. subtilis could be maintained stably without proteolytic degradation during the culture time.

Enhancement of docosahexaenoic acid production by Schizochytrium SW1 using response surface methodology

NASA Astrophysics Data System (ADS)

Nazir, Mohd Yusuf Mohd; Al-Shorgani, Najeeb Kaid Nasser; Kalil, Mohd Sahaid; Hamid, Aidil Abdul

2015-09-01

In this study, three factors (fructose concentration, agitation speed and monosodium glutamate (MSG) concentration) were optimized to enhance DHA production by Schizochytrium SW1 using response surface methodology (RSM). Central composite design was applied as the experimental design and analysis of variance (ANOVA) was used to analyze the data. The experiments were conducted using 500 mL flask with 100 mL working volume at 30°C for 96 hours. ANOVA analysis revealed that the process was adequately represented significantly by the quadratic model (p<0.0001) and two of the factors namely agitation speed and MSG concentration significantly affect DHA production (p<0.005). Level of influence for each variable and quadratic polynomial equation were obtained for DHA production by multiple regression analyses. The estimated optimum conditions for maximizing DHA production by SW1 were 70 g/L fructose, 250 rpm agitation speed and 12 g/L MSG. Consequently, the quadratic model was validated by applying of the estimated optimum conditions, which confirmed the model validity and 52.86% of DHA was produced.

The significance of fructose and MSG in affecting lipid and docosahexaenoic acid (DHA) production of Aurantiochytrium sp. SW1

NASA Astrophysics Data System (ADS)

Rahman, Shariffah Nurhidayah Syed Abdul; Kalil, Mohd Sahaid; Hamid, Aidil Abdul

2018-04-01

Optimization of fermentation medium for the production of docosahexaenoic acid (DHA) by Aurantiochytrium sp. SW1 was carried out. In this study, levels of fructose, monosodium glutamate (MSG) and sea salt were optimized for enhanced lipid and DHA production using response surface methodology (RSM). The design contains a total of 20 runs with 6 central points replication. Cultivation was carried out in 500 mL flasks containing 100 mL nitrogen limited medium at 30°C for 96h. Sequential model sum of squares (SS) revealed that the system was adequately represented by a quadratic model (p<0.0001). ANOVA results showed that fructose and MSG as a single factor has significant positive effect on the DHA content of SW1. The estimated optimal levels of the factors were 100 g/L fructose, 8 g/L MSG and 47% sea salt. Subsequent cultivation employing the suggested values confirmed that the predicted response values were experimentally achievable and reproducible, where 8.82 g/L DHA (51.34% g/g lipid) was achieved.

ARM-LBNL-NOAA Flask Sampler for Carbon Cycle Gases

DOE Data Explorer

Torn, Margaret

2008-01-15

Data from ccg-flasks are sampled at the ARM SGP site and analyzed by the NOAA Earth System Research Laboratory (ESRL) as part of the NOAA Cooperative Global Air Sampling Network. Surface samples are collected from a 60m tower at the SGP Central Facility, usually once per week on one afternoon. The aircraft samples are collected approximately weekly from a chartered aircraft, and the collection flight path is centered over the tower where the surface samples are collected. Samples are collected by the ARM/LBNL Carbon Project. CO2 flask data contains measurements of CO2 concentration and CO2 stable isotope ratios (13CO2 and C18OO) from flasks collected at the SGP site. The flask samples are collected at 2m, 4m, 25m, and 60m along the 60m tower.

Three-dimensional (3D) evaluation of liquid distribution in shake flask using an optical fluorescence technique.

PubMed

Azizan, Amizon; Büchs, Jochen

2017-01-01

Biotechnological development in shake flask necessitates vital engineering parameters e.g. volumetric power input, mixing time, gas liquid mass transfer coefficient, hydromechanical stress and effective shear rate. Determination and optimization of these parameters through experiments are labor-intensive and time-consuming. Computational Fluid Dynamics (CFD) provides the ability to predict and validate these parameters in bioprocess engineering. This work provides ample experimental data which are easily accessible for future validations to represent the hydrodynamics of the fluid flow in the shake flask. A non-invasive measuring technique using an optical fluorescence method was developed for shake flasks containing a fluorescent solution with a waterlike viscosity at varying filling volume (V L  = 15 to 40 mL) and shaking frequency ( n  = 150 to 450 rpm) at a constant shaking diameter (d o  = 25 mm). The method detected the leading edge (LB) and tail of the rotating bulk liquid (TB) relative to the direction of the centrifugal acceleration at varying circumferential heights from the base of the shake flask. The determined LB and TB points were translated into three-dimensional (3D) circumferential liquid distribution plots. The maximum liquid height (H max ) of the bulk liquid increased with increasing filling volume and shaking frequency of the shaking flask, as expected. The toroidal shapes of LB and TB are clearly asymmetrical and the measured TB differed by the elongation of the liquid particularly towards the torus part of the shake flask. The 3D liquid distribution data collected at varying filling volume and shaking frequency, comprising of LB and TB values relative to the direction of the centrifugal acceleration are essential for validating future numerical solutions using CFD to predict vital engineering parameters in shake flask.

CRYOGENIC DEWAR

DOEpatents

Chamberlain, W.H.; Maseck, H.E.

1964-01-28

This patent relates to a dewar for storing cryogenic gase and is of the type having aii inner flask surrounded by a vacuum jacket and having a vent spout through which evaporating gas escapes. Heretofore substantial gas loss has resulted from the radiation of heat towards the flask from the warmer outer elements of the dewar. In this invention, the mask is surrounded by a thermally conducting shield which is disposed in the vacuum space between the flask and the outer elements of the dewar. The shield contacts only the vent spout, which is cooled by the evaporating gas, and thus is maintained at a temperature very close to that of the flask itself. Accordingly, heat radiated toward the flask is intercepted and conducted to the evaporating gas rather than being re-radiated towards the hask. In a liquid helium dewar of typical configniration the mention reduces the boil-off rate by approximately one-half.(AEC)

Carbon and oxygen isotope ratios of ecosystem respiration along an Oregon conifer transect: preliminary observations based on small-flask sampling.

PubMed

Ehleringer, J. R.; Cook, C. S.

1998-01-01

Isotope ratio analyses of atmospheric CO(2) at natural abundance have significant potential for contributing to our understanding of photosynthetic and respiration processes in forest ecosystems. Recent advances in isotope ratio mass spectrometry allow for rapid, on-line analysis of small volumes of CO(2) in air, and open new research opportunities at the ecophysiological, whole-organism, and atmospheric levels. Among the immediate applications are the carbon and oxygen isotope ratio analyses of carbon dioxide in atmospheric air. Routine analysis of carbon dioxide in air volumes of approximately 50-300 &mgr;l is accomplished by linking a commercially available, trace gas condenser and gas chromatograph to an isotope ratio mass spectrometer operated in continuous-flow mode. Samples collected in the field are stored in either gas-tight syringes or 100-ml flasks. The small sample volume required makes it possible to subsample the air in flasks for CO(2) and then to sample the remaining air volume for the analysis of the isotopic composition of either methane or nitrous oxide. Reliable delta(13)C and delta(18)O values can be obtained from samples collected and stored for 1-3 days. Longer-term storage, on the order of weeks, is possible for delta(13)C measurements without drift in the isotope ratio signal, and should also be possible for delta(18)O measurements. When linked with an infrared gas analyzer, pump and flask sampling system, it is feasible to sample CO(2) extensively in remote forest locations. The air-sampling system was used to measure the isotope ratios of atmospheric CO(2) and to conduct a regression analysis of the relationship between these two parameters. From the regression, we calculated the delta(13)C of ecosystem respiration of four coniferous ecosystems along a precipitation gradient in central Oregon. The ecosystems along the coast-to-interior Oregon (OTTER) gradient are dominated by spruce-hemlock forests at the wet, coastal sites (> 200 cm precipitation annually) to juniper woodlands (20 cm precipitation) at the interior, dry end of the transect. The delta(13)C values of ecosystem respiration along this transect differed by only 1.3 per thousand (range of -25.2 to -23.9 per thousand ) during August at the peak of the summer drought. Following autumn rains in September, the delta(13)C of ecosystem respiration in the four stands decreased; overall the difference in the carbon isotope ratio of ecosystem respiration among sites increased to 3.9 per thousand (-26.8 to -22.9 per thousand ).

Determination of Se in biological samples by axial view inductively coupled plasma optical emission spectrometry after digestion with aqua regia and on-line chemical vapor generation

NASA Astrophysics Data System (ADS)

dos Santos, Éder José; Herrmann, Amanda Beatriz; de Caires, Suzete Kulik; Frescura, Vera Lúcia Azzolin; Curtius, Adilson José

2009-06-01

A simple and fast method for the determination of Se in biological samples, including food, by axial view inductively coupled plasma optical emission spectrometry using on-line chemical vapor generation (CVG-ICP OES) is proposed. The concentrations of HCl and NaBH 4, used in the chemical vapor generation were optimized by factorial analysis. Six certified materials (non-fat milk powder, lobster hepatopancreas, human hair, whole egg powder, oyster tissue, and lyophilised pig kidney) were treated with 10 mL of aqua regia in a microwave system under reflux for 15 min followed by additional 15 min in an ultrasonic bath. The solutions were transferred to a 100 mL volumetric flask and the final volume was made up with water. The Se was determined directly in these solutions by CVG-ICP OES, using the analytical line at 196.026 nm. Calibration against aqueous standards in 10% v/v aqua regia in the concentration range of 0.5-10.0 µg L - 1 Se(IV) was used for the analysis. The quantification limit, considering a 0.5 g sample weight in a final volume of 100 mL - 1 was 0.10 µg g - 1. The obtained concentration values were in agreement with the total certified concentrations, according to the t-test for a 95% confidence level.

Dextran Utilization During Its Synthesis by Weissella cibaria RBA12 Can Be Overcome by Fed-Batch Fermentation in a Bioreactor.

PubMed

Baruah, Rwivoo; Deka, Barsha; Kashyap, Niharika; Goyal, Arun

2018-01-01

Weissella cibaria RBA12 produced a maximum of 9 mg/ml dextran (with 90% efficiency) using shake flask culture under the optimized concentration of medium components viz. 2% (w/v) of each sucrose, yeast extract, and K 2 HPO 4 after incubation at optimized conditions of 20 °C and 180 rpm for 24 h. The optimized medium and conditions were used for scale-up of dextran production from Weissella cibaria RBA12 in 2.5-l working volume under batch fermentation in a bioreactor that yielded a maximum of 9.3 mg/ml dextran (with 93% efficiency) at 14 h. After 14 h, dextran produced was utilized by the bacterium till 18 h in its stationary phase under sucrose depleted conditions. Dextran utilization was further studied by fed-batch fermentation using sucrose feed. Dextran on production under fed-batch fermentation in bioreactor gave 35.8 mg/ml after 32 h. In fed-batch mode, there was no decrease in dextran concentration as observed in the batch mode. This showed that the utilization of dextran by Weissella cibaria RBA12 is initiated when there is sucrose depletion and therefore the presence of sucrose can possibly overcome the dextran hydrolysis. This is the first report of utilization of dextran, post-sucrose depletion by Weissella sp. studied in bioreactor.

Development of an ES-like cell culture system (RESC) from rohu, Labeo rohita (Ham.).

PubMed

Goswami, M; Lakra, W S; Yadav, Kamalendra; Jena, J K

2012-12-01

An embryonic stem (ES)-like cell culture system RESC from a commercially important freshwater carp, Labeo rohita, was developed using blastula stage embryos. The cells were cultured in Leibovitz-15 (L-15) medium in gelatin-coated cell culture flask supplemented with 15 % fetal bovine serum along with 10 ng ml(-1) basic fibroblast growth factor at 28 °C under feeder-free conditions. The ES-like cells were characterized by their unique morphology, alkaline phosphatase activity, embryoid body formation tendency, expression of transcription factor Oct4, and consistent chromosome count. The RESC cells when treated with retinoic acid differentiated into cells of different lineages. The RESC developed from mid-blastula embryos of L. rohita would be a useful tool for cellular differentiation and gene expression studies.

Effect of Oxygen-Supply Rates on Growth of Escherichia coli

PubMed Central

McDaniel, L. E.; Bailey, E. G.; Zimmerli, A.

1965-01-01

The effect of oxygen-supply rates on bacterial growth was studied in commercially available unbaffled and baffled flasks with the use of Escherichia coli in a synthetic medium as a test system. The amount of growth obtained depended on the oxygen-supply rate. Based on oxygen-absorption rates (OAR) measured by the rate of sulfite oxidation, equal OAR values in different types of flasks did not give equal amounts of growth. However, growth was essentially equal at the equal sulfite-oxidation rates when these were determined in the presence of killed whole cultures. Specific growth rates were reduced only at oxygen-supply rates much lower than those at which the total amount of growth was reduced. For the physical set-up used in this work and with the biological system employed, Bellco 598 flasks and flasks fitted with Biotech stainless-steel baffles gave satisfactory results at workable broth volumes; unbaffled and Bellco 600 flasks did not. PMID:14264837

Mercury - Its occurrence and economic trends

USGS Publications Warehouse

Bailey, Edgar H.; Smith, Roscoe M.

1964-01-01

price will be a predictable increase in production. If the price averages $300 a flask for 1 year or more, annual production may increase to a rate of 30,000-35,000 flasks in 1966 or 1967. If the price averages $400 a flask, production may increase to 45,000-50,000 flasks a year after a timelag of two or three years. New ore bodies would have to be found however, to attain these rates of production, and the mining of lower grade ores would be needed to sustain them for more than a few years. By 1966 the U.S. economy will require at least 67 000 flasks, and the domestic demand for mercury thereafter will continue to increase. The probability that domestic mines will continue to supply a fourth or more of the domestic consumption is dependent upon the price of mercury for the next few years. The outlook for the long-term supply is reassuring, but it is dependent, not only upon price, but upon continued progress in new techniques of discovery.

Sensor Access to the Cellular Microenvironment Using the Sensing Cell Culture Flask.

PubMed

Kieninger, Jochen; Tamari, Yaara; Enderle, Barbara; Jobst, Gerhard; Sandvik, Joe A; Pettersen, Erik O; Urban, Gerald A

2018-04-26

The Sensing Cell Culture Flask (SCCF) is a cell culture monitoring system accessing the cellular microenvironment in 2D cell culture using electrochemical microsensors. The system is based on microfabricated sensor chips embedded in standard cell culture flasks. Ideally, the sensor chips could be equipped with any electrochemical sensor. Its transparency allows optical inspection of the cells during measurement. The surface of the sensor chip is in-plane with the flask surface allowing undisturbed cell growth on the sensor chip. A custom developed rack system allows easy usage of multiple flasks in parallel within an incubator. The presented data demonstrates the application of the SCCF with brain tumor (T98G) and breast cancer (T-47D) cells. Amperometric oxygen sensors were used to monitor cellular respiration with different incubation conditions. Cellular acidification was accessed with potentiometric pH sensors using electrodeposited iridium oxide films. The system itself provides the foundation for electrochemical monitoring systems in 3D cell culture.

Technical note: Method for isolation of the bovine sweat gland and conditions for in vitro culture.

PubMed

Hamzaoui, S; Burger, C A; Collier, J L; Collier, R J

2018-05-01

Apocrine sweat glands in bovine skin are involved in thermoregulation. Human, horse, and sheep sweat gland epithelial cells have been isolated and grown in vitro. The present study was conducted to identify a method to isolate bovine sweat glands and culture apocrine bovine sweat gland epithelial cells in vitro. Mechanical shearing, collagenase digestion, centrifugation, and neutral red staining were used to identify and isolate the apocrine glands from skin. Bovine sweat glands in situ and after isolation comprised 2 major cell types consisting of a single layer of cuboidal epithelial cells resting on a layer of myoepithelial cells. In situ, the glands were embedded in a collagen matrix primarily comprising fibroblasts, and some of these cells were also present in the isolated material. The isolated material was transferred to complete medium (keratinocyte serum-free medium, bovine pituitary extract, and human recombinant epidermal growth factor + 2.5% fetal bovine serum) in a T 25 flask (Falcon, Franklin Lakes, NJ) with media film and then incubated at 37°C for 24 h. After sweat glands adhered to the bottom of the flask, an additional 2 mL of complete medium was added and the medium was changed every 3 d. Isolated apocrine sweat glands and bovine sweat gland epithelial cells were immunostained for cytokeratin and fibroblast specific protein, indicating fibroblast-free cultures. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Influence of nutritional and physicochemical variables on PHB production from raw glycerol obtained from a Colombian biodiesel plant by a wild-type Bacillus megaterium strain.

PubMed

Moreno, Paalo; Yañez, Camilo; Cardozo, Nilo Sérgio Medeiros; Escalante, Humberto; Combariza, Marianny Y; Guzman, Carolina

2015-12-25

Biodegradable polymers are currently viable alternatives to traditional synthetic polymers. For instance, polyhydroxybutyrate (PHB) is intracellularly produced and accumulated by Bacillus species, among others. This study reports several wild-type Bacillus strains with the ability to accumulate PHB using raw glycerol from biodiesel production as the sole carbon source. Out of 15 strains from different sources, B. megaterium B2 was selected as the most promising strain for further statistical optimization of the medium composition. Plackett-Burman and central composite designs were used to establish key variables and optimal culture conditions for PHB production using both 250-mL shake flasks and a 7.5-L bioreactor. Temperature and concentrations of glycerol and Na2HPO4 are the experimental variables with the most significant influence on PHB production by B2. After 14 hours of fermentation in shake flasks with optimized medium, B2 produced 0.43 g/L of PHB with a 34% accumulation in the cells. In contrast, under the same conditions, a maximum PHB concentration of 1.20 g/L in the bioreactor was reached at 11 hours. These values correspond to a 48% and 314% increase in PHB production compared to the initial culture conditions. These results suggest the potential of B2 as a PHB producer using raw glycerol, which is an inexpensive, abundant and readily available carbon source. Copyright © 2015 Elsevier B.V. All rights reserved.

Defined Essential 8™ Medium and Vitronectin Efficiently Support Scalable Xeno-Free Expansion of Human Induced Pluripotent Stem Cells in Stirred Microcarrier Culture Systems

PubMed Central

Badenes, Sara M.; Fernandes, Tiago G.; Cordeiro, Cláudia S. M.; Boucher, Shayne; Kuninger, David; Vemuri, Mohan C.; Diogo, Maria Margarida; Cabral, Joaquim M. S.

2016-01-01

Human induced pluripotent stem (hiPS) cell culture using Essential 8™ xeno-free medium and the defined xeno-free matrix vitronectin was successfully implemented under adherent conditions. This matrix was able to support hiPS cell expansion either in coated plates or on polystyrene-coated microcarriers, while maintaining hiPS cell functionality and pluripotency. Importantly, scale-up of the microcarrier-based system was accomplished using a 50 mL spinner flask, under dynamic conditions. A three-level factorial design experiment was performed to identify optimal conditions in terms of a) initial cell density b) agitation speed, and c) to maximize cell yield in spinner flask cultures. A maximum cell yield of 3.5 is achieved by inoculating 55,000 cells/cm2 of microcarrier surface area and using 44 rpm, which generates a cell density of 1.4x106 cells/mL after 10 days of culture. After dynamic culture, hiPS cells maintained their typical morphology upon re-plating, exhibited pluripotency-associated marker expression as well as tri-lineage differentiation capability, which was verified by inducing their spontaneous differentiation through embryoid body formation, and subsequent downstream differentiation to specific lineages such as neural and cardiac fates was successfully accomplished. In conclusion, a scalable, robust and cost-effective xeno-free culture system was successfully developed and implemented for the scale-up production of hiPS cells. PMID:26999816

Optimization of Carbon and Nitrogen Sources for Extracellular Polymeric Substances Production by Chryseobacterium indologenes MUT.2

PubMed Central

Khani, Mojtaba; Bahrami, Ali; Chegeni, Asma; Ghafari, Mohammad Davoud; Mansouran Zadeh, ALi

2016-01-01

Background Bacterial Extracellular Polymeric Substances (EPS) are environmental friendly and versatile polymeric materials that are used in a wide range of industries such as: food, textile, cosmetics, and pharmaceuticals. To make the production process of the EPS cost-effective, improvements in the production yield is required which could be implemented through application of processes such as optimized culture conditions, and development of the strains with higher yield (e.g. through genetic manipulation), or using low-cost substrates. Objectives In this work, the effects of carbon and nitrogen sources were studied in order to improve the EPS production by the submerged cultivation of Chryseobacterium indologenes MUT.2. Materials and Methods The mesophilic microorganism Chryseobacterium indologenes MUT.2, was grown and maintained in the Luria Bertani agar. The initial basal medium contained: glucose (20 g.L-1), yeast extracts (5 g.L-1), K2HPO4 (6 g.L-1), NaH2PO4 (7 g.L-1), NH4CL (0.7 g.L-1), and MgSO4 (0.5 g.L-1). For evaluating the carbon and nitrogen sources’ effect on the fermentation performance, cultures were prepared in 500 mL flasks filled with 300 mL of the medium. The single-factor experiments based on statistics was employed to evaluate and optimize the carbon and nitrogen sources for EPS production in the liquid culture medium of Chryseobacterium indologenes MUT.2. Results The preferred carbon-sources, sucrose and glucose, commonly gave the highest EPS production of 8.32 and 6.37 g.L-1, respectively, and the maximum EPS production of 8.87 g.L-1 was achieved when glutamic acid (5 g.L-1) was employed as the nitrogen source. Conclusions In this work, the culture medium for production of EPS by Chryseobacterium indologenes MUT.2 was optimized. Compared to the basal culture medium in shake-flasks and stirred tank bioreactor, the use of optimized culture medium has resulted in a 53% and 73% increase in the EPS production, respectively. PMID:28959321

Design of an efficient medium for heterologous protein production in Yarrowia lipolytica: case of human interferon alpha 2b.

PubMed

Gasmi, Najla; Ayed, Atef; Nicaud, Jean-Marc; Kallel, Héla

2011-05-20

The non conventional yeast Yarrowia lipolytica has aroused a strong industrial interest for heterologous protein production. However most of the studies describing recombinant protein production by this yeast rely on the use of complex media, such media are not convenient for large scale production particularly for products intended for pharmaceutical applications. In addition medium composition can also affect the production yield. Hence it is necessary to design an efficient medium for therapeutic protein expression by this host. Five different media, including four minimal media and a complex medium, were assessed in shake flasks for the production of human interferon alpha 2b (hIFN α2b) by Y. lipolytica under the control of POX2 promoter inducible with oleic acid. The chemically defined medium SM4 formulated by Invitrogen for Pichia pastoris growth was the most suitable. Using statistical experimental design this medium was further optimized. The selected minimal medium consisting in SM4 supplemented with 10 mg/l FeCl₃, 1 g/l glutamate, 5 ml/l PTM1 (Pichia Trace Metals) solution and a vitamin solution composed of myo-inositol, thiamin and biotin was called GNY medium. Compared to shake flask, bioreactor culture in GNY medium resulted in 416-fold increase of hIFN α2b production and 2-fold increase of the biological activity. Furthermore, SM4 enrichment with 5 ml/l PTM1 solution contributed to protect hIFN α2b against the degradation by the 28 kDa protease identified by zymography gel in culture supernatant. The screening of the inhibitory effect of the trace elements present in PTM1 solution on the activity of this protease was achieved using a Box-Behnken design. Statistical data analysis showed that FeCl₃ and MnSO₄ had the most inhibitory effect. We have designed an efficient medium for large scale production of heterologous proteins by Y. lipolytica. The optimized medium GNY is suitable for the production of hIFN α2b with the advantage that no complex nitrogen sources with non-defined composition were required.

New use of broomcorn millets for production of granular cultures of aphid-pathogenic fungus Pandora neoaphidis for high sporulation potential and infectivity to Myzus persicae.

PubMed

Hua, Li; Feng, Ming-Guang

2003-10-24

Glutinous broomcorn millets from the crop Panicum miliaceum were first used as substrate to produce granular cultures of Pandora neoaphidis, an obligate fungal pathogen specific to aphids. Carrying a water content of 36.5% after being steamed in a regular autoclaving procedure, millet grains of each 15 g (dry weight) in a 100-ml flask were mixed with 3 ml modified Sabouraud dextrose broth containing half a mashed colony of P. neoaphidis grown on egg yolk milk agar and then incubated at 20 degrees C and a light/dark cycle of 12 h/12 h for 21 days. Based on individually monitoring conidial production potential of 20 millet grains sampled from an arbitrarily taken flask at 3-day intervals, the millet cultures incubated for 6-15 days were capable of producing 16.8-23.4 x 10(4) conidia per millet grain with conidial ejection lasting for up to 6 days. The cultured millet grains individually produced significantly more conidia than apterous adults of Myzus persicae killed by P. neoaphidis (8.4 x 10(4) conidia per cadaver) and sporulated twice longer. The modeling of time-dose-mortality data from bioassays on M. persicae apterae exposed to conidial showers from the cultured millet grains and the mycelial mats produced in liquid culture resulted in similar estimates of LC(50) (millets: 21.4, 7.3, and 4.9 conidia mm(-2) on days 5-7 after exposure; mycelial mats: 22.1, 10.6, and 7.7 conidia mm(-2)) although the LT(50) estimated at a given conidial concentration was slightly smaller for the millet cultures than for the mycelial mats. This indicates that the millet grains cultured with P. neoaphidis produced conidia as infective as or slightly more infective to M. persicae than those from the mycelial mats. Based on the sporulation potential, infectivity, and ease and cost of the millet cultures, the method developed in this study highly improved in vitro cultures of P. neoaphidis and may adapt to culturing other entomophthoralean fungi for microbial control of insect pests.

New Generation Flask Sampling Technology Final Report

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

Smith, James R.

Scientists are turning their focus to the Arctic, site of one of the strongest climate change signals. A new generation of technologies is required to function within that harsh environment, chart evolution of its trace gases and provide new kinds of information for models of the atmosphere. Our response to the solicitation tracks how global atmospheric monitoring was launched more than a half century ago; namely, acquisition of discrete samples of air by flask and subsequent analysis in the laboratory. AOS is proposing to develop a new generation of flask sampling technology. It will enable the new Arctic programs tomore » begin with objective high density sampling of the atmosphere by UAS. The Phase I program will build the prototype flask technology and show that it can acquire and store mol fractions of CH4 and CO2 and value of δ13C with good fidelity. A CAD model will be produced for the entire platform including a package with 100 flasks and the airframe with auto-pilot, electronic propulsion and ground-to-air communications. A mobile flask analysis station will be prototyped in Phase I and designed to final form in Phase II. It expends very small sample per analysis and will interface directly to the flask package integrated permanently into the UAS fuselage. Commercial Applications and Other Benefits: • The New Generation Flask Sampling Technology able to provide a hundred or more samples of air per UAS mission. • A mobile analysis station expending far less sample than the existing ones and small enough to be stationed at the remote sites of Arctic operations. • A new form of validation for continuous trace gas observations from all platforms including the small UAS. • Further demonstration to potential customers of the AOS capabilities to invent, build, deploy and exploit entire platforms for observations of Earth’s atmosphere and ocean. Key Words: Flask Sampler, Mobile Analysis Station, Trace Gas, CO2, CH4, δC13, UAS, Baseline Airborne Observatory, Arctic, Climate Change. Summary for Members of Congress: The air, land and sea of the Arctic combine to produce a large climate change signal. AOS is proposing to develop unmanned airborne technologies able to begin prompt, objective observations of the signal’s atmospheric component.« less

Efficient Extracellular Expression of Phospholipase D in Escherichia Coli with an Optimized Signal Peptide

NASA Astrophysics Data System (ADS)

Yang, Leyun; Xu, Yu; Chen, Yong; Ying, Hanjie

2018-01-01

New secretion vectors containing the synthetic signal sequence (OmpA’) was constructed for the secretory production of recombinant proteins in Escherichia coli. The E. coli Phospholipase D structural gene (Accession number:NC_018658) fused to various signal sequence were expressed from the Lac promoter in E. coli Rosetta strains by induction with 0.4mM IPTG at 28°C for 48h. SDS-PaGe analysis of expression and subcellular fractions of recombinant constructs revealed the translocation of Phospholipase D (PLD) not only to the medium but also remained in periplasm of E. coli with OmpA’ signal sequence at the N-terminus of PLD. Thus the study on the effects of various surfactants on PLD extracellular production in Escherichia coli in shake flasks revealed that optimal PLD extracellular production could be achieved by adding 0.4% Triton X-100 into the medium. The maximal extracellular PLD production and extracellular enzyme activity were 0.23mg ml-1 and 16U ml-1, respectively. These results demonstrate the possibility of efficient secretory production of recombinant PLD in E. coli should be a potential industrial applications.

Lead removal by Spirulina platensis biomass.

PubMed

Al-Homaidan, Ali A; Al-Abbad, Aljawharah F; Al-Hazzani, Amal A; Al-Ghanayem, Abdullah A; Alabdullatif, Jamila A

2016-01-01

In this investigation, we report on the biosorption of Pb (II) from aqueous solutions by the nonliving biomass of the micro-alga (cyanobacterium) Spirulina platensis. Propagation of the micro-alga was carried out in outside oblong raceway ponds. The biomass was cleaned, dried and used for the investigation. The effects of pH, adsorbent dose, temperature, initial concentration of Pb (II), and contact time on the adsorption of lead by the dry biomass were studied. The experiments were carried out in 250 ml conical flasks containing 100 ml of test solutions using an orbital incubator at 150 rpm. Concentrations of the metal before and after the experiments were measured using Atomic Absorption Spectrophotometer. Very high levels of Pb (II) removal (>91%) were obtained. The optimum conditions for maximal adsorption by S. platensis were found to be pH 3; 2 g of adsorbent dose; incubation at 26°C; 100 mg/l of lead initial concentration and 60 minutes of contact time. The experimental data fitted well with Freundlich isotherm equation with R(2) values greater than 0.97. Based on our results, we recommend the utilization of S. platensis biomass for heavy metal removal from aqueous solutions.

Bioprocess exploration for thermostable α-amylase production of a deep-sea thermophile Geobacillus sp. in high-temperature bioreactor.

PubMed

Jiang, Tao; Huang, Mengmeng; He, Hao; Lu, Jian; Zhou, Xiangshan; Cai, Menghao; Zhang, Yuanxing

2016-08-17

Geobacillus sp. 4j, a deep-sea high-salt thermophile, was found to produce thermostable α-amylase. In this work, culture medium and conditions were first optimized to enhance the production of thermostable α-amylase by statistical methodologies. The resulting extracellular production was increased by five times and reached 6.40 U/ml. Then, a high-temperature batch culture of the thermophile in a 15 l in-house-designed bioreactor was studied. The results showed that a relatively high dissolved oxygen (600 rpm and 15 l/min) and culture temperature of 60°C facilitated both cell growth and α-amylase production. Thus, an efficient fermentation process was established with initial medium of pH 6.0, culture temperature of 60°C, and dissolved oxygen above 20%. It gave an α-amylase production of 79 U/ml and productivity of 19804 U/l·hr, which were 10.8 and 208 times higher than those in shake flask, respectively. This work is useful for deep-sea high-salt thermophile culture, where efforts are lacking presently.

Scale-up from shake flasks to bioreactor, based on power input and Streptomyces lividans morphology, for the production of recombinant APA (45/47 kDa protein) from Mycobacterium tuberculosis.

PubMed

Gamboa-Suasnavart, Ramsés A; Marín-Palacio, Luz D; Martínez-Sotelo, José A; Espitia, Clara; Servín-González, Luis; Valdez-Cruz, Norma A; Trujillo-Roldán, Mauricio A

2013-08-01

Culture conditions in shake flasks affect filamentous Streptomyces lividans morphology, as well the productivity and O-mannosylation of recombinant Ala-Pro-rich O-glycoprotein (known as the 45/47 kDa or APA antigen) from Mycobacterium tuberculosis. In order to scale up from previous reported shake flasks to bioreactor, data from the literature on the effect of agitation on morphology of Streptomyces strains were used to obtain gassed volumetric power input values that can be used to obtain a morphology of S. lividans in bioreactor similar to the morphology previously reported in coiled/baffled shake flasks by our group. Morphology of S. lividans was successfully scaled-up, obtaining similar mycelial sizes in both scales with diameters of 0.21 ± 0.09 mm in baffled and coiled shake flasks, and 0.15 ± 0.01 mm in the bioreactor. Moreover, the specific growth rate was successfully scaled up (0.09 ± 0.02 and 0.12 ± 0.01 h(-1), for bioreactors and flasks, respectively), and the recombinant protein productivity measured by densitometry, as well. More interestingly, the quality of the recombinant glycoprotein measured as the amount of mannoses attached to the C-terminal of APA was also scaled- up; with up to five mannose residues in cultures carried out in shake flasks; and six in the bioreactor. However, final biomass concentration was not similar, indicating that although the process can be scaled-up using the power input, others factors like oxygen transfer rate, tip speed or energy dissipation/circulation function can be an influence on bacterial metabolism.

Modeling of growth and laccase production by Pycnoporus sanguineus.

PubMed

Saat, Muhammad Naziz; Annuar, Mohamad Suffian Mohamad; Alias, Zazali; Chuan, Ling Tau; Chisti, Yusuf

2014-05-01

Production of extracellular laccase by the white-rot fungus Pycnoporus sanguineus was examined in batch submerged cultures in shake flasks, baffled shake flasks and a stirred tank bioreactor. The biomass growth in the various culture systems closely followed a logistic growth model. The production of laccase followed a Luedeking-Piret model. A modified Luedeking-Piret model incorporating logistic growth effectively described the consumption of glucose. Biomass productivity, enzyme productivity and substrate consumption were enhanced in baffled shake flasks relative to the cases for the conventional shake flasks. This was associated with improved oxygen transfer in the presence of the baffles. The best results were obtained in the stirred tank bioreactor. At 28 °C, pH 4.5, an agitation speed of 600 rpm and a dissolved oxygen concentration of ~25 % of air saturation, the laccase productivity in the bioreactor exceeded 19 U L(-1 )days(-1), or 1.5-fold better than the best case for the baffled shake flask. The final concentration of the enzyme was about 325 U L(-1).

Enhanced solid-state citric acid bio-production using apple pomace waste through surface response methodology.

PubMed

Dhillon, G S; Brar, S K; Verma, M; Tyagi, R D

2011-04-01

  To evaluate the potential of apple pomace (AP) supplemented with rice husk for hyper citric acid production through solid-state fermentation by Aspergillus niger NRRL-567. Optimization of two key parameters, such as moisture content and inducer (ethanol and methanol) concentration was carried out by response surface methodology.   In this study, the effect of two crucial process parameters for solid-state citric acid fermentation by A. niger using AP waste supplemented with rice husk were thoroughly investigated in Erlenmeyer flasks through response surface methodology. Moisture and methanol had significant positive effect on citric acid production by A. niger grown on AP (P < 0·05). Higher values of citric acid on AP by A. niger (342·41gkg(-1) and 248·42gkg(-1) dry substrate) were obtained with 75% (v/w) moisture along with two inducers [3% (v/w) methanol and 3% (v/w) ethanol] with fermentation efficiency of 93·90% and 66·42%, respectively depending upon the total carbon utilized after 144h of incubation period. With the same optimized parameters, conventional tray fermentation was conducted. The citric acid concentration of 187·96gkg(-1) dry substrate with 3% (v/w) ethanol and 303·34gkg(-1) dry substrate with 3% (v/w) methanol were achieved representing fermentation efficiency of 50·80% and 82·89% in tray fermentation depending upon carbon utilization after 120h of incubation period.   Apple pomace proved to be the promising substrate for the hyper production of citric acid through solid-state tray fermentation, which is an economical technique and does not require any sophisticated instrumentation.   The study established that the utilization of agro-industrial wastes have positive repercussions on the economy and will help to meet the increasing demands of citric acid and moreover will help to alleviate the environmental problems resulting from the disposal of agro-industrial wastes. © 2011 The Authors. Journal of Applied Microbiology © 2011 The Society for Applied Microbiology.

Very high cell density perfusion of CHO cells anchored in a non-woven matrix-based bioreactor.

PubMed

Zhang, Ye; Stobbe, Per; Silvander, Christian Orrego; Chotteau, Véronique

2015-11-10

Recombinant Chinese Hamster Ovary (CHO) cells producing IgG monoclonal antibody were cultivated in a novel perfusion culture system CellTank, integrating the bioreactor and the cell retention function. In this system, the cells were harbored in a non-woven polyester matrix perfused by the culture medium and immersed in a reservoir. Although adapted to suspension, the CHO cells stayed entrapped in the matrix. The cell-free medium was efficiently circulated from the reservoir into- and through the matrix by a centrifugal pump placed at the bottom of the bioreactor resulting in highly homogenous concentrations of the nutrients and metabolites in the whole system as confirmed by measurements from different sampling locations. A real-time biomass sensor using the dielectric properties of living cells was used to measure the cell density. The performances of the CellTank were studied in three perfusion runs. A very high cell density measured as 200 pF/cm (where 1 pF/cm is equivalent to 1 × 10(6)viable cells/mL) was achieved at a perfusion rate of 10 reactor volumes per day (RV/day) in the first run. In the second run, the effect of cell growth arrest by hypothermia at temperatures lowered gradually from 37 °C to 29 °C was studied during 13 days at cell densities above 100 pF/cm. Finally a production run was performed at high cell densities, where a temperature shift to 31 °C was applied at cell density 100 pF/cm during a production period of 14 days in minimized feeding conditions. The IgG concentrations were comparable in the matrix and in the harvest line in all the runs, indicating no retention of the product of interest. The cell specific productivity was comparable or higher than in Erlenmeyer flask batch culture. During the production run, the final harvested IgG production was 35 times higher in the CellTank compared to a repeated batch culture in the same vessel volume during the same time period. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

Flow field measurements in the cell culture unit

NASA Technical Reports Server (NTRS)

Walker, Stephen; Wilder, Mike; Dimanlig, Arsenio; Jagger, Justin; Searby, Nancy

2002-01-01

The cell culture unit (CCU) is being designed to support cell growth for long-duration life science experiments on the International Space Station (ISS). The CCU is a perfused loop system that provides a fluid environment for controlled cell growth experiments within cell specimen chambers (CSCs), and is intended to accommodate diverse cell specimen types. Many of the functional requirements depend on the fluid flow field within the CSC (e.g., feeding and gas management). A design goal of the CCU is to match, within experimental limits, all environmental conditions, other than the effects of gravity on the cells, whether the hardware is in microgravity ( micro g), normal Earth gravity, or up to 2g on the ISS centrifuge. In order to achieve this goal, two steps are being taken. The first step is to characterize the environmental conditions of current 1g cell biology experiments being performed in laboratories using ground-based hardware. The second step is to ensure that the design of the CCU allows the fluid flow conditions found in 1g to be replicated from microgravity up to 2g. The techniques that are being used to take these steps include flow visualization, particle image velocimetry (PIV), and computational fluid dynamics (CFD). Flow visualization using the injection of dye has been used to gain a global perspective of the characteristics of the CSC flow field. To characterize laboratory cell culture conditions, PIV is being used to determine the flow field parameters of cell suspension cultures grown in Erlenmeyer flasks on orbital shakers. These measured parameters will be compared to PIV measurements in the CSCs to ensure that the flow field that cells encounter in CSCs is within the bounds determined for typical laboratory experiments. Using CFD, a detailed simulation is being developed to predict the flow field within the CSC for a wide variety of flow conditions, including microgravity environments. Results from all these measurements and analyses of the CSC flow environment are presented and discussed. The final configuration of the CSC employs magnetic stir bars with angled paddles to achieve the necessary flow requirements within the CSC.

Delta 13C in CO2 at Alert, NWT, Canada (June 1991 - December 2001)

DOE Data Explorer

Allison, C. E. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Aspendale, Victoria, Australia; Francey, R. J. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Aspendale, Victoria, Australia; Krummel, P. B. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Aspendale, Victoria, Australia

2003-04-01

Measurements have been made on air collected in flasks at Alert, Canada, through the CSIRO GASLAB worldwide network. Flasks are filled with air at Alert and returned to the CSIRO GASLAB for analysis; typical sample storage times for flasks collected at Alert range from a few weeks to a few months. No significant effect on the stable carbon isotopic composition, δ13C, has been detected as a consequence of the sample storage time.

Formation of three-dimensional cell/polymer constructs for bone tissue engineering in a spinner flask and a rotating wall vessel bioreactor

NASA Technical Reports Server (NTRS)

Sikavitsas, Vassilios I.; Bancroft, Gregory N.; Mikos, Antonios G.; McIntire, L. V. (Principal Investigator)

2002-01-01

The aim of this study is to investigate the effect of the cell culture conditions of three-dimensional polymer scaffolds seeded with rat marrow stromal cells (MSCs) cultured in different bioreactors concerning the ability of these cells to proliferate, differentiate towards the osteoblastic lineage, and generate mineralized extracellular matrix. MSCs harvested from male Sprague-Dawley rats were culture expanded, seeded on three-dimensional porous 75:25 poly(D,L-lactic-co-glycolic acid) biodegradable scaffolds, and cultured for 21 days under static conditions or in two model bioreactors (a spinner flask and a rotating wall vessel) that enhance mixing of the media and provide better nutrient transport to the seeded cells. The spinner flask culture demonstrated a 60% enhanced proliferation at the end of the first week when compared to static culture. On day 14, all cell/polymer constructs exhibited their maximum alkaline phosphatase activity (AP). Cell/polymer constructs cultured in the spinner flask had 2.4 times higher AP activity than constructs cultured under static conditions on day 14. The total osteocalcin (OC) secretion in the spinner flask culture was 3.5 times higher than the static culture, with a peak OC secretion occurring on day 18. No considerable AP activity and OC secretion were detected in the rotating wall vessel culture throughout the 21-day culture period. The spinner flask culture had the highest calcium content at day 14. On day 21, the calcium deposition in the spinner flask culture was 6.6 times higher than the static cultured constructs and over 30 times higher than the rotating wall vessel culture. Histological sections showed concentration of cells and mineralization at the exterior of the foams at day 21. This phenomenon may arise from the potential existence of nutrient concentration gradients at the interior of the scaffolds. The better mixing provided in the spinner flask, external to the outer surface of the scaffolds, may explain the accelerated proliferation and differentiation of marrow stromal osteoblasts, and the localization of the enhanced mineralization on the external surface of the scaffolds. Copyright 2002 Wiley Periodicals, Inc.

Development of a method for efficient cost-effective screening of Aspergillus niger mutants having increased production of glucoamylase.

PubMed

Zhu, Xudong; Arman, Bessembayev; Chu, Ju; Wang, Yonghong; Zhuang, Yingping

2017-05-01

To develop an efficient cost-effective screening process to improve production of glucoamylase in Aspergillus niger. The cultivation of A. niger was achieved with well-dispersed morphology in 48-deep-well microtiter plates, which increased the throughput of the samples compared to traditional flask cultivation. There was a close negative correlation between glucoamylase and its pH of the fermentation broth. A novel high-throughput analysis method using Methyl Orange was developed. When compared to the conventional analysis method using 4-nitrophenyl α-D-glucopyranoside as substrate, a correlation coefficient of 0.96 by statistical analysis was obtained. Using this novel screening method, we acquired a strain with an activity of 2.2 × 10 3  U ml -1 , a 70% higher yield of glucoamylase than its parent strain.

Development of a simple and low cost microbioreactor for high-throughput bioprocessing.

PubMed

Rahman, Pattanathu K S M; Pasirayi, Godfrey; Auger, Vincent; Ali, Zulfiqur

2009-02-01

A simple microbioreactor for high-throughput bioprocessing made from low cost polymer polytetrafluoroethylene (PTFE) tubes with a working volume of 1.5 ml is described. We have developed a microfluidic system that handles a small population of cells of a model microorganism, Pseudomonas aeruginosa DS10-129. Under the conditions of the microbioreactor, the organism produced extracellular secondary metabolites by using nutrient broth modified with glycerol. Pyocyanins were isolated from the fermented medium as a metabolite of interest. Antibiotic properties of pyocyanin were effective against a number of microorganisms such as Staphylococcus aureus, S. epidermis, Bacillus subtilis, Micrococcus luteus and Saccharomyces cerevisiae. Batch fermentation of the model organism in the microbioreactor was compared to shake-flask and conventional bench fermenter methods. Results obtained from the microbioreactor compared favourably with the conventional processes.

Pilot-scale production and liquid formulation of Rhodotorula minuta, a potential biocontrol agent of mango anthracnose.

PubMed

Patiño-Vera, M; Jiménez, B; Balderas, K; Ortiz, M; Allende, R; Carrillo, A; Galindo, E

2005-01-01

To develop a pilot-plant fermentation process for the production of the yeast Rhodotorula minuta, to be used as a biocontrol agent of mango anthracnose, using a low-cost culture medium. To develop a stable liquid formulation that preserve high viability of the yeast stored at 4 degrees C. Keeping constant the volumetric power input, a fermentation process was scaled-up from shake flasks to a 100 l bioreactor. Preharvest applications of the yeast resulted in postharvest anthracnose severity equal or lower than that observed with a chemical fungicide. Glycerol was added to the formulation as water activity reducer and xanthan gum as a viscosity-enhancing agent. Yeast initial concentration of 10(10) CFU ml(-1) resulted in 4-5 orders of magnitude decrease after 1 month of storage at 4 degrees C, whereas when it was formulated at 10(9) CFU ml(-1), the decrease was of two orders of magnitude in 6 months. The fermentation process was successfully scaled-up using a low-cost culture medium. Postharvest anthracnose severity could be considerably reduced using this yeast. Formulating the yeast at 10(9) CFU ml(-1) and adding glycerol (20%) and xanthan (5 g l(-1)) avoided both contamination and yeast sedimentation and it was able to preserve up to 10(7) CFU ml(-1) after 6 months at 4 degrees C. The yeast R. minuta is reported as a novel antagonistic micro-organism against the pathogen Colletotrichum gloeosporioides. Pilot plant production of this yeast allowed us to conduct field tests in commercial orchards during three harvest seasons. Yeast suspensions applied to mango trees reduced the fruit anthracnose severity in levels similar or better than chemical fungicides.

Evaluation of calcium alginate beads for Ce, La and Nd preconcentration from groundwater prior to ICP OES analysis.

PubMed

Arantes de Carvalho, Gabriel G; Kondaveeti, Stalin; Petri, Denise F S; Fioroto, Alexandre M; Albuquerque, Luiza G R; Oliveira, Pedro V

2016-12-01

Analytical methods for the determination of rare earth elements (REE) in natural waters by plasma spectrochemical techniques often require sample preparation procedures for analytes preconcentration as well as for removing matrix constituents, that may interfere on the analytical measurements. In the present work, calcium alginate (CA) beads were used for the first time aiming at Ce, La and Nd preconcentration from groundwater samples for further determination by inductively coupled plasma optical emission spectrometry (ICP OES). Test samples were analyzed in batch mode by transferring a 40mL test portion (pH=5±0.2) into a 50mL polyethylene flask containing 125mg CA beads. After 15min contact, the analytes were quantitatively extracted from the loaded CA beads with 2.0mL of 1.0molL -1 HCl solution for further determination by ICP OES, using Ce (II) 456.236, La (II) 379.478 and Nd (II) 430.358nm emission lines. The proposed approach is a reliable alternative for REE single-stage preconcentration from aqueous samples, as it provided accurate results based on the addition and recovery analysis of groundwater. The results obtained by the proposed method were also compared with those from reference method based on inductively coupled plasma mass spectrometry (ICP-MS) and no significant differences were observed after applying the Student's t-test at 95% confidence level. Copyright © 2016 Elsevier B.V. All rights reserved.

21 CFR 172.250 - Petroleum naphtha.

Code of Federal Regulations, 2010 CFR

2010-04-01

... at room temperature, with intermittent swirling. At the end of this time, disconnect the flask and.... Dissolve the crystals in the flask with about 25 milliliters of distilled water and pour this also into the...

21 CFR 172.250 - Petroleum naphtha.

Code of Federal Regulations, 2011 CFR

2011-04-01

... at room temperature, with intermittent swirling. At the end of this time, disconnect the flask and.... Dissolve the crystals in the flask with about 25 milliliters of distilled water and pour this also into the...

21 CFR 172.250 - Petroleum naphtha.

Code of Federal Regulations, 2014 CFR

2014-04-01

.... At the end of this time, disconnect the flask and evaporate the methyl alcohol on the steam bath... transfer the isooctane extract to a 250-milliliter separatory funnel. Dissolve the crystals in the flask...

21 CFR 172.250 - Petroleum naphtha.

Code of Federal Regulations, 2012 CFR

2012-04-01

... at room temperature, with intermittent swirling. At the end of this time, disconnect the flask and.... Dissolve the crystals in the flask with about 25 milliliters of distilled water and pour this also into the...

21 CFR 172.250 - Petroleum naphtha.

Code of Federal Regulations, 2013 CFR

2013-04-01

... at room temperature, with intermittent swirling. At the end of this time, disconnect the flask and.... Dissolve the crystals in the flask with about 25 milliliters of distilled water and pour this also into the...

Atmospheric Hydrogen (H2) Concentrations from the CSIRO GASLAB Flask Sampling Network (1992 - 2001)

DOE Data Explorer

Steele, L. P. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Atmospheric Research, Aspendale, Victoria, Australia; Krummel, P. B. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Atmospheric Research, Aspendale, Victoria, Australia; Langenfelds, R. L. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Atmospheric Research, Aspendale, Victoria, Australia

2003-01-01

Air samples from nine sites were collected from the CSIRO GASLAB Flask Sampling Network for the purpose of monitoring the atmospheric hydrogen (H2) concentrations. The listed data were obtained from flask air samples returned to the CSIRO GASLAB for analysis. Typical sample storage times ranged from days to weeks for some sites (e.g., Cape Grim) to as much as one year for Macquarie Island and the Antarctic sites. Experiments carried out to test for any change in sample H22 mixing ratio during storage have shown no consistent and systematic drift in these flask types over test periods of several months to years (Cooper et al., 1999). An annual cycle of H2 is evident, reflecting the seasonal nature of some of the major sources and sinks (Novelli et al., 1999).

A comparison of the effects of two methods of acclimation of aerobic biodegradability

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

Watson, H.M.

1993-11-01

The acclimation or adaptation of microorganisms to organic chemicals is an important factor influencing both the rate and the extent of biodegradation. In this study two acclimation procedures were evaluated in terms of their effectiveness in enhancing biodegradation, their relative ease of use in the laboratory, and the implications for biodegradability testing. In the single-flask procedure, microorganisms were acclimated for 2 to 7 d in a single acclimation flask at constant or increasing concentrations of the test chemical without transfer of microorganisms. In the second procedure, the enrichment procedure, microorganisms were acclimated in a series of flasks over a 21-dmore » period by making adaptive transfers to increasing concentrations of the test chemical. Acclimated microorganisms from each procedure were used as the source of inoculum for subsequent biodegradation tests in which carbon dioxide evolution was measured. Six chemicals were tested: quinoline, p-nitrophenol, N-methylaniline, N,N-dimethylaniline, acrylonitrile, and 2,2,4-trimethyl-1,3-pentanediol monoisobutyrate. Microorganisms acclimated in the single-flask procedure were much more effective than those acclimated in the enrichment procedure in degrading the test chemicals. The single-flask procedure is more convenient to use, and it permits monitoring of the time needed for acclimation. The results from these studies have implications for the methodology used in biodegradation test systems and suggest caution before adopting a multiple-flask, enrichment acclimation procedure before the performance of standardized tests for aerobic biodegradability.« less

Sampling, storage, and analysis of C2-C7 non-methane hydrocarbons from the US National Oceanic and Atmospheric Administration Cooperative Air Sampling Network glass flasks.

PubMed

Pollmann, Jan; Helmig, Detlev; Hueber, Jacques; Plass-Dülmer, Christian; Tans, Pieter

2008-04-25

An analytical technique was developed to analyze light non-methane hydrocarbons (NMHC), including ethane, propane, iso-butane, n-butane, iso-pentane, n-pentane, n-hexane, isoprene, benzene and toluene from whole air samples collected in 2.5l-glass flasks used by the National Oceanic and Atmospheric Administration, Earth System Research Laboratory, Global Monitoring Division (NOAA ESRL GMD, Boulder, CO, USA) Cooperative Air Sampling Network. This method relies on utilizing the remaining air in these flasks (which is at below-ambient pressure at this stage) after the completion of all routine greenhouse gas measurements from these samples. NMHC in sample aliquots extracted from the flasks were preconcentrated with a custom-made, cryogen-free inlet system and analyzed by gas chromatography (GC) with flame ionization detection (FID). C2-C7 NMHC, depending on their ambient air mixing ratios, could be measured with accuracy and repeatability errors of generally < or =10-20%. Larger deviations were found for ethene and propene. Hexane was systematically overestimated due to a chromatographic co-elution problem. Saturated NMHC showed less than 5% changes in their mixing ratios in glass flask samples that were stored for up to 1 year. In the same experiment ethene and propene increased at approximately 30% yr(-1). A series of blank experiments showed negligible contamination from the sampling process and from storage (<10 pptv yr(-1)) of samples in these glass flasks. Results from flask NMHC analyses were compared to in-situ NMHC measurements at the Global Atmospheric Watch station in Hohenpeissenberg, Germany. This 9-months side-by-side comparison showed good agreement between both methods. More than 94% of all data comparisons for C2-C5 alkanes, isoprene, benzene and toluene fell within the combined accuracy and precision objectives of the World Meteorological Organization Global Atmosphere Watch (WMO-GAW) for NMHC measurements.

A mini-scale mass production and separation system for secretory heterologous proteins by perfusion culture of recombinant Pichia pastoris using a shaken ceramic membrane flask.

PubMed

Ohashi, R; Mochizuki, E; Suzuki, T

1999-01-01

The perfusion culture technique using a shaken ceramic membrane flask (SCM flask) was applied to the production of a secretory heterologous protein. A recombinant methylotrophic yeast strain, Pichia pastoris, was cultured aerobically on a reciprocal shaker using an SCM flask. High-level production of human serum albumin (HSA) was attempted by increasing both the cell concentration and the expression level of the recombinant gene. In the two-stage culture method, the cell concentration was first raised to 17 g/l by feeding glycerol, after which the expression of HSA was induced by feeding methanol. However, the concentration of HSA in the effluent filtrate was as low as 0.15 g/l, while the cell concentration continued to increase. In contrast, HSA was effectively produced by feeding methanol from an early stage of the culture. In this case, the HSA concentration reached 0.24 and 0.46 g/l, respectively, using the growth-associated production method without and with aeration into the head space of the SCM flask. The results showed that supplying sufficient oxygen together with the growth-associated induction method are effective for obtaining high-level expression of the methanol-inducible recombinant gene of P. pastoris. An HSA concentration in the filtrate of 1.5 g/l was finally achieved when the cell concentration was increased to 53 g/l by supplying oxygen-enriched gas to the SCM flask. The yield and productivity of HSA reached 2.6-fold and 10-fold those obtained in an ordinary fed-batch culture using a shake flask, and these levels were readily achieved by continuous replenishment of the culture supernatant. The achievements made in this study should contribute to the development of a handy bioreactor system for mini-scale mass production of target proteins with separation at high purity.

The Safe and Efficient Evaporation of a Solvent from Solution

NASA Astrophysics Data System (ADS)

Mahon, Andrew R.

1997-02-01

The process of evaporating a solvent from a solution can cause problems for many students. By using a water-vacuum aspirator, backflashes of water can flood the sample tube and be detrimental to the experiment. This type of apparatus can also cause problems by drawing the solution it is evaporating back into the vacuum hose, causing the student to lose part or all of the products of their experiment. Macroscale and Microscale Organic Experiments, 2nd edition (1), suggested two techniques to dissolve solvents from a mixture. It suggested blowing a stream of air over the solution from a Pasteur pipet, or attaching a Pasteur pipet to an aspirator and drawing air over the surface of the liquid. Again, the danger of blowing air over the solution leaves the risk of splattering the solution, and drawing air over the surface of the liquid as described further endangers the products of the experiment through the risk of sucking the products up into the pipet aspirator. In an effort to eliminate these problems, a new technique has been developed. By inverting an ordinary 200-mL vacuum flask and pulling a steady current of air from the vacuum apparatus through it, any type of small container can be placed under it, allowing the solvent to be evaporated in a steady, mistake-free manner . By evaporating the solvent from the container that the products will be submitted in, no sample is lost through the process of transferring it from a vacuum flask or beaker to the final container.

Liquid films on shake flask walls explain increasing maximum oxygen transfer capacities with elevating viscosity.

PubMed

Giese, Heiner; Azizan, Amizon; Kümmel, Anne; Liao, Anping; Peter, Cyril P; Fonseca, João A; Hermann, Robert; Duarte, Tiago M; Büchs, Jochen

2014-02-01

In biotechnological screening and production, oxygen supply is a crucial parameter. Even though oxygen transfer is well documented for viscous cultivations in stirred tanks, little is known about the gas/liquid oxygen transfer in shake flask cultures that become increasingly viscous during cultivation. Especially the oxygen transfer into the liquid film, adhering on the shake flask wall, has not yet been described for such cultivations. In this study, the oxygen transfer of chemical and microbial model experiments was measured and the suitability of the widely applied film theory of Higbie was studied. With numerical simulations of Fick's law of diffusion, it was demonstrated that Higbie's film theory does not apply for cultivations which occur at viscosities up to 10 mPa s. For the first time, it was experimentally shown that the maximum oxygen transfer capacity OTRmax increases in shake flasks when viscosity is increased from 1 to 10 mPa s, leading to an improved oxygen supply for microorganisms. Additionally, the OTRmax does not significantly undermatch the OTRmax at waterlike viscosities, even at elevated viscosities of up to 80 mPa s. In this range, a shake flask is a somehow self-regulating system with respect to oxygen supply. This is in contrary to stirred tanks, where the oxygen supply is steadily reduced to only 5% at 80 mPa s. Since, the liquid film formation at shake flask walls inherently promotes the oxygen supply at moderate and at elevated viscosities, these results have significant implications for scale-up. © 2013 Wiley Periodicals, Inc.

THE FEDERAL INTEGRATED BIOTREATMENT RESEARCH CONSORTIUM (FLASK TO FIELD)

EPA Science Inventory

The Federal Integrated Biotreatment Research Consortium (Flask to Field) represented a 7-year concerted effort by several research laboratories to develop bioremediation technologies for contaminated DoD sites. The consortium structure consisted of a director and four thrust are...

The Oxygen Flask Method

ERIC Educational Resources Information Center

Boulton, L. H.

1973-01-01

Discusses application of Schoniger's method of quantitative organic elemental analysis in teaching of qualitative analysis of the halogens, nitrogen, sulphur, and phosphorus. Indicates that the oxygen flask method is safe and suitable for both high school and college courses because of simple apparatus requirements. (CC)

Methods of Sensing Land Pollution from Sanitary Landfills

NASA Technical Reports Server (NTRS)

Nosanov, Myron Ellis; Bowerman, Frank R.

1971-01-01

Major cities are congested and large sites suitable for landfill development are limited. Methane and other gases are produced at most sanitary landfills and dumps. These gases may migrate horizontally and vertically and have caused fatalities. Monitoring these gases provides data bases for design and construction of safe buildings on and adjacent to landfills. Methods of monitoring include: (1) a portable combustible gas indicator; and (2) glass flasks valved to allow simultaneous exhaust of the flask and aspiration of the sample into the flask. Samples are drawn through tubing from probes as deep as twenty-five feet below the surface.

Conducting Miller-Urey Experiments

NASA Technical Reports Server (NTRS)

Parker, Eric Thomas; Cleaves, Henderson James; Burton, Aaron S.; Glavin, Daniel P.; Dworkin, Jason; Zhou, Manshui; Bada, Jeffrey L.; Fernandez, Facundo M.

2014-01-01

In 1953, Stanley Miller reported the production of biomolecules from simple gaseous starting materials, using apparatus constructed to simulate the primordial Earth's atmosphere-ocean system. Miller introduced 200 ml of water, 100 mmHg of H2, 200mmHg of CH4, and 200mmHg of NH3 into the apparatus, then subjected this mixture, under reflux, to an electric discharge for a week, while the water was simultaneously heated. The purpose of this manuscript is to provide the reader with a general experimental protocol that can be used to conduct a Miller-Urey type spark discharge experiment, using a simplified 3 L reaction flask. Since the experiment involves exposing inflammable gases to a high voltage discharge, it is worth highlighting important steps that reduce the risk of explosion. The general procedures described in this work can be extrapolated to design and conduct a wide variety of electric discharge experiments simulating primitive planetary environments.

Synthesis of Prebiotic Caramels Catalyzed by Ion-Exchange Resin Particles: Kinetic Model for the Formation of Di-d-fructose Dianhydrides.

PubMed

Ortiz Cerda, Imelda-Elizabeth; Thammavong, Phahath; Caqueret, Vincent; Porte, Catherine; Mabille, Isabelle; Garcia Fernandez, José Manuel; Moscosa Santillan, Mario; Havet, Jean-Louis

2018-02-21

Caramel enriched in di-d-fructose dianhydrides (DFAs, a family of prebiotic cyclic fructodisaccharides) is a functional food with beneficial properties for health. The aim of this work was to study the conversion of fructose into DFAs catalyzed by acid ion-exchange resin, in order to establish a simplified mechanism of the caramelization reaction and a kinetic model for DFA formation. Batch reactor experiments were carried out in a 250 mL spherical glass flask and afforded up to 50% DFA yields. The mechanism proposed entails order 2 reactions that describe fructose conversion on DFAs or formation of byproducts such as HMF or melanoidines. A third order 1 reaction defines DFA transformation into fructosyl-DFAs or fructo-oligosaccharides. The influence of fructose concentration, resin loading and temperature was studied to calculate the kinetic parameters necessary to scale up the process.

Downscaling the in vitro test of fungal bioremediation of polycyclic aromatic hydrocarbons: methodological approach.

PubMed

Drevinskas, Tomas; Mickienė, Rūta; Maruška, Audrius; Stankevičius, Mantas; Tiso, Nicola; Mikašauskaitė, Jurgita; Ragažinskienė, Ona; Levišauskas, Donatas; Bartkuvienė, Violeta; Snieškienė, Vilija; Stankevičienė, Antanina; Polcaro, Chiara; Galli, Emanuela; Donati, Enrica; Tekorius, Tomas; Kornyšova, Olga; Kaškonienė, Vilma

2016-02-01

The miniaturization and optimization of a white rot fungal bioremediation experiment is described in this paper. The optimized procedure allows determination of the degradation kinetics of anthracene. The miniaturized procedure requires only 2.5 ml of culture medium. The experiment is more precise, robust, and better controlled comparing it to classical tests in flasks. Using this technique, different parts, i.e., the culture medium, the fungi, and the cotton seal, can be analyzed. A simple sample preparation speeds up the analytical process. Experiments performed show degradation of anthracene up to approximately 60% by Irpex lacteus and up to approximately 40% by Pleurotus ostreatus in 25 days. Bioremediation of anthracene by the consortium of I. lacteus and P. ostreatus shows the biodegradation of anthracene up to approximately 56% in 23 days. At the end of the experiment, the surface tension of culture medium decreased comparing it to the blank, indicating generation of surfactant compounds.

Modeling and optimization of fermentation variables for enhanced production of lactase by isolated Bacillus subtilis strain VUVD001 using artificial neural networking and response surface methodology.

PubMed

Venkateswarulu, T C; Prabhakar, K Vidya; Kumar, R Bharath; Krupanidhi, S

2017-07-01

Modeling and optimization were performed to enhance production of lactase through submerged fermentation by Bacillus subtilis VUVD001 using artificial neural networks (ANN) and response surface methodology (RSM). The effect of process parameters namely temperature (°C), pH, and incubation time (h) and their combinational interactions on production was studied in shake flask culture by Box-Behnken design. The model was validated by conducting an experiment at optimized process variables which gave the maximum lactase activity of 91.32 U/ml. Compared to traditional activity, 3.48-folds improved production was obtained after RSM optimization. This study clearly shows that both RSM and ANN models provided desired predictions. However, compared with RSM (R 2  = 0.9496), the ANN model (R 2  = 0.99456) gave a better prediction for the production of lactase.

Enhancement in lipid content of Chlorella sp. MJ 11/11 from the spent medium of thermophilic biohydrogen production process.

PubMed

Ghosh, Supratim; Roy, Shantonu; Das, Debabrata

2017-01-01

The present study investigates the effect of spent media of acetogenic dark fermentation for mixotrophic algal cultivation for biodiesel production. Mixotrophic growth conditions were optimized in culture flask (250mL) using Chlorella sp. MJ 11/11. Maximum lipid accumulation (58% w/w) was observed under light intensity, pH, nitrate and phosphate concentration of 100μmolm -2 s -1 , 7, 2.7mM and 1.8mM, respectively. Air lift (1.4L) and flat panel (1.4L) reactors were considered for algal cultivation. Air lift showed significant improvement in biomass and lipid production as compared to flat panel reactor. The results could help in development of sustainable technology involving acetogenic hydrogen production integrated with sequential mitigation of spent media by algal cultivation for improved energy recovery. Copyright © 2016 Elsevier Ltd. All rights reserved.

Isolation and identification of a novel Candida sp. H2 producing D-arabitol and optimization of D-arabitol production.

PubMed

Song, Weibin; Lin, Yanqing; Hu, Haiyan; Xie, Zhipeng; Zhang, Jianguo

2011-03-01

To isolate a new osmophilic yeast for producing D-arabitol and research its optimal fermentation conditions for highest yield of D-arabitol from glucose. The isolated strain was characterized by electron microscopy, Biolog (GN) test, G + C content measurement and 26S rDNA D1/D2 domain sequences analysis. The purified fermentation product was identified by IR, 1H-NMR, 13C-NMR, MS and optical rotation analysis. Then the fermentation conditions for D-arabitol production were optimized. A new osmophilic yeast was isolated and identified as Candida sp. H2. Through the single factor experiment,the optimum conditions of 250 g/L glucose,10 g/L yeast extract, initial pH 6.0, 35 degrees C of culture temperature, 200 r/min of agitation, 200 mL medium in a 1000 mL flask of broth content, 1% (v/v) of inoculum size, 96 h of fermentation time were achieved. Based on the conditions above,weight yield of 35% (86.55 g D-arabitol from 250 g glucose) was obtained and 10% higher than the conditions not optimized. Candida sp. H2 was a novel strain for producing D-arabitol and valuable for further study.

[Isolation, identification and fermentation optimization of Bacillus tequilensis PanD37 producing L-aspartate α- decarboxylase].

PubMed

Feng, Zhibin; Zhang, Juan; Chen, Guozhong; Cha, Yaping; Liu, Jinjie; Ge, Yihe; Cheng, Shiwei; Yu, Botao

2016-01-04

We screened bacteria producing L-aspartate α-decarboxylase from grapery soil and optimized the fermentation conditions. L-aspartate α-decarboxylase producing bacteria were screened by color-changing circle and liquid secondary screening culture media. Combination of morphological, physiological and biochemical characteristics and 16S rRNA sequence analysis were used to identify the bacteria. Fermentation conditions were optimized by single factor test and orthogonal experiment. Strain PanD37 showed high L-aspartate α-decarboxylase producing property and was identified as Bacillus tequilensis. The optimum fermentation conditions of PanD37 were liquid volume of 50 mL in 500 mL flask, 220 r/min at 35 °C, inoculation amount of 5% for 28 h with a medium of 22.5 g/L sucrose, 7.5 g/L fumaric acid, 20 g/L peptone, 6 g/L L-aspartic acid, 2 g/L Triton X-100, at initial pH of 7.0. Under the optimal fermentation conditions, the highest L-aspartate α-decarboxylase activity reached 44.57 U/mL, which was 2.57 folds higher than that obtained before optimization. Strain PanD37 was identified as Bacillus tequilensiswhich was capable of highly producing L-aspartate α-decarboxylase under the optimal fermentation conditions.

Ethanol from Sugar Cane: Flask Experiments Using the EX-FERM Technique

PubMed Central

Rolz, Carlos; de Cabrera, Sheryl

1980-01-01

Alcohol production at the laboratory scale from sugar cane pieces by the EX-FERM technique was studied with 37 strains of Saccharomyces spp. The EX-FERM process is novel in that it employs the simultaneous extraction and fermentation of the sucrose in a cane-water suspension. Two types of cane treatments were used: chips and shredded pith, either fresh or dried. A mother culture of the yeast was prepared in enriched cane juice and then added to the cane-water mixture. After static fermentation for 40 h at 30°C, the cane was removed, and fresh cane was added to the yeast-alcohol broth. After an additional 24 h, the cane was again removed and the liquor was analyzed. After the first 40-h cycle, sugar consumption was above 99% with 10 of the 37 yeast strains tested, and ethanol reached levels of 1.29 to 4.00 g per 100 ml, depending on the yeast strain. The final ethanol concentration reached 4.27 to 5.37 g per 100 ml, and sugar consumption was above 98% in three cases during a second EX-FERM cycle employing previously air-dried chips and pith. Product yields were within accepted values. Cane treatment did not appear to affect the results at this level. PMID:16345626

Optimization of the procedure for counting the eggs of Fasciola gigantica in bovine faeces.

PubMed

Suhardono; Roberts, J A; Copeman, D B

2006-07-01

This paper describes a method for counting eggs of F. gigantica in bovine faeces that optimizes the proportion of eggs recovered and the repeatability of estimates. The method uses 3 g of faeces suspended in 0.05% Tween 20. The suspension is passed through three 6 cm diameter sieves in tandem to remove fibrous debris, with respective apertures of 1 mm, 450 microm, and either 266 or 200 microm. The filtrate is allowed to sediment for 3 min in a conical flask; the sediment is recovered, then resuspended in 200 ml of 0.05% Tween 20 and allowed to sediment. After 3 min the sediment is washed in a sieve with an aperture of 53 microm, which retains the eggs. Eggs suspended in 15 ml of 1% methylene blue are counted using a dissecting microscope. Use of Tween 20 instead of water as the suspending agent for faeces gave a significant threefold increased the proportion of eggs recovered and reduced variability between repeated counts. This method is able to detect about one-third of the eggs present. It was concluded that the high proportion of F. gigantica eggs lost may be due to the presence of hydrophobic and covalent bonds on the eggs that bind them to debris, with which they are discarded.

[Determination of olive oil content in olive blend oil by headspace gas chromatography-mass spectrometry].

PubMed

Jiang, Wanfeng; Zhang, Ning; Zhang, Fengyan; Yang, Zhao

2017-07-08

A method for the determination of the content of olive oil in olive blend oil by headspace gas chromatography-mass spectrometry (SH-GC/MS) was established. The amount of the sample, the heating temperature, the heating time, the amount of injection, the injection mode and the chromatographic column were optimized. The characteristic compounds of olive oil were found by chemometric method. A sample of 1.0 g was placed in a 20 mL headspace flask, and heated at 180℃ for 2700 s. Then, 1.0 mL headspace gas was taken into the instrument. An HP-88 chromatographic column was used for the separation and the analysis was performed by GC/MS. The results showed that the linear range was 0-100%(olive oil content). The linear correlation coefficient ( r 2 ) was more than 0.995, and the limits of detection were 1.26%-2.13%. The deviations of olive oil contents in the olive blend oil were from -0.65% to 1.02%, with the relative deviations from -1.3% to 6.8% and the relative standard deviations from 1.18% to 4.26% ( n =6). The method is simple, rapid, environment friendly, sensitive and accurate. It is suitable for the determination of the content of olive oil in olive blend oil.

A Simple Method to Reduce both Lactic Acid and Ammonium Production in Industrial Animal Cell Culture

PubMed Central

Freund, Nathaniel W.; Croughan, Matthew S.

2018-01-01

Fed-batch animal cell culture is the most common method for commercial production of recombinant proteins. However, higher cell densities in these platforms are still limited due to factors such as excessive ammonium production, lactic acid production, nutrient limitation, and/or hyperosmotic stress related to nutrient feeds and base additions to control pH. To partly overcome these factors, we investigated a simple method to reduce both ammonium and lactic acid production—termed Lactate Supplementation and Adaptation (LSA) technology—through the use of CHO cells adapted to a lactate-supplemented medium. Using this simple method, we achieved a reduction of nearly 100% in lactic acid production with a simultaneous 50% reduction in ammonium production in batch shaker flasks cultures. In subsequent fed-batch bioreactor cultures, lactic acid production and base addition were both reduced eight-fold. Viable cell densities of 35 million cells per mL and integral viable cell days of 273 million cell-days per mL were achieved, both among the highest currently reported for a fed-batch animal cell culture. Investigating the benefits of LSA technology in animal cell culture is worthy of further consideration and may lead to process conditions more favorable for advanced industrial applications. PMID:29382079

A Simple Method to Reduce both Lactic Acid and Ammonium Production in Industrial Animal Cell Culture.

PubMed

Freund, Nathaniel W; Croughan, Matthew S

2018-01-28

Fed-batch animal cell culture is the most common method for commercial production of recombinant proteins. However, higher cell densities in these platforms are still limited due to factors such as excessive ammonium production, lactic acid production, nutrient limitation, and/or hyperosmotic stress related to nutrient feeds and base additions to control pH. To partly overcome these factors, we investigated a simple method to reduce both ammonium and lactic acid production-termed Lactate Supplementation and Adaptation (LSA) technology-through the use of CHO cells adapted to a lactate-supplemented medium. Using this simple method, we achieved a reduction of nearly 100% in lactic acid production with a simultaneous 50% reduction in ammonium production in batch shaker flasks cultures. In subsequent fed-batch bioreactor cultures, lactic acid production and base addition were both reduced eight-fold. Viable cell densities of 35 million cells per mL and integral viable cell days of 273 million cell-days per mL were achieved, both among the highest currently reported for a fed-batch animal cell culture. Investigating the benefits of LSA technology in animal cell culture is worthy of further consideration and may lead to process conditions more favorable for advanced industrial applications.

Dispersant Effectiveness Of Heavy Fuel Oils Using The Baffled Flask Test

EPA Science Inventory

Dispersants have been widely used as a primary response measure for marine oil spills around the world. Recently, the U.S. Environmental Protection Agency (EPA) developed an improved laboratory dispersant testing protocol, called the Baffled Flask Test (BFT). The BFT protocol w...

Renewable Molecular Flasks with NADH Models: Combination of Light-Driven Proton Reduction and Biomimetic Hydrogenation of Benzoxazinones.

PubMed

Zhao, Liang; Wei, Jianwei; Lu, Junhua; He, Cheng; Duan, Chunying

2017-07-17

Using small molecules with defined pockets to catalyze chemical transformations resulted in attractive catalytic syntheses that echo the remarkable properties of enzymes. By modulating the active site of a nicotinamide adenine dinucleotide (NADH) model in a redox-active molecular flask, we combined biomimetic hydrogenation with in situ regeneration of the active site in a one-pot transformation using light as a clean energy source. This molecular flask facilitates the encapsulation of benzoxazinones for biomimetic hydrogenation of the substrates within the inner space of the flask using the active sites of the NADH models. The redox-active metal centers provide an active hydrogen source by light-driven proton reduction outside the pocket, allowing the in situ regeneration of the NADH models under irradiation. This new synthetic platform, which offers control over the location of the redox events, provides a regenerating system that exhibits high selectivity and efficiency and is extendable to benzoxazinone and quinoxalinone systems. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Corning HYPERFlask® for viral amplification and production of diagnostic reagents.

PubMed

Kearney, Brian J; Voorhees, Matthew A; Williams, Priscilla L; Olschner, Scott P; Rossi, Cynthia A; Schoepp, Randal J

2017-04-01

Viral preparations are essential components in diagnostic research and development. The production of large quantities of virus traditionally is done by infecting numerous tissue culture flasks or roller bottles, which require large incubators and/or roller bottle racks. The Corning HYPERFlask ® is a multilayer flask that uses a gas permeable film to provide gas exchange between the cells and culture medium and the atmospheric environment. This study evaluated the suitability of the HYPERFlask for production of Lassa, Ebola, Bundibugyo, Reston, and Marburg viruses and compared it to more traditional methods using tissue culture flasks and roller bottles. The HYPERFlask produced cultures were equivalent in virus titer and indistinguishable in immunodiagnostic assays. The use of the Corning HYPERFlask for viral production is a viable alternative to traditional tissue culture flasks and roller bottles. HYPERFlasks allow for large volumes of virus to be produced in a small space without specialized equipment. Copyright © 2016. Published by Elsevier B.V.

Strategies for Enhancing the Accumulation and Retention of Extracellular Matrix in Tissue-Engineered Cartilage Cultured in Bioreactors

PubMed Central

Shahin, Kifah; Doran, Pauline M.

2011-01-01

Production of tissue-engineered cartilage involves the synthesis and accumulation of key constituents such as glycosaminoglycan (GAG) and collagen type II to form insoluble extracellular matrix (ECM). During cartilage culture, macromolecular components are released from nascent tissues into the medium, representing a significant waste of biosynthetic resources. This work was aimed at developing strategies for improving ECM retention in cartilage constructs and thus the quality of engineered tissues produced in bioreactors. Human chondrocytes seeded into polyglycolic acid (PGA) scaffolds were cultured in perfusion bioreactors for up to 5 weeks. Analysis of the size and integrity of proteoglycans in the constructs and medium showed that full-sized aggrecan was being stripped from the tissues without proteolytic degradation. Application of low (0.075 mL min−1) and gradually increasing (0.075–0.2 mL min−1) medium flow rates in the bioreactor resulted in the generation of larger constructs, a 4.0–4.4-fold increase in the percentage of GAG retained in the ECM, and a 4.8–5.2-fold increase in GAG concentration in the tissues compared with operation at 0.2 mL min−1. GAG retention was also improved by pre-culturing seeded scaffolds in flasks for 5 days prior to bioreactor culture. In contrast, GAG retention in PGA scaffolds infused with alginate hydrogel did not vary significantly with medium flow rate or pre-culture treatment. This work demonstrates that substantial improvements in cartilage quality can be achieved using scaffold and bioreactor culture strategies that specifically target and improve ECM retention. PMID:21858004

Stress fermentation strategies for the production of hyperthermostable superoxide dismutase from Thermus thermophilus HB27: effects of ions.

PubMed

Zhu, Hu; Liu, Jianguo; Qu, Jianbo; Gao, Xinliang; Pan, Tao; Cui, Zhanfeng; Zhao, Xiubo; Lu, Jian R

2013-11-01

In this study, we explored how ammonium and metal ion stresses affected the production of recombinant hyperthermostable manganese superoxide dismutase (Mn-SOD). To improve Mn-SOD production, fed-batch culture in shake flasks and bioreactor fermentation were undertaken to examine the effects of [Formula: see text] and Mn(2+) feeding. Under the optimized feeding time and concentrations of [Formula: see text] and Mn(2+), the maximal SOD activity obtained from bioreactor fermentation reached some 480 U/ml, over 4 times higher than that in batch cultivation (113 U/ml), indicating a major enhancement of the concentration of Mn-SOD in the scale-up of hyperthermostable Mn-SOD production. In contrast, when the fed-batch culture with appropriate [Formula: see text] and Mn(2+) feeding was carried out in the same 5-L stirred tank bioreactor, a maximal SOD concentration of some 450 U/ml was obtained, again indicating substantial increase in SOD activity as a result of [Formula: see text] and Mn(2+) feeding. The isoelectric point (pI) of the sample was found to be 6.2. It was highly stable at 90 °C and circular dichroism measurements indicated a high α-helical content of 70 % as well, consistent with known SOD properties. This study indicates that [Formula: see text] and Mn(2+) play important roles in Mn-SOD expression. Stress fermentation strategies established in this study are useful for large-scale efficient production of hyperthermostable Mn-SOD and may also be valuable for the scale-up of other extremozymes.

Serum-free microcarrier based production of replication deficient Influenza vaccine candidate virus lacking NS1 using Vero cells

PubMed Central

2011-01-01

Background Influenza virus is a major health concern that has huge impacts on the human society, and vaccination remains as one of the most effective ways to mitigate this disease. Comparing the two types of commercially available Influenza vaccine, the live attenuated virus vaccine is more cross-reactive and easier to administer than the traditional inactivated vaccines. One promising live attenuated Influenza vaccine that has completed Phase I clinical trial is deltaFLU, a deletion mutant lacking the viral Nonstructural Protein 1 (NS1) gene. As a consequence of this gene deletion, this mutant virus can only propagate effectively in cells with a deficient interferon-mediated antiviral response. To demonstrate the manufacturability of this vaccine candidate, a batch bioreactor production process using adherent Vero cells on microcarriers in commercially available animal-component free, serum-free media is described. Results Five commercially available animal-component free, serum-free media (SFM) were evaluated for growth of Vero cells in agitated Cytodex 1 spinner flask microcarrier cultures. EX-CELL Vero SFM achieved the highest cell concentration of 2.6 × 10^6 cells/ml, whereas other SFM achieved about 1.2 × 10^6 cells/ml. Time points for infection between the late exponential and stationary phases of cell growth had no significant effect in the final virus titres. A virus yield of 7.6 Log10 TCID50/ml was achieved using trypsin concentration of 10 μg/ml and MOI of 0.001. The Influenza vaccine production process was scaled up to a 3 liter controlled stirred tank bioreactor to achieve a cell density of 2.7 × 10^6 cells/ml and virus titre of 8.3 Log10 TCID50/ml. Finally, the bioreactor system was tested for the production of the corresponding wild type H1N1 Influenza virus, which is conventionally used in the production of inactivated vaccine. High virus titres of up to 10 Log10 TCID50/ml were achieved. Conclusions We describe for the first time the production of Influenza viruses using Vero cells in commercially available animal-component free, serum-free medium. This work can be used as a basis for efficient production of attenuated as well as wild type Influenza virus for research and vaccine production. PMID:21835017

Preliminary report on part of the Oat Hill quicksilver mine, Mayacmas district, Napa County, California

USGS Publications Warehouse

Fix, Philip Forsyth

1955-01-01

Oat Hill quicksilver mine, located in the Mayacmas district of northern California, and credited with having produced more than 160,000 flasks of quicksilver, was sampled cooperatively by the Buray of Mines and Geological Survey during 1944. 28 diamond drill holes totaling 8,120 feet were drilled by the Bureau of Mines in four of the six principal veins to sample virgin low-grade reserves and stope fill, and reserves in the other two veins were estimated from existing underground workings and by inferences from drill holes in nearby veins. The writer estimates a total of 10,220 flasks of quicksilver in indicated and inferred reserves totaling 320,000 tons. Indicated reserves minable under 1943 conditions are estimated at 1,960 flasks of quicksilver in 75,000 tons averaging 3.0 lbs Hg per ton. Inferred reserves minable under 1943 conditions are estimated at 4,640 flasks of quicksilver in 109,920 tons averaging about 3.2 lbs Hg per ton. Inferred reserves believed minable only under economic conditions much more favorable than even those of 1943 are estimated at 2,620 flasks of quicksilver in 135,080 tons averaging a little less than 1.5 lbs Hg per ton. About two-thirds of the indicated reserves are accessible in underground workings. All other reserves are estimated approximately without access underground. Several areas not sampled may possibly contain reserves.

Geochemistry of selected mercury mine-tailings in the Parkfield Mercury District, California

USGS Publications Warehouse

Rytuba, James J.; Kotlyar, Boris B.; Wilkerson, Gregg; Olson, Jerry

2001-01-01

The Parkfield mercury district is located in the southern part of the California Coast Range mercury mineral belt and contains three silica-carbonate-type mercury deposits that have had significant mercury production. Mercury was first produced in the district in 1873, but the main period of production occurred from 1915-1922. Total production from the district is about 5,000 flasks of mercury (a flask equals 76 pounds of mercury) with most production coming from the Patriquin mine (1,875 flasks), and somewhat less from the King (1,600 flasks) and Dawson (1,470 flasks) mines. Several other small prospects and mines occur in the district but only minor production has come from them. In 1969, Phelan Sulphur Company carried out mineral exploration at the King mine and announced the discovery of 55,000 tons of mercury ore with an average grade of 5.2 pounds per ton. The King mine is located on federal land administered by the U.S. Bureau of Land Management. Several other parcels of federal land are present adjacent to other mines and prospects in the Parkfield district. An environmental assessment of mine sites on and adjacent to federal land was carried out to determine the amount of mercury and other trace metals present in mine wastes and in sediments from streams impacted by past mining.

On-line prediction of the glucose concentration of CHO cell cultivations by NIR and Raman spectroscopy: Comparative scalability test with a shake flask model system.

PubMed

Kozma, Bence; Hirsch, Edit; Gergely, Szilveszter; Párta, László; Pataki, Hajnalka; Salgó, András

2017-10-25

In this study, near-infrared (NIR) and Raman spectroscopy were compared in parallel to predict the glucose concentration of Chinese hamster ovary cell cultivations. A shake flask model system was used to quickly generate spectra similar to bioreactor cultivations therefore accelerating the development of a working model prior to actual cultivations. Automated variable selection and several pre-processing methods were tested iteratively during model development using spectra from six shake flask cultivations. The target was to achieve the lowest error of prediction for the glucose concentration in two independent shake flasks. The best model was then used to test the scalability of the two techniques by predicting spectra of a 10l and a 100l scale bioreactor cultivation. The NIR spectroscopy based model could follow the trend of the glucose concentration but it was not sufficiently accurate for bioreactor monitoring. On the other hand, the Raman spectroscopy based model predicted the concentration of glucose in both cultivation scales sufficiently accurately with an error around 4mM (0.72g/l), that is satisfactory for the on-line bioreactor monitoring purposes of the biopharma industry. Therefore, the shake flask model system was proven to be suitable for scalable spectroscopic model development. Copyright © 2017 Elsevier B.V. All rights reserved.

Resource estimation from historical data: Mercury, a test case

USGS Publications Warehouse

Cargill, S.M.; Root, D.H.; Bailey, E.H.

1980-01-01

A simple technique based on historical records of tonnage and grade of ore produced provides a means for calculating how much of a mineral product will be available in the future at various average grades. Estimates made on this basis are independent of geologic considerations or changing economic and political factors, although they are based on mining history, which was largely determined by these factors. The relatively minor element, mercury, was used for the test case reported here, but the method has been found applicable to forecasts of resources for other mineral products. Mercury resources available in ore in which the average grade is as low as 0.1% are estimated to be 53 ??106kg (1.5 ??106flasks) for the United States and 1551 ??106kg (45 ??106flasks) for the world; this amount is more than adequate to meet predicted demand to the year 2000. The expectable price of mercury in 1978 dollars at this 0.1% grade is projected to be $58.75 per kg ($2,025 per flask), but at a 10% annual inflation rate, it would be more than $12,000 per flask. To satisfy just the projected U.S. demand for mercury by 2000, the price is calculated to be $48.96 per kg ($1,688 per flask) in 1978 dollars at an average annual grade of 0.12%. ?? 1980 Plenum Publishing Corporation.

THE BAFFLED FLASK TEST FOR DISPERSANT EFFECTIVENESS: A ROUND ROBIN EVALUATION OF REPRODUCIBILITY AND REPEATABILITY

EPA Science Inventory

After two previous investigations demonstrated that the Baffled Flask Test (BFT) was an effective and reproducible method for screening the effectiveness of dispersant products in the laboratory, the USEPA decided that before the new protocol cold be considered for replacement of...

EVALUATION OF MIXING ENERGY IN LABORATORY FLASKS USED FOR DISPERSANT EFFECTIVENESS TESTING

EPA Science Inventory

The evaluation of dispersant effectiveness used for oil spills is commonly done using tests conducted in laboratory flasks. The success of a test relies on replication of the conditions at sea. We used a hot wire anemometer to characterize the turbulence characteristics in the s...

Fluid Thinking.

ERIC Educational Resources Information Center

Clift, Philip A.

1992-01-01

Provides a student worksheet and set-up instructions for a science experiment to demonstrate the concept of density. Students are asked to explain the phenomenon in which a liquid of lower density placed in the bottom of two vertical flasks flows up to replace the liquid of higher density in the upper flask. (MDH)

[Effects of naloxone on the expression of stem cell factor and C-kit receptor in combined oxygen-glucose deprivation of primary cultured human embryonic neuron in vitro].

PubMed

Zhu, Bo; Li, Lan-ying; Lü, Guo-yi; Xue, Yu-liang; Ye, Tie-hu

2010-04-01

To explore the effects of naloxone on the expression of c-kit receptor (c-kit R) and its ligand stem cell factor (SCF) in human embryo neuronal hypoxic injury. Serum-free cerebral cortical cultures prepared from embryonic human brains were deprived of both oxygen and glucose which would set up an environment more likely with that of in vivo ischemic injury. Neurons in 24-well culture plates were randomly divided into four groups: control group, hypoxia group, naloxone 0.5 microg/ml group and naloxone 10 microg/ml group. MTT assay and biological analysis were performed to study the cell death and the changes of extracellular concentrations of lactate dehydrogenase (LDH) after combined oxygen-glucose deprivation. Neurons in 25 ml culture flasks were also randomly allocated into four groups as previously described. Intracellular total RNA were extracted at different time points: pre-hypoxia, immediately after hypoxia, and 3, 6, 12, and 24 hours after reoxygenation. The changes of SCF/c-kit R mRNA expression in hypoxic neurons treated with different concentrations of naloxone pre and post oxygen-glucose deprivation were determined with RT-PCR. The cell vitality detected by MTT assay decreased significantly in hypoxia group and naloxone 0.5 microg/ml group when compared with control group (P<0.01), while no significant difference was found between naloxone 0.5 microg/ml group and hypoxia group or between naloxone 10 microg/ml group and control group. Extracellular concentration of LDH significantly increased in hypoxia group (P<0.05), while no difference was found between naloxone 0.5 microg/ml group and control group, between naloxone 0.5 microg/ml and hypoxia group, or between naloxone 10 microg/ml and control group (all P>0.05). Immediately after oxygen-glucose deprivation, the expression of SCF/c-kit R mRNA increased significantly (P<0.01). Among those the expression of SCF presented a distribution of double-peak value within 24 hours. After treated with different concentrations of naloxone, the peak value of each group were delayed to appear and went down with the increasing of naloxone concentration. The peak values in all treated groups were significantly different from that in control group (P<0.01). The expression of SCF/c-kit R mRNA increases at the early stage after combined oxygen-glucose deprivation. Naloxone 0.5 microg/ml can attenuate cell injuries and regulate the expression of SCF/c-kit R. Naloxone may protect neurons by modulating the expressions of some cytokines.

Condensation of acetol and acetic acid vapor with sprayed liquid

USDA-ARS?s Scientific Manuscript database

A cellulose-derived fraction of biomass pyrolysis vapor was simulated by evaporating acetol and acetic acid (AA) from flasks on a hot plate. The liquid in the flasks was infused with heated nitrogen. The vapor/nitrogen stream was superheated in a tube oven and condensed by contact with a cloud of ...

PHYSICAL AND BIOLOGICAL PARAMETERS THAT DETERMINE THE FATE OF 'P'-CHLOROPHENOL IN LABORATORY TEST SYSTEMS

EPA Science Inventory

Shake flask and microcosm studies were conducted to determine the fate of parachlorophenol (p-CP) in water and sediment systems and the role of sediment and nonsediment surfaces in the biodegradation process. Biodegradation of p-CP in estuarine water samples in shake flasks was s...

A Volumetric Flask as a Projector

ERIC Educational Resources Information Center

Limsuwan, P.; Asanithi, P.; Thongpool, V.; Piriyawong, V.; Limsuwan, S.

2012-01-01

A lens based on liquid in the confined volume of a volumetric flask was presented as a potential projector to observe microscopic floating organisms or materials. In this experiment, a mosquito larva from a natural pond was selected as a demonstration sample. By shining a light beam from a laser pointer of any visible wavelength through the…

Process for casting hard-faced, lightweight camshafts and other cylindrical products

DOEpatents

Hansen, Jeffrey S.; Turner, Paul C.; Argetsinger, Edward R.; Wilson, Rick D.

1996-01-01

A process for casting a hard-faced cylindrical product such as an automobile camshaft includes the steps of: (a) preparing a composition formed from a molten base metal and an additive in particle form and having a hardness value greater than the hardness value of the base metal; (b) introducing the composition into a flask containing a meltable pattern of a cylindrical product such as an automobile camshaft to be manufactured and encased in sand to allow the composition to melt the pattern and assume the shape of the pattern within the sand; and (c) rotating the flask containing the pattern about the longitudinal axes of both the flask and the pattern as the molten base metal containing the additive in particle form is introduced into the flask to cause particles of the additive entrained in the molten base metal to migrate by centrifugal action to the radial extremities of the pattern and thereby provide a cylindrical product having a hardness value greater at it's radial extremities than at its center when the molten base metal solidifies.

Sensory Flask Cells in Sponge Larvae Regulate Metamorphosis via Calcium Signaling.

PubMed

Nakanishi, Nagayasu; Stoupin, Daniel; Degnan, Sandie M; Degnan, Bernard M

2015-12-01

The Porifera (sponges) is one of the earliest phyletic lineages to branch off the metazoan tree. Although the body-plan of sponges is among the simplest in the animal kingdom and sponges lack nervous systems that communicate environmental signals to other cells, their larvae have sensory systems that generate coordinated responses to environmental cues. In eumetazoans (Cnidaria and Bilateria), the nervous systems of larvae often regulate metamorphosis through Ca(2+)-dependent signal transduction. In sponges, neither the identity of the receptor system that detects an inductive environmental cue (hereafter "metamorphic cues") nor the signaling system that mediates settlement and metamorphosis are known. Using a combination of behavioral assays and surgical manipulations, we show here that specialized epithelial cells-referred to as flask cells-enriched in the anterior third of the Amphimedon queenslandica larva are most likely to be the sensory cells that detect the metamorphic cues. Surgical removal of the region enriched in flask cells in a larva inhibits the initiation of metamorphosis. The flask cell has an apical sensory apparatus with a cilium surrounded by an apical F-actin-rich protrusion, and numerous vesicles, hallmarks of eumetazoan sensory-neurosecretory cells. We demonstrate that these flask cells respond to metamorphic cues by elevating intracellular Ca(2+) levels, and that this elevation is necessary for the initiation of metamorphosis. Taken together, these analyses suggest that sponge larvae have sensory-secretory epithelial cells capable of converting exogenous cues into internal signals via Ca(2+)-mediated signaling, which is necessary for the initiation of metamorphosis. Similarities in the morphology, physiology, and function of the sensory flask cells in sponge larvae with the sensory/neurosecretory cells in eumetazoan larvae suggest this sensory system predates the divergence of Porifera and Eumetazoa. © The Author 2015. Published by Oxford University Press on behalf of the Society for Integrative and Comparative Biology. All rights reserved. For permissions please email: journals.permissions@oup.com.

The O-mannosylation and production of recombinant APA (45/47 KDa) protein from Mycobacterium tuberculosis in Streptomyces lividans is affected by culture conditions in shake flasks.

PubMed

Gamboa-Suasnavart, Ramsés A; Valdez-Cruz, Norma A; Cordova-Dávalos, Laura E; Martínez-Sotelo, José A; Servín-González, Luis; Espitia, Clara; Trujillo-Roldán, Mauricio A

2011-12-20

The Ala-Pro-rich O-glycoprotein known as the 45/47 kDa or APA antigen from Mycobacterium tuberculosis is an immunodominant adhesin restricted to mycobacterium genus and has been proposed as an alternative candidate to generate a new vaccine against tuberculosis or for diagnosis kits. In this work, the recombinant O-glycoprotein APA was produced by the non-pathogenic filamentous bacteria Streptomyces lividans, evaluating three different culture conditions. This strain is known for its ability to produce heterologous proteins in a shorter time compared to M. tuberculosis. Three different shake flask geometries were used to provide different shear and oxygenation conditions; and the impact of those conditions on the morphology of S. lividans and the production of rAPA was characterized and evaluated. Small unbranched free filaments and mycelial clumps were found in baffled and coiled shake flasks, but one order of magnitude larger pellets were found in conventional shake flasks. The production of rAPA is around 3 times higher in small mycelia than in larger pellets, most probably due to difficulties in mass transfer inside pellets. Moreover, there are four putative sites of O-mannosylation in native APA, one of which is located at the carboxy-terminal region. The carbohydrate composition of this site was determined for rAPA by mass spectrometry analysis, and was found to contain different glycoforms depending on culture conditions. Up to two mannoses residues were found in cultures carried out in conventional shake flasks, and up to five mannoses residues were determined in coiled and baffled shake flasks. The shear and/or oxygenation parameters determine the bacterial morphology, the productivity, and the O-mannosylation of rAPA in S. lividans. As demonstrated here, culture conditions have to be carefully controlled in order to obtain recombinant O-glycosylated proteins with similar "quality" in bacteria, particularly, if the protein activity depends on the glycosylation pattern. Furthermore, it will be an interesting exercise to determine the effect of shear and oxygen in shake flasks, to obtain evidences that may be useful in scaling-up these processes to bioreactors. Another approach will be using lab-scale bioreactors under well-controlled conditions, and study the impact of those on rAPA productivity and quality.

Long-term greenhouse gas measurements from aircraft

NASA Astrophysics Data System (ADS)

Karion, A.; Sweeney, C.; Wolter, S.; Newberger, T.; Chen, H.; Andrews, A.; Kofler, J.; Neff, D.; Tans, P.

2013-03-01

In March 2009 the NOAA/ESRL/GMD Carbon Cycle and Greenhouse Gases Group collaborated with the US Coast Guard (USCG) to establish the Alaska Coast Guard (ACG) sampling site, a unique addition to NOAA's atmospheric monitoring network. This collaboration takes advantage of USCG bi-weekly Arctic Domain Awareness (ADA) flights, conducted with Hercules C-130 aircraft from March to November each year. Flights typically last 8 h and cover a large area, traveling from Kodiak up to Barrow, Alaska, with altitude profiles near the coast and in the interior. NOAA instrumentation on each flight includes a flask sampling system, a continuous cavity ring-down spectroscopy (CRDS) carbon dioxide (CO2)/methane (CH4)/carbon monoxide (CO)/water vapor (H2O) analyzer, a continuous ozone analyzer, and an ambient temperature and humidity sensor. Air samples collected in flight are analyzed at NOAA/ESRL for the major greenhouse gases and a variety of halocarbons and hydrocarbons that influence climate, stratospheric ozone, and air quality. We describe the overall system for making accurate greenhouse gas measurements using a CRDS analyzer on an aircraft with minimal operator interaction and present an assessment of analyzer performance over a three-year period. Overall analytical uncertainty of CRDS measurements in 2011 is estimated to be 0.15 ppm, 1.4 ppb, and 5 ppb for CO2, CH4, and CO, respectively, considering short-term precision, calibration uncertainties, and water vapor correction uncertainty. The stability of the CRDS analyzer over a seven-month deployment period is better than 0.15 ppm, 2 ppb, and 4 ppb for CO2, CH4, and CO, respectively, based on differences of on-board reference tank measurements from a laboratory calibration performed prior to deployment. This stability is not affected by variation in pressure or temperature during flight. We conclude that the uncertainty reported for our measurements would not be significantly affected if the measurements were made without in-flight calibrations, provided ground calibrations and testing were performed regularly. Comparisons between in situ CRDS measurements and flask measurements are consistent with expected measurement uncertainties for CH4 and CO, but differences are larger than expected for CO2. Biases and standard deviations of comparisons with flask samples suggest that atmospheric variability, flask-to-flask variability, and possible flask sampling biases may be driving the observed flask versus in situ CO2 differences rather than the CRDS measurements.

THE PRODUCTION OF DIPHTHERIA TOXIN

PubMed Central

Park, W. H.; Williams, A. W.

1896-01-01

Toxin of sufficient strength to kill a 400-gramme guinea-pig in three days and a half in a dose of 0·cubic centimetre developed in suitable bouillon, contained in ordinary Erlenmeyer flasks, within a period of twenty-four hours. In such boullon the toxin reached its greatest strength in from four to seven days (0·005 cubic centimetre killing a 500-gramme guinea-pig in three days). This period of time covered that of the greatest growth of the bacilli, as shown both by the appearance of the culture and by the number of colonies developing an agar plates. The bodies of the diphtheria bacili did not at any time contain toxin in cosiderable amounts. The type of growth of the bacili and the rapidity and extent of the production of toxin depended more on the reaction of the bouillon than upon any other single factor. The best results were obtained in bouillon which, after being neutralized to litmus, had about seven cubic centimetres of normal soda solution added to each litre. An excessive amount of either acid or alkali prevented the development of toxin. Strong toxin was produced in bouillon containing peptone ranging from one to ten per cent. The strength of toxin averaged greater in the two and four-per-cent peptone solutions than in the one-percent. When the stage of acid reaction was brief and the degree of acidity probably slight, strong toxin developed while the culture bouillon was still acid; but when the stage of acid reaction was prolonged, little if any toxin was produced until just before the fluid became alkaline. Glucose is deleterious to the growth of the diphtheria bacillus and to the production of toxin when it is present in sufficient amounts to cause by its disintegration too great a degree of acidity in the fluid culture. When the acid resulting from decomposition of glucose is neutralized by the addition of alkali the diphtheria bacilus again grows abundantly. Glucose is not present, at least as a rule, in sufficient amounts in the meat as obtained from the New York butchers to prevent the rapid production of strong toxin if the bouillon is made sufficiently alkaline. In our experiments, when other conditions were similar, the strength of the toxin was in proportion to the virulence and vigour of growth of the bacillus employed. PMID:19866791

21 CFR 177.1970 - Vinyl chloride-lauryl vinyl ether copolymers.

Code of Federal Regulations, 2014 CFR

2014-04-01

... of the flask at 150 °F for 2 hours using a hot plate while also maintaining gentle mechanical agitation. Filter the contents of the flask rapidly through No. 42 Whatman filter paper with the aid of suction. Transfer the filtrate to flat glass dishes that are warmed on a hot plate and evaporate the...

Misconceptions in the Exploding Flask Demonstration Resolved through Students' Critical Thinking

ERIC Educational Resources Information Center

Spierenburg, Rick; Jacobse, Leon; de Bruin, Iris; van den Bos, Daan J.; Vis, Dominique M.; Juurlink, Ludo B. F.

2017-01-01

As it connects to a large set of important fundamental ideas in chemistry and analytical techniques discussed in high school chemistry curricula, we review the exploding flask demonstration. In this demonstration, methanol vapor is catalytically oxidized by a Pt wire catalyst in an open container. The exothermicity of reactions occurring at the…

Using PETRIMES to estimate mercury deposits in California

USGS Publications Warehouse

Lee, P.J.; Singer, D.A.

1994-01-01

In this article, we examine the use of an unconventional procedure, PETRIMES, to estimate mineral resources of mercury deposits in California. The study, which is based on the nonparametric discovery process model and Q-Q plots, suggests that a lognormal distribution is appropriate for the mercury deposits in California. The results of the assessment are summarized as follows: (1) the total number of mercury deposits in the population is approximately 165; (2) the median value of the largest undiscovered deposit size is 487 flasks; (3) the mean of the remaining mercury potential is 2,500 flasks; and (4) the population resource ranges from 1,040,000 to 4,300,000 flasks (at a 0.9 probability level). ?? 1994 Oxford University Press.

Volume to dissolve applied dose (VDAD) and apparent dissolution rate (ADR): tools to predict in vivo bioavailability from orally applied drug suspensions.

PubMed

Muenster, Uwe; Pelzetter, Christian; Backensfeld, Thomas; Ohm, Andreas; Kuhlmann, Thomas; Mueller, Hartwig; Lustig, Klemens; Keldenich, Jörg; Greschat, Susanne; Göller, Andreas H; Gnoth, Mark Jean

2011-08-01

Low solubility of drug candidates generated in research contributes to their elimination during subsequent development due to insufficient oral bioavailability (BA) of crystalline compound. Therefore, the purpose of the study was to identify critical in vitro solubility and dissolution parameter that would predict critical in vivo dissolution by means of in vitro-in vivo correlation. Thermodynamic solubility and apparent dissolution rate (ADR) were determined using the shake-flask method and mini-flow-through-cell, respectively. Oral BA studies in rats and humans were conducted from drug solution and suspension/tablets. Relative BA was calculated using F(rel) [%]=AUC(suspension)/AUC(solution)*100, representing a measure of in vivo dissolution. Roughly, F(rel) rat >50% translates into F(rel) human of >90%. Both, ADR and log volume to dissolve applied dose (VDAD), when plotted against F(rel) rat, revealed certain threshold levels, (ADR, ∼150-200 μg of compound dissolved under respective assay conditions; VDAD, ∼100-500 ml/kg) which translate into F(rel) in rats of >50%. Thus, assuming that F(rel)>50% in rats is indicative of sufficient in vivo dissolution in humans after oral application, drugs should exhibit a VDAD of ∼100-500 ml/kg or less in aqueous media to avoid insufficient or varying drug absorption. Copyright © 2011 Elsevier B.V. All rights reserved.

Tannase sequence from a xerophilic Aspergillus niger Strain and production of the enzyme in Pichia pastoris.

PubMed

Fuentes-Garibay, José Antonio; Aguilar, Cristóbal Noé; Rodríguez-Herrera, Raúl; Guerrero-Olazarán, Martha; Viader-Salvadó, José María

2015-05-01

Tannin acyl hydrolases, or tannases (EC 3.1.1.20), are enzymes with potential biotechnological applications. In this work, we describe the gene and amino acid sequences of the tannase from Aspergillus niger GH1. In addition, we engineered Pichia pastoris strains to produce and secrete the enzyme, and the produced tannase was characterized biochemically. The nucleotide sequence of mature tannase had a length of 1,686 bp, and encodes a protein of 562 amino acids. A molecular model of mature A. niger GH1 tannase showed the presence of two structural domains, one with an α/β-hydrolase fold and one lid domain that covers the catalytic site, likely being residues Ser-196, Asp-448, and His-494 the putative catalytic triad, which are connected by a disulfide bond between the neighboring cysteines, Cys-195 and Cys-495. A 120-ml shake flask culture with a constructed recombinant P. pastoris strain showed extracellular tannase activity at 48 h induction of 0.57 U/ml. The produced tannase was N-glycosylated, consisted of two subunits, likely linked by a disulfide bond, and had an optimum pH of 5.0 and optimum temperature of 20 °C. These biochemical properties differed from those of native A. niger GH1 tannase. The recombinant tannase could be suitable for food and beverage applications.

Expression, purification and characterization of a phyAm-phyCs fusion phytase*

PubMed Central

Zou, Li-kou; Wang, Hong-ning; Pan, Xin; Tian, Guo-bao; Xie, Zi-wen; Wu, Qi; Chen, Hui; Xie, Tao; Yang, Zhi-rong

2008-01-01

The phyAm gene encoding acid phytase and optimized neutral phytase phyCs gene were inserted into expression vector pPIC9K in correct orientation and transformed into Pichia pastoris in order to expand the pH profile of phytase and decrease the cost of production. The fusion phytase phyAm-phyCs gene was successfully overexpressed in P. pastoris as an active and extracellular phytase. The yield of total extracellular fusion phytase activity is (25.4±0.53) U/ml at the flask scale and (159.1±2.92) U/ml for high cell-density fermentation, respectively. Purified fusion phytase exhibits an optimal temperature at 55 °C and an optimal pH at 5.5~6.0 and its relative activity remains at a relatively high level of above 70% in the range of pH 2.0 to 7.0. About 51% to 63% of its original activity remains after incubation at 75 °C to 95 °C for 10 min. Due to heavy glycosylation, the expressed fusion phytase shows a broad and diffuse band in SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). After deglycosylation by endoglycosidase H (EndoHf), the enzyme has an apparent molecular size of 95 kDa. The characterization of the fusion phytase was compared with those of phyCs and phyAm. PMID:18600783

Adaptation and Molecular Characterization of Two Malaysian Very Virulent Infectious Bursal Disease Virus Isolates Adapted in BGM-70 Cell Line

PubMed Central

Lawal, Nafi'u; Arshad, Siti Suri

2017-01-01

Two Malaysian very virulent infectious bursal disease virus (vvIBDV) strains UPM0081 and UPM190 (also known as UPMB00/81 and UPM04/190, respectively) isolated from local IBD outbreaks were serially passaged 12 times (EP12) in specific pathogen free (SPF) chicken embryonated eggs (CEE) by chorioallantoic membrane (CAM) route. The EP12 isolate was further adapted and serially propagated in BGM-70 cell line up to 20 passages (P20). Characteristic cytopathic effects (CPEs) were subtly observed at P1 in both isolates 72 hours postinoculation (pi). The CPE became prominent at P5 with cell rounding, cytoplasmic vacuoles, granulation, and detachment from flask starting from day 3 pi, up to 7 days pi with titers of 109.50 TCID50/mL and log109.80 TCID50/mL for UPM0081 and UPM190, respectively. The CPE became subtle at P17 and disappeared by P18 and P19 for UPM0081 and UPM190, respectively. However, the presence of IBDV was confirmed by immunoperoxidase, immunofluorescence, and RT-PCR techniques. Phylogenetic analysis showed that these two isolates were of the vvIBDV. It appears that a single mutation of UPM190 and UPM0081 IBDV isolates at D279N could facilitate vvIBDV strain adaptability in CEE and BGM-70 cultures. PMID:29230245

Laboratory production of human prolactin from CHO cells adapted to serum-free suspension culture.

PubMed

Arthuso, Fernanda Santos; Bartolini, Paolo; Soares, Carlos Roberto Jorge

2012-08-01

Human prolactin (hPRL) is a polypeptide with 199 amino acids and a molecular mass of 23 kDa. Previously, a eukaryotic hPRL expression vector was used to transfect Chinese hamster ovary (CHO) cells: this work describes a fast and practical laboratory adaptation of these transfected cells, in ~40 days, to grow in suspension in serum-free medium. High cell densities of up to 4.0 × 10(6) cell/ml were obtained from spinner flask cultures and a stable and continuous production process was developed for at least 30 days. Two harvesting strategies were set up, 50 or 100 % of the total conditioned medium being collected daily and replaced by fresh culture medium. The volumetric productivity was 5-7 μg hPRL/ml, as determined directly in the collected medium via reversed-phase HPLC (RP-HPLC). A two-step process based on a cationic exchanger followed by size exclusion chromatography was applied to obtain purified hPRL from conditioned medium. Two hPRL isoforms, non-glycosylated and glycosylated, could also be separated by high-performance size-exclusion chromatography (HPSEC) and, when analyzed by RP-HPLC, HPSEC, Western blotting, and bioassay, were found to be comparable to the World Health Organization International Reference Reagent of hPRL. These results are useful for the practical scale-up to the pilot and industrial scale of a bioprocess based on CHO cell culture.

Simplified production and concentration of HIV-1-based lentiviral vectors using HYPERFlask vessels and anion exchange membrane chromatography

PubMed Central

Kutner, Robert H; Puthli, Sharon; Marino, Michael P; Reiser, Jakob

2009-01-01

Background During the past twelve years, lentiviral (LV) vectors have emerged as valuable tools for transgene delivery because of their ability to transduce nondividing cells and their capacity to sustain long-term transgene expression in target cells in vitro and in vivo. However, despite significant progress, the production and concentration of high-titer, high-quality LV vector stocks is still cumbersome and costly. Methods Here we present a simplified protocol for LV vector production on a laboratory scale using HYPERFlask vessels. HYPERFlask vessels are high-yield, high-performance flasks that utilize a multilayered gas permeable growth surface for efficient gas exchange, allowing convenient production of high-titer LV vectors. For subsequent concentration of LV vector stocks produced in this way, we describe a facile protocol involving Mustang Q anion exchange membrane chromatography. Results Our results show that unconcentrated LV vector stocks with titers in excess of 108 transduction units (TU) per ml were obtained using HYPERFlasks and that these titers were higher than those produced in parallel using regular 150-cm2 tissue culture dishes. We also show that up to 500 ml of an unconcentrated LV vector stock prepared using a HYPERFlask vessel could be concentrated using a single Mustang Q Acrodisc with a membrane volume of 0.18 ml. Up to 5.3 × 1010 TU were recovered from a single HYPERFlask vessel. Conclusion The protocol described here is easy to implement and should facilitate high-titer LV vector production for preclinical studies in animal models without the need for multiple tissue culture dishes and ultracentrifugation-based concentration protocols. PMID:19220915

A simple and efficient method for generating Nurr1-positive neuronal stem cells from human wisdom teeth (tNSC) and the potential of tNSC for stroke therapy.

PubMed

Yang, Kuo-Liang; Chen, Mei-Fang; Liao, Chia-Hsin; Pang, Cheng-Yoong; Lin, Py-Yu

2009-01-01

We have isolated human neuronal stem cells from exfoliated third molars (wisdom teeth) using a simple and efficient method. The cultured neuronal stem cells (designated tNSC) expressed embryonic and adult stem cell markers, markers for chemotatic factor and its corresponding ligand, as well as neuron proteins. The tNSC expressed genes of Nurr1, NF-M and nestin. They were used to treat middle cerebral artery occlusion (MCAO) surgery-inflicted Sprague-Dawley (SD) rats to assess their therapeutic potential for stroke therapy. For each tNSC cell line, a normal human impacted wisdom tooth was collected from a donor with consent. The tooth was cleaned thoroughly with normal saline. The molar was vigorously shaken or vortexed for 30 min in a 50-mL conical tube with 15-20mL normal saline. The mixture of dental pulp was collected by centrifugation and cultured in a 25-cm(2) tissue culture flask with 4-5mL Medium 199 supplemented with 5-10% fetal calf serum. The tNSC harvested from tissue culture, at a concentration of 1-2x10(5), were suspended in 3 microL saline solution and injected into the right dorsolateral striatum of experimental animals inflicted with MCAO. Behavioral measurements of the tNSC-treated SD rats showed a significant recovery from neurologic dysfunction after MCAO treatment. In contrast, a sham group of SD rats failed to recover from the surgery. Immunohistochemistry analysis of brain sections of the tNSC-treated SD rats showed survival of the transplanted cells. These results suggest that adult neuronal stem cells may be procured from third molars, and tNSC thus cultivated have potential for treatment of stroke-inflicted rats.

Isolation of marine fungi Aspergillus sp. and its in vitro antifouling activity against marine bacteria.

PubMed

Thiyagarajan, Santhananmari; Bavya, Manoharan; Jamal, Alruwaili

2016-09-01

Biofouling is considered as a main issue of concern in aquatic environment causing severe economic loss and pollution. The aim of the present study was to isolate marine fungus antagonistic to biofouling bacteria and to define antifouling compounds present in it. Using standard plate method five predominant biofouling bacteria viz., Methylococcus sp., Flavobacterium sp., Marinococcus sp., Serratia sp. and Pseudomonas sp. were isolated from marine solid substances on Zobell's agar. Tolerance range of these bacteria to NaCl was 2-10%. Isolation of fungi from mangrove and estuarine sediments and their screening identified Aspergillus sp. EF4 as a potential isolate. This isolate caused inhibition of all the five test bacterial cultures measuring zone diameters respectively of 11, 16, 12, 13 and 11mm.? Subsequent to submerged fermentation using shaking flask method this fungus produced bioactive compounds within 5 days. The culture parameters optimized were raffinose as carbon source, yeast extract as lone nitrogen source, pH up to 9.0 and temperature up to 40?C. Antifouling compounds of culture filtrate were separated and detected by a three-step procedure involving thin layer chromatography, bioautography and preparative TLC. The in vitro assay involving glass slide-wooden stick-biofilm method revealed that these compounds could cause inhibition and destruction of bacteria to an extent of 2.16 x 104 CFU ml-1 and 2.46 x 104 CFU ml-1 respectively while growth of bacteria in control beaker was enumerated to be 4.41 x 104 CFU ml-1. High performance liquid chromatography of culture filtrate indicated probable principal antifouling compound as Fumonisin B2. Isolation of antagonistic marine fungus from Indian coast and detection of its antifouling compound would help in planning effective strategies for controlling biofouling in marine environment.

Production and characterization of active recombinant interleukin-12/eGFP fusion protein in stably-transfected DF1 chicken cells.

PubMed

Wu, Hsing Chieh; Chen, Yu San; Shen, Pin Chun; Shien, Jui Hung; Lee, Long Huw; Chiu, Hua Hsien

2015-01-01

The adjuvant activity of chicken interleukin-12 (chIL-12) protein has been described as similar to that of mammalian IL-12. Recombinant chIL-12 can be produced using several methods, but chIL-12 production in eukaryotic cells is lower than that in prokaryotic cells. Stimulating compounds, such as dimethyl sulfoxide (DMSO), can be added to animal cell cultures to overcome this drawback. In this study, we constructed a cell line, DF1/chIL-12 which stably expressed a fusion protein, chIL-12 and enhanced green fluorescent protein (eGFP) connected by a (G4 S)3 linker sequence. Fusion protein production was increased when cells were cultured in the presence of DMSO. When 1 × 10(6) DF1/chIL-12 cells were inoculated in a T-175 flask containing 30 mL of media, incubated for 15 h, and further cultivated in the presence of 4% DMSO for 48 h, the production of total fusion protein was mostly enhanced compared with the production of total fusion protein by using cell lysates induced with DMSO at other concentrations. The concentrations of the unpurified and purified total fusion proteins in cell lysates were 2,781 ± 2.72 ng mL(-1) and 2,207 ± 3.28 ng mL(-1) , respectively. The recovery rate was 79%. The fusion protein stimulated chicken splenocytes to produce IFN-γ, which was measured using an enzyme-linked immunosorbent assay, in the culture supernatant, indicating that treating DF1/chIL-12 cells with DMSO or producing chIL-12 in a fusion protein form does not have adverse effects on the bioactivity of chIL-12. © 2015 American Institute of Chemical Engineers.

Revisit: A Surprising Demonstration of Total Internal Reflection

ERIC Educational Resources Information Center

Lee, Jiwon; Cha, Yu Wha; Jung, Yeon Su; Oh, Eun Ju; Moon, Ye Lin; Kim, Jung Bog

2016-01-01

Melton demonstrated a surprising disappearance using total internal reflection. When he put a Florence flask filled with marbles into a water tank and looked straight down from directly above the flask, he was only able to see marbles above a certain water level. When he added more water into the tank above the top line of the marbles, all of the…

Wider-Opening Dewar Flasks for Cryogenic Storage

NASA Technical Reports Server (NTRS)

Ruemmele, Warren P.; Manry, John; Stafford, Kristin; Bue, Grant; Krejci, John; Evernden, Bent

2010-01-01

Dewar flasks have been proposed as containers for relatively long-term (25 days) storage of perishable scientific samples or other perishable objects at a temperature of 175 C. The refrigeration would be maintained through slow boiling of liquid nitrogen (LN2). For the purposes of the application for which these containers were proposed, (1) the neck openings of commercial off-the-shelf (COTS) Dewar flasks are too small for most NASA samples; (2) the round shapes of the COTS containers give rise to unacceptably low efficiency of packing in rectangular cargo compartments; and (3) the COTS containers include metal structures that are too thermally conductive, such that they cannot, without exceeding size and weight limits, hold enough LN2 for the required long-term-storage. In comparison with COTS Dewar flasks, the proposed containers would be rectangular, yet would satisfy the long-term storage requirement without exceeding size and weight limits; would have larger neck openings; and would have greater sample volumes, leading to a packing efficiency of about double the sample volume as a fraction of total volume. The proposed containers would be made partly of aerospace- type composite materials and would include vacuum walls, multilayer insulation, and aerogel insulation.

The radial transmission line as a broad-band shielded exposure system for microwave irradiation of large numbers of culture flasks.

PubMed

Moros, E G; Straube, W L; Pickard, W F

1999-01-01

The problem of simultaneously exposing large numbers of culture flasks at nominally equivalent incident power densities and with good thermal control is considered, and the radial transmission line (RTL) is proposed as a solution. The electromagnetic design of this structure is discussed, and an extensively bench-tested realization is described. Referred to 1 W of net forward power, the following specific absorption rate (SAR) data were obtained: at 835.62 MHz, 16.0+/-2.5 mW/kg (mean+/-SD) with range (11-22); at 2450 MHz, 245+/-50 mW/kg with range (130-323). Radio-frequency interference from an RTL driven at roughly 100 W is so low as to be compatible with a cellular base station only 500 m distant. To avoid potential confounding by temperature differences among as many as 144 T-75 flasks distributed over 9 RTLs (six irradiates and three shams), temperature within all flasks was controlled to 37.0+/-0.3 degrees C. Experience with over two years of trouble-free operation suggests that the RTL offers a robust, logistically friendly, and environmentally satisfactory solution to the problem of large-scale in vitro experiments in bioelectromagnetics.

A vacuum-operated pore-water extractor for estuarine and freshwater sediments

USGS Publications Warehouse

Winger, Parley V.; Lasier, Peter J.

1991-01-01

A vacuum-operated pore-water extractor for estuarine and freshwater sediments was developed and constructed from a fused-glass air stone attached with aquarium airline tubing to a 30 or 60 cc polypropylene syringe. Pore water is extracted by inserting the air stone into the sediment and creating a vacuum by retracting and bracing the syringe plunger. A hand-operated vacuum pump attached to a filtration flask was also evaluated as an alternative vacuum source. The volume and time to extract pore water varies with the number of devices and the sediment particle size. Extraction time is longer for fine sediments than for sandy sediments. Four liters of sediment generally yield between 500 and 1,500 mL of pore water. The sediment that surrounds and accumulates on the air stone acts as a filter, and, except for the first few milliliters, the collected pore water is clear. Because there is no exposure to air or avenue for escape, volatile compounds andin situ characteristics are retained in the extracted pore water.

An Innovative Very Low Thermal Power Waste Heat Recovery System for Thermal Control of Deep Space Missions: A Thermal Flask in Space

NASA Technical Reports Server (NTRS)

Bhandari, Pradeep

2015-01-01

Future missions to deep space, such as those to the outer planets (Jupiter, Saturn, etc.), which would rely on solar photovoltaic power, would need extremely large solar arrays to produce sufficient power for their operations because solar intensity is so low at those locations. Hence any additional power that would be needed for thermal control is extremely limited. Previous deep space missions like Juno (to Jupiter) required almost 200 W of electrical power for thermal control. This is prohibitively large for many future mission concepts, and leads to them needing very large solar arrays. For Saturn, where the solar flux is 1/4th the flux at Jupiter, this would entail an extremely large increase in the solar array size to accommodate the need for thermal survival power, which would be prohibitively large in size and mass, and very expensive. Hence there is a need to come up with a thermal architecture and design options that would not need such prohibitively large thermal power levels. One solution relies on harvesting the pre-existing waste heat from all the heat dissipation that would be present from operation of electronics, instruments, etc. for their own functionality. For example, for a generic Saturn mission, the various electronics would already dissipate about 200 Watts of heat that is simply "thrown away" to space from the spacecraft surfaces. The amount of thermal power that would be required for the safe thermal control of components within the spacecraft in deep space would be roughly of this magnitude for this class of spacecraft. So it makes good sense to try to harvest the waste heat and employ it to maintain the temperatures of all the components within their allowable limits. In particular, propulsion systems typically need to be kept above their freezing limits, around room temperature (15 C). Electronics needs to be kept typically above -40 C and batteries above -20 C. The next question becomes how to harvest this waste heat and direct it to the components that would need it for their survival. The proposed system utilizes a mechanically pumped, single phase fluid loop to pick up the waste heat from components attached to this loop's tubing and then directed to a thermal flask that has tubing attached to it. The thermal flask is cylindrically shaped and contains essentially all systems and components in the spacecraft within it, with the exception of the solar array, antennae, thrusters and various apertures of instruments, etc. to allow them an unobstructed view of space. Waste heat from the heat-dissipating components warms up the fluid and is carried to the flask surface and deposited on it via the fluid loop's flow. The entire flask is covered with Multi-Layered Insulation (MLI) to minimize the heat loss from the flask and allow it to remain warm. Hence the flask essentially creates a thermal environment within which the spacecraft components reside. The temperature of the components within the flask is then essentially the same as the temperature of the flask. This approach could be a very enabling feature for deep space missions. This paper describes the approach utilized for this thermal architecture, along with its mechanical and implementation aspects. Additionally it will compare and contrast this approach with the more conventional solutions utilized earlier.

40 CFR 63.2164 - If I monitor brew ethanol, what are my monitoring installation, operation, and maintenance...

Code of Federal Regulations, 2010 CFR

2010-07-01

... batch averages computed from two or more data points over each 1-hour period, except during periods when... performed. During these periods, a valid hour of data shall consist of at least one data point representing... volumetric flask and place the flask on a balance. Tare the balance. Weigh 2.3670 grams of the dry (anhydrous...

Development of Pattern Recognition Techniques for the Evaluation of Toxicant Impacts to Multispecies Systems

DTIC Science & Technology

1993-06-18

the exception. In the Standardized Aquatic Microcosm and the Mixed Flask Culture (MFC) microcosms, multivariate analysis and clustering methods...rule rather than the exception. In the Standardized Aquatic Microcosm and the Mixed Flask Culture (MFC) microcosms, multivariate analysis and...experiments using two microcosm protocols. We use nonmetric clustering, a multivariate pattern recognition technique developed by Matthews and Heame (1991

Optimization of CO2 Surface Flux using GOSAT Total Column CO2: First Results for 2009-2010

NASA Astrophysics Data System (ADS)

Basu, S.; Houweling, S.

2011-12-01

Constraining surface flux estimates of CO2 using satellite measurements has been one of the long-standing goals of the atmospheric inverse modeling community. We present the first results of inverting GOSAT total column CO2 measurements for obtaining global monthly CO2 flux maps over one year (June 2009 to May 2010). We use the SRON RemoTeC retrieval of CO2 for our inversions. The SRON retrieval has been shown to have no bias when compared to TCCON total column measurements, and latitudinal gradients of the retrieved CO2 are consistent with gradients deduced from the surface flask network [Butz et al, 2011]. This makes this retrieval an ideal candidate for atmospheric inversions, which are highly sensitive to spurious gradients. Our inversion system is analogous to the CarbonTracker (CT) data assimilation system; it is initialized with the prior CO2 fluxes of CT, and uses the same atmospheric transport model, i.e., TM5. The two major differences are (a) we add GOSAT CO2 data to the inversion in addition to flask data, and (b) we use a 4DVAR optimization system instead of a Kalman filter. We compare inversions using (a) only GOSAT total column CO2 measurements, (b) only surface flask CO2 measurements, and (c) the joint data set of GOSAT and surface flask measurements. We validate GOSAT-only inversions against the NOAA surface flask network and joint inversions against CONTRAIL and other aircraft campaigns. We see that inverted fluxes from a GOSAT-only inversion are consistent with fluxes from a stations-only inversion, reaffirming the low biases in SRON retrievals. From the joint inversion, we estimate the amount of added constraints upon adding GOSAT total column measurements to existing surface layer measurements.

Trophic interactions in the plankton of Malyi Sevan in July, 1984

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

Gulelmacher, B.L.; Simonyan, A.A.

1986-09-01

This paper describes some studies of the trophic interactions in the plankton in Lchashenskaya Bay in Malyi Sevan in July, 1984. The goal was to study the feeding characteristics of the major crustacean species and their influence on the phytoplankton. To study the effect of the planktonic crustaceans on the phytoplankton, a flask with an added 50 Acanthodiaptomus and a flask without crustaceans were exposed for 24 h after which a radioactive carbon isotope was added to them in the form of sodium carbonate. The radioactivity of the filters and working solutions was measured and autoradiographic preparations were made. Itmore » is shown that the more than threefold drop in the quantity of algae in a flask resulting from crustacean feeding did not lead to a significant decrease in primary production.« less

Atmospheric CO2 Records from Sites in the Scripps Institution of Oceanography (SIO) Air Sampling Network (1985 - 2007)

DOE Data Explorer

Keeling, R. F. [Scripps Institution of Oceanography (SIO) University of California, La Jolla, California (USA); Piper, S. C. [Scripps Institution of Oceanography (SIO) University of California, La Jolla, California (USA); Bollenbacher, A. F. [Scripps Institution of Oceanography (SIO) University of California, La Jolla, California (USA); Walker , J. S. [Scripps Institution of Oceanography (SIO) University of California, La Jolla, California (USA)

2008-05-01

At Alert weekly air samples are collected in 5-L evacuated glass flasks exposed in triplicate. Flasks are returned to the SIO for CO2 determinations, which are made using an Applied Physics Corporation nondispersive infrared gas analyzer. In May 1983, the CO2-in-N2 calibration gases were replaced with CO2-in-air calibration gases, which are currently used (Keeling et al. 2002). Data are in terms of the Scripps "03A" calibration scale. On the basis of flask samples collected at Alert and analyzed by SIO, the annual average of the fitted monthly concentrations CO2 rose from 348.48 ppmv in 1986 to 384.84 ppmv in 2007. This represents an average annual growth rate of 1.73 ppmv per year at Alert.

Cultivation of recombinant Chinese hamster ovary cells grown as suspended aggregates in stirred vessels.

PubMed

Han, Yi; Liu, Xing-Mao; Liu, Hong; Li, Shi-Chong; Wu, Ben-Chuan; Ye, Ling-Ling; Wang, Qu-Wei; Chen, Zhao-Lie

2006-11-01

Recombinant Chinese hamster ovary (rCHO) cells capable of producing a prourokinase mutant (mPro-uk) grown as suspended aggregates in stirred vessels were described and characterized. The addition of chitosan to a mixture of DMEM and Ham's F12 (D-MEM/F-12) medium promoted cell aggregation and spheroid formation efficiently. Multicellular aggregates formed immediately after the rCHO cells were inoculated into the chitosan-added medium, and the mean diameter of the cell aggregates reflecting the aggregate size increased with culture time, shifting from 65 to 163 mum after 2 and 9 d of culture in spinner flasks. No significant difference in the metabolism performance of the rCHO cells was observed between suspended aggregates and anchored monolayers. However, the cells cultured as suspended aggregates showed a marked decrease in growth rate as evaluated from specific growth rate (mu). Replacing D-MEM/F-12 medium with CD 293 medium caused compact spherical cell aggregates to dissociate into small irregular aggregates and single cells without apparent effects on cell performance in subcultures. The perfusion culture of the rCHO cells grown as suspended aggregates in a 2-l stirred tank bioreactor for 15 d resulted in a maximum viable cell density of 5.6 x 10(6) cells ml(-1) and an mPro-uk concentration of about 2.6 x 10(3) IU ml(-1), and cell viability was remained at roughly 90% during the entire run.

The Flask Security Architecture: System Support for Diverse Security Policies

DTIC Science & Technology

2006-01-01

Flask microkernel -based operating sys­ tem, that successfully overcomes these obstacles to pol- icy flexibility. The cleaner separation of mechanism and...other object managers in the system to en- force those access control decisions. Although the pro­ totype system is microkernel -based, the security...mecha­ nisms do not depend on a microkernel architecture and will easily generalize beyond it. The resulting system provides policy flexibility. It sup

Biofilm lifestyle enhances diesel bioremediation and biosurfactant production in the Antarctic polyhydroxyalkanoate producer Pseudomonas extremaustralis.

PubMed

Tribelli, Paula M; Di Martino, Carla; López, Nancy I; Raiger Iustman, Laura J

2012-09-01

Diesel is a widely distributed pollutant. Bioremediation of this kind of compounds requires the use of microorganisms able to survive and adapt to contaminated environments. Pseudomonas extremaustralis is an Antarctic bacterium with a remarkable survival capability associated to polyhydroxyalkanoates (PHAs) production. This strain was used to investigate the effect of cell growth conditions--in biofilm versus shaken flask cultures--as well as the inocula characteristics associated with PHAs accumulation, on diesel degradation. Biofilms showed increased cell growth, biosurfactant production and diesel degradation compared with that obtained in shaken flask cultures. PHA accumulation decreased biofilm cell attachment and enhanced biosurfactant production. Degradation of long-chain and branched alkanes was observed in biofilms, while in shaken flasks only medium-chain length alkanes were degraded. This work shows that the PHA accumulating bacterium P. extremaustralis can be a good candidate to be used as hydrocarbon bioremediation agent, especially in extreme environments.

Atmospheric CO2 Concentrations from Aircraft for 1972-1981, CSIRO Monitoring Program

DOE Data Explorer

Beardsmore, David J. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Victoria, Australia; Pearman, Graeme I. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Victoria, Australia

2012-01-01

From 1972 through 1981, air samples were collected in glass flasks from aircraft at a variety of latitudes and altitudes over Australia, New Zealand, and Antarctica. The samples were analyzed for CO2 concentrations with nondispersive infrared gas analysis. The resulting data contain the sampling dates, type of aircraft, flight number, flask identification number, sampling time, geographic sector, distance in kilometers from the listed distance measuring equipment (DME) station, station number of the radio navigation distance measuring equipment, altitude of the aircraft above mean sea level, sample analysis date, flask pressure, tertiary standards used for the analysis, analyzer used, and CO2 concentration. These data represent the first published record of CO2 concentrations in the Southern Hemisphere expressed in the WMO 1981 CO2 Calibration Scale and provide a precise record of atmospheric CO2 concentrations in the troposphere and lower stratosphere over Australia and New Zealand.

Atmospheric Carbon Dioxide Record from Flask Measurements at Lampedusa Island (May 1992- December 2000)

DOE Data Explorer

Chamard, Paolo [ENEA, GEM-CLIM, Rome, Italy; Ciattaglia, Luigi [ENEA, GEM-CLIM, Rome, Italy; di Sarra, Alcide [ENEA, GEM-CLIM, Rome, Italy; Monteleone, Francesco [ENEA, GEM-CLIM, Rome, Italy

2001-01-01

Lampedusa Station rests on a rocky seashore on the eastern tip of Lampedusa Island, located south of Sicily in the central Mediterranean Sea. Air samples at Lampedusa Station are collected each Friday in two pairs of 2-L glass flasks. Flasks are evacuated and then pressurized to 280-300 kPa with ambient air at the sampling site. Determinations of CO2 are made by using a Siemens Ultramat 5E nondispersive infrared gas analyzer. Water vapor is removed during the sampling process by means of a chemical trap (Magnesium Perchlorate). During the measurement, residual water vapor is removed by means of a cold trap at -70°C. From the inception of the monitoring program at Lampedusa Station, CO2-in-synthetic air calibration gases have been used. Values are relative to the dry air WMO mole fraction scale.

Use of saline waste water from demineralization of cheese whey for cultivation of Schizochytrium limacinum PA-968 and Japonochytrium marinum AN-4.

PubMed

Humhal, Tomas; Kastanek, Petr; Jezkova, Zuzana; Cadkova, Anna; Kohoutkova, Jana; Branyik, Tomas

2017-03-01

Saline waste water from demineralization of cheese whey was used as the main component of waste saline medium (WSM) for cultivation of thraustochytrids. The suitability of WSM for cultivation of Schizochytrium limacinum PA-968 and Japonochytrium marinum AN-4 was evaluated by comparison with cultivation on nutrient medium (NM) in shake flask and fermenter cultures. Biomass productivities achieved in WSM for the thraustochytrids were comparable with those in NM for both shake flask and fermenter cultures. The maximum total lipid content (56.71% dry cell weight) and docosahexaenoic acid productivity (0.86 g/L/day) were achieved by J. marinum AN-4 grown on WSM in shake flask and fermenter cultures, respectively. A cost estimate of WSM suggests that this medium could result in lower production costs for thraustochytrid biomass and lipids and contribute to the effective reduction in saline diary process waste water.

Influence of Nutrient Impregnated into Zeolite Addition on Anaerobic Digestion of Palm Oil Mill Effluent (POME)

NASA Astrophysics Data System (ADS)

Mellyanawaty, M.; Chusna, F. M. A.; Sudibyo, H.; Nurjanah, N.; Budhijanto, W.

2018-03-01

Palm oil mill effluent (POME) was wastewater generated from palm oil milling activities which was brownish liquid, acidic with pH 3-4, and contained soluble materials which were hazardous to the environment. It was characterized by high organic loading (COD 40,000–60,000 mg/L). According to its characteristics, POME was identified as a potential source to generate renewable energy through anaerobic digestion. In other words, a combination of wastewater treatment and renewable energy production would be an additional advantage to the palm oil industries. Methanogenesis was the rate limiting step in anaerobic digestion. In the conventional anaerobic digester, it required large reactors and long retention time. The addition of microbial immobilization media was to improve anaerobic reactor performance in term of higher organic removal and methane production. Additionally, better performance could lead to reduction of reactor volume and shorter retention time in high rate anaerobic digester. The loading of essential microorganism nutrient into the media might increase the affinity of bacteria to attach and grow on the media surface. Activating or inhibition effects of natural and modified zeolite addition in anaerobic digestion of POME was studied in batch reactors using erlenmeyer of 1,000 mL at COD concentrations of about 8,000 mg/L. Zeolite was impregnated with nickel and magnesium at concentrations of 0.0561 mg Ni/g zeolite and 0.0108 mg Mg/g zeolite. The effect of the different zeolite addition was determined by the measurement of soluble COD (sCOD), Volatile Fatty Acids (VFAs) and biogas production. Greater effect of modified zeolite was observed in zeolite impregnated with nickel with a 54% increase of biogas production. Meanwhile, the modified zeolite impregnated with magnesium had no positive impact to the methanogenic bacteria activities.

Studies of Cell-Mediated Immunity Against Immune Disorders Using Synthetic Peptides and Rotating Bioreactor System

NASA Technical Reports Server (NTRS)

Sastry, Jagannadha K.

1997-01-01

Our proposed experiments included: (1) immunzing mice with synthetic peptides; (2) preparing spleen and lymph node cells; (3) growing them under conventional conditions as well as in the rotatory vessel in appropriate medium reconstituting with synthetic peptides and/or cytokines as needed; and (4) comparing at regular time intervals the specific CTL activity as well as helper T-cell activity (in terms of both proliferative responses and cytokine production) using established procedures in my laboratory. We further proposed that once we demonstrated the merit of rotatory vessel technology to achieve desired results, these studies would be expanded to include immune cells from non-human primates (rhesus monkeys and chimpanzees) and also humans. We conducted a number of experiments to determine CTL induction by the synthetic peptides corresponding to antigenic proteins in HIV and HPV in different mouse strains that express MHC haplotypes H-2b or H-2d. We immunized mice with 100 ug of the synthetic peptide, suspended in sterile water, and emulsified in CFA (1:1). The immune lymph node cells obtained after 7 days were restimulated by culturing in T25 flask, HARV-10, or STLV-50, in the presence of the peptide at 20 ug/ml. The results from the 5'Cr-release assay consistently revealed complete abrogation of CTL activity of cells grown in the bioreactors (both HARV and STLV), while significant antigen-specific CTL activity was observed with cells cultured in tissue culture flasks. Thus, overall the data we generated in this study proved the usefulness of the NASA-developed developed technology for understanding the known immune deficiency during space travel. Additionally, this ex vivo microgravity technology since it mimics effectively the in vivo situation, it is also useful in understanding immune disorders in general. Thus, our proposed studies in TMC-NASA contract round II application benefit from data generated in this TMC-NASA contract round I study.

Long-term greenhouse gas measurements from aircraft

NASA Astrophysics Data System (ADS)

Karion, A.; Sweeney, C.; Wolter, S.; Newberger, T.; Chen, H.; Andrews, A.; Kofler, J.; Neff, D.; Tans, P.

2012-10-01

In March 2009 the NOAA/ESRL/GMD Carbon Cycle and Greenhouse Gases Group collaborated with the US Coast Guard (USCG) to establish the Alaska Coast Guard (ACG) sampling site, a unique addition to NOAA's atmospheric monitoring network. This collaboration takes advantage of USCG bi-weekly Arctic Domain Awareness (ADA) flights, conducted with Hercules C-130 aircraft from March to November each year. NOAA has installed window-replacement inlet plates on two USCG C-130 aircraft and deploys a pallet with NOAA instrumentation on each ADA flight. Flights typically last 8 h and cover a very large area, traveling from Kodiak, AK in the south up to Barrow, AK in the north, and making altitude profiles near the coast as well as in the interior. NOAA instrumentation on each flight includes: a flask sampling system, a continuous CO2/CH4/CO/H2O analyzer, a continuous ozone analyzer, and an ambient temperature and humidity sensor. GPS time and location from the aircraft's navigation system are also collected. Air samples collected in flight are analyzed at NOAA/ESRL for the major greenhouse gases and a variety of halocarbons and hydrocarbons that influence climate, stratospheric ozone, and air quality. Instruments on this aircraft are designed and deployed to be able to collect air samples and data autonomously, so that NOAA personnel visit the site only for installation at the beginning of each season. We present an assessment of the cavity ring-down spectroscopy (CRDS) CO2/CH4/CO/H2O analyzer performance operating on an aircraft over a three-year period. We describe the overall system for making accurate greenhouse gas measurements using a CRDS analyzer on an aircraft with minimal operator interaction. Short and long-term stability of the CRDS analyzer over a seven-month deployment period is better than 0.15 ppm, 2 ppb, and 5 ppb for CO2, CH4, CO respectively, considering differences of on-board reference tank measurements from a laboratory calibration performed prior to deployment. This stability is not affected by variation in pressure or temperature during flight. Biases and standard deviations of comparisons with flask samples suggest that atmospheric variability, flask-to-flask variability, and possible flask sampling biases may be driving biases in the comparison between flasks and in-situ CRDS measurements.

Metabolic Profiling of Geobacter sulfurreducens during Industrial Bioprocess Scale-Up.

PubMed

Muhamadali, Howbeer; Xu, Yun; Ellis, David I; Allwood, J William; Rattray, Nicholas J W; Correa, Elon; Alrabiah, Haitham; Lloyd, Jonathan R; Goodacre, Royston

2015-05-15

During the industrial scale-up of bioprocesses it is important to establish that the biological system has not changed significantly when moving from small laboratory-scale shake flasks or culturing bottles to an industrially relevant production level. Therefore, during upscaling of biomass production for a range of metal transformations, including the production of biogenic magnetite nanoparticles by Geobacter sulfurreducens, from 100-ml bench-scale to 5-liter fermentors, we applied Fourier transform infrared (FTIR) spectroscopy as a metabolic fingerprinting approach followed by the analysis of bacterial cell extracts by gas chromatography-mass spectrometry (GC-MS) for metabolic profiling. FTIR results clearly differentiated between the phenotypic changes associated with different growth phases as well as the two culturing conditions. Furthermore, the clustering patterns displayed by multivariate analysis were in agreement with the turbidimetric measurements, which displayed an extended lag phase for cells grown in a 5-liter bioreactor (24 h) compared to those grown in 100-ml serum bottles (6 h). GC-MS analysis of the cell extracts demonstrated an overall accumulation of fumarate during the lag phase under both culturing conditions, coinciding with the detected concentrations of oxaloacetate, pyruvate, nicotinamide, and glycerol-3-phosphate being at their lowest levels compared to other growth phases. These metabolites were overlaid onto a metabolic network of G. sulfurreducens, and taking into account the levels of these metabolites throughout the fermentation process, the limited availability of oxaloacetate and nicotinamide would seem to be the main metabolic bottleneck resulting from this scale-up process. Additional metabolite-feeding experiments were carried out to validate the above hypothesis. Nicotinamide supplementation (1 mM) did not display any significant effects on the lag phase of G. sulfurreducens cells grown in the 100-ml serum bottles. However, it significantly improved the growth behavior of cells grown in the 5-liter bioreactor by reducing the lag phase from 24 h to 6 h, while providing higher yield than in the 100-ml serum bottles. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Observations of Gas Emissions from Cascade Range Volcanoes (USA) using a Portable Real-Time Sensor Package and Evacuated Flasks

NASA Astrophysics Data System (ADS)

Kelly, P. J.; Werner, C. A.; Evans, W.; Ingebritsen, S.; Tucker, D.

2012-12-01

Degassing from most Cascade Range Volcanoes, USA, is characterized by low-temperature hydrothermal emissions. It is important to monitor these emissions as part of a comprehensive monitoring strategy yet access is often difficult and most features are sampled by the USGS only once per year at best. In an effort to increase the sampling frequency of major gas species and in preparation for building permanent, autonomous units, we built a portable sensor package capable of measuring H2O, CO2, SO2, and H2S in volcanic gas plumes. Here we compare results from the portable sensor package with gas analyses from direct samples obtained using a titanium tube and evacuated glass flasks collected at the same time. The sensor package is housed in a small, rugged case, weighs 5 kg, and includes sensors for measuring H2O (0-16 parts per thousand), CO2 (0-5000 ppmv), SO2 (0-100 ppm), and H2S (0-20 ppm) gases. Additional temperature and pressure sensors, a micro air pump, datalogger, and an internal battery are also incorporated. H2O and CO2 are measured using an infrared spectrometer (Licor 840) and sulfur-containing gases are measured using electrochemical sensors equipped with filters to mitigate cross-sensitivities. Data are collected at a 1 Hz sampling rate and can be recorded and displayed in real-time using a netbook computer or can be saved to the onboard datalogger. The data display includes timeseries of H2O, CO2, SO2, and H2S mixing ratios, the four-component bulk composition of the plume, and automated calculation of gas ratios commonly used in volcanic gas monitoring, such as H2O/CO2, CO2/SO2, and CO2/H2S . In the Cascade Range, the sensor package has been tested at Mt. Baker, Mt. St. Helens, Mt. Hood, and in Lassen Volcanic National Park. In each case, the instrument was placed 5 to 30 meters from the fumarole or fumarole field and emissions were sampled for 5 to 30 minutes. No SO2 was detected at any location. At Mt. Hood the sensor package yielded average CO2/H2S ratios from 10 to 16 in fumarole plumes versus flask CO2/H2S ratios (n = 2) of 13 and 16 on 9 July 2011, and on 28 July 2012 the sensor package yielded an average CO2/H2S ratio of 12 versus flask ratios (n = 2) of 13 (both sets of flask samples obtained in the Crater Rock area). At Mt. Baker, the sensor package yielded average CO2/H2S ratios from 19 to 22 whereas flask ratios (n = 3) were higher, from 25 to 32 (both fumarole-plume and flask samples obtained in the Sherman Crater area) on 22 July 2011. The mismatch falls slightly outside expected analytical uncertainty for the sensor package (about 20% relative for CO2/H2S ratios). However, flask samples collected in Sherman Crater in 2006 and 2007 (n = 5) yielded CO2/H2S ratios from 18 to 29, which nearly spans the range of observations in 2011. Therefore, one explanation for the small mismatch between the results of the sensor package and direct samples is that the sensor package measures bulk plume compositions that may integrate emissions from several chemically distinct fumaroles and the direct samples better represent the composition of discrete vents. Overall, the sensor package and evacuated flask data show good agreement and demonstrate that the real-time technique is a viable means for monitoring major volcanic gas species.

Navigation and vessel inspection circular No. 3-94. International maritime organization code for the safe carriage of irradiated nuclear fuel, plutonium and high-level radioactive wastes in flasks on board ships (IMO resolution a.748(18)). Final report

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

NONE

1994-05-26

The Circular calls the attention of Coast Guard field units, marine surveyors, shippers and carriers of nuclear materials to the International Maritime Organization (IMO) Code for the Safe Carriage of Irradiated Nuclear Fuel, Plutonium and High-Level Radioactive Wastes in Flasks on Board Ships (IMO Resolution A.748(18)).

Application of an Online-Biomass Sensor in an Optical Multisensory Platform Prototype for Growth Monitoring of Biotechnical Relevant Microorganism and Cell Lines in Single-Use Shake Flasks

PubMed Central

Ude, Christian; Schmidt-Hager, Jörg; Findeis, Michael; John, Gernot Thomas; Scheper, Thomas; Beutel, Sascha

2014-01-01

In the context of this work we evaluated a multisensory, noninvasive prototype platform for shake flask cultivations by monitoring three basic parameters (pH, pO2 and biomass). The focus lies on the evaluation of the biomass sensor based on backward light scattering. The application spectrum was expanded to four new organisms in addition to E. coli K12 and S. cerevisiae [1]. It could be shown that the sensor is appropriate for a wide range of standard microorganisms, e.g., L. zeae, K. pastoris, A. niger and CHO-K1. The biomass sensor signal could successfully be correlated and calibrated with well-known measurement methods like OD600, cell dry weight (CDW) and cell concentration. Logarithmic and Bleasdale-Nelder derived functions were adequate for data fitting. Measurements at low cell concentrations proved to be critical in terms of a high signal to noise ratio, but the integration of a custom made light shade in the shake flask improved these measurements significantly. This sensor based measurement method has a high potential to initiate a new generation of online bioprocess monitoring. Metabolic studies will particularly benefit from the multisensory data acquisition. The sensor is already used in labscale experiments for shake flask cultivations. PMID:25232914

Application of multivariate analysis and mass transfer principles for refinement of a 3-L bioreactor scale-down model--when shake flasks mimic 15,000-L bioreactors better.

PubMed

Ahuja, Sanjeev; Jain, Shilpa; Ram, Kripa

2015-01-01

Characterization of manufacturing processes is key to understanding the effects of process parameters on process performance and product quality. These studies are generally conducted using small-scale model systems. Because of the importance of the results derived from these studies, the small-scale model should be predictive of large scale. Typically, small-scale bioreactors, which are considered superior to shake flasks in simulating large-scale bioreactors, are used as the scale-down models for characterizing mammalian cell culture processes. In this article, we describe a case study where a cell culture unit operation in bioreactors using one-sided pH control and their satellites (small-scale runs conducted using the same post-inoculation cultures and nutrient feeds) in 3-L bioreactors and shake flasks indicated that shake flasks mimicked the large-scale performance better than 3-L bioreactors. We detail here how multivariate analysis was used to make the pertinent assessment and to generate the hypothesis for refining the existing 3-L scale-down model. Relevant statistical techniques such as principal component analysis, partial least square, orthogonal partial least square, and discriminant analysis were used to identify the outliers and to determine the discriminatory variables responsible for performance differences at different scales. The resulting analysis, in combination with mass transfer principles, led to the hypothesis that observed similarities between 15,000-L and shake flask runs, and differences between 15,000-L and 3-L runs, were due to pCO2 and pH values. This hypothesis was confirmed by changing the aeration strategy at 3-L scale. By reducing the initial sparge rate in 3-L bioreactor, process performance and product quality data moved closer to that of large scale. © 2015 American Institute of Chemical Engineers.

The Influence of Ecological Isolation on the Structural and Functional Stability of Complex Microbial Communities

NASA Technical Reports Server (NTRS)

Franklin, R. B.; Garland, J. L.; Mills, A. L.

2005-01-01

To help understand how the behavior of microorganisms and microbial communities in insular space habitats may differ from the behavior of these groups on Earth, long-term incubations (100+ days) were conducting using wastewater bioreactors (batch fed) designed to mimic "closed" and "open" ecological systems. The issue of immigration was considered, and the goal of the research was to determine whether the stability of microbial communities in space is reduced due to their prolonged isolation. Bioreactors were established by inoculating flasks of sterile synthetic wastewater with the microbial community obtained from a local treatment facility; each day, one-third of the medium in the flask was replaced with an equal volume of sterile artificial wastewater. Flasks were divided into two treatments: "closed" and "open" to recruitment of additional microorganisms. "Closed" flasks were maintained as described above, while the medium used to feed the "open" flasks was supplemented daily with a small amount of raw sewage (which provided a continuous source of new potential community members). Significant differences in microbial community structure and function developed in the two sets of communities, and the results suggest that the open community was more stable and better able to adjust to changing environmental conditions. Each community's resistance to environmental (temperature fluctuations) and biological stresses (starvation and invasion by an opportunistic pathogen Pseudomonas aeruginosa) was monitored. Experiments were also conducted to determine whether the effect of isolation changes depending on the microbial communities' initial diversity or composition; communities with a low(er) initial diversity were less stable. Overall, the results indicate that isolation will be an important factor influencing the activity of microbial communities on board spacecraft. A possible way of mitigating these effects would be to include communities with high initial diversity or to periodically re-inoculate the systems using diverse inocula transported from Earth.

Chemistries for Protection and Decontamination

DTIC Science & Technology

2008-03-01

t i o n Measurement of Oxygen Uptake, Supported Catalyst and CEES in Solvent-Free System TDA R e s e a r c h Tests with HD • At CUBRC (Buffalo...dual gas burettes for HD tests at CUBRC . The dual burette on the left was connected to a flask with the catalyst and HD (and to an empty flask as a...Hill, Zhen Luo, Daniel Hillesheim • ECBC: Larry Procell • CUBRC : Meg Stapleton, Rich Fitzpatrick • Battelle: John Ontiveros, Walter Miller • ARO

Influence of cultivating conditions on the alpha-galactosidase biosynthesis from a novel strain of Penicillium sp. in solid-state fermentation.

PubMed

Wang, C L; Li, D F; Lu, W Q; Wang, Y H; Lai, C H

2004-01-01

The work is intended to achieve optimum culture conditions of alpha-galactosidase production by a mutant strain Penicillium sp. in solid-state fermentation (SSF). Certain fermentation parameters involving incubation temperature, moisture content, initial pH value, inoculum and load size of medium, and incubation time were investigated separately. The optimal temperature and moisture level for alpha-galactosidase biosynthesis was found to be 30 degrees C and 50%, respectively. The range of pH 5.5-6.5 was favourable. About 40-50 g of medium in 250-ml flask and inoculum over 1.0 x 10(6) spores were suitable for enzyme production. Seventy-five hours of incubation was enough for maximum alpha-galactosidase production. Substrate as wheat bran supplemented with soyabean meal and beet pulp markedly improved the enzyme yield in trays. Under optimum culture conditions, the alpha-galactosidase activity from Penicillium sp. MAFIC-6 indicated 185.2 U g(-1) in tray of SSF. The process on alpha-galactosidase production in laboratory scale may have a potentiality of scaling-up.

Comparison of extraction techniques of robenidine from poultry feed samples.

PubMed

Wilga, Joanna; Wasik, Agata Kot-; Namieśnik, Jacek

2007-10-31

In this paper, effectiveness of six different commonly applied extraction techniques for the determination of robenidine in poultry feed has been compared. The sample preparation techniques included shaking, Soxhlet, Soxtec, ultrasonically assisted extraction, microwave - assisted extraction and accelerated solvent extraction. Comparison of these techniques was done with respect to the recovery extraction, temperature and time, reproducibility and solvent consumption. Every single extract was subjected to clean - up using aluminium oxide column (Pasteur pipette filled with 1g of aluminium oxide), from which robenidine was eluted with 10ml of methanol. The eluate from the clean-up column was collected in a volumetric flask, and finally it was analysed by HPLC-DAD-MS. In general, all extraction techniques were capable of isolating of robenidine from poultry feed, but the recovery obtained using modern extraction techniques was higher than that obtained using conventional techniques. In particular, accelerated solvent extraction was more superior to other techniques, which highlights the advantages of this sample preparation technique. However, in routine analysis, shaking and ultrasonically assisted extraction is still the preferred method for the solution of robenidine and other coccidiostatics.

Effect of flask vibration time on casting integrity, Surface Penetration and Coating Inclusion in lost foam casting of Al-Si Alloy

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

Karimian, Majid; Idris, M. H.; Ourdjini, A.

2011-01-17

The paper presents the result of an experimental investigation conducted on medium aluminum silicon alloy casting- LM6, using no-vacuum assisted lost foam casting process. The study is directed for establishing the relationship between the flask vibrations times developed for molded sample on the casting integrity, surface penetration and coating inclusion defects of the casting. Four different flask vibration times namely 180, 120, 90 and 60 sec. were investigated. The casting integrity was investigated in terms of fulfilling in all portions and edges. The surface penetration was measured using optical microscope whilst image analyzer was used to quantify the percentage ofmore » coating inclusion in the casting. The results show that vibration time has significant influence on the fulfilling as well as the internal integrity of the lost foam casting. It was found that the lower vibration time produced comparatively sound casing.« less

Shear stress enhances microcin B17 production in a rotating wall bioreactor, but ethanol stress does not

NASA Technical Reports Server (NTRS)

Gao, Q.; Fang, A.; Pierson, D. L.; Mishra, S. K.; Demain, A. L.

2001-01-01

Stress, including that caused by ethanol, has been shown to induce or promote secondary metabolism in a number of microbial systems. Rotating-wall bioreactors provide a low stress and simulated microgravity environment which, however, supports only poor production of microcin B17 by Escherichia coli ZK650, as compared to production in agitated flasks. We wondered whether the poor production is due to the low level of stress and whether increasing stress in the bioreactors would raise the amount of microcin B17 formed. We found that applying shear stress by addition of a single Teflon bead to a rotating wall bioreactor improved microcin B17 production. By contrast, addition of various concentrations of ethanol to such bioreactors (or to shaken flasks) failed to increase microcin B17 production. Ethanol stress merely decreased production and, at higher concentrations, inhibited growth. Interestingly, cells growing in the bioreactor were much more resistant to the growth-inhibitory and production-inhibitory effects of ethanol than cells growing in shaken flasks.

Atmospheric CO2 From Flask Air Samples at 10 Sites in the Scripps Institution of Oceanography (SIO) Air Sampling Network (1957 - 2001) (issued 2004)

DOE Data Explorer

Keeling, Charles D. [Univ. of California, San Diego, CA (United States). Scripps Inst. of Oceanography; Whorf, Timothy P. [Univ. of California, San Diego, CA (United States). Scripps Inst. of Oceanography; Blasing, T. J. [Carbon Dioxide Information Analysis Center (CDIAC), Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (USA); Jones, Sonja [Carbon Dioxide Information Analysis Center (CDIAC), Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (USA)

2004-09-01

The Carbon Dioxide Research Group, Scripps Institution of Oceanography, University of California, San Diego, has provided this data set, which includes long-term measurements of near-surface atmospheric CO2 concentrations at 10 locations spanning latitudes 82°N to 90°S. Most of the data are based on replicated (collected at the same time and place) flask samples taken at intervals of approximately one week to one month and subsequently subjected to infrared analysis. Periods of record begin in various years, ranging from 1957 (for the South Pole station) to 1985 (for Alert, Canada), and all flask data records except for Christmas Island and Baring Head, New Zealand extend through year 2001. Christmas Island data end with August, 2001 and Baring Head data end with October 2001. Weekly averages of continuous data from Mauna Loa Observatory, Hawaii, are available back to March 1958. Similar weekly averages are also available for La Jolla, California, from November 1972 to October 1975, and for the South Pole from June 1960 to October 1963. These long-term records of atmospheric CO2 concentration complement the continuous records made by SIO, and also complement the long term flask records of the Climate Monitoring and Diagnostics Laboratory of the National Oceanic and Atmospheric Administration. All these data are useful for characterizing seasonal and geographical variations in atmospheric CO2 over several years, and for assessing results of global carbon models.

Large-scale production of lentiviral vector in a closed system hollow fiber bioreactor

PubMed Central

Sheu, Jonathan; Beltzer, Jim; Fury, Brian; Wilczek, Katarzyna; Tobin, Steve; Falconer, Danny; Nolta, Jan; Bauer, Gerhard

2015-01-01

Lentiviral vectors are widely used in the field of gene therapy as an effective method for permanent gene delivery. While current methods of producing small scale vector batches for research purposes depend largely on culture flasks, the emergence and popularity of lentiviral vectors in translational, preclinical and clinical research has demanded their production on a much larger scale, a task that can be difficult to manage with the numbers of producer cell culture flasks required for large volumes of vector. To generate a large scale, partially closed system method for the manufacturing of clinical grade lentiviral vector suitable for the generation of induced pluripotent stem cells (iPSCs), we developed a method employing a hollow fiber bioreactor traditionally used for cell expansion. We have demonstrated the growth, transfection, and vector-producing capability of 293T producer cells in this system. Vector particle RNA titers after subsequent vector concentration yielded values comparable to lentiviral iPSC induction vector batches produced using traditional culture methods in 225 cm2 flasks (T225s) and in 10-layer cell factories (CF10s), while yielding a volume nearly 145 times larger than the yield from a T225 flask and nearly three times larger than the yield from a CF10. Employing a closed system hollow fiber bioreactor for vector production offers the possibility of manufacturing large quantities of gene therapy vector while minimizing reagent usage, equipment footprint, and open system manipulation. PMID:26151065

Potential hazards of toxic metals found in toothpastes commonly used in Nigeria.

PubMed

Orisakwe, Orish Ebere; Okolo, Kenneth Obinna; Igweze, Zelinjo Nkeiruka; Ajaezi, Godwin Chukwuebuka; Udowelle, Nnaemeka Arinze

2016-01-01

Toothpastes have multi-functional configurations as oral care products. They can however constitute a pos- sible source, amongst others, of toxic metal exposure in public health. Indeed, the public health impact of personal hygiene and consumer products is largely unknown. To determine the level of toxic metals (lead, cadmium, cobalt, chromium, nickel) in toothpastes available in Nigeria, (home produced and imported), and assess the potential risk to the people. The samples of toothpastes commonly used in Nigeria were tested. Using a market basket protocol thirty five different brands of toothpaste were used. Samples were digest by addition of 10 mL mixture of conc. nitric and hydrochloric acids (HCl:HNO(3), 3:1), followed by heating to dryness. 20 mL deionized water was added, stirred and filtered. The filtrate was made up in standard volumetric flask and lead, cadmium, chromium, cobalt and nickel concentrations were determined using the atomic absorption spectrophotometry 205A. The daily intake of metals and target hazard quotient (THQ) were then calculated. Pepsodent and Flodent had the highest levels of lead at respectively 23.575 and 18.092 mg/kg while Colgate Herbal had the highest nickel of 18.535 mg/kg. The daily intake estimates of all imported toothpaste samples were below the stated upper limits (UL). All target hazard quotients were also found to be below one. Although the UL, THQ and daily intake rates were all normal, the high levels of lead in some of the tooth- pastes an important concern to public health suggesting that pre-marketing safety studies of toothpastes may be worthwhile for the regulatory authorities.

A Simplified Extemporaneously Prepared Potassium Chloride Oral Solution.

PubMed

Tannous, Elias; Tal, Yana; Amarny, Kamal

2016-01-01

Although commercial preparations of oral potassium supplements are usually available, there are times when our Medical Center is faced with situations in which the oral solution of potassium chloride is not available. This solution is necessary for our pediatric outpatients who cannot swallow tablets and need an oral solution. Moreover, there are no studies available which describe an extemporaneously prepared potassium chloride oral solution on which we can rely for assigning a beyond-use date. The aim of this study was to formulate an extemporaneous pediatric oral solution of potassium chloride and to determine the physical and chemical stability of this preparation. We prepared 1 mMoL/mL by withdrawing 25 mL of potassium chloride 14.9%. Ora-Sweet SF was added to 50 mL in a metered flask. The solution was kept refrigerated (2°C to 8°C). Samples were withdrawn to measure potassium concentration, pH, and microbial overgrowth. The test was performed by our biochemical laboratory. The oral solution of potassium chloride 1 mMoL/mL stored at 2°C to 8°C maintained at least 91% of the initial concentration for 28 days. There were no notable changes in pH, and the solution remained physically stable with no visual microbial growth. The oral solution of potassium chloride 1 mMoL/mL prepared in Ora-Sweet and stored at 2°C to 8°C in amber glass bottles is expected to remain stable for 28 days. Copyright© by International Journal of Pharmaceutical Compounding, Inc.

Genetic Circuit Performance under Conditions Relevant for Industrial Bioreactors

PubMed Central

Moser, Felix; Broers, Nicolette J.; Hartmans, Sybe; Tamsir, Alvin; Kerkman, Richard; Roubos, Johannes A.; Bovenberg, Roel; Voigt, Christopher A.

2014-01-01

Synthetic genetic programs promise to enable novel applications in industrial processes. For such applications, the genetic circuits that compose programs will require fidelity in varying and complex environments. In this work, we report the performance of two synthetic circuits in Escherichia coli under industrially relevant conditions, including the selection of media, strain, and growth rate. We test and compare two transcriptional circuits: an AND and a NOR gate. In E. coli DH10B, the AND gate is inactive in minimal media; activity can be rescued by supplementing the media and transferring the gate into the industrial strain E. coli DS68637 where normal function is observed in minimal media. In contrast, the NOR gate is robust to media composition and functions similarly in both strains. The AND gate is evaluated at three stages of early scale-up: 100 ml shake-flask experiments, a 1 ml MTP microreactor, and a 10 L bioreactor. A reference plasmid that constitutively produces a GFP reporter is used to make comparisons of circuit performance across conditions. The AND gate function is quantitatively different at each scale. The output deteriorates late in fermentation after the shift from exponential to constant feed rates, which induces rapid resource depletion and changes in growth rate. In addition, one of the output states of the AND gate failed in the bioreactor, effectively making it only responsive to a single input. Finally, cells carrying the AND gate show considerably less accumulation of biomass. Overall, these results highlight challenges and suggest modified strategies for developing and characterizing genetic circuits that function reliably during fermentation. PMID:23656232

English Translation of Seven Papers on Thermophysical Properties

DTIC Science & Technology

1980-09-01

capillary constant (eq. 18) *- a = surface tension (eq. 18) g = acceleration of gravity (eq. 18) v = atomic volume at the melting point - . .. M-16...Accuracyoft/"m Ex*Ceperiment C 1941 r1143 oil 181-195 0.4610.01 Specific gravity flask 1946 [20 - t, Ar (up to 332-160 0.517 1.01 Specific gravity ...i 65 arm) flask, 0.52 1ś No1 P, 1 Kb 16 319T 400-1125 0.503 1.021 Specific gravity 1970 163) was - 242.-917 0.304 0.95 Hydrostatic sun-pension. -. 3

Atmospheric CH4 Concentrations from the Commonwealth Scientific and Industrial Research Organization (CSIRO) GASLAB Flask Sampling Network (1984 - 2001)

DOE Data Explorer

Steele, L. P. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Aspendale, Victoria, Australia; Krummel, P. B. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Aspendale, Victoria, Australia; Langenfelds, R. L. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Aspendale, Victoria, Australia

2003-01-01

The listed data were obtained from flask air samples returned to the CSIRO GASLAB for analysis. Typical sample storage times ranged from days to weeks for some sites (e.g., Cape Grim) to as much as one year for Macquarie Island and the Antarctic sites. Experiments carried out to test for any change in sample CH4 mixing ratio during storage have shown no drift to within detection limits over test periods of several months to years (Cooper et al., 1999).

Fluid Dynamics Appearing during Simulated Microgravity Using Random Positioning Machines

PubMed Central

Stern, Philip; Casartelli, Ernesto; Egli, Marcel

2017-01-01

Random Positioning Machines (RPMs) are widely used as tools to simulate microgravity on ground. They consist of two gimbal mounted frames, which constantly rotate biological samples around two perpendicular axes and thus distribute the Earth’s gravity vector in all directions over time. In recent years, the RPM is increasingly becoming appreciated as a laboratory instrument also in non-space-related research. For instance, it can be applied for the formation of scaffold-free spheroid cell clusters. The kinematic rotation of the RPM, however, does not only distribute the gravity vector in such a way that it averages to zero, but it also introduces local forces to the cell culture. These forces can be described by rigid body analysis. Although RPMs are commonly used in laboratories, the fluid motion in the cell culture flasks on the RPM and the possible effects of such on cells have not been examined until today; thus, such aspects have been widely neglected. In this study, we used a numerical approach to describe the fluid dynamic characteristic occurring inside a cell culture flask turning on an operating RPM. The simulations showed that the fluid motion within the cell culture flask never reached a steady state or neared a steady state condition. The fluid velocity depends on the rotational velocity of the RPM and is in the order of a few centimeters per second. The highest shear stresses are found along the flask walls; depending of the rotational velocity, they can reach up to a few 100 mPa. The shear stresses in the “bulk volume,” however, are always smaller, and their magnitude is in the order of 10 mPa. In conclusion, RPMs are highly appreciated as reliable tools in microgravity research. They have even started to become useful instruments in new research fields of mechanobiology. Depending on the experiment, the fluid dynamic on the RPM cannot be neglected and needs to be taken into consideration. The results presented in this study elucidate the fluid motion and provide insight into the convection and shear stresses that occur inside a cell culture flask during RPM experiments. PMID:28135286

Investigations on the Use of Multi-Species Flask Measurements for Sector-Specific Fossil Fuel CO2 Attribution to Aid Policymakers

NASA Astrophysics Data System (ADS)

Nathan, B.; Lauvaux, T.; Turnbull, J. C.; Sweeney, C.; Karion, A.; Richardson, S.; Miles, N.; Gurney, K. R.; Patarasuk, R.

2016-12-01

Part of the Indianapolis Flux (INFLUX) Experiment has, since 2010, involved recording atmospheric trace gas measurements using NOAA flask packages. The goal of these measurements is to better inform policymakers about the behaviors of greenhouse gas emissions in the Indianapolis urban environment. Radiocarbon dioxide (14CO2) measurements recorded from the flasks allow for delineation of the fossil-fuel carbon dioxide (CO2ff) signal from the total carbon dioxide (CO2) measurement. To give policymakers even more detailed information, we investigate whether the co-measured trace gases could be used as tracers for economic source sectors of CO2ff as predefined by the bottom-up data product Hestia. This is extensively tested using an Observation System Simulation Experiment (OSSE) combining both a top-down approach for all species—influence functions from the tower flask measurements—, and attempting to assign sources via spatial overlaps with the available bottom-up inventory CO2ff source sector definitions. A self-organizing map is implemented for the mathematical attribution of signals to sources, because it can compensate for nonlinear signals (i.e. tracer emissions that do not scale linearly with CO2ff emissions). It is determined that proper attribution is at least not feasible with such a complete lack of bottom-up spatial information about all non-CO2ff potential tracers. This unfeasibility is shown not to be resolved by a test of expanding the dataset with many more theoretical measurements than are realistically available. Here we alter the approach to include the missing prior information: bottom-up estimates of the emission fluxes for a suite of species. We develop these bottom-up emission fluxes from existing whole-city emission fluxes, species-specific source sector partitioning, and the spatial patterns from Hestia CO2ff source sectors. We validate the general approach using the whole-city species: CO2ff ratios derived from all tower flask measurements. Finally, using these tools, multi-species, sector-specific inversions are investigated in the Bayesian framework.

Atmospheric CO2 Records from Sites in the Atmospheric Environment Service Air Sampling Network (1975 and 1994)

DOE Data Explorer

Trivett, N. B.A. [Atmospheric Environment Service, Downsview, Ontario, Canada; Hudec, V. C. [Atmospheric Environment Service, Downsview, Ontario, Canada; Wong, C. S. [Marine Carbon Research Centre, Institute of Ocean Sciences, Sidney, British Columbia, Canada

1997-01-01

From the mid-1970s through the mid-1990s, air samples were collected for the purposes of monitoring atmospheric CO2 from four sites in the AES air sampling network. Air samples were collected approximately once per week, between 12:00 and 16:00 local time, in a pair of evacuated 2-L thick-wall borosilicate glass flasks. Samples were collected under preferred conditions of wind speed and direction (i.e., upwind of the main station and when winds are strong and steady). The flasks were evacuated to pressures of ~1 × 10-4 mbar or 0.01 Pa prior to being sent to the stations. The airwas not dried during sample collection. The flask data from Alert show an increase in the annual atmospheric CO2 concentration from 341.35 parts per million by volume (ppmv) in 1981 to 357.21 ppmv in 1991. For Cape St. James, Trivett and Higuchi (1989) reported that the mean annual rate of increase, obtained from the slope of a least-squares regression line through the annual averages, was 1.43 ppmv per year. In August 1992, the weather station at Cape St. James was automated; as a result, the flask sampling program was discontinued at this site. Estevan Point, on the West Coast of Vancouver Island, was chosen as a replacement station. Sampling at Estevan Point started in 1992; thus, the monthly and annual CO2record from Estevan Point is too short to show any long-term trends. The sampling site at Sable Island, off the coast of Nova Scotia, was established in 1975. The flask data from Sable Island show an increase in the annual atmospheric CO2 concentration from 334.49 parts per million by volume (ppmv) in 1977 (the first full year of data) to 356.02 ppmv in 1990. For Sable Island, Trivett and Higuchi (1989) reported that the mean annual rate of increase, obtained from the slope of a least-squares regression line through the annual averages, was 1.48 ppmv per year.

Pericellular oxygen monitoring with integrated sensor chips for reproducible cell culture experiments.

PubMed

Kieninger, J; Aravindalochanan, K; Sandvik, J A; Pettersen, E O; Urban, G A

2014-04-01

Here we present an application, in two tumour cell lines, based on the Sensing Cell Culture Flask system as a cell culture monitoring tool for pericellular oxygen sensing. T-47D (human breast cancer) and T98G (human brain cancer) cells were cultured either in atmospheric air or in a glove-box set at 4% oxygen, in both cases with 5% CO2 in the gas phase. Pericellular oxygen tension was measured with the help of an integrated sensor chip comprising oxygen sensor arrays. Obtained results illustrate variation of pericellular oxygen tension in attached cells covered by stagnant medium. Independent of incubation conditions, low pericellular oxygen concentration levels, usually associated with hypoxia, were found in dense cell cultures. Respiration alone brought pericellular oxygen concentration down to levels which could activate hypoxia-sensing regulatory processes in cultures believed to be aerobic. Cells in culture believed to experience conditions of mild hypoxia may, in reality, experience severe hypoxia. This would lead to incorrect assumptions and suggests that pericellular oxygen concentration readings are of great importance to obtain reproducible results when dealing with hypoxic and normoxic (aerobic) incubation conditions. The Sensing Cell Culture Flask system allows continuous monitoring of pericellular oxygen concentration with outstanding long-term stability and no need for recalibration during cell culture experiments. The sensor is integrated into the flask bottom, thus in direct contact with attached cells. No additional equipment needs to be inserted into the flask during culturing. Transparency of the electrochemical sensor chip allows optical inspection of cells attached on top of the sensor. © 2014 John Wiley & Sons Ltd.

Postural control under clinorotation in upside-down catfish, Synodontis nigriventris.

PubMed

Ohnishi, K; Takahashi, A; Koyama, M; Ohnishi, T

1996-12-01

The upside-down catfish Synodontis nigriventris has a unique habit of swimming and resting upside-down in free water. This behavior leads to the assumption that the catfish has a specific gravity information processing system. We examined the postural control behaviors in the catfish under clinorotation which is usually used for producing pseudo-microgravity. Synodontis nigriventris kept its body posture at a stable area of the rotated flask in which the catfish was kept, when it was clinorotated at the rate of 60 rpm. In contrast to Synodontis nigriventris, a related species, Corydoras paleatus, did not show such steady postural control. When the flask was rotated at a lower rate of 30 rpm or a higher rate of 100 rpm, Synodontis nigriventris as well as Corydoras paleatus showed a considerable disturbed control of body posture. In this condition, they were frequently rotated with the flask. These findings suggest that Synodontis nigriventris has a high ability to keep upside-down posture and the gravity sensation in this catfish is likely to contribute to its different postural control from that of many other fishes.

Flow characterization of a spinner flask for induced pluripotent stem cell culture application.

PubMed

Ismadi, Mohd-Zulhilmi; Gupta, Priyanka; Fouras, Andreas; Verma, Paul; Jadhav, Sameer; Bellare, Jayesh; Hourigan, Kerry

2014-01-01

We present detailed quantitative measurement analyses for flow in a spinner flask with spinning rates between 20 to 45 RPM, utilizing the optical velocimetry measurement technique of Particle Image Velocimetry (PIV). A partial section of the impeller was immersed in the working fluid to reduce the shear forces induced on the cells cultured on microcarriers. Higher rotational speeds improved the mixing effect in the medium at the expense of a higher shear environment. It was found that the mouse induced pluripotent stem (iPS) cells achieved the optimum number of cells over 7 days in 25 RPM suspension culture. This condition translates to 0.0984 Pa of maximum shear stress caused by the interaction of the fluid flow with the bottom surface. However, inverse cell growth was obtained at 28 RPM culture condition. Such a narrow margin demonstrated that mouse iPS cells cultured on microcarriers are very sensitive to mechanical forces. This study provides insight to biomechanical parameters, specifically the shear stress distribution, for a commercially available spinner flask over a wide range of Reynolds number.

Molecular Engineering of Liquid Crystalline Polymers by Living Polymerization. 10. Influence of Molecular Weight on the Phase Transitions of Poly(Omega-((4-Cyano-4’-Biphenyl)oxy)alkyl Vinyl Ethers)s with Nonyl and Decanyl Alkyl Groups

DTIC Science & Technology

1990-10-16

washed with concentrated sulfuric acid , then with water, dried over anhydrous magnesium sulfate, refluxed over calcium hydride and freshly distilled...oxide, filtered, and fractionally distilled under reduced pressure. Trifluoromethane sulfonic acid (triflic acid , 98%, Aldrich) w s distilled under...flask. Then the flask was filled with argon, cooled to 0°C and the methylene chloride, dimethyl sulfide and triflic acid were added via a syringe. The

Effect of a new tension system, used in acrylic resin flasking, on the dimensional stability of denture bases.

PubMed

Consani, Rafael Leonardo Xediek; Domitti, Saide Sarckis; Consani, Simonides

2002-09-01

The pressure of final closure may be released when the flask is removed from the mechanical or pneumatic press and placed in the spring clamp. This release in pressure may result in dimensional changes that distort the denture base. The purpose of this study was to investigate differences between the dimensional stability of standardized simulated denture bases processed by traditional moist heat-polymerization and those processed by use of a new tension system. A metal master die was fabricated to simulate an edentulous maxillary arch without irregularities in the alveolar ridge walls. A silicone mold of this metallic die was prepared, and 40 stone casts were formed from the mold with type III dental stone. The casts were randomly assigned to 4 test groups (A-D) of 10 specimens each. A uniform denture base pattern was made on each stone cast with a 1.5-mm thickness of base-plate wax, measured with a caliper. The patterns were invested for traditional hot water processing. A polymethyl methacrylate dough was prepared and packed for processing. The flasks in groups A and B were closed with the traditional pressure technique and placed in spring clamps after final closure. The flasks in groups C and D were pressed between the metallic plates of the new tension system after the final closure. The group A and C flasks were immediately immersed in the water processing unit at room temperature (25 degrees +/- 2 degrees C). The unit was programmed to raise the temperature to 74 degrees C over 1 hour, and then maintained the temperature at 74 degrees C for 8 hours. The group B and D flasks were bench stored at room temperature (25 degrees +/- 2 degrees C) for 6 hours and were then subjected to the same moist heat polymerization conditions as groups A and C. All processed dentures were bench cooled for 3 hours. After recovery from the flasks, the base-cast sets were transversally sectioned into 3 parts (corresponding to 3 zones): (1) distal of the canines, (2) mesial of the first molars, and (3) mesial of the posterior palate). These areas had been previously established and standardized by use of a pattern denture in the sawing device to determine the sections in each base-cast set. Base-cast gaps were measured at 5 predetermined points on each section with an optical micrometer that had a tolerance of 0.001 mm. Collected data were analyzed with analysis of variance and Tukey's test. Denture bases processed with the new tension system exhibited significantly better base adaptation than those processed with traditional acrylic resin packing. Immediately after polymerization (Groups A and C), mean dimensional change values were 0.213 +/- 0.055 mm for the traditional packing technique and 0.173 +/- 0.050 mm for new tension system. After delayed polymerization (Groups B and D), the values were 0.216 +/- 0.074 mm for the traditional packing technique and 0.164 +/- 0.032 mm for new tension system. With both techniques, dimensional changes in the posterior palatal zone were greater (conventional = 0.286 +/- 0.038 mm; new system = 0.214 +/- 0.024 mm) than those elsewhere on the base-cast set. Within the limitations of this study, the new tension packing system was associated with decreased dimensional changes in the simulated maxillary denture bases processed with heat-polymerization.

Simulated conditions of microgravity suppress progesterone production by luteal cells of the pregnant rat

NASA Technical Reports Server (NTRS)

Bhat, G. K.; Yang, H.; Sridaran, R.

2001-01-01

The purpose of this study was to assess whether simulated conditions of microgravity induce changes in the production of progesterone by luteal cells of the pregnant rat ovary using an in vitro model system. The microgravity environment was simulated using either a high aspect ratio vessel (HARV) bioreactor with free fall or a clinostat without free fall of cells. A mixed population of luteal cells isolated from the corpora lutea of day 8 pregnant rats was attached to cytodex microcarrier beads (cytodex 3). These anchorage dependent cells were placed in equal numbers in the HARV or a spinner flask control vessel in culture conditions. It was found that HARV significantly reduced the daily production of progesterone from day 1 through day 8 compared to controls. Scanning electron microscopy showed that cells attached to the microcarrier beads throughout the duration of the experiment in both types of culture vessels. Cells cultured in chamber slide flasks and placed in a clinostat yielded similar results when compared to those in the HARV. Also, when they were stained by Oil Red-O for lipid droplets, the clinostat flasks showed a larger number of stained cells compared to control flasks at 48 h. Further, the relative amount of Oil Red-O staining per milligram of protein was found to be higher in the clinostat than in the control cells at 48 h. It is speculated that the increase in the level of lipid content in cells subjected to simulated conditions of microgravity may be due to a disruption in cholesterol transport and/or lesions in the steroidogenic pathway leading to a fall in the synthesis of progesterone. Additionally, the fall in progesterone in simulated conditions of microgravity could be due to apoptosis of luteal cells.

Influence of Light Intensity on Growth and Pigment Production by Monascus ruber in Submerged Fermentation.

PubMed

Bühler, Rose Marie Meinicke; Müller, Bruna Luíse; Moritz, Denise Esteves; Vendruscolo, Francielo; de Oliveira, Debora; Ninow, Jorge Luiz

2015-07-01

To reduce environmental problems caused by glycerine accumulation and to make the production of biodiesel more profitable, crude glycerin without treatment was used as substrate for obtaining higher value-added bioproducts. Monascus ruber is a filamentous fungus that produces pigments, particularly red ones, which are used for coloring foods (rice wine and meat products). The interest in developing pigments from natural sources is increasing due to the restriction of using synthetic dyes. The effects of temperature, pH, microorganism morphology, aeration, nitrogen source, and substrates have been studied in the cultivation of M. ruber. In this work, it was observed that light intensity is also an important factor that should be considered for understanding the metabolism of the fungus. In M. ruber cultivation, inhibition of growth and pigment production was observed in Petri dishes and blaffed flasks exposed to direct illumination. Growth and pigment production were higher in Petri dishes and flasks exposed to red light and in the absence of light. Radial growth rate of M. ruber in plates in darkness was 1.50 mm day(-1) and in plates exposed to direct illumination was 0.59 mm day(-1). Maximum production of red pigments (8.32 UA) and biomass (8.82 g L(-1)) were obtained in baffled flasks covered with red film and 7.17 UA of red pigments, and 7.40 g L(-1) of biomass was obtained in flasks incubated in darkness. Under conditions of 1248 lux of luminance, the maximum pigment production was 4.48 UA, with production of 6.94 g L(-1) of biomass, indicating that the fungus has photoreceptors which influence the physiological responses.

Three-dimensional dynamic fabrication of engineered cartilage based on chitosan/gelatin hybrid hydrogel scaffold in a spinner flask with a special designed steel frame.

PubMed

Song, Kedong; Li, Liying; Li, Wenfang; Zhu, Yanxia; Jiao, Zeren; Lim, Mayasari; Fang, Meiyun; Shi, Fangxin; Wang, Ling; Liu, Tianqing

2015-10-01

Cartilage transplantation using in vitro tissue engineered cartilage is considered a promising treatment for articular cartilage defects. In this study, we assessed the advantages of adipose derived stem cells (ADSCs) combined with chitosan/gelatin hybrid hydrogel scaffolds, which acted as a cartilage biomimetic scaffold, to fabricate a tissue engineered cartilage dynamically in vitro and compared this with traditional static culture. Physical properties of the hydrogel scaffolds were evaluated and ADSCs were inoculated into the hydrogel at a density of 1×10(7) cells/mL and cultured in a spinner flask with a special designed steel framework and feed with chondrogenic inductive media for two weeks. The results showed that the average pore size, porosity, swelling rate and elasticity modulus of hybrid scaffolds with good biocompatibility were 118.25±19.51 μm, 82.60±2.34%, 361.28±0.47% and 61.2±0.16 kPa, respectively. ADSCs grew well in chitosan/gelatin hybrid scaffold and successfully differentiated into chondrocytes, showing that the scaffolds were suitable for tissue engineering applications in cartilage regeneration. Induced cells cultivated in a dynamic spinner flask with a special designed steel frame expressed more proteoglycans and the cell distribution was much more uniform with the scaffold being filled mostly with extracellular matrix produced by cells. A spinner flask with framework promoted proliferation and chondrogenic differentiation of ADSCs within chitosan/gelatin hybrid scaffolds and accelerated dynamic fabrication of cell-hydrogel constructs, which could be a selective and good method to construct tissue engineered cartilage in vitro. Copyright © 2015 Elsevier B.V. All rights reserved.

Evaluating maturation and genetic modification of human dendritic cells in a new polyolefin cell culture bag system.

PubMed

Macke, Lars; Garritsen, Henk S P; Meyring, Wilhelm; Hannig, Horst; Pägelow, Ute; Wörmann, Bernhard; Piechaczek, Christoph; Geffers, Robert; Rohde, Manfred; Lindenmaier, Werner; Dittmar, Kurt E J

2010-04-01

Dendritic cells (DCs) are applied worldwide in several clinical studies of immune therapy of malignancies, autoimmune diseases, and transplantations. Most legislative bodies are demanding high standards for cultivation and transduction of cells. Closed-cell cultivating systems like cell culture bags would simplify and greatly improve the ability to reach these cultivation standards. We investigated if a new polyolefin cell culture bag enables maturation and adenoviral modification of human DCs in a closed system and compare the results with standard polystyrene flasks. Mononuclear cells were isolated from HLA-A*0201-positive blood donors by leukapheresis. A commercially available separation system (CliniMACS, Miltenyi Biotec) was used to isolate monocytes by positive selection using CD14-specific immunomagnetic beads. The essentially homogenous starting cell population was cultivated in the presence of granulocyte-macrophage-colony-stimulating factor and interleukin-4 in a closed-bag system in parallel to the standard flask cultivation system. Genetic modification was performed on Day 4. After induction of maturation on Day 5, mature DCs could be harvested and cryopreserved on Day 7. During the cultivation period comparative quality control was performed using flow cytometry, gene expression profiling, and functional assays. Both flasks and bags generated mature genetically modified DCs in similar yields. Surface membrane markers, expression profiles, and functional testing results were comparable. The use of a closed-bag system facilitated clinical applicability of genetically modified DCs. The polyolefin bag-based culture system yields DCs qualitatively and quantitatively comparable to the standard flask preparation. All steps including cryopreservation can be performed in a closed system facilitating standardized, safe, and reproducible preparation of therapeutic cells.

Process optimization involving critical evaluation of oxygen transfer, oxygen uptake and nitrogen limitation for enhanced biomass and lipid production by oleaginous yeast for biofuel application.

PubMed

Chopra, Jayita; Sen, Ramkrishna

2018-04-20

Lipid accumulation in oleaginous yeast is generally induced by nitrogen starvation, while oxygen saturation can influence biomass growth. Systematic shake flask studies that help in identifying the right nitrogen source and relate its uptake kinetics to lipid biosynthesis under varying oxygen saturation conditions are very essential for addressing the bioprocessing-related issues, which are envisaged to occur in the fermenter scale production. In the present study, lipid bioaccumulation by P. guilliermondii at varying C:N ratios and oxygen transfer conditions (assessed in terms of k L a) was investigated in shake flasks using a pre-optimized N-source and a two-stage inoculum formulated in a hybrid medium. A maximum lipid concentration of 10.8 ± 0.5 g L -1 was obtained in shake flask study at the optimal condition with an initial C:N and k L a of 60:1 and 0.6 min -1 , respectively, at a biomass specific growth rate of 0.11 h -1 . Translating these optimal shake flask conditions to a 3.7 L stirred tank reactor resulted in biomass and lipid concentrations of 16.74 ± 0.8 and 8 ± 0.4 g L -1 . The fatty acid methyl ester (FAME) profile of lipids obtained by gas chromatography was found to be suitable for biodiesel application. We strongly believe that the rationalistic approach-based design of experiments adopted in the study would help in achieving high cell density with improved lipid accumulation and also minimize the efforts towards process optimization during bioreactor level operations, consequently reducing the research and development-associated costs.

Hydrogen isotope correction for laser instrument measurement bias at low water vapor concentration using conventional isotope analyses: application to measurements from Mauna Loa Observatory, Hawaii.

PubMed

Johnson, L R; Sharp, Z D; Galewsky, J; Strong, M; Van Pelt, A D; Dong, F; Noone, D

2011-03-15

The hydrogen and oxygen isotope ratios of water vapor can be measured with commercially available laser spectroscopy analyzers in real time. Operation of the laser systems in relatively dry air is difficult because measurements are non-linear as a function of humidity at low water concentrations. Here we use field-based sampling coupled with traditional mass spectrometry techniques for assessing linearity and calibrating laser spectroscopy systems at low water vapor concentrations. Air samples are collected in an evacuated 2 L glass flask and the water is separated from the non-condensable gases cryogenically. Approximately 2 µL of water are reduced to H(2) gas and measured on an isotope ratio mass spectrometer. In a field experiment at the Mauna Loa Observatory (MLO), we ran Picarro and Los Gatos Research (LGR) laser analyzers for a period of 25 days in addition to periodic sample collection in evacuated flasks. When the two laser systems are corrected to the flask data, they are strongly coincident over the entire 25 days. The δ(2)H values were found to change by over 200‰ over 2.5 min as the boundary layer elevation changed relative to MLO. The δ(2)H values ranged from -106 to -332‰, and the δ(18)O values (uncorrected) ranged from -12 to -50‰. Raw data from laser analyzers in environments with low water vapor concentrations can be normalized to the international V-SMOW scale by calibration to the flask data measured conventionally. Bias correction is especially critical for the accurate determination of deuterium excess in dry air. Copyright © 2011 John Wiley & Sons, Ltd.

Establishing Regular Measurements of Halocarbons at Taunus Observatory

NASA Astrophysics Data System (ADS)

Schuck, Tanja; Lefrancois, Fides; Gallmann, Franziska; Engel, Andreas

2017-04-01

In late 2013 an ongoing whole air flask collection program has been started at the Taunus Observatory (TO) in central Germany. Being a rural site in close vicinity to the densely populated Rhein-Main area with the city of Frankfurt, Taunus Observatory allows to assess local and regional emissions but owed to its altitude of 825m also regularly experiences background conditions. With its large caption area halocarbon measurements at the site have the potential to improve the data base for estimation of regional and total European halogenated greenhouse gas emissions. At current, flask samples are collected weekly for analysis using a GC-MS system at Frankfurt University employing a quadrupole as well as a time-of-flight (TOF) mass spectrometer. The TOF instrument yields full scan mass information and allows for retrospective analysis of so far undetected non-target species. For quality assurance additional samples are collected approximately bi-weekly at the Mace Head Atmospheric Research Station (MHD) analyzed in Frankfurt following the same measurement procedure. Thus the TO time series can be linked to both, the in-situ AGAGE measurements and the NOAA flask sampling program at MHD. In 2017 it is planned to supplement the current flask sampling by employing an in-situ GC-MS system at the site, thus increasing the measurement frequency. We will present the timeseries of selected halocarbons recorded at Taunus Observatory. While there is good agreement of baseline mixing ratios between TO and MHD, measurements at TO are regularly influenced by elevated halocarbon mixing ratios. An analysis of HYSPLIT trajectories for the existing time series revealed significant differences in halocarbon mixing ranges depending on air mass origin.

Effect of light with different wavelengths on Nostoc flagelliforme cells in liquid culture.

PubMed

Dai, Yu-Jie; Li, Jing; Wei, Shu-Mei; Chen, Nan; Xiao, Yu-Peng; Tan, Zhi-Lei; Jia, Shi-Ru; Yuan, Nan-Nan; Tan, Ning; Song, Yi-Jie

2013-04-01

The effects of lights with different wavelengths on the growth and the yield of extracellular polysaccharides of Nostoc flagelliforme cells were investigated in a liquid cultivation. N. flagelliforme cells were cultured for 16 days in 500 ml conical flasks containing BG11 culture medium under 27 micromol·m-2·s-1 of light intensity and 25 degrees C on a rotary shaker (140 rpm). The chlorophyll a, phycocyanin, allophycocyanin, and phycoerythrin contents in N. flagelliforme cells under the lights of different wavelengths were also measured. It was found that the cell biomass and the yield of polysaccharide changed with different wavelengths of light. The biomass and the yield of extracellular polysaccharides under the red or violet light were higher than those under other light colors. Chlorophyll a, phycocyanin, and allophycocyanin are the main pigments in N. flagelliforme cells. The results showed that N. flagelliforme, like other cyanobacteria, has the ability of adjusting the contents and relative ratio of its pigments with the light quality. As a conclusion, N. flagelliforme cells favor red and violet lights and perform the complementary chromatic adaptation ability to acclimate to the changes of the light quality in the environment.

Comparison of different rubber stoppers; the effect of sterilization on the number of particles released.

PubMed

Mannermaa, J P; Muttonen, E; Yliruusi, J; Juppo, A

1992-01-01

The effect of sterilization on the number of particles released from five different types of rubber stoppers, as well as on their surface roughness and elemental composition before and after sterilization is described. The stoppers were immersed in 200 ml of 0.9% sodium chloride solution in conical flasks. The number of particles released into the sodium chloride solution was measured by Coulter Counter. The surface roughness and the elemental composition of the stoppers were determined by SEM/EDX. All measurements were made both before and after sterilization at 121 degrees C to F0 15 mins. The number of particles released from a stopper during sterilization varies considerably between different stoppers and even between different batches of the same stopper. The only non-siliconized stopper in this study performed well. The absence of surface siliconization may have contributed to this performance. The scanning electron micrographs revealed well the differences in the surface roughness of the stoppers. The sterilization generally increases the surface roughness of the samples. The x-ray microanalysis revealed that the elemental composition of the stoppers may vary not only between different types of stoppers but also between different batches of the same stopper.

Response of sago pondweed, a submerged aquatic macrophyte, to herbicides in three laboratory culture systems

USGS Publications Warehouse

Fleming, W.J.; Ailstock, M.S.; Momot, J.J.; Norman, C.M.; Gorsuch, Joseph W.; Lower, William R.; Wang, Wun-cheng; Lewis, M.A.

1991-01-01

The phytotoxicity of atrazine, paraquat, glyphosate, and alachlor to sago pondweed (Potamogeton pectinatus), a submerged aquatic macrophyte, was tested under three types of laboratory culture conditions. In each case, tests were conducted in static systems, the test period was four weeks, and herbicide exposure was chronic, resulting from a single addition of herbicide to the test vessels at the beginning of the test period. The three sets of test conditions employed were(1) axenic cultures in 125-mL flasks containing a nutrient media and sucrose; (2) a microcosm system employing 18.9-L buckets containing a sand, shell, and peat substrate; and (3) an algae-free system employing O.95-L jars containing reconstituted freshwater and a nutrient agar substrate. The primary variable measured was biomass production. Plants grew well in all three test systems, with biomass of untreated plants increasing by a factor of about 5 to 6.5 during the four-week test period. Biomass production in response to herbicide exposure differed significantly among culture systems, which demonstrates the need for a standardized testing protocol for evaluating the effects of toxics on submerged aquatic plants.

Semi-continuous production of high-activity pectinases by immobilized Rhizopus oryzae using tobacco wastewater as substrate and their utilization in the hydrolysis of pectin-containing lignocellulosic biomass at high solid content.

PubMed

Zheng, Yu-Xi; Wang, Yuan-Liang; Pan, Jun; Zhang, Jian-Rong; Dai, Ya; Chen, Kun-Yan

2017-10-01

In this study, highly reactive endo- and exo-polygalacturonases (PGs) were produced from the tobacco industry wastewater using immobilized Rhizopus oryzae. Compared with free cells, immobilized cells increased enzyme activity 2.8-fold and reduced production time to 24h by shake-flask production. Moreover, the immobilized cells enabled the semi-continuous production of enzymes through repeated-batch mode for seven consecutive cycles in a scale-up bioreactor. During the first five cycles, the average endo-PG and exo-PG activities reached 307.5 and 242.6U/ml, respectively. The addition of crude enzyme for the hydrolysis of pectin-containing lignocellulosic biomass under high-gravity conditions increased glucose release 4.2-fold (115.4 vs. 29.0g/L), compared with hydrolysis using cellulase alone. This process achieves the efficient production of pectin-degrading enzymes, provides a cost-effective method for tobacco wastewater treatment, and offers the possibility to obtain fermentable sugars with high-titer from pectin-containing lignocellulosic biomass, which has important potential for the commercial production of bio-fuels. Copyright © 2017 Elsevier Ltd. All rights reserved.

[Production of infectious units of Isaria fumosorosea (Hypocreales: Cordycipitaceae) from different indigenous isolates of northeastern Mexico using 3 propagation strategies].

PubMed

Gandarilla-Pacheco, Fatima L; Morales-Ramos, Lilia H; Pereyra-Alférez, Benito; Elías-Santos, Myriam; Quintero-Zapata, Isela

The aim of this study was to evaluate the production of blastospores and conidia of different native isolates and a strain of Isaria fumosorosea using different propagation techniques. Two liquid culture media of casamino acids and peptone as nitrogen sources and glucose as carbon source for both media cultures were respectively used in the production of blastospores, while for the production of conidia, the fungi were grown in potato dextrose agar; from these cultures, solutions of conidia to a concentration of 1×10 6 per milliliter were prepared to inoculate flasks with Sabouraud dextrose broth for the liquid phase of the biphasic culture, also known as preculture. Subsequently, rice grain bags were inoculated with the preculture and the conidia solutions, which were incubated for 14 days for solid fermentation and biphasic culture, respectively. The HIB-23 isolate recorded a concentration of 4.90×10 8 blastospores/ml in the casamino acid medium, while a concentration of 2.15×10 8 blastospores/ml was obtained in the peptone collagen medium. For the Pfr-612 strain, the conidia production in solid-state fermentation was 1.58×10 9 conidia/g, and for HIB-30 in the biphasic culture of 9.00×10 6 conidia/g. Solid-state fermentation proved to be the most effective method with an average of 1.09×10 9 conidia/g, whereas the biphasic culture was the least effective method with 2.76×10 6 conidia/g; no significant difference was reported for the submerged production media. Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

Short communication: Eicosatrienoic acid and docosatrienoic acid do not promote vaccenic acid accumulation in mixed ruminal cultures.

PubMed

AbuGhazaleh, A A; Holmes, L D; Jacobson, B N; Kalscheur, K F

2006-11-01

Previous research found that docosahexaenoic acid (C22:6n-3) was a component of fish oil that promotes trans-C18:1 accumulation in ruminal cultures when incubated with linoleic acid. The objective of this study was to determine if eicosatrienoic acid (C20:3n-3) and docosatrienoic acid (C22:3n-3), n-3 fatty acids in fish oil, promote accumulation of trans-C18:1, vaccenic acid (VA) in particular, using cultures of mixed ruminal microorganisms. Treatments consisted of control, control plus 5 mg of C20:3n-3 (ETA), control plus 5 mg of C22:3n-3 (DTA), control plus 15 mg of linoleic acid (LA), control plus 5 mg of C20:3n-3 and 15 mg of linoleic acid (ETALA), and control plus 5 mg of C22:3n-3 and 15 mg of linoleic acid (DTALA). Treatments were incubated in triplicate in 125-mL flasks, and 5 mL of culture contents was taken at 0 and 24 h for fatty acid analysis by gas-liquid chromatography. After 24 h of incubation, the concentrations of trans-C18:1 (0.87, 0.88, and 0.99 mg/culture), and VA (0.52, 0.56, and 0.62 mg/culture) were similar for the control, ETA, and DTA cultures, respectively. The concentrations of trans-C18:1 (5.51, 5.41, and 5.36 mg/culture), and VA (4.78, 4.62, and 4.59 mg/culture) were also similar between LA, ETALA, and DTALA cultures, respectively. These data suggest that C20:3n-3 and C22:3n-3 are not the active components in fish oil that promote VA accumulation when incubated with linoleic acid.

ARM Carbon Cycle Gases Flasks at SGP Site

DOE Data Explorer

Biraud, Sebastien

2013-03-26

Data from flasks are sampled at the Atmospheric Radiation Measurement Program ARM, Southern Great Plains Site and analyzed by the National Oceanic and Atmospheric Administration NOAA, Earth System Research Laboratory ESRL. The SGP site is included in the NOAA Cooperative Global Air Sampling Network. The surface samples are collected from a 60 m tower at the ARM SGP Central Facility, usually once per week in the afternoon. The aircraft samples are collected approximately weekly from a chartered aircraft, and the collection flight path is centered over the tower where the surface samples are collected. The samples are collected by the ARM and LBNL Carbon Project.

Measurements of Atmospheric Methane and 13C/12C of Atmospheric Methane from Flask Air Samples (1999)

DOE Data Explorer

Quay, Paul [School of Oceanography, University of Washington; Stutsman, Johnny [School of Oceanography, University of Washington

1999-01-01

This database offers precise measurements of atmospheric methane and 13C/12C in atmospheric methane from flask air samples collected at eight sites worldwide and aboard NOAA cruises in the Pacific Ocean. The eight sites include Olympic Peninsula, Washington; Cape Grim, Tasmania; Fraserdale, Ontario; Marshall Islands; Baring Head, New Zealand; Mauna Loa, Hawaii; Point Barrow, Alaska; and American Samoa. The measurements span the period 1988 to mid-1996. These data are useful for global methane budget analyses and for determining the atmospheric isotopic composition of methane. All isotopic measurements have been corrected for standard drift.

Spectral lights trigger biomass accumulation and production of antioxidant secondary metabolites in adventitious root cultures of Stevia rebaudiana (Bert.).

PubMed

Idrees, Muhammad; Sania, Bibi; Hafsa, Bibi; Kumari, Sana; Khan, Haji; Fazal, Hina; Ahmad, Ishfaq; Akbar, Fazal; Ahmad, Naveed; Ali, Sadeeq; Ahmad, Nisar

2018-05-30

Stevia rebaudiana (S. rebaudiana) is the most important therapeutic plant species and has been accepted as such worldwide. It has a tendency to accumulate steviol glycosides, which are 300 times sweeter than marketable sugar. Recently, diabetic patients commonly use this plant as a sugar substitute for sweet taste. In the present study, the effects of different spectral lights were investigated on biomass accumulation and production of secondary metabolites in adventitious root cultures of S. rebaudiana. For callus development, leaf explants were excised from seed-derived plantlets and inoculated on a Murashige and Skoog (MS) medium containing the combination of 2,4-dichlorophenoxy acetic acid (2, 4-D, 2.0mg/l) and 6-benzyladenine (BA, 2.0mg/l), while 0.5mg/l naphthalene acetic acid (NAA) was used for adventitious root culture. Adventitious root cultures were exposed to different spectral lights (blue, green, violet, red and yellow) for a 30-day period. White light was used as control. The growth kinetics was studied for 30days with 3-day intervals. In this study, the violet light showed the maximum accumulation of fresh biomass (2.495g/flask) as compared to control (1.63g/flask), while red light showed growth inhibition (1.025g/flask) as compared to control. The blue light enhanced the highest accumulation of phenolic content (TPC; 6.56mg GAE/g DW), total phenolic production (TPP; 101mg/flask) as compared to control (5.44mg GAE/g DW; 82.2mg GAE/g DW), and exhibited a strong correlation with dry biomass. Blue light also improved the accumulation of total flavonoid content (TFC; 4.33mg RE/g DW) and total flavonoid production (TFP; 65mg/flask) as compared to control. The violet light showed the highest DPPH inhibition (79.72%), while the lowest antioxidant activity was observed for control roots (73.81%). Hence, we concluded that the application of spectral lights is an auspicious strategy for the enhancement of the required antioxidant secondary metabolites in adventitious root cultures of S. rebaudiana and of other medicinal plants. Copyright © 2018 Académie des sciences. Published by Elsevier Masson SAS. All rights reserved.

Orbital experiment ``Gravisensor'': phototropic reactions of the moss Physcomitrella patens to different types of LED lighting.

NASA Astrophysics Data System (ADS)

Nikitin, Vladimir; Berkovich, Yuliy A.; Skripnikov, Alexander; Zyablova, Natalya; Mukhoyan, Makar; Emelianov, Grigory

The experiment was conducted on Russian Biological Satelite Bion-M #1 19.04-19.05 2013. Five transparent plastic cultural flasks were placed in five light isolated sections of Biocont-B2 cylindrical container with inner diameter of 120 mm and height of 230 mm. In four sections the flasks could be illuminated by top or side LED with wavelength of 458 nm, 630 nm, 730 nm, and white (color temperature 5000° K, peaks 453, 559 nm). Photon flux in each variant was 15 umol/(m2c). In the fifth section the flask with the shoots was in conditions of constant dark. Each section was equipped with its own video camera module. Cameras, video recorder and lighting were managed by micro controller. 12 days before launch, 5 tips of the moss shoots were explanted at each of the five flasks on the agar medium with nutrient components and were cultivated under white fluorescent lamps at 12 hour photo period till the launch. After entering the orbit and during next 14 days of flight top LEDs were turned on above the flasks. Then for the following 14 days of flight the side LEDs of similar wavelength were turned on. The moss gametophores were cultivated at 12-h photoperiod. During the experiment on an hourly basis a video recording of the moss was performed. Similar equipment was used for ground control. After the experiment video files were used to produce separate time-lapse films for each flask using AviSynth program. In flight the shoots demonstrated the maximum growth speed with far red lighting and slower speed with white lighting. With blue and red lighting after switching to side light stimuli the growth of shoots almost stopped. In the dark the shoots continued to grow until the 13 day after launch of the satellite, then their growth stopped. In ground control the relation of growth rate with various LEDs remained basically the same, with the exception of side blue lighting, where the shoots demonstrated considerable vertical growth. In flight the angle of inclination towards the light source was maximal (about 90º) with white lighting, and somewhat smaller with 730 nm. Under red and blue light the angle of phototropic inclination was difficult to measure due to poor growth of the shoots.In ground control the growth rate under blue light was several times higher, than in flight and final degree of inclination of the shoot tip came to about 10º. In ground control under side red lighting the growth was weak, while demonstrating a pronounced phototropic bend of 90º. In ground control in the dark a vertical growth of one shoot was observed with the rate somewhat larger, than in flight variant. Data on the dynamics of inclination of experimental and control plants are presented. The acquired data will be used to analyse the mechanisms of phototropic growth changes of moss shoots.

Persistence of Mycobacterium avium subsp. paratuberculosis and Other Zoonotic Pathogens during Simulated Composting, Manure Packing, and Liquid Storage of Dairy Manure

PubMed Central

Grewal, Sukhbir K.; Rajeev, Sreekumari; Sreevatsan, Srinand; Michel, Frederick C.

2006-01-01

Livestock manures contain numerous microorganisms which can infect humans and/or animals, such as Escherichia coli O157:H7, Listeria monocytogenes, Salmonella spp., and Mycobacterium avium subsp. paratuberculosis (Mycobacterium paratuberculosis). The effects of commonly used manure treatments on the persistence of these pathogens have rarely been compared. The objective of this study was to compare the persistence of artificially inoculated M. paratuberculosis, as well as other naturally occurring pathogens, during the treatment of dairy manure under conditions that simulate three commonly used manure management methods: thermophilic composting at 55°C, manure packing at 25°C (or low-temperature composting), and liquid lagoon storage. Straw and sawdust amendments used for composting and packing were also compared. Manure was obtained from a large Ohio free-stall dairy herd and was inoculated with M. paratuberculosis at 106 CFU/g in the final mixes. For compost and pack treatments, this manure was amended with sawdust or straw to provide an optimal moisture content (60%) for composting for 56 days. To simulate liquid storage, water was added to the manure (to simulate liquid flushing and storage) and the slurry was placed in triplicate covered 4-liter Erlenmeyer flasks, incubated under ambient conditions for 175 days. The treatments were sampled on days 0, 3, 7, 14, 28, and 56 for the detection of pathogens. The persistence of M. paratuberculosis was also assessed by a PCR hybridization assay. After 56 days of composting, from 45 to 60% of the carbon in the compost treatments was converted to CO2, while no significant change in carbon content was observed in the liquid slurry. Escherichia coli, Salmonella, and Listeria were all detected in the manure and all of the treatments on day 0. After 3 days of composting at 55°C, none of these organisms were detectable. In liquid manure and pack treatments, some of these microorganisms were detectable up to 28 days. M. paratuberculosis was detected by standard culture only on day 0 in all the treatments, but was undetectable in any treatment at 3 and 7 days. On days 14, 28, and 56, M. paratuberculosis was detected in the liquid storage treatment but remained undetectable in the compost and pack treatments. However, M. paratuberculosis DNA was detectable through day 56 in all treatments and up to day 175 in liquid storage treatments. Taken together, the results indicate that high-temperature composting is more effective than pack storage or liquid storage of manure in reducing these pathogens in dairy manure. Therefore, thermophilic composting is recommended for treatment of manures destined for pathogen-sensitive environments such as those for vegetable production, residential gardening, or application to rapidly draining fields. PMID:16391093

Engineering of a Stable Whole-Cell Biocatalyst Capable of (S)-Styrene Oxide Formation for Continuous Two-Liquid-Phase Applications

PubMed Central

Panke, Sven; de Lorenzo, Víctor; Kaiser, Arnë; Witholt, Bernard; Wubbolts, Marcel G.

1999-01-01

Recombinant strains of Pseudomonas putida KT2440 carrying genetic expression cassettes with xylene oxygenase- and styrene monooxygenase-encoding genes on their chromosomes could be induced in shaking-flask experiments to specific activities that rivaled those of multicopy-plasmid-based Escherichia coli recombinants. Such strains maintained the introduced styrene oxidation activity in continuous two-liquid-phase cultures for at least 100 generations, although at a lower level than in the shaking-flask experiments. The data suggest that placement of target genes on the chromosome might be a suitable route for the construction of segregationally stable and highly active whole-cell biocatalysts. PMID:10584030

Atmospheric Carbon Dioxide Mixing Ratios from the NOAA CMDL Carbon Cycle Cooperative Global Air Sampling Network (2009)

DOE Data Explorer

Conway, Thomas [NOAA Climate Monitoring and Diagnostics Laboratory, Boulder, CO (USA); Tans, Pieter [NOAA Climate Monitoring and Diagnostics Laboratory, Boulder, CO (USA)

2009-01-01

The National Oceanic and Atmospheric Administration's Climate Monitoring and Diagnostics Laboratory (NOAA/CMDL) has measured CO2 in air samples collected weekly at a global network of sites since the late 1960s. Atmospheric CO2 mixing ratios reported in these files were measured by a nondispersive infrared absorption technique in air samples collected in glass flasks. All CMDL flask samples are measured relative to standards traceable to the World Meteorological Organization (WMO) CO2 mole fraction scale. These measurements constitute the most geographically extensive, carefully calibrated, internally consistent atmospheric CO2 data set available and are essential for studies aimed at better understanding the global carbon cycle budget.

Enhanced Biosynthesis of Withanolides by Elicitation and Precursor Feeding in Cell Suspension Culture of Withania somnifera (L.) Dunal in Shake-Flask Culture and Bioreactor

PubMed Central

Sivanandhan, Ganeshan; Selvaraj, Natesan; Ganapathi, Andy; Manickavasagam, Markandan

2014-01-01

The present study investigated the biosynthesis of major and minor withanolides of Withania somnifera in cell suspension culture using shake-flask culture and bioreactor by exploiting elicitation and precursor feeding strategies. Elicitors like cadmium chloride, aluminium chloride and chitosan, precursors such as cholesterol, mevalonic acid and squalene were examined. Maximum total withanolides detected [withanolide A (7606.75 mg), withanolide B (4826.05 mg), withaferin A (3732.81 mg), withanone (6538.65 mg), 12 deoxy withanstramonolide (3176.63 mg), withanoside IV (2623.21 mg) and withanoside V (2861.18 mg)] were achieved in the combined treatment of chitosan (100 mg/l) and squalene (6 mM) along with 1 mg/l picloram, 0.5 mg/l KN, 200 mg/l L-glutamine and 5% sucrose in culture at 4 h and 48 h exposure times respectively on 28th day of culture in bioreactor. We obtained higher concentrations of total withanolides in shake-flask culture (2.13-fold) as well as bioreactor (1.66-fold) when compared to control treatments. This optimized protocol can be utilized for commercial level production of withanolides from suspension culture using industrial bioreactors in a short culture period. PMID:25089711

Culture of prostate epithelial cells of the rhesus monkey on extracellular matrix substrate: influence of steroids and insulin-like growth factors.

PubMed

Udayakumar, T S; Jeyaraj, D A; Rajalakshmi, M; Sharma, R S

1999-09-01

Rhesus monkey prostate epithelial cells from the cranial lobe were isolated and cultured in flasks coated either with collagen IV or laminin. The effects of stromal cell medium, androgens and growth factors on cell number, thymidine incorporation and secretory activity were assessed. The results indicate that dihydrotestosterone (DHT) and androstenedione have stimulatory influences on cell proliferation and secretion in coated flasks. DHT was more effective in increasing cell number but the induction of secretory activity was similar with both steroids. The combination of IGF-I and -II resulted in inducing better cell proliferation and secretory activity than the individual IGFs but, of the two IGFs, IGF-I was more effective than IGF-II. DHT with IGFs was more potent in inducing proliferation, differentiation and secretion than androstenedione. Even in the absence of steroids or growth factors, colony formation and confluence occurred in coated flasks but cell differentiation and secretion only to a limited extent. In conclusion, we were able to establish an in vitro primary culture of prostate epithelial cells from rhesus monkey using extracellular matrix proteins, steroids and growth factors as additional supplements. This culture system may be useful to study prostate cell physiology and to identify drugs that can inhibit cell proliferation.

Production of Laccase by Recombinant Yarrowia lipolytica from Molasses: Bioprocess Development Using Statistical Modeling and Increase Productivity in Shake-Flask and Bioreactor Cultures.

PubMed

Darvishi, Farshad; Moradi, Marzieh; Madzak, Catherine; Jolivalt, Claude

2017-03-01

Laccases are used in numerous applications, from green degradation of various xenobiotic compounds, waste detoxification, textile dye bleaching, and delignification of lignocellulose materials to biofuel production. In this study, the recombinant Yarrowia lipolytica YL4 strain carrying the white-rot fungus Trametes versicolor laccase IIIb gene was used for laccase production from beet molasses as an agro-industrial residue. Response surface methodology was used to statistical optimization of the production of laccase by Y. lipolytica using an industrial medium containing molasses which allows a six times increase in laccase activity compared to primary medium contains glucose after 144 h. In bioreactor cultivation after 48 h, laccase production reached to 3.7- and 22.5-fold more than optimized and primary media in shake-flask cultures, respectively. Laccase productivity in bioreactor (0.0937 U/h) was higher than shake-flask culture (0.0084 U/h). The present study provides valuable information about statistical optimization of bioprocess development for cost-effective production of laccase and other heterologous proteins in Y. lipolytica from beet molasses as sole carbon source, thus allowing the valorization and decreasing environmental pollution of this agro-industrial waste.

Microarray platform affords improved product analysis in mammalian cell growth studies

PubMed Central

Li, Lingyun; Migliore, Nicole; Schaefer, Eugene; Sharfstein, Susan T.; Dordick, Jonathan S.; Linhardt, Robert J.

2014-01-01

High throughput (HT) platforms serve as cost-efficient and rapid screening method for evaluating the effect of cell culture conditions and screening of chemicals. The aim of the current study was to develop a high-throughput cell-based microarray platform to assess the effect of culture conditions on Chinese hamster ovary (CHO) cells. Specifically, growth, transgene expression and metabolism of a GS/MSX CHO cell line, which produces a therapeutic monoclonal antibody, was examined using microarray system in conjunction with conventional shake flask platform in a non-proprietary medium. The microarray system consists of 60 nl spots of cells encapsulated in alginate and separated in groups via an 8-well chamber system attached to the chip. Results show the non-proprietary medium developed allows cell growth, production and normal glycosylation of recombinant antibody and metabolism of the recombinant CHO cells in both the microarray and shake flask platforms. In addition, 10.3 mM glutamate addition to the defined base media results in lactate metabolism shift in the recombinant GS/MSX CHO cells in the shake flask platform. Ultimately, the results demonstrate that the high-throughput microarray platform has the potential to be utilized for evaluating the impact of media additives on cellular processes, such as, cell growth, metabolism and productivity. PMID:24227746

Inverse modeling of CO2 sources and sinks using satellite observations of CO2 from TES and surface flask measurements

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

Nassar, Ray; Jones, DBA; Kulawik, SS

2011-01-01

We infer CO2 surface fluxes using satellite observations of mid-tropospheric CO2 from the Tropospheric Emission Spectrometer (TES) and measurements of CO2 from surface flasks in a time-independent inversion analysis based on the GEOS-Chem model. Using TES CO2 observations over oceans, spanning 40 S 40 N, we find that the horizontal and vertical coverage of the TES and flask data are complementary. This complementarity is demonstrated by combining the datasets in a joint inversion, which provides better constraints than from either dataset alone, when a posteriori CO2 distributions are evaluated against independent ship and aircraft CO2 data. In particular, the jointmore » inversion offers improved constraints in the tropics where surface measurements are sparse, such as the tropical forests of South America. Aggregating the annual surface-to-atmosphere fluxes from the joint inversion for the year 2006 yields 1.13 0.21 PgC for the global ocean, 2.77 0.20 PgC for the global land biosphere and 3.90 0.29 PgC for the total global natural flux (defined as the sum of all biospheric, oceanic, and biomass burning contributions but excluding CO2 emissions from fossil fuel combustion). These global ocean and global land fluxes are shown to be near the median of the broad range of values from other inversion results for 2006. To achieve these results, a bias in TES CO2 in the Southern Hemisphere was assessed and corrected using aircraft flask data, and we demonstrate that our results have low sensitivity to variations in the bias correction approach. Overall, this analysis suggests that future carbon data assimilation systems can benefit by integrating in situ and satellite observations of CO2 and that the vertical information provided by satellite observations of mid-tropospheric CO2 combined with measurements of surface CO2, provides an important additional constraint for flux inversions.« less

Dynamic collection and analysis of volatile organic compounds from the headspace of cell cultures.

PubMed

Baranska, A; Smolinska, A; Boots, A W; Dallinga, J W; van Schooten, F J

2015-10-15

Exhaled breath has proven to be a valuable source of information about human bodies. Subtle differences between volatile organic compounds (VOCs) formed endogenously can be detected and become a base for a potential monitoring tool for health and disease. Until now, there has been a lack of biological and mechanistic knowledge of the processes involved in the production of relevant VOCs. Among the possible sources of health-related and disease-related VOCs are microorganisms found in the respiratory tract and in the gut. Other VOCs in the body are produced by cells that are influenced by the disease, for instance, due to metabolic disorders and/or inflammation. To gain insight into the in vivo production of VOCs by human cells and thus the exhaled breath composition, in vitro experiments involving relevant cells should be studied because they may provide valuable information on the production of VOCs by the affected cells. To this aim we developed and validated a system for dynamically (continuously) collecting headspace air in vitro using a Caco-2 cell line. The system allows the application of different cell lines as well as different experimental setups, including varying exposure times and treatment options while preserving cell viability. Significant correlation (p  ⩽  0.0001) between collection outputs within each studied group confirmed high reproducibility of the collection system. An example of such an application is presented here. We studied the influence of oxidative stress on the VOC composition of the headspace air of Caco-2 cells. By comparing the VOC composition of air flushed through empty culture flasks (n  =  35), flasks with culture medium (n  =  35), flasks with medium and cells (n  =  20), flasks with medium and an oxidative stressor (H2O2) (n  =  20), and flasks with medium, stressor, and cells (n  =  20), we were able to separate the effects from the stressor on the cells from all other interactions. Measurements were performed with gas chromatography time-of-flight mass spectrometry. Multivariate data analysis allowed detection of significant altered compounds in the compared groups. We found a significant change (p  ⩽  0.001) of the composition of VOCs due to the stressing of the Caco-2 cells by H2O2. A total of ten VOCs showed either increased or decreased abundance in the headspace of the cell cultures due to the presence of the H2O2 stressor.

Secondary metabolism in simulated microgravity

NASA Technical Reports Server (NTRS)

Demain, A. L.; Fang, A.

2001-01-01

We have studied microbial secondary metabolism in a simulated microgravity (SMG) environment provided by NASA rotating-wall bioreactors (RWBs). These reactors were designed to simulate some aspects of actual microgravity that occur in space. Growth and product formation were observed in SMG in all cases studied, i.e., Bacillus brevis produced gramicidin S (GS), Streptomyces clavuligerus made beta-lactam antibiotics, Streptomyces hygroscopicus produced rapamycin, and Escherichia coli produced microcin B17 (MccB17). Of these processes, only GS production was unaffected by SMG; production of the other three products was inhibited. This was determined by comparison with performance in an RWB positioned in a different mode to provide a normal gravity (NG) environment. Carbon source repression by glycerol of the GS process, as observed in shaken flasks, was not observed in the RWBs, whether operated in the SMG or NG mode. The same phenomenon occurred in the case of MccB17 production, with respect to glucose repression. Thus, the negative effects of carbon source on GS and beta-lactam formation are presumably dependent on shear, turbulence, and/or vessel geometry, but not on gravity. Stimulatory effects of phosphate and the precursor L-lysine on beta-lactam antibiotic production, as observed in flasks, also occurred in SMG. An almost complete shift in the localization of produced MccB17 from cells to extracellular medium was observed when E. coli was grown in the RWB under SMG or NG. If a plastic bead was placed in the RWB, accumulation became cellular, as it is in shaken flasks, indicating that sheer stress favors a cellular location. In the case of rapamycin, the same type of shift was observed, but it was less dramatic, i.e., growth in the RWB under SMG shifted the distribution of produced rapamycin from 2/3 cellular:1/3 extracellular to 1/3 cellular:2/3 extracellular. Stress has been shown to induce or promote secondary metabolism in a number of other microbial systems. RWBs provide a low stress SMG environment, which, however, supports only poor production of MccB17, as compared to production in shaken flasks. We wondered whether the poor production in RWBs under SMG is due to the low level of stress, and whether increasing stress in the RWBs would raise the amount of MccB17 formed. We found that increasing shear stress by adding a single Teflon bead to the RWB improved MccB17 production. Although shear stress seems to have a marked positive effect on MccB17 production in SMG, addition of various concentrations of ethanol to RWBs (or to shaken flasks) failed to increase MccB17 production. Ethanol stress merely decreased production and, at higher concentrations, inhibited growth. Interestingly, cells growing in the RWB were much more resistant to the growth- and production-inhibitory effects of ethanol than cells growing in shaken flasks. With respect to S. hygroscopicus, addition of Teflon beads to the RWB reversed the inhibition of growth, but rapamycin production was still markedly inhibited, and the distribution did not revert back to a preferential cellular site.

Effect of stover fraction and storage method on glucose production during enzymatic hydrolysis.

PubMed

Montross, M D; Crofcheck, C L

2004-05-01

One avenue for overcoming the economic challenges associated with the production of ethanol from renewable resources is to reduce the cost of the biomass feedstock. The balance between storage costs and benefits depend on the storage method and composition changes of individual stover fractions. Corn stover from bales stored inside and outside of a barn was separated into an interior and exterior layer after approximately 10 months of storage. The cobs, stalks, and leaves and husks were separated, dried, and ground through a 2 mm screen. Stover, sodium acetate (buffer), cellulase, and deionized water were added to 125 ml flasks. The mixture was held at 50 degrees C in an incubator and samples taken for glucose determination. The average glucose concentration after 60 h of hydrolysis from cobs, leaves and husks, and stalks was 10.5, 9.6, and 3.1 g/l, respectively. Cobs, leaves, and husks produced over 300% more glucose than stalks. Storage outside of the barn decreased the glucose production from individual stover components between 4% and 8%. The effect of stover fraction type on glucose production was significant, while the storage treatment effect was not significant. Fractionation of corn stover may be a method to increase the value of corn stover as a feedstock for glucose production.

Deglycosylation of isoflavones in isoflavone-rich soy germ flour by Aspergillus oryzae KACC 40247.

PubMed

Lee, Seon-Hwa; Seo, Min-Ho; Oh, Deok-Kun

2013-12-11

Aspergillus oryzae KACC 40247 was selected as an efficient daidzein-producing fungus from strains of the genus Aspergillus by using 5% (w/v) soy germ flour (SGF) as an isoflavone-glycoside-rich medium. The culture conditions, including SGF concentration, agitation speed, initial pH, temperature, and time, were optimized as follows: 7% (w/v) SGF, initial pH 6.0, 33 °C, 300 rpm, and 24 h in a 100 mL baffled flask. The determined amount of isoflavone aglycons in SGF using 50% ethyl acetate was the highest among the solvent systems tested and it was 3.7-fold higher than that using 70% methanol. Under the optimized conditions, the content and concentration of daidzein were 134 mg/g of SGF and 9.4 g/L, respectively, with a productivity of 391 ± 2.8 mg/L/h, and those of isoflavone aglycons were 165 mg/g of SGF and 11.5 g/L, respectively, with a productivity of 479 mg/L/h. Optimization of culture conditions increased the content, concentration, and productivity of isoflavone aglycons by 3.1-, 3.0-, and 3.7-fold, respectively. To our knowledge, this is the highest production of isoflavone aglycons reported to date.

Structural and functional characterization of recombinant napin-like protein of Momordica charantia expressed in methylotrophic yeast Pichia pastoris.

PubMed

Yadav, Shailesh Kumar R; Sahu, Tejram; Dixit, Aparna

2016-08-01

Napin and napin-like proteins belong to the 2S albumin seed storage family of proteins and have been shown to display a variety of biological activities. However, due to a high degree of polymorphism, purification of a single napin or napin-like protein exhibiting biological activity is extremely difficult. In the present study, we have produced the napin-like protein of Momordica charantia using the methylotrophic Pichia pastoris expression system. The recombinant napin-like protein (rMcnapin) secreted in the extracellular culture supernatant was enriched by ammonium sulfate precipitation, and purified using size exclusion chromatography at a yield of ∼290 mg/L of culture. Secondary structure analysis of the purified rMcnapin revealed it to be predominantly α-helical with minimal β strand content. CD spectroscopic and fluorescence spectroscopic analyses revealed the rMcnapin to be stable at a wide range of temperatures and pH. The rMcnapin exhibited antifungal activity against Trichoderma viride with an IC50 of ∼3.7 μg/ml and trypsin inhibitor activity with an IC50 of 4.2 μM. Thus, large amounts of homogenous preparations of the biologically active rMcnapin could be obtained at shake flask level, which is otherwise difficult from its natural source.

Rapid and qualitative fluorescence-based method for the assessment of PHA production in marine bacteria during batch culture.

PubMed

Elain, Anne; Le Fellic, Magali; Corre, Yves-Marie; Le Grand, Adélaïde; Le Tilly, Véronique; Audic, Jean-Luc; Bruzaud, Stéphane

2015-10-01

The expansion of polyhydroxyalkanoates (PHAs) into the biodegradable polymers market is mainly prevented by their production process which is still complicated with a low efficiency, resulting in relatively expensive products. In this study, we developed a method that used the lipophilic fluorescent probe Nile Red (1 mg l(-1) solution in DMSO) directly into the culture broth to stain the PHA inclusions inside bacterial cells followed by detection of the emitted fluorescence by both microscopic and spectrometric techniques. Epifluorescence microscopy provides a rapid tool to distinguish producing from non-producing bacterial species and the relative fluorescence intensity (FI) determined at the maximum of emission spectra in the wavelength region of 560-710 nm (λ(ex): 543 nm), allows a fast assessment of the cultural conditions that may enhance PHA production yield. During two-step cultivation in 500-ml flasks with glucose as the sole carbon source, the method aimed to select bacterial strains efficient for PHA synthesis among a marine collection. Subsequently, the NR assay was used to determine the C0/N0 ratio of the producing media that may improve the polymer yield as well as to follow the time course of fermentation. Characterization by GC-MS and DSC confirmed the production of the P(3-HB) homopolymer.

Soluble Prokaryotic Expression and Purification of Bioactive Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand.

PubMed

Do, Bich Hang; Nguyen, Minh Tan; Song, Jung-A; Park, Sangsu; Yoo, Jiwon; Jang, Jaepyeong; Lee, Sunju; So, Seoungjun; Yoon, Yejin; Kim, Inki; Lee, Kyungjin; Jang, Yeon Jin; Choe, Han

2017-12-28

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is considered as an antitumor agent owing to its ability to induce apoptosis of cancer cells without imparting toxicity toward most normal cells. TRAIL is produced in poor yield because of its insoluble expression in the cytoplasm of E. coli . In this study, we achieved soluble expression of TRAIL by fusing maltose-binding protein (MBP), b'a' domain of protein disulfide isomerase (PDIb'a'), or protein disulfide isomerase at the N-terminus of TRAIL. The TRAIL was purified using subsequent immobilized metal affinity chromatography and amylose-binding chromatography, with the tag removal using tobacco etch virus protease. Approximately 4.5 mg of pure TRAIL was produced from 125 ml flask culture with a purification yield of 71.6%. The endotoxin level of the final product was 0.4 EU/μg, as measured by the Limulus amebocyte lysate endotoxin assay. The purified TRAIL was validated and shown to cause apoptosis of HeLa cells with an EC₅₀ and Hill coefficient of 0.6 ± 0.03 nM and 2.41 ± 0.15, respectively. The high level of apoptosis in HeLa cells following administration of purified TRAIL indicates the significance and novelty of this method for producing high-grade and high-yield TRAIL.

Determination of mercury in agroindustrial samples by flow-injection cold vapor atomic absorption spectrometry using ion exchange and reductive elution.

PubMed

Gomes Neto, J A; Zara, L F; Rocha, J C; Santos, A; Dakuzaku, C S; Nóbrega, J A

2000-03-06

A flow-injection system with a Chelite-S(R) cationic resin packed minicolumn is proposed for the determination of trace levels of mercury in agroindustrial samples by cold vapor atomic absorption spectrometry. Improved sensitivity and selectivity are attained since mercuric ions are on-line concentrated whereas other potential interferents are discarded. With on-line reductive elution procedure, concentrated hydrochloric acid could be replaced by 10% w/v SnCl(2), in 6 M HCl as eluent. The reversed-intermittent stream either carries the atomic mercury to the flow cell in the forward direction or removes the residue from reactor/gas-liquid separator to a discarding flask in the opposite direction. Concentration and volume of reagent, acidity, flow rates, commutation times and potential interfering species were investigated. For 120 s preconcentration time, the proposed system handles about 25 samples h(-1) (50.0-500 ng l(-1)), consuming about 10 ml sample and 5 mg SnCl(2) per determination. The detection limit is 0.8 ng l(-1) and the relative standard deviation (RSD) (n=12) of a 76.7 ng l(-1) sample is about 5%. Results are in agreement with certified value of standard materials at 95% confidence level and good recoveries (97-128%) of spiked samples were found.

Raman gas sensing of modified Ag nanoparticle SERS

NASA Astrophysics Data System (ADS)

Myoung, NoSoung; Yoo, Hyung Keun; Hwang, In-Wook

2014-03-01

Recent progress in modified Surface Enhanced Raman Scattering (SERS) using Ag nanoparticles makes them promising optical technique for direct gas sensing of interest. However, SERS has been shown to provide sub ppb level detection of the compounds in the vapor phase. The major problem with the sensitivity scaling-up was in the development of fabrication technology for stability and reproducibility of SERS substrates. We report an optimization of 1-propanethiol coated multiple Ag nanoparticle layers on SiO2 substrate as well as new records of real-time, simultaneous vapor phase detection of toluene and 1-2 dichlorobenzene by the radiation of fiber optic coupled 785 nm diode laser and spectrograph. Multiple depositions of Ag NPs were loaded on SiO2 and soaked in 1-propanethiol solution for 24 hours to modify the surface into hydrophobic due to the characteristics of vapor phase of our interests. Raman bands at 1003 cm-1 and 1130 cm-1 for toluene and 12DCB, respectively were compared to 1089 cm-1 and each gas concentration in 1000 mL flask were calculated as a function of each vapor phase ratio. The saturation of toluene and 12DCB were limited only by 800 ppm and the detectable range was 0.6-800 ppm.

Evaluation of cysteine ethyl ester as efficient inducer for glutathione overproduction in Saccharomyces spp.

PubMed

Lorenz, Eric; Schmacht, Maximilian; Senz, Martin

2016-11-01

Economical yeast based glutathione (GSH) production is a process that is influenced by several factors like raw material and production costs, biomass production and efficient biotransformation of adequate precursors into the final product GSH. Nowadays the usage of cysteine for the microbial conversion into GSH is industrial state of practice. In the following study, the potential of different inducers to increase the GSH content was evaluated by means of design of experiments methodology. Investigations were executed in three natural Saccharomyces strains, S. cerevisiae, S. bayanus and S. boulardii, in a well suited 50ml shake tube system. Results of shake tube experiments were confirmed in traditional baffled shake flasks and finally via batch cultivation in lab-scale bioreactors under controlled conditions. Comprehensive studies showed that the usage of cysteine ethyl ester (CEE) for the batch-wise biotransformation into GSH led up to a more than 2.2 times higher yield compared to cysteine as inducer. Additionally, the intracellular GSH content could be significantly increased for all strains in terms of 2.29±0.29% for cysteine to 3.65±0.23% for CEE, respectively, in bioreactors. Thus, the usage of CEE provides a highly attractive inducing strategy for the GSH overproduction. Copyright © 2016 Elsevier Inc. All rights reserved.

Integrated ozone and biotreatment of pulp mill effluent and changes in biodegradability and molecular weight distribution of organic compounds.

PubMed

Bijan, Leila; Mohseni, Madjid

2005-10-01

The overall effectiveness of integrating ozonation with biological treatment on the biodegradability enhancement and recalcitrant organic matter (ROM) removal from pulp mill alkaline bleach plant effluent was investigated. Ozonation was performed in a semi-batch bubble column reactor at pH of 11 and 4.5. Batch biological treatment was conducted in shake flasks. Samples obtained during the treatments were monitored for BOD5, COD, TOC, and molecular weight distribution. At an ozone dosage of 0.7-0.8 mg O3/mL wastewater, integrated treatment showed about 30% higher TOC mineralization compared to individual ozonation or biotreatment. Ozone treatment enhanced the biodegradability of the effluent (monitored as 21% COD reduction and 13% BOD5 enhancement), allowing for a higher removal of pollutants. The conversion of high molecular weight (HMW) to low molecular weight (LMW) compounds was an important factor in the overall biodegradability enhancement of the alkaline effluent. The overall biodegradability of the LMW compounds did not change over the course of ozonation, but it increased from 5% to 50% (measured as COD removal) for the HMW portion. Ozonation at pH of 11 was more effective than that at pH of 4.5 in terms of generating more biodegradable compounds.

A preliminary study on performance of Saccharomyces cerevisiae n0 DY 7221 immobilized using grafted bioflocculant in bioethanol production

NASA Astrophysics Data System (ADS)

Suci, Windhu Griyasti; Margono, Kaavessina, Mujtahid

2018-02-01

Bioethanol has been well acknowledged to be developed as a biofuel and can be derived from renewable resources. Currently, the utilization of bioethanol as a fuel is more expensive than that of gasoline due to the high production cost. Researchers from industrial and academia have been doing some efforts to reduce it, namely: energy efficiency, exploring many potential renewable resources, increasing fermentation productivity, etc. We propose a novel immobilized Saccharomyces cerevisiae trapped in grafted bioflocculant. The flocculant was developed from polyacrylamide chains grafted into modified starches. This research aims to preliminary performance study of S. cerevisiae immobilized using our new developed method. The bioflocculant solution with the various concentration of 1%, 2%, and 2.5% v/v was dropped into 90 ml of developed inoculum to get flocs which would be used as a starter in fermentation process. The fermentation process was carried out in a shaken flask at 30oC and 150 rpm for 72 hours. The best result was obtained in the sample of 2.5% bioflocculant fraction, i.e. bioethanol 9.25% and enhanced productivity 3.6 times of free cell. These results indicate that flocculation method is a way of immobilizing yeast that needs to be further investigated.

Effect of cell passage on the susceptibility of BALB/3T3 clone A31-1-1 cells to 3-methylcholanthrene-induced morphological transformation

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

Sheu, C.W.; Moreland, F.M.; Dunkel, V.C.

1987-01-01

The response of BALB/3T3 clone A31-1-1 cells to chemically induced morphological transformation was evaluated using 3-methylcholanthrene (MCA). Stock cultures were initiated from cryopreserved cells, grown in T25 flasks containing 5 ml of medium, and replated at subconfluency. Serially transferred cells were then subjected to transformation assay. After 24-hr seeding, cells were incubated 48 hr with MCA in a 5% CO/sub 2/ incubator. They were then rinsed and incubated for an additional 4 weeks with twice weekly medium change. Type III foci were scored after fixation and staining with Giemsa. With serial passage from the frozen state, cells of passages 3-14more » had a low level of spontaneous transformation; zero to 6 type III foci per 20 dishes were counted. In the MCA-treated cultures the number of transformed foci, however, increased with passage. Such passage-related sensitivity to MCA was demonstrated for cells cultured in two batches of sera: one from MA Bioproducts (Lot no. 2E052) and the other from Armour Pharmaceuticals (Lot no. Y65801). The passage-related increase in number of transformed foci was not related to doubling time, cloning efficiency, or MCA-induced growth inhibition.« less

Inventory Control.

ERIC Educational Resources Information Center

Byrum, David L., Ed.

1984-01-01

Describes an electronic thermometer using a precision temperature sensor (includes detailed schematic of circuits) and inexpensive ring holders for round-bottomed flasks. Also describes a method for reducing funnel breakage. (JN)

Process development for the production of 15β-hydroxycyproterone acetate using Bacillus megaterium expressing CYP106A2 as whole-cell biocatalyst.

PubMed

Kiss, Flora M; Lundemo, Marie T; Zapp, Josef; Woodley, John M; Bernhardt, Rita

2015-03-05

CYP106A2 from Bacillus megaterium ATCC 13368 was first identified as a regio- and stereoselective 15β-hydroxylase of 3-oxo-∆4-steroids. Recently, it was shown that besides 3-oxo-∆4-steroids, 3-hydroxy-∆5-steroids as well as di- and triterpenes can also serve as substrates for this biocatalyst. It is highly selective towards the 15β position, but the 6β, 7α/β, 9α, 11α and 15α positions have also been described as targets for hydroxylation. Based on the broad substrate spectrum and hydroxylating capacity, it is an excellent candidate for the production of human drug metabolites and drug precursors. In this work, we demonstrate the conversion of a synthetic testosterone derivative, cyproterone acetate, by CYP106A2 under in vitro and in vivo conditions. Using a Bacillus megaterium whole-cell system overexpressing CYP106A2, sufficient amounts of product for structure elucidation by nuclear magnetic resonance spectroscopy were obtained. The product was characterized as 15β-hydroxycyproterone acetate, the main human metabolite. Since the product is of pharmaceutical interest, our aim was to intensify the process by increasing the substrate concentration and to scale-up the reaction from shake flasks to bioreactors to demonstrate an efficient, yet green and cost-effective production. Using a bench-top bioreactor and the recombinant Bacillus megaterium system, both a fermentation and a transformation process were successfully implemented. To improve the yield and product titers for future industrial application, the main bottlenecks of the reaction were addressed. Using 2-hydroxypropyl-β-cyclodextrin, an effective bioconversion of 98% was achieved using 1 mM substrate concentration, corresponding to a product formation of 0.43 g/L, at a 400 mL scale. Here we describe the successful scale-up of cyproterone acetate conversion from shake flasks to bioreactors, using the CYP106A2 enzyme in a whole-cell system. The substrate was converted to its main human metabolite, 15β-hydroxycyproterone acetate, a highly interesting drug candidate, due to its retained antiandrogen activity but significantly lower progestogen properties than the mother compound. Optimization of the process led to an improvement from 55% to 98% overall conversion, with a product formation of 0.43 g/L, approaching industrial process requirements and a future large-scale application.

Non-invasive and non-destructive measurements of confluence in cultured adherent cell lines.

PubMed

Busschots, Steven; O'Toole, Sharon; O'Leary, John J; Stordal, Britta

2015-01-01

Many protocols used for measuring the growth of adherent monolayer cells in vitro are invasive, destructive and do not allow for the continued, undisturbed growth of cells within flasks. Protocols often use indirect methods for measuring proliferation. Microscopy techniques can analyse cell proliferation in a non-invasive or non-destructive manner but often use expensive equipment and software algorithms. In this method images of cells within flasks are captured by photographing under a standard inverted phase contract light microscope using a digital camera with a camera lens adaptor. Images are analysed for confluence using ImageJ freeware resulting in a measure of confluence known as an Area Fraction (AF) output. An example of the AF method in use on OVCAR8 and UPN251 cell lines is included. •Measurements of confluence from growing adherent cell lines in cell culture flasks is obtained in a non-invasive, non-destructive, label-free manner.•The technique is quick, affordable and eliminates sample manipulation.•The technique provides an objective, consistent measure of when cells reach confluence and is highly correlated to manual counting with a haemocytometer. The average correlation co-efficient from a Spearman correlation (n = 3) was 0.99 ± 0.008 for OVCAR8 (p = 0.01) and 0.99 ± 0.01 for UPN251 (p = 0.01) cell lines.

Scale-up laccase production from Trametes versicolor stimulated by vanillic acid.

PubMed

Wang, Ke-Feng; Hu, Jian-Hua; Guo, Chen; Liu, Chun-Zhao

2016-07-01

An efficient strategy for laccase production in Trametes versicolor cultures was developed using vanillic acid as the inducer. The optimized vanillic acid treatment strategy consisted of exposing 2-day-old mycelia cultures to 80 mg/L vanillic acid. After 4 days, laccase activity of 588.84 U/L was achieved in flasks which represented a 1.79-fold increase compared to the control. In 200-L airlift bioreactor, the maximal laccase activity reached up to 785.12 U/L using the optimized vanillic acid treatment strategy. The zymograms of culture supernatants revealed three bands with laccase activity, among which Lac1 and Lac2 were abundant laccase isoforms constitutively expressed, and Lac3 was an inducible isozyme by vanillic acid. The results of real-time quantitative PCR showed that the transcription level of lcc in T. versicolor cultures grown with vanillic acid for 7 days was about 5.64-fold greater than that without vanillic acid in flasks. In 200-L airlift bioreactor cultures of T. versicolor with addition of vanillic acid, the transcript level of lcc at day 7 was 2.62-fold higher than that in flasks with vanillic acid due to the good mass transfer and oxygen supply in the bioreactor system. This study provides a basis for understanding the induction mechanism of vanillic acid for laccase production and has good potential for industrial applications.

Study of plant phototropic responses to different LEDs illumination in microgravity

NASA Astrophysics Data System (ADS)

Zyablova, Natalya; Berkovich, Yuliy A.; Skripnikov, Alexander; Nikitin, Vladimir

2012-07-01

The purpose of the experiment planned for Russian BION-M #1, 2012, biosatellite is research of Physcomitrella patens (Hedw.) B.S.G. phototropic responses to different light stimuli in microgravity. The moss was chosen as small-size higher plant. The experimental design involves five lightproof culture flasks with moss gametophores fixed inside the cylindrical container (diameter 120 mm; height 240 mm). The plants in each flask are illuminated laterally by one of the following LEDs: white, blue (475 nm), red (625 nm), far red (730 nm), infrared (950 nm). The gametophores growth and bending are captured periodically by means of five analogue video cameras and recorder. The programmable command module controls power supply of each camera and each light source, commutation of the cameras and functioning of video recorder. Every 20 minutes the recorder is sequentially connecting to one of the cameras. This results in a clip, containing 5 sets of frames in a row. After landing time-lapse films are automatically created. As a result we will have five time-lapse films covering transformations in each of the five culture flasks. Onground experiments demonstrated that white light induced stronger gametophores phototropic bending as compared to red and blue stimuli. The comparison of time-lapse recordings in the experiments will provide useful information to optimize lighting assemblies for space plant growth facilities.

An innovative iterative thresholding algorithm for tumour segmentation and volumetric quantification on SPECT images: Monte Carlo-based methodology and validation.

PubMed

Pacilio, M; Basile, C; Shcherbinin, S; Caselli, F; Ventroni, G; Aragno, D; Mango, L; Santini, E

2011-06-01

Positron emission tomography (PET) and single-photon emission computed tomography (SPECT) imaging play an important role in the segmentation of functioning parts of organs or tumours, but an accurate and reproducible delineation is still a challenging task. In this work, an innovative iterative thresholding method for tumour segmentation has been proposed and implemented for a SPECT system. This method, which is based on experimental threshold-volume calibrations, implements also the recovery coefficients (RC) of the imaging system, so it has been called recovering iterative thresholding method (RIThM). The possibility to employ Monte Carlo (MC) simulations for system calibration was also investigated. The RIThM is an iterative algorithm coded using MATLAB: after an initial rough estimate of the volume of interest, the following calculations are repeated: (i) the corresponding source-to-background ratio (SBR) is measured and corrected by means of the RC curve; (ii) the threshold corresponding to the amended SBR value and the volume estimate is then found using threshold-volume data; (iii) new volume estimate is obtained by image thresholding. The process goes on until convergence. The RIThM was implemented for an Infinia Hawkeye 4 (GE Healthcare) SPECT/CT system, using a Jaszczak phantom and several test objects. Two MC codes were tested to simulate the calibration images: SIMIND and SimSet. For validation, test images consisting of hot spheres and some anatomical structures of the Zubal head phantom were simulated with SIMIND code. Additional test objects (flasks and vials) were also imaged experimentally. Finally, the RIThM was applied to evaluate three cases of brain metastases and two cases of high grade gliomas. Comparing experimental thresholds and those obtained by MC simulations, a maximum difference of about 4% was found, within the errors (+/- 2% and +/- 5%, for volumes > or = 5 ml or < 5 ml, respectively). Also for the RC data, the comparison showed differences (up to 8%) within the assigned error (+/- 6%). ANOVA test demonstrated that the calibration results (in terms of thresholds or RCs at various volumes) obtained by MC simulations were indistinguishable from those obtained experimentally. The accuracy in volume determination for the simulated hot spheres was between -9% and 15% in the range 4-270 ml, whereas for volumes less than 4 ml (in the range 1-3 ml) the difference increased abruptly reaching values greater than 100%. For the Zubal head phantom, errors ranged between 9% and 18%. For the experimental test images, the accuracy level was within +/- 10%, for volumes in the range 20-110 ml. The preliminary test of application on patients evidenced the suitability of the method in a clinical setting. The MC-guided delineation of tumor volume may reduce the acquisition time required for the experimental calibration. Analysis of images of several simulated and experimental test objects, Zubal head phantom and clinical cases demonstrated the robustness, suitability, accuracy, and speed of the proposed method. Nevertheless, studies concerning tumors of irregular shape and/or nonuniform distribution of the background activity are still in progress.

Fluid mechanics of spinner-flask bioreactors

NASA Astrophysics Data System (ADS)

Sucosky, Philippe; Neitzel, G. Paul

2000-11-01

The dynamic environment within bioreactors used for in vitro tissue growth has been observed to affect the development of mammalian cells. Many studies have shown that moderate mechanical stress enhances growth of some tissues whereas high shear levels and turbulence seem to damage cells. In order to optimize the design and the operating conditions of bioreactors, it is important to understand the fluid-dynamic characteristics and to control the stress levels within these devices. The present research focuses on the characterization of the flow field within a spinner-flask bioreactor. The dynamic properties of the flow are investigated experimentally using particle-image velocimetry with a refractive-index-matched model. Phase-locked ensemble-averaging is employed to provide some information on the turbulence characteristics of the model culture medium in the vicinity of a model tissue construct.

Mercury recycling in the United States in 2000

USGS Publications Warehouse

Brooks, William E.; Matos, Grecia R.

2005-01-01

Reclamation and recycling of mercury from used mercury- containing products and treatment of byproduct mercury from gold mining is vital to the continued, though declining, use of this metal. Mercury is reclaimed from mercury-containing waste by treatment in multistep high-temperature retorts-the mercury is volatized and then condensed for purification and sale. Some mercury-containing waste, however, may be landfilled, and landfilled material represents loss of a recyclable resource and a threat to the environment. Related issues include mercury disposal and waste management, toxicity and human health, and regulation of mercury releases in the environment. End-users of mercury-containing products may face fines and prosecution if these products are improperly recycled or not recycled. Local and State environmental regulations require adherence to the Resource Conservation and Recovery Act and the Comprehensive Environmental Response, Compensation, and Liability Act to regulate generation, treatment, and disposal of mercury-containing products. In the United States, several large companies and a number of smaller companies collect these products from a variety of sources and then reclaim and recycle the mercury. Because mercury has not been mined as a principal product in the United States since 1992, mercury reclamation from fabricated products has become the main source of mercury. Principal product mercury and byproduct mercury from mining operations are considered to be primary materials. Mercury may also be obtained as a byproduct from domestic or foreign gold-processing operations. In the early 1990s, U.S. manufacturers used an annual average that ranged from 500 to 600 metric tons of recycled and imported mercury for fabrication of automobile convenience switches, dental amalgam, fluorescent lamps, medical uses and thermometers, and thermostats. The amount now used for fabrication is estimated to be 200 metric tons per year or less. Much of the data on mercury is estimated because it is a low-volume commodity and its production, use, and disposal is difficult to track. The prices and volumes of each category of mercury-containing material may change dramatically from year to year. For example, the average price of mercury was approximately $150 per flask from 2000 until 2003 and then rose sharply to $650 per flask in fall 2004 and approximately $850 per flask in spring 2005. Since 1927, the common unit for measuring and pricing mercury has been the flask in order to conform to the system used at Almaden, Spain (Meyers, 1951). One flask weighs 34.5 kilograms, and 29 flasks of mercury are contained in a metric ton. In the United States, the chlorine-caustic soda industry, which is the leading end-user of elemental mercury, recycles most of its mercury in-plant as home scrap. Annual purchases of replacement mercury by the chlorine-caustic soda industry indicate that some mercury may be lost through evaporation to the environment, put into a landfill as industrial waste, or trapped within pipes in the plant. Impending closure of domestic and foreign mercury-cell chlorine-caustic soda plants and the shift to nonmercury technology for chlorine-caustic soda production could ultimately result in a significant volume of elemental mercury for recycling, sale, or storage. Globally, mercury is widely used in artisanal, or small-scale, gold mining. Most of that mercury is lost to the environment and is not recycled. The recycling rate for mercury was not available owing to insufficient data in 2000, and the efficiency of mercury recycling was estimated to be 62 percent.

Antifoam addition to shake flask cultures of recombinant Pichia pastoris increases yield

PubMed Central

2011-01-01

Background Pichia pastoris is a widely-used host for recombinant protein production. Initial screening for both suitable clones and optimum culture conditions is typically carried out in multi-well plates. This is followed by up-scaling either to shake-flasks or continuously stirred tank bioreactors. A particular problem in these formats is foaming, which is commonly prevented by the addition of chemical antifoaming agents. Intriguingly, antifoams are often added without prior consideration of their effect on the yeast cells, the protein product or the influence on downstream processes such as protein purification. In this study we characterised, for the first time, the effects of five commonly-used antifoaming agents on the total amount of recombinant green fluorescent protein (GFP) secreted from shake-flask cultures of this industrially-relevant yeast. Results Addition of defined concentrations of Antifoam A (Sigma), Antifoam C (Sigma), J673A (Struktol), P2000 (Fluka) or SB2121 (Struktol) to shake-flask cultures of P. pastoris increased the total amount of recombinant GFP in the culture medium (the total yield) and in the case of P2000, SB2121 and J673A almost doubled it. When normalized to the culture density, the GFP specific yield (μg OD595-1) was only increased for Antifoam A, Antifoam C and J673A. Whilst none of the antifoams affected the growth rate of the cells, addition of P2000 or SB2121 was found to increase culture density. There was no correlation between total yield, specific yield or specific growth rate and the volumetric oxygen mass transfer coefficient (kLa) in the presence of antifoam. Moreover, the antifoams did not affect the dissolved oxygen concentration of the cultures. A comparison of the amount of GFP retained in the cell by flow cytometry with that in the culture medium by fluorimetry suggested that addition of Antifoam A, Antifoam C or J673A increased the specific yield of GFP by increasing the proportion secreted into the medium. Conclusions We show that addition of a range of antifoaming agents to shake flask cultures of P. pastoris increases the total yield of the recombinant protein being produced. This is not only a simple method to increase the amount of protein in the culture, but our study also provides insight into how antifoams interact with microbial cell factories. Two mechanisms are apparent: one group of antifoams (Antifoam A, Antifoam C and J673A) increases the specific yield of GFP by increasing the total amount of protein produced and secreted per cell, whilst the second (P2000 or SB2121) increases the total yield by increasing the density of the culture. PMID:21426555

How-To-Do-It: Plant Regeneration.

ERIC Educational Resources Information Center

Pietraface, William J.

1988-01-01

Describes a procedure for the growth of tobacco plants in flasks. Demonstrates plant tissue culture manipulation, totipotency, and plant regeneration in approximately 12 weeks. Discusses methods, materials, and expected results. (CW)

Cytotoxic effects of some animal and vegetable extracts and some chemicals on liver and colon carcinoma and myosarcoma.

PubMed

Bayazit, Vahdettin

2004-02-01

To study, the cytotoxic effects of some biological and chemical agents on G1, S, G2, M and G0 phases of liver and colon carcinomas and myosarcoma cells obtained with chemical carcinogens dimethylbenzanthracene (DMBA) and cadmium chloride. Eight rabbit livers, colon carcinoma and myosarcoma cell lines were obtained by injection of DMBA in the Biology Laboratory, of the University of Dumlupinar, Kutahya, Turkey between January 2001 and June 2003. All lines were grown at 37 degrees celsius and 5% carbon dioxide in sterile RPMI-1640 medium with 10% fetal bovine serum after addition of glutamate, penicillin (50 units/ml) and streptomycin (50 ug/ml) (complete medium). Cells were grown on standard tissue culture plastic flasks to 80% confluence and passed by trypsinization. Tortoise (Testudo graeca) shell, sponge (Geodia cydonium), medusa (Aurelia aurita), meat flies (Calliphora erythrocephala) larva, frog (Rana ridibunda) larva and juniper (Juniperus communis) berry extracts killed a large amount of the liver and colon carcinomas and the myosarcoma cells in G2, M and G0 phases (p<0.01). The mistletoe (Viscum album) extract had more effect in only the G0 phase (p<0.05). Genistein, genistin, glycitein, glycitin, daitzein and daitzin have significantly decreased in the cancer cells tests, particularly, genistein and daitzein caused the apoptotic effect in G2, M and G0 phases (p<0.01). Cesium chloride, a mixture of cesium chloride with magnesium chloride had the most effect on tumor cells (p<0.01). AzhexSi, Azhex-AzhepSi, Et-Azhex-AzhepSi, AzhepSi, Hexamine and DL 54 have been inhibited in various levels of the cancer cells (p<0.05, p<0.01). This data suggest that some biological extracts and chemicals tested may be useful chemotherapeutic agents to inhibit the growth of cancer cells. This study sheds some light for new anti cancerogenic experiments preventing various cancers on humans.

Nutritional demands and metabolic characteristics of the DSIR-HA-1179 insect cell line during growth and infection with the Oryctes nudivirus.

PubMed

Pushparajan, Charlotte; Claus, Juan Daniel; Marshall, Sean D G; Visnovsky, Gabriel

2017-12-01

The DSIR-HA-1179 coleopteran cell line has been identified as a susceptible and permissive host for the in vitro replication of the Oryctes nudivirus, which can be used as a biopesticide against the coconut rhinoceros beetle, pest of palms. The major challenge to in vitro large-scale Oryctes nudivirus production is ensuring process economy. This rests, among other requisites, on the use of low-cost culture media tailored to the nutritional and metabolic needs of the cell line, both in uninfected and infected cultures. The aim of the present study was to characterize the nutritional demands and the metabolic characteristics of the DSIR-HA-1179 cell line during growth and subsequent infection with Oryctes nudivirus in the TC-100 culture medium. Serum-supplementation of the culture medium was found to be critical for cell growth, and addition of 10% fetal bovine serum v/v led to a maximum viable cell density (16.8 × 10 5 cells ml -1 ) with a population doubling time of 4.2 d. Nutritional and metabolic characterization of the cell line revealed a trend of glucose and glutamine consumption but minimal uptake of other amino acids, negligible production of lactate and ammonia, and the accumulation of alanine, both before and after infection. The monitoring of virus production kinetics showed that the TC-100 culture medium was nutritionally sufficient to give a peak yield of 7.38 × 10 7 TCID 50 ml -1 of OrNV at the 6th day post-infection in attached cultures of DSIR-HA-1179 cells in 25 cm 2 T-flasks. Knowledge of the cell line's nutritional demands and virus production kinetics will aid in the formulation of a low-cost culture medium and better process design for large-scale OrNV production in future.

Spontaneously Generating Life in Your Classroom? Pasteur, Spallanzani and Science Process.

ERIC Educational Resources Information Center

Byington, Scott

2001-01-01

Presents an experiment that tests for spontaneous generation, or abiogenesis. Observes microbial growth in nutrient broth under seven different flask environments. Includes instructions for the methods. (YDS)

Cross-section perimeter is a suitable parameter to describe the effects of different baffle geometries in shaken microtiter plates

PubMed Central

2014-01-01

Background Biotechnological screening processes are performed since more than 8 decades in small scale shaken bioreactors like shake flasks or microtiter plates. One of the major issues of such reactors is the sufficient oxygen supply of suspended microorganisms. Oxygen transfer into the bulk liquid can in general be increased by introducing suitable baffles at the reactor wall. However, a comprehensive and systematic characterization of baffled shaken bioreactors has never been carried out so far. Baffles often differ in number, size and shape. The exact geometry of baffles in glass lab ware like shake flasks is very difficult to reproduce from piece to piece due to the hard to control flow behavior of molten glass during manufacturing. Thus, reproducibility of the maximum oxygen transfer capacity in such baffled shake flasks is hardly given. Results As a first step to systematically elucidate the general effect of different baffle geometries on shaken bioreactor performance, the maximum oxygen transfer capacity (OTRmax) in baffled 48-well microtiter plates as shaken model reactor was characterized. This type of bioreactor made of plastic material was chosen, as the exact geometry of the baffles can be fabricated by highly reproducible laser cutting. As a result, thirty different geometries were investigated regarding their maximum oxygen transfer capacity (OTRmax) and liquid distribution during shaking. The relative perimeter of the cross-section area as new fundamental geometric key parameter is introduced. An empirical correlation for the OTRmax as function of the relative perimeter, shaking frequency and filling volume is derived. For the first time, this correlation allows a systematic description of the maximum oxygen transfer capacity in baffled microtiter plates. Conclusions Calculated and experimentally determined OTRmax values agree within ± 30% accuracy. Furthermore, undesired out-of-phase operating conditions can be identified by using the relative perimeter as key parameter. Finally, an optimum well geometry characterized by an increased perimeter of 10% compared to the unbaffled round geometry is identified. This study may also assist to comprehensively describe and optimize the baffles of shake flasks in future. PMID:25093039

Effects of Water Vapor on the Data Quality of the Stable Oxygen Isotopic Ratio of Atmospheric Carbon Dioxide

NASA Astrophysics Data System (ADS)

Evans, C. U.; White, J. W.; Vaughn, B.; Tans, P. P.; Pardo, L.

2007-12-01

The stable oxygen isotopic ratio of carbon dioxide can potentially track fundamental indicators of environmental change such as the balance between photosynthesis and respiration on regional to global scales. The Stable Isotope Laboratory (SIL) at the Institute of Arctic and Alpine Research (INSTAAR), University of Colorado at Boulder, has measured the stable isotopes of atmospheric carbon dioxide from more than 60 NOAA/Earth System Research Laboratory (ESRL) air flask-sampling sites since the early 1990s. If air is sampled without drying, oxygen can exchange between carbon dioxide and water in the flasks, entirely masking the desired signal. An attempt to investigate how water vapor is affecting the δ18O signal is accomplished by comparing the SIL measurements with specific humidity, calculated from the National Climatic Data Center (NCDC) global integrated surface hourly temperature and dew point database, at the time of sampling. Analysis of sites where samples have been collected initially without drying, and subsequently with a drying kit, in conjunction with the humidity data, has led to several conclusions. Samples that initially appear isotopically unaltered, in that their δ18O values are within the expected range, are being subtly influenced by the water vapor in the air. At Bermuda and other tropical to semi-tropical sites, the 'wet' sampling values have a seasonal cycle that is strongly anti-correlated to the specific humidity, while the 'dry' values have a seasonal cycle that is shifted earlier than the specific humidity cycle by 1-2 months. The latter phasing is expected given the seasonal phasing between climate over the ocean and land, while the former is consistent with a small, but measurable isotope exchange in the flasks. In addition, we note that there is a strong (r > 0.96) correlation between the average specific humidity and the percent of rejected samples for 'wet' sampling. This presents an opportunity for determining a threshold of specific humidity, below which air flask samples can be trusted. This approach may allow segregation of suspect and trusted data, and thus provide an improved record of oxygen isotopic ratios of carbon dioxide over the past two decades.

A hollow definitive obturator fabrication technique for management of partial maxillectomy.

PubMed

Patil, Pravinkumar Gajanan; Patil, Smita Pravinkumar

2012-11-01

Maxillary obturator prosthesis is the most frequent treatment option for management of partial or total maxillectomy. Heavy weight of the obturators is often a dislocating factor. Hollowing the prosthesis to reduce its weight is the well established fact. The alternate technique to hollow-out the prosthesis has been described in this article which is a variation of previously described processing techniques. A pre-shaped wax-bolus was incorporated inside the flasks during packing of the heat-polymerized acrylic resin to automatically create the hollow space. The processing technique described is a single step flasking procedure to construct a closed-hollow-obturator prosthesis as a single unit. To best understand the technique, this article describes management of a patient who had undergone partial maxillectomy secondary to squamous cell carcinoma rehabilitated with a hollow-obturator prosthesis.

A hollow definitive obturator fabrication technique for management of partial maxillectomy

PubMed Central

Patil, Smita Pravinkumar

2012-01-01

Maxillary obturator prosthesis is the most frequent treatment option for management of partial or total maxillectomy. Heavy weight of the obturators is often a dislocating factor. Hollowing the prosthesis to reduce its weight is the well established fact. The alternate technique to hollow-out the prosthesis has been described in this article which is a variation of previously described processing techniques. A pre-shaped wax-bolus was incorporated inside the flasks during packing of the heat-polymerized acrylic resin to automatically create the hollow space. The processing technique described is a single step flasking procedure to construct a closed-hollow-obturator prosthesis as a single unit. To best understand the technique, this article describes management of a patient who had undergone partial maxillectomy secondary to squamous cell carcinoma rehabilitated with a hollow-obturator prosthesis. PMID:23236579

Morphological changes in human melanoma cells following irradiation with thermal neutrons.

PubMed

Barkla, D H; Allen, B J; Brown, J K; Mountford, M; Mishima, Y; Ichihashi, M

1989-01-01

Morphological changes in two human melanoma cell lines, MM96 and MM418, following irradiation with thermal neutrons, were studied using light and electron microscopy. The results show that the response of human malignant melanoma cells to neutron irradiation is both cell line dependent and dose dependent, and that in any given cell line, some cells are more resistant to irradiation than others, thus demonstrating heterogeneity in respect to radiosensitivity. Cells repopulating MM96 flasks after irradiation were morphologically similar to the cells of origin whereas in MM418 flasks cells differentiated into five morphologically distinct subgroups and showed increased melanization. The results also show that radiation causes distinctive morphological patterns of damage although ultrastructural changes unique to the high LET particles released from boron 10 neutron capture are yet to be identified.

[Analysis of false-positive reaction for bacterial detection of blood samples with the automated microbial detection system BacT/ALERT 3D].

PubMed

Zhu, Li-Wei; Yang, Xue-Mei; Xu, Xiao-Qin; Xu, Jian; Lu, Huang-Jun; Yan, Li-Xing

2008-10-01

This study was aimed to analyze the results of false positive reaction in bacterial detection of blood samples with BacT/ALERT 3D system, to evaluate the specificity of this system, and to decrease the false positive reaction. Each reaction flasks in past five years were processed for bacteria isolation and identification. When the initial cultures were positive, the remaining samples and the corresponding units were recultured if still available. 11395 blood samples were detected. It is worthy of note that the incubator temperature should be stabilized, avoiding fluctuation; when the cultures were alarmed, the reaction flasks showed be kept some hours for further incubation so as to trace a sharply increasing signal to support the judgement of true bacterial growth. The results indicated that 122 samples (1.07%) wee positive at initial culture, out of them 107 samples (88.7%) were found bacterial, and 15 samples (12.3%) were found nothing. The detection curves of positive samples resulted from bacterial growth showed ascent. In conclusion, maintenance of temperature stability and avoidance of temperature fluctuation in incubator could decrease the occurrence of false-positive reaction in detection process. The reaction flasks with positive results at initial culture should be recultured, and whether existence of a sharply ascending logarilhimic growth phase in bacterial growth curve should be further detected, which are helpful to distinguish false-positive reactions from true positive, and thus increase the specificity of the BacT/ALERT system.

Bench-scale evaluation of in situ bioremediation strategies for soil at a former manufactured gas plant site.

PubMed

Li, Jun; Pignatello, Joseph J; Smets, Barth F; Grasso, Domenico; Monserrate, Esteban

2005-03-01

We examined the biodegradation and desorption of a set of 15 polycyclic aromatic hydrocarbon (PAH) compounds in coal tar-contaminated soil at a former manufactured gas plant site to evaluate the feasibility of in situ bioremediation. Experiments were conducted in well-mixed aerobic soil suspensions containing various additives over a 93- to 106-d period. In general, both biotransformation and desorption decreased with PAH ring size, becoming negligible for the six-ring PAH compounds. Biodegradation by indigenous microorganisms was strongly accelerated by addition of inorganic nutrients (N, P, K, and trace metals). The rates of biotransformation of PAH compounds by indigenous microorganisms in nutrient-amended flasks outpaced their maximum (i.e., chelate-enhanced) rates of desorption to an infinite sink (Tenax) in sterilized systems run in parallel, suggesting that indigenous organisms facilitated desorption. Biodegradation by indigenous organisms in nutrient-amended flasks appeared to be unaffected by the addition of a site-derived bacterial enrichment culture, resulting in approximately 100-fold higher aromatic dioxygenase levels, and by the addition of 0.01 M chelating agent (citrate or pyrophosphate), although such chelating agents greatly enhanced desorption in microbially inactivated flasks. The strong ability of nutrients to enhance degradation of the bioavailable PAHs indicates that their persistence for many decades at this site likely results from nutrient-limited natural biodegradation, and it also suggests that an effective strategy for their bioremediation could consist simply of adding inorganic nutrients.

Optimization of Ex Vivo Murine Bone Marrow Derived Immature Dendritic Cells: A Comparative Analysis of Flask Culture Method and Mouse CD11c Positive Selection Kit Method

PubMed Central

Salwe, Sukeshani; Kothari, Sweta; Chowdhary, Abhay; Deshmukh, Ranjana A.

2018-01-01

12–14 days of culturing of bone marrow (BM) cells containing various growth factors is widely used method for generating dendritic cells (DCs) from suspended cell population. Here we compared flask culture method and commercially available CD11c Positive Selection kit method. Immature BMDCs' purity of adherent as well as suspended cell population was generated in the decreasing concentration of recombinant-murine granulocyte-macrophage colony-stimulating factor (rmGM-CSF) in nontreated tissue culture flasks. The expression of CD11c, MHCII, CD40, and CD86 was measured by flow cytometry. We found significant difference (P < 0.05) between the two methods in the adherent cells population but no significant difference was observed between the suspended cell populations with respect to CD11c+ count. However, CD11c+ was significantly higher in both adhered and suspended cell population by culture method but kit method gave more CD11c+ from suspended cells population only. On the other hand, using both methods, immature DC expressed moderate level of MHC class II molecules as well as low levels of CD40 and CD86. Our findings suggest that widely used culture method gives the best results in terms of yield, viability, and purity of BMDCs from both adherent and suspended cell population whereas kit method works well for suspended cell population. PMID:29682352

First 2 years of Atmospheric CO2 measurements in the Estany Llong plain (2100 masl, Parc Nacional d'Aigüestortes i Estany de Sant Maurici, Pyrenees, Catalonia, Spain).

NASA Astrophysics Data System (ADS)

Curcoll, Roger; Recolons, Montserrat; Font, Anna; Agraz, Laura; Parga, Elena; Bacardit, Montse; Camarero, Lluís.; Pueyo, Salva; Rodó, Xavier; Morguí, Josep Anton

2010-05-01

Since April 2009, air samples are being taken bi-weekly at 10 GMT in the plain of the Estany Llong at 2100 masl. Estany Llong air sampling site (ELL, 42°34'29''N 0°57'17''E) is a remote site situated in the SW principal valley of the Parc Nacional d'Aigüestortes i Estany de Sant Maurici. New Flask-sampling equipment for Remote Mountain Sites was developed by the Institut Català de Ciències del Clima (IC3) to allow flask sampling in extreme weather conditions and carrying the sampling equipment for more than 10 km without damaging flasks. Dry Air analysis for CO2 are done at the Laboratory of IC3 using two coupled modified IRGA Licor-7000, where both pressure and flow are externally controlled. Far away from populated areas, ELL site acts as a remote site, but it is also responding to discrete events as snow melting, summer cattle breeding on pastures and trekking frequentation. Series of CO2 obtained are included as part of Long Term Ecological Research (LTER) at the Parc Nacional d'Aigüestortes i Estany de Sant Maurici. In the long term, these measurements show the mountain ecosystems contribution and geomorphologic influence on the CO2 budget of the air masses crossing a mountain range.

Atmospheric Methane Mixing Ratios--The NOAA/CMDL Global Cooperative Air Sampling Network\\, 1983-1993

DOE Data Explorer

Dlugokencky, E. J. [National Oceanic and Atmospheric Administration, Boulder, Colorado (USA); Lang, P. M. [National Oceanic and Atmospheric Administration, Boulder, Colorado (USA); Masarie, K. A. [National Oceanic and Atmospheric Administration, Boulder, Colorado (USA); Steele, L. P. [Commonwealth Scientific and Industrial Research Organisation, Aspendale, Victoria, Australia

1994-01-01

This data base presents atmospheric methane (CH4) mixing ratios from flask air samples collected over the period 1983-1993 by the National Oceanic and Atmospheric Administration, Climate Monitoring and Diagnostics Laboratory's (NOAA/CMDL's) global cooperative air sampling network. Air samples were collected approximately once per week at 44 fixed sites (37 of which were still active at the end of 1993). Samples were also collected at 5 degree latitude intervals along shipboard cruise tracks in the Pacific Ocean between North America and New Zealand (or Australia) and at 3 degree latitude intervals along cruise tracks in the South China Sea between Singapore and Hong Kong. The shipboard measurements were made approximately every 3 weeks per latitude zone by each of two ships in the Pacific Ocean and approximately once every week per latitude zone in the South China Sea. All samples were analyzed for CH4 at the NOAA/CMDL laboratory in Boulder, Colorado, by gas chromatography with flame ionization detection, and each aliquot was referenced to the NOAA/CMDL methane standard scale. In addition to providing the complete set of atmospheric CH4 measurements from flask air samples collected at the NOAA/CMDL network sites, this data base also includes files which list monthly mean mixing ratios derived from the individual flask air measurements. These monthly summary data are available for 35 of the fixed sites and 21 of the shipboard sampling sites.

Simultaneous Extraction Optimization and Analysis of Flavonoids from the Flowers of Tabernaemontana heyneana by High Performance Liquid Chromatography Coupled to Diode Array Detector and Electron Spray Ionization/Mass Spectrometry

PubMed Central

Sathishkumar, Thiyagarajan; Baskar, Ramakrishnan; Aravind, Mohan; Tilak, Suryanarayanan; Deepthi, Sri; Bharathikumar, Vellalore Maruthachalam

2013-01-01

Flavonoids are exploited as antioxidants, antimicrobial, antithrombogenic, antiviral, and antihypercholesterolemic agents. Normally, conventional extraction techniques like soxhlet or shake flask methods provide low yield of flavonoids with structural loss, and thereby, these techniques may be considered as inefficient. In this regard, an attempt was made to optimize the flavonoid extraction using orthogonal design of experiment and subsequent structural elucidation by high-performance liquid chromatography-diode array detector-electron spray ionization/mass spectrometry (HPLC-DAD-ESI/MS) techniques. The shake flask method of flavonoid extraction was observed to provide a yield of 1.2 ± 0.13 (mg/g tissue). With the two different solvents, namely, ethanol and ethyl acetate, tried for the extraction optimization of flavonoid, ethanol (80.1 mg/g tissue) has been proved better than ethyl acetate (20.5 mg/g tissue). The optimal conditions of the extraction of flavonoid were found to be 85°C, 3 hours with a material ratio of 1 : 20, 75% ethanol, and 1 cycle of extraction. About seven different phenolics like robinin, quercetin, rutin, sinapoyl-hexoside, dicaffeic acid, and two unknown compounds were identified for the first time in the flowers of T. heyneana. The study has also concluded that L16 orthogonal design of experiment is an effective method for the extraction of flavonoid than the shake flask method. PMID:25969771

Optimization of Ex Vivo Murine Bone Marrow Derived Immature Dendritic Cells: A Comparative Analysis of Flask Culture Method and Mouse CD11c Positive Selection Kit Method.

PubMed

Gosavi, Rahul Ashok; Salwe, Sukeshani; Mukherjee, Sandeepan; Dahake, Ritwik; Kothari, Sweta; Patel, Vainav; Chowdhary, Abhay; Deshmukh, Ranjana A

2018-01-01

12-14 days of culturing of bone marrow (BM) cells containing various growth factors is widely used method for generating dendritic cells (DCs) from suspended cell population. Here we compared flask culture method and commercially available CD11c Positive Selection kit method. Immature BMDCs' purity of adherent as well as suspended cell population was generated in the decreasing concentration of recombinant-murine granulocyte-macrophage colony-stimulating factor (rmGM-CSF) in nontreated tissue culture flasks. The expression of CD11c, MHCII, CD40, and CD86 was measured by flow cytometry. We found significant difference ( P < 0.05) between the two methods in the adherent cells population but no significant difference was observed between the suspended cell populations with respect to CD11c+ count. However, CD11c+ was significantly higher in both adhered and suspended cell population by culture method but kit method gave more CD11c+ from suspended cells population only. On the other hand, using both methods, immature DC expressed moderate level of MHC class II molecules as well as low levels of CD40 and CD86. Our findings suggest that widely used culture method gives the best results in terms of yield, viability, and purity of BMDCs from both adherent and suspended cell population whereas kit method works well for suspended cell population.

Fabrication of viable and functional pre-vascularized modular bone tissues by coculturing MSCs and HUVECs on microcarriers in spinner flasks.

PubMed

Zhang, Songjie; Zhou, Min; Ye, Zhaoyang; Zhou, Yan; Tan, Wen-Song

2017-08-01

Slow vascularization often impedes the viability and function of engineered bone replacements. Prevascularization is a promising way to solve this problem. In this study, a new process was developed by integrating microcarrier culture and coculture to fabricate pre-vascularized bone microtissues with mesenchymal stem cells (MSCs) and human umbilical vein endothelial cells (HUVECs). Initially, coculture medium and cell ratio between MSCs and HUVECs were optimized in tissue culture plates concerning cell proliferation, osteogenesis and angiogenesis. Subsequently, cells were seeded onto CultiSpher S microcarriers in spinner flasks and subjected to a two-stage (proliferative-osteogenic) culture process for four weeks. Both cells proliferated and functioned well in chosen medium and a 1 : 1 ratio between MSCs and HUVECs was chosen for better angiogenesis. After four weeks of culture in spinner flasks, the microtissues were formed with high cellularity, evenly distributed cells and tube formation ability. While coculture with HUVECs exerted an inhibitory effect on osteogenic differentiation of MSCs, with downregulated alkaline phosphatase activity, mineralization and gene expression of COLI, RUNX2 and OCN, this could be attenuated by employing a delayed seeding strategy of HUVECs against MSCs during the microtissue fabrication process. Collectively, this work established an effective method to fabricate pre-vascularized bone microtissues, which would lay a solid foundation for subsequent development of vascularized tissue grafts for bone regeneration. Copyright © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Metals in Metal Salts: A Copper Mirror Demonstration

ERIC Educational Resources Information Center

Pike, Robert D.

2010-01-01

A simple lecture demonstration is described to show the latent presence of metal atoms in a metal salt. Copper(II) formate tetrahydrate is heated in a round-bottom flask forming a high-quality copper mirror.

75 FR 67714 - Notice of Intent To Suspend Certain Pesticide Registrations

Federal Register 2010, 2011, 2012, 2013, 2014

2010-11-03

.... August 30, 2008.. No data received. coefficient (n- octanol/water) shake flask method. 830.7570 Partition December 14, 2007. December 24, 2007. August 30, 2008.. No data received. coefficient (n- octanol/water...

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

PubMed

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

2016-01-20

Fructo-oligosaccharides (FOS) obtained by fermentation of sucrose may be purified at large-scale by continuous chromatography (Simulated Moving Bed: SMB). In order to improve the efficiency of the subsequent SMB purification, the optimization of the fermentative broth composition in salts and sugars was investigated. Fermentations conducted at reduced amount of salts, using Aureobasidium pullulans whole cells, yielded 0.63 ± 0.03 g of FOS per gram of initial sucrose. Additionally, a microbial treatment was proposed to reduce the amount of small saccharides in the mixture. Two approaches were evaluated, namely a co-culture of A. pullulans with Saccharomyces cerevisiae; and a two-step fermentation in which FOS were first synthesized by A. pullulans and then the small saccharides were metabolized by S. cerevisiae. Assays were performed in 100mL shaken flasks and further scaled-up to a 3 L working volume bioreactor. Fermentations in two-step were found to be more efficient than the co-culture ones. FOS were obtained with a purity of 81.6 ± 0.8% (w/w), on a dry weight basis, after the second-step fermentation with S. cerevisiae. The sucrose amount was reduced from 13.5 to 5.4% in total sugars, which suggests that FOS from this culture broth will be more efficiently separated by SMB. Copyright © 2015 Elsevier Ltd. All rights reserved.

Suppression of pyritic sulphur during flotation tests using the bacterium Thiobacillus ferrooxidans.

PubMed

Townsley, C C; Atkins, A S; Davis, A J

1987-07-01

Environmental concern about sulphur dioxide emissions has led to the examination of the possibility of removing pyritic sulphur from coal prior to combustion during froth flotation, a routine method for coal cleaning at the pit-head. The bacterium Thiobacillus ferrooxidans was effective in leaching 80% and 63% -53 mum pyrite at 2% and 6% pulp density in shake flasks in 240 and 340 h, respectively.The natural floatability of pyrite was significantly reduced in the Hallimond tube following 2.5 min of conditioning in membrane-filtered bacterial liquor prior to flotation. The suppression effect was greatly enhanced in the presence of Thiobacillus ferrooxidans. A bacterial suspension in pH 2.0 distilled water showed 85% suppression, whereas in spent growth liquor this value was 95%. The optimum bacterial density was 3.25 x 10(10) cells/g pyrite in 230-ml distilled water (2% pulp density) in the Hallimond tube. The degree of suppression by the cells was related to particle size but not to pH or temperature. The sulphur content of a synthetic coal/pyrite mixture was reduced from 10.9 to 2.1% by flotation after bacterial preconditioning. It is postulated that pyrite removal in coals which are cleaned by froth flotation could be significantly reduced using a bacterial preconditioning stage with a short residence time of 2.5 min.

[Using Excess Activated Sludge Treated 4-Chlorophenol Contained Waste Water to Cultivate Chlorella vulgaris].

PubMed

Wang, Lu; Chen, Xiu-rong; Yan, Long; He, Yi-xuan; Shi, Zhen-dong

2015-04-01

Using different rations of sludge extracts and supernate from 4-Chlorophenol (4-CP) simulated wastewater's excess sludge after centrifugation to cultivate the Chlorella vulgaris to achieve the goal of excess sludge utilization together with chlorella cultivating. The experiments were performed in 500 mL flasks with different rations of sludge extracts & BG-11 and supernate & BG-11 in a light growth chamber respectively. Number of algal cells, Chlorophyll, enzyme activity, oil and water total nitrogen (TN), total phosphorus (TP), total organic carbon (TOC), toxicity index were investigated. Result showed that the nutrition supplies and toxicity in the excess sludge were removed efficiently via Chlorella vulgaris, the removal rates of TN and TP were at least 40% and 90% respectively; After 10 days cultivation, the density growth of 50% sludge extracts was 20 times higher of the beginning while its chlorophyll content was lower than that of the blank group. Sludge extracts could promote the proliferation of algae, but were not conducive to the synthesis of chlorophyll. The quantity of SOD in per cell showed Chlorella vulgaris gave a positive response via stimulation from toxicant in sludge extracts and supernate. The best time for collecting chlorella vulgaris was the fifth day of cultivation, taking neutral oil accumulation as the evaluating indicator for its utilization combined with the removal of supplies and toxicity.

Ultrasound assisted dialysis of semi-permeable membrane devices for the simultaneous analysis of a wide number of persistent organic pollutants.

PubMed

Bustamante, Julen; Navarro, Patricia; Arana, Gorka; de Diego, Alberto; Madariaga, Juan Manuel

2013-09-30

A new procedure based on ultrasound assisted dialysis (UAD) for the simultaneous and quantitative extraction of a wide number of persistent organic pollutants (POPs) such as polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) or some other organochlorinated pesticides (OCPs) contained in semi-permeable membrane devices (SPMDs) has been developed. This extraction technique combines the advantages of the organic solvent dialysis (OSD) and the speed of the ultrasound assisted extraction. The extraction was performed in an ultrasound bath for 32 min placing the SPMD in a glass flask covered with 80 mL of hexane. This set-up is able to extract simultaneously up to 8 samples. The proposed method entails good repeatabilities (RSD 2-13%) and recoveries (around 100% for almost every analyte). Limits of detection were at ng SPMD(-1) level and enough for the determination of the target analytes in a slightly polluted aquatic environment, as it was tested by successfully comparing the OSD to the proposed methodology. Therefore, the results obtained show that the UAD can be a good alternative for the extraction of POPs in SPMDs as it requires short extraction times and solvent volumes, and provides a cleaner extract for the subsequent clean-up step. Moreover, it fits better than the OSD to the general requirements of Green Chemistry. Copyright © 2013 Elsevier B.V. All rights reserved.

Construction of a highly thermostable 1,3-1,4-β-glucanase by combinational mutagenesis and its potential application in the brewing industry.

PubMed

Niu, Chengtuo; Zhu, Linjiang; Hill, Annie; Alex Speers, R; Li, Qi

2017-01-01

To improve the thermostability and catalytic property of a mesophilic 1,3-1,4-β-glucanase by combinational mutagenesis and to test its effect in congress mashing. A mutant β-glucanase (rE-BglTO) constructed by combinational mutagenesis showed a 25 °C increase in optimal temperature (to 70 °C) a 19.5 °C rise in T 50 value and a 15.6 °C increase in melting temperature compared to wild-type enzyme. Its half-life values at 60 and 70 °C were 152 and 99 min, which were 370 and 800 % higher than those of wild-type enzyme. Besides, its specific activity and k cat value were 42,734 U mg -1 and 189 s -1 while its stability under acidic conditions was also improved. In flask fermentation, the catalytic activity of rE-BglTO reached 2381 U ml -1 , which was 63 % higher than that of wild-type enzyme. The addition of rE-BglTO in congress mashing decreased the filtration time and viscosity by 21.3 and 9.6 %, respectively. The mutant β-glucanase showed high catalytic activity and thermostability which indicated that rE-BglTO is a good candidate for application in the brewing industry.

In silico and in vitro prediction of gastrointestinal absorption from potential drug eremantholide C.

PubMed

Caldeira, Tamires G; Saúde-Guimarães, Dênia A; Dezani, André B; Serra, Cristina Helena Dos Reis; de Souza, Jacqueline

2017-11-01

Analysis of the biopharmaceutical properties of eremantholide C, sesquiterpene lactone with proven pharmacological activity and low toxicity, is required to evaluate its potential to become a drug. Preliminary analysis of the physicochemical characteristics of eremantholide C was performed in silico. Equilibrium solubility was evaluated using the shake-flask method, at 37.0 °C, 100 rpm during 72 h in biorelevant media. The permeability was analysed using parallel artificial membrane permeability assay, at 37.0 °C, 50 rpm for 5 h. The donor compartment was composed of an eremantholide C solution in intestinal fluid simulated without enzymes, while the acceptor compartment consisted of phosphate buffer. Physicochemical characteristics predicted in silico indicated that eremantholide C has a low solubility and high permeability. In-vitro data of eremantholide C showed low solubility, with values for the dose/solubility ratio (ml): 9448.82, 10 389.61 e 15 000.00 for buffers acetate (pH 4.5), intestinal fluid simulated without enzymes (pH 6.8) and phosphate (pH 7.4), respectively. Also, it showed high permeability, with effective permeability of 30.4 × 10 -6 cm/s, a higher result compared with propranolol hydrochloride (9.23 × 10 -6 cm/s). The high permeability combined with its solubility, pharmacological activity and low toxicity demonstrate the importance of eremantholide C as a potential drug candidate. © 2017 Royal Pharmaceutical Society.

A method for the solvent extraction of low-boiling-point plant volatiles.

PubMed

Xu, Ning; Gruber, Margaret; Westcott, Neil; Soroka, Julie; Parkin, Isobel; Hegedus, Dwayne

2005-01-01

A new method has been developed for the extraction of volatiles from plant materials and tested on seedling tissue and mature leaves of Arabidopsis thaliana, pine needles and commercial mixtures of plant volatiles. Volatiles were extracted with n-pentane and then subjected to quick distillation at a moderate temperature. Under these conditions, compounds such as pigments, waxes and non-volatile compounds remained undistilled, while short-chain volatile compounds were distilled into a receiving flask using a high-efficiency condenser. Removal of the n-pentane and concentration of the volatiles in the receiving flask was carried out using a Vigreux column condenser prior to GC-MS. The method is ideal for the rapid extraction of low-boiling-point volatiles from small amounts of plant material, such as is required when conducting metabolic profiling or defining biological properties of volatile components from large numbers of mutant lines.

Dynamic cell culture on porous biopolymer microcarriers in a spinner flask for bone tissue engineering: a feasibility study.

PubMed

Jin, Guang-Zhen; Park, Jeong-Hui; Seo, Seog-Jin; Kim, Hae-Won

2014-07-01

Porous microspherical carriers have great promise for cell culture and tissue engineering. Dynamic cultures enable more uniform cell population and effective differentiation than static cultures. Here we applied dynamic spinner flask culture for the loading and multiplication of cells onto porous biopolymer microcarriers. The abilities of the microcarriers to populate cells and to induce osteogenic differentiation were examined and the feasibility of in vivo delivery of the constructs was addressed. Over time, the porous microcarriers enabled cell adhesion and expansion under proper dynamic culture conditions. Osteogenic markers were substantially expressed by the dynamic cell cultures. The cell-cultured microcarriers implanted in the mouse subcutaneous tissue for 4 weeks showed excellent tissue compatibility, with minimal inflammatory signs and significant induction of bone tissues. This first report on dynamic culture of porous biopolymer microcarriers providing an effective tool for bone tissue engineering.

Globally Averaged Atmospheric CFC-11 Concentrations: Monthly and Annual Data for the Period 1975-1992 (DB1010)

DOE Data Explorer

Khalil, M. A.K. [Oregon Graduate Institute of Science and Technology Portland, Oregon (USA); Rasmussen, R. A. [Oregon Graduate Institute of Science and Technology Portland, Oregon

1996-01-01

This data set presents globally averaged atmospheric concentrations of chlorofluorocarbon 11, known also as CFC-11 or F-11 (chemical name: trichlorofluoromethane; formula: CCl3F). The monthly global average data are derived from flask air samples collected at eight sites in six locations over the period August 1980-July 1992. The sites are Barrow (Alaska), Cape Meares (Oregon), Cape Kumukahi and Mauna Loa (Hawaii), Cape Matatula (American Samoa), Cape Grim (Tasmania), Palmer Station, and the South Pole (Antarctica). At each collection site, monthly averages were obtained from three flask samples collected every week. In addition to the monthly global averages available for 1980-992, this data set also contains annual global average data for 1975-1985. These annual global averages were derived from January measurements at the South Pole and in the Pacific Northwest of the United States (specifically, Washington state and the Oregon coast).

Effective ethanol production from whey powder through immobilized E. coli expressing Vitreoscilla hemoglobin.

PubMed

Sar, Taner; Stark, Benjamin C; Yesilcimen Akbas, Meltem

2017-03-04

Ethanol production from whey powder was investigated by using free as well as alginate immobilized E. coli and E. coli expressing Vitreoscilla hemoglobin (VHb) in both shake flask and fermenter cultures. Media with varying levels of whey (lactose contents of 3%, 5%, 8% or 15%) and yeast extract (0.3% or 0.5%) were evaluated with fermentation times of 48-96 h. Immobilization and VHb expression resulted in higher ethanol production with all media; the increases ranged from 2% to 89% for immobilization and from 2% to 182% for VHb expression. It was determined that growth medium containing 8% lactose with 0.5% yeast extract yielded the highest ethanol production for free or immobilized strains, with or without VHb expression, in both shake flask and fermenter cultures. Immobilization with alginate was found to be a promising process for ethanol production by VHb-expressing ethanologenic E. coli.

Effective ethanol production from whey powder through immobilized E. coli expressing Vitreoscilla hemoglobin

PubMed Central

Sar, Taner; Stark, Benjamin C.; Yesilcimen Akbas, Meltem

2017-01-01

ABSTRACT Ethanol production from whey powder was investigated by using free as well as alginate immobilized E. coli and E. coli expressing Vitreoscilla hemoglobin (VHb) in both shake flask and fermenter cultures. Media with varying levels of whey (lactose contents of 3%, 5%, 8% or 15%) and yeast extract (0.3% or 0.5%) were evaluated with fermentation times of 48–96 h. Immobilization and VHb expression resulted in higher ethanol production with all media; the increases ranged from 2% to 89% for immobilization and from 2% to 182% for VHb expression. It was determined that growth medium containing 8% lactose with 0.5% yeast extract yielded the highest ethanol production for free or immobilized strains, with or without VHb expression, in both shake flask and fermenter cultures. Immobilization with alginate was found to be a promising process for ethanol production by VHb-expressing ethanologenic E. coli. PMID:27579556

Optimization of growth for the hyperthermophilic archaeon Aeropyrum pernix on a small-batch scale.

PubMed

Milek, Igor; Cigic, Blaz; Skrt, Mihaela; Kaletunç, Gönül; Ulrih, Natasa Poklar

2005-09-01

Growth of Aeropyrum pernix, the first reported aerobic neutrophilic hyperthermophilic archaeon, was investigated under different cultivation parameters. Different sources of seawater, pH, and the cultivation methods were tested with the aim to improve the biomass production. A 1-L glass flask fitted with a condenser and air diffuser was used as a bioreactor. The optimum conditions for maximizing A. pernix biomass were obtained when Na2S2O3.5H2O (1 g/L) with added marine broth 2216 at pH 7.0 (20 mmol HEPES buffer/L) was used as a growing medium in a 1-L flask. The biomass production was 0.45 g dry cell mass/L in 40 h under the optimum conditions, which is more than the 0.42 g dry cell mass/L in 60 h previously obtained.

Atmospheric CO2 Concentrations--The Canadian Background Air Pollution Monitoring Network (1993) (NDP-034)

DOE Data Explorer

Trivett, N. B. A. [Environment Canada, Atmospheric Environment Service, Downsview, Ontario, Canada; Hudec, V. C. [Environment Canada, Atmospheric Environment Service, Downsview, Ontario, Canada; Wong, C. S. [Marine Carbon Research Centre, Institute of Ocean Sciences, Sidney, British Columbia, Canada

1993-01-01

Flask air samples collected at roughly weekly intervals at three Canadian sites [Alert, Northwest Territories (July 1975 through July 1992); Sable Island, Nova Scotia (March 1975 through July 1992); and Cape St. James, British Columbia (May 1979 through July 1992)] were analyzed for CO2 concentration with the measurements directly traceable to the WMO primary CO2 standards. Each record includes the date, atmospheric CO2 concentration, and flask classification code. They provide an accurate record of CO2 concentration levels in Canada during the past two decades. Because these data are directly traceable to WMO standards, this record may be compared with records from other Background Air Pollution Monitoring Network (BAPMoN) stations. The data are in three files (one for each of the monitoring stations) ranging in size from 9.4 to 20.1 kB.

Using box models to quantify zonal distributions and emissions of halocarbons in the background atmosphere.

NASA Astrophysics Data System (ADS)

Elkins, J. W.; Nance, J. D.; Dutton, G. S.; Montzka, S. A.; Hall, B. D.; Miller, B.; Butler, J. H.; Mondeel, D. J.; Siso, C.; Moore, F. L.; Hintsa, E. J.; Wofsy, S. C.; Rigby, M. L.

2015-12-01

The Halocarbons and other Atmospheric Trace Species (HATS) of NOAA's Global Monitoring Division started measurements of the major chlorofluorocarbons and nitrous oxide in 1977 from flask samples collected at five remote sites around the world. Our program has expanded to over 40 compounds at twelve sites, which includes six in situ instruments and twelve flask sites. The Montreal Protocol for Substances that Deplete the Ozone Layer and its subsequent amendments has helped to decrease the concentrations of many of the ozone depleting compounds in the atmosphere. Our goal is to provide zonal emission estimates for these trace gases from multi-box models and their estimated atmospheric lifetimes in this presentation and make the emission values available on our web site. We plan to use our airborne measurements to calibrate the exchange times between the boxes for 5-box and 12-box models using sulfur hexafluoride where emissions are better understood.

Single-Pass, Closed-System Rapid Expansion of Lymphocyte Cultures for Adoptive Cell Therapy

PubMed Central

Klapper, Jacob A.; Thomasian, Armen A.; Smith, Douglas M.; Gorgas, Gayle C.; Wunderlich, John R.; Smith, Franz O.; Hampson, Brian S.; Rosenberg, Steven A.; Dudley, Mark E.

2009-01-01

Adoptive cell therapy (ACT) for metastatic melanoma involves the ex vivo expansion and re-infusion of tumor infiltrating lymphocytes (TIL) obtained from resected specimens. With an overall objective response rate of fifty-six percent, this T-cell immunotherapy provides an appealing alternative to other therapies, including conventional therapies with lower response rates. However, there are significant regulatory and logistical concerns associated with the ex vivo activation and large scale expansion of these cells. The best current practice uses a rapid expansion protocol (REP) consisting of an ex vivo process that occurs in tissue culture flasks (T-flasks) and gas-permeable bags, utilizes OKT3 (anti-CD3 monoclonal antibody), recombinant human interleukin-2, and irradiated peripheral blood mononuclear cells to initiate rapid lymphocyte growth. A major limitation to the widespread delivery of therapy to large numbers of melanoma patients is the open system in which a REP is initiated. To address this problem, we have investigated the initiation, expansion and harvest at clinical scale of TIL in a closed-system continuous perfusion bioreactor. Each cell product met all safety criteria for patient treatment and by head-to-head comparison had a similar potency and phenotype as cells grown in control T-flasks and gas-permeable bags. However, the currently available bioreactor cassettes were limited in the total cell numbers that could be generated. This bioreactor may simplify the process of the rapid expansion of TIL under stringent regulatory conditions thereby enabling other institutions to pursue this form of ACT. PMID:19389403

In vitro cartilage construct generation from silk fibroin- chitosan porous scaffold and umbilical cord blood derived human mesenchymal stem cells in dynamic culture condition.

PubMed

Agrawal, Parinita; Pramanik, Krishna; Biswas, Amit; Ku Patra, Ranjan

2018-02-01

Cartilage construct generation includes a scaffold with appropriate composition to mimic matrix of the damaged tissue on which the stem cells grow and differentiate. In this study, umbilical cord blood (UCB) derived human mesenchymal stem cells (hMSCs) were seeded on freeze dried porous silk-fibroin (SF)/chitosan (CS) scaffolds. Influence of static and dynamic (spinner flask bioreactor) culture conditions on the developing cartilage construct were studied by in-vitro characterization for viability, proliferation, distribution, and chondrogenic differentiation of hMSCs over the scaffold. Constructs developed in spinner flask consisted of 62% live cells, and exhibited 543% more cell density at the core than constructs cultured in static system. Quantification of DNA and glycosaminoglycans accumulation after 21 days showed the progression of chondrogenic differentiation of hMSCs was higher in dynamic culture compared to static one. In constructs generated under dynamic condition, histology staining for proteoglycan matrix, and fluorescence staining for collagen-II and aggrecan showed positive correlation between early and late stage chondrogenic markers, which was further confirmed by quantitative PCR analysis, showing low collagen-I expression and highly expressed Sox9, collagen-II and aggrecan. The present study demonstrated that construct generated by combining 3D SF/CS scaffold with UCB-hMSCs under dynamic condition using spinner flask bioreactor can be used for cartilage tissue regeneration for future medical treatments. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 397-407, 2018. © 2017 Wiley Periodicals, Inc.

Unique differentiation profile of mouse embryonic stem cells in rotary and stirred tank bioreactors.

PubMed

Fridley, Krista M; Fernandez, Irina; Li, Mon-Tzu Alice; Kettlewell, Robert B; Roy, Krishnendu

2010-11-01

Embryonic stem (ES)-cell-derived lineage-specific stem cells, for example, hematopoietic stem cells, could provide a potentially unlimited source for transplantable cells, especially for cell-based therapies. However, reproducible methods must be developed to maximize and scale-up ES cell differentiation to produce clinically relevant numbers of therapeutic cells. Bioreactor-based dynamic culture conditions are amenable to large-scale cell production, but few studies have evaluated how various bioreactor types and culture parameters influence ES cell differentiation, especially hematopoiesis. Our results indicate that cell seeding density and bioreactor speed significantly affect embryoid body formation and subsequent generation of hematopoietic stem and progenitor cells in both stirred tank (spinner flask) and rotary microgravity (Synthecon™) type bioreactors. In general, high percentages of hematopoietic stem and progenitor cells were generated in both bioreactors, especially at high cell densities. In addition, Synthecon bioreactors produced more sca-1(+) progenitors and spinner flasks generated more c-Kit(+) progenitors, demonstrating their unique differentiation profiles. cDNA microarray analysis of genes involved in pluripotency, germ layer formation, and hematopoietic differentiation showed that on day 7 of differentiation, embryoid bodies from both bioreactors consisted of all three germ layers of embryonic development. However, unique gene expression profiles were observed in the two bioreactors; for example, expression of specific hematopoietic genes were significantly more upregulated in the Synthecon cultures than in spinner flasks. We conclude that bioreactor type and culture parameters can be used to control ES cell differentiation, enhance unique progenitor cell populations, and provide means for large-scale production of transplantable therapeutic cells.

Large-scale progenitor cell expansion for multiple donors in a monitored hollow fibre bioreactor.

PubMed

Lambrechts, Toon; Papantoniou, Ioannis; Rice, Brent; Schrooten, Jan; Luyten, Frank P; Aerts, Jean-Marie

2016-09-01

With the increasing scale in stem cell production, a robust and controlled cell expansion process becomes essential for the clinical application of cell-based therapies. The objective of this work was the assessment of a hollow fiber bioreactor (Quantum Cell Expansion System from Terumo BCT) as a cell production unit for the clinical-scale production of human periosteum derived stem cells (hPDCs). We aimed to demonstrate comparability of bioreactor production to standard culture flask production based on a product characterization in line with the International Society of Cell Therapy in vitro benchmarks and supplemented with a compelling quantitative in vivo bone-forming potency assay. Multiple process read-outs were implemented to track process performance and deal with donor-to-donor-related variation in nutrient needs and harvest timing. The data show that the hollow fiber bioreactor is capable of robustly expanding autologous hPDCs on a clinical scale (yield between 316 million and 444 million cells starting from 20 million after ± 8 days of culture) while maintaining their in vitro quality attributes compared with the standard flask-based culture. The in vivo bone-forming assay on average resulted in 10.3 ± 3.7% and 11.0 ± 3.8% newly formed bone for the bioreactor and standard culture flask respectively. The analysis showed that the Quantum system provides a reproducible cell expansion process in terms of yields and culture conditions for multiple donors. Copyright © 2016 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

Trichloroethylene biodegradation by mesophilic and psychrophilic ammonia oxidizers and methanotrophs in groundwater microcosms.

PubMed Central

Moran, B N; Hickey, W J

1997-01-01

This study investigated the efficiency of methane and ammonium for stimulating trichloroethylene (TCE) biodegradation in groundwater microcosms (flasks and batch exchange columns) at a psychrophilic temperature (12 degrees C) typical of shallow aquifers in the northern United States or a mesophilic temperature (24 degrees C) representative of most laboratory experiments. After 140 days, TCE biodegradation rates by ammonia oxidizers and methanotrophs in mesophilic flask microcosms were similar (8 to 10 nmol day-1), but [14C]TCE mineralization (biodegradation to 14CO2) by ammonia oxidizers was significantly greater than that by methanotrophs (63 versus 53%). Under psychrophilic conditions, [14C]TCE mineralization in flask systems by ammonia oxidizers and methanotrophs was reduced to 12 and 5%, respectively. In mesophilic batch exchange columns, average TCE biodegradation rates for methanotrophs (900 nmol liter-1 day-1) were not significantly different from those of ammonia oxidizers (775 nmol liter-1 day-1). Psychrophilic TCE biodegradation rates in the columns were similar with both biostimulants and averaged 145 nmol liter-1 day-1. Methanotroph biostimulation was most adversely affected by low temperatures. At 12 degrees C, the biodegradation efficiencies (TCE degradation normalized to microbial activity) of methanotrophs and ammonia oxidizers decreased by factors of 2.6 and 1.6, respectively, relative to their biodegradation efficiencies at 24 degrees C. Collectively, these experiments demonstrated that in situ bioremediation of TCE is feasible at the psychrophilic temperatures common in surficial aquifers in the northern United States and that for such applications biostimulation of ammonia oxidizers could be more effective than has been previously reported. PMID:9327550

40 CFR Appendix 2 to Subpart A of... - Drilling Fluids Toxicity Test (EPA Method 1619)

Code of Federal Regulations, 2014 CFR

2014-07-01

... any excess air removed by flushing the storage containers with nitrogen under pressure anytime the... on an analytical balance, adding the chemical to a 100-milliliter volumetric flask, and bringing the...

40 CFR Appendix 2 to Subpart A of... - Drilling Fluids Toxicity Test (EPA Method 1619)

Code of Federal Regulations, 2012 CFR

2012-07-01

... any excess air removed by flushing the storage containers with nitrogen under pressure anytime the... on an analytical balance, adding the chemical to a 100-milliliter volumetric flask, and bringing the...

40 CFR Appendix 2 to Subpart A of... - Drilling Fluids Toxicity Test (EPA Method 1619)

Code of Federal Regulations, 2013 CFR

2013-07-01

... any excess air removed by flushing the storage containers with nitrogen under pressure anytime the... on an analytical balance, adding the chemical to a 100-milliliter volumetric flask, and bringing the...

Apparatus for Teaching Physics.

ERIC Educational Resources Information Center

Mamola, Karl C., Ed.

1993-01-01

Discusses how to build an inexpensive Young's modulus apparatus and also a lens with an adjustable focal length. Describes the demonstration of moving a peeled, hard-boiled egg in and out of a flask whose neck is slightly smaller than the egg. (MVL)

40 CFR 799.6755 - TSCA partition coefficient (n-octanol/water), shake flask method.

Code of Federal Regulations, 2012 CFR

2012-07-01

... fact that the P becomes dependent upon the concentration of the solution. Because of the multiple... Potential of Organic Chemicals in Fish. Environmental Science and Technology 8:1113 (1974). (2) Leo, A. et...

13. VIEW, LOOKING WEST FROM LEFT TO RIGHT, SHOWING HIGH ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

13. VIEW, LOOKING WEST FROM LEFT TO RIGHT, SHOWING HIGH PRESSURE AIR FLASK ROOM AND PUMP ROOM - U.S. Naval Submarine Base, New London Submarine Escape Training Tank, Albacore & Darter Roads, Groton, New London County, CT

21 CFR 178.3910 - Surface lubricants used in the manufacture of metallic articles.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 10 minutes. (The reaction between the sulfoxide and the acid is exothermic. Release pressure after...,” above) until siphon action occurs and then refill the tube body. Supply heat to the boiling flask and...

Increasing L-threonine production in Escherichia coli by engineering the glyoxylate shunt and the L-threonine biosynthesis pathway.

PubMed

Zhao, Hui; Fang, Yu; Wang, Xiaoyuan; Zhao, Lei; Wang, Jianli; Li, Ye

2018-04-30

L-threonine is an important amino acid that can be added in food, medicine, or feed. Here, the influence of glyoxylate shunt on an L-threonine producing strain Escherichia coli TWF001 has been studied. The gene iclR was deleted, and the native promoter of the aceBA operon was replaced by the trc promoter in the chromosome of TWF001, the resulting strainTWF004 could produce 0.39 g L-threonine from1 g glucose after 36-h flask cultivation. Further replacing the native promoter of aspC by the trc promoter in the chromosome of TWF004 resulted in the strain TWF006. TWF006 could produce 0.42 g L-threonine from 1 g glucose after 36-h flask cultivation. Three key genes in the biosynthetic pathway of L-threonine, thrA * (a mutated thrA), thrB, and thrC were overexpressed in TWF006, resulting the strain TWF006/pFW01-thrA * BC. TWF006/pFW01-thrA * BC could produce 0.49 g L-threonine from 1 g glucose after 36-h flask cultivation. Next, the genes asd, rhtA, rhtC, or thrE were inserted into the plasmid TWF006/pFW01-thrA * BC, and TWF006 was transformed with these plasmids, resulting the strains TWF006/pFW01-thrA * BC-asd, TWF006/pFW01-thrA * BC-rhtA, TWF006/pFW01-thrA * BC-rhtC, and TWF006/pFW01-thrA * BC-thrE, respectively. These four strains could produce more L-threonine than the control strain, and the highest yield was produced by TWF006/pFW01-thrA * BC-asd; after 36-h flask cultivation, TWF006/pFW01-thrA * BC-asd could produce 15.85 g/l L-threonine, i.e., 0.53 g L-threonine per 1 g glucose, which is a 70% increase relative to the control strain TWF001. The results suggested that the combined engineering of glyoxylate shunt and L-threonine biosynthesis pathway could significantly increase the L-threonine production in E. coli.

Simulation of Local Seismic Ground Motions from the FLASK Underground Nuclear Explosion near the Source Physics Experiment Dry Alluvium Geology Site

NASA Astrophysics Data System (ADS)

Rodgers, A. J.; Pitarka, A.; Wagoner, J. L.; Helmberger, D. V.

2017-12-01

The FLASK underground nuclear explosion (UNE) was conducted in Area 2 of Yucca Flat at the Nevada Test Site on May 26, 1970. The yield was 105 kilotons (DOE/NV-209-Rev 16) and the working point was 529 m below the surface. This test was detonated in faulted Tertiary volcanic rocks of Yucca Flat. Coincidently, the FLASK UNE ground zero (GZ) is close (< 600 m) to the U2ez hole where the Source Physics Experiment will be conducting Phase II of its chemical high explosives test series in the so-called Dry Alluvium Geology (DAG) site. Ground motions from FLASK were recorded by twelve (12) three-component seismic stations in the near-field at ranges 3-4 km. We digitized the paper records and used available metadata on peak particle velocity measurements made at the time to adjust the amplitudes. These waveforms show great variability in amplitudes and waveform complexity with azimuth from the shot, likely due to along propagation path structure such as the geometry of the hard-rock/alluvium contact above the working point. Peak particle velocities at stations in the deeper alluvium to the north, east and south of GZ have larger amplitudes than those to the west where the basement rock is much shallower. Interestingly, the transverse components show a similar trend with azimuth. In fact, the transverse component amplitudes are similar to the other components for many stations overlying deeper basement. In this study, we simulated the seismic response at the available near-field stations using the SW4 three-dimensional (3D) finite difference code. SW4 can simulate seismic wave propagation in 3D inelastic earth structure, including surface topography. SW4 includes vertical mesh refinement which greatly reduces the computational resources needed to run a specific problem. Simulations are performed on high-performance computers with grid spacing as small as 10 meters and resolution to 6 Hz. We are testing various subsurface models to identify the role of 3D structure on path propagation effects from the source. We are also testing 3D models to constrain structure for the upcoming DAG experiments in 2018.

Production of cellulase from Pestalotiopsis versicolor

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

Rao, M.N.A.; Mithal, B.M.; Thakur, R.N.

1983-01-01

Production of cellulase from Pestalotiopsis versicolor was studied in a shake flask culture. The cellulase system was found to be rich in beta-glucosidase. Kinetic parameters such as pH and temperature have been optimized for the various enzyme components. 9 references.

40 CFR Appendix A-5 to Part 60 - Test Methods 11 through 15A

Code of Federal Regulations, 2014 CFR

2014-07-01

... are needed. For the first, third, and fourth impingers, use the Greenburg-Smith design, modified by... of the flask. For the second impinger, use the Greenburg-Smith design with the standard tip. 6.1.3...

40 CFR Appendix A-5 to Part 60 - Test Methods 11 through 15A

Code of Federal Regulations, 2011 CFR

2011-07-01

... noncontaminating fittings are needed. For the first, third, and fourth impingers, use the Greenburg-Smith design....) from the bottom of the flask. For the second impinger, use the Greenburg-Smith design with the standard...

40 CFR Appendix A-5 to Part 60 - Test Methods 11 through 15A

Code of Federal Regulations, 2010 CFR

2010-07-01

... noncontaminating fittings are needed. For the first, third, and fourth impingers, use the Greenburg-Smith design....) from the bottom of the flask. For the second impinger, use the Greenburg-Smith design with the standard...

40 CFR Appendix A-5 to Part 60 - Test Methods 11 through 15A

Code of Federal Regulations, 2012 CFR

2012-07-01

... noncontaminating fittings are needed. For the first, third, and fourth impingers, use the Greenburg-Smith design....) from the bottom of the flask. For the second impinger, use the Greenburg-Smith design with the standard...

40 CFR Appendix A-5 to Part 60 - Test Methods 11 through 15A

Code of Federal Regulations, 2013 CFR

2013-07-01

... noncontaminating fittings are needed. For the first, third, and fourth impingers, use the Greenburg-Smith design....) from the bottom of the flask. For the second impinger, use the Greenburg-Smith design with the standard...

76 FR 53678 - Notice of Intent To Suspend Certain Pesticide Registrations

Federal Register 2010, 2011, 2012, 2013, 2014

2011-08-29

...- 12/14/2007 12/24/2007 8/20/2008 No data received. octanol/water) shake flask method. 19713-289 830.7570 Partition coefficient (n- 12/14/2007 12/24/2007 8/20/2008 No data received. octanol/water...

77 FR 10520 - Notice of Intent To Suspend Certain Pesticide Registrations

Federal Register 2010, 2011, 2012, 2013, 2014

2012-02-22

.../2007 12/24/2007 8/20/2008 No data coefficient (n- received. octanol/water) shake flask method. 19713-72......... 830.7570 Partition 12/14/2007 12/24/2007 8/20/2008 No data coefficient (n- received. octanol/water...

BIODEGRADATION OF FLUORANTHENE AS MONITORED USING STABLE CARBON ISOTOPES

EPA Science Inventory

The measurement of stable isotope ratios of carbon (d13C values) was investigated as a viable technique to monitor the intrinsic bioremediation of polycyclic aromatic hydrocarbons (PAHs). Biometer-flask experiments were conducted in which the bacterium, Sphingomonas paucimobilis,...

Improved Segmented-Flow Tracer-Monitored Titration for Automated Measurement of Total Alkalinity in Seawater

NASA Astrophysics Data System (ADS)

Spaulding, R. S.; Hales, B.; Beck, J. C.; Degrandpre, M. D.

2008-12-01

The four measurable inorganic carbon parameters commonly measured as part of oceanic carbon cycle studies are total dissolved inorganic carbon (DIC), total alkalinity (AT), hydrogen ion concentration (pH) and partial pressure of CO2 (pCO2). AT determination is critical for anthropogenic CO2 inventory calculations and for quantifying CaCO3 saturation. Additionally, measurement of AT in combination with one other carbonate parameter can be used to describe the inorganic carbon equilibria. Current methods for measuring AT require calibrated volumetric flasks and burettes, gravimetry, or precise flow measurements. These methods also require analysis times of ˜15 min and sample volumes of ˜200 mL, and sample introduction is not automated, resulting in labor-intensive measurements and low temporal resolution. The Tracer Monitored Titration (TMT) system was previously developed at the University of Montana for AT measurements. The TMT is not dependent on accurate gravimetric, volumetric or flow rate measurements because it relies on a pH-sensitive indicator (tracer) to track the amount of titrant added to the sample. Sample and a titrant-indicator mixture are mechanically stirred in an optical flow cell and pH is calculated using the indicator equilibrium constant and the spectrophotometrically determined concentrations of the acid and base forms of the indicator. AT is then determined using these data in a non-linear least squares regression of the AT mass and proton balances. The precision and accuracy of the TMT are 2 and 4 micromol per kg in 16 min using 110-mL of sample. The TMT is dependent on complete mixing of titrant with the sample and accurate absorbance measurements. We have developed the segmented-flow TMT (SF- TMT) to improve on these aspects and decrease sample analysis time. The TMT uses segmented flow instead of active mixing and a white LED instead of a tungsten-halogen light source. Air is added to the liquid flow stream, producing segments of liquid separated by air bubbles. Because liquid is not transferred between flow segments, there is rapid flushing which reduces sample volume to <10 mL. Additionally, the slower movement of liquid at the tube walls compared to that at the tube center creates circulation within each liquid segment, mixing the sample and eliminating the need for mechanical stirring. The white LED has higher output at the wavelengths of interest, thus improving the precision of absorbance measurements. These improvements result in a faster, simpler method for measuring AT.

75 FR 29540 - Notice of Suspension of Certain Pesticide Registrations

Federal Register 2010, 2011, 2012, 2013, 2014

2010-05-26

... (n- 59820-5 Octanol/H2O), Shake Flask Method 6297-6 830.7570 Partition August 15, 2008 August 21, 2008 April 30, 2009 No data received 59820-4 Coefficient (n- 59820-5 Octanol/H2O), Estimation by Liquid...

Streamlined bioreactor-based production of human cartilage tissues.

PubMed

Tonnarelli, B; Santoro, R; Adelaide Asnaghi, M; Wendt, D

2016-05-27

Engineered tissue grafts have been manufactured using methods based predominantly on traditional labour-intensive manual benchtop techniques. These methods impart significant regulatory and economic challenges, hindering the successful translation of engineered tissue products to the clinic. Alternatively, bioreactor-based production systems have the potential to overcome such limitations. In this work, we present an innovative manufacturing approach to engineer cartilage tissue within a single bioreactor system, starting from freshly isolated human primary chondrocytes, through the generation of cartilaginous tissue grafts. The limited number of primary chondrocytes that can be isolated from a small clinically-sized cartilage biopsy could be seeded and extensively expanded directly within a 3D scaffold in our perfusion bioreactor (5.4 ± 0.9 doublings in 2 weeks), bypassing conventional 2D expansion in flasks. Chondrocytes expanded in 3D scaffolds better maintained a chondrogenic phenotype than chondrocytes expanded on plastic flasks (collagen type II mRNA, 18-fold; Sox-9, 11-fold). After this "3D expansion" phase, bioreactor culture conditions were changed to subsequently support chondrogenic differentiation for two weeks. Engineered tissues based on 3D-expanded chondrocytes were more cartilaginous than tissues generated from chondrocytes previously expanded in flasks. We then demonstrated that this streamlined bioreactor-based process could be adapted to effectively generate up-scaled cartilage grafts in a size with clinical relevance (50 mm diameter). Streamlined and robust tissue engineering processes, as the one described here, may be key for the future manufacturing of grafts for clinical applications, as they facilitate the establishment of compact and closed bioreactor-based production systems, with minimal automation requirements, lower operating costs, and increased compliance to regulatory guidelines.

Catalytic Palladium Film Deposited by Scalable Low-Temperature Aqueous Combustion.

PubMed

Voskanyan, Albert A; Li, Chi-Ying Vanessa; Chan, Kwong-Yu

2017-09-27

This article describes a novel method for depositing a dense, high quality palladium thin film via a one-step aqueous combustion process which can be easily scaled up. Film deposition of Pd from aqueous solutions by conventional chemical or electrochemical methods is inhibited by hydrogen embrittlement, thus resulting in a brittle palladium film. The method outlined in this work allows a direct aqueous solution deposition of a mirror-bright, durable Pd film on substrates including glass and glassy carbon. This simple procedure has many advantages including a very high deposition rate (>10 cm 2 min -1 ) and a relatively low deposition temperature (250 °C), which makes it suitable for large-scale industrial applications. Although preparation of various high-quality oxide films has been successfully accomplished via solution combustion synthesis (SCS) before, this article presents the first report on direct SCS production of a metallic film. The mechanism of Pd film formation is discussed with the identification of a complex formed between palladium nitrate and glycine at low temperature. The catalytic properties and stability of films are successfully tested in alcohol electrooxidation and electrochemical oxygen reduction reaction. It was observed that combustion deposited Pd film on a glassy carbon electrode showed excellent catalytic activity in ethanol oxidation without using any binder or additive. We also report for the first time the concept of a reusable "catalytic flask" as illustrated by the Suzuki-Miyaura cross-coupling reaction. The Pd film uniformly covers the inner walls of the flask and eliminates the catalyst separation step. We believe the innovative concept of a reusable catalytic flask is very promising and has the required features to become a commercial product in the future.

Significance of oxygen supply in jarosite biosynthesis promoted by Acidithiobacillus ferrooxidans.

PubMed

Hou, Qingjie; Fang, Di; Liang, Jianru; Zhou, Lixiang

2015-01-01

Jarosite [(Na+, K+, NH4+, H3O+)Fe3(SO4)2(OH)6] is an efficient scavenger for trace metals in Fe- and SO42--rich acidic water. During the biosynthesis of jarosite promoted by Acidithiobacillus ferrooxidans, the continuous supply of high oxygen levels is a common practice that results in high costs. To evaluate the function of oxygen in jarosite production by A. ferrooxidans, three groups of batch experiments with different oxygen supply levels (i.e., loading volume percentages of FeSO4 solution of 20%, 40%, and 70% v/v in the flasks), as well as three groups of sealed flask experiments with different limiting oxygen supply conditions (i.e., the solutions were not sealed at the initial stage of the ferrous oxidation reaction by paraffin but were rather sealed at the end of the ferrous oxidation reaction at 48 h), were tested. The formed Fe-precipitates were characterized via X-ray powder diffraction and scanning electron microscope-energy dispersive spectral analysis. The results showed that the biosynthesis of jarosite by A. ferrooxidans LX5 could be achieved at a wide range of solution loading volume percentages. The rate and efficiency of the jarosite biosynthesis were poorly correlated with the concentration of dissolved oxygen in the reaction solution. Similar jarosite precipitates, expressed as KFe3 (SO4) 2(OH)6 with Fe/S molar ratios between 1.61 and 1.68, were uniformly formed in unsealed and 48 h sealed flasks. These experimental results suggested that the supply of O2 was only essential in the period of the oxidation of ferrous iron to ferric but was not required in the period of ferric precipitation.

Significance of Oxygen Supply in Jarosite Biosynthesis Promoted by Acidithiobacillus ferrooxidans

PubMed Central

Liang, Jianru; Zhou, Lixiang

2015-01-01

Jarosite [(Na+, K+, NH4 +, H3O+)Fe3(SO4)2(OH)6] is an efficient scavenger for trace metals in Fe- and SO4 2--rich acidic water. During the biosynthesis of jarosite promoted by Acidithiobacillus ferrooxidans, the continuous supply of high oxygen levels is a common practice that results in high costs. To evaluate the function of oxygen in jarosite production by A. ferrooxidans, three groups of batch experiments with different oxygen supply levels (i.e., loading volume percentages of FeSO4 solution of 20%, 40%, and 70% v/v in the flasks), as well as three groups of sealed flask experiments with different limiting oxygen supply conditions (i.e., the solutions were not sealed at the initial stage of the ferrous oxidation reaction by paraffin but were rather sealed at the end of the ferrous oxidation reaction at 48 h), were tested. The formed Fe-precipitates were characterized via X-ray powder diffraction and scanning electron microscope-energy dispersive spectral analysis. The results showed that the biosynthesis of jarosite by A. ferrooxidans LX5 could be achieved at a wide range of solution loading volume percentages. The rate and efficiency of the jarosite biosynthesis were poorly correlated with the concentration of dissolved oxygen in the reaction solution. Similar jarosite precipitates, expressed as KFe3 (SO4) 2(OH)6 with Fe/S molar ratios between 1.61 and 1.68, were uniformly formed in unsealed and 48 h sealed flasks. These experimental results suggested that the supply of O2 was only essential in the period of the oxidation of ferrous iron to ferric but was not required in the period of ferric precipitation. PMID:25807372

Observation of development of breast cancer cell lines in real time by fluorescence microscopy under simulated microgravity

NASA Astrophysics Data System (ADS)

Lavan, David; Valdivia-Silva, Julio E.; Sanabria, Gabriela; Orihuela, Diego; Suarez, Juan; Quispe, Marco; Chuchon, Mariano; Martin, David; Maroto, Marcos; Egea, Javier

2016-07-01

This project consist in the implementation of a fluorescence microscope for the in real time monitoring of biological labeled samples by several fluorophores in microgravity conditions keeping the temperature, humidity, and (CO)2 controlled by an electronic platform. The system (fluorescence microscope and incubator) is integrated to a microgravity simulator machine which was presented on the "30th Annual American Society for Gravitation and Space Research Meeting" October 2014 in Pasadena, CA, USA. Currently, we have the microgravity machine biologically validated by genetic expression studies in pupal stage of Drosophila melanogaster. The fluorescence microscope has a platform designed to hold a culture flask, and a fluorescence camera (Leica DFC3000 G) connected to an optical system (Fluorescence Light source Leica EL6000, optic fiber, fiber adapter, and fluorescence filter) in order to take images in real time. The mechanical system of the fluorescence microsc ope is designed to allow the displacement of the fluorescence camera through a parallel plane to the culture flask's plane and also the movement of the platform through a perpendicular axis to the culture flask in order to focus the samples to the optical system. The mechanical system is propelled by four DC moto-reductors with encoder (A-max 26 Maxon motor, GP 32S screw and MR encoder) that generate displacements in the order of micrometers. The angular position control of the DC motoreductor's shaft of all the DC moto-reductors is done by PWM signals based on the interpretation of the signals provided by the encoders during the movement. The system is remotely operated by a graphic interface installed on a personal computer or any mobile device (smartphone, laptop or tablet) by using the internet. Acknowledgments: Grant of INNOVATE PERU (Formerly FINCYT)

Collection and Analysis of Firn Air from the South Pole, 2001

NASA Astrophysics Data System (ADS)

Butler, J. H.; Montzka, S. A.; Battle, M.; Clarke, A. D.; Mondeel, D. J.; Lind, J. A.; Hall, B. D.; Elkins, J. W.

2001-12-01

In January 2001, we collected an archive of 20th century air from the firn (snowpack) at the South Pole. Samples were collected into separate pairs of 3L glass flasks for measurements of O2/N2 (Bowdoin/Princeton) and carbon cycle gases (CMDL); individual 3L stainless steel and glass flasks for measurements of halocarbons, N2O, SF6, and COS; large (33L) stainless steel canisters for maintaining an archive of air for future analyses; and a few canisters each for measurement of 14CH4 (NIWA/CSIRO) and very low-level analyses of SF6 (SIO). Although it was hoped to obtain air dating back to the turn of the century, the analyses suggest that the earliest date was 1925 for CO2 and the mid- to late teens for heavier gases such as methyl bromide or methyl chloride. This talk will compare the analyses of halocarbons in these recently collected samples to those of air in flasks sampled at the South Pole in 1995. We also will present some results for compounds not measured in the 1995 South Pole samples owing to a paucity of air. Measurements made of the same gases in the firn air at both ends of this six-year interval, along with real-time atmospheric measurements of the same gases, are useful in evaluating assumptions about diffusion in the firn and may allow for the direct calculation of diffusion coefficients at low temperatures. This, in turn, would improve age estimates for firn air samples. New measurements will add to our existing histories established for the 20th century from analyses of firn air samples collected in both Greenland and Antarctica.

Onboard measurement system of atmospheric carbon monoxide in the Pacific by voluntary observing ships

NASA Astrophysics Data System (ADS)

Nara, H.; Tanimoto, H.; Nojiri, Y.; Mukai, H.; Machida, T.; Tohjima, Y.

2011-11-01

Long-term monitoring of carbon monoxide (CO) mixing ratios in the atmosphere over the Pacific Ocean is being carried out on commercial cargo vessels participating in the National Institute for Environmental Studies Voluntary Observing Ships program. The program provides a regular platform for measurement of atmospheric CO along four cruise routes: from Japan to Oceania, the United States, Canada, and Southeast Asia. Flask samples are collected during every cruise for subsequent analysis in the laboratory, and in 2005, continuous shipboard CO measurements were initiated on three of the routes. Here, we describe the system we developed for onboard measurement of CO mixing ratios with a commercially available gas filter correlation CO analyzer. The fully automated system measures CO in ambient air, and the detector sensitivity and background signals are calibrated by referencing the measurements to a CO-in-air standard gas (~1 ppmv) and to CO-free air scrubbed with a catalyst, respectively. We examined the artificial production of CO in the high-pressure working gas standards during storage by referencing the measurements to CO standard gases maintained as our primary scale before and after use on the ships. The onboard performance of the continuous CO measurement system was evaluated by comparing its data with data from laboratory analyses of flask samples using gas chromatography with a reduction gas detector. The reasonably good consistency between the two independent measurement methods demonstrated the good performance of both methods over the course of 3-5 years. The continuous measurement system was more useful than the flask sampling method for regionally polluted air masses, which were often encountered on Southeast Asian cruises.

Onboard measurement system of atmospheric carbon monoxide over the Pacific Ocean by voluntary observing ships

NASA Astrophysics Data System (ADS)

Nara, H.; Tanimoto, H.; Nojiri, Y.; Mukai, H.; Machida, T.; Tohjima, Y.

2011-07-01

Long-term monitoring of carbon monoxide (CO) mixing ratios in the atmosphere over the Pacific Ocean is being carried out on commercial cargo vessels participating in the National Institute for Environmental Studies Voluntary Observing Ships program. The program provides a regular platform for measurement of atmospheric CO along four cruising routes: from Japan to Oceania, from Japan to the United States, from Japan to Canada, and from Japan to Southeast Asia. Flask samples are collected during every cruise for subsequent analysis in the laboratory, and in 2005, continuous shipboard CO measurements were initiated on three of the routes. Here, we describe the system we developed for onboard measurement of CO mixing ratios with a commercially available gas filter correlation CO analyzer. The fully automated system measures CO in ambient air, and the detector sensitivity and background signals are calibrated by referencing the measurements to a CO-in-air standard gas (~1 ppmv) and to CO-free air scrubbed with a catalyst, respectively. We examined the artificial production of CO in the high-pressure working gas standards (CO balanced with purified air at ppmv levels) during storage by referencing the measurements to CO standard gases maintained as our primary scale before and after use on the ships. The onboard performance of the continuous CO measurement system was evaluated by comparing its data with data from laboratory analyses of flask samples using gas chromatography with a reduction gas detector. The reasonably good consistency between the two independent measurement methods demonstrated the good performance of both methods over the course of 3-5 yr. The continuous measurement system was more useful than the flask sampling method for regionally polluted air masses, which were often encountered on Southeast Asian cruises.

21 CFR 177.1970 - Vinyl chloride-lauryl vinyl ether copolymers.

Code of Federal Regulations, 2012 CFR

2012-04-01

... filter paper with the aid of suction. Transfer the filtrate to flat glass dishes that are warmed on a hot.... Maintain the temperature of the contents of the flask at 150 °F for 2 hours using a hot plate while also...

21 CFR 177.1970 - Vinyl chloride-lauryl vinyl ether copolymers.

Code of Federal Regulations, 2011 CFR

2011-04-01

... filter paper with the aid of suction. Transfer the filtrate to flat glass dishes that are warmed on a hot.... Maintain the temperature of the contents of the flask at 150 °F for 2 hours using a hot plate while also...

21 CFR 177.1970 - Vinyl chloride-lauryl vinyl ether copolymers.

Code of Federal Regulations, 2013 CFR

2013-04-01

... filter paper with the aid of suction. Transfer the filtrate to flat glass dishes that are warmed on a hot.... Maintain the temperature of the contents of the flask at 150 °F for 2 hours using a hot plate while also...

21 CFR 177.1970 - Vinyl chloride-lauryl vinyl ether copolymers.

Code of Federal Regulations, 2010 CFR

2010-04-01

... filter paper with the aid of suction. Transfer the filtrate to flat glass dishes that are warmed on a hot.... Maintain the temperature of the contents of the flask at 150 °F for 2 hours using a hot plate while also...

49 CFR 173.320 - Cryogenic liquids; exceptions.

Code of Federal Regulations, 2010 CFR

2010-10-01

... atmospheric gases and helium: (1) During loading and unloading operations (pressure rises may exceed 25.3 psig...; exceptions. (a) Atmospheric gases and helium, cryogenic liquids, in Dewar flasks, insulated cylinders... the pressure in such packagings will not exceed 25.3 psig under ambient temperature conditions during...

41. #1 ARRESTING GEAR ENGINE AFT LOOKING FORWARD PORT ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

41. #1 ARRESTING GEAR ENGINE - AFT LOOKING FORWARD PORT TO STARBOARD SHOWING ARRESTING GEAR ENGINE ACCUMULATOR, AIR FLASK, CONTROL VALVE, WITH CONTROL RAM, SHEAVES AND WIRES UNDERNEATH ENGINE STAND. - U.S.S. HORNET, Puget Sound Naval Shipyard, Sinclair Inlet, Bremerton, Kitsap County, WA

OIL SPILL DISPERSANT EFFECTIVENESS PROTOCOL. I: IMPACT OF OPERATIONAL VARIABLES

EPA Science Inventory

The current U.S. Environmental Protection Agency protocol for testing the effectiveness of dispersants, the swirling flask test, has been found to give widely varying results in the hands of different testing laboratories. The sources of the ambiguities in the test were determin...

Identifying and overcoming the effect of mass transfer limitation on decreased yield in enzymatic hydrolysis of lignocellulose at high solid concentrations.

PubMed

Du, Jian; Cao, Yuan; Liu, Guodong; Zhao, Jian; Li, Xuezhi; Qu, Yinbo

2017-04-01

Cellulose conversion decreases significantly with increasing solid concentrations during enzymatic hydrolysis of insoluble lignocellulosic materials. Here, mass transfer limitation was identified as a significant determining factor of this decrease by studying the hydrolysis of delignified corncob residue in shake flask, the most used reaction vessel in bench scale. Two mass transfer efficiency-related factors, mixing speed and flask filling, were shown to correlate closely with cellulose conversion at solid loadings higher than 15% DM. The role of substrate characteristics in mass transfer performance was also significant, which was revealed by the saccharification of two corn stover substrates with different pretreatment methods at the same solid loading. Several approaches including premix, fed-batch operation, and particularly the use of horizontal rotating reactor were shown to be valid in facilitating cellulose conversion via improving mass transfer efficiency at solid concentrations higher than 15% DM. Copyright © 2017 Elsevier Ltd. All rights reserved.

Production of poly-beta-hydroxybutyric acid by microorganisms accumulated from river water using a two-stage perfusion culture system.

PubMed

Morimoto, T; Tashiro, F; Nagashima, H; Nishizawa, K; Nagata, F; Yokogawa, Y; Suzuki, T

2000-01-01

The perfusion culture system using a shaken ceramic membrane flask (SCMF) was employed to accumulate microorganisms separated from river water and to produce poly-beta-hydroxybutyric acid (PHB). Using a two-step culture method with a single SCMF, river microorganisms were cultured by separately feeding four representative carbon sources, n-propanol, lactic acid, methanol, and formic acid. After 140 h culture, the cell concentration and PHB content respectively reached 43 g/l and 35% when a propanol medium was fed. Using a two-stage perfusion culture with twin SCMFs, the seed cell mass was increased in the first SCMF and then supplied to the second flask for PHB production. As a consequence, the cellular PHB content rose to 51% in the second SCMF, while the cell concentration gradually increased to 25 g/l after 175 h perfusion culture. These results demonstrated the utility of the two-stage perfusion culture system for developing a cheap means of producing PHB coincident with wastewater treatment.

[Enhanced ε-poly-L-lysine production by improving cellular activity during fermentation].

PubMed

Liu, Shengrong; Wu, Qingping; Zhang, Jumei; Yang, Xiaojuan; Cai, Shuzhen

2015-06-04

To assess the effect of cellular activity on ε-poly-1-lysine (ε-PL) biosynthesis and thereby to rationally improve the production, we studied the cellular activity, ε-PL formation and other parameters cross flask fermentation by Streptomyces ahygroscopicus. Laser scanning confocal microscopy and a colorimetric method were used to determine cellular activity using BacLight Live/Dead and 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) as viable stains. To enhance the activity of the cells in the ε-PL production period, yeast extract was added. During ε-PL submerged fermentation in flasks, most cells were active in the growth period (0 - 16 h); cells had metabolic activity in the growth and earlier ε-PL production periods between 0 and 30 h fermentation. Almost no activity was detected after 48 h fermentation when no ε-PL was produced. The improved fermentation achieved 2. 24 g/L ε-PL from 1.04 g/L. Biosynthesis of ε-PL can be boosted by up-regulating cell activity in its production phase.

Cost analysis in laccase production.

PubMed

Osma, Johann F; Toca-Herrera, José L; Rodríguez-Couto, Susana

2011-11-01

In this paper the cost of producing the enzyme laccase by the white-rot fungus Trametes pubescens under both submerged (SmF) and solid-state fermentation (SSF) conditions was studied. The fungus was cultured using more than 45 culture medium compositions. The cost of production was estimated by analyzing the cost of the culture medium, the cost of equipment and the operating costs. The cost of the culture medium represented, in all cases, the highest contribution to the total cost, while, the cost of equipment was significantly low, representing less than 2% of the total costs. The cultivation under SSF conditions presented a final cost 50-fold lower than the one obtained when culturing under SmF conditions at flask scale. In addition, the laccase production under SSF conditions in tray bioreactors reduced the final cost 4-fold compared to the one obtained under SSF conditions at flask scale, obtaining a final price of 0.04 cent €/U. Copyright © 2011 Elsevier Ltd. All rights reserved.

Expansion of Human Mesenchymal Stem Cells in a Microcarrier Bioreactor.

PubMed

Tsai, Ang-Chen; Ma, Teng

2016-01-01

Human mesenchymal stem cells (hMSCs) are considered as a primary candidate in cell therapy owing to their self-renewability, high differentiation capabilities, and secretions of trophic factors. In clinical application, a large quantity of therapeutically competent hMSCs is required that cannot be produced in conventional petri dish culture. Bioreactors are scalable and have the capacity to meet the production demand. Microcarrier suspension culture in stirred-tank bioreactors is the most widely used method to expand anchorage dependent cells in a large scale. Stirred-tank bioreactors have the potential to scale up and microcarriers provide the high surface-volume ratio. As a result, a spinner flask bioreactor with microcarriers has been commonly used in large scale expansion of adherent cells. This chapter describes a detailed culture protocol for hMSC expansion in a 125 mL spinner flask using microcarriers, Cytodex I, and a procedure for cell seeding, expansion, metabolic sampling, and quantification and visualization using microculture tetrazolium (MTT) reagent.

Aeolian comminution experiments revealing surprising sandball mineral aggregates

NASA Astrophysics Data System (ADS)

Nørnberg, P.; Bak, E.; Finster, K.; Gunnlaugsson, H. P.; Iversen, J. J.; Jensen, S. Knak; Merrison, J. P.

2014-06-01

We have undertaken a set of wind erosion experiments on a simple and well defined mineral, quartz. In these experiments wind action is simulated by end over end tumbling of quartz grains in a sealed quartz flask. The tumbling induces collisions among the quartz grains and the walls of the flask. This process simulates wind action impact speed of ∼1.2 m/s. After several months of tumbling we observed the formation of a large number of spherical sand aggregates, which resemble small snowballs under optical microscopy. Upon mechanical load the aggregates are seen to be more elastic than quartz and their mechanical strength is comparable, though slightly lower than that of sintered silica aerogels. Aggregates of this kind have not been reported from field sites or from closed circulation systems. However, sparse occurrence might explain this, or in nature the concentration of the aggregate building particles is so low that they never meet and just appear as the most fine grained tail of the sediment particle size distribution.

Simultaneous treatment of food-waste recycling wastewater and cultivation of Tetraselmis suecica for biodiesel production.

PubMed

Heo, Sung-Woon; Ryu, Byung-Gon; Nam, Kibok; Kim, Woong; Yang, Ji-Won

2015-07-01

There is an increasing interest in the use of cultivated microalgae to simultaneously produce biodiesel and remove nutrients from various wastewaters. For this purpose, Tetraselmis suecica was cultivated in flasks and fermenters using diluted food-waste recycling wastewater (FRW). The effect of FRW dilution on T. suecica growth and nutrient removal was initially tested in flasks. The maximal microalgal concentration after 14 days was in medium with a twofold dilution (28.3 × 10(6) cells/mL) and a fivefold dilution (25.5 × 10(6) cells/mL). When fivefold diluted medium was used in fermenters, the final dry cell weight of T. suecica was 2.0 g/L. The removal efficiencies of ammonium and phosphate in the fermenters were 99.0 and 52.3%, respectively. In comparison with the results of previous studies, the growth data of T. suecica in the FRW medium indicate that microalgal cultivation system incorporating removal of nutrients in FRW is feasible at the field level.

QSAR models for predicting octanol/water and organic carbon/water partition coefficients of polychlorinated biphenyls.

PubMed

Yu, S; Gao, S; Gan, Y; Zhang, Y; Ruan, X; Wang, Y; Yang, L; Shi, J

2016-04-01

Quantitative structure-property relationship modelling can be a valuable alternative method to replace or reduce experimental testing. In particular, some endpoints such as octanol-water (KOW) and organic carbon-water (KOC) partition coefficients of polychlorinated biphenyls (PCBs) are easier to predict and various models have been already developed. In this paper, two different methods, which are multiple linear regression based on the descriptors generated using Dragon software and hologram quantitative structure-activity relationships, were employed to predict suspended particulate matter (SPM) derived log KOC and generator column, shake flask and slow stirring method derived log KOW values of 209 PCBs. The predictive ability of the derived models was validated using a test set. The performances of all these models were compared with EPI Suite™ software. The results indicated that the proposed models were robust and satisfactory, and could provide feasible and promising tools for the rapid assessment of the SPM derived log KOC and generator column, shake flask and slow stirring method derived log KOW values of PCBs.

Characterization of a thermostable raw-starch hydrolyzing α-amylase from deep-sea thermophile Geobacillus sp.

PubMed

Jiang, Tao; Cai, Menghao; Huang, Mengmeng; He, Hao; Lu, Jian; Zhou, Xiangshan; Zhang, Yuanxing

2015-10-01

A deep-sea thermophile, Geobacillus sp. 4j, was identified to grow on starch and produce thermostable amylase. N-terminally truncated form of Geobacillus sp. 4j α-amylase (Gs4j-amyA) was fused at its N-terminal end with the signal peptide of outer membrane protein A (OmpA) of Escherichia coli. The enzyme was over-expressed in E. coli BL21 with a maximum extracellular production of 130U/ml in shake flask. The yield of the transformant increased 22-fold as compared with that of the wild strain. The recombinant enzyme purified to apparent homogeneity by metal-affinity chromatography, exhibited a molecular mass of 62kDa. It displayed the maximal activity at 60-65°C and pH 5.5. Its half-life (t1/2) at 80°C was 4.25h with a temperature deactivation energy of 166.3kJ/mol. Compared to three commonly used commercial α-amylases, the Gs4j-amyA exhibited similar thermostable performance to BLA but better than BAA and BSA. It also showed a universally efficient raw starch hydrolysis performance superior to commercial α-amylases at an acidic pH approaching nature of starch slurry. As a new acidic-resistant thermostable α-amylase, it has the potential to bypass the industrial gelatinization step in raw starch hydrolysis. Copyright © 2015 Elsevier Inc. All rights reserved.

Enhanced production of acarbose and concurrently reduced formation of impurity c by addition of validamine in fermentation of Actinoplanes utahensis ZJB-08196.

PubMed

Xue, Ya-Ping; Qin, Jun-Wei; Wang, Ya-Jun; Wang, Yuan-Shan; Zheng, Yu-Guo

2013-01-01

Commercial production of acarbose is exclusively via done microbial fermentation with strains from the genera of Actinoplanes. The addition of C7N-aminocyclitols for enhanced production of acarbose and concurrently reduced formation of impurity C by cultivation of A. utahensis ZJB-08196 in 500-mL shake flasks was investigated, and validamine was found to be the most effective strategy. Under the optimal conditions of validamine addition, acarbose titer was increased from 3560 ± 128 mg/L to 4950 ± 156 mg/L, and impurity C concentration was concurrently decreased from 289 ± 24 mg/L to 107 ± 29 mg/L in batch fermentation after 168 h of cultivation. A further fed-batch experiment coupled with the addition of validamine (20 mg/L) in the fermentation medium prior to inoculation was designed to enhance the production of acarbose. When twice feedings of a mixture of 6 g/L glucose, 14 g/L maltose, and 9 g/L soybean flour were performed at 72 h and 96 h, acarbose titer reached 6606 ± 103 mg/L and impurity C concentration was only 212 ± 12 mg/L at 168 h of cultivation. Acarbose titer and proportion of acarbose/impurity C increased by 85.6% and 152.9% when compared with control experiments. This work demonstrates for the first time that validamine addition is a simple and effective strategy for increasing acarbose production and reducing impurity C formation.

Molecular cloning and characterization of (+)-epi-α-bisabolol synthase, catalyzing the first step in the biosynthesis of the natural sweetener, hernandulcin, in Lippia dulcis.

PubMed

Attia, Mohamed; Kim, Soo-Un; Ro, Dae-Kyun

2012-11-01

Hernandulcin, a C15 sesquiterpene ketone, is a natural sweetener isolated from the leaves of Lippia dulcis. It is a promising sugar substitute due to its safety and low caloric potential. However, the biosynthesis of hernandulcin in L. dulcis remains unknown. The first biochemical step of hernandulcin is the synthesis of (+)-epi-α-bisabolol from farnesyl diphosphate, which is presumed to be catalyzed by a unique sesquiterpene synthase in L. dulcis. In order to decipher hernandulcin biosynthesis, deep transcript sequencings (454 and Illumina) were performed, which facilitated the molecular cloning of five new sesquiterpene synthase cDNAs from L. dulcis. In vivo activity evaluation of these cDNAs in yeast identified them as the sesquiterpene synthases for α-copaene/δ-cadinene, bicyclogermacrene, β-caryophyllene, trans-α-bergamotene, and α-bisabolol. The engineered yeast could synthesize a significant amount (~0.3 mg per mL) of α-bisabolol in shake-flask cultivation. This efficient in vivo production was congruent with the competent kinetic properties of recombinant α-bisabolol synthase (K(m) 4.8 μM and k(cat) 0.04 s(-1)). Detailed chemical analyses of the biosynthesized α-bisabolol confirmed its configuration to be (+)-epi-α-bisabolol, the core skeleton of hernandulcin. These results demonstrated that enzymatic, stereoselective synthesis of (+)-epi-α-bisabolol can be achieved, promising the heterologous production of a natural sweetener, hernandulcin. Copyright © 2012 Elsevier Inc. All rights reserved.

Analysis of proteins involved in biodegradation of crop biomass

NASA Technical Reports Server (NTRS)

Crawford, Kamau; Trotman, Audrey

1998-01-01

The biodegradation of crop biomass for re-use in crop production is part of the bioregenerative life support concept proposed by the National Aeronautics and Space Administration (NASA) for long duration, manned space exploration. The current research was conducted in the laboratory to evaluate the use of electrophoretic analysis as a means of rapidly assaying for constitutive and induced proteins associated with the bacterial degradation of crop residue. The proteins involved in crop biomass biodegradation are either constitutive or induced. As a result, effluent and cultures were examined to investigate the potential of using electrophoretic techniques as a means of monitoring the biodegradation process. Protein concentration for optimum banding patterns was determined using the Bio-Rad Protein Assay kit. Four bacterial soil isolates were obtained from the G.W. Carver research Farm at Tuskegee University and used in the decomposition of components of plant biomass. The culture, WDSt3A was inoculated into 500 mL of either Tryptic Soy Broth or Nutrient Broth. Incubation, with shaking of each flask was for 96 hours at 30 C. The cultures consistently gave unique banding patterns under denaturing protein electrophoresis conditions, The associated extracellular enzymes also yielded characteristic banding patterns over a 14-day period, when native electrophoresis techniques were used to examine effluent from batch culture bioreactors. The current study evaluated sample preparation and staining protocols to determine the ease of use, reproducibility and reliability, as well as the potential for automation.

PRODUCTION AND CHARACTERIZATION OF MULTIPLE-LAYERED POPULATIONS OF ANIMAL CELLS

PubMed Central

Kruse, Paul F.; Miedema, Ed

1965-01-01

Dense populations containing 129 x 106 Jensen sarcoma, 134 x 106 DON Chinese hamster, 28.9 x 106 WI-38 human diploid, 61.8 x 106 HEp-2 human carcinoma, and 67.4 x 106 WISH human amnion cells were produced from dilute inocula, 0.85 to 5.33 x 106, in 7 to 8 days in a perfusion system using replicate T-60 flasks. Perfusion rates as high as 560 ml medium/day/T-60 were required to maintain pH (to ca ±0.1 unit) and adequate nutrient supplies. The cell densities encountered are described by the term "monolayer equivalents" (M.E.), defined as number of cells per culture divided by number of cells in a monolayer. The M.E.'s for T-60 cultures containing unusually dense populations of 40 x 106 WI-38 and 250 x 106 DON cells (9-day perfusion) were 5 and 17, respectively, and numbers of cells in illustrations of stained cross-sections of membranes from these cultures were in excellent agreement. Threshold M.E.'s exist below which proliferation is the chief cellular activity and above which one or more cell functions may predominate even though proliferation persists. Cellular nutrition and metabolism may change with changes in M.E., as illustrated in different patterns of glutamic acid, proline, and glycine utilization or production in dense vs. dilute WI-38 cell populations. The results indicated that the role of contact inhibition phenomena in arresting cellular proliferation was diminished in perfusion system environments. PMID:5884626

A novel wild-type Saccharomyces cerevisiae strain TSH1 in scaling-up of solid-state fermentation of ethanol from sweet sorghum stalks.

PubMed

Du, Ran; Yan, Jianbin; Feng, Quanzhou; Li, Peipei; Zhang, Lei; Chang, Sandra; Li, Shizhong

2014-01-01

The rising demand for bioethanol, the most common alternative to petroleum-derived fuel used worldwide, has encouraged a feedstock shift to non-food crops to reduce the competition for resources between food and energy production. Sweet sorghum has become one of the most promising non-food energy crops because of its high output and strong adaptive ability. However, the means by which sweet sorghum stalks can be cost-effectively utilized for ethanol fermentation in large-scale industrial production and commercialization remains unclear. In this study, we identified a novel Saccharomyces cerevisiae strain, TSH1, from the soil in which sweet sorghum stalks were stored. This strain exhibited excellent ethanol fermentative capacity and ability to withstand stressful solid-state fermentation conditions. Furthermore, we gradually scaled up from a 500-mL flask to a 127-m3 rotary-drum fermenter and eventually constructed a 550-m3 rotary-drum fermentation system to establish an efficient industrial fermentation platform based on TSH1. The batch fermentations were completed in less than 20 hours, with up to 96 tons of crushed sweet sorghum stalks in the 550-m3 fermenter reaching 88% of relative theoretical ethanol yield (RTEY). These results collectively demonstrate that ethanol solid-state fermentation technology can be a highly efficient and low-cost solution for utilizing sweet sorghum, providing a feasible and economical means of developing non-food bioethanol.

Thermoelectrically cooled temperature-gradient apparatus for comparative cell and virus temperature studies.

PubMed

Clark, H F; Kaminski, F; Karzon, D T

1970-05-01

Establishment of a near-linear temperature gradient in an incubator has been accomplished by the application of heat to one terminus of a conducting body, normally a metal bar, and the removal of heat from the other terminus of the conducting body. Such incubators have been complex and unwieldy because of the need for mechanical refrigeration. We have described a simplified temperature gradient incubator which uses thermoelectric module cooling coupled with electric heating. Along the gradient, 20 stations in two parallel rows of 10, each accommodating a 30-ml plastic cell culture flask, were continually monitored by an electronic thermometer, and the temperatures were recorded. By manipulation of two simple potentiometer controls, any temperature gradient between 0 and 50 C could be obtained. Minor deviations which occurred between theoretically perfect and obtained temperature gradients were reproducible and readily measured. The gradient incubator was particularly applicable to (i) simultaneously studying a given biological activity over the entire temperature range supporting the growth of a given cell, virus, or microorganism, or (ii) precisely defining the upper or lower temperature limits of a biological system by 10-point determinations. Preliminary experiments have demonstrated the usefulness of the apparatus in characterizing the temperature limits for growth in vitro of cells of reptilian cell lines. The gradient incubator was also successfully utilized for the characterization of the effect of temperature on the efficiency of plating of amphibian viruses and possible temperature variants of those viruses.

A Novel Wild-Type Saccharomyces cerevisiae Strain TSH1 in Scaling-Up of Solid-State Fermentation of Ethanol from Sweet Sorghum Stalks

PubMed Central

Feng, Quanzhou; Li, Peipei; Zhang, Lei; Chang, Sandra; Li, Shizhong

2014-01-01

The rising demand for bioethanol, the most common alternative to petroleum-derived fuel used worldwide, has encouraged a feedstock shift to non-food crops to reduce the competition for resources between food and energy production. Sweet sorghum has become one of the most promising non-food energy crops because of its high output and strong adaptive ability. However, the means by which sweet sorghum stalks can be cost-effectively utilized for ethanol fermentation in large-scale industrial production and commercialization remains unclear. In this study, we identified a novel Saccharomyces cerevisiae strain, TSH1, from the soil in which sweet sorghum stalks were stored. This strain exhibited excellent ethanol fermentative capacity and ability to withstand stressful solid-state fermentation conditions. Furthermore, we gradually scaled up from a 500-mL flask to a 127-m3 rotary-drum fermenter and eventually constructed a 550-m3 rotary-drum fermentation system to establish an efficient industrial fermentation platform based on TSH1. The batch fermentations were completed in less than 20 hours, with up to 96 tons of crushed sweet sorghum stalks in the 550-m3 fermenter reaching 88% of relative theoretical ethanol yield (RTEY). These results collectively demonstrate that ethanol solid-state fermentation technology can be a highly efficient and low-cost solution for utilizing sweet sorghum, providing a feasible and economical means of developing non-food bioethanol. PMID:24736641

Synbiotic functional drink from Jerusalem artichoke juice fermented by probiotic Lactobacillus plantarum PCS26.

PubMed

Dimitrovski, Darko; Velickova, Elena; Dimitrovska, Maja; Langerholc, Tomaz; Winkelhausen, Eleonora

2016-01-01

A probiotic strain Lactobacillus plantarum PCS26 was used to ferment Jerusalem artichoke juice. Growth kinetics of the bacterial strain was followed during juice fermentation both in flask and in laboratory fermentor. Jerusalem artichoke showed to be an excellent source of nutrients for L. plantarum PCS26 growth. The culture grew very well reaching more than 10(10) cfu/ml in just 12 h. The pH changed from the initial 6.5 to 4.6 at the end of fermentation. The culture hydrolyzed fructooligosaccharides present in the Jerusalem artichoke juice, yielding fructose which was presumably consumed along with the malic acid as energy and carbon source. Lactic acid was the main metabolite produced in concentration of 4.6 g/L. Acetic and succinic acid were also identified. Sensory evaluation of the fermented Jerusalem artichoke juice and its mixtures with blueberry juice showed that the 50/50 % v/v mixture would be very well accepted by the consumers. Above 80 % of the panelists would buy this drink, and over 60 % were willing to pay more for it. Culture survivability in the fermented juices during storage at 4-7 °C was assayed by the Weibullian model. The product shelf-life was extended from 19.70 ± 0.50 days of pure Jerusalem artichoke juice to 35.7 ± 6.4 days of the mixture containing 30 % blueberry juice.

Canine intestinal contents vs. simulated media for the assessment of solubility of two weak bases in the human small intestinal contents.

PubMed

Kalantzi, Lida; Persson, Eva; Polentarutti, Britta; Abrahamsson, Bertil; Goumas, Konstantinos; Dressman, Jennifer B; Reppas, Christos

2006-06-01

This study was conducted to assess the relative usefulness of canine intestinal contents and simulated media in the prediction of solubility of two weak bases (dipyridamole and ketoconazole) in fasted and fed human intestinal aspirates that were collected under conditions simulating those in bioavailability/bioequivalence studies. After administration of 250 mL of water or 500 mL of Ensure plus [both containing 10 mg/mL polyethylene glycol (PEG) 4000 as nonabsorbable marker], intestinal aspirates were collected from the fourth part of the duodenum of 12 healthy adults and from the mid-jejunum of four Labradors. Pooled samples were analyzed for PEG, pH, buffer capacity, osmolality, surface tension, pepsin, total carbohydrates, total protein content, bile salts, phospholipids, and neutral lipids. The shake-flask method was used to measure the solubility of dipyridamole and ketoconazole in pooled human and canine intestinal contents and in fasted-state-simulating intestinal fluid (FaSSIF) and fed-state-simulating intestinal fluid (FeSSIF) containing various bile salts and pH-buffering agents. For both compounds, solubility in canine contents may be predictive of human intralumenal solubility in the fasting state but not in the fed state. The poor agreement of results in canine and human aspirates can be attributed to the higher bile salt content in canine bile. Solubility in FaSSIF containing a mixture of bile salts from crude bile predicted satisfactorily the intralumenal solubility of both drugs in the fasted state in humans. Solubility in FeSSIF, regardless of the identity of bile salts or of the buffering species, deviated from intralumenal values in the fed human aspirates by up to 40%. This was attributed to the lack of lipolytic products in FeSSIF, the higher bile salt content of FeSSIF, and the lower pH of FeSSIF. FaSSIF containing a mixture of bile salts from crude bile, and FeSSIF containing lipolytic products and, perhaps, having lower bile salt content but slightly higher pH, should be more useful than canine intestinal aspirates for predicting intralumenal solubilities in humans.

A two-stage refinement approach for the enhancement of excretory production of an exoglucanase from Escherichia coli.

PubMed

Fu, Z B; Ng, K L; Lam, C C; Leung, K C; Yip, W H; Wong, W K R

2006-08-01

Hyper-expression of a secretory exoglucanase, Exg, encoded by the cex gene of Cellulomonas fimi was previously shown to saturate the SecYEG pathway and result in dramatic cell death of recombinant Escherichia coli (Z.B. Fu, K.L. Ng, T.L. Lam, W.K.R. Wong, Cell death caused by hyper-expression of a secretory exoglucanase in Esherichia coli, Protein Expr. Purif. 42 (2005) 67-77). We propose here that the cell lysate ratio (Pre/Mat RQ) of the unprocessed precursor Exg protein (Pre-Exg) and its processed mature product (Mat-Exg) reflects the capacity of E. coli to secrete Exg. A Pre/Mat RQ of 20/80, designated the "Critical Value," was an important threshold measurement. A rise in the Pre/Mat RQ triggered a mass killing effect. The use of various secretion signal peptides did not improve the viability of cells expressing high levels of Pre-Exg under strong tac promoter control. However, use of the weaker vegG promoter in conjunction with a change in start codon of the spa leader sequence from ATG to TTG in a pM1vegGcexL plasmid construct resulted in a high level (0.9 U ml(-1)) of excreted Exg in shake-flask cultures. This was 50% higher than the best result obtained from plasmid construct lacUV5par8cex, using the lacUV5 promoter and the ompA leader sequence. Variations in the excreted Exg activities were attributable to differences in the Pre/Mat RQ values of the induced cultures harboring pM1vegGcexL and lacUV5par8cex. These values were 18/82 and 10/90, respectively. Employing fed-batch cultivation in two-liter fermentors, an induced JM101(pM1vegGcexL) culture yielded 4.5 U ml(-1) of excreted Exg, which was over six fold greater that previously reported. Our results illustrate the successful application of the Pre/Mat RQ ratio as a guide to the attainment of a maximum level of secreted/excreted Exg.

21 CFR 177.2490 - Polyphenylene sulfide resins.

Code of Federal Regulations, 2014 CFR

2014-04-01

... titled “Oxygen Flask Combustion-Gravimetric Method for Determination of Sulfur in Organic Compounds... Park, MD 20740, or available for inspection at the National Archives and Records Administration (NARA....archives.gov/federal_register/code_of_federal_regulations/ibr_locations.html. (1) Sulfur content: 28.2-29.1...

Temperature controller for crystal resonators

NASA Technical Reports Server (NTRS)

Turlington, T. R.

1980-01-01

Controller operates on less than 5W prime power and heats crystal from -10 C to 75 C in less than 45s. Unit is faster and more accurate (to within 0.7 C) than other inexpensive controllers and faster and less expensive than very precise controllers in vacuum flasks.

EFFICACY OF COMMERCIAL INOCULA IN ENHANCING BIODEGRADATION OF WEATHERED CRUDE OIL CONTAMINATING A PRINCE WILLIAM SOUND BEACH

EPA Science Inventory

In a laboratory study evaluating the effectiveness of 10 commercial products in stimulating enhanced biodegradation of Alaska North Slope crude oil, two of the products provided significantly greater alkane degradation in closed flasks than indigenous Alaskan bacterial population...

EFFICACY OF COMMERCIAL PRODUCTS IN ENHANCING OIL BIODEGRADATION IN CLOSED LABORATORY REACTORS

EPA Science Inventory

A laboratory screening protocol was designed and conducted to test the efficacy of eight commercial bacterial cultures and two non-bacterial products in enhancing the biodegradation of weathered Alaska North Slope crude oil in closed flasks. Three lines of evidence were used to ...

21 CFR 172.864 - Synthetic fatty alcohols.

Code of Federal Regulations, 2014 CFR

2014-04-01

... into a 1-liter glass-stoppered volumetric flask and adjust to volume with hexane, with mixing. n...) n-Octyl; manufactured by the hydrodimerization of 1,3-butadiene, followed by catalytic hydrogenation of the resulting dienol, and distillation to produce n-octyl alcohol with a minimum purity of 99...

21 CFR 177.2490 - Polyphenylene sulfide resins.

Code of Federal Regulations, 2010 CFR

2010-04-01

...) FOOD FOR HUMAN CONSUMPTION (CONTINUED) INDIRECT FOOD ADDITIVES: POLYMERS Substances for Use Only as... specifications as determined by methods titled “Oxygen Flask Combustion-Gravimetric Method for Determination of... thermally cured at temperatures of 700 °F and above. (d) Polyphenylene sulfide resin coatings may be used in...

21 CFR 177.2490 - Polyphenylene sulfide resins.

Code of Federal Regulations, 2012 CFR

2012-04-01

...) FOOD FOR HUMAN CONSUMPTION (CONTINUED) INDIRECT FOOD ADDITIVES: POLYMERS Substances for Use Only as... specifications as determined by methods titled “Oxygen Flask Combustion-Gravimetric Method for Determination of... thermally cured at temperatures of 700 °F and above. (d) Polyphenylene sulfide resin coatings may be used in...

21 CFR 177.2490 - Polyphenylene sulfide resins.

Code of Federal Regulations, 2011 CFR

2011-04-01

...) FOOD FOR HUMAN CONSUMPTION (CONTINUED) INDIRECT FOOD ADDITIVES: POLYMERS Substances for Use Only as... specifications as determined by methods titled “Oxygen Flask Combustion-Gravimetric Method for Determination of... thermally cured at temperatures of 700 °F and above. (d) Polyphenylene sulfide resin coatings may be used in...

21 CFR 177.2490 - Polyphenylene sulfide resins.

Code of Federal Regulations, 2013 CFR

2013-04-01

...) FOOD FOR HUMAN CONSUMPTION (CONTINUED) INDIRECT FOOD ADDITIVES: POLYMERS Substances for Use Only as... specifications as determined by methods titled “Oxygen Flask Combustion-Gravimetric Method for Determination of... thermally cured at temperatures of 700 °F and above. (d) Polyphenylene sulfide resin coatings may be used in...

78 FR 4844 - Notice of Intent To Suspend Certain Pesticide Registrations

Federal Register 2010, 2011, 2012, 2013, 2014

2013-01-23

... water. 830.7550 Partition coefficient (n- 6/16/09 6/25/09 3/16/10 1,3 octanol/water) shake flask method. 830.7570 Partition coefficient (n- 6/16/09 6/25/09 3/16/10 1,3 octanol/water) estimation by liquid...

A novel inhibitor of Lactobacillus biofilms prevents stuck fermentations in a shake flask model

USDA-ARS?s Scientific Manuscript database

Yeast ethanol fermentations contain contaminating bacteria and yeast, with Lactobacilli being a frequent contaminant. These bacteria tolerate the low pH and high ethanol concentrations present in the fermentation, and can decrease the ethanol yield. Fermentations are routinely treated with antibioti...

Production of a recombinant phospholipase A2 in Escherichia coli using resonant acoustic mixing that improves oxygen transfer in shake flasks.

PubMed

Valdez-Cruz, Norma A; Reynoso-Cereceda, Greta I; Pérez-Rodriguez, Saumel; Restrepo-Pineda, Sara; González-Santana, Jesus; Olvera, Alejandro; Zavala, Guadalupe; Alagón, Alejandro; Trujillo-Roldán, Mauricio A

2017-07-25

Shake flasks are widely used during the development of bioprocesses for recombinant proteins. Cultures of recombinant Escherichia coli with orbital mixing (OM) have an oxygen limitation negatively affecting biomass growth and recombinant-protein production. With the aim to improve mixing and aeration in shake flask cultures, we analyzed cultures subjected to OM and the novel resonant acoustic mixing (RAM) by applying acoustic energy to E. coli BL21-Gold (DE3): a producer of recombinant phospholipase A2 (rPLA2) from Micrurus laticollaris snake venom. Comparing OM with RAM (200 rpm vs. 7.5g) at the same initial volumetric oxygen transfer coefficient (k L a ≈ 80 h -1 ) ~69% less biomass was obtained with OM compared with RAM. We analyzed two more conditions increasing agitation until maximal speed (12.5 and 20g), and ~1.6- and ~1.4-fold greater biomass was obtained as compared with cultures at 7.5g. Moreover, the specific growth rate was statistically similar in all cultures carried out in RAM, but ~1.5-fold higher than that in cultures carried out under OM. Almost half of the glucose was consumed in OM, whereas between 80 and 100% of the glucose was consumed in RAM cultures, doubling biomass per glucose yields. Differential organic acid production was observed, but acetate production was prevented at the maximal RAM (20g). The amount of rPLA2 in both, OM and RAM cultures, represented 38 ± 5% of the insoluble protein. A smaller proportion of α-helices and β-sheet of purified inclusion bodies (IBs) were appreciated by ATR-FTIR from cultures carried out under OM, than those from RAM. At maximal agitation by RAM, internal E. coli localization patterns of protein aggregation changed, as well as, IBs proteolytic degradation, in conjunction with the formation of small external vesicles, although these changes did not significantly affect the cell survival response. In moderate-cell-density recombinant E. coli BL21-Gold (DE3) cultures, the agitation increases in RAM (up to the maximum) was not enough to avoid the classical oxygen limitation that happens in OM shake flasks. However, RAM presents a decrease of oxygen limitation, resulting in a favorable effect on biomass growth and volumetric rPLA2 production. While under OM a higher recombinant protein yield was obtained.

Improved ceramic slip casting technique. [application to aircraft model fabrication

NASA Technical Reports Server (NTRS)

Buck, Gregory M. (Inventor); Vasquez, Peter (Inventor)

1993-01-01

A primary concern in modern fluid dynamics research is the experimental verification of computational aerothermodynamic codes. This research requires high precision and detail in the test model employed. Ceramic materials are used for these models because of their low heat conductivity and their survivability at high temperatures. To fabricate such models, slip casting techniques were developed to provide net-form, precision casting capability for high-purity ceramic materials in aqueous solutions. In previous slip casting techniques, block, or flask molds made of plaster-of-paris were used to draw liquid from the slip material. Upon setting, parts were removed from the flask mold and cured in a kiln at high temperatures. Casting detail was usually limited with this technique -- detailed parts were frequently damaged upon separation from the flask mold, as the molded parts are extremely delicate in the uncured state, and the flask mold is inflexible. Ceramic surfaces were also marred by 'parting lines' caused by mold separation. This adversely affected the aerodynamic surface quality of the model as well. (Parting lines are invariably necessary on or near the leading edges of wings, nosetips, and fins for mold separation. These areas are also critical for flow boundary layer control.) Parting agents used in the casting process also affected surface quality. These agents eventually soaked into the mold, the model, or flaked off when releasing the case model. Different materials were tried, such as oils, paraffin, and even an algae. The algae released best, but some of it remained on the model and imparted an uneven texture and discoloration on the model surface when cured. According to the present invention, a wax pattern for a shell mold is provided, and an aqueous mixture of a calcium sulfate-bonded investment material is applied as a coating to the wax pattern. The coated wax pattern is then dried, followed by curing to vaporize the wax pattern and leave a shell mold of the calcium sulfate-bonded investment material. The shell mold is cooled to room temperature, and a ceramic slip is poured therein. After a ceramic shell of desired thickness has set up in the shell mold, excess ceramic slip is poured out. While still wet, the shell mold is peeled from the ceramic shell to expose any delicate or detailed parts, after which the ceramic shell is cured to provide a complete, detailed, precision ceramic article without parting lines.

Mount makes liquid nitrogen-cooled gamma ray detector portable

NASA Technical Reports Server (NTRS)

Fessler, T. E.

1966-01-01

Liquid nitrogen-cooled gamma ray detector system is made portable by attaching the detector to a fixture which provides a good thermal conductive path between the detector and the liquid nitrogen in a dewar flask and a low heat leak path between the detector and the external environment.

21 CFR 178.3620 - Mineral oil.

Code of Federal Regulations, 2010 CFR

2010-04-01

... extract in a 10-millimeter cell in the range from 260-350 mµ, inclusive, compared to the solvent control... hole bored in the center to closely fit the stem of the chromatographic tube. Suction flask. 250... surface of contained liquid to be evaporated. Spectrophotometric cells. Fused quartz cells, optical path...

Engineering Considerations for Hydroxide Treatment of Training Ranges

DTIC Science & Technology

2007-06-01

solutions were compared to the untreated controls. [14C] labeled samples were counted on a Packard Instruments liquid scin - tillation counter (Model...and the soil was removed to a scin - tillation vial. Unlabeled flasks had the soil and liquid analyzed for TOC and the liquid analyzed for anion content

49 CFR 173.162 - Gallium.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 49 Transportation 2 2014-10-01 2014-10-01 false Gallium. 173.162 Section 173.162 Transportation... PACKAGINGS Non-bulk Packaging for Hazardous Materials Other Than Class 1 and Class 7 § 173.162 Gallium. (a) Except when packaged in cylinders or steel flasks, gallium must be packaged in packagings which meet the...

49 CFR 173.162 - Gallium.

Code of Federal Regulations, 2012 CFR

2012-10-01

... 49 Transportation 2 2012-10-01 2012-10-01 false Gallium. 173.162 Section 173.162 Transportation... PACKAGINGS Non-bulk Packaging for Hazardous Materials Other Than Class 1 and Class 7 § 173.162 Gallium. (a) Except when packaged in cylinders or steel flasks, gallium must be packaged in packagings which meet the...

49 CFR 173.162 - Gallium.

Code of Federal Regulations, 2013 CFR

2013-10-01

... 49 Transportation 2 2013-10-01 2013-10-01 false Gallium. 173.162 Section 173.162 Transportation... PACKAGINGS Non-bulk Packaging for Hazardous Materials Other Than Class 1 and Class 7 § 173.162 Gallium. (a) Except when packaged in cylinders or steel flasks, gallium must be packaged in packagings which meet the...

Understanding Sonoluminescence

NASA Astrophysics Data System (ADS)

Brennan, Thomas

2016-12-01

Sonoluminescence is the transformation of sound into light. To most who know how to do sonoluminescence, it's just a little glowing bubble levitating in a flask of water. But it holds some surprises that have been overlooked. This book looks to reform our scientific understanding of sonoluminescence and explore the practical applications as an energy source.

High-titer production and strong antimicrobial activity of sophorolipids from Rhodotorula bogoriensis

USDA-ARS?s Scientific Manuscript database

Rhodotorula bogoriensis produces sophorolipids (SLs) that contain 13-hydroxydocosanoic acid (OH-C22) as the lipid moiety. A systematic study was conducted to further understand the fermentative production of SLs containing OH-C22 (C22-SL) by R. bogoriensis. Shake-flask studies showed that R. bogor...

21 CFR 178.3620 - Mineral oil.

Code of Federal Regulations, 2013 CFR

2013-04-01

... extract in a 10-millimeter cell in the range from 260-350 mµ, inclusive, compared to the solvent control... hole bored in the center to closely fit the stem of the chromatographic tube. Suction flask. 250... surface of contained liquid to be evaporated. Spectrophotometric cells. Fused quartz cells, optical path...

21 CFR 178.3620 - Mineral oil.

Code of Federal Regulations, 2014 CFR

2014-04-01

... cell in the range from 260-350 mµ, inclusive, compared to the solvent control. If the absorbance so... closely fit the stem of the chromatographic tube. Suction flask. 250-milliliter or 500-milliliter filter... evaporated. Spectrophotometric cells. Fused quartz cells, optical path length in the range of 5,000...

21 CFR 178.3620 - Mineral oil.

Code of Federal Regulations, 2011 CFR

2011-04-01

... extract in a 10-millimeter cell in the range from 260-350 mµ, inclusive, compared to the solvent control... hole bored in the center to closely fit the stem of the chromatographic tube. Suction flask. 250... surface of contained liquid to be evaporated. Spectrophotometric cells. Fused quartz cells, optical path...

21 CFR 178.3620 - Mineral oil.

Code of Federal Regulations, 2012 CFR

2012-04-01

... extract in a 10-millimeter cell in the range from 260-350 mµ, inclusive, compared to the solvent control... hole bored in the center to closely fit the stem of the chromatographic tube. Suction flask. 250... surface of contained liquid to be evaporated. Spectrophotometric cells. Fused quartz cells, optical path...

77 FR 46289 - Technical Corrections to Organizational Names, Addresses, and OMB Control Numbers

Federal Register 2010, 2011, 2012, 2013, 2014

2012-08-03

...]795.232 Inhalation and dermal pharmacokinetics of commercial hexane. * * * * * (c) * * * (2) * * * (i... to read as follows: Sec. 799.6755 TSCA partition coefficient (n-octanol/water), shake flask method... read as follows: Sec. 799.6756 TSCA partition coefficient (n-octanol/water), generator column method...

AN EQUATORIAL PACIFIC RAIN EVENT: INFLUENCE ON THE DISTRIBUTION OF IRON AND HYDROGEN PEROXIDE IN SURFACE WATERS

EPA Science Inventory

Methods are described for measuring changes in atmospheric O2 concentration with emphasis on gas handling procedures. Cryogenically dried air samples are collected in 5 L glass flasks at ambient pressure and analyzed against reference gases derived from high-pressure aluminum tan...

40 CFR 796.3100 - Aerobic aquatic biodegradation.

Code of Federal Regulations, 2014 CFR

2014-07-01

... Aerobic aquatic biodegradation. (a) Introduction—(1) Purpose. (i) This Guideline is designed to develop... biodegradability of a series of functionally or structurally related chemicals, media from all inoculum flasks may..., and control system should be analyzed at time zero and at a minimum of four other times from time zero...

40 CFR 796.3100 - Aerobic aquatic biodegradation.

Code of Federal Regulations, 2012 CFR

2012-07-01

... Aerobic aquatic biodegradation. (a) Introduction—(1) Purpose. (i) This Guideline is designed to develop... biodegradability of a series of functionally or structurally related chemicals, media from all inoculum flasks may..., and control system should be analyzed at time zero and at a minimum of four other times from time zero...

40 CFR 796.3100 - Aerobic aquatic biodegradation.

Code of Federal Regulations, 2013 CFR

2013-07-01

... Aerobic aquatic biodegradation. (a) Introduction—(1) Purpose. (i) This Guideline is designed to develop... biodegradability of a series of functionally or structurally related chemicals, media from all inoculum flasks may..., and control system should be analyzed at time zero and at a minimum of four other times from time zero...

Spider Silk Glue Proteins BAA 8.1

DTIC Science & Technology

2017-09-14

protein expression. 3. The genes were then used in flask fermentation expression studies to insure that protein of the correct size was being...one of the lengths (3X, roughly a 140kD protein) for initial studies. We were able to detect protein production in these fermentations but we saw

OIL SPILL DISPERSANT EFFECTIVENESS PROTOCOL. II: PERFORMANCE OF THE REVISED PROTOCOL

EPA Science Inventory

The current U.S. Environmental Protection Agency (EPA) protocol for testing the effectiveness of dispersants for use in treating oil spills on the open water, the swirling flask test (SFT), has been found to give widely varying results in the hands of different testing laborator...

A Kinetic Study of the Adiabatic Polymerization of Acrylamide.

ERIC Educational Resources Information Center

Thomson, R. A. M.

1986-01-01

Discusses theory, procedures, and results for an experiment which demonstrates the application of basic physics to chemical problems. The experiment involves the adiabatic process, in which polymerization carried out in a vacuum flask is compared to the theoretical prediction of the model with the temperature-time curve obtained in practice. (JN)

40 CFR 799.6755 - TSCA partition coefficient (n-octanol/water), shake flask method.

Code of Federal Regulations, 2013 CFR

2013-07-01

...) Qualifying statements. This method applies only to pure, water soluble substances which do not dissociate or... applies to a pure substance dispersed between two pure solvents. If several different solutes occur in one... applied. The values presented in table 1 of this section are not necessarily representative of the results...

40 CFR 799.6755 - TSCA partition coefficient (n-octanol/water), shake flask method.

Code of Federal Regulations, 2014 CFR

2014-07-01

...) Qualifying statements. This method applies only to pure, water soluble substances which do not dissociate or... applied. The values presented in table 1 of this section are not necessarily representative of the results... Law applies only at constant temperature, pressure, and pH for dilute solutions. It strictly applies...

Conversion of canola meal into a high-protein feed additive via solid-state fungal incubation process

USDA-ARS?s Scientific Manuscript database

The study goal was to determine the optimal fungal culture to reduce glucosinolates (GLS), fiber, and residual sugars while increasing the protein content and nutritional value of canola meal. Solid-state incubation conditions were used to enhance filamentous growth of the fungi. Flask trials were p...

USE OF THE MIXED FLASK CULTURE (MFC) MICROCOSM PROTOCOL TO ESTIMATE THE SURVIVAL AND EFFECTS OF MICROORGANISMS ADDED TO FRESHWATER ECOSYSTEMS

EPA Science Inventory

The ability to manipulate an organism's genetic substance offers benefits to many aspects of human health and well-being. oupled with this positive aspect of genetic engineering, however, is a concern about potential adverse effects on human welfare and environmental quality. ive...

Glycerine and levulinic acid: renewable co-substrates for the fermentative synthesis of short-chain poly(hydroxyalkanoate) biopolymers

USDA-ARS?s Scientific Manuscript database

Glycerine and levulinic acid were used alone and in combination for the fermentative synthesis of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHB/V) biopolymers. Shake-flask cultures of Pseudomonas oleovorans NRRL B-14682 containing different glycerine:levulinic acid ratios (1%, w/v total carbon ...

Bond Potential of Lithium Disilicate to Heat-Cured Polymethylmethacrylate (PMMA)

DTIC Science & Technology

2015-06-01

system eliminates the need for flasking, boil-out, and long processing times. Instead, the base is light-cured while the teeth are placed with a VLC set...component of previous studies causing failure can be challenged . The stronger ceramic may be durable enough to more accurately measure the bond strength

Field Portable Methods for Rapid Water Quality Analysis

DTIC Science & Technology

1996-10-01

a pie-shaped segment of the rolled cylinder, also stored in a sterile flask. Several membrane layers were pulled away, and a scraping was removed for...For example, the H. pylori urease A gene can be used, and thefla A gene can be used for the campylobacter species. 12 2. Implement microwave technology

21 CFR 172.864 - Synthetic fatty alcohols.

Code of Federal Regulations, 2012 CFR

2012-04-01

... into a 1-liter glass-stoppered volumetric flask and adjust to volume with hexane, with mixing. n... specifications prescribed in paragraph (a)(1) (i) and (iii) of this section. (3) n-Octyl; manufactured by the... produce n-octyl alcohol with a minimum purity of 99 percent. The analytical method for n-octyl alcohol...

21 CFR 172.864 - Synthetic fatty alcohols.

Code of Federal Regulations, 2011 CFR

2011-04-01

... into a 1-liter glass-stoppered volumetric flask and adjust to volume with hexane, with mixing. n... specifications prescribed in paragraph (a)(1) (i) and (iii) of this section. (3) n-Octyl; manufactured by the... produce n-octyl alcohol with a minimum purity of 99 percent. The analytical method for n-octyl alcohol...

21 CFR 172.864 - Synthetic fatty alcohols.

Code of Federal Regulations, 2013 CFR

2013-04-01

... into a 1-liter glass-stoppered volumetric flask and adjust to volume with hexane, with mixing. n... specifications prescribed in paragraph (a)(1) (i) and (iii) of this section. (3) n-Octyl; manufactured by the... produce n-octyl alcohol with a minimum purity of 99 percent. The analytical method for n-octyl alcohol...

40 CFR 799.6784 - TSCA water solubility: Column elution method; shake flask method.

Code of Federal Regulations, 2011 CFR

2011-07-01

....053 (3) The column should be connected to a recycling pump capable of controlling flows of... the carrier is not achieved due to partition effects on the surface of the carrier. (2) The loading of... this, the recycling pump is connected and the apparatus allowed to run until equilibration is...

40 CFR 799.6784 - TSCA water solubility: Column elution method; shake flask method.

Code of Federal Regulations, 2013 CFR

2013-07-01

....053 (3) The column should be connected to a recycling pump capable of controlling flows of... the carrier is not achieved due to partition effects on the surface of the carrier. (2) The loading of... this, the recycling pump is connected and the apparatus allowed to run until equilibration is...

40 CFR 799.6784 - TSCA water solubility: Column elution method; shake flask method.

Code of Federal Regulations, 2012 CFR

2012-07-01

....053 (3) The column should be connected to a recycling pump capable of controlling flows of... the carrier is not achieved due to partition effects on the surface of the carrier. (2) The loading of... this, the recycling pump is connected and the apparatus allowed to run until equilibration is...

40 CFR 799.6784 - TSCA water solubility: Column elution method; shake flask method.

Code of Federal Regulations, 2014 CFR

2014-07-01

....053 (3) The column should be connected to a recycling pump capable of controlling flows of... the carrier is not achieved due to partition effects on the surface of the carrier. (2) The loading of... this, the recycling pump is connected and the apparatus allowed to run until equilibration is...