Rat sperm motility analysis: methodologic considerations

EPA Science Inventory

The objective of these studies was to optimize conditions for computer-assisted sperm analysis (CASA) of rat epididymal spermatozoa. Methodologic issues addressed include sample collection technique, sampling region within the epididymis, type of diluent medium used, and sample c...

Role of medullary progenitor cells in epithelial cell migration and proliferation

PubMed Central

Chen, Dong; Chen, Zhiyong; Zhang, Yuning; Park, Chanyoung; Al-Omari, Ahmed

2014-01-01

This study is aimed at characterizing medullary interstitial progenitor cells and to examine their capacity to induce tubular epithelial cell migration and proliferation. We have isolated a progenitor cell side population from a primary medullary interstitial cell line. We show that the medullary progenitor cells (MPCs) express CD24, CD44, CXCR7, CXCR4, nestin, and PAX7. MPCs are CD34 negative, which indicates that they are not bone marrow-derived stem cells. MPCs survive >50 passages, and when grown in epithelial differentiation medium develop phenotypic characteristics of epithelial cells. Inner medulla collecting duct (IMCD3) cells treated with conditioned medium from MPCs show significantly accelerated cell proliferation and migration. Conditioned medium from PGE2-treated MPCs induce tubule formation in IMCD3 cells grown in 3D Matrigel. Moreover, most of the MPCs express the pericyte marker PDGFR-b. Our study shows that the medullary interstitium harbors a side population of progenitor cells that can differentiate to epithelial cells and can stimulate tubular epithelial cell migration and proliferation. The findings of this study suggest that medullary pericyte/progenitor cells may play a critical role in collecting duct cell injury repair. PMID:24808539

Long-term viability and differentiation of bovine oviductal monolayers: bidimensional versus three-dimensional culture.

PubMed

Gualtieri, R; Mollo, V; Braun, S; Barbato, V; Fiorentino, I; Talevi, R

2012-10-15

Different in vitro models have been developed to study the interaction of gametes and embryos with the maternal tract. In cattle, the interaction of the oviduct with gametes and embryos have been classically studied using oviductal explants or monolayers (OMs). Explants are well differentiated but have to be used within 24 h after collection, whereas OMs can be used for a longer time after cell confluence but dedifferentiate during culture, losing cell polarity and ciliation. Herein, OMs were cultured either in M199 plus 10% fetal calf serum or in a semidefined culture medium (Gray's medium), in an immersed condition on collagen-coated coated microporous polyester or polycarbonate inserts under air-liquid interface conditions. The influence of culture conditions on long-term viability and differentiation of OMs was evaluated through scanning electron microscopy, localization of centrin and tubulin at the confocal laser scanning microscope, and assessment of maintenance of viability of sperm bound to OMs. Findings demonstrated that OMs cultured in an immersed condition with Gray's medium retain a better morphology, do not exhibit signs of crisis at least until 3 wks postconfluence, and maintain the viability of bound sperm significantly better than parallel OMs cultured in M199 plus 10% fetal calf serum. OM culture with Gray's medium in air-liquid interface conditions on porous inserts promotes cell polarity, ciliation, and maintenance of bound sperm viability at least until 3 wks postconfluence. In conclusion, oviduct culture in Gray's medium in an immersed or air-liquid condition allows long-term culture and, in the latter case, also ciliation of bovine OMs, and may represent in vitro systems that mimick more closely the biological processes modulated by the oviduct in vivo. Copyright © 2012 Elsevier Inc. All rights reserved.

Method, system, and computer-readable medium for determining performance characteristics of an object undergoing one or more arbitrary aging conditions

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

Gering, Kevin L.

A method, system, and computer-readable medium are described for characterizing performance loss of an object undergoing an arbitrary aging condition. Baseline aging data may be collected from the object for at least one known baseline aging condition over time, determining baseline multiple sigmoid model parameters from the baseline data, and performance loss of the object may be determined over time through multiple sigmoid model parameters associated with the object undergoing the arbitrary aging condition using a differential deviation-from-baseline approach from the baseline multiple sigmoid model parameters. The system may include an object, monitoring hardware configured to sample performance characteristics ofmore » the object, and a processor coupled to the monitoring hardware. The processor is configured to determine performance loss for the arbitrary aging condition from a comparison of the performance characteristics of the object deviating from baseline performance characteristics associated with a baseline aging condition.« less

Occupational Safety and Health Conditions Aboard Small- and Medium-Size Fishing Vessels: Differences among Age Groups.

PubMed

Zytoon, Mohamed A; Basahel, Abdulrahman M

2017-02-24

Although marine fishing is one of the most hazardous occupations, research on the occupational safety and health (OSH) conditions aboard marine fishing vessels is scarce. For instance, little is known about the working conditions of vulnerable groups such as young and aging fishermen. The objective of the current paper is to study the OSH conditions of young and aging fishermen compared to middle-aged fishermen in the small- and medium-size (SM) marine fishing sector. A cross-sectional study was designed, and 686 fishermen working aboard SM fishing vessels were interviewed to collect information about their safety and health. The associations of physical and psychosocial work conditions with safety and health outcomes, e.g., injuries, illnesses and job satisfaction, are presented. The results of the current study can be utilized in the design of effective accident prevention and OSH training programs for the three age groups and in the regulation of working conditions aboard fishing vessels.

Occupational Safety and Health Conditions Aboard Small- and Medium-Size Fishing Vessels: Differences among Age Groups

PubMed Central

Zytoon, Mohamed A.; Basahel, Abdulrahman M.

2017-01-01

Although marine fishing is one of the most hazardous occupations, research on the occupational safety and health (OSH) conditions aboard marine fishing vessels is scarce. For instance, little is known about the working conditions of vulnerable groups such as young and aging fishermen. The objective of the current paper is to study the OSH conditions of young and aging fishermen compared to middle-aged fishermen in the small- and medium-size (SM) marine fishing sector. A cross-sectional study was designed, and 686 fishermen working aboard SM fishing vessels were interviewed to collect information about their safety and health. The associations of physical and psychosocial work conditions with safety and health outcomes, e.g., injuries, illnesses and job satisfaction, are presented. The results of the current study can be utilized in the design of effective accident prevention and OSH training programs for the three age groups and in the regulation of working conditions aboard fishing vessels. PMID:28245578

77 FR 75257 - Proposed Collection of Information: Medium- and Heavy-Duty Truck Fleet Survey

Federal Register 2010, 2011, 2012, 2013, 2014

2012-12-19

...-0170] Proposed Collection of Information: Medium- and Heavy-Duty Truck Fleet Survey AGENCY: National... collection of information will be in the form of a one-time survey of medium- and heavy-duty truck fleet... collection. OMB Control Number: To be issued at time of approval. Title: Medium- and Heavy-Duty Truck Fleet...

Bacillus licheniformis BT5.9 Isolated from Changar Hot Spring, Malang, Indonesia, as a Potential Producer of Thermostable α-amylase

PubMed Central

Ibrahim, Darah; Zhu, Han Li; Yusof, Nuraqilah; Isnaeni; Hong, Lim Sheh

2013-01-01

A total of 34 bacterial isolates were obtained from soil samples collected from Changar Hot Spring, Malang, Indonesia. Of these, 13 isolates produced a zone of hydrolysis in starch-nutrient agar medium and generated various amylases in liquid medium. One isolate was selected as the best amylase producer and was identified as Bacillus licheniformis BT5.9. The improvement of culture conditions (initial medium pH of 5.0, cultivation temperature of 50°C, agitation speed of 100 rpm and inoculum size of 1.7 × 109 cells/ml) provided the highest amylase production (0.327 U/ml). PMID:24575243

Lung cells support osteosarcoma cell migration and survival.

PubMed

Yu, Shibing; Fourman, Mitchell Stephen; Mahjoub, Adel; Mandell, Jonathan Brendan; Crasto, Jared Anthony; Greco, Nicholas Giuseppe; Weiss, Kurt Richard

2017-01-25

Osteosarcoma (OS) is the most common primary bone tumor, with a propensity to metastasize to the lungs. Five-year survival for metastatic OS is below 30%, and has not improved for several decades despite the introduction of multi-agent chemotherapy. Understanding OS cell migration to the lungs requires an evaluation of the lung microenvironment. Here we utilized an in vitro lung cell and OS cell co-culture model to explore the interactions between OS and lung cells, hypothesizing that lung cells would promote OS cell migration and survival. The impact of a novel anti-OS chemotherapy on OS migration and survival in the lung microenvironment was also examined. Three human OS cell lines (SJSA-1, Saos-2, U-2) and two human lung cell lines (HULEC-5a, MRC-5) were cultured according to American Type Culture Collection recommendations. Human lung cell lines were cultured in growth medium for 72 h to create conditioned media. OS proliferation was evaluated in lung co-culture and conditioned media microenvironment, with a murine fibroblast cell line (NIH-3 T3) in fresh growth medium as controls. Migration and invasion were measured using a real-time cell analysis system. Real-time PCR was utilized to probe for Aldehyde Dehydrogenase (ALDH1) expression. Osteosarcoma cells were also transduced with a lentivirus encoding for GFP to permit morphologic analysis with fluorescence microscopy. The anti-OS efficacy of Disulfiram, an ALDH-inhibitor previously shown to inhibit OS cell proliferation and metastasis in vitro, was evaluated in each microenvironment. Lung-cell conditioned medium promoted osteosarcoma cell migration, with a significantly higher attractive effect on all three osteosarcoma cell lines compared to basic growth medium, 10% serum containing medium, and NIH-3 T3 conditioned medium (p <0.05). Lung cell conditioned medium induced cell morphologic changes, as demonstrated with GFP-labeled cells. OS cells cultured in lung cell conditioned medium had increased alkaline phosphatase staining. Lung endothelial HULEC-5a cells are attractants for OS cell migration, proliferation, and survival. The SJSA-1 osteosarcoma cell line demonstrated greater metastatic potential than Saos-2 and U-2 cells. ALDH appears to be involved in the interaction between lung and OS cells, and ALP may be a valuable biomarker for monitoring functional OS changes during metastasis.

Medium-term in vitro storage as a complementary germplasm preservation technique

USDA-ARS?s Scientific Manuscript database

A Germplasm preservation of vegetatively propagated crops may be accomplished using a range of old and new technologies. Field collections, potted plants, or some cases whole plants stored under cool to cold conditions are complemented by in vitro culture, in vitro storage, and cryopreservation. The...

Endotoxin-induced lung alveolar cell injury causes brain cell damage.

PubMed

Rodríguez-González, Raquel; Ramos-Nuez, Ángela; Martín-Barrasa, José Luis; López-Aguilar, Josefina; Baluja, Aurora; Álvarez, Julián; Rocco, Patricia R M; Pelosi, Paolo; Villar, Jesús

2015-01-01

Sepsis is the most common cause of acute respiratory distress syndrome, a severe lung inflammatory disorder with an elevated morbidity and mortality. Sepsis and acute respiratory distress syndrome involve the release of inflammatory mediators to the systemic circulation, propagating the cellular and molecular response and affecting distal organs, including the brain. Since it has been reported that sepsis and acute respiratory distress syndrome contribute to brain dysfunction, we investigated the brain-lung crosstalk using a combined experimental in vitro airway epithelial and brain cell injury model. Conditioned medium collected from an in vitro lipopolysaccharide-induced airway epithelial cell injury model using human A549 alveolar cells was subsequently added at increasing concentrations (no conditioned, 2%, 5%, 10%, 15%, 25%, and 50%) to a rat mixed brain cell culture containing both astrocytes and neurons. Samples from culture media and cells from mixed brain cultures were collected before treatment, and at 6 and 24 h for analysis. Conditioned medium at 15% significantly increased apoptosis in brain cell cultures 24 h after treatment, whereas 25% and 50% significantly increased both necrosis and apoptosis. Levels of brain damage markers S100 calcium binding protein B and neuron-specific enolase, interleukin-6, macrophage inflammatory protein-2, as well as matrix metalloproteinase-9 increased significantly after treating brain cells with ≥2% conditioned medium. Our findings demonstrated that human epithelial pulmonary cells stimulated with bacterial lipopolysaccharide release inflammatory mediators that are able to induce a translational clinically relevant and harmful response in brain cells. These results support a brain-lung crosstalk during sepsis and sepsis-induced acute respiratory distress syndrome. © 2014 by the Society for Experimental Biology and Medicine.

Sensitive and selective culture medium for detection of environmental Clostridium difficile isolates without requirement for anaerobic culture conditions.

PubMed

Cadnum, Jennifer L; Hurless, Kelly N; Deshpande, Abhishek; Nerandzic, Michelle M; Kundrapu, Sirisha; Donskey, Curtis J

2014-09-01

Effective and easy-to-use methods for detecting Clostridium difficile spore contamination would be useful for identifying environmental reservoirs and monitoring the effectiveness of room disinfection. Culture-based detection methods are sensitive for detecting C. difficile, but their utility is limited due to the requirement of anaerobic culture conditions and microbiological expertise. We developed a low-cost selective broth medium containing thioglycolic acid and l-cystine, termed C. difficile brucella broth with thioglycolic acid and l-cystine (CDBB-TC), for the detection of C. difficile from environmental specimens under aerobic culture conditions. The sensitivity and specificity of CDBB-TC (under aerobic culture conditions) were compared to those of CDBB (under anaerobic culture conditions) for the recovery of C. difficile from swabs collected from hospital room surfaces. CDBB-TC was significantly more sensitive than CDBB for recovering environmental C. difficile (36/41 [88%] versus 21/41 [51%], respectively; P = 0.006). C. difficile latex agglutination, an enzyme immunoassay for toxins A and B or glutamate dehydrogenase, and a PCR for toxin B genes were all effective as confirmatory tests. For 477 total environmental cultures, the specificity of CDBB-TC versus that of CDBB based upon false-positive yellow-color development of the medium without recovery of C. difficile was 100% (0 false-positive results) versus 96% (18 false-positive results), respectively. False-positive cultures for CDBB were attributable to the growth of anaerobic non-C. difficile organisms that did not grow in CDBB-TC. Our results suggest that CDBB-TC provides a sensitive and selective medium for the recovery of C. difficile organisms from environmental samples, without the need for anaerobic culture conditions. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Collection and preservation of pygmy hippopotamus (Choeropsis liberiensis) semen.

PubMed

Saragusty, J; Hildebrandt, T B; Bouts, T; Göritz, F; Hermes, R

2010-09-01

Knowledge about the reproduction of the endangered pygmy hippopotamus is almost non-existent. This study takes the first step toward changing this by devising a protocol for the collection, evaluation, and short-term preservation of semen of this endangered species. Semen was collected successfully from seven bulls by electroejaculation, using a specially designed rectal probe. Mean +/- SEM values of native sperm parameters from combined best fractions were: motility-80.0 +/- 4.1%, concentration-2421 +/- 1530 x 10(6) cells/mL, total collected cell number-759 +/- 261 x 10(6) cells, intact acrosome-87.8 +/- 1.2%, intact morphology-52.7 +/- 4.3%, and, for some, hypoosmotic swelling test-79.3 +/- 4.4% and seminal plasma osmolarity-297.5 +/- 3.3 mOsm. Seven different extenders were tested for sperm storage under chilling conditions: Berliner Cryomedium (BC), Biladyl, modification of Kenney modified Tyrode's medium (KMT), MES medium, Androhep((R)), boar M III() extender and Human Sperm Refrigeration Medium. While differences between males were apparent, the BC was consistently superior to all other extenders in sperm motility and facilitated storage for 7 d with up to 30% motility and some motility even after 3 weeks. With this knowledge in hand, the obvious two directions for future research are to conduct artificial insemination and to develop a technique for sperm cryopreservation. Copyright 2010 Elsevier Inc. All rights reserved.

Medium chain triglycerides dose-dependently prevent liver pathology in a rat model of nonalcoholic fatty liver disease

USDA-ARS?s Scientific Manuscript database

Obesity is often associated with a cluster of increased health risks collectively known as "Metabolic Syndrome" (MS). MS is often accompanied by development of fatty liver. Sometimes fatty liver results in damage leading to reduced liver function, and need for a transplant. This condition is known...

Method for collecting spores from a mold

DOEpatents

Au, Frederick H. F.; Beckert, Werner F.

1977-01-01

A technique and apparatus used therewith for determining the uptake of plutonium and other contaminants by soil microorganisms which, in turn, gives a measure of the plutonium and/or other contaminants available to the biosphere at that particular time. A measured quantity of uncontaminated spores of a selected mold is added to a moistened sample of the soil to be tested. The mixture is allowed to sit a predetermined number of days under specified temperature conditions. An agar layer is then applied to the top of the sample. After three or more days, when spores of the mold growing in the sample have formed, the spores are collected by a miniature vacuum collection apparatus operated under preselected vacuum conditions, which collect only the spores with essentially no contamination by mycelial fragments or culture medium. After collection, the fungal spores are dried and analyzed for the plutonium and/or other contaminants. The apparatus is also suitable for collection of pollen, small insects, dust and other small particles, material from thin-layer chromatography plates, etc.

Testicular cell conditioned medium supports differentiation of embryonic stem cells into ovarian structures containing oocytes.

PubMed

Lacham-Kaplan, Orly; Chy, Hun; Trounson, Alan

2006-02-01

Previous reports and the current study have found that germ cell precursor cells appear in embryoid bodies (EBs) formed from mouse embryonic stem cells as identified by positive expression of specific germ cell markers such as Oct-3/4, Mvh, c-kit, Stella, and DAZL. We hypothesized that if exposed to appropriate growth factors, the germ cell precursor cells within the EBs would differentiate into gametes. The source for growth factors used in the present study is conditioned medium collected from testicular cell cultures prepared from the testes of newborn males. Testes at this stage of development contain most growth factors required for the transformation of germ stem cells into differentiated gametes. When EBs were cultured in the conditioned medium, they developed into ovarian structures, which contained putative oocytes. The oocytes were surrounded by one to two layers of flattened cells and did not have a visible zona pellucida. However, oocyte-specific markers such as Fig-alpha and ZP3 were found expressed by the ovarian structures. The production of oocytes using this method is repeatable and reliable and may be applicable to other mammalian species, including the human.

Triolein reduces MMP-1 upregulation in dermal fibroblasts generated by ROS production in UVB-irradiated keratinocytes.

PubMed

Leirós, Gustavo J; Kusinsky, Ana Gabriela; Balañá, María Eugenia; Hagelin, Karin

2017-02-01

Cytokine production and oxidative stress generated by ultraviolet radiation B (UVB) skin exposure are main factors of skin photoaging. Interleukin-6 (IL-6) produced by irradiated keratinocytes is proposed to have a role in metalloproteinases (MMPs) expression activation in dermal fibroblasts. We examined the effect of triolein treatment of UVB-irradiated keratinocytes on MMP1 (interstitial collagenase) expression response of dermal fibroblasts. We assayed UVB-irradiated keratinocytes soluble signals, mainly IL-6 and reactive oxygen species (ROS). IL-6 expression and ROS generation were assayed in UVB-irradiated keratinocytes. MMP1 mRNA expression response was assayed in fibroblasts grown in keratinocytes conditioned medium. We evaluated the effect of treating keratinocytes with triolein on IL-6 expression and ROS generation in keratinocytes, and MMP1 expression in fibroblasts. The irradiation of epidermal cells with sublethal UVB doses increased IL-6 expression and ROS generation. Conditioned culture medium collected from keratinocytes was used to culture dermal fibroblasts. MMP1 mRNA expression increase was observed in fibroblasts cultured in medium collected from UVB-irradiated keratinocytes. Triolein treatment reduced the IL-6 expression and ROS generation in keratinocytes and this effect was reflected in downregulation of MMP1 expression in fibroblasts. Triolein reduces both the expression of IL-6 and ROS generation in irradiated keratinocytes. It seems to exert an anti-inflammatory and anti-oxidative stress effect on irradiated keratinocytes that in turn reduces MMP1 expression in dermal fibroblasts. Collectively, these results indicate that triolein could act as a photoprotective agent. Copyright © 2016 Japanese Society for Investigative Dermatology. Published by Elsevier B.V. All rights reserved.

Enhancing Lexical Knowledge through L2 Medium Tasks

ERIC Educational Resources Information Center

Vosoughi, Marjan

2012-01-01

The purpose of the present study was to investigate the L2 use rate in EFL classrooms through introducing three task conditions in learning lexical items. Data were collected from a group of freshman university students (male and female) studying in Islamic Azad university of Sabzevar, Iran (N = 73). Based on their performance on a Michigan TOEFL…

ELT Teacher Education Flipped Classroom: An Analysis of Task Challenge and Student Teachers' Views and Expectations

ERIC Educational Resources Information Center

Karaaslan, Hatice; Çelebi, Hatice

2017-01-01

In this study, we explore the interplay between task complexity, task conditions and task difficulty introduced by Robinson (2001) in flipped classroom instruction at tertiary level through the data we collected from undergraduate English Language Teaching (ELT) department students studying at an English-medium state university. For the…

System for sampling and monitoring microscopic organisms and substances

DOEpatents

Au, Frederick H. F.; Beckert, Werner F.

1976-01-01

A technique and apparatus used therewith for determining the uptake of plutonium and other contaminants by soil microorganisms which, in turn, gives a measure of the plutonium and/or other contaminants available to the biosphere at that particular time. A measured quantity of uncontaminated spores of a selected mold is added to a moistened sample of the soil to be tested. The mixture is allowed to sit a predetermined number of days under specified temperature conditions. An agar layer is then applied to the top of the sample. After three or more days, when spores of the mold growing in the sample have formed, the spores are collected by a miniature vacuum collection apparatus operated under preselected vacuum conditions, which collect only the spores with essentially no contamination by mycelial fragments or culture medium. After collection, the fungal spores are dried and analyzed for the plutonium and/or other contaminants. The apparatus is also suitable for collection of pollen, small insects, dust and other small particles, material from thin-layer chromatography plates, etc.

In Vitro Impact of Conditioned Medium From Demineralized Freeze-Dried Bone on Human Umbilical Endothelial Cells.

PubMed

Harnik, Branko; Miron, Richard J; Buser, Daniel; Gruber, Reinhard

2017-03-01

Angiogenesis is essential for the consolidation of bone allografts. The underlying molecular mechanism, however, remains unclear. Soluble factors released from demineralized freeze-dried bone target mesenchymal cells; however, their effect on endothelial cells has not been investigated so far. The aim of the present study was therefore to examine the effect of conditioned medium from demineralized freeze-dried bone on human umbilical endothelial cells in vitro. Conditioned medium was first prepared from demineralized freeze-dried bone following 24 hours incubation at room temperature to produce demineralized bone conditioned media. Thereafter, conditioned medium was used to stimulate human umbilical vein endothelial cells in vitro by determining the cell response based on viability, proliferation, expression of apoptotic genes, a Boyden chamber to determine cell migration, and the formation of branches. The authors report here that conditioned medium decreased viability and proliferation of endothelial cells. Neither of the apoptotic marker genes was significantly altered when endothelial cells were exposed to conditioned medium. The Boyden chamber revealed that endothelial cells migrate toward conditioned medium. Moreover, conditioned medium moderately stimulated the formation of branches. These findings support the concept that conditioned medium from demineralized freeze-dried bone targets endothelial cells by decreasing their proliferation and enhancing their motility under these in vitro conditions.

Hydrogen production using hydrogenase-containing oxygenic photosynthetic organisms

DOEpatents

Melis, Anastasios; Zhang, Liping; Benemann, John R.; Forestier, Marc; Ghirardi, Maria; Seibert, Michael

2006-01-24

A reversible physiological process provides for the temporal separation of oxygen evolution and hydrogen production in a microorganism, which includes the steps of growing a culture of the microorganism in medium under illuminated conditions to accumulate an endogenous substrate, depleting from the medium a nutrient selected from the group consisting of sulfur, iron, and/or manganese, sealing the culture from atmospheric oxygen, incubating the culture in light whereby a rate of light-induced oxygen production is equal to or less than a rate of respiration, and collecting an evolved gas. The process is particularly useful to accomplish a sustained photobiological hydrogen gas production in cultures of microorganisms, such as Chlamydomonas reinhardtii.

Hydrogen Production Using Hydrogenase-Containing Oxygenic Photosynthetic Organisms

DOEpatents

Melis, A.; Zhang, L.; Benemann, J. R.; Forestier, M.; Ghirardi, M.; Seibert, M.

2006-01-24

A reversible physiological process provides for the temporal separation of oxygen evolution and hydrogen production in a microorganism, which includes the steps of growing a culture of the microorganism in medium under illuminated conditions to accumulate an endogenous substrate, depleting from the medium a nutrient selected from the group consisting of sulfur, iron, and/or manganese, sealing the culture from atmospheric oxygen, incubating the culture in light whereby a rate of light-induced oxygen production is equal to or less than a rate of respiration, and collecting an evolved gas. The process is particularly useful to accomplish a sustained photobiological hydrogen gas production in cultures of microorganisms, such as Chlamydomonas reinhardtii.

A differential medium for the enumeration of the spoilage yeast Zygosaccharomyces bailii in wine.

PubMed

Schuller, D; Côrte-Real, M; Leão, C

2000-11-01

A collection of yeasts, isolated mostly from spoiled wines, was used in order to develop a differential medium for Zygosaccharomyces bailii. The 118 selected strains of 21 species differed in their origin and resistance to preservatives and belonged to the genera Pichia, Torulaspora, Dekkera, Debaryomyces, Saccharomycodes, Issatchenkia, Kluyveromyces, Kloeckera, Lodderomyces, Schizosaccharomyces, Rhodotorula, Saccharomyces, and Zygosaccharomyces. The design of the culture medium was based on the different ability of the various yeast species to grow in a mineral medium with glucose and formic acid (mixed-substrate medium) as the only carbon and energy sources and supplemented with an acid-base indicator. By manipulating the concentration of the acid and the sugar it was possible to select conditions where only Z. bailii strains gave rise to alkalinization, associated with a color change of the medium (positive response). The final composition of the mixed medium was adjusted as a compromise between the percentage of recovery and selectivity for Z. bailii. This was accomplished by the use of pure or mixed cultures of the yeast strains and applying the membrane filtration methodology. The microbiological analysis of two samples of contaminated Vinho Verde showed that the new medium can be considered as a differential medium to distinguish Z. bailii from other contaminating yeasts, having potential application in the microbiological control of wines and probably other beverages and foods.

An efficient in vitro regeneration protocol for a natural dye yielding plant, Strobilanthes flaccidifolious Nees., from nodal explants.

PubMed

Deb, Chitta Ranjan; Arenmongla, T

2012-11-01

Adventitious shoot buds formation from axillary buds of nodal segments of S. flaccidifolious was achieved on MS medium containing sucrose (3%, w/v), and a-naphthalene acetic acid (NAA; 3 microM) and benzyl adenine (3 microM) in combination. The nodal segments were primed on 'Growtak Sieve' for 48 h on MS medium containing sucrose (2%), polyvinyl pyrollidone (200 mgL(-1)) as antioxidant. About 80% of primed nodal segments responded positively and formed approximately 12 adventitious shoot buds per explants from explants collected during October-November months of every year. The shoot buds converted into plantlets on MS medium containing sucrose (3%) and kinetin (3 microM) where approximately 7 micro shoots developed per subculture after 8 weeks of culture. The regenerated micro shoots induced average 14 roots/plant on medium containing NAA (3 microM). The regenerates were hardened for 6-7 weeks on medium with 1/2MS salt solution and sucrose (2%) under normal laboratory condition before transferring to potting mix. About 70% transplants survived after two months of transfer.

Quantitative Characterization of the Growth of Deinococcus geothermalis DSM-11302: Effect of Inoculum Size, Growth Medium and Culture Conditions.

PubMed

Bornot, Julie; Molina-Jouve, Carole; Uribelarrea, Jean-Louis; Gorret, Nathalie

2015-08-20

Due to their remarkable resistance to extreme conditions, Deinococcaceae strains are of great interest to biotechnological prospects. However, the physiology of the extremophile strain Deinococcus geothermalis has scarcely been studied and is not well understood. The physiological behaviour was then studied in well-controlled conditions in flask and bioreactor cultures. The growth of D. geothermalis type strains was compared. Among the strains tested, the strain from the German Collection of Microorganisms (Deutsche Sammlung von Mikroorganismen DSM) DSM-11302 was found to give the highest biomass concentration and growth rate: in a complex medium with glucose, the growth rate reached 0.75 h(-1) at 45 °C. Yeast extract concentration in the medium had significant constitutive and catalytic effects. Furthermore, the results showed that the physiological descriptors were not affected by the inoculum preparation steps. A batch culture of D. geothermalis DSM-11302 on defined medium was carried out: cells grew exponentially with a maximal growth rate of 0.28 h(-1) and D. geothermalis DSM-11302 biomass reached 1.4 g·L(-1) in 20 h. Then, 1.4 gDryCellWeight of biomass (X) was obtained from 5.6 g glucose (Glc) consumed as carbon source, corresponding to a yield of 0.3 CmolX·CmolGlc(-1); cell specific oxygen uptake and carbon dioxide production rates reached 216 and 226 mmol.CmolX(-1)·h(-1), respectively, and the respiratory quotient (QR) value varied from 1.1 to 1.7. This is the first time that kinetic parameters and yields are reported for D. geothermalis DSM-11302 grown on a mineral medium in well-controlled batch culture.

Quantitative Characterization of the Growth of Deinococcus geothermalis DSM-11302: Effect of Inoculum Size, Growth Medium and Culture Conditions

PubMed Central

Bornot, Julie; Molina-Jouve, Carole; Uribelarrea, Jean-Louis; Gorret, Nathalie

2015-01-01

Due to their remarkable resistance to extreme conditions, Deinococcaceae strains are of great interest to biotechnological prospects. However, the physiology of the extremophile strain Deinococcus geothermalis has scarcely been studied and is not well understood. The physiological behaviour was then studied in well-controlled conditions in flask and bioreactor cultures. The growth of D. geothermalis type strains was compared. Among the strains tested, the strain from the German Collection of Microorganisms (Deutsche Sammlung von Mikroorganismen DSM) DSM-11302 was found to give the highest biomass concentration and growth rate: in a complex medium with glucose, the growth rate reached 0.75 h−1 at 45 °C. Yeast extract concentration in the medium had significant constitutive and catalytic effects. Furthermore, the results showed that the physiological descriptors were not affected by the inoculum preparation steps. A batch culture of D. geothermalis DSM-11302 on defined medium was carried out: cells grew exponentially with a maximal growth rate of 0.28 h−1 and D. geothermalis DSM-11302 biomass reached 1.4 g·L−1 in 20 h. Then, 1.4 gDryCellWeight of biomass (X) was obtained from 5.6 g glucose (Glc) consumed as carbon source, corresponding to a yield of 0.3 CmolX·CmolGlc−1; cell specific oxygen uptake and carbon dioxide production rates reached 216 and 226 mmol.CmolX−1·h−1, respectively, and the respiratory quotient (QR) value varied from 1.1 to 1.7. This is the first time that kinetic parameters and yields are reported for D. geothermalis DSM-11302 grown on a mineral medium in well-controlled batch culture. PMID:27682099

Diazotrophic bacteria isolated from wild rice Oryza glumaepatula (Poaceae) in the Brazilian Amazon.

PubMed

Júnior, Paulo Ivan Fernandes; Pereira, Gilmara Maria Duarte; Perin, Liamara; da Silva, Luana Mesquita; Baraúna, Alexandre Cardoso; Alvess, Francilene Muniz; Passos, Samuel Ribeiro; Zilli, Jerri Edson

2013-06-01

The association of wild grasses with diazotrophic bacteria in Brazilian biomes is poorly understood. The isolation and characterization of bacteria associated with wild grasses can contribute to understand the diazotrophic ecology as well as to identify bacteria with biotechnological applications. In this study, we isolated and characterized diazotrophic bacterial isolates from Oryza glumaepatula collected in Cerrado and Forest areas of the Amazon in Roraima State, Brazil. Healthy O. glumepatula plants were collected at five sampling sites at Forest and seven at Cerrado, respectively. The plants were collected at the Cerrado areas in September 2008 while the Forest plants were collected in June/2008 and April/2009. The plants and the soil adhering to the roots were transferred to pots and grown for 35 days in greenhouse conditions. During the harvest, the shoots and the roots were crushed separately in a saline solution; the suspension was diluted serially and inoculated in Petri dishes containing Dyg's medium. All distinct bacterial colonies were purified in the same medium. The diazotrophic capacity of each bacterium in microaerophilic conditions was assessed in semisolid BMGM medium. In addition, the pellicles forming bacterial isolates were also evaluated by PCR amplification for nifH gene. The diversity of nifH bacteria was analyzed by Box-PCR fingerprinting. For selected strains, the growth promoting capacity of O. sativa as a model plant was also evaluated. A total of 992 bacterial isolates were obtained. Fifty-one bacteria were able to form pellicles in the semisolid medium and 38 also positively amplified the 360 bp nifH gene fragment. Among the 38 nifH+ isolates, 24 were obtained from the shoots, while 14 originated from the roots. The Box-PCR profiles showed that the bacterial isolates obtained in this study presented a low similarity with the reference strains belonging to the Herbaspirillum, Azospirillum and Burkholderia genus. The growth-promoting ability was confirmed for at least five isolates. For these bacteria, the root and shoot growing results showed higher increases when compared to those observed in plants inoculated with the evaluated reference strains. These results indicate that O. glumaepatula is colonized by a high diverse diazotrophic community in the Brazilian Amazon. Further investigations are now being carried out to determine the taxonomic positions of these isolates and their growth promoting mechanisms.

Electromagnetic wave propagation through a dielectric-chiral interface and through a chiral slab

NASA Technical Reports Server (NTRS)

Bassiri, S.; Papas, C. H.; Engheta, N.

1988-01-01

The reflection from and transmission through a semiinfinite chiral medium are analyzed by obtaining the Fresnel equations in terms of parallel- and perpendicular-polarized modes, and a comparison is made with results reported previously. The chiral medium is described electromagnetically by the constitutive relations D = (epsilon)E+i(gamma)B and H = i(gamma)E+(1/mu)B. The constants epsilon, mu and gamma are real and have values that are fixed by the size, the shape, and the spatial distribution of the elements that collectively compose the medium. The conditions are obtained for the total internal reflection of the incident wave from the interface and for the existence of the Brewster angle. The effects of the chirality on the polarization and the intensity of the reflected wave from the chiral half-space are discussed and illustrated by using the Stokes parameters. The propagation of electromagnetic wave through an infinite slab of chiral medium is formulated for oblique incidence and solved analytically for the case of normal incidence.

Collecting near mature and immature orchid seeds for ex situ conservation: 'in vitro collecting' as a case study.

PubMed

Kendon, Jonathan P; Rajaovelona, Landy; Sandford, Helen; Fang, Rui; Bell, Jake; Sarasan, Viswambharan

2017-08-08

Lack of phenological information and efficient collecting methods are considered impediments for orchid seed collecting. This leads to opportunistic collecting as part of general seed collecting schedules that may last few weeks especially in remote areas. The study explored the feasibility of collecting near mature and immature seeds to support conservation action plans. Mature, near mature and immature seeds of orchids were collected from the wild in the Central Highlands of Madagascar (CHM). Seed capsules were collected in sterile culture medium in the wild, to prevent deterioration of seeds inside the capsule after collecting, later to be cultured under laboratory conditions. Seed capsules collected by the in vitro collecting (IVC) method were kept in very good condition for up to 4 weeks before germination under in vitro conditions. Significantly faster and higher germination rate (p < 0.001) than mature seeds was observed in a number of taxa collected during a 3 year-long study. In some taxa even immature seeds, with no apparent sign of testa covering the embryo, germinated following IVC where mature seeds failed to germinate. We propose that IVC method has potential to complement conventional seed collecting by increasing the germplasm that can be used in integrated conservation action plans. Improvements can be made in developing collections for taxa from biodiversity hotspots and remote areas where collecting requires considerable resources. This method can further be used on a wider selection of plants from different geographic areas and on embryo rescue programmes for economically important plants.

New preservation method for mouse spermatozoa without freezing.

PubMed

Van Thuan, Nguyen; Wakayama, Sayaka; Kishigami, Satoshi; Wakayama, Teruhiko

2005-02-01

The objective of this study was to investigate the preservation of spermatozoa in a simple medium without freezing and to examine the effects of the preserved sperm on fertilization and development after injection into mature mouse oocytes. Mouse spermatozoa were collected from two caudae epididymides of mature B6D2F1 males and stored under various conditions: 1) in KSOMaa medium (potassium simplex optimized medium with amino acids) supplemented with 0, 1, or 4 mg/ml BSA and held at room temperature (RT, 27 degrees C); 2) in KSOMaa medium containing 4 mg/ml BSA (KSOM-BSA) and held at 4 degrees C, RT, or 37 degrees C (CO2 incubator); 3) in KSOM-BSA with osmolarity ranging from 271 to 2000 mOsmol, adjusted by addition of NaCl and held at 4 degrees C; and 4) a two-step preservation system consisting of storage in 800 mOsmol KSOM-BSA for 1 wk at RT followed by storage at -20 degrees C. Preservation of mouse spermatozoa at 4 degrees C in a medium with high osmolarity (700-1000 mOsmol) resulted in the highest frequency of live births after intracytoplasmic sperm injection (ICSI) into mature oocytes. The optimal conditions for preservation of mouse spermatozoa were 800 mOsmol KSOM containing 4 mg/ml BSA and a holding temperature of 4 degrees C. More than 40% of oocytes injected with sperm heads stored under these conditions for 2 mo developed to the morula/blastocyst stage in vitro and 39% of the embryos developed to term after transfer to recipient mice. Our results also indicate that mouse spermatozoa can be stored in 800 mOsmol KSOM-BSA medium at RT for 1 wk and then at -20 degrees C for up to 3 mo and retain their competence for ICSI. These new preservation methods permit extended conservation of viable spermatozoa that are capable of supporting normal embryonic development and the live birth of healthy offspring after ICSI.

Comparison of limb kinematics between collected and lengthened (medium/extended) trot in two groups of dressage horses on two different surfaces.

PubMed

Walker, V A; Tranquille, C A; Newton, J R; Dyson, S J; Brandham, J; Northrop, A J; Murray, R C

2017-09-01

Dressage horses are often asked to work in lengthened paces during training and competition, but to date there is limited information about the biomechanics of dressage-specific paces. Preliminary work has shown increased fetlock extension in extended compared with collected paces, but further investigation of the kinematic differences between collected, medium and extended trot in dressage horses is warranted. Investigation of the effect of collected vs. medium/extended trot on limb kinematics of dressage horses. Prospective kinematic evaluation. Twenty clinically sound horses in active dressage training were used. Group 1: Ten young horses (≤6 years) were assessed at collected and medium trot and Group 2: Ten mature horses (≥9 years) were assessed at collected and extended trot. All horses were evaluated on two different surfaces. High speed motion capture (240 Hz) was used to determine kinematic variables. Fore- and hindlimb angles were measured at mid-stance. Descriptive statistics and mixed effect multilevel regression analyses were performed. Speed and stride length were reduced and stride duration increased at collected compared with medium/extended trot. Lengthened trot (medium/extended trot) was associated with increased fetlock extension in both the fore- and hindlimbs in both groups of horses. Changes were greater in mature horses compared with young horses. Shoulder and carpus angles were associated with forelimb fetlock angle. Hock angle was not significantly influenced by pace. Surface had no effect on fetlock or hock angles. Only 2D motion analysis was carried out. Results may have differed in horses with more extreme gait characteristics. Medium/extended trot increases extension of the fore- and hindlimb fetlock joints compared with collected trot in both young and mature dressage horses, respectively. © 2017 EVJ Ltd.

Conditioned Medium from Placental Mesenchymal Stem Cells Reduces Oxidative Stress during the Cryopreservation of Ex Vivo Expanded Umbilical Cord Blood Cells.

PubMed

Kadekar, Darshana; Rangole, Sonal; Kale, Vaijayanti; Limaye, Lalita

2016-01-01

The limited cell dose in umbilical cord blood (UCB) necessitates ex vivo expansion of UCB. Further, the effective cryopreservation of these expanded cells is important in widening their use in the clinics. During cryopreservation, cells experience oxidative stress due to the generation of reactive oxygen species (ROS). Conditioned medium from mesenchymal stem cells (MSCs-CM) has been shown to alleviate the oxidative stress during wound healing, Alzheimer's disease and ischemic disease. This premise prompted us to investigate the influence of MSCs-CM during cryopreservation of expanded UCB cells. CM-was collected from cord/placental MSCs(C-MSCs-CM, P-MSC-CM). UCB CD34+cells were expanded as suspension cultures in serum free medium containing cytokines for 10 days. Cells were frozen with/without C-MSCs-CM and or P-MSCs-CM in the conventional freezing medium containing 20%FCS +10%DMSO using a programmable freezer and stored in liquid nitrogen. Upon revival, cells frozen with MSCs-CM were found to be superior to cells frozen in conventional medium in terms of viability, CD34+content and clonogenecity. Priming of revived cells for 48 hrs with MSCs-CM further improved their transplantation ability, as compared to those cultured without MSCs-CM. P-MSCs-CM radically reduced the oxidative stress in cryopreserved cells, resulting in better post thaw functionality of CD34+ cells than with C-MSCs-CM. The observed cryoprotective effect of MSCs-CM was primarily due to anti-oxidative and anti-apoptotic properties of the MSCs-CM and not because of the exosomes secreted by them. Our data suggest that MSCs-CM can serve as a valuable additive to the freezing or the priming medium for expanded UCB cells, which would increase their clinical applicability.

High-contrast 3D microscopic imaging of deep layers in a biological medium

NASA Astrophysics Data System (ADS)

Faridian, Ahmad; Pedrini, Giancarlo; Osten, Wolfgang

2014-03-01

Multilayer imaging of biological specimens is a demanding field of research, but scattering is one of the major obstacles in imaging the internal layers of a specimen. Although in many studies the biological object is assumed to be a weak scatterer, this condition is hardly satisfied for sub-millimeter sized organisms. The scattering medium is inhomogeneously distributed inside the specimen. Therefore, the scattering which occurs in the upper layers of a given internal layer of interest is different from the lower layers. That results in a different amount of collectable information for a specific point in the layer from each view. An opposed view dark-field digital holographic microscope (DHM) has been implemented in this work to collect the information concurrently from both views and increase the image quality. Implementing a DHM system gives the possibility to perform digital refocusing process and obtain multilayer images from each side without depth scanning of the object. The results have been presented and discussed here for a Drosophila embryo.

Radar Detection Performance in Medium Grazing Angle X-band Sea-clutter

DTIC Science & Technology

2015-12-01

polarisation HV: Horizontal transmit and Vertical receive polarisation IRSG: Imagery Radar Systems Group MAST06: Maritime Surveillance Trial 2006 PDF...different combinations of the polarisation, collection geometry and environmental conditions. Relevant models include the imaging radar systems group (IRSG...atmospheric and system losses respectively and pulse compression adds a gain given by the pulse length - bandwidth product, TpB. The thermal noise power in the

Gas analysis system for the Eight Foot High Temperature Tunnel

NASA Technical Reports Server (NTRS)

Leighty, Bradley D.; Davis, Patricia P.; Upchurch, Billy T.; Puster, Richard L.

1992-01-01

This paper describes the development of a gas collection and analysis system that is to be installed in the Eight-Foot High Temperature Tunnel (8' HTT) at NASA's Langley Research Center. This system will be used to analyze the test gas medium that results after burning a methane-air mixture to achieve the proper tunnel test parameters. The system consists of a sampling rake, a gas sample storage array, and a gas chromatographic system. Gas samples will be analyzed after each run to assure that proper combustion takes place in the tunnel resulting in a correctly balanced composition of the test gas medium. The proper ratio of gas species is critically necessary in order for the proper operation and testing of scramjet engines in the tunnel. After a variety of methane-air burn conditions have been analyzed, additional oxygen will be introduced into the combusted gas and the enriched test gas medium analyzed. The pre/post enrichment sets of data will be compared to verify that the gas species of the test gas medium is correctly balanced for testing of air-breathing engines.

Chitosan production by psychrotolerant Rhizopus oryzae in non-sterile open fermentation conditions.

PubMed

Tasar, Ozden Canli; Erdal, Serkan; Taskin, Mesut

2016-08-01

A new chitosan producing fungus was locally isolated from soil samples collected around Erzurum, Turkey and identified as Rhizopus oryzae PAS 17 (GenBank accession number KU318422.1). Cultivation in low cost non-sterile conditions was achieved by exploiting its ability to grow at low temperature and pH, thus, undesired microbial contamination could be eliminated when appropriate culture conditions (incubation temperature as 15°C and initial pH of the medium as 4.5) were selected. Medium composition and culture conditions were optimized using Taguchi orthogonal array (OA) design of experiment (DOE). An OA layout of L16 (4(5)) was constructed with five most influensive factors at four levels on chitosan production like, carbon source (molasses), metal ion (Mg(2+)), inoculum amount, agitation speed and incubation time. The optimal combinations of factors (molasses, 70ml/l; MgSO4·7H2O, 0.5g/l; inoculum, 6.7×10(6) spores/disc; agitation speed, 150rpm and incubation time, 8days) obtained from the proposed DOE methodology was further validated by analysis of variance (ANOVA) test and the results revealed the increment of chitosan and biomass yields of 14.45 and 8.58 folds from its unoptimized condition, respectively. Copyright © 2016 Elsevier B.V. All rights reserved.

A new pullulan-producing yeast and medium optimization for its exopolysaccharide production

NASA Astrophysics Data System (ADS)

Zhao, Shuangzhi; Chi, Zhenming

2003-04-01

Yeast strain Y68 producing high level of pullulan was isolated from the phyton collected in Toulouse, France. This strain was identified to be Rhodotorula bacarum by BIOLOG analysis. This is the first report that pullulan was produced by Rhodotorula bacarum. The optimal medium (g L-1) for pullulan production by this strain was 80 glucose, 20 soybean cake hydrolysate, 5 K2HPO4, 1 NaCl, 0.2 MgSO4·7H2O, 0.6 (NH4)2SO4, pH 7.0. Under this condition, 54 gL-1 pullulan was produced within 60 h at 30°C. Pullulan is a better starting material for producing marine prodrugs.

Chondrogenically primed tonsil-derived mesenchymal stem cells encapsulated in riboflavin-induced photocrosslinking collagen-hyaluronic acid hydrogel for meniscus tissue repairs.

PubMed

Koh, Rachel H; Jin, Yinji; Kang, Byung-Jae; Hwang, Nathaniel S

2017-04-15

Current meniscus tissue repairing strategies involve partial or total meniscectomy, followed by allograft transplantation or synthetic material implantation. However, allografts and synthetic implants have major drawbacks such as the limited supply of grafts and lack of integration into host tissue, respectively. In this study, we investigated the effects of conditioned medium (CM) from meniscal fibrochondrocytes and TGF-β3 on tonsil-derived mesenchymal stem cells (T-MSCs) for meniscus tissue engineering. CM-expanded T-MSCs were encapsulated in riboflavin-induced photocrosslinked collagen-hyaluronic acid (COL-RF-HA) hydrogels and cultured in chondrogenic medium containing TGF-β3. In vitro results indicate that CM-expanded cells followed by TGF-β3 exposure stimulated the expression of fibrocartilage-related genes (COL2, SOX9, ACAN, COL1) and production of extracellular matrix components. Histological assessment of in vitro and subcutaneously implanted in vivo constructs demonstrated that CM-expanded cells followed by TGF-β3 exposure resulted in highest cell proliferation, GAG accumulation, and collagen deposition. Furthermore, when implanted into meniscus defect model, CM treatment amplified the potential of TGF-β3 and induced complete regeneration. Conditioned medium derived from chondrocytes have been reported to effectively prime mesenchymal stem cells toward chondrogenic lineage. Type I collagen is the main component of meniscus extracellular matrix and hyaluronic acid is known to promote meniscus regeneration. In this manuscript, we investigated the effects of conditioned medium (CM) and transforming growth factor-β3 (TGF-β3) on tonsil-derived mesenchymal stem cells (T-MSCs) encapsulated in riboflavin-induced photocrosslinked collagen-hyaluronic acid (COL-RF-HA) hydrogel. We employed a novel source of conditioned medium, derived from meniscal fibrochondrocytes. Our in vitro and in vivo results collectively illustrate that CM-expanded cells followed by TGF-β3 exposure have the best potential for meniscus regeneration. This manuscript highlights a novel stem cell commitment strategy combined with biomaterials designs for meniscus regeneration. Copyright © 2017 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Stimuli-sensitive polymeric micelles as anticancer drug carriers.

PubMed

Na, Kun; Sethuraman, Vijay T; Bae, You Han

2006-11-01

Amphiphilic block copolymers often form core-shell type micelles by self-organization of the blocks in an aqueous medium or under specific experimental conditions. Polymeric micelles constructed from these polymers that contain a segment whose physical or chemical properties respond to small changes in environmental conditions are collectively called 'stimuli-sensitive' micelles. This class of nano-scaled constructs has been investigated as a promising anti-cancer drug carrier because the micelles are able to utilize small environmental changes and modify drug release kinetics, biodistribution and the interactions with tissues and cells. This review summarizes the recent progress in stimuli-sensitive micelles for tumor chemotherapy, particularly for those responding to hyperthermic conditions, tumor pH and endosomal/lysosomal pH.

A contradictory phenomenon of deshelving pulses in a dilute medium used for lengthened photon storage time.

PubMed

Ham, Byoung S

2010-08-16

Lengthening of photon storage time has been an important issue in quantum memories for long distance quantum communications utilizing quantum repeaters. Atom population transfer into an auxiliary spin state has been adapted to increase photon storage time of photon echoes. In this population transfer process phase shift to the collective atoms is inevitable, where the phase recovery condition must be multiple of 2pi to satisfy rephasing mechanism. Recent adaptation of the population transfer method to atomic frequency comb (AFC) echoes [Afzelius et al., Phys. Rev. Lett. 104, 040503 (2010)], where the population transfer method is originated in a controlled reversible inhomogeneous broadening technique [Moiseev and Kroll, Phys. Rev. Lett. 87, 173601 (2001)], however, shows contradictory phenomenon violating the phase recovery condition. This contradiction in AFC is reviewed as a general case of optical locking applied to a dilute medium for an optical depth-dependent coherence leakage resulting in partial retrieval efficiency.

Prospect of stem cell conditioned medium in regenerative medicine.

PubMed

Pawitan, Jeanne Adiwinata

2014-01-01

Stem cell-derived conditioned medium has a promising prospect to be produced as pharmaceuticals for regenerative medicine. To investigate various methods to obtain stem cell-derived conditioned medium (CM) to get an insight into their prospect of application in various diseases. Systematic review using keywords "stem cell" and "conditioned medium" or "secretome" and "therapy." Data concerning treated conditions/diseases, type of cell that was cultured, medium and supplements to culture the cells, culture condition, CM processing, growth factors and other secretions that were analyzed, method of application, and outcome were noted, grouped, tabulated, and analyzed. Most of CM using studies showed good results. However, the various CM, even when they were derived from the same kind of cells, were produced by different condition, that is, from different passage, culture medium, and culture condition. The growth factor yields of the various types of cells were available in some studies, and the cell number that was needed to produce CM for one application could be computed. Various stem cell-derived conditioned media were tested on various diseases and mostly showed good results. However, standardized methods of production and validations of their use need to be conducted.

Influence of socioeconomic aspects on lymphatic filariasis: A case-control study in Andhra Pradesh, India.

PubMed

Mutheneni, Srinivasa Rao; Upadhyayula, Suryanaryana Murty; Kumaraswamy, Sriram; Kadiri, Madhusudhan Rao; Nagalla, Balakrishna

2016-01-01

Lymphatic filariasis (LF) is a major public health problem in India. The objective of the study was to assess the impact of socioeconomic conditions on LF in Chittoor district of Andhra Pradesh, India. A survey was carried out from 2004 to 2007 during which, an epidemiological and socioeconomic data were collected and analysed. The microfilaria (mf) positive samples were taken as cases and matched with control group by sex and age (1:1) for case-control study. Bivariate and multivariate logistic regression was used to identify the potential risk factors for filariasis. Using principal component analysis (PCA), a socioeconomic index was developed and the data/scores were classified into low, medium and high categories. In total 5,133 blood smears were collected, of which 77 samples were found positive for microfilaria (1.52%). Multivariate analysis showed that the risk of filariasis was higher in groups of people with income < ₹1000 per month [OR = 2.752 (95%CI, 0.435-17.429)]; ₹ 1000-3000 per month [3.079 (0.923-0.275)]; people living in tiled house structure [1.641 (0.534-5.048)], with kutcha (uncemented) drainage system [19.427 (2.985- 126.410)], respondents who did not implemented mosquito avoidance measures [1.737 (0.563-5.358)]; and in people who were not aware about prevention and control of filariasis [1.042 (0.368-2.956)]. PCA showed that respondents with low (41.6%) and medium (33.8%) socioeconomic status are more prone to filariasis (p=0.036). The cross sectional study showed that the population with low and medium socioeconomic status are at higher risk of filariasis. The identified socioeconomic risk factors can be used as a guideline for improving the conditions for effective management of filariasis.

Mechanical loading prevents the stimulating effect of IL-1{beta} on osteocyte-modulated osteoclastogenesis

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

Kulkarni, Rishikesh N.; Bakker, Astrid D.; Everts, Vincent

Highlights: Black-Right-Pointing-Pointer Osteocyte incubation with IL-1{beta} stimulated osteocyte-modulated osteoclastogenesis. Black-Right-Pointing-Pointer Conditioned medium from IL-1{beta}-treated osteocytes increased osteoclastogenesis. Black-Right-Pointing-Pointer IL-1{beta} upregulated RANKL and downregulated OPG gene expression by osteocytes. Black-Right-Pointing-Pointer CYR61 is upregulated in mechanically stimulated osteocytes. Black-Right-Pointing-Pointer Mechanical loading of osteocytes may abolish IL-1{beta}-induced osteoclastogenesis. -- Abstract: Inflammatory diseases such as rheumatoid arthritis are often accompanied by higher plasma and synovial fluid levels of interleukin-1{beta} (IL-1{beta}), and by increased bone resorption. Since osteocytes are known to regulate bone resorption in response to changes in mechanical stimuli, we investigated whether IL-1{beta} affects osteocyte-modulated osteoclastogenesis in the presence or absence of mechanicalmore » loading of osteocytes. MLO-Y4 osteocytes were pre-incubated with IL-1{beta} (0.1-1 ng/ml) for 24 h. Cells were either or not subjected to mechanical loading by 1 h pulsating fluid flow (PFF; 0.7 {+-} 0.3 Pa, 5 Hz) in the presence of IL-1{beta} (0.1-1 ng/ml). Conditioned medium was collected after 1 h PFF or static cultures. Subsequently mouse bone marrow cells were seeded on top of the IL-1{beta}-treated osteocytes to determine osteoclastogenesis. Conditioned medium from mechanically loaded or static IL-1{beta}-treated osteocytes was added to co-cultures of untreated osteocytes and mouse bone marrow cells. Gene expression of cysteine-rich protein 61 (CYR61/CCN1), receptor activator of nuclear factor kappa-B ligand (RANKL), and osteoprotegerin (OPG) by osteocytes was determined immediately after PFF. Incubation of osteocytes with IL-1{beta}, as well as conditioned medium from static IL-1{beta}-treated osteocytes increased the formation of osteoclasts. However, conditioned medium from mechanically loaded IL-1{beta}-treated osteocytes prevented osteoclast formation. Incubation with IL-1{beta} upregulated RANKL and downregulated OPG gene expression by static osteocytes. PFF upregulated CYR61, RANKL, and OPG gene expression by osteocytes. Our results suggest that IL-1{beta} increases osteocyte-modulated osteoclastogenesis, and that mechanical loading of osteocytes may abolish IL-1{beta}-induced osteoclastogenesis.« less

Enhancing the auto-flocculation of photosynthetic bacteria to realize biomass recovery in brewery wastewater treatment.

PubMed

Lu, Haifeng; Dong, Shan; Zhang, Guangming; Han, Ting; Zhang, Yuanhui; Li, Baoming

2018-02-15

Photosynthetic bacteria (PSB) wastewater treatment technology can simultaneously realize wastewater purification and biomass production. The produced biomass contains high value-added products, which can be used in medical and agricultural industry. However, because of the small size and high electronegativity, PSB are hard to be collected from wastewater, which hampers the commercialization of PSB-based industrial processes. Auto-flocculation is a low cost, energy saving, non-toxic biomass collection method for microbiology. In this work, the influence factors with their optimal levels and mechanism for enhancing the auto-flocculation of PSB were investigated in pure cultivation medium. Then PSB auto-flocculation performance in real brewery wastewater was probed. Results showed that Na + concentration, pH and light intensity were three crucial factors except the initial inoculum sizes and temperature. In the pure medium cultivation system, the optimal condition for PSB auto-flocculation was as follows: pH was 9.5, inoculum size was 420 mg l -1 , Na + concentration was 0.067 mol l -1 , light intensity was 5000 lux, temperature was 30°C. Under the optimal condition, the auto-flocculation ratio and biomass recovery reached 85.0% and 1488 mg l -1 , which improved by 1.67-fold and 2.14-fold compared with the PSB enrichment cultivation conditions, respectively. Mechanism analysis showed that the protein/polysaccharides ratio and absolute Zeta potential value had a liner relationship. For the brewery wastewater treatment, under the above optimal condition, the chemical oxygen demand removal reached 94.3% with the auto-flocculation ratio and biomass recovery of 89.6% and 1510 mg l -1 , which increased 2.75-fold and 2.77-fold, respectively.

Cardiosphere-Derived Cells Facilitate Heart Repair by Modulating M1/M2 Macrophage Polarization and Neutrophil Recruitment

PubMed Central

Hasan, Al Shaimaa; Luo, Lan; Yan, Chen; Zhang, Tian-Xia; Urata, Yoshishige; Goto, Shinji; Mangoura, Safwat A.; Abdel-Raheem, Mahmoud H.; Zhang, Shouhua; Li, Tao-Sheng

2016-01-01

Cardiosphere-derived cells (CDCs), one of the promising stem cell sources for myocardial repair, have been tested in clinical trials and resulted in beneficial effects; however, the relevant mechanisms are not fully understood. In this study, we examined the hypothesis that CDCs favor heart repair by switching the macrophages from a pro-inflammatory phenotype (M1) into a regulatory anti-inflammatory phenotype (M2). Macrophages from mice were cultured with CDCs-conditioned medium or with fibroblasts-conditioned medium as a control. Immunostaining showed that CDCs-conditioned medium significantly enhanced the expression of CD206 (a marker for M2 macrophages), but decreased the expression of CD86 (a marker for M1 macrophages) 3 days after culture. For animal studies, we used an acute myocardial infarction model of mice. We injected CDCs, fibroblasts, or saline only into the border zone of infarction. Then we collected the heart tissues for histological analysis 5 and 14 days after treatment. Compared with control animals, CDCs treatment significantly decreased M1 macrophages and neutrophils but increased M2 macrophages in the infarcted heart. Furthermore, CDCs-treated mice had reduced infarct size and fewer apoptotic cells compared to the controls. Our data suggest that CDCs facilitate heart repair by modulating M1/M2 macrophage polarization and neutrophil recruitment, which may provide a new insight into the mechanisms of stem cell-based myocardial repair. PMID:27764217

Conditioned Medium from Placental Mesenchymal Stem Cells Reduces Oxidative Stress during the Cryopreservation of Ex Vivo Expanded Umbilical Cord Blood Cells

PubMed Central

Kadekar, Darshana; Rangole, Sonal; Kale, Vaijayanti; Limaye, Lalita

2016-01-01

Background The limited cell dose in umbilical cord blood (UCB) necessitates ex vivo expansion of UCB. Further, the effective cryopreservation of these expanded cells is important in widening their use in the clinics. During cryopreservation, cells experience oxidative stress due to the generation of reactive oxygen species (ROS). Conditioned medium from mesenchymal stem cells (MSCs-CM) has been shown to alleviate the oxidative stress during wound healing, Alzheimer’s disease and ischemic disease. This premise prompted us to investigate the influence of MSCs-CM during cryopreservation of expanded UCB cells. Methodology/Principle findings CM-was collected from cord/placental MSCs(C-MSCs-CM, P-MSC-CM). UCB CD34+cells were expanded as suspension cultures in serum free medium containing cytokines for 10 days. Cells were frozen with/without C-MSCs-CM and or P-MSCs-CM in the conventional freezing medium containing 20%FCS +10%DMSO using a programmable freezer and stored in liquid nitrogen. Upon revival, cells frozen with MSCs-CM were found to be superior to cells frozen in conventional medium in terms of viability, CD34+content and clonogenecity. Priming of revived cells for 48 hrs with MSCs-CM further improved their transplantation ability, as compared to those cultured without MSCs-CM. P-MSCs-CM radically reduced the oxidative stress in cryopreserved cells, resulting in better post thaw functionality of CD34+ cells than with C-MSCs-CM. The observed cryoprotective effect of MSCs-CM was primarily due to anti-oxidative and anti-apoptotic properties of the MSCs-CM and not because of the exosomes secreted by them. Conclusions/Significance Our data suggest that MSCs-CM can serve as a valuable additive to the freezing or the priming medium for expanded UCB cells, which would increase their clinical applicability. PMID:27780236

Determination of Root Exudates in a Steril Continuous Flow Culture. I. The Culture Method

PubMed Central

Richter, Martin; Wilms, Werner; Scheffer, Fritz

1968-01-01

A sterile plant culture consisting of culture vessels, culture solution container, collecting flasks for percolating nutrient solution, illumination and aeration systems and a suitable pump is described. Its difference with other culture methods is a very slow, continuous percolation of the nutrient solution through the rooting medium. Well defined and controllable conditions can thus be established in the rhizosphere over long culture periods. Samples can be collected at short intervals without disturbing the rhizosphere in any way nor endangering the sterility of the culture. One of the fundamental factors determining the special ecological characteristics of the plant rhizosphere is the liberation of organic and inorganic substances by the plant root. During the study of this phenomenon it became evident that the amount of substances liberated varies within wide limits (factors 100 to 1000) according to the conditions in which the root is developing. PMID:16656966

Germanium precipitation from collecting-mains liquor with tannin extract in an alkaline medium

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

Medvedev, K.P.; Mikhailov, N.F.; Petrapol'skaya, V.M.

1976-01-01

It is proposed to precipitate germanium in a slightly alkaline medium, with a neutral solution of tannin extract in aqueous alkali. The effects of various factors on germanium recovery from collecting-mains liquors have been studied.

Thermal processing of bone: in vitro response of mesenchymal cells to bone-conditioned medium.

PubMed

Sawada, K; Caballé-Serrano, J; Schuldt Filho, G; Bosshardt, D D; Schaller, B; Buser, D; Gruber, R

2015-08-01

The autoclaving, pasteurization, and freezing of bone grafts to remove bacteria and viruses, and for preservation, respectively, is considered to alter biological properties during graft consolidation. Fresh bone grafts release paracrine-like signals that are considered to support tissue regeneration. However, the impact of the autoclaving, pasteurization, and freezing of bone grafts on paracrine signals remains unknown. Therefore, conditioned medium was prepared from porcine cortical bone chips that had undergone thermal processing. The biological properties of the bone-conditioned medium were assessed by examining the changes in expression of target genes in oral fibroblasts. The data showed that conditioned medium obtained from bone chips that had undergone pasteurization and freezing changed the expression of adrenomedullin, pentraxin 3, BTB/POZ domain-containing protein 11, interleukin 11, NADPH oxidase 4, and proteoglycan 4 by at least five-fold in oral fibroblasts. Bone-conditioned medium obtained from autoclaved bone chips, however, failed to change the expression of the respective genes. Also, when bone-conditioned medium was prepared from fresh bone chips, autoclaving blocked the capacity of bone-conditioned medium to modulate gene expression. These in vitro results suggest that pasteurization and freezing of bone grafts preserve the release of biologically active paracrine signals, but autoclaving does not. Copyright © 2015 International Association of Oral and Maxillofacial Surgeons. Published by Elsevier Ltd. All rights reserved.

Biosynthesis of Diterpenoids in Tripterygium Adventitious Root Cultures1[OPEN

PubMed Central

Inabuy, Fainmarinat S.; Fischedick, Justin T.; Lange, Iris; Xu, Meimei

2017-01-01

Adventitious root cultures were developed from Tripterygium regelii, and growth conditions were optimized for the abundant production of diterpenoids, which can be collected directly from the medium. An analysis of publicly available transcriptome data sets collected with T. regelii roots and root cultures indicated the presence of a large gene family (with 20 members) for terpene synthases (TPSs). Nine candidate diterpene synthase genes were selected for follow-up functional evaluation, of which two belonged to the TPS-c, three to the TPS-e/f, and four to the TPS-b subfamilies. These genes were characterized by heterologous expression in a modular metabolic engineering system in Escherichia coli. Members of the TPS-c subfamily were characterized as copalyl diphosphate (diterpene) synthases, and those belonging to the TPS-e/f subfamily catalyzed the formation of precursors of kaurane diterpenoids. The TPS-b subfamily encompassed genes coding for enzymes involved in abietane diterpenoid biosynthesis and others with activities as monoterpene synthases. The structural characterization of diterpenoids accumulating in the medium of T. regelii adventitious root cultures, facilitated by searching the Spektraris online spectral database, enabled us to formulate a biosynthetic pathway for the biosynthesis of triptolide, a diterpenoid with pharmaceutical potential. Considering the significant enrichment of diterpenoids in the culture medium, fast-growing adventitious root cultures may hold promise as a sustainable resource for the large-scale production of triptolide. PMID:28751314

Hypoxic-Preconditioned Bone Marrow Stem Cell Medium Significantly Improves Outcome After Retinal Ischemia in Rats

PubMed Central

Roth, Steven; Dreixler, John C.; Mathew, Biji; Balyasnikova, Irina; Mann, Jacob R.; Boddapati, Venkat; Xue, Lai; Lesniak, Maciej S.

2016-01-01

Purpose We have previously demonstrated the protective effect of bone marrow stem cell (BMSC)-conditioned medium in retinal ischemic injury. We hypothesized here that hypoxic preconditioning of stem cells significantly enhances the neuroprotective effect of the conditioned medium and thereby augments the protective effect in ischemic retina. Methods Rats were subjected to retinal ischemia by increasing intraocular pressure to 130 to 135 mm Hg for 55 minutes. Hypoxic-preconditioned, hypoxic unconditioned, or normoxic medium was injected into the vitreous 24 hours after ischemia ended. Recovery was assessed 7 days after injections by comparing electroretinography measurements, histologic examination, and apoptosis (TUNEL, terminal deoxynucleotidyl transferase–mediated dUTP nick end labeling assay). To compare proteins secreted into the medium in the groups and the effect of hypoxic exposure, we used rat cytokine arrays. Results Eyes injected with hypoxic BMSC–conditioned medium 24 hours after ischemia demonstrated significantly enhanced return of retinal function, decreased retinal ganglion cell layer loss, and attenuated apoptosis compared to those administered normoxic or hypoxic unconditioned medium. Hypoxic-preconditioned medium had 21 significantly increased protein levels compared to normoxic medium. Conclusions The medium from hypoxic-preconditioned BMSCs robustly restored retinal function and prevented cell loss after ischemia when injected 24 hours after ischemia. The protective effect was even more pronounced than in our previous studies of normoxic conditioned medium. Prosurvival signals triggered by the secretome may play a role in this neuroprotective effect. PMID:27367588

Progranulin, a Major Secreted Protein of Mouse Adipose-Derived Stem Cells, Inhibits Light-Induced Retinal Degeneration

PubMed Central

Tsuruma, Kazuhiro; Yamauchi, Mika; Sugitani, Sou; Otsuka, Tomohiro; Ohno, Yuta; Nagahara, Yuki; Ikegame, Yuka; Shimazawa, Masamitsu; Yoshimura, Shinichi; Iwama, Toru

2014-01-01

Adipose tissue stromal vascular fraction contains mesenchymal stem cells, which show protective effects when administered to damaged tissues, mainly through secreted trophic factors. We examined the protective effects of adipose-derived stem cells (ASCs) and ASC-conditioned medium (ASC-CM) against retinal damage and identified the neuroprotective factors in ASC-CM. ASCs and mature adipocytes were isolated from mouse subcutaneous tissue. ASCs were injected intravitreally in a mouse model of light-induced retinal damage, and ASC injection recovered retinal function as measured by electroretinogram and inhibited outer nuclear layer, thinning, without engraftment of ASCs. ASC-CM and mature adipocyte-conditioned medium were collected after 72 hours of culture. In vitro, H2O2- and light-induced cell death was reduced in a photoreceptor cell line with ASC-CM but not with mature adipocyte-conditioned medium. In vivo, light-induced photoreceptor damage was evaluated by measurement of outer nuclear layer thickness at 5 days after light exposure and by electroretinogram recording. ASC-CM significantly inhibited photoreceptor degeneration and retinal dysfunction after light exposure. Progranulin was identified as a major secreted protein of ASCs that showed protective effects against retinal damage in vitro and in vivo. Furthermore, progranulin phosphorylated extracellular signal-regulated kinase, cAMP response element binding protein, and hepatocyte growth factor receptor, and protein kinase C signaling pathways were involved in the protective effects of progranulin. These findings suggest that ASC-CM and progranulin have neuroprotective effects in the light-induced retinal-damage model. Progranulin may be a potential target for the treatment of the degenerative diseases of the retina. PMID:24233842

Progranulin, a major secreted protein of mouse adipose-derived stem cells, inhibits light-induced retinal degeneration.

PubMed

Tsuruma, Kazuhiro; Yamauchi, Mika; Sugitani, Sou; Otsuka, Tomohiro; Ohno, Yuta; Nagahara, Yuki; Ikegame, Yuka; Shimazawa, Masamitsu; Yoshimura, Shinichi; Iwama, Toru; Hara, Hideaki

2014-01-01

Adipose tissue stromal vascular fraction contains mesenchymal stem cells, which show protective effects when administered to damaged tissues, mainly through secreted trophic factors. We examined the protective effects of adipose-derived stem cells (ASCs) and ASC-conditioned medium (ASC-CM) against retinal damage and identified the neuroprotective factors in ASC-CM. ASCs and mature adipocytes were isolated from mouse subcutaneous tissue. ASCs were injected intravitreally in a mouse model of light-induced retinal damage, and ASC injection recovered retinal function as measured by electroretinogram and inhibited outer nuclear layer, thinning, without engraftment of ASCs. ASC-CM and mature adipocyte-conditioned medium were collected after 72 hours of culture. In vitro, H2O2- and light-induced cell death was reduced in a photoreceptor cell line with ASC-CM but not with mature adipocyte-conditioned medium. In vivo, light-induced photoreceptor damage was evaluated by measurement of outer nuclear layer thickness at 5 days after light exposure and by electroretinogram recording. ASC-CM significantly inhibited photoreceptor degeneration and retinal dysfunction after light exposure. Progranulin was identified as a major secreted protein of ASCs that showed protective effects against retinal damage in vitro and in vivo. Furthermore, progranulin phosphorylated extracellular signal-regulated kinase, cAMP response element binding protein, and hepatocyte growth factor receptor, and protein kinase C signaling pathways were involved in the protective effects of progranulin. These findings suggest that ASC-CM and progranulin have neuroprotective effects in the light-induced retinal-damage model. Progranulin may be a potential target for the treatment of the degenerative diseases of the retina.

Screening of biofilm formation by beneficial vaginal lactobacilli and influence of culture media components.

PubMed

Terraf, M C Leccese; Juárez Tomás, M S; Nader-Macías, M E F; Silva, C

2012-12-01

To assess the ability of vaginal lactobacilli to form biofilm under different culture conditions and to determine the relationship between their growth and the capability of biofilm formation by selected strains. Fifteen Lactobacillus strains from human vagina were tested for biofilm formation by crystal violet staining. Only Lactobacillus rhamnosus Centro de Referencia para Lactobacilos Culture Collection (CRL) 1332, Lact. reuteri CRL 1324 and Lact. delbrueckii CRL 1510 were able to grow and form biofilm in culture media without Tween 80. However, Lact. gasseri CRL 1263 (a non-biofilm-forming strain) did not grow in these media. Scanning electron microscopy showed that Lact. rhamnosus CRL 1332 and Lact. reuteri CRL 1324 formed a highly structured biofilm, but only Lact. reuteri CRL 1324 showed a high amount of extracellular material in medium without Tween. Biofilm formation was significantly influenced by the strain, culture medium, inoculum concentration, microbial growth and chemical nature of the support used for the assay. The results allow the selection of biofilm-forming vaginal Lactobacillus strains and the conditions and factors that affect this phenomenon. © 2012 The Society for Applied Microbiology.

Work Ability and Its Related Factors Among Workers in Small and Medium Enterprises: Comparison Among Four ASEAN Countries.

PubMed

Kaewboonchoo, Orawan; Isahak, Marzuki; Susilowati, Indri; Phuong, Toai Nguyen; Morioka, Ikuharu; Harncharoen, Kitiphong; Low, Wah Yun; Ratanasiripong, Paul

2016-07-01

Work ability is related to many factors that might influence one's capacity to work. This study aimed to examine the work ability and its related factors among small and medium enterprises (SME) workers in 4 Association of Southeast Asian Nations (ASEAN) countries. The participants in this study included 2098 workers from food and textile industries in Indonesia, Malaysia, Thailand, and Vietnam. A cross-sectional survey of anonymous self-administrated questionnaire was designed to collect information on sociodemographic factors, work environment and ergonomic condition, musculoskeletal disorders, and work ability. Bivariate correlation coefficient and multiple linear regression analyses were used to predict the work ability. Results of this study confirm that work ability in 4 ASEAN countries was similar to that in European countries, and that the sociodemographic factors, work environment and ergonomic condition, and musculoskeletal disorder (MSD) were associated with work ability. These factors are important for considering occupational health and safety policy to promote work ability in food, textile, and other SME workers. © 2016 APJPH.

The impact of simulated microgravity on purinergic signaling in an endothelial and smooth muscle cell co-culture model

NASA Astrophysics Data System (ADS)

Zhang, Yu; Hemmersbach, Ruth; Lau, Patrick; Pansky, Andreas; Kassack, Matthias; Tobiasch, Edda

Astronauts suffer from cardiovascular deconditioning when they are exposed to microgravity conditions during space missions. Thus, current research focuses on the identification of the underlying mechanism also with respect to therapy and countermeasures. Endothelial cells (ECs) and smooth muscle cells (SMCs) play a key role in a variety of vascular functions. Gene expression, cytoskeleton morphology and apoptosis in both, ECs and SMCs, have shown alterations under simulated and real microgravity condition. However, all these data were observed during single culturing of either ECs or SMCs under microgravity conditions, which is different from the in vivo situation. Purinergic 2 (P2) receptors bind extracellular nucleotides and can regulate the vascular tone and vascular cell proliferation, migration and apoptosis. In this study primary ECs and SMCs were obtained from bovine aorta and characterized using specific markers. Here we show for the first time that the P2-receptor expressions pattern in ECs and in SMCs is altered after 24h in simulated microgravity. Specific receptors are down- or up-regulated on the gene and protein level. In addition the supernatant of ECs during culture was used as conditioned medium for SMCs and vice visa to investigate the influence of either cell type on the other. ECs and SMCs secret cytokines which induce pathogenic proliferation and an altered migration behavior under simulated microgravity conditions. Interestingly, co-culturing with condition medium could compensate this change. In detail, P2X7 was down-regulated in ECs after 24h clinorotation but recovered to the 1 g level when cultured with conditioned medium from SMCs collected under normal gravity. In conclusion, our data indicate that the paracrine effect between ECs and SMCs is an important regulator of cell behavior, also under altered gravity conditions. P2-receptor gene and protein expression were altered during microgravity. Since several P2-receptor artificial ligands are already established as drugs, P2-receptors might be a reasonable candidate for drug development for astronaut treatment of vascular deconditioning in the future. Keywords: simulated microgravity, purinergic signaling, endothelial cells, smooth muscle cells, co-culture, clinostat

Remediation of arsenic-contaminated soils and groundwaters

DOEpatents

Peters, Robert W.; Frank, James R.; Feng, Xiandong

1998-01-01

An in situ method for extraction of arsenic contaminants from a soil medium and remediation of the medium including contacting the medium with an extractant solution, directing the solution within and through the medium, and collecting the solution and contaminants. The method can also be used for arsenate and/or arsenite removal.

Decellularized bone extracellular matrix and human dental pulp stem cells as a construct for bone regeneration.

PubMed

Paduano, Francesco; Marrelli, Massimo; Alom, Noura; Amer, Mahetab; White, Lisa J; Shakesheff, Kevin M; Tatullo, Marco

2017-06-01

Dental pulp tissue represents a source of mesenchymal stem cells that have a strong differentiation potential towards the osteogenic lineage. The objective of the current study was to examine in vitro osteogenic induction of dental pulp stem cells (DPSCs) cultured on hydrogel scaffolds derived from decellularized bone extracellular matrix (bECM) compared to collagen type I (Col-I), the major component of bone matrix. DPSCs in combination with bECM hydrogels were cultured under three different conditions: basal medium, osteogenic medium and medium supplemented with growth factors (GFs) and cell growth, mineral deposition, gene and protein expression were investigated. The DPSCs/bECM hydrogel constructs cultured in basal medium showed that cells were viable after three weeks and that the expression of runt-related transcription factor 2 (RUNX-2) and bone sialoprotein (BSP) were significantly upregulated in the absence of extra osteogenic inducers compared to Col-I hydrogel scaffolds. In addition, the protein expression levels of BSP and osteocalcin were higher on bECM with respect to Col-I hydrogel scaffolds. Furthermore, DPSCs/bECM hydrogels cultured with osteogenic or GFs supplemented medium displayed a higher upregulation of the osteo-specific markers compared to Col-I hydrogels in identical media. Collectively, our results demonstrate that bECM hydrogels might be considered as suitable scaffolds to support osteogenic differentiation of DPSCs.

Feeder & basic fibroblast growth factor-free culture of human embryonic stem cells: Role of conditioned medium from immortalized human feeders.

PubMed

Teotia, Pooja; Sharma, Shilpa; Airan, Balram; Mohanty, Sujata

2016-12-01

Human embryonic stem cell (hESC) lines are commonly maintained on inactivated feeder cells, in the medium supplemented with basic fibroblast growth factor (bFGF). However, limited availability of feeder cells in culture, and the high cost of growth factors limit their use in scalable expansion of hESC cultures for clinical application. Here, we describe an efficient and cost-effective feeder and bFGF-free culture of hESCs using conditioned medium (CM) from immortalized feeder cells. KIND-1 hESC cell line was cultured in CM, collected from primary mouse embryonic fibroblast, human foreskin fibroblast (HFF) and immortalized HFF (I-HFF). Pluripotency of KIND-1 hESC cell line was confirmed by expression of genes, proteins and cell surface markers. In culture, these cells retained normal morphology, expressed all cell surface markers, could differentiate to embryoid bodies upon culture in vitro. Furthermore, I-HFF feeder cells without supplementation of bFGF released ample amount of endogenous bFGF to maintain stemness of hESC cells. The study results described the use of CM from immortalized feeder cells as a consistent source and an efficient, inexpensive feeder-free culture system for the maintenance of hESCs. Moreover, it was possible to maintain hESCs without exogenous supplementation of bFGF. Thus, the study could be extended to scalable expansion of hESC cultures for therapeutic purposes.

Fiber bundle phase conjugate mirror

DOEpatents

Ward, Benjamin G.

2012-05-01

An improved method and apparatus for passively conjugating the phases of a distorted wavefronts resulting from optical phase mismatch between elements of a fiber laser array are disclosed. A method for passively conjugating a distorted wavefront comprises the steps of: multiplexing a plurality of probe fibers and a bundle pump fiber in a fiber bundle array; passing the multiplexed output from the fiber bundle array through a collimating lens and into one portion of a non-linear medium; passing the output from a pump collection fiber through a focusing lens and into another portion of the non-linear medium so that the output from the pump collection fiber mixes with the multiplexed output from the fiber bundle; adjusting one or more degrees of freedom of one or more of the fiber bundle array, the collimating lens, the focusing lens, the non-linear medium, or the pump collection fiber to produce a standing wave in the non-linear medium.

Evidence of reversible bradycardia and arrhythmias caused by immunogenic proteins secreted by T. cruzi in isolated rat hearts.

PubMed

Rodríguez-Angulo, Héctor O; Toro-Mendoza, Jhoan; Marques, Juan A; Concepción, Juan L; Bonfante-Cabarcas, Rafael; Higuerey, Yoliver; Thomas, Luz E; Balzano-Nogueira, Leandro; López, José R; Mijares, Alfredo

2015-02-01

Chagas cardiomyopathy, caused by the protozoan Trypanosoma cruzi, is characterized by alterations in intracellular ion, heart failure and arrhythmias. Arrhythmias have been related to sudden death, even in asymptomatic patients, and their molecular mechanisms have not been fully elucidated. The aim of this study is to demonstrate the effect of proteins secreted by T. cruzi on healthy, isolated beating rat heart model under a non-damage-inducing protocol. We established a non-damage-inducing recirculation-reoxygenation model where ultrafiltrate fractions of conditioned medium control or conditioned infected medium were perfused at a standard flow rate and under partial oxygenation. Western blotting with chagasic patient serum was performed to determine the antigenicity of the conditioned infected medium fractions. We observed bradycardia, ventricular fibrillation and complete atrioventricular block in hearts during perfusion with >50 kDa conditioned infected culture medium. The preincubation of conditioned infected medium with chagasic serum abolished the bradycardia and arrhythmias. The proteins present in the conditioned infected culture medium of >50 kDa fractions were recognized by the chagasic patient sera associated with arrhythmias. These results suggest that proteins secreted by T. cruzi are involved in Chagas disease arrhythmias and may be a potential biomarker in chagasic patients.

Evidence of Reversible Bradycardia and Arrhythmias Caused by Immunogenic Proteins Secreted by T. cruzi in Isolated Rat Hearts

PubMed Central

Rodríguez-Angulo, Héctor O.; Toro-Mendoza, Jhoan; Marques, Juan A.; Concepción, Juan L.; Bonfante-Cabarcas, Rafael; Higuerey, Yoliver; Thomas, Luz E.; Balzano-Nogueira, Leandro; López, José R.; Mijares, Alfredo

2015-01-01

Rationale Chagas cardiomyopathy, caused by the protozoan Trypanosoma cruzi, is characterized by alterations in intracellular ion, heart failure and arrhythmias. Arrhythmias have been related to sudden death, even in asymptomatic patients, and their molecular mechanisms have not been fully elucidated. Objective The aim of this study is to demonstrate the effect of proteins secreted by T. cruzi on healthy, isolated beating rat heart model under a non-damage-inducing protocol. Methods and Results We established a non-damage-inducing recirculation-reoxygenation model where ultrafiltrate fractions of conditioned medium control or conditioned infected medium were perfused at a standard flow rate and under partial oxygenation. Western blotting with chagasic patient serum was performed to determine the antigenicity of the conditioned infected medium fractions. We observed bradycardia, ventricular fibrillation and complete atrioventricular block in hearts during perfusion with >50 kDa conditioned infected culture medium. The preincubation of conditioned infected medium with chagasic serum abolished the bradycardia and arrhythmias. The proteins present in the conditioned infected culture medium of >50 kDa fractions were recognized by the chagasic patient sera associated with arrhythmias. Conclusions These results suggest that proteins secreted by T. cruzi are involved in Chagas disease arrhythmias and may be a potential biomarker in chagasic patients. PMID:25647069

A simple quantitative approach for the determination of long and medium chain lipids in bio-relevant matrices by high performance liquid chromatography with refractive index detection.

PubMed

Lee, Kathy Wai Yu; Porter, Christopher J H; Boyd, Ben J

2013-09-01

There is increasing attention in the literature towards understanding the behaviour of lipid-based drug formulations under digestion conditions using in vitro and in vivo methods. This necessitates a convenient method for quantitation of lipids and lipid digestion products. In this study, a simple and accessible method for the separation and quantitative determination of typical formulation and digested lipids using high performance liquid chromatography coupled to refractive index detection (HPLC-RI) is described. Long and medium chain lipids were separated and quantified in a biological matrix (gastrointestinal content) without derivatisation using HPLC-RI on C18 and C8 columns, respectively. The intra- and inter-assay accuracy was between 92% and 106%, and the assays were precise to within a coefficient of variation of less than 10% over the range of 0.1-2 mg/mL for both long and medium chain lipids. This method is also shown to be suitable for quantifying the lipolysis products collected from the gastrointestinal tract in the course of in vivo lipid digestion studies.

Remediation of arsenic-contaminated soils and groundwaters

DOEpatents

Peters, R.W.; Frank, J.R.; Feng, X.

1998-06-23

An in situ method is described for extraction of arsenic contaminants from a soil medium and remediation of the medium including contacting the medium with an extractant solution, directing the solution within and through the medium, and collecting the solution and contaminants. The method can also be used for arsenate and/or arsenite removal. 8 figs.

Impact of Nutrient Restriction on the Structure of Listeria monocytogenes Biofilm Grown in a Microfluidic System.

PubMed

Cherifi, Tamazight; Jacques, Mario; Quessy, Sylvain; Fravalo, Philippe

2017-01-01

Biofilm formation by the pathogen Listeria monocytogenes is a major concern in food industries. The aim of this work was to elucidate the effect of nutrient limitation on both biofilm architecture and on the viability of the bacteria in microfluidic growth conditions. Biofilm formation by two L. monocytogenes strains was performed in a rich medium (BHI) and in a 10-fold diluted BHI (BHI/10) at 30°C for 24 h by using both static conditions and the microfluidic system Bioflux. In dynamic conditions, biofilms grown in rich and poor medium showed significant differences as well in structure and in the resulting biovolume. In BHI/10, biofilm was organized in a knitted network where cells formed long chains, whereas in the rich medium, the observed structure was homogeneous cellular multilayers. Biofilm biovolume production in BHI/10 was significantly higher than in BHI in these dynamic conditions. Interestingly, biovolume of dead cells in biofilms formed under limited nutrient conditions (BHI/10) was significantly higher than in biofilms formed in the BHI medium. In the other hand, in static conditions, biofilm is organized in a multilayer cells and dispersed cells in a rich medium BHI and poor medium BHI/10 respectively. There was significantly more biomass in the rich medium compared to BHI/10 but no difference was noted in the dead/damaged subpopulation showing how L. monocytogenes biofilm could be affected by the growth conditions. This work demonstrated that nutrient concentration affects biofilm structure and the proportion of dead cells in biofilms under microfluidic condition. Our study also showed that limited nutrients play an important role in the structural stability of L. monocytogenes biofilm by enhancing cell death and liberating extracellular DNA.

Impact of Nutrient Restriction on the Structure of Listeria monocytogenes Biofilm Grown in a Microfluidic System

PubMed Central

Cherifi, Tamazight; Jacques, Mario; Quessy, Sylvain; Fravalo, Philippe

2017-01-01

Biofilm formation by the pathogen Listeria monocytogenes is a major concern in food industries. The aim of this work was to elucidate the effect of nutrient limitation on both biofilm architecture and on the viability of the bacteria in microfluidic growth conditions. Biofilm formation by two L. monocytogenes strains was performed in a rich medium (BHI) and in a 10-fold diluted BHI (BHI/10) at 30°C for 24 h by using both static conditions and the microfluidic system Bioflux. In dynamic conditions, biofilms grown in rich and poor medium showed significant differences as well in structure and in the resulting biovolume. In BHI/10, biofilm was organized in a knitted network where cells formed long chains, whereas in the rich medium, the observed structure was homogeneous cellular multilayers. Biofilm biovolume production in BHI/10 was significantly higher than in BHI in these dynamic conditions. Interestingly, biovolume of dead cells in biofilms formed under limited nutrient conditions (BHI/10) was significantly higher than in biofilms formed in the BHI medium. In the other hand, in static conditions, biofilm is organized in a multilayer cells and dispersed cells in a rich medium BHI and poor medium BHI/10 respectively. There was significantly more biomass in the rich medium compared to BHI/10 but no difference was noted in the dead/damaged subpopulation showing how L. monocytogenes biofilm could be affected by the growth conditions. This work demonstrated that nutrient concentration affects biofilm structure and the proportion of dead cells in biofilms under microfluidic condition. Our study also showed that limited nutrients play an important role in the structural stability of L. monocytogenes biofilm by enhancing cell death and liberating extracellular DNA. PMID:28567031

Neutron detection apparatus and method

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

Derzon, Mark S.; Borek, III, Theodore T.

An apparatus for neutron detection is provided. The apparatus comprises a sensor medium in electrical contact with an electrode arrangement conformed to collect radiation-generated charge from the sensor medium. The sensor medium comprises a borazine and/or a borazine-based polymer.

Mesenchymal stem cell-conditioned medium triggers neuroinflammation and reactive species generation in organotypic cultures of rat hippocampus.

PubMed

Horn, Ana Paula; Bernardi, Andressa; Luiz Frozza, Rudimar; Grudzinski, Patrícia Bencke; Hoppe, Juliana Bender; de Souza, Luiz Fernando; Chagastelles, Pedro; de Souza Wyse, Angela Terezinha; Bernard, Elena Aida; Battastini, Ana Maria Oliveira; Campos, Maria Martha; Lenz, Guido; Nardi, Nance Beyer; Salbego, Christianne

2011-07-01

Cell therapy using bone marrow-derived mesenchymal stem cells (MSCs) seems to be a new alternative for the treatment of neurodegenerative diseases. Despite several promising results with their use, possible side effects are still unknown. In a previous work, we have shown that MSC-conditioned medium is toxic to hippocampal slice cultures and aggravates cell death induced by oxygen and glucose deprivation. In this work, we investigated whether the inflammatory response and/or reactive species formation could be involved in that toxicity. Rat organotypic hippocampal cultures were exposed for 24 h to conditioned medium from MSCs isolated from rat bone marrow. A marked glial activation was observed after exposure of cultures to MSC-conditioned medium, as evidenced by glial fibrillary acid protein (GFAP) and isolectin B(4) increase. Tumor necrosis factor-α and interleukin-6 levels were increased in the culture medium, and 2,7-dihydrodichlorofluorescein diacetate oxidation (indicating reactive species generation) and inducible nitric oxide synthase (iNOS) immunocontent were also higher after exposure of cultures to MSC-conditioned medium. Antioxidants (ascorbic acid and TROLOX(®)), N(ω)-nitro-l-arginine methyl ester hydrochloride, and anti-inflammatory drugs (indomethacin and dexamethasone) reduced cell death in hippocampal organotypic cultures after their exposure to MSC-conditioned medium. The results obtained here suggest that MSC-secreted factors trigger reactive species generation and neuroinflammation in organotypic cultures of hippocampus, introducing a note of caution in the use of these cells for neurological application.

Theory and Computation of Optimal Low- and Medium- Thrust Orbit Transfers

NASA Technical Reports Server (NTRS)

Goodson, Troy D.; Chuang, Jason C. H.; Ledsinger, Laura A.

1996-01-01

This report presents new theoretical results which lead to new algorithms for the computation of fuel-optimal multiple-burn orbit transfers of low and medium thrust. Theoretical results introduced herein show how to add burns to an optimal trajectory and show that the traditional set of necessary conditions may be replaced with a much simpler set of equations. Numerical results are presented to demonstrate the utility of the theoretical results and the new algorithms. Two indirect methods from the literature are shown to be effective for the optimal orbit transfer problem with relatively small numbers of burns. These methods are the Minimizing Boundary Condition Method (MBCM) and BOUNDSCO. Both of these methods make use of the first-order necessary conditions exactly as derived by optimal control theory. Perturbations due to Earth's oblateness and atmospheric drag are considered. These perturbations are of greatest interest for transfers that take place between low Earth orbit altitudes and geosynchronous orbit altitudes. Example extremal solutions including these effects and computed by the aforementioned methods are presented. An investigation is also made into a suboptimal multiple-burn guidance scheme. The FORTRAN code developed for this study has been collected together in a package named ORBPACK. ORBPACK's user manual is provided as an appendix to this report.

Situ soil sampling probe system with heated transfer line

DOEpatents

Robbat, Jr., Albert

2002-01-01

The present invention is directed both to an improved in situ penetrometer probe and to a heated, flexible transfer line. The line and probe may be implemented together in a penetrometer system in which the transfer line is used to connect the probe to a collector/analyzer at the surface. The probe comprises a heater that controls a temperature of a geologic medium surrounding the probe. At least one carrier gas port and vapor collection port are located on an external side wall of the probe. The carrier gas port provides a carrier gas into the geologic medium, and the collection port captures vapors from the geologic medium for analysis. In the transfer line, a flexible collection line that conveys a collected fluid, i.e., vapor, sample to a collector/analyzer. A flexible carrier gas line conveys a carrier gas to facilitate the collection of the sample. A system heating the collection line is also provided. Preferably the collection line is electrically conductive so that an electrical power source can generate a current through it so that the internal resistance generates heat.

On Research Methodology: The Cassette Tape as a Data Collection Medium.

ERIC Educational Resources Information Center

Yahya, Ismail bin; Moore, Gary E.

The efficiency and appropriateness of the cassette tape as a data collection medium was considered in this survey. A 19-item questionnaire was mailed to 117 agricultural teacher educators who had served at foreign locations. Most of the questions solicited qualitative data about their experiences, problems, and accomplishments. Respondents were…

Agreement for HPV genotyping detection between self-collected specimens on a FTA cartridge and clinician-collected specimens

PubMed Central

Guan, YaoYao; Gravitt, Patti E.; Howard, Roslyn; Eby, Yolanda J.; Wang, Shaoming; Li, Belinda; Feng, Changyan; Qiao, You-Lin; Castle, Philip E.

2016-01-01

The current method of transporting self-collected cervicovaginal specimen for HPV DNA testing relies on liquid based medium, which is challenging and expensive to transport. A novel, dry storage and transportation device, Whatman indicating FTA™ Elute Cartridge, avoids some of the pitfalls of liquid-based medium. This method has been shown to be comparable to liquid-based collection medium, but relative performance of self-collected (SC) and clinician-collected (CC) samples onto FTA cards has not been reported. The objective of this study is to compare the analytic performance of self- and clinician-collected samples onto FTA cartridges for the detection of carcinogenic HPV using Linear Array. There was a 91% agreement, 69% positive agreement, and kappa of 0.75 between the clinician-collected and self-collected specimens for detection of any carcinogenic HPV genotype. When the HPV results were categorized hierarchically according to cervical cancer risk, there was no difference in the distribution of the HPV results for the clinician- and self-collected specimens (p = 0.7). This study concludes that FTA elute cartridge is a promising method of specimen transport for cervical cancer screening programs considering using self-collected specimen and HPV testing. Larger studies with clinical endpoints are now needed to assess the clinical performance. PMID:23370404

Effect of potential oocyte transport protocols on blastocyst rates after intracytoplasmic sperm injection in the horse.

PubMed

Foss, R; Ortis, H; Hinrichs, K

2013-12-01

Intracytoplasmic sperm injection (ICSI) is used to produce foals from otherwise infertile mares and from stallions with limited sperm stores, but requires expensive equipment and is technically demanding. Methods to transport oocytes to ICSI laboratories would allow collection of oocytes by the referring veterinarian and enable greater application of this technique. This study was conducted to evaluate protocols that could be used to transport immature and maturing oocytes for ICSI. In vitro experiment. Oocytes were recovered by transvaginal ultrasound-guided follicular aspiration either from dominant follicles 24 h after deslorelin administration (dominant stimulated follicle [DSF]), or from subordinate (immature) follicles at the same time. To mimic transport, DSF oocytes were incubated overnight under differing conditions before ICSI; immature oocytes were placed in varying conditions overnight before in vitro maturation, followed by ICSI. The rate of blastocyst production was compared among treatments. Blastocysts were produced in all groups. Dominant stimulated follicle oocytes held in sealed tubes in pre-equilibrated control maturation medium maintained at 37°C yielded blastocyst development equal to that obtained for control incubated oocytes (70%). Dominant stimulated follicle oocytes held similarly in a warm passive device yielded poor blastocyst development (10%). Immature oocytes held for one or 2 nights in modified M199 medium, or for one night in commercial embryo holding solution, in air at room temperature, yielded 35-37% blastocyst development per injected oocyte. A commercially available medium can be used for shipping immature oocytes at room temperature with good resulting blastocyst rates. Better blastocyst rates per oocyte are obtained from DSF oocytes; however, these require maintenance at 37°C and as they are already maturing at the time of collection, are more sensitive to delays. This new, practical information supporting transport of both immature and DSF oocytes for ICSI may allow wider use of this procedure. © 2013 EVJ Ltd.

An improved medium for the anaerobic growth of Paracoccus denitrificans Pd1222

PubMed Central

Hahnke, Stefanie M.; Moosmann, Philipp; Erb, Tobias J.; Strous, Marc

2014-01-01

Paracoccus denitrificans is a well studied model organism with respect to its aerobic and anaerobic respiratory enzymes. However, until now, the growth medium for this organism has not been optimized for anaerobic growth. In particular, the requirements of P. denitrificans for trace elements (TEs) are not well known. In the present study we aimed to improve growth rates of P. denitrificans Pd1222 on a defined medium under anoxic conditions. We designed media containing different combinations of TEs at various concentrations, and tested their performance against previously reported media. Our results suggest that growth rate and yield depend on the availability and concentration of TEs in the medium. A chelated TE solution was more suitable than an acidified TE solution. Highest growth rates were achieved with medium comprising the TEs iron, manganese, molybdenum, copper and zinc ranging from 0.1 to 9 μM. On this medium, P. denitrificans Pd1222 grew with a generation time of 4.4 h under anoxic conditions and 2.8 h under oxic conditions. Diauxic growth was clearly shown with respect to nitrate and nitrite reduction under anoxic conditions. PMID:24550891

An improved medium for the anaerobic growth of Paracoccus denitrificans Pd1222.

PubMed

Hahnke, Stefanie M; Moosmann, Philipp; Erb, Tobias J; Strous, Marc

2014-01-01

Paracoccus denitrificans is a well studied model organism with respect to its aerobic and anaerobic respiratory enzymes. However, until now, the growth medium for this organism has not been optimized for anaerobic growth. In particular, the requirements of P. denitrificans for trace elements (TEs) are not well known. In the present study we aimed to improve growth rates of P. denitrificans Pd1222 on a defined medium under anoxic conditions. We designed media containing different combinations of TEs at various concentrations, and tested their performance against previously reported media. Our results suggest that growth rate and yield depend on the availability and concentration of TEs in the medium. A chelated TE solution was more suitable than an acidified TE solution. Highest growth rates were achieved with medium comprising the TEs iron, manganese, molybdenum, copper and zinc ranging from 0.1 to 9 μM. On this medium, P. denitrificans Pd1222 grew with a generation time of 4.4 h under anoxic conditions and 2.8 h under oxic conditions. Diauxic growth was clearly shown with respect to nitrate and nitrite reduction under anoxic conditions.

The Role of Paracrine and Autocrine Signaling in the Early Phase of Adipogenic Differentiation of Adipose-derived Stem Cells

PubMed Central

Hemmingsen, Mette; Vedel, Søren; Skafte-Pedersen, Peder; Sabourin, David; Collas, Philippe; Bruus, Henrik; Dufva, Martin

2013-01-01

Introduction High cell density is known to enhance adipogenic differentiation of mesenchymal stem cells, suggesting secretion of signaling factors or cell-contact-mediated signaling. By employing microfluidic biochip technology, we have been able to separate these two processes and study the secretion pathways. Methods and results Adipogenic differentiation of human adipose-derived stem cells (ASCs) cultured in a microfluidic system was investigated under perfusion conditions with an adipogenic medium or an adipogenic medium supplemented with supernatant from differentiating ASCs (conditioned medium). Conditioned medium increased adipogenic differentiation compared to adipogenic medium with respect to accumulation of lipid-filled vacuoles and gene expression of key adipogenic markers (C/EBPα, C/EBPβ, C/EBPδ, PPARγ, LPL and adiponectin). The positive effects of conditioned medium were observed early in the differentiation process. Conclusions Using different cell densities and microfluidic perfusion cell cultures to suppress the effects of cell-released factors, we have demonstrated the significant role played by auto- or paracrine signaling in adipocyte differentiation. The cell-released factor(s) were shown to act in the recruitment phase of the differentiation process. PMID:23723991

Human Milk Oligosaccharides Promote the Growth of Staphylococci

PubMed Central

Hunt, K. M.; Preuss, J.; Nissan, C.; Davlin, C. A.; Williams, J. E.; Shafii, B.; Richardson, A. D.; McGuire, M. K.; Bode, L.

2012-01-01

Human milk oligosaccharides (HMO), which constitute a major component of human milk, promote the growth of particular bacterial species in the infant's gastrointestinal tract. We hypothesized that HMO also interact with the bacterial communities present in human milk. To test this hypothesis, two experiments were conducted. First, milk samples were collected from healthy women (n = 16); culture-independent analysis of the bacterial communities was performed, HMO content was analyzed, and the relation between these factors was investigated. A positive correlation was observed between the relative abundance of Staphylococcus and total HMO content (r = 0.66). In a follow-up study, we conducted a series of in vitro growth curve experiments utilizing Staphylococcus aureus or Staphylococcus epidermidis and HMO isolated from human milk. HMO exhibited stimulatory effects on bacterial growth under various nutritional conditions. Analysis of culture supernatants from these experiments revealed that HMO did not measurably disappear from the culture medium, indicating that the growth-enhancing effects were not a result of bacterial metabolism of the HMO. Instead, stimulation of growth caused greater utilization of amino acids in minimal medium. Collectively, the data provide evidence that HMO may promote the growth of Staphylococcus species in the lactating mammary gland. PMID:22562995

Medium factors on anaerobic production of rhamnolipids by Pseudomonas aeruginosa SG and a simplifying medium for in situ microbial enhanced oil recovery applications.

PubMed

Zhao, Feng; Zhou, Jidong; Han, Siqin; Ma, Fang; Zhang, Ying; Zhang, Jie

2016-04-01

Aerobic production of rhamnolipid by Pseudomonas aeruginosa was extensively studied. But effect of medium composition on anaerobic production of rhamnolipid by P. aeruginosa was unknown. A simplifying medium facilitating anaerobic production of rhamnolipid is urgently needed for in situ microbial enhanced oil recovery (MEOR). Medium factors affecting anaerobic production of rhamnolipid were investigated using P. aeruginosa SG (Genbank accession number KJ995745). Medium composition for anaerobic production of rhamnolipid by P. aeruginosa is different from that for aerobic production of rhamnolipid. Both hydrophobic substrate and organic nitrogen inhibited rhamnolipid production under anaerobic conditions. Glycerol and nitrate were the best carbon and nitrogen source. The commonly used N limitation under aerobic conditions was not conducive to rhamnolipid production under anaerobic conditions because the initial cell growth demanded enough nitrate for anaerobic respiration. But rhamnolipid was also fast accumulated under nitrogen starvation conditions. Sufficient phosphate was needed for anaerobic production of rhamnolipid. SO4(2-) and Mg(2+) are required for anaerobic production of rhamnolipid. Results will contribute to isolation bacteria strains which can anaerobically produce rhamnolipid and medium optimization for anaerobic production of rhamnolipid. Based on medium optimization by response surface methodology and ions composition of reservoir formation water, a simplifying medium containing 70.3 g/l glycerol, 5.25 g/l NaNO3, 5.49 g/l KH2PO4, 6.9 g/l K2HPO4·3H2O and 0.40 g/l MgSO4 was designed. Using the simplifying medium, 630 mg/l of rhamnolipid was produced by SG, and the anaerobic culture emulsified crude oil to EI24 = 82.5 %. The simplifying medium was promising for in situ MEOR applications.

Avian Semen Collection by Cloacal Massage and Isolation of DNA from Sperm.

PubMed

Kucera, Aurelia C; Heidinger, Britt J

2018-02-05

Collection of semen may be useful for a wide range of applications including studies involving sperm quality, sperm telomere dynamics, and epigenetics. Birds are widely used subjects in biological research and are ideal for studies involving repeated sperm samples. However, few resources are currently available for those wishing to learn how to collect and extract DNA from avian sperm. Here we describe cloacal massage, a gentle, non-invasive manual technique for collecting avian sperm. Although this technique is established in the literature, it can be difficult to learn from the available descriptions. We also provide information for extracting DNA from avian semen using a commercial extraction kit with modifications. Cloacal massage can be easily used on any small- to medium-sized male bird in reproductive condition. Following collection, the semen can be used immediately for motility assays, or frozen for DNA extraction following the protocol described herein. This extraction protocol was refined for avian sperm and has been successfully used on samples collected from several passerine species (Passer domesticus, Spizella passerina, Haemorhous mexicanus, and Turdus migratorius) and one columbid (Columba livia).

Induction of endothelial cell proliferation by angiogenic factors released by activated monocytes

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

Pakala, Rajbabu; Watanabe, Takuya; Benedict, Claude R

2002-06-01

Introduction: Cell-cell interaction is an essential component of atherosclerotic plaque development. Activated monocytes appear to play a central role in the development of atherosclerosis, not only through foam cell formation but also via the production of various growth factors that induce proliferation of different cell types that are involved in the plaque development. Using serum free co-culture method, we determined the effect of monocytes on endothelial cell proliferation. Methods: Endothelial cell proliferation is determined by the amount of [{sup 3}H]thymidine incorporated in to the DNA. Basic fibroblast growth factor (b-FGF), vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8) levels inmore » the conditioned medium were determined by ELISA. Results: Conditioned medium from unactivated monocytes partially inhibited endothelial cell proliferation, whereas conditioned medium from activated monocytes promoted endothelial cell proliferation. The mitogenic effect of conditioned medium derived from activated monocytes is due to the presence of b-FGF, VEGF and IL-8. Neutralizing antibodies against b-FGF, VEGF and IL-8 partially reversed the mitogenic effect of conditioned medium derived from activated monocytes. When b-FGF, VEGF and IL-8 were immunoprecipitated from conditioned medium derived from activated monocytes, it is less mitogenic to endothelial cells. Conclusion: Activated monocytes may play an important role in the development of atherosclerotic plaque by producing endothelial cell growth factors.« less

wnt3a but not wnt11 supports self-renewal of embryonic stem cells

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

Singla, Dinender K.; Schneider, David J.; LeWinter, Martin M.

2006-06-30

wnt proteins (wnts) promote both differentiation of midbrain dopaminergic cells and self-renewal of haematopoietic stem cells. Mouse embryonic stem (ES) cells can be maintained and self-renew on mouse feeder cell layers or in media containing leukemia inhibitory factor (LIF). However, the effects of wnts on ES cells self-renewal and differentiation are not clearly understood. In the present study, we found that conditioned medium prepared from L cells expressing wnt3a can replace feeder cell layers and medium containing LIF in maintaining ES cells in the proliferation without differentiation (self-renewal) state. By contrast, conditioned medium from NIH3T3 cells expressing wnt11 did not.more » Alkaline phosphatase staining and compact colony formation were used as criteria of cells being in the undifferentiated state. ES cells maintained in medium conditioned by Wnt3a expressing cells underwent freezing and thawing while maintaining properties seen with LIF maintained ES cells. Purified wnt3a did not maintain self-renewal of ES cells for prolonged intervals. Thus, other factors in the medium conditioned by wnt3a expressing cells may have contributed to maintenance of ES cells in a self-renewal state. Pluripotency of ES cells was determined with the use of embryoid bodies in vitro. PD98059, a MEK specific inhibitor, promoted the growth of undifferentiated ES cells maintained in conditioned medium from wnt3a expressing cells. By contrast, the P38 MAPK inhibitor SB230580 did not, suggesting a role for the MEK pathway in self-renewal and differentiation of ES cells maintained in the wnt3a cell conditioned medium. Thus, our results show that conditioned medium from wnt3a but not wnt11 expressing cells can maintain ES cells in self-renewal and in a pluripotent state.« less

Evaluation of swabs and transport media for the recovery of Yersinia pestis.

PubMed

Gilbert, Sarah E; Rose, Laura J; Howard, Michele; Bradley, Meranda D; Shah, Sanjiv; Silvestri, Erin; Schaefer, Frank W; Noble-Wang, Judith

2014-01-01

The Government Accountability Office report investigating the surface sampling methods used during the 2001 mail contamination with Bacillus anthracis brought to light certain knowledge gaps that existed regarding environmental sampling with biothreat agents. Should a contamination event occur that involves non-spore forming biological select agents, such as Yersinia pestis, surface sample collection and processing protocols specific for these organisms will be needed. Two Y. pestis strains (virulent and avirulent), four swab types (polyester, macrofoam, rayon, and cotton), two pre-moistening solutions, six transport media, three temperatures, two levels of organic load, and four processing methods (vortexing, sonicating, combined sonicating and vortexing, no agitation) were evaluated to determine the conditions that would yield the highest percent of cultivable Y. pestis cells after storage. The optimum pre-moistening agent/transport media combination varied with the Y. pestis strain and swab type. Directly inoculated macrofoam swabs released the highest percent of cells into solution (93.9% recovered by culture) and rayon swabs were considered the second best swab option (77.0% recovered by culture). Storage at 4°C was found to be optimum for all storage times and transport media. In a worst case scenario, where the Y. pestis strain is not known and sample processing and analyses could not occur until 72h after sampling, macrofoam swabs pre-moistened with PBS supplemented with 0.05% Triton X-100 (PBSTX), stored at 4°C in neutralizing buffer (NB) as a transport medium (PBSTX/NB) or pre-moistened with NB and stored in PBSTX as a transport medium (NB/PBSTX), then vortexed 3min in the transport medium, performed significantly better than all other conditions for macrofoam swabs, regardless of strain tested (mean 12 - 72h recovery of 85.9-105.1%, p<0.001). In the same scenario, two combinations of pre-moistening medium/transport medium were found to be optimal for rayon swabs stored at 4°C (p<0.001), then sonicated 3min in the transport medium; PBSTX/PBSTX and NB/PBSTX (mean 12-72h recovery of 83.7-110.1%). © 2013.

[Massive multiplication of coffee (Coffee arabica L. cv. Catimor) through embryogenic cell suspension culture].

PubMed

Flermoso-Gallardo, L; Menóndez-Yuffá, A

2000-01-01

Cell suspensions offer several advantages as a system for massive propagation because of the high rates of multiplication, the higher homogeneity in the culture conditions and the possibility of automatization. In this study, different experimental conditions were analyzed to establish embryogenic cell suspension cultures of coffee. The best conditions to establish the embryogenic cell suspension cultures of coffee were as follows: coffee leaf sections were cultivated during 12 weeks (Stage I) in a solid medium with the Murashige and Skoog salts, 2 mg/l kinetin and 0.5 mg/l 2,4-dichlorophenoxiacetic acid (medium 1). Under these conditions the explants formed a callus tissue that was transferred to a liquid medium containing 5 mg/l of 6-benzylamlno-purine (medium 2). After 12 days in a shaking liquid medium (Stage II), the cultures were sieved and were maintained In the same media, which was renewed every eight days (Stage III). This method yielded 1884 embryos in 50 ml; placing the embryos under conditions for germination yielded plantlets of normal appearance.

Horizontal atmospheric turbulence, beam propagation, and modeling

NASA Astrophysics Data System (ADS)

Wilcox, Christopher C.; Santiago, Freddie; Martinez, Ty; Judd, K. Peter; Restaino, Sergio R.

2017-05-01

The turbulent effect from the Earth's atmosphere degrades the performance of an optical imaging system. Many studies have been conducted in the study of beam propagation in a turbulent medium. Horizontal beam propagation and correction presents many challenges when compared to vertical due to the far harsher turbulent conditions and increased complexity it induces. We investigate the collection of beam propagation data, analysis, and use for building a mathematical model of the horizontal turbulent path and the plans for an adaptive optical system to use this information to correct for horizontal path atmospheric turbulence.

Loss of the Liver X Receptors Disrupts the Balance of Hematopoietic Populations, With Detrimental Effects on Endothelial Progenitor Cells.

PubMed

Rasheed, Adil; Tsai, Ricky; Cummins, Carolyn L

2018-05-08

The liver X receptors (LXRs; α/β) are nuclear receptors known to regulate cholesterol homeostasis and the production of select hematopoietic populations. The objective of this study was to determine the importance of LXRs and a high-fat high-cholesterol diet on global hematopoiesis, with special emphasis on endothelial progenitor cells (EPCs), a vasoreparative cell type that is derived from bone marrow hematopoietic stem cells. Wild-type and LXR double-knockout ( Lxr αβ -/- ) mice were fed a Western diet (WD) to increase plasma cholesterol levels. In WD-fed Lxr αβ -/- mice, flow cytometry and complete blood cell counts revealed that hematopoietic stem cells, a myeloid progenitor, and mature circulating myeloid cells were increased; EPC numbers were significantly decreased. Hematopoietic stem cells from WD-fed Lxr αβ -/- mice showed increased cholesterol content, along with increased myeloid colony formation compared with chow-fed mice. In contrast, EPCs from WD-fed Lxr αβ -/- mice also demonstrated increased cellular cholesterol content that was associated with greater expression of the endothelial lineage markers Cd144 and Vegfr2 , suggesting accelerated differentiation of the EPCs. Treatment of human umbilical vein endothelial cells with conditioned medium collected from these EPCs increased THP-1 monocyte adhesion. Increased monocyte adhesion to conditioned medium-treated endothelial cells was recapitulated with conditioned medium from Lxr αβ -/- EPCs treated with cholesterol ex vivo, suggesting cholesterol is the main component of the WD inducing EPC dysfunction. LXRs are crucial for maintaining the balance of hematopoietic cells in a hypercholesterolemic environment and for mitigating the negative effects of cholesterol on EPC differentiation/secretome changes that promote monocyte-endothelial adhesion. © 2018 The Authors. Published on behalf of the American Heart Association, Inc., by Wiley.

An investigation into the stability of commercial versus MG63-derived hepatocyte growth factor under flow cultivation conditions.

PubMed

Meneghello, Giulia; Storm, Michael P; Chaudhuri, Julian B; De Bank, Paul A; Ellis, Marianne J

2015-03-01

The scale-up of tissue engineering cell culture must ensure that conditions are maintained while also being cost effective. Here we analyse the stability of hepatocyte growth factor (HGF) to investigate whether concentrations change under dynamic conditions, and compare commercial recombinant human HGF as an additive in 'standard medium', to HGF secreted by the osteosarcoma cell line MG63 as a 'preconditioned medium'. After 3 h under flow conditions, HGF in the standard medium degraded to 40% of its original concentration but HGF in the preconditioned medium remained at 100%. The concentration of secreted HGF was 10 times greater than the working concentration of commercially-available HGF. Thus HGF within this medium has increased stability; MG63-derived HGF should therefore be investigated as a cost-effective alternative to current lyophilised powders for use in in vitro models. Furthermore, we recommend that those intending to use HGF (or other growth factors) should consider similar stability testing before embarking on experiments with media flow.

Cost Factors in Scaling in SfM Collections and Processing Solutions

NASA Astrophysics Data System (ADS)

Cherry, J. E.

2015-12-01

In this talk I will discuss the economics of scaling Structure from Motion (SfM)-style collections from 1 km2 and below to 100's and 1000's of square kilometers. Considerations include the costs of the technical equipment: comparisons of small, medium, and large-format camera systems, as well as various GPS-INS systems and their impact on processing accuracy for various Ground Sampling Distances. Tradeoffs between camera formats and flight time are central. Weather conditions and planning high altitude versus low altitude flights are another economic factor, particularly in areas of persistently bad weather and in areas where ground logistics (i.e. hotel rooms and pilot incidentals) are expensive. Unique costs associated with UAS collections and experimental payloads will be discussed. Finally, the costs of equipment and labor differs in SfM processing than in conventional orthomosaic and LiDAR processing. There are opportunities for 'economies of scale' in SfM collections under certain circumstances but whether the accuracy specifications are firm/fixed or 'best effort' makes a difference.

Transformation of proanthocyanidin A2 to its isomers under different physiological pH conditions and common cell culture medium.

PubMed

Lu, Wen-Chien; Huang, Wei-Ting; Kumaran, Alaganandam; Ho, Chi-Tang; Hwang, Lucy Sun

2011-06-08

Proanthocyanidins constitute an important class of polyphenols ubiquitously found in plants. They have been extensively studied for their antioxidant capacity and bioactivity in vitro and in animal models. However, their stability under different pH conditions and in cell culture medium has not been well documented. In the present study, it was observed that proanthocyanidin A2 (PA2) was relatively more stable in acidic condition than in weak alkaline condition. PA2 was also quite unstable in basal-Dulbecco's Modified Eagle medium (b-DMEM medium) at 37 °C. The addition of PA2 to the cell culture medium accelerated its epimerization with a half-life of <15 min, and ethylenediaminetetraacetic acid (EDTA) could not stop the reaction. The results also demonstrated that the major isomers transformed in the weak alkaline condition or cell culture medium at 37 °C were identified as epicatechin-(4β→8; 2β→O→7)-ent-catechin (proanthocyanidin A4) and epicatechin-(4β→6; 2β→O→7)-ent-catechin. The rates of transformation were dependent on the pH or the components of the medium. Therefore, the results obtained for PA2 in the cell culture bioassays, which were usually carried out for 24 h, might not represent the true activity of the original PA2. The stability and transformation of PA2 should be considered when the bioactivity of PA2 is evaluated in a given cell culture system.

Evaluation of Volatile Organic Compounds and Carbonyl Compounds Present in the Cabins of Newly Produced, Medium- and Large-Size Coaches in China

PubMed Central

Lu, Yan-Yang; Lin, Yi; Zhang, Han; Ding, Dongxiao; Sun, Xia; Huang, Qiansheng; Lin, Lifeng; Chen, Ya-Jie; Chi, Yu-Lang; Dong, Sijun

2016-01-01

An air-conditioned coach is an important form of transportation in modern motorized society; as a result, there is an increasing concern of in-vehicle air pollution. In this study, we aimed to identify and quantify the levels of volatile organic compounds (VOCs) and carbonyl compounds (CCs) in air samples collected from the cabins of newly produced, medium- and large-size coaches. Among the identified VOCs and CCs, toluene, ethylbenzene, xylene, formaldehyde, acetaldehyde, acrolein/acetone, and isovaleraldehyde were relatively abundant in the cabins. Time was found to affect the emissions of the contaminants in the coaches. Except for benzaldehyde, valeraldehyde and benzene, the highest in-vehicle concentrations of VOCs and CCs were observed on the 15th day after coming off the assembly line, and the concentrations exhibited an approximately inverted U-shaped pattern as a function of time. Interestingly, this study also showed that the interior temperature of the coaches significantly affected the VOCs emissions from the interior materials, whereas the levels of CCs were mainly influenced by the relative humidity within the coaches. In China, guidelines and regulations for the in-vehicle air quality assessment of the coaches have not yet been issued. The results of this study provide further understanding of the in-vehicle air quality of air-conditioned coaches and can be used in the development of both specific and general rules regarding medium- and large-size coaches. PMID:27314375

Agreement for HPV genotyping detection between self-collected specimens on a FTA cartridge and clinician-collected specimens.

PubMed

Guan, Yaoyao; Gravitt, Patti E; Howard, Roslyn; Eby, Yolanda J; Wang, Shaoming; Li, Belinda; Feng, Changyan; Qiao, You-Lin; Castle, Philip E

2013-04-01

The current method of transporting self-collected cervicovaginal specimen for HPV DNA testing relies on liquid based medium, which is challenging and expensive to transport. A novel, dry storage and transportation device, Whatman indicating FTA™ Elute Cartridge, avoids some of the pitfalls of liquid-based medium. This method has been shown to be comparable to liquid-based collection medium, but relative performance of self-collected (SC) and clinician-collected (CC) samples onto FTA cards has not been reported. The objective of this study is to compare the analytic performance of self- and clinician-collected samples onto FTA cartridges for the detection of carcinogenic HPV using Linear Array. There was a 91% agreement, 69% positive agreement, and kappa of 0.75 between the clinician-collected and self-collected specimens for detection of any carcinogenic HPV genotype. When the HPV results were categorized hierarchically according to cervical cancer risk, there was no difference in the distribution of the HPV results for the clinician- and self-collected specimens (p=0.7). This study concludes that FTA elute cartridge is a promising method of specimen transport for cervical cancer screening programs considering using self-collected specimen and HPV testing. Larger studies with clinical endpoints are now needed to assess the clinical performance. Copyright © 2012 Elsevier B.V. All rights reserved.

Effect of culture medium, host strain and oxygen transfer on recombinant Fab antibody fragment yield and leakage to medium in shaken E. coli cultures.

PubMed

Ukkonen, Kaisa; Veijola, Johanna; Vasala, Antti; Neubauer, Peter

2013-07-29

Fab antibody fragments in E. coli are usually directed to the oxidizing periplasmic space for correct folding. From periplasm Fab fragments may further leak into extracellular medium. Information on the cultivation parameters affecting this leakage is scarce, and the unpredictable nature of Fab leakage is problematic regarding consistent product recovery. To elucidate the effects of cultivation conditions, we investigated Fab expression and accumulation into either periplasm or medium in E. coli K-12 and E. coli BL21 when grown in different types of media and under different aeration conditions. Small-scale Fab expression demonstrated significant differences in yield and ratio of periplasmic to extracellular Fab between different culture media and host strains. Expression in a medium with fed-batch-like glucose feeding provided highest total and extracellular yields in both strains. Unexpectedly, cultivation in baffled shake flasks at 150 rpm shaking speed resulted in higher yield and accumulation of Fabs into culture medium as compared to cultivation at 250 rpm. In the fed-batch medium, extracellular fraction in E. coli K-12 increased from 2-17% of total Fab at 250 rpm up to 75% at 150 rpm. This was partly due to increased lysis, but also leakage from intact cells increased at the lower shaking speed. Total Fab yield in E. coli BL21 in glycerol-based autoinduction medium was 5 to 9-fold higher at the lower shaking speed, and the extracellular fraction increased from ≤ 10% to 20-90%. The effect of aeration on Fab localization was reproduced in multiwell plate by variation of culture volume. Yield and leakage of Fab fragments are dependent on expression strain, culture medium, aeration rate, and the combination of these parameters. Maximum productivity in fed-batch-like conditions and in autoinduction medium is achieved under sufficiently oxygen-limited conditions, and lower aeration also promotes increased Fab accumulation into extracellular medium. These findings have practical implications for screening applications and small-scale Fab production, and highlight the importance of maintaining consistent aeration conditions during scale-up to avoid changes in product yield and localization. On the other hand, the dependency of Fab leakage on cultivation conditions provides a practical way to manipulate Fab localization.

Effect of culture medium, host strain and oxygen transfer on recombinant Fab antibody fragment yield and leakage to medium in shaken E. coli cultures

PubMed Central

2013-01-01

Background Fab antibody fragments in E. coli are usually directed to the oxidizing periplasmic space for correct folding. From periplasm Fab fragments may further leak into extracellular medium. Information on the cultivation parameters affecting this leakage is scarce, and the unpredictable nature of Fab leakage is problematic regarding consistent product recovery. To elucidate the effects of cultivation conditions, we investigated Fab expression and accumulation into either periplasm or medium in E. coli K-12 and E. coli BL21 when grown in different types of media and under different aeration conditions. Results Small-scale Fab expression demonstrated significant differences in yield and ratio of periplasmic to extracellular Fab between different culture media and host strains. Expression in a medium with fed-batch-like glucose feeding provided highest total and extracellular yields in both strains. Unexpectedly, cultivation in baffled shake flasks at 150 rpm shaking speed resulted in higher yield and accumulation of Fabs into culture medium as compared to cultivation at 250 rpm. In the fed-batch medium, extracellular fraction in E. coli K-12 increased from 2-17% of total Fab at 250 rpm up to 75% at 150 rpm. This was partly due to increased lysis, but also leakage from intact cells increased at the lower shaking speed. Total Fab yield in E. coli BL21 in glycerol-based autoinduction medium was 5 to 9-fold higher at the lower shaking speed, and the extracellular fraction increased from ≤ 10% to 20-90%. The effect of aeration on Fab localization was reproduced in multiwell plate by variation of culture volume. Conclusions Yield and leakage of Fab fragments are dependent on expression strain, culture medium, aeration rate, and the combination of these parameters. Maximum productivity in fed-batch-like conditions and in autoinduction medium is achieved under sufficiently oxygen-limited conditions, and lower aeration also promotes increased Fab accumulation into extracellular medium. These findings have practical implications for screening applications and small-scale Fab production, and highlight the importance of maintaining consistent aeration conditions during scale-up to avoid changes in product yield and localization. On the other hand, the dependency of Fab leakage on cultivation conditions provides a practical way to manipulate Fab localization. PMID:23895637

Effect of operating conditions in production of diagnostic Salmonella Enteritidis O-antigen-specific monoclonal antibody in different bioreactor systems.

PubMed

Ayyildiz-Tamis, Duygu; Nalbantsoy, Ayse; Elibol, Murat; Deliloglu-Gurhan, Saime Ismet

2014-01-01

In this study, different cultivation systems such as roller bottles (RB), 5-L stirred-tank bioreactor (STR), and disposable bioreactors were used to cultivate hybridoma for lab-scale production of Salmonella Enteritidis O-antigen-specific monoclonal antibody (MAb). Hybridoma cell line was cultivated in either serum-containing or serum-free medium (SFM) culture conditions. In STR, MAb production scaled up to 4 L, and production capabilities of the cells were also evaluated in different featured production systems. Moreover, the growth parameters of the cells in all production systems such as glucose consumption, lactate and ammonia production, and also MAb productivities were determined. Collected supernatants from the reactors were concentrated by a cross-flow filtration system. In conclusion, cells were not adapted to SFM in RB and STR. Therefore, less MAb titer in both STR and RB systems with SFM was observed compared to the cultures containing fetal bovine serum-supplemented medium. A higher MAb titer was gained in the membrane-aerated system compared to those in STR and RB. Although the highest MAb titer was obtained in the static membrane bioreactor system, the highest productivity was obtained in STR operated in semicontinuous mode with overlay aeration.

In vitro culture of coconut (Cocos nucifera L.) zygotic embryos.

PubMed

Engelmann, Florent; Malaurie, Bernard; N'Nan, Oulo

2011-01-01

Coconut is a very important crop for millions of people in tropical countries. With coconut, in vitro culture protocols have been developed with two main objectives, viz. the large scale production of particular types of coconuts and the international exchange and conservation of coconut germplasm. The methods described in this chapter have been developed in the framework of collaborative activities between research institutes in Côte d'Ivoire and France. Two coconut embryo in vitro collecting protocols have been established, one consisting of storing the disinfected embryos in a KCl solution until they are brought back to the laboratory, where they are re-disinfected and inoculated in vitro under sterile conditions, and the other including in vitro inoculation of the embryos in the field. For international germplasm exchange, zygotic embryos inoculated in vitro in plastic test tubes or endosperm cylinders containing embryos in plastic bags are used. For in vitro culture, embryos are inoculated on semi-solid medium supplemented with sucrose and activated charcoal and placed in the dark, and then transferred to light conditions with the same (solid or liquid) medium once the first true leaf is visible and the root system has started developing.

Conditioned medium from the three-dimensional culture of human umbilical cord perivascular cells accelerate the migration and proliferation of human keratinocyte and fibroblast.

PubMed

Kim, Min Ho; Wu, Wen Hao; Choi, Jee Hyun; Kim, Ji Hyun; Hong, Seok-Ho; Jun, Jin Hyun; Ko, Yong; Lee, Jong Hun

Previous studies have reported that the conditioned medium (CM) of bone marrow-mesenchymal stem cells (BM-MSCs) stimulate the migration and proliferation of cell types involved in the wound healing process. However, these studies only show MSC-CM effects that were obtained using a two-dimensional (2D) culture. Recently, a three-dimensional (3D) culture has been considered to be a more physiologically appropriate system than the 2D culture. In addition, it has been shown that the procurement of BM-MSC is invasive, and other sources of MSC are thus being explored. Recently, perivascular cells (PVCs) have been considered as an alternative source of cells for dermal wound healing. Therefore, in this study, a PVC-conditioned medium (CM) was collected from a 3D culture (PVC-CM-3D) using highly porous polystyrene-based membranes and compared with PVC-CM from a 2D culture (PVC-CM-2D) to investigate the effects on the migration and proliferation of human keratinocytes and fibroblasts. Moreover, the PVC-CM components from the 2D and 3D cultures were identified using 2D gel electrophoresis. The migrations of the keratinocytes cells and fibroblasts were significantly higher with PVC-CM-3D than with the 2D culture; similarly, the proliferation of keratinocytes was also highly stimulated by PVC-CM-3D. Proteomic analyses of the PVC-CM revealed that type I collagen was highly expressed in the 3D-culture system. Microtubule-actin cross-linked factor 1 (KIAA0465), nebulin-related anchoring protein, and thioredoxin were specifically expressed only in PVC-CM-3D. In addition, more EVs could be isolated from the PVC-CM-3D, and EVs were found to stimulate keratinocyte migration. Taken together, 3D-culture using a polystyrene scaffold is demonstrated to be a better system for providing better physiological conditions; therefore, PVC-CM-3D could be a promising option for skin-wound healing.

Identification and Evaluation of Reliable Reference Genes in the Medicinal Fungus Shiraia bambusicola.

PubMed

Song, Liang; Li, Tong; Fan, Li; Shen, Xiao-Ye; Hou, Cheng-Lin

2016-04-01

The stability of reference genes plays a vital role in real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis, which is generally regarded as a convenient and sensitive tool for the analysis of gene expression. A well-known medicinal fungus, Shiraia bambusicola, has great potential in the pharmaceutical, agricultural and food industries, but its suitable reference genes have not yet been determined. In the present study, 11 candidate reference genes in S. bambusicola were first evaluated and validated comprehensively. To identify the suitable reference genes for qRT-PCR analysis, three software-based algorithms, geNorm, NormFinder and Best Keeper, were applied to rank the tested genes. RNA samples were collected from seven fermentation stages using different media (potato dextrose or Czapek medium) and under different light conditions (12-h light/12-h dark and all-dark). The three most appropriate reference genes, ubi, tfc and ags, were able to normalize the qRT-PCR results under the culturing conditions of 12-h light/12-h dark, whereas the other three genes, vac, gke and acyl, performed better in the culturing conditions of all-dark growth. Therefore, under different light conditions, at least two reference genes (ubi and vac) could be employed to assure the reliability of qRT-PCR results. For both the natural culture medium (the most appropriate genes of this group: ubi, tfc and ags) and the chemically defined synthetic medium (the most stable genes of this group: tfc, vac and ef), the tfc gene remained the best gene used for normalizing the gene expression found with qRT-PCR. It is anticipated that these results would improve the selection of suitable reference genes for qRT-PCR assays and lay the foundation for an accurate analysis of gene expression in S. bambusicola.

Collective Lamb Shift of a Nanoscale Atomic Vapor Layer within a Sapphire Cavity

NASA Astrophysics Data System (ADS)

Peyrot, T.; Sortais, Y. R. P.; Browaeys, A.; Sargsyan, A.; Sarkisyan, D.; Keaveney, J.; Hughes, I. G.; Adams, C. S.

2018-06-01

We measure the near-resonant transmission of light through a dense medium of potassium vapor confined in a cell with nanometer thickness in order to investigate the origin and validity of the collective Lamb shift. A complete model including the multiple reflections in the nanocell reproduces accurately the observed line shape. It allows the extraction of a density-dependent shift and width of the bulk atomic medium resonance, deconvolved from the cavity effect. We observe an additional, unexpected dependence of the shift with the thickness of the medium. This extra dependence demands further experimental and theoretical investigations.

Antitumor Activity of Rat Mesenchymal Stem Cells during Direct or Indirect Co-Culturing with C6 Glioma Cells.

PubMed

Gabashvili, A N; Baklaushev, V P; Grinenko, N F; Mel'nikov, P A; Cherepanov, S A; Levinsky, A B; Chehonin, V P

2016-02-01

The tumor-suppressive effect of rat mesenchymal stem cells against low-differentiated rat C6 glioma cells during their direct and indirect co-culturing and during culturing of C6 glioma cells in the medium conditioned by mesenchymal stem cells was studied in an in vitro experiment. The most pronounced antitumor activity of mesenchymal stem cells was observed during direct co-culturing with C6 glioma cells. The number of live C6 glioma cells during indirect co-culturing and during culturing in conditioned medium was slightly higher than during direct co-culturing, but significantly differed from the control (C6 glioma cells cultured in medium conditioned by C6 glioma cells). The cytotoxic effect of medium conditioned by mesenchymal stem cells was not related to medium depletion by glioma cells during their growth. The medium conditioned by other "non-stem" cells (rat astrocytes and fibroblasts) produced no tumor-suppressive effect. Rat mesenchymal stem cells, similar to rat C6 glioma cells express connexin 43, the main astroglial gap junction protein. During co-culturing, mesenchymal stem cells and glioma C6 cells formed functionally active gap junctions. Gap junction blockade with connexon inhibitor carbenoxolone attenuated the antitumor effect observed during direct co-culturing of C6 glioma cells and mesenchymal stem cells to the level produced by conditioned medium. Cell-cell signaling mediated by gap junctions can be a mechanism of the tumor-suppressive effect of mesenchymal stem cells against C6 glioma cells. This phenomenon can be used for the development of new methods of cell therapy for high-grade malignant gliomas.

Aerobic Reduction of Arsenate by a Bacterium Isolated From Activated Sludge

NASA Astrophysics Data System (ADS)

Kozai, N.; Ohnuki, T.; Hanada, S.; Nakamura, K.; Francis, A. J.

2006-12-01

Microlunatus phosphovorus strain NM-1 is a polyphosphate-accumulating bacterium isolated from activated sludge. This bacterium takes up a large amount of polyphosphate under aerobic conditions and release phosphate ions by hydrolysis of polyphosphate to orthophosphate under anaerobic conditions to derive energy for taking up substrates. To understand the nature of this strain, especially, influence of potential contaminants in sewage and wastewater on growth, we have been investigating behavior of this bacterium in media containing arsenic. The present paper mainly reports reduction of arsenate by this bacterium under aerobic conditions. The strain NM-1 (JCM 9379) was aerobically cultured at 30 °C in a nutrient medium containing 2.5 g/l peptone, 0.5 g/l glucose, 1.5 g/l yeast extract, and arsenic [Na2HAsO4 (As(V)) or Na3AsO3 (As(III))] at concentrations between 0 and 50 mM. The cells collected from arsenic-free media were dispersed in buffer solutions containing 2mM HEPES, 10mM NaCl, prescribed concentrations of As(V), and 0-0.2 percent glucose. Then, this cell suspension was kept at 20 °C under aerobic or anaerobic conditions. The speciation of arsenic was carried out by ion chromatography and ICP-MS. The growth of the strain under aerobic conditions was enhanced by the addition of As(V) at the concentration between 1 and 10 mM. The maximum optical density of the culture in the medium containing 5mM As(V) was 1.4 times greater than that of the control culture. Below the As(V) concentration of 10mM, most of the As(V) was reduced to As(III). The growth of the strain under anaerobic conditions has not been observed so far. The cells in the buffer solutions reduced As(V) under aerobic condition. The reduction was enhanced by the addition of glucose. However, the cell did not reduce As(V) under anaerobic conditions. The strain NM-1 showed high resistance to As(V) and As(III). The maximum optical density of the culture grown in a medium containing 50 mM As(V) was only 20 percent lower than that of the control culture.

Atrazine degradation in a small stream in Iowa

USGS Publications Warehouse

Kolpin, D.W.; Kalkhoff, S.J.

1993-01-01

A study was conducted during 1990 through an 11.2-km reach of Roberts Creek in northeastern Iowa to determine the fate of atrazine in a surface water environment Water samples were collected at ~1-month intervals from April through November during stable low to medium flow conditions and analyzed for atrazine and two of its initial biotic degradation products, desethylatrazine and deisopropylatrazine. Samples were collected on the basis of a Lagrangian model of streamflow in order to sample the same parcel of water as it moved downstream. Atrazine concentrations substantially decreased (roughly 25-60%) between water entering and exiting the study reach during four of the seven sampling periods. During these same four sampling periods, the concentrations of the two biotic atrazine degradation products were constant or decreasing downstream, suggesting an abiotic degradation process.

REVIEWS OF TOPICAL PROBLEMS The quark-gluon medium

NASA Astrophysics Data System (ADS)

Dremin, Igor M.; Leonidov, Andrei V.

2011-02-01

The properties of the quark-gluon medium observed in high-energy nucleus-nucleus collisions are discussed. The main experimental facts about these collisions are briefly described and compared with data about proton-proton collisions. Both microscopic and macroscopic approaches to their description are reviewed. The chromodynamics of the quark-gluon medium at high energies is mainly considered. The energy loss of partons moving in this medium is treated. The principal conclusion is that the medium possesses some collective properties which are crucial for understanding the experimental observations.

Physical Forces Shape Group Identity of Swimming Pseudomonas putida Cells.

PubMed

Espeso, David R; Martínez-García, Esteban; de Lorenzo, Víctor; Goñi-Moreno, Ángel

2016-01-01

The often striking macroscopic patterns developed by motile bacterial populations on agar plates are a consequence of the environmental conditions where the cells grow and spread. Parameters such as medium stiffness and nutrient concentration have been reported to alter cell swimming behavior, while mutual interactions among populations shape collective patterns. One commonly observed occurrence is the mutual inhibition of clonal bacteria when moving toward each other, which results in a distinct halt at a finite distance on the agar matrix before having direct contact. The dynamics behind this phenomenon (i.e., intolerance to mix in time and space with otherwise identical others) has been traditionally explained in terms of cell-to-cell competition/cooperation regarding nutrient availability. In this work, the same scenario has been revisited from an alternative perspective: the effect of the physical mechanics that frame the process, in particular the consequences of collisions between moving bacteria and the semi-solid matrix of the swimming medium. To this end, we set up a simple experimental system in which the swimming patterns of Pseudomonas putida were tested with different geometries and agar concentrations. A computational analysis framework that highlights cell-to-medium interactions was developed to fit experimental observations. Simulated outputs suggested that the medium is compressed in the direction of the bacterial front motion. This phenomenon generates what was termed a compression wave that goes through the medium preceding the swimming population and that determines the visible high-level pattern. Taken together, the data suggested that the mechanical effects of the bacteria moving through the medium created a factual barrier that impedes to merge with neighboring cells swimming from a different site. The resulting divide between otherwise clonal bacteria is thus brought about by physical forces-not genetic or metabolic programs.

Laser-assisted surface modification of Ti-implant in air and water environment

NASA Astrophysics Data System (ADS)

Trtica, M.; Stasic, J.; Batani, D.; Benocci, R.; Narayanan, V.; Ciganovic, J.

2018-01-01

A study of the surface modification of titanium CP grade 2 implant/target with high intensity picosecond (Nd:YAG) laser, operating at 1064 nm wavelength and pulse duration of 40 ps, in gaseous (air) and liquid (water) medium, is presented. The exposure of Ti to a laser pulse energy of 17 mJ in both media - gaseous and liquid, induced specific surface features and phenomena: (i) enhancement of the implant surface roughness (higher in water). In this context, the damage depth is more prominent in water (as high as ∼40 μm) vs. air (∼14 μm). Also, the appearance of laser induced periodic surface structures (LIPSS) is recorded in both media, at periphery area, while in water they are registered at lower pulse count; (ii) variation of chemical surface content depending on the applied medium. Thus, in the central irradiation region, the oxygen was absent in air while its concentration was relatively high (6.44 wt%) in case of water; (iii) possibility of direct collection of synthesized titanium based nanoparticles in water environment, and (iv) formation of the plasma above the sample in both mediums, more volumetrically confined in water. These investigations showed that surface structuring and observed phenomena are in strong correlation with the medium used. The liquid - water seems like the medium of choice in regard to titanium implant biocompatibility and bio-activity (the water is a favorable medium for build-up of the oxide layer which affects bioactivity). The process of laser interaction with titanium implant targets was accompanied by the formation of plasma plume, which provides the additional sterilizing effect facilitating contaminant-free conditions.

Method and apparatus for high-efficiency direct contact condensation

DOEpatents

Bharathan, D.; Parent, Y.; Hassani, A.V.

1999-07-20

A direct contact condenser having a downward vapor flow chamber and an upward vapor flow chamber, wherein each of the vapor flow chambers includes a plurality of cooling liquid supplying pipes and a vapor-liquid contact medium disposed thereunder to facilitate contact and direct heat exchange between the vapor and cooling liquid. The contact medium includes a plurality of sheets arranged to form vertical interleaved channels or passageways for the vapor and cooling liquid streams. The upward vapor flow chamber also includes a second set of cooling liquid supplying pipes disposed beneath the vapor-liquid contact medium which operate intermittently in response to a pressure differential within the upward vapor flow chamber. The condenser further includes separate wells for collecting condensate and cooling liquid from each of the vapor flow chambers. In alternate embodiments, the condenser includes a cross-current flow chamber and an upward flow chamber, a plurality of upward flow chambers, or a single upward flow chamber. The method of use of the direct contact condenser of this invention includes passing a vapor stream sequentially through the downward and upward vapor flow chambers, where the vapor is condensed as a result of heat exchange with the cooling liquid in the contact medium. The concentration of noncondensable gases in the resulting condensate-liquid mixtures can be minimized by controlling the partial pressure of the vapor, which depends in part upon the geometry of the vapor-liquid contact medium. In another aspect of this invention, the physical and chemical performance of a direct contact condenser can be predicted based on the vapor and coolant compositions, the condensation conditions, and the geometric properties of the contact medium. 39 figs.

Method and apparatus for high-efficiency direct contact condensation

DOEpatents

Bharathan, Desikan; Parent, Yves; Hassani, A. Vahab

1999-01-01

A direct contact condenser having a downward vapor flow chamber and an upward vapor flow chamber, wherein each of the vapor flow chambers includes a plurality of cooling liquid supplying pipes and a vapor-liquid contact medium disposed thereunder to facilitate contact and direct heat exchange between the vapor and cooling liquid. The contact medium includes a plurality of sheets arranged to form vertical interleaved channels or passageways for the vapor and cooling liquid streams. The upward vapor flow chamber also includes a second set of cooling liquid supplying pipes disposed beneath the vapor-liquid contact medium which operate intermittently in response to a pressure differential within the upward vapor flow chamber. The condenser further includes separate wells for collecting condensate and cooling liquid from each of the vapor flow chambers. In alternate embodiments, the condenser includes a cross-current flow chamber and an upward flow chamber, a plurality of upward flow chambers, or a single upward flow chamber. The method of use of the direct contact condenser of this invention includes passing a vapor stream sequentially through the downward and upward vapor flow chambers, where the vapor is condensed as a result of heat exchange with the cooling liquid in the contact medium. The concentration of noncondensable gases in the resulting condensate-liquid mixtures can be minimized by controlling the partial pressure of the vapor, which depends in part upon the geometry of the vapor-liquid contact medium. In another aspect of this invention, the physical and chemical performance of a direct contact condenser can be predicted based on the vapor and coolant compositions, the condensation conditions. and the geometric properties of the contact medium.

An invisible medium for circularly polarized electromagnetic waves.

PubMed

Tamayama, Y; Nakanishi, T; Sugiyama, K; Kitano, M

2008-12-08

We study the no reflection condition for a planar boundary between vacuum and an isotropic chiral medium. In general chiral media, elliptically polarized waves incident at a particular angle satisfy the no reflection condition. When the wave impedance and wavenumber of the chiral medium are equal to the corresponding parameters of vacuum, one of the circularly polarized waves is transmitted to the medium without reflection or refraction for all angles of incidence. We propose a circular polarizing beam splitter as a simple application of the no reflection effect. (c) 2008 Optical Society of America

Establishment of a primary hepatocyte culture from the small Indian mongoose (Herpestes auropunctatus) and distribution of mercury in liver tissue.

PubMed

Horai, Sawako; Yanagi, Kumiko; Kaname, Tadashi; Yamamoto, Masatatsu; Watanabe, Izumi; Ogura, Go; Abe, Shintaro; Tanabe, Shinsuke; Furukawa, Tatsuhiko

2014-11-01

The present study established a primary hepatocyte culture for the small Indian mongoose (Herpestes auropunctatus). To determine the suitable medium for growing the primary hepatic cells of this species, we compared the condition of cells cultured in three media that are frequently used for mammalian cell culture: Dulbecco's Modified Eagle's Medium, RPMI-1640, and William's E. Of these, William's E medium was best suited for culturing the hepatic cells of this species. Using periodic acid-Schiff staining and ultrastructural observations, we demonstrated the cells collected from mongoose livers were hepatocytes. To evaluate the distribution of mercury (Hg) in the liver tissue, we carried out autometallography staining. Most of the Hg compounds were found in the central region of hepatic lobules. Smooth endoplasmic reticulum, which plays a role inxenobiotic metabolism, lipid/cholesterol metabolism, and the digestion and detoxification of lipophilic substances is grown in this area. This suggested that Hg colocalized with smooth endoplasmic reticulum. The results of the present study could be useful to identify the detoxification systems of wildlife with high Hg content in the body, and to evaluate the susceptibility of wildlife to Hg toxicity.

Proposal for a method to estimate nutrient shock effects in bacteria

PubMed Central

2012-01-01

Background Plating methods are still the golden standard in microbiology; however, some studies have shown that these techniques can underestimate the microbial concentrations and diversity. A nutrient shock is one of the mechanisms proposed to explain this phenomenon. In this study, a tentative method to assess nutrient shock effects was tested. Findings To estimate the extent of nutrient shock effects, two strains isolated from tap water (Sphingomonas capsulata and Methylobacterium sp.) and two culture collection strains (E. coli CECT 434 and Pseudomonas fluorescens ATCC 13525) were exposed both to low and high nutrient conditions for different times and then placed in low nutrient medium (R2A) and rich nutrient medium (TSA). The average improvement (A.I.) of recovery between R2A and TSA for the different times was calculated to more simply assess the difference obtained in culturability between each medium. As expected, A.I. was higher when cells were plated after the exposition to water than when they were recovered from high-nutrient medium showing the existence of a nutrient shock for the diverse bacteria used. S. capsulata was the species most affected by this phenomenon. Conclusions This work provides a method to consistently determine the extent of nutrient shock effects on different microorganisms and hence quantify the ability of each species to deal with sudden increases in substrate concentration. PMID:22873690

Human periodontal ligament stem cells secretome from multiple sclerosis patients suppresses NALP3 inflammasome activation in experimental autoimmune encephalomyelitis

PubMed Central

Soundara Rajan, Thangavelu; Giacoppo, Sabrina; Diomede, Francesca; Bramanti, Placido; Trubiani, Oriana; Mazzon, Emanuela

2017-01-01

Research in recent years has largely explored the immunomodulatory effects of mesenchymal stem cells (MSCs) and their secretory products, called “secretome,” in the treatment of neuroinflammatory diseases. Here, we examined whether such immunosuppressive effects might be elicited due to inflammasome inactivation. To this end, we treated experimental autoimmune encephalomyelitis (EAE) mice model of multiple sclerosis (MS) with the conditioned medium or purified exosomes/microvesicles (EMVs) obtained from relapsing-remitting-MS patients human periodontal ligament stem cells (hPDLSCs) and investigated the regulation of NALP3 inflammasome. We noticed enhanced expression of NALP3, Cleaved Caspase 1, interleukin (IL)-1β, and IL-18 in EAE mouse spinal cord. Conversely, hPDLSCs-conditioned medium and EMVs significantly blocked NALP3 inflammasome activation and provided protection from EAE. Reduction in NALP3, Cleaved Caspase 1, IL-1β, and IL-18 level was noticed in conditioned medium and EMVs-treated EAE mice. Pro-inflammatory Toll-like receptor (TLR)-4 and nuclear factor (NF)-κB were elevated in EAE, while hPDLSCs-conditioned medium and EMVs treatment reduced their expression and increased IκB-α expression. Characterization of hPDLSCs-conditioned medium showed substantial level of anti-inflammatory IL-10, transforming growth factor (TGF)-β, and stromal cell–derived factor 1α (SDF-1α). We propose that the immunosuppressive role of hPDLSCs-derived conditioned medium and EMVs in EAE mice may partly attribute to the presence of soluble immunomodulatory factors, NALP3 inflammasome inactivation, and NF-κB reduction. PMID:28764573

Encapsulated microsensors for reservoir interrogation

DOEpatents

Scott, Eddie Elmer; Aines, Roger D.; Spadaccini, Christopher M.

2016-03-08

In one general embodiment, a system includes at least one microsensor configured to detect one or more conditions of a fluidic medium of a reservoir; and a receptacle, wherein the receptacle encapsulates the at least one microsensor. In another general embodiment, a method include injecting the encapsulated at least one microsensor as recited above into a fluidic medium of a reservoir; and detecting one or more conditions of the fluidic medium of the reservoir.

Biological tracer method

DOEpatents

Strong-Gunderson, Janet M.; Palumbo, Anthony V.

1998-01-01

The present invention is a biological tracer method for characterizing the movement of a material through a medium, comprising the steps of: introducing a biological tracer comprising a microorganism having ice nucleating activity into a medium; collecting at least one sample of the medium from a point removed from the introduction point; and analyzing the sample for the presence of the biological tracer. The present invention is also a method for using a biological tracer as a label for material identification by introducing a biological tracer having ice nucleating activity into a material, collecting a sample of a portion of the labelled material and analyzing the sample for the presence of the biological tracer.

Biological tracer method

DOEpatents

Strong-Gunderson, J.M.; Palumbo, A.V.

1998-09-15

The present invention is a biological tracer method for characterizing the movement of a material through a medium, comprising the steps of: introducing a biological tracer comprising a microorganism having ice nucleating activity into a medium; collecting at least one sample of the medium from a point removed from the introduction point; and analyzing the sample for the presence of the biological tracer. The present invention is also a method for using a biological tracer as a label for material identification by introducing a biological tracer having ice nucleating activity into a material, collecting a sample of a portion of the labelled material and analyzing the sample for the presence of the biological tracer. 2 figs.

Optimization of culture conditions and medium composition for the production of micrococcin GO5 by Micrococcus sp. GO5.

PubMed

Kim, Mi-Hee; Kong, Yoon-Jung; Baek, Hong; Hyun, Hyung-Hwan

2006-01-02

To enhance the production of micrococcin GO5, a bacteriocin produced by Micrococcus sp. GO5, cultivation conditions and medium composition were optimized. The optimal initial pH and temperature for bacteriocin production were 7.0-9.0 and 37 degrees C, respectively. Micrococcus sp. GO5 displayed the highest micrococcin GO5 activity when grown in modified MRS medium that contained lactose or sucrose, rather than glucose, as a carbon source. The maximum bacteriocin activity was obtained in modified MRS medium containing 0.5% tryptone and 1.0% yeast extract as nitrogen sources instead of the other nitrogen sources present in MRS medium. Bacteriocin production was greatly affected by the concentration of K(2)HPO(4); strain GO5 produced eight-fold more bacteriocin in medium containing 2.0-2.5% K(2)HPO(4) than in medium containing 0.2% K(2)HPO(4). The optimal concentration of MgSO(4).7H(2)O for bacteriocin production was 0.5%. The production of micrococcin GO5 was increased 32-fold in shake flask culture and 16-fold in a bioreactor using the optimized medium (TY medium), compared with culturing in MRS medium.

MicroRNA-3178 ameliorates inflammation and gastric carcinogenesis promoted by Helicobacter pylori new toxin, Tip-α, by targeting TRAF3.

PubMed

Zou, Meijuan; Wang, Fang; Jiang, Aiqin; Xia, Anliang; Kong, Siya; Gong, Chun; Zhu, Mingxia; Zhou, Xin; Zhu, Jun; Zhu, Wei; Cheng, Wenfang

2017-04-01

Helicobacter pylori infection is the main cause of chronic gastritis, peptic ulcer, and gastric cancer. Tip-α is a newly identified carcinogenic factor present in H. pylori. TRAF3 can activate NF-κB by both canonical and noncanonical signaling pathways. In this study, we found that the expression of TRAF3 and NF-κB was upregulated, while microRNA-3178 (miR-3178) was decreased in H. pylori-positive gastric tissues but not in H. pylori-negative tissues. GES-1 cells were incubated with 12.5 μg/mL recombinant Tip-α (rTip-α) in RPMI1640 for 2 hours. After another 24 hours, the supernatant medium was designed as inflammatory-conditioned medium (ICM) and that from the untreated control cells was designed as untreated control medium. The release of proinflammatory cytokines from GES-1 cells and proliferation of gastric cancer cells was determined by ELISA and CCK-8 kits. Cells were transfected with the mimic, inhibitor, negative control of miR-3178, or TRAF3 siRNA control siRNA. The medium was then replaced with RPMI1640, 12.5 μg/mL rTip-α, and collected, and the total cellular RNA and protein were extracted for the following detection. MiR-3178 mimic prevented the increasement of TRAF3 and hence decreased activation of NF-κB signals, whereas miR-3178 inhibitor could not, in GES-1 cells with Tip-α treatment. The condition medium from miR-3178 mimic transfected GES-1 cells could inhibit proliferation and induce apoptosis of inflammation-related gastric cancer cells SGC7901 and MGC803 by decreasing the production of inflammatory cytokines TNF-α and IL-6, which were secreted by GES-1 cells. Taken all together, Tip-α might activate NF-κB to promote inflammation and carcinogenesis by inhibiting miR-3178 expression, which directly targeting TRAF3, during H. pylori infection in gastric mucosal epithelial cells. © 2016 John Wiley & Sons Ltd.

A new medium for Caenorhabditis elegans toxicology and nanotoxicology studies designed to better reflect natural soil solution conditions.

PubMed

Tyne, William; Lofts, Stephen; Spurgeon, David J; Jurkschat, Kerstin; Svendsen, Claus

2013-08-01

A new toxicity test medium for Caenorhabditis elegans is presented. The test solution is designed to provide a better representation of natural soil pore water conditions than currently available test media. The medium has a composition that can readily be modified to allow for studies of the influences of a range of environmentally relevant parameters on nematode biology and toxicology. Tests conducted in the new medium confirmed that nematodes' reproduction was possible at a range of solution pH levels, offering the potential to conduct toxicity studies under a variety of conditions. A test to establish silver nanoparticle and dissolved silver nitrate toxicity, a study type not feasible in M9 or agar media due to precipitation and nanoparticle agglomeration, indicated lower silver nanoparticle (median effective concentration [EC50] of 6.5 mg Ag/L) than silver nitrate (EC50 0.28 mg Ag/L) toxicity. Characterization identified stable nanoparticle behavior in the new test medium. Copyright © 2013 SETAC.

Oxidative stress differentially impacts male and female bovine embryos depending on the culture medium and the stress condition.

PubMed

Dallemagne, Matthew; Ghys, Emmanuelle; De Schrevel, Catalina; Mwema, Ariane; De Troy, Delphine; Rasse, Catherine; Donnay, Isabelle

2018-09-01

Male and female embryos are known to differ for their metabolism and response to environmental factors very early in development. The present study aimed to evaluate the response to oxidative stress of male and female bovine embryos at the morula-blastocyst stages in terms of developmental rates, total cell number and apoptotic rates in two culture conditions. Embryos where cultured in a medium supplemented with either 5% fetal calf serum (FCS) or 4 mg/mL bovine serum albumin and a mixture of insulin, transferrin and selenium (BSA-ITS). Oxidative stress was applied at Day-5 post insemination (pi) by adding either AAPH or menadione to the culture medium, and blastocysts were analyzed at Day-7pi. The impact on development and blastocyst quality was dependent on the culture medium and the stress inducer but differed between male and female embryos. Male embryos resisted better to oxidative stress in FCS supplemented medium, no matter the stress inducer. Accordingly, the impact on blastocyst cell number tended to be higher in female blastocysts after stress induction with AAPH in FCS supplemented medium. On the other hand, in BSA-ITS supplemented medium, female embryos were more resistant to AAPH induced stress, while menadione had no impact on sex ratio. The weaker resistance of males to AAPH in this medium is in accordance with their trend to show a higher increase in apoptotic rates than females in this condition. In conclusion, this study shows that oxidative stress has differential impact on male and female bovine blastocysts depending on the culture condition and on the way oxidative stress is induced. Copyright © 2018 Elsevier Inc. All rights reserved.

Technical note: Discard the specimen collection swab directly at point of care to improve extensive automated processing in clinical microbiology laboratories.

PubMed

Avolio, Manuela; Grosso, Shamanta; Bruschetta, Graziano; Camporese, Alessandro

2016-10-01

We compared, in terms of microorganisms recovery, the discard of specimen collection swab, after swirling into its medium, directly at point of care, with its placing into the medium and vortexing on arrival in the laboratory. Our results show that these two procedures are overlapped in terms of bacterial recovery. Copyright © 2016 Elsevier B.V. All rights reserved.

Lemna paucicostata Hegelm. 6746

PubMed Central

Datko, Anne H.; Mudd, S. Harvey; Giovanelli, John

1980-01-01

Photoautotrophic and mixotrophic growth of Lemna paucicostata Hegelm. 6746 (formerly Lemna perpusilla Torr. 6746) was investigated to establish standardized conditions for biochemical studies. Optimal temperature for growth was 29 to 30 C. The medium used previously (Datko AH, Mudd SH, Giovanelli J 1977 J Biol Chem 252: 3436-3445) was modified by inclusion of NH4Cl, decreasing macronutrient and ethylenediamine tetraacetate concentration, increasing micronutrient concentration, and inclusion of bicarbonate (for photoautotrophic growth) or 2-(N-morpholino)ethanesulfonic acid (for mixotrophic growth) buffers. Varying the sulfate concentration between 14 and 1 millimolar had no effect on growth. For photoautotrophic growth in the new medium (medium 4), the effects of CO2 concentration, light intensity, and pH were measured. Under the optimal conditions, a multiplication rate (MR) of 300 to 315, equivalent to a doubling time of 23 to 24 hours was obtained. Addition of glutamine or asparagine did not increase this MR. For mixotrophic growth in low light, the effects of sucrose concentration and pH were determined. Under optimal conditions, MR was 210. A concentration of sucrose less than maximal for growth was chosen for the medium for experiments which will include 14C-labeling of intermediates. MR under these conditions was 184. Growth was equally good in medium 4 and in half-strength Hutner's medium when sulfate was high (0.4 to 1 millimolar), but better in medium 4 when sulfate was low (20 micromolar). Growth rates could be restored to normal in half-strength Hutner's with low sulfate by decreasing the molybdate concentration. By modifying medium 4 to contain very low amounts of sulfate, and by preconditioning medium and plants, it was shown that there was an increment in plant protein of approximately 2.5 micrograms per nanomole of added MgSO4. Colonies undergoing sulfur limitation exhibited a slow growth rate and a high frond to colony ratio. Molybdate and selenate produced growth inhibition reversible by sulfate. Conditions were developed in which the plants could be maintained indefinitely in the presence of either molybdate or selenate in altered metabolic steady-states with lowered growth rates and protein per frond. Images PMID:16661306

Current collection by high voltage anodes in near ionospheric conditions

NASA Technical Reports Server (NTRS)

Antoniades, John A.; Greaves, Rod G.; Boyd, D. A.; Ellis, R.

1990-01-01

The authors experimentally identified three distinct regimes with large differences in current collection in the presence of neutrals and weak magnetic fields. In magnetic field/anode voltage space the three regions are separated by very sharp transition boundaries. The authors performed a series of laboratory experiments to study the dependence of the region boundaries on several parameters, such as the ambient neutral density, plasma density, magnetic field strength, applied anode voltage, voltage pulsewidth, chamber material, chamber size and anode radius. The three observed regimes are: classical magnetic field limited collection; stable medium current toroidal discharge; and large scale, high current space glow discharge. There is as much as several orders of magnitude of difference in the amount of collected current upon any boundary crossing, particularly if one enters the space glow regime. They measured some of the properties of the plasma generated by the breakdown that is present in regimes II and III in the vicinity of the anode including the sheath modified electrostatic potential, I-V characteristics at high voltage as well as the local plasma density.

Identifying hydrological regime and eco-flow threshold of small and medium flood of the Xiaoqing River in Jinan city

NASA Astrophysics Data System (ADS)

Liu, Yang; Cao, Sheng-Le

2017-06-01

It was known that hydrological regime was the main influencing factor of river ecosystem, but the regime of different flow rates of urban rivers was poorly understood. We collected daily inflows at the Huangtai station of the Xiaoqing River from 1960 to 2014 and divided the data into three periods. Then we calculated hydrological parameters by the method of EFCs (Environmental Flow Components) and analyzed the tendency and change rates of each component respectively in the three periods. Combined with the ecological significance of environmental flow components, we identified the small and medium flood had the greatest impact on the river regime and ecosystem. And then we used the hydraulic parameters in the good ecosystem period as control conditions, to calculate the ecological threshold of the flow component under the current situation. This study could provide technical support for restoring and improving hydrological regime and ecological environment of the Xiaoqing River in Jinan city.

Predictive variables for the occurrence of early clinical mastitis in primiparous Holstein cows under field conditions in France.

PubMed Central

Barnouin, J; Chassagne, M

2001-01-01

Holstein heifers from 47 dairy herds in France were enrolled in a field study to determine predictors for clinical mastitis within the first month of lactation. Precalving and calving variables (biochemical, hematological, hygienic, and disease indicators) were collected. Early clinical mastitis (ECM) predictive variables were analyzed by using a multiple logistic regression model (99 cows with ECM vs. 571 without clinical mastitis throughout the first lactation). Two variables were associated with a higher risk of ECM: a) difficult calving and b) medium and high white blood cell (WBC) counts in late gestation. Two prepartum indicators were associated with a lower ECM risk: a) medium and high serum concentrations of immunoglobulin G1 (IgG1) and b) high percentage of eosinophils among white blood cells. Calving difficulty and certain biological blood parameters (IgG1, eosinophils) could represent predictors that would merit further experimental studies, with the aim of designing programs for reducing the risk of clinical mastitis in the first lactation. PMID:11195522

Stimulation of GABA-Induced Ca2+ Influx Enhances Maturation of Human Induced Pluripotent Stem Cell-Derived Neurons

PubMed Central

Rushton, David J.; Mattis, Virginia B.; Svendsen, Clive N.; Allen, Nicholas D.; Kemp, Paul J.

2013-01-01

Optimal use of patient-derived, induced pluripotent stem cells for modeling neuronal diseases is crucially dependent upon the proper physiological maturation of derived neurons. As a strategy to develop defined differentiation protocols that optimize electrophysiological function, we investigated the role of Ca2+ channel regulation by astrocyte conditioned medium in neuronal maturation, using whole-cell patch clamp and Ca2+ imaging. Standard control medium supported basic differentiation of induced pluripotent stem cell-derived neurons, as assayed by the ability to fire simple, single, induced action potentials. In contrast, treatment with astrocyte conditioned medium elicited complex and spontaneous neuronal activity, often with rhythmic and biphasic characteristics. Such augmented spontaneous activity correlated with astrocyte conditioned medium-evoked hyperpolarization and was dependent upon regulated function of L-, N- and R-type Ca2+ channels. The requirement for astrocyte conditioned medium could be substituted by simply supplementing control differentiation medium with high Ca2+ or γ-amino butyric acid (GABA). Importantly, even in the absence of GABA signalling, opening Ca2+ channels directly using Bay K8644 was able to hyperpolarise neurons and enhance excitability, producing fully functional neurons. These data provide mechanistic insight into how secreted astrocyte factors control differentiation and, importantly, suggest that pharmacological modulation of Ca2+ channel function leads to the development of a defined protocol for improved maturation of induced pluripotent stem cell-derived neurons. PMID:24278369

Neutrino flavor instabilities in a time-dependent supernova model

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

Abbar, Sajad; Duan, Huaiyu

2015-10-19

In this study, a dense neutrino medium such as that inside a core-collapse supernova can experience collective flavor conversion or oscillations because of the neutral-current weak interaction among the neutrinos. This phenomenon has been studied in a restricted, stationary supernova model which possesses the (spatial) spherical symmetry about the center of the supernova and the (directional) axial symmetry around the radial direction. Recently it has been shown that these spatial and directional symmetries can be broken spontaneously by collective neutrino oscillations. In this letter we analyze the neutrino flavor instabilities in a time-dependent supernova model. Our results show that collectivemore » neutrino oscillations start at approximately the same radius in both the stationary and time-dependent supernova models unless there exist very rapid variations in local physical conditions on timescales of a few microseconds or shorter. Our results also suggest that collective neutrino oscillations can vary rapidly with time in the regimes where they do occur which need to be studied in time-dependent supernova models.« less

Measurements of bovine sperm velocities under true anaerobic and aerobic conditions.

PubMed

Krzyzosiak, J; Molan, P; Vishwanath, R

1999-04-30

Velocities of bovine spermatozoa in a medium containing glucose were similar under true anaerobic and aerobic conditions. Spermatozoa were not able to sustain motility under anaerobic conditions when glycolysis was inhibited, but regained motility when re-aerated. This demonstrates that immobilisation was due to lack of oxygen and that conditions under which motility was analysed were truly anaerobic. Sperm motility parameters were not significantly different in the presence and absence of 4 microM antimycin A and 4 microM rotenone when glucose was present in the medium. After each incubation, functionality of sperm mitochondria was assayed by washing sperm into the medium which supported respiration but not glycolysis, and motility was visually assessed. All sperm samples were highly motile in this medium indicating that their mitochondria were functional. When glycolysis was inhibited, antimycin and rotenone abolished sperm motility immediately after addition. Bovine sperm can maintain similar levels of motility aerobically and anaerobically if a glycolysable substrate is available. Available data on bovine sperm energetics support this view.

Role for RIP1 in mediating necroptosis in experimental intracerebral hemorrhage model both in vivo and in vitro.

PubMed

Shen, Haitao; Liu, Chenglin; Zhang, Dongping; Yao, Xiyang; Zhang, Kai; Li, Haiying; Chen, Gang

2017-03-02

Cell death is a hallmark of second brain injury after intracerebral hemorrhage (ICH); however, the mechanism still has not been fully illustrated. In this study, we explored whether necroptosis, a type of regulated necrosis, has an essential role in brain injury after ICH. We found that inhibiting receptor-interacting protein 1 (RIP1) - a core element of the necroptotic pathway - by a specific chemical inhibitor or genetic knockdown attenuated brain injury in a rat model of ICH. Furthermore, necroptosis of cultured neurons could be induced by conditioned medium from microglia stimulated with oxygen hemoglobin, and this effect could be inhibited by TNF-α inhibitor, indicating that TNF-α secreted from activated microglia is an important factor in inducing necroptosis of neurons. Undoubtedly, overexpression of RIP1 increased conditioned medium-induced necroptosis in vitro, but this effect was partially diminished in mutation of serine kinase phosphorylation site of RIP1, showing that phosphorylation of RIP1 is the essential molecular mechanism of necroptosis, which was activated in the in vitro model of ICH. Collectively, our investigation identified that necroptosis is an important mechanism of cell death in brain injury after ICH, and inhibition of necroptosis may be a potential therapeutic intervention of ICH.

Role for RIP1 in mediating necroptosis in experimental intracerebral hemorrhage model both in vivo and in vitro

PubMed Central

Shen, Haitao; Liu, Chenglin; Zhang, Dongping; Yao, Xiyang; Zhang, Kai; Li, Haiying; Chen, Gang

2017-01-01

Cell death is a hallmark of second brain injury after intracerebral hemorrhage (ICH); however, the mechanism still has not been fully illustrated. In this study, we explored whether necroptosis, a type of regulated necrosis, has an essential role in brain injury after ICH. We found that inhibiting receptor-interacting protein 1 (RIP1) – a core element of the necroptotic pathway – by a specific chemical inhibitor or genetic knockdown attenuated brain injury in a rat model of ICH. Furthermore, necroptosis of cultured neurons could be induced by conditioned medium from microglia stimulated with oxygen hemoglobin, and this effect could be inhibited by TNF-α inhibitor, indicating that TNF-α secreted from activated microglia is an important factor in inducing necroptosis of neurons. Undoubtedly, overexpression of RIP1 increased conditioned medium-induced necroptosis in vitro, but this effect was partially diminished in mutation of serine kinase phosphorylation site of RIP1, showing that phosphorylation of RIP1 is the essential molecular mechanism of necroptosis, which was activated in the in vitro model of ICH. Collectively, our investigation identified that necroptosis is an important mechanism of cell death in brain injury after ICH, and inhibition of necroptosis may be a potential therapeutic intervention of ICH. PMID:28252651

Evaluation of the interferon-γ assay on blood collected at exsanguination of cattle under field conditions for surveillance of bovine tuberculosis.

PubMed

Okafor, C C; Grooms, D L; Bolin, S R; Averill, J J; Kaneene, J B

2014-12-01

Development of point of concentration (POC) surveillance strategies for bovine tuberculosis (bTB) would facilitate global efforts to eradicate bTB. The interferon-gamma (IFNγ) assay can detect IFNγ responses to Mycobacterium bovis in blood collected at commencement of exsanguination (COE) of experimentally challenged cattle but has not been evaluated under field conditions. The current study was aimed at determining (i) whether blood collected at COE of cattle at slaughter, under field conditions, is practical to obtain and useful for identifying cattle as IFNγ positive for bTB, (ii) whether the results of the IFNγ assay obtained at COE reliably compare with results obtained from live animals in the field, and (iii) whether the identified animal(s) originated from bTB-infected or bTB-exposed herds. Cattle from three risk groups were used: the highest risk group consisted of 49 cattle from 3 bTB-infected herds; the medium risk group consisted of 24 cattle from a potentially exposed herd; and the lowest risk group consisted of 60 cattle from herds with no known history of bTB exposure. The IFNγ assay was performed on blood collected both before stunning and at COE of cattle at slaughter. An enhanced slaughter inspection for gross lesions consistent with bTB was performed on all cattle. In addition, lymph nodes were cultured for M. bovis for cattle that tested positive for bTB via the IFNγ assay and for most cattle that tested negative for bTB. Cattle, both with and without lesions consistent with bTB, were identified as positive for bTB by the IFNγ assay using blood collected at COE, but none of the positive cattle originated from the lowest risk group. The current study demonstrates that blood collected at COE of cattle is both a practical and moderately reliable sample for accessing bTB infection using the IFNγ assay. © 2013 Blackwell Verlag GmbH.

Conditioned medium as a strategy for human stem cells chondrogenic differentiation.

PubMed

Alves da Silva, M L; Costa-Pinto, A R; Martins, A; Correlo, V M; Sol, P; Bhattacharya, M; Faria, S; Reis, R L; Neves, Nuno M

2015-06-01

Paracrine signalling from chondrocytes has been reported to increase the synthesis and expression of cartilage extracellular matrix (ECM) by stem cells. The use of conditioned medium obtained from chondrocytes for stimulating stem cells chondrogenic differentiation may be a very interesting alternative for moving into the clinical application of these cells, as chondrocytes could be partially replaced by stem cells for this type of application. In the present study we aimed to achieve chondrogenic differentiation of two different sources of stem cells using conditioned medium, without adding growth factors. We tested both human bone marrow-derived mesenchymal stem cells (hBSMCs) and human Wharton's jelly-derived stem cells (hWJSCs). Conditioned medium obtained from a culture of human articular chondrocytes was used to feed the cells during the experiment. Cultures were performed in previously produced three-dimensional (3D) scaffolds, composed of a blend of 50:50 chitosan:poly(butylene succinate). Both types of stem cells were able to undergo chondrogenic differentiation without the addition of growth factors. Cultures using hWJSCs showed significantly higher GAGs accumulation and expression of cartilage-related genes (aggrecan, Sox9 and collagen type II) when compared to hBMSCs cultures. Conditioned medium obtained from articular chondrocytes induced the chondrogenic differentiation of MSCs and ECM formation. Obtained results showed that this new strategy is very interesting and should be further explored for clinical applications. Copyright © 2013 John Wiley & Sons, Ltd.

[Occupational and non-occupational factors influencing health state of small and medium business workers].

PubMed

Fasikov, R M; Khuzhakhmetova, I B; Stepanov, E G

2010-01-01

Complex study of work conditions and health parameters of workers engaged into small and medium business proved that preserved and better health of these workers, prevention of occupational and occupationally mediated diseases necessitate federal and regional complex system of measures including legal basis, database on work conditions and their influence on small and medium business workers' health, occupational medicine training for employers and employees, more active involvement of medical institutioins into screening for occupational diseases.

Analysis of successful rate factors for small and medium enterprises in furniture manufacturing sector in Klaten Regency - Central Java, Indonesia

NASA Astrophysics Data System (ADS)

Budhi Utomo, R.; Lasminiasih; Prajaka, S.

2018-03-01

Small and Medium Enterprises (SMEs) are business activities that can expand the level of employment rate and provide economic services to the wider community and can play a role in the process of equalizing and improving people’s income, stimulating economic growth as well as realizing national stabilities. The aim of this study is to identify the factors of the success rate for Small and Medium Enterprises (SMEs) in furniture manufacturing sector in Klaten regency, Central Java, Indonesia. The method employed in this study was descriptive qualitative by also employing quantitative analysis of which the data were collected through observations, interviews and by administering questionnaires. The results seemed to indicate that the furniture business in Klaten is still experiencing difficulties in managing its various aspects of business, namely in terms of marketing (either directly or indirectly or by making the best use media of technology) and managing capital. All this time, the SMEs in furniture manufacturing sector in Klaten have been utilizing a very simple system in producing tables, chairs, wardrobes and any other furniture products which are then distributed to be sold by larger furniture companies. This condition makes the SMEs unable to be independent in running their business.

Comparison of Vehicle-Broadcasted Fuel Consumption Rates against Precise Fuel Measurements for Medium- and Heavy-Duty Vehicles and Engines

DOE PAGES

Pink, Alex; Ragatz, Adam; Wang, Lijuan; ...

2017-03-28

Vehicles continuously report real-time fuel consumption estimates over their data bus, known as the controller area network (CAN). However, the accuracy of these fueling estimates is uncertain to researchers who collect these data from any given vehicle. To assess the accuracy of these estimates, CAN-reported fuel consumption data are compared against fuel measurements from precise instrumentation. The data analyzed consisted of eight medium/heavy-duty vehicles and two medium-duty engines. Varying discrepancies between CAN fueling rates and the more accurate measurements emerged but without a vehicular trend-for some vehicles the CAN under-reported fuel consumption and for others the CAN over-reported fuel consumption.more » Furthermore, a qualitative real-time analysis revealed that the operating conditions under which these fueling discrepancies arose varied among vehicles. A drive cycle analysis revealed that while CAN fueling estimate accuracy differs for individual vehicles, that CAN estimates capture the relative fuel consumption differences between drive cycles within 4% for all vehicles and even more accurately for some vehicles. Furthermore, in situations where only CAN-reported data are available, CAN fueling estimates can provide relative fuel consumption trends but not accurate or precise fuel consumption rates.« less

Comparison of Vehicle-Broadcasted Fuel Consumption Rates against Precise Fuel Measurements for Medium- and Heavy-Duty Vehicles and Engines

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

Pink, Alex; Ragatz, Adam; Wang, Lijuan

Vehicles continuously report real-time fuel consumption estimates over their data bus, known as the controller area network (CAN). However, the accuracy of these fueling estimates is uncertain to researchers who collect these data from any given vehicle. To assess the accuracy of these estimates, CAN-reported fuel consumption data are compared against fuel measurements from precise instrumentation. The data analyzed consisted of eight medium/heavy-duty vehicles and two medium-duty engines. Varying discrepancies between CAN fueling rates and the more accurate measurements emerged but without a vehicular trend-for some vehicles the CAN under-reported fuel consumption and for others the CAN over-reported fuel consumption.more » Furthermore, a qualitative real-time analysis revealed that the operating conditions under which these fueling discrepancies arose varied among vehicles. A drive cycle analysis revealed that while CAN fueling estimate accuracy differs for individual vehicles, that CAN estimates capture the relative fuel consumption differences between drive cycles within 4% for all vehicles and even more accurately for some vehicles. Furthermore, in situations where only CAN-reported data are available, CAN fueling estimates can provide relative fuel consumption trends but not accurate or precise fuel consumption rates.« less

40 CFR Table 1 to Subpart Hhh of... - Emission Limits for Small Rural, Small, Medium, and Large HMIWI

Code of Federal Regulations, 2011 CFR

2011-07-01

..., Medium, and Large HMIWI 1 Table 1 to Subpart HHH of Part 62 Protection of Environment ENVIRONMENTAL... 62—Emission Limits for Small Rural, Small, Medium, and Large HMIWI Pollutant Units (7 percent oxygen, dry basis at standard conditions) Emission limits HMIWI size Small rural Small Medium Large...

40 CFR Table 1 to Subpart Hhh of... - Emission Limits for Small Rural, Small, Medium, and Large HMIWI

Code of Federal Regulations, 2012 CFR

2012-07-01

..., Medium, and Large HMIWI 1 Table 1 to Subpart HHH of Part 62 Protection of Environment ENVIRONMENTAL... 62—Emission Limits for Small Rural, Small, Medium, and Large HMIWI Pollutant Units (7 percent oxygen, dry basis at standard conditions) Emission limits HMIWI size Small rural Small Medium Large...

40 CFR Table 1 to Subpart Hhh of... - Emission Limits for Small Rural, Small, Medium, and Large HMIWI

Code of Federal Regulations, 2010 CFR

2010-07-01

..., Medium, and Large HMIWI 1 Table 1 to Subpart HHH of Part 62 Protection of Environment ENVIRONMENTAL... 62—Emission Limits for Small Rural, Small, Medium, and Large HMIWI Pollutant Units (7 percent oxygen, dry basis at standard conditions) Emission limits HMIWI size Small rural Small Medium Large...

Multiple shoot induction from axillary bud cultures of the medicinal orchid, Dendrobium longicornu

PubMed Central

Dohling, Stadwelson; Kumaria, Suman; Tandon, Pramod

2012-01-01

Background and aims Dendrobium longicornu, commonly known as the ‘Long-horned Dendrobium’, is an endangered and medicinally important epiphytic orchid. Over-exploitation and habitat destruction seriously threaten this orchid in Northeast India. Our objective was to develop an efficient protocol for the mass propagation of D. longicornu using axillary bud segments. Methodology and principal results Axillary buds cultured in Murashige and Skoog semi-solid medium supplemented with α-naphthalene acetic acid (NAA), 2,4-dichlorophenoxy acetic acid (2,4-D) and 6-benzylaminopurine (BAP) readily developed into plantlets. These formed either directly from shoot buds or from intermediary protocorm-like bodies (PLBs). The maximum explant response (86.6 %) was obtained in medium supplemented with NAA at 30 µM, while the maximum number of shoots (4.42) and maximum bud-forming capacity (3.51) were observed in medium containing 15 µM BAP and 5 µM NAA in combination. Protocorm-like bodies were obtained when the medium contained 2,4-D. The maximum number of explants forming PLBs (41.48 %) was obtained in medium containing 15 µM BAP and 15 µM 2,4-D. Well-developed plantlets obtained after 20–25 weeks of culture were acclimatized and eventually transferred to the greenhouse. Over 60 % of these survived to form plants ∼3–4 cm tall after 90 days in glasshouse conditions using a substrate of crushed brick and charcoal, shredded bark and moss. Conclusions The method described can readily be used for the rapid and large-scale regeneration of D. longicornu. Its commercial adoption would reduce the collection of this medicinally important and increasingly rare orchid from the wild. PMID:23136638

Callus induction and flavonoid production on the immature seed of Stelechocarpus burahol

NASA Astrophysics Data System (ADS)

Habibah, N. A.; Moeljopawiro, S.; Dewi, K.; Indrianto, A.

2018-03-01

Stelechocarpus burahol [(Bl.) Hook. f. & Th.] is one of the medicinal plants. In vitro callus induction studies on S. burahol were carried out to determine phytohormone requirement for optimum callus induction. Immature seed explants were cultured on MS medium by adding different kinds and different concentrations of plant growth regulators (picloram and 2,4-D) under light and dark conditions. The results showed that callus formation was initiated on the 18,50th to the 55th days. The best condition for optimum callus induction was found on MS medium, which was supplemented with 7.5 mg/L picloram and was maintained in the dark condition. The callus induction varied from 60% to 100%. The callus that produced the highest flavonoid was grown on the medium with the addition of 10 mg/L of 2,4-D. In conclusion, the results represented a suitable medium for S.burahol callus induction.

Characterization of In-Use Medium Duty Electric Vehicle Driving and Charging Behavior: Preprint

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

Duran, A.; Ragatz, A.; Prohaska, R.

2014-11-01

The U.S. Department of Energy's American Recovery and Reinvestment Act (ARRA) deployment and demonstration projects are helping to commercialize technologies for all-electric vehicles (EVs). Under the ARRA program, data from Smith Electric and Navistar medium duty EVs have been collected, compiled, and analyzed in an effort to quantify the impacts of these new technologies. Over a period of three years, the National Renewable Energy Laboratory (NREL) has compiled data from over 250 Smith Newton EVs for a total of over 100,000 days of in-use operation. Similarly, data have been collected from over 100 Navistar eStar vehicles, with over 15,000 operatingmore » days having been analyzed. NREL has analyzed a combined total of over 4 million kilometers of driving and 1 million hours of charging data for commercial operating medium duty EVs. In this paper, the authors present an overview of medium duty EV operating and charging behavior based on in-use data collected from both Smith and Navistar vehicles operating in the United States. Specifically, this paper provides an introduction to the specifications and configurations of the vehicles examined; discusses the approach and methodology of data collection and analysis, and presents detailed results regarding daily driving and charging behavior. In addition, trends observed over the course of multiple years of data collection are examined, and conclusions are drawn about early deployment behavior and ongoing adjustments due to new and improving technology. Results and metrics such as average daily driving distance, route aggressiveness, charging frequency, and liter per kilometer diesel equivalent fuel consumption are documented and discussed.« less

Influence of Post-Mortem Sperm Recovery Method and Extender on Unstored and Refrigerated Rooster Sperm Variables.

PubMed

Villaverde-Morcillo, S; Esteso, M C; Castaño, C; Santiago-Moreno, J

2016-02-01

Many post-mortem sperm collection techniques have been described for mammalian species, but their use in birds is scarce. This paper compares the efficacy of two post-mortem sperm retrieval techniques - the flushing and float-out methods - in the collection of rooster sperm, in conjunction with the use of two extenders, i.e., L&R-84 medium and Lake 7.1 medium. To determine whether the protective effects of these extenders against refrigeration are different for post-mortem and ejaculated sperm, pooled ejaculated samples (procured via the massage technique) were also diluted in the above extenders. Post-mortem and ejaculated sperm variables were assessed immediately at room temperature (0 h), and after refrigeration at 5°C for 24 and 48 h. The flushing method retrieved more sperm than the float-out method (596.5 ± 75.4 million sperm vs 341.0 ± 87.6 million sperm; p < 0.05); indeed, the number retrieved by the former method was similar to that obtained by massage-induced ejaculation (630.3 ± 78.2 million sperm). For sperm collected by all methods, the L&R-84 medium provided an advantage in terms of sperm motility variables at 0 h. In the refrigerated sperm samples, however, the Lake 7.1 medium was associated with higher percentages of viable sperm, and had a greater protective effect (p < 0.05) with respect to most motility variables. In conclusion, the flushing method is recommended for collecting sperm from dead birds. If this sperm needs to be refrigerated at 5°C until analysis, Lake 7.1 medium is recommended as an extender. © 2015 Blackwell Verlag GmbH.

Growth medium alterations improve in vitro cold storage of pear germplasm.

PubMed

Kovalchuk, I; Zhumagulova, Z; Turdiev, T; Reed, B M

2014-01-01

Development of new fruit cultivars is dependent on genetic resource collections such as those at the Pomological Garden of the Institute of Horticulture and Viticulture near Almaty, Kazakhstan. The pear germplasm collection of the Pomological Garden contains 615 cultivars and three species. In vitro cold storage of the collection would provide additional security to the field collection. This study was designed to improve medium-term in vitro storage of pear germplasm. Shoots of seven pear cultivars (Pyrus communis L.) were stored in plastic five-section bags at 4 degree C and a 10-h photoperiod (7 μmol/m2/s). Treatments included medium with four carbohydrate sources (3% sucrose, 2% or 3% mannitol, or 2% sucrose + 2% mannitol) with 0.5 mg/l BAP and 0.1 mg/l IBA or without plant growth regulators (PGRs) and at three Murashige and Skoog (MS) nitrogen concentrations (100%, 50% or 25%). Pear shoots remained viable for 9 to 15 months without repropagation on the control MS medium with 3% sucrose without PGRs. There were significant impacts of cultivar and treatment on the duration of cold storage. Shoots of 'Mramornaya' remained viable (rating of ≥ 2) for 27 months with PGRs and 2% sucrose + 2% mannitol compared to 12 months for the PGR + 3% sucrose treatment. Talgarskaya Krasaviza stored for 18 months on 2% sucrose + 2% mannitol while all other treatments lasted only 6 to 9 months. Treatments with 0.5 or 1 mg/l abscisic acid (ABA) with 3% sucrose increased storage duration as did reducing the concentration of nitrogen in the medium to 25% without PGRs and with 3% sucrose.

Hydride Molecules towards Nearby Galaxies

NASA Astrophysics Data System (ADS)

Monje, Raquel R.; La, Ngoc; Goldsmith, Paul

2018-06-01

Observations carried out by the Herschel Space Observatory revealed strong spectroscopic signatures from light hydride molecules within the Milky Way and nearby active galaxies. To better understand the chemical and physical conditions of the interstellar medium, we conducted the first comprehensive survey of hydrogen fluoride (HF) and water molecular lines observed through the SPIRE Fourier Transform Spectrometer. By collecting and analyzing the sub-millimeter spectra of over two hundred sources, we found that the HF J = 1 - 0 rotational transition which occurs at approximately 1232 GHz was detected in a total of 39 nearby galaxies both in absorption and emission. The analysis will determine the main excitation mechanism of HF in nearby galaxies and provide steady templates of the chemistry and physical conditions of the ISM to be used in the early universe, where observations of hydrides are more scarce.

Remote sensing of Northern mines: supporting operation and environmental monitoring in cold conditions

NASA Astrophysics Data System (ADS)

Tuomela, Anne; Davids, Corine; Knutsson, Sven; Knutsson, Roger; Rauhala, Anssi; Rossi, Pekka M.; Rouyet, Line

2017-04-01

Northern areas of Finland, Sweden and Norway have mineral-rich deposits. There are several active mines in the area but also closed ones and deposits with plans for future mining. With increasing demand for environmental protection in the sensitive Northern conditions, there is a need for more comprehensive monitoring of the mining environment. In our study, we aim to develop new opportunities to use remote sensing data from satellites and unmanned aerial vehicles (UAVs) in improving mining safety and monitoring, for example in the case of mine waste storage facilities. Remote sensing methods have evolved fast, and could in many cases enable precise, reliable, and cost-efficient data collection over large areas. The study has focused on four mining areas in Northern Fennoscandia. Freely available medium-resolution (e.g. Sentinel-1), commercial high-resolution (e.g. TerraSAR-X) and Synthetic Aperture Radar (SAR) data has been collected during 2015-2016 to study how satellite remote sensing could be used e.g. for displacement monitoring using SAR Interferometry (InSAR). Furthermore, UAVs have been utilized in similar data collection in a local scale, and also in collection of thermal infrared data for hydrological monitoring of the areas. The development and efficient use of the methods in mining areas requires experts from several fields. In addition, the Northern conditions with four distinct seasons bring their own challenges for the efficient use of remote sensing, and further complicate their integration as standardised monitoring methods for mine environments. Based on the initial results, remote sensing could especially enhance the monitoring of large-scale structures in mine areas such as tailings impoundments.

32 CFR 310.15 - General considerations.

Code of Federal Regulations, 2014 CFR

2014-07-01

... individual's rights, privileges, or benefits under any Federal program. (b) Collecting social security... regardless of the medium used to collect the information (forms, personal interviews, telephonic interviews...

32 CFR 310.15 - General considerations.

Code of Federal Regulations, 2010 CFR

2010-07-01

... individual's rights, privileges, or benefits under any Federal program. (b) Collecting social security... regardless of the medium used to collect the information (forms, personal interviews, telephonic interviews...

How AACR2 Will Affect a Medium Sized Library.

ERIC Educational Resources Information Center

Pang, Isabel S.

1980-01-01

Measures the impact of AACR2 on the catalog of a medium sized college library, using data collected from the Library of Congress announced changes. How to deal with these changes and estimate their costs is discussed. (Author/RAA)

Development of conditioning programs for dressage horses based on time-motion analysis of competitions.

PubMed

Clayton, H M

1993-05-01

The time-motion characteristics of Canadian basic- and medium-level dressage competitions are described, and the results are applied in formulating sport-specific conditioning programs. One competition was analyzed at the six levels from basic 1 to medium 3. Each test was divided into a series of sequences based on the type and speed of activity. The durations of the sequences were measured from videotapes. The basic-level tests had fewer sequences, and they were shorter in distance and duration than the medium tests (P < 0.10), but the average speed did not differ between the two levels. It is recommended that horses competing at the basic levels be conditioned using 5-min exercise periods, with short (10-s) bursts of lengthened trot and canter included at basic 2 and above. In preparation for medium-level competitions, the duration of the work periods increases to 7 min, 10- to 12-s bursts of medium or extended trot and canter are included, and transitions are performed frequently to simulate the energy expenditure in overcoming inertia.

Mechanisms for the target patterns formation in a stochastic bistable excitable medium

NASA Astrophysics Data System (ADS)

Verisokin, Andrey Yu.; Verveyko, Darya V.; Postnov, Dmitry E.

2018-04-01

We study the features of formation and evolution of spatiotemporal chaotic regime generated by autonomous pacemakers in excitable deterministic and stochastic bistable active media using the example of the FitzHugh - Nagumo biological neuron model under discrete medium conditions. The following possible mechanisms for the formation of autonomous pacemakers have been studied: 1) a temporal external force applied to a small region of the medium, 2) geometry of the solution region (the medium contains regions with Dirichlet or Neumann boundaries). In our work we explore the conditions for the emergence of pacemakers inducing target patterns in a stochastic bistable excitable system and propose the algorithm for their analysis.

FIBER AND INTEGRATED OPTICS. OTHER TOPICS IN QUANTUM ELECTRONICS: Determination of the saturation parameter in a fast-flow CO2 active medium

NASA Astrophysics Data System (ADS)

Lebedev, V. F.; Lysikov, A. Yu; Ryazanova, A. V.; Shalygin, S. V.

1990-05-01

A diagnostic method was developed for determination of such an important characteristic of a CO2 active medium as the energy stored in the vibrational degrees of freedom of molecules, which can be determined directly from the experimentally measured powers of probe beams without the need for any additional information on the medium and without any numerical calculations. Measurements were carried out using a real fast-flow system, which satisfied the conditions of validity of the proposed diagnostic method, and the correctness of the approach was confirmed. The results obtained demonstrated the efficiency of the excitation of the medium when the pump conditions were varied.

A new method for evaluating the dissolution of orodispersible films.

PubMed

Xia, Yiran; Chen, Fang; Zhang, Huiping; Luo, Chunlin

2015-05-01

The aim of this research was to develop and assess a new dissolution apparatus for orodispersible films (ODFs). The new apparatus was based on a flow-through cell design which requires only a limited amount of dissolution medium and can automatically collect samples in short-time intervals. Compared with the dissolution method in Chinese Pharmacopeia, our method simulated the flow condition of the oral cavity and resulted in reproducible dissolution data and remarkably discriminating capability. Therefore, we concluded that the proposed dissolution method was particularly suitable for evaluating the dissolution of ODFs and should also be applicable to other fast-dissolving solid dosage forms.

Effects of neural stem cell media on hypoxic injury in rat hippocampal slice cultures.

PubMed

Lee, Na Mi; Chae, Soo Ahn; Lee, Hong Jun

2017-12-15

Neonatal hypoxic-ischemic brain injuries cause serious neurological sequelae, yet there is currently no effective treatment for them. We hypothesized that neurotrophic factors released into the medium by stem cells could supply hypoxia-damaged organotypic hippocampal slice cultures with regenerative abilities. We prepared organotypic slice cultures of the hippocampus of 7-day-old Sprague-Dawley rats based on the modified Stoppini method; slices were cultured for 14days in vitro using either Gahwiler's medium (G-medium) or stem cell-conditioned medium (S-medium) as culture medium. At day 14 in vitro, hippocampal slice cultures were exposed to 95% N 2 and 5% CO 2 for 3h to induce hypoxic damage, the extent of which was then measured using propidium iodide fluorescence and immunohistochemistry images. We performed dot blotting to estimate neurotrophic/growth factor levels in the G- and S-media. Organotypic hippocampal slices cultured using S-medium after hypoxic injury were significantly less damaged than those cultured using G-medium. GLUT1, NGF, GDNF, VEGF, GCSF, and IGF2 levels were higher in S-medium than in G-medium, whereas FGF1, HIF, and MCP3 levels were not significantly different between media. In conclusion, we found that stem cell-conditioned medium had a neuroprotective effect against hypoxic injury, and that, of the various neurotrophic factors in S-medium, NGF, GDNF, and VEGF can contribute to neuroprotection. Copyright © 2017 Elsevier B.V. All rights reserved.

[Method of culturing microorganisms at constant concentrations of the nutrient components].

PubMed

Markvichev, N S; Manakov, M N

1985-01-01

A method for batch cultivation of microorganisms in a flow medium is described, characterized by slight changes in concentrations of medium components in time and by the absence of products of vital activity of microorganisms in the fermentation medium. The conditions are achieved due to application of a fermentation installation with a microfiltrative membrane that separates the cells of cultivated microorganisms from the culture fluid and due to increasing the flow rate to a value at which the inlet and outlet concentrations of the medium components are almost equal. The cells of cultivated microorganisms under such conditions remain in the fermentation medium volume. The system was called "Ekostat". If the process is performed in "Ekostat" system, a positive deviation from the logarithmic law is observed for the growth rate of the yeast Candida utilis VSB-651 on ethanol cultivation.

CD109 is a component of exosome secreted from cultured cells

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

Sakakura, Hiroki; Department of Oral and Maxillofacial Surgery, Nagoya University Graduate School of Medicine, Nagoya; Mii, Shinji

Exosomes are 50–100-nm-diameter membrane vesicles released from various types of cells. Exosomes retain proteins, mRNAs and miRNAs, which can be transported to surrounding cells. CD109 is a glycosylphosphatidylinositol-anchored glycoprotein, and is released from the cell surface to the culture medium in vitro. Recently, it was reported that secreted CD109 from the cell surface downregulates transforming growth factor-β signaling in human keratinocytes. In this study, we revealed that CD109 is a component of the exosome in conditioned medium. FLAG-tagged human CD109 (FLAG-CD109) in conditioned medium secreted from HEK293 cells expressing FLAG-CD109 (293/FLAG-CD109) was immunoprecipitated with anti-FLAG affinity gel, and the co-precipitated proteins weremore » analyzed by mass spectrometry and western blotting. Exosomal proteins were associated with CD109. We revealed the presence of CD109 in exosome fractions from conditioned medium of 293/FLAG-CD109. Moreover, the localization of CD109 in the exosome was demonstrated using immuno-electron microscopy. When we used HEK293 cells expressing FLAG-tagged truncated CD109, which does not contain the C-terminal region, the association of truncated CD109 with exosomes was not detected in conditioned medium. These findings indicate that CD109 is an exosomal protein and that the C-terminal region of CD109 is required for its presence in the exosome. - Highlights: • CD109 is an exosomal protein. • The C-terminal region of CD109 is required for its presence in the exosome. • Part of the secreted CD109 is present in the exosome-free fraction in the conditioned medium.« less

Epidermal regulation of dermal fibroblast activity.

PubMed

Garner, W L

1998-07-01

Although the association between delayed burn wound healing and subsequent hypertrophic scar formation is well-established, the mechanism for this relationship is unknown. Unhealed burn wounds lack an epidermis, suggesting a possible regulatory role for the epidermis in controlling dermal fibroblast matrix synthesis. Therefore, we examined the effect of epidermal cells and media conditioned by epidermal cells on fibroblast collagen synthesis and replication. Purified fibroblast and keratinocyte cell strains were developed from discarded normal adult human skin. Conditioned media were created by incubation of cytokine-free and serum-free medium with either confluent fibroblast or keratinocyte cultures for 18 hours (n = 3). Nearly confluent fibroblast cultures were exposed for 48 hours to graded concentrations of either unconditioned medium (control), conditioned medium, or varying numbers of keratinocytes. Replication was quantified by the incorporation of 3H-thymidine. Collagen synthesis was measured by the incorporation of 3H-proline into collagenase-sensitive protein. Data were compared using analysis of variance (ANOVA) and linear regression. Keratinocyte conditioned medium induced a significant increase in replication (n = 3) (p = 0.004) and a decrease in collagen synthesis (n = 6) (p < 0.001). In contrast, neither fibroblast conditioned medium nor control medium had an effect on fibroblast replication or collagen synthesis. Co-culture of fibroblast with a graded number of keratinocytes similarly decreased collagen synthesis (n = 6) (p < 0.001). Dermal fibroblast collagen synthesis appears to be regulated by a soluble keratinocyte product. This result suggests a mechanism for the clinical observation that unhealed burn wounds, which lack the epidermis, demonstrate excess collagen production and scar. Clinical strategies to decrease hypertrophic scar should include an attempt at early wound closure with skin grafting or the application of cultured epithelial autografts.

Modelling of the Nutrient Medium for Plants Cultivation in Spaceflight

NASA Astrophysics Data System (ADS)

Nechitailo, Galina S.

2016-07-01

MODELLING OF THE NUTRIENT MEDIUM FOR PLANTS CULTIVATION IN SPACEFLIGHT Nechitajlo G.S.*, Rakhmetova A.A.**, Bogoslovskaja O.A.**, Ol'hovskay I.P.**, Glushchenko N.N.** *Emanuel Institute of Biochemical Physics of Russian Academy of Sciences (IBCP RAS) mail: spacemal@mail.ru **V.L. Talrose Institute for Energy Problems of Chemical Physics of Russian Academy of Science (INEPCP RAS) mail: nnglu@ mail.ru The valuable life and fruitful activity of cosmonauts and researchers in conditions of spaceflights and prolonged work at space stations are only possible with creating life area providing fresh air, natural food, comfortable psychological conditions, etc. The solution of that problem under space conditions seems impossible without use of high nano- and biotechnologies for plants growth. A priority should be given not only to choose species of growth plants in space, but also to improve conditions for their growth which includes optimal nourishing components for plants, preparation of nutrient mediums, illumination and temperature. We are deeply convinced that just manipulations with growing conditions for cultivated plants, but not genes changes, is a guarantee of success in the decision of this problem. For improving the method of plants growing on the artificial nutrient medium with balanced content of components, being necessary for growth and development of plants, we added essential metal elements: Fe, Zn, Cu - in an electroneutral state in the form of nanoparticles instead of sulfates or other easily dissolving salts. Nanoparticulated metals are known to have a number of advantages in comparison with salts: metals in an electroneutral form are characterized with the prolonged and multifunctional action, low toxicity per se and appearing to be much below the toxicity of the same metals in the ionic forms, accumulation as a reserve being used in biotic dozes, active distribution in bodies and organs of plants and stimulation of vital processes. A high reactivity of nanoparticles and their active interactions with components of a nutrient medium demands development of certain technological solutions for conservation of activity potential of nanoparticles in nutrient mediums. Thus, we have elaborated and created the artificial nutrient medium having balanced structure of components and assuring successful plants cultivation in conditions of spaceflight.

Classifying performance impairment in response to sleep loss using pattern recognition algorithms on single session testing

PubMed Central

St. Hilaire, Melissa A.; Sullivan, Jason P.; Anderson, Clare; Cohen, Daniel A.; Barger, Laura K.; Lockley, Steven W.; Klerman, Elizabeth B.

2012-01-01

There is currently no “gold standard” marker of cognitive performance impairment resulting from sleep loss. We utilized pattern recognition algorithms to determine which features of data collected under controlled laboratory conditions could most reliably identify cognitive performance impairment in response to sleep loss using data from only one testing session, such as would occur in the “real world” or field conditions. A training set for testing the pattern recognition algorithms was developed using objective Psychomotor Vigilance Task (PVT) and subjective Karolinska Sleepiness Scale (KSS) data collected from laboratory studies during which subjects were sleep deprived for 26 – 52 hours. The algorithm was then tested in data from both laboratory and field experiments. The pattern recognition algorithm was able to identify performance impairment with a single testing session in individuals studied under laboratory conditions using PVT, KSS, length of time awake and time of day information with sensitivity and specificity as high as 82%. When this algorithm was tested on data collected under real-world conditions from individuals whose data were not in the training set, accuracy of predictions for individuals categorized with low performance impairment were as high as 98%. Predictions for medium and severe performance impairment were less accurate. We conclude that pattern recognition algorithms may be a promising method for identifying performance impairment in individuals using only current information about the individual’s behavior. Single testing features (e.g., number of PVT lapses) with high correlation with performance impairment in the laboratory setting may not be the best indicators of performance impairment under real-world conditions. Pattern recognition algorithms should be further tested for their ability to be used in conjunction with other assessments of sleepiness in real-world conditions to quantify performance impairment in response to sleep loss. PMID:22959616

Accumulation and chemical states of radiocesium by fungus Saccharomyces cerevisiae

NASA Astrophysics Data System (ADS)

Ohnuki, Toshihiko; Sakamoto, Fuminori; Kozai, Naofumi; Yamasaki, Shinya; Yu, Qianqian

2014-05-01

After accident of Fukushima Daiichi Nuclear Power Plant, the fall-out radiocesium was deposited on the ground. Filamentous fungus is known to accumulate radiocesium in environment, even though many minerals are involved in soil. These facts suggest that fungus affect the migration behavior of radiocesium in the environment. However, accumulation mechanism of radiocesium by fungus is not understood. In the present study, accumulation and chemical states change of Cs by unicellular fungus of Saccharomyces cerevisiae have been studied to elucidate the role of microorganisms in the migration of radiocesium in the environment. Two different experimental conditions were employed; one is the accumulation experiments of radiocesium by S. cerevisiae from the agar medium containing 137Cs and a mineral of zeolite, vermiculite, smectite, mica, or illite. The other is the experiments using stable cesium to examine the chemical states change of Cs. In the former experiment, the cells were grown on membrane filter of 0.45 μm installed on the agar medium. After the grown cells were weighed, radioactivity in the cells was measured by an autoradiography technique. The mineral weight contents were changed from 0.1% to 1% of the medium. In the latter experiment, the cells were grown in the medium containing stable Cs between 1 mM and 10mM. The Cs accumulated cells were analyzed by SEM-EDS and EXAFS. The adsorption experiments of cesium by the cells under resting condition were also conducted to test the effect of cells metabolic activity. Without mineral in the medium, cells of S. cerevisiae accumulated 1.5x103 Bq/g from the medium containing 137Cs of 2.6x102 Bq/g. When mineral was added in the medium, concentration of 137Cs in the cells decreased. The concentration of 137Cs in the cells from the medium containing different minerals were in the following order; smectite, illite, mica > vermiculite > zeolite. This order was nearly the same as the inverse of distribution coefficient of mineral for 137Cs in the medium solution. The concentration of 137Cs in the cells lowered in the medium containing higher mineral content. These results indicate that radiocesium was competively accumulated in the cells with minerals in the soil. Higher concentration of stable Cs was accumulated in the cells in the metabolically active condition than in the resting cells condition. XAFS analyses showed that the k3-weighted extended-XAFS functions and the radial structural function of Cs accumulated by the cells in the metabolically active condition were similar to those in the resting condition, indicating that chemical states of the accumulated Cs were nearly the same between both conditions. These results indicate that the fungus accumulates radiocesium by competitively with minerals in the soils, and performs higher retardation of the migration of Cs in the metabolically active condition than the resting one. A part of this study is the results of "Multidisciplinary investigation on radiocesium fate and transport for safety assessment for interim storage and disposal of heterogeneous waste" carried out under the Initiatives for Atomic Energy Basic and Generic Strategic Research by the Ministry of Education, Culture, Sports, Science and Technology of Japan.

Heparin affin regulatory peptide/pleiotrophin negatively affects diverse biological activities in C6 glioma cells.

PubMed

Parthymou, Anastasia; Lampropoulou, Evgenia; Mikelis, Constantinos; Drosou, Georgia; Papadimitriou, Evangelia

2008-01-01

Heparin affin regulatory peptide (HARP) or pleiotrophin seems to be involved in the progression of several tumors of diverse origin. In this study, we tried to determine the role of HARP in rat C6 glioma cells by using an antisense strategy for inhibition of HARP expression. Decrease of the expression of endogenous HARP in C6 cells (AS-C6 cells) significantly increased proliferation, migration, and anchorage-independent growth of cells. Implantation of AS-C6 cells onto chicken embryo chorioallantoic membranes resulted in a significant increase of tumor-induced angiogenesis compared with that induced by non-transfected or C6 cells transfected with the plasmid alone (PC-C6 cells). In the same line, conditioned medium from AS-C6 cells significantly increased endothelial cell proliferation, migration, and tube formation in vitro compared with the effect of conditioned medium from C6 or PC-C6 cells. Interestingly, vascular endothelial growth factor (VEGF) induced C6 cell proliferation and migration, and SU1496, a selective inhibitor of VEGF receptor 2 (VEGFR2), blocked increased glioma cell growth, migration, and angiogenicity observed in AS-C6 cell cultures. The above results seem to be due to a direct interaction between HARP and VEGF in the culture medium of C6 and PC-C6 cells, while AS-C6 cells secreted comparable amounts of VEGF that do not interact with HARP. Collectively, these data suggest that HARP negatively affects diverse biological activities in C6 glioma cells, mainly due to binding of HARP to VEGF, which may sequester secreted VEGF from signalling through VEGFR2.

In vitro conservation and sustained production of breadfruit ( Artocarpus altilis, Moraceae): modern technologies for a traditional tropical crop

NASA Astrophysics Data System (ADS)

Murch, Susan J.; Ragone, Diane; Shi, Wendy Lei; Alan, Ali R.; Saxena, Praveen K.

2008-02-01

Breadfruit ( Artocarpus altilis, Moraceae) is a traditionally cultivated, high-energy, high-yield crop, but widespread use of the plant for food is limited by poor quality and poor storage properties of the fruit. A unique field genebank of breadfruit species and cultivars exists at the National Tropical Botanical Garden in the Hawaiian Islands and is an important global resource for conservation and sustainable use of breadfruit. However, this plant collection could be damaged by a random natural disaster such as a hurricane. We have developed a highly efficient in vitro plant propagation system to maintain, conserve, mass propagate, and distribute elite varieties of this important tree species. Mature axillary shoot buds were collected from three different cultivars of breadfruit and proliferated using a cytokinin-supplemented medium. The multiple shoots were maintained as stock cultures and repeatedly used to develop whole plants after root differentiation on a basal or an auxin-containing medium. The plantlets were successfully grown under greenhouse conditions and were reused to initiate additional shoot cultures for sustained production of plants. Flow cytometry was used to determine the nuclear deoxyribonucleic acid content and the ploidy status of the in vitro grown population. The efficacy of the micropropagation protocols developed in this study represents a significant advancement in the conservation and sustained mass propagation of breadfruit germplasm in a controlled environment free from contamination.

The Secret Life of Exosomes: What Bees Can Teach Us About Next-Generation Therapeutics.

PubMed

Marbán, Eduardo

2018-01-16

Mechanistic exploration has pinpointed nanosized extracellular vesicles, known as exosomes, as key mediators of the benefits of cell therapy. Exosomes appear to recapitulate the benefits of cells and more. As durable azoic entities, exosomes have numerous practical and conceptual advantages over cells. Will cells end up just being used to manufacture exosomes, or will they find lasting value as primary therapeutic agents? Here, a venerable natural process-the generation of honey-serves as an instructive parable. Flowers make nectar, which bees collect and process into honey. Cells make conditioned medium, which laboratory workers collect and process into exosomes. Unlike flowers, honey is durable, compact, and nutritious, but these facts do not negate the value of flowers themselves. The parallels suggest new ways of thinking about next-generation therapeutics. Copyright © 2018 American College of Cardiology Foundation. Published by Elsevier Inc. All rights reserved.

ITO Modification for Efficient Inverted Organic Solar Cells.

PubMed

Susarova, Diana K; Akkuratov, Alexander V; Kukharenko, Andrey I; Cholakh, Seif O; Kurmaev, Ernst Z; Troshin, Pavel A

2017-10-03

We demonstrate a facile approach to designing transparent electron-collecting electrodes by depositing thin layers of medium and low work function metals on top of transparent conductive metal oxides (TCOs) such as ITO and FTO. The modified electrodes were fairly stable for months under ambient conditions and maintained their electrical characteristics. XPS spectroscopy data strongly suggested integration of the deposited metal in the TCO structure resulting in additional doping of the conducting oxide at the interface. Kelvin probe microscopy measurements revealed a significant decrease in the ITO work function after modification. Organic solar cells based on three different conjugated polymers have demonstrated state of the art performances in inverted device geometry using Mg- or Yb-modified ITO as electron collecting electrode. The simplicity of the proposed approach and the excellent ambient stability of the modified ITO electrodes allows one to expect their wide utilization in research laboratories and electronic industry.

Adult hippocampus derived soluble factors induce a neuronal-like phenotype in mesenchymal stem cells.

PubMed

Rivera, Francisco J; Sierralta, Walter D; Minguell, Jose J; Aigner, Ludwig

2006-10-02

Bone marrow-derived mesenchymal stem cells (MSCs) are not restricted in their differentiation fate to cells of the mesenchymal lineage. They acquire a neural phenotype in vitro and in vivo after transplantation in the central nervous system. Here we investigated whether soluble factors derived from different brain regions are sufficient to induce a neuronal phenotype in MSCs. We incubated bone marrow-derived MSCs in conditioned medium (CM) derived from adult hippocampus (HCM), cortex (CoCM) or cerebellum (CeCM) and analyzed the cellular morphology and the expression of neuronal and glial markers. In contrast to muscle derived conditioned medium, which served as control, conditioned medium derived from the different brain regions induced a neuronal morphology and the expression of the neuronal markers GAP-43 and neurofilaments in MSCs. Hippocampus derived conditioned medium had the strongest activity. It was independent of NGF or BDNF; and it was restricted to the neuronal differentiation fate, since no induction of the astroglial marker GFAP was observed. The work indicates that soluble factors present in the brain are sufficient to induce a neuronal phenotype in MSCs.

76 FR 18264 - Proposed Collection; Comment Request

Federal Register 2010, 2011, 2012, 2013, 2014

2011-04-01

..., Copies Available From: Securities and Exchange Commission, Office of Investor Education and Advocacy... resumption of quotations in a quotation medium by a broker-dealer for over-the- counter (``OTC'') securities... quotation for publication, in a quotation medium unless they have reviewed specified information concerning...

Electric filter with movable belt electrode

DOEpatents

Bergman, W.

1983-09-20

A method and apparatus for removing airborne contaminants entrained in a gas or airstream includes an electric filter characterized by a movable endless belt electrode, a grounded electrode, and a filter medium sandwiched there between. Inclusion of the movable, endless belt electrode provides the driving force for advancing the filter medium through the filter, and reduces frictional drag on the filter medium, thereby permitting a wide choice of filter medium materials. Additionally, the belt electrode includes a plurality of pleats in order to provide maximum surface area on which to collect airborne contaminants. 4 figs.

Electric filter with movable belt electrode

DOEpatents

Bergman, Werner

1983-01-01

A method and apparatus for removing airborne contaminants entrained in a gas or airstream includes an electric filter characterized by a movable endless belt electrode, a grounded electrode, and a filter medium sandwiched therebetween. Inclusion of the movable, endless belt electrode provides the driving force for advancing the filter medium through the filter, and reduces frictional drag on the filter medium, thereby permitting a wide choice of filter medium materials. Additionally, the belt electrode includes a plurality of pleats in order to provide maximum surface area on which to collect airborne contaminants.

Porous media heat transfer for injection molding

DOEpatents

Beer, Neil Reginald

2016-05-31

The cooling of injection molded plastic is targeted. Coolant flows into a porous medium disposed within an injection molding component via a porous medium inlet. The porous medium is thermally coupled to a mold cavity configured to receive injected liquid plastic. The porous medium beneficially allows for an increased rate of heat transfer from the injected liquid plastic to the coolant and provides additional structural support over a hollow cooling well. When the temperature of the injected liquid plastic falls below a solidifying temperature threshold, the molded component is ejected and collected.

77 FR 35679 - Agency Information Collection Activities: Final Collection; Comment Request

Federal Register 2010, 2011, 2012, 2013, 2014

2012-06-14

... EXPORT-IMPORT BANK OF THE UNITED STATES [Public Notice 2012-0136] Agency Information Collection Activities: Final Collection; Comment Request AGENCY: Export-Import Bank of the U.S. ACTION: Submission for... Financial Institution of Medium Term Export Credit Insurance Policy. SUMMARY: The Export-Import Bank of the...

Conditioned medium: a new alternative for cryopreservation of equine umbilical cord mesenchymal stem cells.

PubMed

Maia, Leandro; Dias, Marianne Camargos; de Moraes, Carolina Nogueira; de Paula Freitas-Dell'Aqua, Camila; da Mota, Ligia S L Silveira; Santiloni, Valquíria; da Cruz Landim-Alvarenga, Fernanda

2017-03-01

Cryopreservation is a feasible alternative to maintaining several cell lines, particularly for immediate therapeutic use, transportation of samples, and implementation of new in vitro studies. This work parts from the hypothesis that the medium of cryopreservation composed by 90% of conditioned medium (CM) supports cryopreservation of equine umbilical cord intervascular matrix mesenchymal stem cells (UCIM-MSCs), allowing the maintenance of the biological properties for the establishment of cell banks intended for therapeutic use and in vitro studies. Thus, we evaluated the viability, apoptosis/necrosis rates, immunophenotypic profile (IP), chromosomal stability, clonicity, and differentiation potential of UCIM-MSCs cryopreserved with four different mediums (with FBS: M1, M3, M4 and without FBS: M2). After 3 months of cryopreservation, samples were thawed and analyzed. The potential of differentiation in the mesodermal lineages, clonicity, and the chromosomal stability were maintained after cryopreservation of UCIM-MSCs with medium containing FBS. Changes (P < 0.05) at IP for some markers were observed at cells cryopreserved with medium M1-M3. Only the UCIM-MSCs cryopreserved with the CM (M4) had similar viability post-thaw (P = 0.23) when compared with fresh cells. We proved the hypothesis that the medium of cryopreservation containing CM supports the cryopreservation of UCIM-MSCs, at the experimental conditions, being the medium that better maintains the biological characteristics observed at fresh cells. Thus, future studies of UCIM-MSCs secretome should be conducted to better understand the beneficial and protective effects of the CM during the freezing process. © 2017 International Federation for Cell Biology.

Pericellular oxygen concentration of cultured primary human trophoblasts

PubMed Central

Chen, Baosheng; Longtine, Mark S.; Nelson, D. Michael

2012-01-01

Introduction Oxygen is pivotal in placental development and function. In vitro culture of human trophoblasts provides a useful model to study this phenomenon, but a hotly debated issue is whether or not the oxygen tension of the culture conditions mimics in vivo conditions. We tested the hypothesis that ambient oxygen tensions in culture reflect the pericellular oxygen levels. Methods We used a microelectrode oxygen sensor to measure the concentration of dissolved oxygen in the culture medium equilibrated with 21%, 8% or <0.5% oxygen. Results The concentration of oxygen in medium without cells resembled that in the ambient atmosphere. The oxygen concentration present in medium bathing trophoblasts was remarkably dependent on the depth within the medium where sampling occurred, and the oxygen concentration within the overlying atmosphere was not reflected in medium immediately adjacent to the cells. Indeed, the pericellular oxygen concentration was in a range that most would consider severe hypoxia, at ≤ 0.6% oxygen or about 4.6 mm Hg, when the overlying atmosphere was 21% oxygen. Conclusions We conclude that culture conditions of 21% oxygen are unable to replicate the pO2 of 40–60 mm Hg commonly attributed to the maternal blood in the intervillous space in the second and third trimesters of pregnancy. We further surmise that oxygen atmospheres in culture conditions between 0.5% and 21% provide different oxygen fluxes in the immediate pericellular environment yet can still yield insights into the responses of human trophoblast to different oxygen conditions. PMID:23211472

Basic FGF Support of Human Embryonic Stem Cell Self-Renewal

PubMed Central

Levenstein, Mark E.; Ludwig, Tenneille E.; Xu, Ren-He; Llanas, Rachel A.; VanDenHeuvel-Kramer, Kaitlyn; Manning, Daisy; Thomson, James A.

2015-01-01

Human embryonic stem (ES) cells have most commonly been cultured in the presence of basic FGF (FGF2) either on fibroblast feeder layers or in fibroblast-conditioned medium. Recently, it has been reported that elevated concentrations of FGF2 permit the culture of human ES cells in the absence of fibroblasts or fibroblast-conditioned medium. Here we compare the ability of unconditioned medium (UM) supplemented with 4, 24, 40, 80, 100 and 250 ng/ml FGF2 to sustain low-density human ES cell cultures through multiple passages. In these stringent culture conditions, 4, 24, and 40 ng/ml FGF2 failed to sustain human ES cells through three passages, but 100 ng/ml sustained human ES cells with an effectiveness comparable to conditioned medium (CM). Two human ES cell lines (H1 and H9) were maintained for up to 164 population doublings (7 and 4 months) in UM supplemented with 100 ng/ml FGF2. After prolonged culture the cells formed teratomas when injected into SCID-beige mice, and expressed markers characteristic of undifferentiated human ES cells. We also demonstrate that FGF2 is degraded more rapidly in UM than in CM, partly explaining the need for higher concentrations of FGF2 in UM. These results further facilitate the large-scale, routine culture of human ES cells, and suggest that fibroblasts and fibroblast-conditioned medium sustain human ES cells in part by stabilizing FGF signaling above a critical threshold. PMID:16282444

The Approval-Built Collection in the Medium-Sized Academic Library

ERIC Educational Resources Information Center

DeVilbiss, Mary Lee

1975-01-01

Compares the approval-built collection with the collection which is created when traditional select and order procedures are used. The study was limited to 1974 imprints in four subject areas and utilized the services of two major vendors. (Author)

Lidocaine and Pain Management in the Emergency Department: A Review Article

PubMed Central

Golzari, Samad EJ; Soleimanpour, Hassan; Mahmoodpoor, Ata; Safari, Saeid; Ala, Alireza

2014-01-01

Context: In the present review, the analgesic effects of lidocaine in acute or chronic painful conditions in the emergency department are discussed. Lidocaine, as a medium-acting local anesthetic with short onset time, is well-recognized, not only as a valuable medication for numerous neuropathic pain conditions, but also for the management of both acute and chronic pain. Evidence Acquisition: Research studies related to the different applications of lidocaine in the emergency department were collected from different databases including Cochrane library, Medline (Ovid) and PubMed. The pooled data were categorized, summarized and finally compared. Results: Our study revealed that lidocaine is broadly used in various therapeutic approaches for different types of pain, such as visceral/central pain, renal colic etc., in the emergency department. Conclusions: The antinociceptive properties of lidocaine are derived from multifaceted mechanisms, turning it into a medication that is safe to administer via different routes which makes it available for use in a variety of medical conditions. PMID:24660158

[Killing effect of polymorphonuclear neutrophils on Trichomonas vaginalis].

PubMed

Zhao, Jian-Ling; Gao, Xing-Zheng; Qu, Ming

2008-10-30

To study the killing effect of polymorphonuclear neutrophils (PMNs) on Trichomonas vaginalis. The vaginal secretion from a patient with vaginitis was incubated in the liver infusion liquid medium to get T. vaginalis. One ml serum was collected from the patient and heated for 30 min at 56 degrees C to inactivate complement in serum, and was absorbed three times with the parasites at 0 degree C to make the serum free of antibodies. PMNs were separated from the patient's blood and purified with density gradient centrifugation and polymer accelerating sedimentation. NBT and safranin O were used to stain the sample. The interaction between PMNs and the parasites was observed under microscope. 300 trichomonads and 3x10(4) PMNs were incubated for 10, 20, 30, 40, 50, 60 minutes under the conditions of aerobic or anaerobic, with superoxide dismutase (SOD) and catalase (CAT) or without SOD and CAT, and with complement or without complement. They were then inoculated in solid medium for another five days under the anaerobic condition, and surviving organisms were enumerated. PMNs were observed to surround and kill a single trichomonad. In the petri-dish containing PMNs, the surviving rate of the parasites in anaerobic condition was 85%, only 3% in aerobic condition (P<0.01). SOD and CAT reduced the killing effect of PMNs, with a surviving rate of 98% and 94% respectively after 60 min incubation. Without SOD and CAT, the surviving rate is only 2% (P<0.05). PMNs in the serum without antibodies killed all the parasites, while the complement-inactivated serum fail to kill them. The trichomonacidal activity of PMNs relies on the presence of oxygen and complement in the serum of patient.

A simple and cost effective liquid culture system for the micropropagation of two commercially important apple rootstocks.

PubMed

Mehta, Mohina; Ram, Raja; Bhattacharya, Amita

2014-07-01

The two commercially important apple rootstocks i.e., MM106 and B9 were micropropagated using a liquid culture system. Three different strengths of 0.8% agar solidified PGR free basal MS medium were first tested to optimize the culture media for both the rootstocks. Full strength medium (MS0) supported maximum in vitro growth, multiplication, rooting and survival under field conditions as opposed to quarter and half strength media. When three different volumes of liquid MS0 were tested, highest in vitro growth, multiplication, rooting and also survival under field conditions were achieved in 20 mL liquid MS0. The cost of one litre of liquid medium was also reduced by 8 times to Rs. 6.29 as compared to solid medium. The cost of 20 mL medium was further reduced to Rs. 0.125.

Gamma-ray line afterglow from burst environments

NASA Technical Reports Server (NTRS)

Fencl, H. S.; Boyd, R. N.; Hartmann, Dieter

1992-01-01

We consider photoerosion and direct pair production in a medium surrounding a gamma-ray burst. The resulting secondary gamma-rays may provide diagnostic tools of these environments and, in turn, of the nature of the bursters themselves. In some instances short-lived nuclides are formed; the beta-delayed gamma-rays produced from their decays provide the signatures of the photoerosion. In addition, annihilation radiation produced from positrons resulting from direct pair production is related to the plasma conditions in the medium. We investigate the plausibility of detecting the various radiations. Under extremely favorable conditions, the photoerosion afterglow might be detectable with the present generation of detectors. However, the positron annihilation line should be detectable under a fairly wide range in the conditions of the medium.

Stimulation of plasmin activity in cultured human fibroblast cells by Porphyromonas endodontalis.

PubMed

Oikawa, T; Ogura, N; Akiba, M; Abiko, Y; Takiguchi, H; Izumi, H

1993-09-01

1. Plasmin activity in the conditioned medium of Gin-1 cells, a human gingival fibroblast cell line, was stimulated by Porphyromonas endodontalis, a putative pathogen of oral submucous abscesses, in a time- and dose-dependent manner. 2. P. endodontalis stimulated the activity of plasminogen activator in both the conditioned medium and the cell lysate. The plasminogen activator in Gin-1 cells was approx. 50 kDa by zymography. 3. The conditioned medium of Gin-1 cells exposed to P. endodontalis stimulated the conversion of human serum prekallikrein to kallikrein. 4. These results suggested that P. endodontalis stimulates the plasminogen activator-plasmin system in Gin-1 cells, and that activated plasmin plays a role in the progress of periodontal tissue inflammation.

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.

Evaluation of sampling and analytical methods for nicotine and polynuclear aromatic hydrocarbon in indoor air. Final report, 1 February 1987-30 March 1987

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

Chuang, J.C.; Kuhlman, M.R.; Hannan, S.W.

1987-11-01

The objective of this project was to evaluate a potential collection medium, XAD-4 resin, for collecting nicotine and polynuclear aromatic hydrocarbon (PAH) and to determine whether one collection system and one analytical method will allow quantification of both compound classes in air. The extraction efficiency study was to determine the extraction method to quantitatively remove nicotine and PAH from XAD-4 resin. The results showed that a two-step Soxhlet extraction consisting of dichloromethane followed by ethyl acetate resulted in the best recoveries for both nicotine and PAH. In the sampling efficiency study, XAD-2 and XAD-4 resin were compared, in parallel, formore » collection of PAH and nicotine. Quartz fiber filters were placed upstream of both adsorbents to collect particles. Prior to sampling, both XAD-2 and XAD-4 traps were spiked with known amounts (2 microgram) of perdeuterated PAH and D3-nicotine. The experiments were performed with cigarette smoking and nonsmoking conditions. The spiked PAH were retained well in both adsorbents after exposure to more than 300 cu. m. of indoor air. The spiked XAD-4 resin gave higher recoveries for D3-nicotine than did the spiked XAD-2 resin. The collection efficiency for PAH for both adsorbents is very similar but higher levels of nicotine were collected on XAD-4 resin.« less

Radioactive waste management and practice in Bangladesh

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

Mollah, A.S.; Rahman, M.M.

1993-12-31

A large amount of low- and medium-level radioactive wastes are being generated in different parts of Bangladesh. The solid wastes are being collected in steel containers and liquid wastes are collected in plastic carboys and drums. Gaseous Ar-41 is discharged into the atmosphere through the 25 m height stack under controlled conditions after proper monitoring. The solid radioactive wastes collected are approximately 5 m{sup 3} (1988--1992) with gross beta-gamma surface dose rates from 0.30 {micro}Sv/h to 250 {micro}Sv/h. The liquid radioactive wastes are approximately 200 liters (1988--1992) with gross-beta-gamma surface dose rates from 0.30 {micro}Sv/h to 1 mSv/h. The solidmore » and liquid wastes presently being collected are mostly short lived and low level and safely stored according to international safety codes of practice. Radioactive waste packages collected during the 5-yrs study totaled 16, representing a collective volume of {approximately} 7.5 m{sup 3}. The problem of management of radioactive waste in Bangladesh is not so serious at present because the wastes arising are small now. A computerized data base has been developed to document inventory of all radioactive waste arising in the country. The current practices of collection, handling, safe storage and management of the radioactive wastes are reported in this paper.« less

Mesh Intercomparisons of Fog Water Collected Yield Insight Into the Nature of Fog-Drip Collection Mechanisms

NASA Astrophysics Data System (ADS)

Fernandez, D.; Torregrosa, A.; Weiss-Penzias, P. S.; Oliphant, A. J.; Dodge, C.; Bowman, M.; Wilson, S.; Mairs, A. A.; Gravelle, M.; Barkley, T.

2016-12-01

At multiple sites across central CA, several passive fog water collectors have been deployed for the past 3 years. All of the sites employ standard Raschel polypropylene mesh as the fog collection medium and five of them also integrated a novel polypropylene mesh of German manufacture with a 3-dimensional internal structure. Additionally, six metal mesh manufactured by McMaster-Carr of various hole sizing were coated with a POSS-PEMA substance at the Massachusetts Institute of Technology and deployed in parallel with the Raschel mesh at six distinct locations. Finally, fluorine-free versions of the POSS-PEMA substance were generated by NBD Nanotechnology and coated on a much finer mesh substrate. Three of those and one control (uncoated mesh) were deployed at one of the fog collection sites for one season, along with a standard Raschel mesh. Preliminary results from one intercomparison from just one pair of mesh over two seasons seem to reveal a wind speed and also, possibly, a droplet-size dependence on the fog collection efficiency for the mesh. This study will continue to intercompare the various mesh in conjunction with the wind speed and direction data. If a collection efficiency dependence on mesh size or coating is confirmed, it may point to interesting and relevant mechanisms for fog droplet capture and collection hitherto unobserved in field conditions.

Medium components and culture conditions affect the thermotolerance of aerial conidia of fungal biocontrol agent Beauveria bassiana.

PubMed

Ying, S-H; Feng, M-G

2006-09-01

To produce more thermotolerable conidia of Beauveria bassiana, a well-known fungal biocontrol agent, by optimizing the medium components and culture conditions. The conidia produced on media including 0.5-6% glucose, sucrose or starch as carbon source and 50-300-microg ml(-1) Cu2+, Zn2+, Mn2+ or Fe3+ as additive to Sabouraud dextrose medium at 15-30 degrees C, pH 4-8 or KCl-adjusted water availabilities were exposed to 30-min wet heat stress at 48 degrees C. The medium components for conidial production with greatly enhanced thermotolerance included 4% glucose as optimum or 1% starch as alternative for the carbon source and < or =50-microg ml(-1) Mn2+ for the metal additive. The culture conditions were optimized as 25 degrees C and pH 5-6. Conidial thermotolerance decreased remarkably when sucrose and Fe3+ or Cu2+ were used in the cultures, but altered slightly when 50-200-microg ml(-1) Zn2+ were included. The tolerance of B. bassiana conidia to the thermal stress was significantly affected by the medium composition and culture conditions under which the conidia were produced. Proper treatment of small grains as mass production substrates for more glucose release and supplement of glucose or 50-microg ml(-1) Mn2+ are possible means to enhancing conidial thermotolerance and field persistence for improved insect control.

Practical methods for handling human periodontal ligament stem cells in serum-free and serum-containing culture conditions under hypoxia: implications for regenerative medicine.

PubMed

Murabayashi, Dai; Mochizuki, Mai; Tamaki, Yuichi; Nakahara, Taka

2017-07-01

Stem cell-based therapies depend on the reliable expansion of patient-derived mesenchymal stem cells (MSCs) in vitro. The supplementation of cell culture media with serum is associated with several risks; accordingly, serum-free media are commercially available for cell culture. Furthermore, hypoxia is known to accelerate the expansion of MSCs. The present study aimed to characterize the properties of periodontal ligament-derived MSCs (PDLSCs) cultivated in serum-free and serum-containing media, under hypoxic and normoxic conditions. Cell growth, gene and protein expression, cytodifferentiation potential, genomic stability, cytotoxic response, and in vivo hard tissue generation of PDLSCs were examined. Our findings indicated that cultivation in serum-free medium does not affect the MSC phenotype or chromosomal stability of PDLSCs. PDLSCs expanded in serum-free medium exhibited more active growth than in fetal bovine serum-containing medium. We found that hypoxia does not alter the cell growth of PDLSCs under serum-free conditions, but inhibits their osteogenic and adipogenic cytodifferentiation while enabling maintenance of their multidifferentiation potential regardless of the presence of serum. PDLSCs expanded in serum-free medium were found to retain common MSC characteristics, including the capacity for hard tissue formation in vivo. However, PDLSCs cultured in serum-free culture conditions were more susceptible to damage following exposure to extrinsic cytotoxic stimuli than those cultured in medium supplemented with serum, suggesting that serum-free culture conditions do not exert protective effects against cytotoxicity on PDLSC cultures. The present work provides a comparative evaluation of cell culture in serum-free and serum-containing media, under hypoxic and normoxic conditions, for applications in regenerative medicine.

A Rapid Filter Insert-based 3D Culture System for Primary Prostate Cell Differentiation

PubMed Central

Tricoli, Lucas; Berry, Deborah L.; Albanese, Chris

2018-01-01

Conditionally reprogrammed cells (CRCs) provide a sustainable method for primary cell culture and the ability to develop extensive “living biobanks” of patient derived cell lines. For many types of epithelial cells, various three dimensional (3D) culture approaches have been described that support an improved differentiated state. While CRCs retain their lineage commitment to the tissue from which they are isolated, they fail to express many of the differentiation markers associated with the tissue of origin when grown under normal two dimensional (2D) culture conditions. To enhance the application of patient-derived CRCs for prostate cancer research, a 3D culture format has been defined that enables a rapid (2 weeks total) luminal cell differentiation in both normal and tumor-derived prostate epithelial cells. Herein, a filter insert-based format is described for the culturing and differentiation of both normal and malignant prostate CRCs. A detailed description of the procedures required for cell collection and processing for immunohistochemical and immunofluorescent staining are provided. Collectively the 3D culture format described, combined with the primary CRC lines, provides an important medium- to high- throughput model system for biospecimen-based prostate research. PMID:28287583

Analysis of the nutritional status of algae by Fourier transform infrared chemical imaging

NASA Astrophysics Data System (ADS)

Hirschmugl, Carol J.; Bayarri, Zuheir-El; Bunta, Maria; Holt, Justin B.; Giordano, Mario

2006-09-01

A new non-destructive method to study the nutritional status of algal cells and their environments is demonstrated. This approach allows rapid examination of whole cells without any or little pre-treatment providing a large amount of information on the biochemical composition of cells and growth medium. The method is based on the analysis of a collection of infrared (IR) spectra for individual cells; each spectrum describes the biochemical composition of a portion of a cell; a complete set of spectra is used to reconstruct an image of the entire cell. To obtain spatially resolved information synchrotron radiation was used as a bright IR source. We tested this method on the green flagellate Euglena gracilis; a comparison was conducted between cells grown in nutrient replete conditions (Type 1) and on cells allowed to deplete their medium (Type 2). Complete sets of spectra for individual cells of both types were analyzed with agglomerative hierarchical clustering, leading to distinct clusters representative of the two types of cells. The average spectra for the clusters confirmed the similarities between the clusters and the types of cells. The clustering analysis, therefore, allows the distinction of cells of the same species, but with different nutritional histories. In order to facilitate the application of the method and reduce manipulation (washing), we analyzed the cells in the presence of residual medium. The results obtained showed that even with residual medium the outcome of the clustering analysis is reliable. Our results demonstrate the applicability FTIR microspectroscopy for ecological and ecophysiological studies.

Restoration of longitudinal laser tomography target image from inhomogeneous medium degradation under common conditions.

PubMed

Yi, WenJun; Wang, Ping; Fu, MeiCheng; Tan, JiChun; Zhu, Jubo; Li, XiuJian

2017-07-10

In order to overcome the shortages of the target image restoration method for longitudinal laser tomography using self-calibration, a more general restoration method through backscattering medium images associated with prior parameters is developed for common conditions. The system parameters are extracted from pre-calibration, and the LIDAR ratio is estimated according to the medium types. Assisted by these prior parameters, the degradation caused by inhomogeneous turbid media can be established with the backscattering medium images, which can further be used for removal of the interferences of turbid media. The results of simulations and experiments demonstrate that the proposed image restoration method can effectively eliminate the inhomogeneous interferences of turbid media and achieve exactly the reflectivity distribution of targets behind inhomogeneous turbid media. Furthermore, the restoration method can work beyond the limitation of the previous method that only works well under the conditions of localized turbid attenuations and some types of targets with fairly uniform reflectivity distributions.

Interaction effect of response medium and working memory capacity on creative idea generation

PubMed Central

Hao, Ning; Yuan, Huan; Cheng, Rui; Wang, Qing; Runco, Mark A.

2015-01-01

This study aimed to examine the interaction effect of response medium (i.e., write down ideas and orally report ideas) and working memory capacity (WMC) on creative idea generation. Participants (N = 90) with higher or lower WMC were asked to solve Alternative Uses Task (AUT) problems in the condition of writing down or speaking out ideas. The results showed that fluency of AUT performance was higher in the writing than in the speaking condition. Additionally, participants with higher WMC performed better on AUT fluency than those with lower WMC in the writing condition, while they showed no difference in the speaking condition. Moreover, level of cognitive demand fully mediated the effect of response medium on AUT fluency. Theoretically, these findings indicated the importance of WMC in creative idea generation, which supported the controlled-attention theory of creativity. Practical implications and future directions were discussed. PMID:26528227

Interaction effect of response medium and working memory capacity on creative idea generation.

PubMed

Hao, Ning; Yuan, Huan; Cheng, Rui; Wang, Qing; Runco, Mark A

2015-01-01

This study aimed to examine the interaction effect of response medium (i.e., write down ideas and orally report ideas) and working memory capacity (WMC) on creative idea generation. Participants (N = 90) with higher or lower WMC were asked to solve Alternative Uses Task (AUT) problems in the condition of writing down or speaking out ideas. The results showed that fluency of AUT performance was higher in the writing than in the speaking condition. Additionally, participants with higher WMC performed better on AUT fluency than those with lower WMC in the writing condition, while they showed no difference in the speaking condition. Moreover, level of cognitive demand fully mediated the effect of response medium on AUT fluency. Theoretically, these findings indicated the importance of WMC in creative idea generation, which supported the controlled-attention theory of creativity. Practical implications and future directions were discussed.

Effect of cultivating conditions on alpha-galactosidase production by a novel Aspergillus foetidus ZU-G1 strain in solid-state fermentation.

PubMed

Liu, Cai-qin; Chen, Qi-he; Cheng, Qian-jun; Wang, Jin-ling; He, Guo-qing

2007-05-01

The work is intended to achieve optimum culture conditions of alpha-galactosidase production by a mutant strain Aspergillus foetidus ZU-G1 in solid-state fermentation (SSF). Certain fermentation parameters involving moisture content, incubation temperature, cultivation period of seed, inoculum volume, initial pH value, layers of pledget, load size of medium and period of cultivation were investigated separately. The optimal cultivating conditions of alpha-galactosidase production in SSF were 60% initial moisture of medium, 28 degrees C incubation temperature, 18 h cultivation period of seed, 10% inoculum volume, 5.0 approximately 6.0 initial pH of medium, 6 layers of pledget and 10 g dry matter loadage. Under the optimized cultivation conditions, the maximum alpha-galactosidase production was 2 037.51 U/g dry matter near the 144th hour of fermentation.

Effect of cultivating conditions on α-galactosidase production by a novel Aspergillus foetidus ZU-G1 strain in solid-state fermentation

PubMed Central

Liu, Cai-qin; Chen, Qi-he; Cheng, Qian-jun; Wang, Jin-ling; He, Guo-qing

2007-01-01

The work is intended to achieve optimum culture conditions of α-galactosidase production by a mutant strain Aspergillus foetidus ZU-G1 in solid-state fermentation (SSF). Certain fermentation parameters involving moisture content, incubation temperature, cultivation period of seed, inoculum volume, initial pH value, layers of pledget, load size of medium and period of cultivation were investigated separately. The optimal cultivating conditions of α-galactosidase production in SSF were 60% initial moisture of medium, 28 °C incubation temperature, 18 h cultivation period of seed, 10% inoculum volume, 5.0~6.0 initial pH of medium, 6 layers of pledget and 10 g dry matter loadage. Under the optimized cultivation conditions, the maximum α-galactosidase production was 2 037.51 U/g dry matter near the 144th hour of fermentation. PMID:17542067

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.

Evaluation of genetic diversity in jackfruit (Artocarpus heterophyllus Lam.) based on amplified fragment length polymorphism markers.

PubMed

Shyamalamma, S; Chandra, S B C; Hegde, M; Naryanswamy, P

2008-07-22

Artocarpus heterophyllus Lam., commonly called jackfruit, is a medium-sized evergreen tree that bears high yields of the largest known edible fruit. Yet, it has been little explored commercially due to wide variation in fruit quality. The genetic diversity and genetic relatedness of 50 jackfruit accessions were studied using amplified fragment length polymorphism markers. Of 16 primer pairs evaluated, eight were selected for screening of genotypes based on the number and quality of polymorphic fragments produced. These primer combinations produced 5976 bands, 1267 (22%) of which were polymorphic. Among the jackfruit accessions, the similarity coefficient ranged from 0.137 to 0.978; the accessions also shared a large number of monomorphic fragments (78%). Cluster analysis and principal component analysis grouped all jackfruit genotypes into three major clusters. Cluster I included the genotypes grown in a jackfruit region of Karnataka, called Tamaka, with very dry conditions; cluster II contained the genotypes collected from locations having medium to heavy rainfall in Karnataka; cluster III grouped the genotypes in distant locations with different environmental conditions. Strong coincidence of these amplified fragment length polymorphism-based groupings with geographical localities as well as morphological characters was observed. We found moderate genetic diversity in these jackfruit accessions. This information should be useful for tree breeding programs, as part of our effort to popularize jackfruit as a commercial crop.

[Experimental study on co-culture of salivary adenoid cystic carcinoma cells and ganglia].

PubMed

Gu, Ling; Bu, Rong-fa; Wang, Dong-sheng; E, Ling-ling; Zhu, Guo-xiong

2012-01-01

To construct the co-culture models of salivarya denoid cystic carcinoma (SACC) cells and dorsal root ganglia (DRG) of chickens and investigate the promotive effects of SACC on neural tissue. Glass-base culture dish was adopted to construct co-culture model of SACC-83 cells and DRG. SACC-83 cells were seeded in the medium pore with DRG around them. Outgrowth of neuronal processes was observed. Then DRG was cultured in the conditioned medium of SACC-83, with the groups of conditioned medium of MC3T3-E1 and HGF, the group of cell lysis buffer, the groups of serum-free medium and serum-plus medium as the controls. Outgrowth of neuronal processes was also recorded and compared with control groups. In the co-culture model of tumor and neuronal tissue, SACC-83 cells produced a suitable microenvironment in which neuronal processes remarkably grow. Neuronal processes of most DRG displayed growth tendency toward SACC. The group of conditioned medium from SACC-83 manifested obvious promotive effects on DRG. Co-culture model of tumor and neuronal tissue was successfully constructed, with which the promotive effects of tumor on outgrowth of neuronal processes could be observed. So hypothesized that SACC could secrete some neurotrophic factors to guide peripheral nerves gemmating and to trigger the cascade of the neural invasion in succession.

p-Coumaric Acid Attenuates UVB-Induced Release of Stratifin from Keratinocytes and Indirectly Regulates Matrix Metalloproteinase 1 Release from Fibroblasts

PubMed Central

Seok, Jin Kyung

2015-01-01

Ultraviolet (UV) radiation-induced loss of dermal extracellular matrix is associated with skin photoaging. Recent studies demonstrated that keratinocyte-releasable stratifin (SFN) plays a critical role in skin collagen metabolism by inducing matrix metalloproteinase 1 (MMP1) expression in target fibroblasts. In the present study, we examined whether SFN released from UVB-irradiated epidermal keratinocytes increases MMP1 release from dermal fibroblasts, and whether these events are affected by p-coumaric acid (p-CA), a natural phenolic compound with UVB-shielding and antioxidant properties. HaCaT cells were exposed to UVB in the absence and presence of p-CA, and the conditioned medium was used to stimulate fibroblasts in medium transfer experiments. The cells and media were analyzed to determine the expressions/releases of SFN and MMP1. UVB exposure increased SFN release from keratinocytes into the medium. The conditioned medium of UVB-irradiated keratinocytes increased MMP1 release from fibroblasts. The depletion of SFN using a siRNA rendered the conditioned medium of UVB-irradiated keratinocytes ineffective at stimulating fibroblasts to release MMP1. p-CA mitigated UVB-induced SFN expression in keratinocytes, and attenuated the MMP1 release by fibroblasts in medium transfer experiments. In conclusion, the present study demonstrated that the use of UV absorbers such as p-CA would reduce UV-induced SFN-centered signaling events involved in skin photoaging. PMID:25954129

p-Coumaric Acid Attenuates UVB-Induced Release of Stratifin from Keratinocytes and Indirectly Regulates Matrix Metalloproteinase 1 Release from Fibroblasts.

PubMed

Seok, Jin Kyung; Boo, Yong Chool

2015-05-01

Ultraviolet (UV) radiation-induced loss of dermal extracellular matrix is associated with skin photoaging. Recent studies demonstrated that keratinocyte-releasable stratifin (SFN) plays a critical role in skin collagen metabolism by inducing matrix metalloproteinase 1 (MMP1) expression in target fibroblasts. In the present study, we examined whether SFN released from UVB-irradiated epidermal keratinocytes increases MMP1 release from dermal fibroblasts, and whether these events are affected by p-coumaric acid (p-CA), a natural phenolic compound with UVB-shielding and antioxidant properties. HaCaT cells were exposed to UVB in the absence and presence of p-CA, and the conditioned medium was used to stimulate fibroblasts in medium transfer experiments. The cells and media were analyzed to determine the expressions/releases of SFN and MMP1. UVB exposure increased SFN release from keratinocytes into the medium. The conditioned medium of UVB-irradiated keratinocytes increased MMP1 release from fibroblasts. The depletion of SFN using a siRNA rendered the conditioned medium of UVB-irradiated keratinocytes ineffective at stimulating fibroblasts to release MMP1. p-CA mitigated UVB-induced SFN expression in keratinocytes, and attenuated the MMP1 release by fibroblasts in medium transfer experiments. In conclusion, the present study demonstrated that the use of UV absorbers such as p-CA would reduce UV-induced SFN-centered signaling events involved in skin photoaging.

Coconut as a Medium for the Experimental Production of Aflatoxin

PubMed Central

Arseculeratne, S. N.; De Silva, L. M.; Wijesundera, S.; Bandunatha, C. H. S. R.

1969-01-01

Fresh, grated coconut has been found to be an excellent medium for aflatoxin production by Aspergillus flavus. Under optimal conditions, yields of 8 mg of total aflatoxin per g of substrate were obtained. Continuous agitation of the growth medium under moist conditions at 24 C produced highest yields. Aflatoxin was assayed both biologically and chromatographically. The aflatoxin content of cultures varied biphasically with the duration of incubation. It is suggested that this pattern could result from the sequential operation of factors promoting aflatoxin formation on the one hand and a detoxifying mechanism on the other. Images PMID:5803632

32 CFR 327.6 - Collecting personal information

Code of Federal Regulations, 2012 CFR

2012-07-01

... records, or similar information. (c) Collecting social security numbers (SSNs). (1) It is unlawful for... inclusion in a system of records, a Privacy Act Statement is required regardless of the medium used to...

32 CFR 327.6 - Collecting personal information.

Code of Federal Regulations, 2014 CFR

2014-07-01

... records, or similar information. (c) Collecting social security numbers (SSNs). (1) It is unlawful for... inclusion in a system of records, a Privacy Act Statement is required regardless of the medium used to...

32 CFR 327.6 - Collecting personal information

Code of Federal Regulations, 2010 CFR

2010-07-01

... records, or similar information. (c) Collecting social security numbers (SSNs). (1) It is unlawful for... inclusion in a system of records, a Privacy Act Statement is required regardless of the medium used to...

32 CFR 327.6 - Collecting personal information

Code of Federal Regulations, 2011 CFR

2011-07-01

... records, or similar information. (c) Collecting social security numbers (SSNs). (1) It is unlawful for... inclusion in a system of records, a Privacy Act Statement is required regardless of the medium used to...

32 CFR 327.6 - Collecting personal information.

Code of Federal Regulations, 2013 CFR

2013-07-01

... records, or similar information. (c) Collecting social security numbers (SSNs). (1) It is unlawful for... inclusion in a system of records, a Privacy Act Statement is required regardless of the medium used to...

Spoilage bacteria of fresh broiler chicken carcasses.

PubMed

Russell, S M; Fletcher, D L; Cox, N A

1995-12-01

Studies were conducted to identify the bacteria responsible for spoilage of fresh broiler chicken carcasses and to characterize the off-odors these bacteria produce. Broiler carcasses were collected from processing plants in the northeast Georgia area, the southeastern U.S., Arkansas, California, and North Carolina. The carcasses were allowed to spoil under controlled conditions at 3 C and spoilage bacteria were isolated. Each spoilage bacterium was separately inoculated into a sterile chicken skin medium, incubated at 25 C for 48 h, and subjectively evaluated for odor. The bacteria isolated from spoiled carcasses that consistently produced off-odors in the chicken skin medium, regardless of the geographical location from which the chickens were obtained, were Shewanella putrefaciens A, B, and D, Pseudomonas fluorescens A, B, and D, and Pseudomonas fragi. These bacteria produced off-odors that resembled "sulfur", "dishrag", "ammonia", "wet dog", "skunk", "dirty socks", "rancid fish", "unspecified bad odor", or a sweet smell resembling "canned corn". Odors produced by the spoilage bacteria were varied; however, odors most associated with spoiled poultry, such as "dishraggy" odors, were produced by the bacteria that were most consistently isolated, such as S. putrefaciens and the pseudomonads.

High Frequency High Spectral Resolution Focal Plane Arrays for AtLAST

NASA Astrophysics Data System (ADS)

Baryshev, Andrey

2018-01-01

Large collecting area single dish telescope such as ATLAST will be especially effective for medium (R 1000) and high (R 50000) spectral resolution observations. Large focal plane array is a natural solution to increase mapping speed. For medium resolution direct detectors with filter banks (KIDs) and or heterodyne technology can be employed. We will analyze performance limits of comparable KID and SIS focal plane array taking into account quantum limit and high background condition of terrestrial observing site. For large heterodyne focal plane arrays, a high current density AlN junctions open possibility of large instantaneous bandwidth >40%. This and possible multi frequency band FPSs presents a practical challenge for spatial sampling and scanning strategies. We will discuss phase array feeds as a possible solution, including a modular back-end system, which can be shared between KID and SIS based FPA. Finally we will discuss achievable sensitivities and pixel co unts for a high frequency (>500 GHz) FPAs and address main technical challenges: LO distribution, wire counts, bias line multiplexing, and monolithic vs. discrete mixer component integration.

Cytotoxic and hemolytic effects of Tritrichomonas foetus on mammalian cells.

PubMed Central

Burgess, D E; Knoblock, K F; Daugherty, T; Robertson, N P

1990-01-01

Geographically distinct lines of Tritrichomonas foetus were assayed for their ability to cause cytotoxicity in nucleated mammalian cells and lysis of bovine erythrocytes. T. foetus was highly cytotoxic toward a human cervical cell line (HeLa) and early bovine lymphosarcoma (BL-3) but displayed low levels of cytotoxicity against African green monkey kidney (Vero) cells. In addition to variation in the extent of cytotoxicity toward different targets, differences in the levels of cytotoxicity in the same nucleated target occurred with different parasite lines. Whole T. foetus, unfractionated whole-cell extracts, and parasite-conditioned medium (RPMI 1640 without serum) all caused lysis of bovine erythrocytes. Lytic activity in the conditioned medium was substantially reduced by repeated freezing and thawing or heating to 90 degrees C for 30 min. Damage of mammalian target cells by live T. foetus could be reduced by the presence of protease inhibitors; however, such inhibitors did not diminish the lytic effects of conditioned medium. These results suggested that proteolytic enzymes were necessary for the lytic mechanism of the live parasites but were not required once lytic factors were released into the parasite-conditioned medium. They further suggested that the lytic molecules were either proteins or had proteinaceous components. Images PMID:2228233

Race Does Not Predict Melanocyte Heterogeneous Responses to Dermal Fibroblast-Derived Mediators

PubMed Central

Sirimahachaiyakul, Pornthep; Sood, Ravi F.; Muffley, Lara A.; Seaton, Max; Lin, Cheng-Ta; Qiao, Liang; Armaly, Jeffrey S.; Hocking, Anne M.; Gibran, Nicole S.

2015-01-01

Introduction Abnormal pigmentation following cutaneous injury causes significant patient distress and represents a barrier to recovery. Wound depth and patient characteristics influence scar pigmentation. However, we know little about the pathophysiology leading to hyperpigmentation in healed shallow wounds and hypopigmentation in deep dermal wound scars. We sought to determine whether dermal fibroblast signaling influences melanocyte responses. Methods and Materials Epidermal melanocytes from three Caucasians and three African-Americans were genotyped for single nucleotide polymorphisms (SNPs) across the entire genome. Melanocyte genetic profiles were determined using principal component analysis. We assessed melanocyte phenotype and gene expression in response to dermal fibroblast-conditioned medium and determined potential mesenchymal mediators by proteome profiling the fibroblast-conditioned medium. Results Six melanocyte samples demonstrated significant variability in phenotype and gene expression at baseline and in response to fibroblast-conditioned medium. Genetic profiling for SNPs in receptors for 13 identified soluble fibroblast-secreted mediators demonstrated considerable heterogeneity, potentially explaining the variable melanocyte responses to fibroblast-conditioned medium. Discussion Our data suggest that melanocytes respond to dermal fibroblast-derived mediators independent of keratinocytes and raise the possibility that mesenchymal-epidermal interactions influence skin pigmentation during cutaneous scarring. PMID:26418010

Methodological considerations for implementation of lymphocyte subset analysis in a clinical reference laboratory.

PubMed

Muirhead, K A; Wallace, P K; Schmitt, T C; Frescatore, R L; Franco, J A; Horan, P K

1986-01-01

As the diagnostic utility of lymphocyte subset analysis has been recognized in the clinical research laboratory, a wide variety of reagents and cell preparation, staining and analysis methods have also been described. Methods that are perfectly suitable for analysis of smaller sample numbers in the biological or clinical research setting are not always appropriate and/or applicable in the setting of a high volume clinical reference laboratory. We describe here some of the specific considerations involved in choosing a method for flow cytometric analysis which minimizes sample preparation and data analysis time while maximizing sample stability, viability, and reproducibility. Monoclonal T- and B-cell reagents from three manufacturers were found to give equivalent results for a reference population of healthy individuals. This was true whether direct or indirect immunofluorescence staining was used and whether cells were prepared by Ficoll-Hypaque fractionation (FH) or by lysis of whole blood. When B cells were enumerated using a polyclonal anti-immunoglobulin reagent, less cytophilic immunoglobulin staining was present after lysis than after FH preparation. However, both preparation methods required additional incubation at 37 degrees C to obtain results concordant with monoclonal B-cell reagents. Standard reagents were chosen on the basis of maximum positive/negative separation and the availability of appropriate negative controls. The effects of collection medium and storage conditions on sample stability and reproducibility of subset analysis were also assessed. Specimens collected in heparin and stored at room temperature in buffered medium gave reproducible results for 3 days after specimen collection, using either FH or lysis as the preparation method. General strategies for instrument optimization, quality control, and biohazard containment are also discussed.

Isolation and in vitro culture of trypanosomes from Leptodactylus ocellatus from the Atlantic Forest in a new experimental culture medium.

PubMed

Lemos, M; Souza, C S F; da Costa, S C Gonçalves; Souto-Padrón, T; D'Agosto, M

2013-02-01

The purpose of this study was to verify the in vitro development of Trypanosoma sp. isolated from Leptodactylus ocellatus frogs under a new protocol using a biphasic medium composed of Novy, McNeal, and Nicolle (NNN) blood agar medium as a solid phase and liver infusion, brain heart infusion, and tryptose (LIBHIT) medium as a liquid phase. Blood forms, collected by cardiac puncture or after the maceration of different organs, were inoculated in culture tubes containing the biphasic medium composed by NNN and LIBHIT. Trypanosomes were observed 4 days postinoculation; most bloodstream trypomastigotes had differentiated into epimastigotes and amastigotes by this time. Trypomastigotes were again observed in older cultures (7 days). Parasites were successfully subcultured for 8 mo in this medium and successfully cryopreserved. The present study provides a new protocol medium for the isolation and culture of anuran trypanosomes.

Starch and lipid accumulation in eight strains of six Chlorella species under comparatively high light intensity and aeration culture conditions.

PubMed

Takeshita, Tsuyoshi; Ota, Shuhei; Yamazaki, Tomokazu; Hirata, Aiko; Zachleder, Vilém; Kawano, Shigeyuki

2014-04-01

The microalgae family Chlorella species are known to accumulate starch and lipids. Although nitrogen or phosphorous deficiencies promote starch and lipids formation in many microalgae, these deficiencies also limit their growth and productivity. Therefore, the Chlorellaceae strains were attempted to increase starch and lipids productivity under high-light-intensity conditions (600-μmol photons m(-2)s(-1)). The 12:12-h light-dark (LD) cycle conditions elicited more stable growth than the continuous light (LL) conditions, whereas the starch and lipids yields increased in LL conditions. The amount of starch and lipids per cell increased in Chlorella viscosa and Chlorella vulgaris in sulfur-deficient medium, and long-chain fatty acids with 20 or more carbon atoms accumulated in cells grown in sulfur-deficient medium. Accumulation of starch and lipids was investigated in eight strains. The accumulation was strain-dependent, and varied according to the medium and light conditions. Five of the eight Chlorella strains exhibited similar accumulation patterns. Copyright © 2014 Elsevier Ltd. All rights reserved.

An evaluation of clinical performance of FTA cards for HPV 16/18 detection using cobas 4800 HPV Test compared to dry swab and liquid medium.

PubMed

Dong, Li; Lin, Chunqing; Li, Li; Wang, Margaret; Cui, Jianfeng; Feng, Ruimei; Liu, Bin; Wu, Zeni; Lian, Jia; Liao, Guangdong; Chen, Wen; Qiao, Youlin

2017-09-01

Effective dry storage and transport media as an alternative to conventional liquid-based medium would facilitate the accessibility of women in the low-resource settings to human papillomavirus (HPV)- based cervical cancer screening. To evaluate analytical and clinical performance of indicating FTA™ Elute Cartridge (FTA card) for the detection of HPV16/18 and cervical precancerous lesions and cancer compared to dry swab and liquid medium. Ninety patients with abnormal cytology and/or HPV infection were included for analysis. Three specimens of cervical exfoliated cells from each woman were randomly collected by FTA card, dry swab or liquid-based medium prior to colposcopy examination. The subsequent HPV DNA tests were performed on cobas 4800 HPV platform. High-risk HPV (hrHPV) positivity rate was 63.3%, 62.2% and 65.6% for samples collected by FTA card, dry swab and liquid medium, respectively. The overall agreements and kappa values for the detection of hrHPV, HPV 16 and HPV 18 between FTA card and liquid-based medium were 88.9% (κ=0.76), 97.8% (κ=0.94) and 100% (κ=1.0),respectively; between FTA card and dry swab were 92.1% (κ=0.83), 94.5% (κ=0.87) and 100% (κ=1.0), respectively. The performances of hrHPV tested by FTA card, dry swab, and liquid-based medium for detecting CIN2+ were comparable in terms of the sensitivity and specificity. The specificity of detection of CIN2+ by HPV16/18 increased by approximately 40% compared to hrHPV for any medium albeit at cost of a moderate loss of sensitivity. Dry medium might offer an alternative to conventional liquid-based medium in the HPV-based cervical cancer screening program especially in low-resource settings but still needs further evaluation. Copyright © 2017. Published by Elsevier B.V.

HYDROGENATION OF 4-OXOISOPHORONE OVER PD/AL2/O2 CATALYST UNDER SUPERCRITICAL CO2 MEDIUM

EPA Science Inventory

Hydrogenation of 4-oxoisophorone has been studied over 1% Pd/Al2O3 impregnated catalyst in supercritical CO2 medium at different reaction conditions. The effect of temperature, pressure and reaction medium on the conversion and product selectivity is discussed. Phase behavior stu...

Density-dependent regulation of growth of BSC-1 cells in cell culture: growth inhibitors formed by the cells.

PubMed Central

Holley, R W; Armour, R; Baldwin, J H

1978-01-01

Inhibitors formed by a monkey epithelial cell line, BSC-1, play an important role in limiting growth at high cell densities. At least three inhibitors are formed: lactic acid, ammonia, and an unidentified inhibitor that may be an unstable protein. The unidentified inhibitor is destroyed by shaking the conditioned medium, by bubbling gas through the medium, or by heating or storing the medium in the absence of cells. The concentrations of lactic acid and ammonia that accumulate in conditioned medium inhibit growth when added to fresh medium. These results, together with earlier studies, indicate that density-dependent regulation of growth of BSC-1 cells results from the combined effects of (a) inhibitors formed by the cells, (b) decreased availability of receptor sites for serum growth factors as the cells become crowded, and (c) limiting concentrations of low molecular weight nutrients in the medium. In contrast, density-dependent regulation of growth in 3T3 mouse embryo fibroblasts results almost entirely from inactivation of serum factors. PMID:273914

Evaluation of the Universal Viral Transport system for long-term storage of virus specimens for microbial forensics.

PubMed

Hosokawa-Muto, Junji; Fujinami, Yoshihito; Mizuno, Natsuko

2015-08-01

Forensic microbial specimens, including bacteria and viruses, are collected at biocrime and bioterrorism scenes. Although it is preferable that the pathogens in these samples are alive and kept in a steady state, the samples may be stored for prolonged periods before analysis. Therefore, it is important to understand the effects of storage conditions on the pathogens contained within such samples. To evaluate the capacity to preserve viable virus and the viral genome, influenza virus was added to the transport medium of the Universal Viral Transport system and stored for over 3 months at various temperatures, after which virus titrations and quantitative analysis of the influenza hemagglutinin gene were performed. Although viable viruses became undetectable 29 days after the medium was stored at room temperature, viruses in the medium stored at 4°C were viable even after 99 days. A quantitative PCR analysis indicated that the hemagglutinin gene was maintained for 99 days at both 4°C and room temperature. Therefore, long-term storage at 4°C has little effect on viable virus and viral genes, so the Universal Viral Transport system can be useful for microbial forensics. This study provides important information for the handling of forensic virus specimens. Copyright © 2015 Elsevier Ltd and Faculty of Forensic and Legal Medicine. All rights reserved.

Hypoxic Conditioned Medium From Human Adipose-Derived Stem Cells Promotes Mouse Liver Regeneration Through JAK/STAT3 Signaling

PubMed Central

Lee, Sang Chul; Jeong, Hye Jin; Lee, Sang Kuon

2016-01-01

Adipose-derived stem cells (ASCs) mainly exert their function by secreting materials that are collectively termed the secretome. Despite recent attention to the secretome as an alternative to stem cell therapy, the culture conditions for generating optimal secretome contents have not been determined. Therefore, we investigated the role of hypoxic-conditioned media (HCM) from ASCs. Normoxic-conditioned media (NCM) and HCM were obtained after culturing ASCs in 20% O2 or 1% O2 for 24 hours, respectively. Subsequently, partially hepatectomized mice were infused with saline, control medium, NCM, or HCM, and then sera and liver specimens were obtained for analyses. Hypoxia (1% O2) significantly increased mRNA expression of mediators from ASCs, including interleukin-6 (IL-6), tumor necrosis factor α (TNF-α), hepatocyte growth factor (HGF), and vascular endothelial growth factor (VEGF). HCM infusion significantly increased the number of Ki67-positive cells in the liver (p < .05). HCM infusion significantly increased phospho-signal transducer and activator of transcription 3 (STAT3) and decreased suppressor of cytokine signaling 3 (SOCS3) expression in the liver (p < .05). To determine the role of IL-6 in liver regeneration, we then performed IL-6 RNA interference study. Conditioned media (CM) obtained from ASCs, which were transfected with either siIL-6 or siControl, were administered to partially hepatectomized mice. The siIL-6 CM groups exhibited lower liver proliferation (Ki67-positive cells) and markers of regeneration (protein expression of proliferating cell nuclear antigen, p-STAT3, HGF, and VEGF and liver weights) than the siControl CM groups (p < .05). Taken together, hypoxic preconditioning of ASCs increased expression of mediators promoting anti-inflammatory and regenerative responses. The liver regenerative effects of HCM appear to be mediated by persistent and uninhibited expression of STAT3 in the liver, which results from decreased expression of SOCS3. Significance In this study, it was found that treatment with the medium from hypoxic-preconditioned adipose-derived stem cells (ASCs) increased the viability of hepatotoxic hepatocytes and enhance liver regeneration in partially hepatectomized mice. In addition, the researchers first revealed that the hepatoprotective effects of hypoxic-conditioned media are mediated by persistent and uninhibited expression of signal transducer and activator of transcription 3 in the liver, which result from a decreased expression of suppressor of cytokine signaling 3. Therefore, the hypoxic preconditioning of ASCs is expected to play a crucial role in regenerative medicine by optimizing the production of a highly effective secretome from ASCs. PMID:27102647

Cytotoxicity evaluation of a copaiba oil-based root canal sealer compared to three commonly used sealers in endodontics

PubMed Central

Garrido, Angela Delfina Bittencourt; de Cara, Sueli Patricia Harumi Miyagi; Marques, Marcia Martins; Sponchiado, Emílio Carlos; Garcia, Lucas da Fonseca Roberti; de Sousa-Neto, Manoel Damião

2015-01-01

Background: The constant development of new root canal sealers has allowed the solution of a large number of clinical cases in endodontics, however, cytotoxicity of such sealers must be tested before their validation as filling materials. The aim of this study was to evaluate the cytotoxic effect of a new Copaiba oil-based root canal sealer (Biosealer [BS]) on osteoblast-like Osteo-1 cells. Materials and Methods: The experimental groups were formed according to the culture medium conditioned with the tested sealers, as follows: Control group (CG) (culture medium without conditioning); Sealer 26 (S26) - culture medium + S26; Endofill (EF) - culture medium + EF; AH Plus (AHP) - culture medium + AHP; and BS - culture medium + BS (Copaiba oil-based sealer). The conditioned culture medium was placed in contact with 2 × 104 cells cultivated on 60 mm diameter Petri dishes for 24 h. Then, hemocytometer count was performed to evaluate cellular viability, using Trypan Blue assay. The normal distribution of data was tested by the Kolmogorov-Smirnov test and the values obtained for cellular viability were statistically analyzed (1-way ANOVA, Tukey's test - P < 0.05), with a significance level of 5%. Results: S26, EF and AHP presented decreased cellular viability considerably, with statistical significance compared with CG (P < 0.05). BS maintained cellular viability similar to CG (P > 0.05). Conclusion: The Copaiba oil-based root canal sealer presented promising results in terms of cytotoxicity which indicated its usefulness as a root canal sealer. PMID:25878676

Effects of medium components and culture conditions on mycelial biomass and the production of bioactive ingredients in submerged culture of Xylaria nigripes (Ascomycetes), a Chinese medicinal fungus.

PubMed

Chen, Jian-Zhi; Lo, Hui-Chen; Lin, Fang-Yi; Chang, Shih-Liang; Hsieh, Changwei; Liang, Zeng-Chin; Ho, Wai-Jane; Hsu, Tai-Hao

2014-01-01

The optimal culture conditions were investigated to maximize the production of mycelial biomass and bioactive ingredients in submerged cultivation of Xylaria nigripes, a Chinese medicinal fungus. The one-factor-at-a-time method was used to explore the effects of medium components, including carbon, nitrogen, mineral sources, and initial pH of the medium and environmental factors, such as culture temperature and rotation speed, on mycelial growth and production of bioactive ingredients. The results indicated that the optimal culture temperature and rotation speed were 25°C and 100 rpm in a medium with 20 g fructose, 6 g yeast extract, and 2 g magnesiun sulfate heptahydrate as carbon, nitrogen, and mineral sources, respectively, in 1 L distilled water with an initial medium pH of 5.5. With optimal medium components and conditions of cultivation, the maximal production of mycelial biomass was 6.64 ± 0.88 g/L, with maximal production of bioactive ingredients such as extracellular polysaccharides (2.36 ± 0.18 mg/mL), intracellular polysaccharides (2.38 ± 0.07 mg/g), adenosine (43.27 ± 2.37 mg/g), total polyphenols (36.57 ± 1.36 mg/g), and triterpenoids (31.29 ± 1.17 mg/g) in a shake flask culture. These results suggest that different bioactive ingredients including intracellular polysaccharides, adenosine, total polyphenols and triterpenoids in mycelia and extracellular polysaccharides in broth can be obtained from one simple medium for submerged cultivation of X. nigripes.

Time of travel and dispersion in a selected reach of Roberts Creek, Clayton County, Iowa

USGS Publications Warehouse

Kolpin, D.W.; Kalkhoff, S.J.

1992-01-01

Time of travel was determined by dye tracing, using rhodamine WT as the tracer. One dyeinjection site and three sampling sites were used to measure time of travel. Two dye-tracing tests were conducted at discharges having flow-duration values of 48 and 80 percent. The discharges at the time of the two dye-tracing tests approximated medium- and low-flow conditions. The average stream velocity in the study area was 0.23 foot per second during medium-flow conditions, March 20 to 22,1990, and 0.07 foot per second during low-flow conditions, April 30 to May 12, 1990. The injected dye dispersed in a plume that lasted about 18 hours during medium flow and about 64 hours during low flow at the downstream site.

Numerical model for the uptake of groundwater contaminants by phreatophytes

USGS Publications Warehouse

Widdowson, M.A.; El-Sayed, A.; Landmeyer, J.E.

2008-01-01

Conventional solute transport models do not adequately account for the effects of phreatophytic plant systems on contaminant concentrations in shallow groundwater systems. A numerical model was developed and tested to simulate threedimensional reactive solute transport in a heterogeneous porous medium. Advective-dispersive transport is coupled to biodegradation, sorption, and plantbased attenuation processes including plant uptake and sorption by plant roots. The latter effects are a function of the physical-chemical properties of the individual solutes and plant species. Models for plant uptake were tested and evaluated using the experimental data collected at a field site comprised of hybrid poplar trees. A non-linear equilibrium isotherm model best represented site conditions.

Hydrogen production from microbial strains

DOEpatents

Harwood, Caroline S; Rey, Federico E

2012-09-18

The present invention is directed to a method of screening microbe strains capable of generating hydrogen. This method involves inoculating one or more microbes in a sample containing cell culture medium to form an inoculated culture medium. The inoculated culture medium is then incubated under hydrogen producing conditions. Once incubating causes the inoculated culture medium to produce hydrogen, microbes in the culture medium are identified as candidate microbe strains capable of generating hydrogen. Methods of producing hydrogen using one or more of the microbial strains identified as well as the hydrogen producing strains themselves are also disclosed.

Quantification of the extracellular matrix of the Listeria monocytogenes biofilms of different phylogenic lineages with optimization of culture conditions.

PubMed

Combrouse, T; Sadovskaya, I; Faille, C; Kol, O; Guérardel, Y; Midelet-Bourdin, G

2013-04-01

The purpose of this study was to quantify the extracellular matrix of Listeria monocytogenes biofilm. A preliminary study was carried out to establish a relationship between phylogenetic lineage of 27 strains and their ability to form biofilm in various conditions. Biofilm formation on microtitre plates of 27 strains of L. monocytogenes belonging to lineages I or II was evaluated in different conditions [two temperatures (37 and 22°C) and two media (tryptone soy broth yeast extract medium (TSBYE) and MCDB 202 defined medium)] using crystal violet assay. Lineage II strains produced significantly more biofilm than lineage I strains. In microtitre plates assay, biofilm quantities were greater in MCDB 202 vs TSBYE medium [confirmed by scanning electron microscopy (SEM) analysis] and at 37 vs 22°C. Cultivable bacteria from biofilm population on Petri dishes were enumerated in greater quantities in TSBYE than in MCDB 202 medium. The SEM investigation established that L. monocytogenes biofilms produce extracellular matrix in both media at 37°C. The amount of exopolymers in the extracellular matrix and the pH values were significantly higher in TSBYE than in MCDB 202 medium. The exception was the ScottA strain that presented similar pH values and exopolymer contents in both media. Proteins were the most abundant exopolymer components, followed by DNA and polysaccharides. The interpretation of results of biofilm quantification was depending on the growth conditions, the viability of the bacteria and the analysis method. The quantities of proteins, DNA and polysaccharides were different according to the strains and the medium. This study screened the potential of a wide panel of L. monocytogenes strains to synthesize exopolymers in biofilm growing condition. The characterization of L. monocytogenes biofilm composition may help to develop new strategies to prevent the formation of biofilms and to remove the biofilms. © 2013 The Society for Applied Microbiology.

Platelet-rich plasma can replace fetal bovine serum in human meniscus cell cultures.

PubMed

Gonzales, Veronica K; de Mulder, Eric L W; de Boer, Trix; Hannink, Gerjon; van Tienen, Tony G; van Heerde, Waander L; Buma, Pieter

2013-11-01

Concerns over fetal bovine serum (FBS) limit the clinical application of cultured tissue-engineered constructs. Therefore, we investigated if platelet-rich plasma (PRP) can fully replace FBS for meniscus tissue engineering purposes. Human PRP and platelet-poor plasma (PPP) were isolated from three healthy adult donors. Human meniscal fibrochondrocytes (MFCs) were isolated from resected tissue after a partial meniscectomy on a young patient. Passage-4 MFCs were cultured in monolayer for 24 h, and 3 and 7 days. Six different culture media were used containing different amounts of either PRP or PPP and compared to a medium containing 10% FBS. dsDNA was quantified, and gene expression levels of collagen types I and II and aggrecan were measured at different time points with quantitative polymerase chain reaction in the cultured MFCs. After 7 days, the dsDNA quantity was significantly higher in MFCs cultured in 10% and 20% PRP compared to the other PRP and PPP conditions, but equal to 10% FBS. Collagen type I expression was lower in MFCs cultured with medium containing 5% PRP, 10% and 20% PPP compared to FBS. When medium with 10% PRP or 20% PRP was used, expressions were not significantly different from medium containing 10% FBS. Collagen type II expression was absent in all medium conditions. Aggrecan expression did not show differences between the different media used. However, after 7 days a higher aggrecan expression was measured in most culture conditions, except for 5% PRP, which was similar compared to FBS. Statistical significance was found between donors at various time points in DNA quantification and gene expression, but the same donors were not statistically different in all conditions. At 7 days cell cultured with 10% PRP and 20% PRP showed a higher density, with large areas of clusters, compared to other conditions. In an MFC culture medium, FBS can be replaced by 10% PRP or 20% PRP without altering proliferation and gene expression of human MFCs.

Statistical optimization of medium composition and culture condition for the production of recombinant anti-lipopolysaccharide factor of Eriocheir sinensis in Escherichia coli

NASA Astrophysics Data System (ADS)

Jiang, Shan; Liu, Mei; Wang, Baojie; Jiang, Keyong; Wang, Lei

2011-11-01

Anti-lipopolysaccharide factors (ALFs) are important antimicrobial peptides that are isolated from some aquatic species. In a previous study, we isolated ALF genes from Chinese mitten crab, Eriocheir sinensis. In this study, we optimized the production of a recombinant ALF by expressing E. sinensis ALF genes in Escherichia coli maintained in shake-flasks. In particular, we focused on optimization of both the medium composition and the culture condition. Various medium components were analyzed by the Plackett-Burman design, and two significant screened factors, (NH4)2SO4 and KH2PO4, were further optimized via the central composite design (CCD). Based on the CCD analysis, we investigated the induction start-up time, the isopropylthio-D-galactoside (IPTG) concentration, the post-induction time, and the temperature by response surface methodology. We found that the highest level of ALF fusion protein was achieved in the medium containing 1.89 g/L (NH4)2SO4 and 3.18 g/L KH2PO4, with a cell optical density of 0.8 at 600 nm before induction, an IPTG concentration of 0.5 mmol/L, a post-induction temperature of 32.7°C, and a post-induction time of 4 h. Applying the whole optimization strategy using all optimal factors improved the target protein content from 6.1% (without optimization) to 13.2%. We further applied the optimized medium and conditions in high cell density cultivation, and determined that the soluble target protein constituted 10.5% of the total protein. Our identification of the economic medium composition, optimal culture conditions, and details of the fermentation process should facilitate the potential application of ALF for further research.

Analytical results for a conditional phase shift between single-photon pulses in a nonlocal nonlinear medium

NASA Astrophysics Data System (ADS)

Viswanathan, Balakrishnan; Gea-Banacloche, Julio

2017-04-01

We analyze a recent scheme proposed by Xia et al. to induce a conditional phase shift between two single-photon pulses by having them propagate at different speeds through a nonlinear medium with a nonlocal response. We have obtained an analytical solution for the case they considered, which supports their claim that a π phase shift with unit fidelity is possible in principle. We discuss the conditions that have to be met and the challenges and opportunities that this might present to the realization of a single-photon conditional phase gate.

Medium- and long-term storage of the Pycnanthemum (Mountain mint) germplasm collection

USDA-ARS?s Scientific Manuscript database

The US collection of mountain mint (Pycnanthemum Michx.) is held at the USDA-ARS National Clonal Germplasm Repository (NCGR) in Corvallis, OR as seed, potted plants and tissue cultures and a long-term storage collection is preserved at the USDA-ARS National Center for Genetic Resources Preservation ...

The in vitro impact of toothpaste extracts on cell viability.

PubMed

Cvikl, Barbara; Lussi, Adrian; Gruber, Reinhard

2015-06-01

Toothpastes contain three main components: detergents, abrasives, and fluoride. Detergents, particularly sodium lauryl sulfate, have been proposed as components that enable toothpastes to produce cytotoxic effects in vitro. However, not all toothpastes contain sodium lauryl sulfate, and almost no studies have found an association between detergents and the in vitro cytotoxicity of toothpastes. The present study examined the in vitro cytotoxicity of nine commercially available toothpastes containing four different detergents. Toothpastes were diluted in serum-free medium, centrifuged, and filter sterilized. The half-lethal concentration of the toothpaste-conditioned medium (TCM) was calculated based on the formation of formazan by gingival fibroblasts, oral squamous cell carcinoma HSC-2 cells, and L929 cells. Cell proliferation was analyzed, and live-dead staining was performed, after exposure of cells to conditioned medium prepared with 1% toothpaste (1% TCM). It was found that toothpastes containing sodium lauryl sulfate and amine fluoride strongly inhibited cell viability with the half-lethal concentration being obtained with conditioned medium prepared with approximately 1% toothpaste (1% TCM). Toothpastes containing cocamidopropyl betaine and Steareth-20 showed higher half-lethal concentration values, with the half-lethal concentration being obtained with conditioned medium prepared with 10% (10% TCM) and 70% (70% TCM) toothpaste, respectively. Proliferation and live-dead data were consistent with the cell-viability analyses. These results demonstrate that the type of detergent in toothpastes can be associated with changes in in vitro cell toxicity. © 2015 Eur J Oral Sci.

Grain dust originating from organic and conventional farming as a potential source of biological agents causing respiratory diseases in farmers.

PubMed

Zukiewicz-Sobczak, Wioletta A; Cholewa, Grażyna; Krasowska, Ewelina; Chmielewska-Badora, Jolanta; Zwoliński, Jacek; Sobczak, Paweł

2013-12-01

Agricultural producers are exposed to a number of different health risks associated with their work environment. The objective of the study was to assess the degree of colonization by fungi in terms of quantity and in terms of variety of species the samples taken from the settled dust from combine threshing of rye cultivation from organic and conventional farms in the Province of Lublin. This paper is a preliminary quantitative assessment of the species of fungi colonizing the samples of settled dust collected during combine threshing from organic and conventional farms in the Province of Lublin. One of the stages of the project was the classification of biosafety BSL (biosafety level) of selected isolates and API ZYM tests to evaluate the potential ability of isolates to cause adverse health effects. To determine the concentration and composition of fungi in collected samples plate dilution method was used with two media: Malt Agar and Potato Dextrose Agar. MOST COMMONLY ISOLATED FUNGI IN SETTLED DUST SAMPLES COLLECTED DURING COMBINE THRESHING FROM ORGANIC FARMS, ON PDA MEDIUM WERE: Alternaria alternata and Aureobasidium pullulans. Cultures on MA medium were dominated by Alternaria alternata, Mycelia sterilia and Fusarium poae. In samples of dust from conventional crops, the predominant species was Alternaria alternata on PDA medium and on MA medium. The obtained results show a potential risk of people involved in agricultural work.

Effects of mineral trioxide aggregate, BiodentineTM and calcium hydroxide on viability, proliferation, migration and differentiation of stem cells from human exfoliated deciduous teeth.

PubMed

Araújo, Leandro Borges; Cosme-Silva, Leopoldo; Fernandes, Ana Paula; Oliveira, Thais Marchini de; Cavalcanti, Bruno das Neves; Gomes Filho, João Eduardo; Sakai, Vivien Thiemy

2018-02-01

The aim of the study was to evaluate the effects of the capping materials mineral trioxide aggregate (MTA), calcium hydroxide (CH) and BiodentineTM (BD) on stem cells from human exfoliated deciduous teeth (SHED) in vitro. SHED were cultured for 1 - 7 days in medium conditioned by incubation with MTA, BD or CH (1 mg/mL), and tested for viability (MTT assay) and proliferation (SRB assay). Also, the migration of serum-starved SHED towards conditioned media was assayed in companion plates, with 8 μm-pore-sized membranes, for 24 h. Gene expression of dentin matrix protein-1 (DMP-1) was evaluated by reverse-transcription polymerase chain reaction. Regular culture medium with 10% FBS (without conditioning) and culture medium supplemented with 20% FBS were used as controls. MTA, CH and BD conditioned media maintained cell viability and allowed continuous SHED proliferation, with CH conditioned medium causing the highest positive effect on proliferation at the end of the treatment period (compared with BD and MTA) (p<0.05). In contrast, we observed increased SHED migration towards BD and MTA conditioned media (compared with CH) (p<0.05). A greater amount of DMP-1 gene was expressed in MTA group compared with the other groups from day 7 up to day 21. Our results show that the three capping materials are biocompatible, maintain viability and stimulate proliferation, migration and differentiation in a key dental stem cell population.

Optimization of medium composition and cultural conditions for production of antifungal substances by Streptomyces platensis 3-10 and evaluation of its efficacy in suppression of clubroot disease of oilseed rape

USDA-ARS?s Scientific Manuscript database

This study was conducted to optimize the medium composition and cultural conditions for improving production of antifungal substances (AFS) by Streptomyces 3-10 and for enhancing its efficacy in suppression of clubroot disease of oilseed rape caused by Plasmodiophora brassicae. Results showed that t...

Glutathione transferase-M2-2 secreted from glioblastoma cell protects SH-SY5Y cells from aminochrome neurotoxicity.

PubMed

Cuevas, Carlos; Huenchuguala, Sandro; Muñoz, Patricia; Villa, Monica; Paris, Irmgard; Mannervik, Bengt; Segura-Aguilar, Juan

2015-04-01

U373MG cells are able to take up aminochrome that induces glutathione transferase M2-2 (GSTM2) expression in a concentration-dependent manner where 100 µM aminochrome increases GSTM2 expression by 2.1-fold (P < 0.001) at 3 h. The uptake of (3)H-aminochrome into U373MG cells was significantly reduced in the presence of 2 µM nomifensine (P < 0.001) 100 µM imipramine (P < 0.001) and 50 mM dopamine (P < 0.001). Interestingly, U373MG cells excrete GSTM2 into the conditioned medium and the excretion was significantly increased (2.7-fold; P < 0.001) when the cells were pretreated with 50 µM aminochrome for 3 h. The U373MG-conditioned medium containing GSTM2 protects SH-SY5Y cells incubated with 10 µM aminochrome. The significant protection provided by U373MG-conditioned medium in SH-SY5Y cells incubated with aminochrome was dependent on GSTM2 internalization into SH-SY5Y cells as evidenced by (i) uptake of (14)C-GSTM2 released from U373MG cells into SH-SY5Y cells, a process inhibited by anti-GSTM2 antiserum; (ii) lack of protection of U373MG-conditioned medium in the presence of anti-GSTM2 antiserum on SH-SY5Y cells treated with aminochrome; and (iii) lack of protection of conditioned medium from U373MGsiGST6 that expresses an siRNA directed against GSTM2 on SH-SY5Y cells treated with aminochrome. In conclusion, our results demonstrated that U373MG cells protect SH-SY5Y cells against aminochrome neurotoxicity by releasing GSTM2 into the conditioned medium and subsequent internalization of GSTM2 into SH-SY5Y cells. These results suggest a new mechanism of protection of dopaminergic neurons mediated by astrocytes by releasing GSTM2 into the intersynaptic space and subsequent internalization into dopaminergic neuron in order to protect these cells against aminochrome neurotoxicity.

77 FR 2697 - Proposed Information Collection; Comment Request; Annual Services Report

Federal Register 2010, 2011, 2012, 2013, 2014

2012-01-19

... and from a sample of small- and medium-sized businesses selected using a stratified sampling procedure... be canvassed when the sample is re-drawn, while nearly all of the small- and medium-sized firms from...); Educational Services (NAICS 61); Health Care and Social Assistance (NAICS 62); Arts, Entertainment, and...

Rural Press for Village Post-Literacy Literature.

ERIC Educational Resources Information Center

Sam-Kpakra, Robert H.; And Others

Three articles concerning the development of the rural press in Africa as a means of communicating and promoting literacy are included in this collection of reprints. "The Rural Press, Effective Rural Communication Medium" by Robert H. Sam-Kpakra describes this medium of communication by and for people living in areas with little or no…

Feasibility of Adopting English-Medium Instruction at Iranian Universities

ERIC Educational Resources Information Center

Ghorbani, Mohammad Reza; Alavi, Sahar Zahed

2014-01-01

This study was an attempt to explore the potential possibilities of implementing English as the medium of instruction (EMI) at Iranian universities. The sequential exploratory mixed methods design was used to collect the perspectives of both students and content-area lecturers at the state University of Bojnord through e-mail interviews and survey…

Unintended Results of Using Instructional Media: A Study of Second- and Third-Graders.

ERIC Educational Resources Information Center

Flanagan, Robin

Much of the research on classroom use of educational media has been hampered by difficulties in isolating a single element of the medium--television programming, for instance--that influences behavior in a reliable way. Still, each medium facilitates a particular type of learning environment, and the collective characteristics of those…

Self-Regulated Strategic Writing for Academic Studies in an English-Medium-Instruction Context

ERIC Educational Resources Information Center

Hu, Jingjing; Gao, Xuesong

2018-01-01

This study explored the processes of utilization of resources in secondary students' self-regulated strategic writing for academic studies in an English as medium of instruction context in Hong Kong. Drawing on multiple data sources collected through the observation of lessons, stimulated recall and semi-structured interviews, the study examined…

76 FR 74003 - Atlantic Highly Migratory Species; Adjustments to the Atlantic Bluefin Tuna General and Harpoon...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-11-30

... the extension dependent on weather conditions and availability of large medium and giant BFT to the... vessel. NMFS may increase or decrease the actual allowed daily retention limit of large medium and giant... NMFS implements an increase to the Harpoon category daily incidental retention limit of large medium...

Nanoparticle growth and surface chemistry changes in cell-conditioned culture medium.

PubMed

Kendall, Michaela; Hodges, Nikolas J; Whitwell, Harry; Tyrrell, Jess; Cangul, Hakan

2015-02-05

When biomolecules attach to engineered nanoparticle (ENP) surfaces, they confer the particles with a new biological identity. Physical format may also radically alter, changing ENP stability and agglomeration state within seconds. In order to measure which biomolecules are associated with early ENP growth, we studied ENPs in conditioned medium from A549 cell culture, using dynamic light scattering (DLS) and linear trap quadrupole electron transfer dissociation mass spectrometry. Two types of 100 nm polystyrene particles (one uncoated and one with an amine functionalized surface) were used to measure the influence of surface type. In identically prepared conditioned medium, agglomeration was visible in all samples after 1 h, but was variable, indicating inter-sample variability in secretion rates and extracellular medium conditions. In samples conditioned for 1 h or more, ENP agglomeration rates varied significantly. Agglomerate size measured by DLS was well correlated with surface sequestered peptide number for uncoated but not for amine coated polystyrene ENPs. Amine-coated ENPs grew much faster and into larger agglomerates associated with fewer sequestered peptides, but including significant sequestered lactose dehydrogenase. We conclude that interference with extracellular peptide balance and oxidoreductase activity via sequestration is worthy of further study, as increased oxidative stress via this new mechanism may be important for cell toxicity. © 2014 The Author(s) Published by the Royal Society. All rights reserved.

Growth of nutrient-replete Microcystis PCC 7806 cultures is inhibited by an extracellular signal produced by chlorotic cultures.

PubMed

Dagnino, Denise; de Abreu Meireles, Diogo; de Aquino Almeida, João Carlos

2006-01-01

The frequency of cyanobacterial blooms has been increasing all over the world. These blooms are often toxic and have become a serious health problem. The aim of this work was to search for population density control mechanisms that could inhibit the proliferation of the toxic bloom-forming genus Microcystis. Microcystis PCC 7806 cultured for long periods in liquid ASM-1 medium loses its characteristic green colour. When a medium of chlorotic cultures is added to a nutrient-replete culture, cell density increase is drastically reduced when compared with controls. Inhibition of cell proliferation occurs in Microcystis cultures from any growth stage and was not strain-specific, but other genera tested showed no response. Investigations on the mechanism of growth inhibition showed that cultures treated with the conditioned medium acquired a pale colour, with pigment concentration similar to that found in chlorotic cultures. Ultrastructural examination showed that the conditioned medium induced thylakoid membrane disorganization, typical of chlorotic cells, in nutrient-replete cultures. An active extract was obtained and investigations showed that activity was retained after heating and after addition of an apolar solvent. This indicates that activity of the conditioned medium from chlorotic cells results from non-protein, apolar compound(s).

Technique for restoration of mite (Acari) preparations in deteriorated Hoyer's medium.

PubMed

Jacinavicius, F C; Badari, J C; Ramirez, D G; Moraes, R H P; Onofrio, V C; Barros-Battesti, D M

2013-06-01

The Acari Collection of Instituto Butantan (IBSP), São Paulo, Brazil, includes many types and other identified mite specimens that were mounted in Hoyer's medium, mainly in the first part of last century. An effort to restore degraded preparations was initiated in 1996. In this process, an improved technique was developed, allowing the adequate cleaning of specimens mounted up to 50-70 years before. Types and other identified specimens of Trombidiformes (Harpirhynchidae and Trombiculidae), Sarcoptiformes (Acaridae, Atopomelidae, Listrophoridae, and Psoroptidae) and Mesostigmata (Dermanyssidae, Ixodorhynchidae, Laelapidae, Macronyssidae, and Spinturnicidae) deposited at IBSP Collection have been satisfactorily restored.

Method of treating contaminated HEPA filter media in pulp process

DOEpatents

Hu, Jian S.; Argyle, Mark D.; Demmer, Ricky L.; Mondok, Emilio P.

2003-07-29

A method for reducing contamination of HEPA filters with radioactive and/or hazardous materials is described. The method includes pre-processing of the filter for removing loose particles. Next, the filter medium is removed from the housing, and the housing is decontaminated. Finally, the filter medium is processed as pulp for removing contaminated particles by physical and/or chemical methods, including gravity, flotation, and dissolution of the particles. The decontaminated filter medium is then disposed of as non-RCRA waste; the particles are collected, stabilized, and disposed of according to well known methods of handling such materials; and the liquid medium in which the pulp was processed is recycled.

Harvesting microalgae cultures with superabsorbent polymers: desulfurization of Chlamydomonas reinhardtii for hydrogen production.

PubMed

Martín del Campo, Julia S; Patiño, Rodrigo

2013-12-01

It is presented in this work a new methodology to harvest fresh water microalgae cultures by extracting the culture medium with superabsorbent polymers (SAPs). The microalgae Chlamydomonas reinhardtii were grown in the Sueoka culture medium, harvested with polyacrylic SAPs and re-suspended in the culture medium tris-acetate-potassium without sulfur (TAP-S) to generate hydrogen (H2 ) under anoxic conditions. The H2 production as an alternative fuel is relevant since this gas has high-energy recovery without involving carbon. Before microalgae harvesting, a number of range diameters (1-7 mm) for SAPs spherical particles were tested, and the initial rate (V0 ) and the maximal capacity (Qmax ) were determined for the Sueoka medium absorption. The SAP particles with the diameter range 2.0-2.5 mm performed the best and these were employed for the rest of the experiments. The Sueoka medium has a high salt content and the effect of the ionic strength was also studied for different medium concentrations (0-400%). The SAPs were reused in consecutive absorption/desorption cycles, maintaining their absorption capacity. Although the Sueoka medium reduces the SAPs absorption capacity to 40% compared with deionized water, the use of SAPs was very significant for the desulfurization process of C. reihardtii. The presence of C. reinhardtii at different concentrations does not affect the absorption capacity of the Sueoka culture medium by the SAPs. In order to reduce the time of the process, an increase of the SAPs concentration was tested, being 20 g of SAP per liter of medium, a condition to harvest the microalgae culture in 4 h. There were no evident cell ruptures during the harvesting process and the cells remained alive. Finally, the harvested biomass was re-suspended in TAP-S medium and kept under anaerobic conditions and illumination to produce H2 that was monitored by a PEM fuel cell. The use of SAPs for microalgae harvesting is a feasible non-invasive procedure to obtain high concentrations of functional biomass at low cost; it offers an attractive alternative due to its versatility and simplicity. © 2013 Wiley Periodicals, Inc.

A novel liquid medium for the efficient growth of the salmonid pathogen Piscirickettsia salmonis and optimization of culture conditions.

PubMed

Henríquez, Mirtha; González, Ernesto; Marshall, Sergio H; Henríquez, Vitalia; Gómez, Fernando A; Martínez, Irene; Altamirano, Claudia

2013-01-01

Piscirickettsia salmonis is the bacterium that causes Piscirickettsiosis, a systemic disease of salmonid fish responsible for significant economic losses within the aquaculture industry worldwide. The growth of the bacterium for vaccine formulation has been traditionally accomplished by infecting eukaryotic cell lines, a process that involves high production costs and is time-consuming. Recent research has demonstrated that it is possible to culture pure P. salmonis in a blood containing (cell-free) medium. In the present work we demonstrate the growth of P. salmonis in a liquid medium free from blood and serum components, thus establishing a novel and simplified bacteriological medium. Additionally, the new media reported provides improved growth conditions for P. salmonis, where biomass concentrations of approximately 800 mg cell dry weight L(-1) were obtained, about eight times higher than those reported for the blood containing medium. A 2- level full factorial design was employed to evaluate the significance of the main medium components on cell growth and an optimal temperature range of 23-27°C was determined for the microorganism to grow in the novel liquid media. Therefore, these results represent a breakthrough regarding P. salmonis research in order to optimize pure P. salmonis growth in liquid blood and serum free medium.

[Influence of glucose and galactose on the morphology and biological properties of Yersinia pseudotuberculosis].

PubMed

Bakholdina, S I; Solov'eva, T F; Shubin, F N; Timchenko, N F

2005-01-01

When cultivated in the presence of glucose, irrespective of temperature and the degree of aeration, Y. pseudotuberculosis cells have the ovoid form, constant size and low hydrophobic properties of their surface. Meanwhile the characteristics of the bacteria grown in the medium, carbohydrate-free or with galactose added, essentially depend on the conditions of medium aeration. Under the conditions of intensive stirring at both temperatures these bacteria acquire the coccoid form, not typical for Yersinia, they have a smaller area (approximately 2 times) and more hydrophobic surface in comparison with the cells grown in the presence of glucose. Under stationary conditions the differences between the cells, cultivated in the presence of galactose and glucose, in form and area disappear, but the differences in the hydrophobic properties of the surface are retained. As revealed in this study, the cells grown in the presence of galactose and under the conditions of intensive medium stirring, in contrast to those grown with glucose, have 1.5-fold greater invasive activity, irrespective of aeration conditions, eightfold greater resistance to ampicillin and twofold greater resistance to streptomycin and erythromycin.

Presynaptic elements involved in the maintenance of the neuromuscular junction

NASA Technical Reports Server (NTRS)

Burrows, G. H.

1984-01-01

Alterations in the neuromuscular junction were observed in rats preceding loss of muscle mass. In view of the possibility that these alterations involve changes in the secretion of myotrophic agents by presynaptic motor neurons, an investigation was undertaken to characterize a neuronall factor which is thought to be involved in the initiation and maintenance of cholinergic synapses. This factor, which is secreted into the incubation medium by NG108-15 neuroblastoma x glioma hybrid cells, induces the aggregation of nicotinic acetylcholine receptors on primary cultures of rat hindlimb myotubes. Previous attempts to purify this factor failed. Extensive washing of the NG108-15 cells with hepes-buffered salt solution followed by short (4 hour) collection times resulted in the collection of incubation medium containing maximal aggregation activity with as little as 5 ug secreted protein per ml of fresh medium. A three-fold increase in specific activity was obtained after anion exchange chromatography.

An efficient and reproducible method for in vitro clonal multiplication of Rauvolfia tetraphylla L. and evaluation of genetic stability using DNA-based markers.

PubMed

Faisal, Mohammad; Alatar, Abdulrahman A; Ahmad, Naseem; Anis, Mohammad; Hegazy, Ahmad K

2012-12-01

An efficient protocol is described for the rapid in vitro clonal propagation of an endangered medicinal plant, Rauvolfia tetraphylla L., through high frequency shoot induction from nodal explants collected from young shoots of a field grown plant. Effects of growth regulators [6-benzyladenine (BA), kinetin (Kin) 2iP, or α-naphthalene acetic acid (NAA)], carbohydrates, different medium [Murashige and Skoog (MS), Woody Plant Medium (WPM), Gamborg medium (B5), Linsmier and Skoog medium (LS)], and various pH levels on in vitro morphogenesis were investigated. The highest frequency of shoot regeneration (90 %) and maximum number of shoot (35.4 ± 2.3) per explant were observed on WPM medium supplemented with 7.5 μM BA, 2.5 μM NAA, and 30 g/l sucrose at pH 5.8. Well-developed shoots, 4-5 cm in length, were successfully rooted ex vitro at 90 % by a 30-min pulse treatment with 150 μM IBA prior to their transfer in planting substrates. The survival rate of transplantation reached 90 % when transferred to field condition. Genetic stability of micropropagated plantlets was assessed and compared with mother plant using Random Amplified Polymorphic DNA and Inter Simple Sequence Repeats markers. No variation was observed in DNA fingerprinting patterns among the micropropagated plants, which were similar to that of the donor plant illustrating their genetic uniformity and clonal fidelity. This confirms that clonal propagation of this plant using axillary shoot buds can be used for commercial exploitation of the selected genotype where a high degree of fidelity is an essential prerequisite. The work contributed to a better in vitro regeneration and clonal mass multiplication of R. tetraphylla and to develop a strategy for the germplasm conservation of this endangered medicinal plant.

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.

Sum-frequency nonlinear Cherenkov radiation generated on the boundary of bulk medium crystal.

PubMed

Wang, Xiaojing; Cao, Jianjun; Zhao, Xiaohui; Zheng, Yuanlin; Ren, Huaijin; Deng, Xuewei; Chen, Xianfeng

2015-12-14

We demonstrated experimentally a method to generate the sum-frequency Nonlinear Cherenkov radiation (NCR) on the boundary of bulk medium by using two synchronized laser beam with wavelength of 1300 nm and 800 nm. It is also an evidence that the polarization wave is always confined to the boundary. Critical conditions of surface sum-frequency NCR under normal and anomalous dispersion condition is discussed.

Studies on Batch Production of Bacterial Concentrates from Mixed Species Lactic Starters

PubMed Central

Pettersson, H. E.

1975-01-01

Optimum growth conditions for mixed species starter FDs 0172 at constant pH in skim milk, whey, and tryptone medium were investigated. Growth rate and maximum population were optimal at 30 C. pH values between 5.5 and 7.0 did not influence the growth rate and maximum population obtainable. Lactic acid-producing activity declined rapidly after reaching the end of the exponential growth phase. The bacterial balance was found to be influenced by the growth parameters: media, pH, temperature, and neutralizer. Skim milk or whey medium at 25 C, pH 6.5, and neutralized with 20% (vol/vol) NH4OH kept the bacterial balance almost constant throughout the cultivation. Grown in tryptone medium at constant pH, the changes in bacterial balance and other metabolic activities were striking compared to the other two media tested. The effect of lactate as an inhibitor was found to be complex, changing with the growth conditions. Concentrates made from mixed species starters FDs 0172, FD 0570, CH 0170, CHs 0170, and T 27 were comparable to controls when cultivated at the optimum conditions found and thereafter centrifuged. Aroma production, proteolytic activity, and CO2 production did not change significantly compared to controls when cultivated at optimum conditions in skim milk or whey medium. PMID:16350009

Compatible intracellular ion composition of the host improves carbon assimilation by zooxanthellae in mutualistic symbioses.

PubMed

Seibt, C; Schlichter, D

2001-09-01

Cytosymbiotic algae within the host's plasma are exposed to completely different ionic conditions than microalgae living in the sea. The altered ionic gradients, in particular, could be the reason for higher in hospite carbon assimilation levels. To study the effect of varying extracellular ionic conditions on isolated zooxanthellae, their photosynthetic capacity in pure seawater was compared to that in a test medium in which the concentrations of the major inorganic ions, the pH and the osmolality were adjusted to the conditions measured in the host cytoplasm. In this test medium the ratio between oxygen evolution and carbon fixation was 1.2:1.0; in contrast, zooxanthellae in the hyperionic seawater medium showed a comparatively higher oxygen production (2.6:1.0). These results are attributed to a higher energy demand for ion regulation of the isolated algae in the hyperionic medium. Isolated cytosymbionts in seawater need more energy both for the readjustment to the original intracellular ion concentration within the host cell and also for the maintenance of a much steeper gradient during incubation under hyperionic conditions outside the host. The particular intracellular ion concentration of the host cells could have been a decisive evolutionary factor for the very successful establishment of the mutualistic symbioses between anthozoans and dinoflagellates more than 200 million years ago.

76 FR 29242 - Agency Information Collection Activities: Final Collection; Comment Request

Federal Register 2010, 2011, 2012, 2013, 2014

2011-05-20

... exported goods covered under its short and medium-term export credit insurance policies. It also alerts Ex... insurance policies. It also alerts Ex-Im Bank staff of defaults, so they can manage the portfolio in an...

76 FR 14009 - Agency Information Collection Activities: Final Collection; Comment Request

Federal Register 2010, 2011, 2012, 2013, 2014

2011-03-15

... exported goods covered under its short and medium-term export credit insurance policies. It also alerts Ex... policies. It also alerts Ex-Im Bank staff of defaults, so they can manage the portfolio in an informed...

Conditioning to ethanol in the fruit fly-a study using an inhibitor of ADH.

PubMed

Cadieu, N; Cadieu, J -C.; El Ghadraoui, L; Grimal, A; Lamboeuf, Y

1999-06-01

To identify processes involved in the choice of ethanol by adult Drosophila, flies homozygous Adh(F), reared in the absence of alcohol were placed in contact with: a) an ethanol-free medium, b) a medium containing ethanol, c) a medium supplemented with 4-methylpyrazole (4-MP, an inhibitor of the ADH pathway), d) a medium containing ethanol and 4-MP. The choice of ethanol over a medium without ethanol was evaluated by measuring the duration of extension of the proboscis of the flies in each of the media. A slight preference for the ethanol-supplemented medium was observed in the naive flies, which was enhanced by previous exposure to ethanol. Exposure to ethanol and 4-MP, however, led to an avoidance of ethanol. There was a reduction in ADH activity on treatment of the flies with 4-MP, and signs of malaise (reduced locomotor activity, loss of balance) were observed in the flies who ingested both ethanol and inhibitor. We concluded that the preference for ethanol stems from an associative learning related to ethanol utilization. Inhibition of enzymes of ADH pathway led to a conditioned aversion due to disturbance of ethanol metabolism giving rise to malaise.

Humanities and Social Sciences Books of Ten National Disciplinary Associations, 2000-2009

ERIC Educational Resources Information Center

Wiberley, Stephen E., Jr.

2016-01-01

Books are the most important medium of communication in the humanities, a major medium in the social sciences, and a central component of academic library collections. This study examined humanities and social sciences books that won prizes from ten leading United States disciplinary associations between 2000 and 2009. The study extends earlier…

Amending storage vessel and media improves transfer interval of Musa spp. tissue culture plantlets

USDA-ARS?s Scientific Manuscript database

Musa spp. are some of the most important fruit food crops in the world. The USDA-ARS TARS maintains a Musa spp. germplasm collection of ~150 accessions in field plots and in medium-term storage in vitro. Accessions maintained in vitro require routine sub-culturing as nutrient medium is lost due to ...

Film as a "Thoughtful" Medium for Teaching History

ERIC Educational Resources Information Center

Stoddard, Jeremy D.

2012-01-01

This collective case study of teachers and students in two ninth-grade US history classes examines the role that films can play as a "thoughtful" medium for teaching history. Specifically, the study focuses on the nature and range of authentic intellectual work that students are engaged in with film in the classroom (Newmann, F., B.…

What Happens to Students' English after One Year of English­-Medium Course Study at University?

ERIC Educational Resources Information Center

Gan, Zhengdong; Stapleton, Paul; Yang, Chi Cheung Ruby

2015-01-01

A much under-researched issue in higher education is the extent to which English-medium university courses help students improve their English proficiency in an ESL context. Adopting a longitudinal, mixed methods design in which quantitative and qualitative data were collected and analyzed, the current study tracked English language improvement,…

The Forgotten Voices in Higher Education: Students' Satisfaction with English-Medium Instruction

ERIC Educational Resources Information Center

Karakas, Ali

2017-01-01

This study explores how satisfied Turkish students are with English-medium instruction (EMI) in the context of higher education in an era when EMI universities operate as international brands capitalizing on English as a commodity to vie for more customers, that is, national and international students. Data were collected through a questionnaire…

Real-Time Dynamic Brake Assessment Proof of Concept Final Report

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

Lascurain, Mary Beth; Franzese, Oscar; Capps, Gary J

This proof-of-concept research was performed to explore the feasibility of using real-world braking data from commercial motor vehicles to make a diagnosis of brake condition similar to that of the performance-based brake tester (PBBT). This was done by determining the relationship between pressure and brake force (P-BF), compensating for the gross vehicle weight (GVW). The nature of this P-BF relationship (e.g., low braking force for a given brake application pressure) may indicate brake system problems. In order to determine the relationship between brake force and brake application pressure, a few key parameters of duty cycle information were collected. Because brakingmore » events are often brief, spanning only a few seconds, a sample rate of 10 Hz was needed. The algorithm under development required brake application pressure and speed (from which deceleration was calculated). Accurate weight estimation was also needed to properly derive the braking force from the deceleration. In order to ensure that braking force was the predominant factor in deceleration for the segments of data used in analysis, the data was screened for grade as well. Also, the analysis needed to be based on pressures above the crack pressure. The crack pressure is the pressure below which the individual brakes are not applied due the nature of the mechanical system. This value, which may vary somewhat from one wheel end to another, is approximately 10 psi. Therefore, only pressures 15 psi and above were used in the analysis. The Department of Energy s Medium Truck Duty Cycle research has indicated that under the real-world circumstances of the test vehicle brake pressures of up to approximately 30 psi can be expected. Several different types of data were collected during the testing task of this project. Constant-pressure stopping tests were conducted at several combinations of brake application pressure (15, 20, 25, and 30 psi), load conditions (moderately and fully laden), and speeds (20 and 30 mph). Data was collected at 10 Hz. Standard and stepped-pressure performance-based brake tests with brake pressure transducers were performed for each loading condition. The stepped-pressure test included the constant-pressure intervals of brake application at 15, 20, 25, and 30 psi. The PBBT data files included 10 Hz streaming data collected during the testing of each axle. Two weeks of real-world duty cycle (driving and braking) data was also collected at 10 Hz. Initial analysis of the data revealed that the data collected in the field (i.e., day-to-day operations) provided the same information as that obtained from the controlled tests. Analysis of the data collected revealed a strong linear relationship between brake application pressure and deceleration for given GVWs. As anticipated, initial speed was not found to be a significant factor in the deceleration-pressure relationship, unlike GVW. The positive results obtained from this proof of concept test point to the need for further research to expand this concept. A second phase should include testing over a wider range of speeds and include medium brake application pressures in addition to the low pressures tested in this research. Testing on multiple vehicles would also be of value. This future phase should involve testing to determine how degradation of braking performance affects the pressure-deceleration relationship.« less

Alveolar type II cell-fibroblast interactions, synthesis and secretion of surfactant and type I collagen.

PubMed

Griffin, M; Bhandari, R; Hamilton, G; Chan, Y C; Powell, J T

1993-06-01

During alveolar development and alveolar repair close contacts are established between fibroblasts and lung epithelial cells through gaps in the basement membrane. Using co-culture systems we have investigated whether these close contacts influence synthesis and secretion of the principal surfactant apoprotein (SP-A) by cultured rat lung alveolar type II cells and the synthesis and secretion of type I collagen by fibroblasts. The alveolar type II cells remained cuboidal and grew in colonies on fibroblast feeder layers and on Matrigel-coated cell culture inserts but were progressively more flattened on fixed fibroblast monolayers and plastic. Alveolar type II cells cultured on plastic released almost all their SP-A into the medium by 4 days. Alveolar type II cells cultured on viable fibroblasts or Matrigel-coated inserts above fibroblasts accumulated SP-A in the medium at a constant rate for the first 4 days, and probably recycle SP-A by endocytosis. The amount of mRNA for SP-A was very low after 4 days of culture of alveolar type II cells on plastic, Matrigel-coated inserts or fixed fibroblast monolayers: relatively, the amount of mRNA for SP-A was increased 4-fold after culture of alveolar type II cells on viable fibroblasts. Co-culture of alveolar type II cells with confluent human dermal fibroblasts stimulated by 2- to 3-fold the secretion of collagen type I into the culture medium, even after the fibroblasts' growth had been arrested with mitomycin C. Collagen secretion, by fibroblasts, also was stimulated 2-fold by conditioned medium from alveolar type II cells cultured on Matrigel. The amount of mRNA for type I collagen increased only modestly when fibroblasts were cultured in this conditioned medium. This stimulation of type I collagen secretion diminished as the conditioned medium was diluted out, but at high dilutions further stimulation occurred, indicating that a factor that inhibited collagen secretion also was being diluted out. The conditioned medium contained low levels of IGF-1 and the stimulation of type I collagen secretion was abolished when the conditioned medium was pre-incubated with antibodies to insulin-like growth factor 1 (IGF-1). There are important reciprocal interactions between alveolar type II cells and fibroblasts in co-culture. Direct contacts between alveolar type II cells and fibroblasts appear to have a trophic effect on cultured alveolar type II cells, increasing the levels of mRNA for SP-A. Rat lung alveolar type II cells appear to release a factor (possibly IGF-1) that stimulates type I collagen secretion by fibroblasts.

Collective photonic-plasmonic resonances in noble metal - dielectric nanoparticle hybrid arrays

DOE PAGES

Hong, Yan; Reinhard, Björn M.

2014-10-27

Coherent scattering of gold and silver nanoparticles (NPs) in regular arrays can generate Surface Lattice Resonances (SLRs) with characteristically sharp spectral features. Herein, we investigate collective resonances in compositionally more complex arrays comprising NP clusters and NPs with different chemical compositions at pre-defined lattice sites. We first characterize the impact of NP clustering by exchanging individual gold NPs in the array through dimers of electromagnetically strongly coupled gold NPs. Then, we analyze hybrid arrays that contain both gold metal NP dimers and high refractive index dielectric NPs as building blocks. We demonstrate that the integration of gold NP clusters andmore » dielectric NPs into one array enhances E-field intensities not only in the vicinity of the NPs but also in the ambient medium of the entire array. In addition, this work shows that the ability to integrate multiple building blocks with different resonance conditions in one array provides new degrees of freedom for engineering optical fields in the array plane with variable amplitude and phase.« less

Evaluation of Forest Health Conditions using Unmanned Aircraft Systems (UAS)

NASA Astrophysics Data System (ADS)

Hatfield, M. C.; Heutte, T. M.

2016-12-01

US Forest Service Alaska Region Forest Health Protection (FHP) and University of Alaska Fairbanks, Alaska Center for Unmanned Aircraft Systems Integration (ACUASI) are evaluating capability of Unmanned Aerial Systems (UAS) to monitor forest health conditions in Alaska's Interior Region. In July 2016, the team deployed UAS at locations in the Tanana Valley near Fairbanks in order to familiarize FHP staff with capabilities of UAS for evaluating insect and disease damage. While many potential uses of UAS to evaluate and monitor forest health can be envisioned, this project focused on use of a small UAS for rapid assessment of insect and disease damage. Traditional ground-based methods are limited by distance from ground to canopy and inaccessibility of forest stands due to terrain conditions. Observation from fixed-wing aircraft provide a broad overview of conditions but are limited by minimum safe flying altitude (500' AGL) and aircraft speed ( 100 mph). UAS may provide a crucial bridge to fill in gaps between ground and airborne methods, and offer significant cost savings and greater flexibility over helicopter-based observations. Previous uses of UAS for forest health monitoring are limited - this project focuses on optimizing choice of vehicle, sensors, resolution and area scanned from different altitudes, and use of visual spectrum vs NIR image collection. The vehicle selected was the ACUASI Ptarmigan, a small hexacopter (based on DJI S800 airframe and 3DR autopilot) capable of carrying a 1.5 kg payload for 15 min for close-range environmental monitoring missions. Sites were chosen for conditions favorable to UAS operation and presence of forest insect and disease agents including spruce broom rust, aspen leaf miner, birch leaf roller, and willow leafblotch miner. A total of 29 flights were conducted with 9000+ images collected. Mission variables included camera height, UAS speed, and medium- (Sony NEX-7) vs low-resolution (GoPro Hero) cameras. Invaluable knowledge was gained as to limitations and opportunities for field deployment of UAS relative to local conditions of terrain and forest type. Analysis will include image suitability for orthocorrection and production of stand level image mosaic, with further optimization of image collection parameters to detect known insect- and disease-caused disturbance.

Bioleaching of nickel from spent petroleum catalyst using Acidithiobacillus thiooxidans DSM- 11478.

PubMed

Sharma, Mohita; Bisht, Varsha; Singh, Bina; Jain, Pratiksha; Mandal, Ajoy K; Lal, Banwari; Sarma, Priyangshu M

2015-06-01

The present work deals with optimization of culture conditions and process parameters for bioleaching of spent petroleum catalyst collected from a petroleum refinery. The efficacy of Ni bioleaching from spent petroleum catalyst was determined using pure culture of Acidithiobacillus thiooxidans DSM- 11478. The culture conditions of pH, temperature and headspace volume to media volume ratio were optimized. EDX analysis was done to confirm the presence of Ni in the spent catalyst after roasting it to decoke its surface. The optimum temperature for A. thiooxidans DSM-11478 growth was found to be 32 degrees C. The enhanced recovery of nickel at very low pH was attributed to the higher acidic strength of sulfuric acid produced in the culture medium by the bacterium. During the bioleaching process, 89% of the Ni present in the catalyst waste could be successfully recovered in optimized conditions. This environment friendly bioleaching process proved efficient than the chemical method. Taking leads from the lab scale results, bioleaching in larger volumes (1, 5 and 10 L) was also performed to provide guidelines for taking up this technology for in situ industrial waste management.

Wetting-mediated collective tubulation and pearling in confined vesicular drops of DDAB solutions.

PubMed

Haidara, Hamidou

2014-12-21

Whether driven by external mechanical stresses (shear flow) or induced by membrane-active peptides and/or proteins, the collective growth of tubules in membranous fluids has seldom been reported. The pearling destabilization of these membranous tubules which requires an activation of the shape distortion, often induced by optical tweezers, membrane-active biomolecules or an electrical field, has also rarely been observed under mild experimental conditions. Here we report such events of collective tubulation and pearling destabilization in sessile drops of a didodecyl-dimethylammonium bromide (DDAB) vesicular solution that are confined by a surrounding oil medium. Based on the wetting dynamics and the features of the tubulation process, we show that the growth of the tubules here relies on a mechanism of "pinning-induced pulling" from the retracting drop, rather than the classical hydrodynamic fingering instability. We show that the whole tubulation process is driven by a strong coupling between the bulk properties of the ternary (DAAB/water/oil) system and the dynamics of wetting. Finally, we discuss the pearling destabilization of these tubules under vanishing static interface tension and quite mild tensile force arising from their pulling. We show that under those mild conditions, shape disturbances readily grow, either as pearling waves moving toward the drop-reservoir or as Rayleigh-type peristaltic modulations. Besides revealing singular non-Rayleigh pearling modes, this work also brings new insights into the flow dynamics in membranous tubules anchored to an infinite reservoir.

Effects of mineral trioxide aggregate, BiodentineTM and calcium hydroxide on viability, proliferation, migration and differentiation of stem cells from human exfoliated deciduous teeth

PubMed Central

Araújo, Leandro Borges; Cosme-Silva, Leopoldo; Fernandes, Ana Paula; de Oliveira, Thais Marchini; Cavalcanti, Bruno das Neves; Gomes, João Eduardo; Sakai, Vivien Thiemy

2018-01-01

Abstract Objective The aim of the study was to evaluate the effects of the capping materials mineral trioxide aggregate (MTA), calcium hydroxide (CH) and BiodentineTM (BD) on stem cells from human exfoliated deciduous teeth (SHED) in vitro. Material and Methods SHED were cultured for 1 – 7 days in medium conditioned by incubation with MTA, BD or CH (1 mg/mL), and tested for viability (MTT assay) and proliferation (SRB assay). Also, the migration of serum-starved SHED towards conditioned media was assayed in companion plates, with 8 μm-pore-sized membranes, for 24 h. Gene expression of dentin matrix protein-1 (DMP-1) was evaluated by reverse-transcription polymerase chain reaction. Regular culture medium with 10% FBS (without conditioning) and culture medium supplemented with 20% FBS were used as controls. Results MTA, CH and BD conditioned media maintained cell viability and allowed continuous SHED proliferation, with CH conditioned medium causing the highest positive effect on proliferation at the end of the treatment period (compared with BD and MTA) (p<0.05). In contrast, we observed increased SHED migration towards BD and MTA conditioned media (compared with CH) (p<0.05). A greater amount of DMP-1 gene was expressed in MTA group compared with the other groups from day 7 up to day 21. Conclusion Our results show that the three capping materials are biocompatible, maintain viability and stimulate proliferation, migration and differentiation in a key dental stem cell population. PMID:29412365

75 FR 1620 - Agency Information Collection Activities: Final Collection; Comment Request

Federal Register 2010, 2011, 2012, 2013, 2014

2010-01-12

... the survey: 1. Added question--Years in Business in Part 1, Question 1. 2. Removed ``Medium-term Loan... mandate (Export-Import Act of 1945, as amended, 12 U.S.C. 635) which directs the Export-Import Bank to...

Information system of quality assessment for liquid and gaseous medium production

NASA Astrophysics Data System (ADS)

Bobrov, V. N.; Us, N. A.; Davidov, I. S.

2018-05-01

A method and a technical solution for controlling the quality of production of liquid and gaseous media is proposed. It is also proposed to monitor harmful factors in production while ensuring safe working conditions. Initially, using the mathematical model of an ideal atmosphere, the projection to the horizontal surface of the observation trajectory is calculated. At the second stage, the horizontal projection of the observation trajectory in real conditions is measured. The quality of the medium is judged by the difference between the projections of observation trajectories. The technical result is presented in the form of a device allowing obtaining information about the quality of the medium under investigation.

Phosphorus and nitrogen recycle following algal bio-crude production via continuous hydrothermal liquefaction

DOE PAGES

Edmundson, S.; Huesemann, M.; Kruk, R.; ...

2017-07-25

Phosphorus and nitrogen are essential components of microalgal growth media. Critical to a wide range of biochemical processes, they commonly limit primary productivity. Recycling elemental phosphorus and fixed nitrogen after fuel conversion via hydrothermal liquefaction (HTL) of algae biomass reduces the need for mined phosphorus and synthetic nitrogen resources. We used scenedesmus obliquus DOE 0152.Z and Chlorella sorokiniana DOE1412 as test organisms in assessing nutrient recycle of phosphorus from filtered solids collected downstream of the HTL reactor and nitrogen collected from the aqueous phase after gravimetric biocrude separation. Maximum specific growth rates were measured in growth media using HTL wastemore » as the sole source of either phosphorus or nitrogen and were compared to an algal growth medium control (BG-11). The maximum specific growth rate of both organisms in the recycled phosphorus medium were nearly identical to rates observed in the control medium. Both organisms showed significantly reduced growth rates in the recycled nitrogen medium. C. sorokiniana DOE1412 adapted after several days of exposure whereas S. obliquus DOE0152.Z exhibited poor adaptability to the recycled nitrogen medium. After adaptation, growth rates observed with C. sorokiniana DOE1412 in the recycled nitrogen medium were 3.02 (± 0.13) day -1, 89% of the control medium (3.40 ± 0.21). We further tested maximum specific growth rates of C. sorokiniana DOE1412 in a medium derived entirely from HTL byproducts, completely replacing all components including nitrogen and phosphorus. In this medium we observed rates of 2.70 ± 0.05 day -1, 79% of the control. By adding trace metals to this recycled medium we improved growth rates significantly to 3.10 ± 0.10, 91% of the control, which indicates a critical element is lost in the conversion process. Recycling elemental resources such as phosphorus and nitrogen from the HTL biofuel conversion process can provide a significant reduction in media cost and improves the prospects for industrial scale, algae-based biofuels.« less

Phosphorus and nitrogen recycle following algal bio-crude production via continuous hydrothermal liquefaction

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

Edmundson, S.; Huesemann, M.; Kruk, R.

Phosphorus and nitrogen are essential components of microalgal growth media. Critical to a wide range of biochemical processes, they commonly limit primary productivity. Recycling elemental phosphorus and fixed nitrogen after fuel conversion via hydrothermal liquefaction (HTL) of algae biomass reduces the need for mined phosphorus and synthetic nitrogen resources. We used scenedesmus obliquus DOE 0152.Z and Chlorella sorokiniana DOE1412 as test organisms in assessing nutrient recycle of phosphorus from filtered solids collected downstream of the HTL reactor and nitrogen collected from the aqueous phase after gravimetric biocrude separation. Maximum specific growth rates were measured in growth media using HTL wastemore » as the sole source of either phosphorus or nitrogen and were compared to an algal growth medium control (BG-11). The maximum specific growth rate of both organisms in the recycled phosphorus medium were nearly identical to rates observed in the control medium. Both organisms showed significantly reduced growth rates in the recycled nitrogen medium. C. sorokiniana DOE1412 adapted after several days of exposure whereas S. obliquus DOE0152.Z exhibited poor adaptability to the recycled nitrogen medium. After adaptation, growth rates observed with C. sorokiniana DOE1412 in the recycled nitrogen medium were 3.02 (± 0.13) day -1, 89% of the control medium (3.40 ± 0.21). We further tested maximum specific growth rates of C. sorokiniana DOE1412 in a medium derived entirely from HTL byproducts, completely replacing all components including nitrogen and phosphorus. In this medium we observed rates of 2.70 ± 0.05 day -1, 79% of the control. By adding trace metals to this recycled medium we improved growth rates significantly to 3.10 ± 0.10, 91% of the control, which indicates a critical element is lost in the conversion process. Recycling elemental resources such as phosphorus and nitrogen from the HTL biofuel conversion process can provide a significant reduction in media cost and improves the prospects for industrial scale, algae-based biofuels.« less

Hair-growth-promoting effect of conditioned medium of high integrin α6 and low CD 71 (α6bri/CD71dim) positive keratinocyte cells.

PubMed

Won, Chong Hyun; Jeong, Yun-Mi; Kang, Sangjin; Koo, Tae-Sung; Park, So-Hyun; Park, Ki-Young; Sung, Young-Kwan; Sung, Jong-Hyuk

2015-02-19

Keratinocyte stem/progenitor cells (KSCs) reside in the bulge region of the hair follicles and may be involved in hair growth. Hair follicle dermal papilla cells (HFDPCs) and outer root sheath (ORS) cells were treated with conditioned medium (CM) of KSCs. Moreover, the effects of KSC-CM on hair growth were examined ex vivo and in vivo. A human growth factor chip array and RT-PCR were employed to identify enriched proteins in KSC-CM as compared with CM from keratinocytes. KSC-CM significantly increased the proliferation of HFDPCs and ORS cells, and increased the S-phase of the cell cycle in HFDPCs. KSC-CM led to the phosphorylation of ATK and ERK1/2 in both cell types. After subcutaneous injection of KSC-CM in C3H/HeN mice, a significant increase in hair growth and increased proliferation of hair matrix keratinocytes ex vivo was observed. We identified six proteins enriched in KSC-CM (amphiregulin, insulin-like growth factor binding protein-2, insulin-like growth factor binding protein-5, granulocyte macrophage-colony stimulating factor, Platelet-derived growth factor-AA, and vascular endothelial growth factor). A growth-factor cocktail that contains these six recombinant growth factors significantly increased the proliferation of HFDPCs and ORS cells and enhanced the hair growth of mouse models. These results collectively indicate that KSC-CM has the potential to increase hair growth via the proliferative capacity of HFDPCs and ORS cells.

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

Huang, Yuan; Luo, Fangjun; Li, Hui

During inflammation in the glomerulus, the proliferation of myofiroblast-like mesangial cells is commonly associated with the pathological process. Macrophages play an important role in regulating the growth of resident mesangial cells in the glomeruli. Alternatively activated macrophage (M2 macrophage) is a subset of macrophages induced by IL-13/IL-4, which is shown to play a repair role in glomerulonephritis. Prompted by studies of development, we performed bone marrow derived macrophage and rat mesangial cell co-culture study. Conditioned medium from IL-4 primed M2 macrophages induced rat mesangial cell apoptosis. The pro-apoptotic effect of M2 macrophages was demonstrated by condensed nuclei stained with Hoechstmore » 33258, increased apoptosis rates by flow cytometry analysis and enhanced caspase-3 activation by western blot. Fas protein was up-regulated in rat mesangial cells, and its neutralizing antibody ZB4 partly inhibited M2 macrophage-induced apoptosis. The up-regulated arginase-1 expression in M2 macrophage also contributed to this apoptotic effect. These results indicated that the process of apoptosis triggered by conditioned medium from M2 macrophages, at least is partly conducted through Fas in rat mesangial cells. Our findings provide compelling evidence that M2 macrophages control the growth of mesangial cells in renal inflammatory conditions. - Highlights: • Conditioned-medium from M2 macrophages induces rat mesangial cell (MsC) apoptosis. • M2 macrophage conditioned medium exerts its pro-apoptotic effects via Fas ligand. • Arginase-1 activity in M2 macrophages plays a role in inducing apoptosis in rat MsC.« less

77 FR 73115 - Proposed Collection; Comment Request for Information Collection Tools

Federal Register 2010, 2011, 2012, 2013, 2014

2012-12-07

... section 472(c) or (e)(2). Revenue Procedure 98-46 modified Revenue Procedure 97-44 by allowing medium-and... on proposed and/or continuing information collections, as required by the Paperwork Reduction Act of 1995, Public Law 104-13 (44 U.S.C. 3506(c)(2)(A)). Currently, the IRS is soliciting comments concerning...

Growth, antioxidant capacity and total carotene of Dunaliella salina DCCBC15 in a low cost enriched natural seawater medium.

PubMed

Tran, Duc; Doan, Nguyen; Louime, Clifford; Giordano, Mario; Portilla, Sixto

2014-01-01

Dunaliella is currently drawing worldwide attention as an alternative source of nutraceuticals. Commercially, β-carotene making up over 10% of Dunaliella biomass is generating the most interest. These compounds, because of their non-toxic properties, have found applications in the food, drug and cosmetic industry. The β-carotene content of Dunaliella cells, however, depends heavily on the growth conditions and especially on the availability of nutrients, salinity, irradiance and temperature in the growth medium. A chemically well defined medium is usually required, which significantly contributes to the cost of pigment production; hence a desire for low cost marine media. The present study aimed at evaluating the suitability of six different media, especially exploiting local potential resources, for the mass production of Dunaliella salina DCCBC15 as functional food and medicine. The efficacy of a new selected low-cost enriched natural seawater medium (MD4), supplemented with industrial N-P-K fertilizer, was investigated with respect to biomass production, chlorophyll, antioxidant capacity, and total carotene by Dunaliella though culture conditions were not optimized yet. This new medium (MD4) appears extremely promising, since it affords a higher production of Dunaliella biomass and pigments compared with the control, a common artificial medium (MD1), while allowing a substantial reduction in the production costs. The medium is also recommended for culturing other marine algae.

Exosomes of human placenta-derived mesenchymal stem cells stimulate angiogenesis.

PubMed

Komaki, Motohiro; Numata, Yuri; Morioka, Chikako; Honda, Izumi; Tooi, Masayuki; Yokoyama, Naoki; Ayame, Hirohito; Iwasaki, Kengo; Taki, Atsuko; Oshima, Noriko; Morita, Ikuo

2017-10-03

The therapeutic potential of mesenchymal stem cells (MSCs) may be attributed partly to humoral factors such as growth factors, cytokines, and chemokines. Human term placental tissue-derived MSCs (PlaMSCs), or conditioned medium left over from cultures of these cells, have been reported to enhance angiogenesis. Recently, the exosome, which can transport a diverse suite of macromolecules, has gained attention as a novel intercellular communication tool. However, the potential role of the exosome in PlaMSC therapeutic action is not well understood. The purpose of this study was to evaluate PlaMSC-derived exosome angiogenesis promotion in vitro and in vivo. MSCs were isolated from human term placental tissue by enzymatic digestion. Conditioned medium was collected after 48-h incubation in serum-free medium (PlaMSC-CM). Angiogenic factors present in PlaMSC-CM were screened by a growth factor array. Exosomes were prepared by ultracentrifugation of PlaMSC-CM, and confirmed by transmission electron microscopy, dynamic light scattering, and western blot analyses. The proangiogenic activity of PlaMSC-derived exosomes (PlaMSC-exo) was assessed using an endothelial tube formation assay, a cell migration assay, and reverse transcription-PCR analysis. The in-vivo angiogenic activity of PlaMSC-exo was evaluated using a murine auricle ischemic injury model. PlaMSC-CM contained both angiogenic and angiostatic factors, which enhanced endothelial tube formation. PlaMSC-exo were incorporated into endothelial cells; these exosomes stimulated both endothelial tube formation and migration, and enhanced angiogenesis-related gene expression. Laser Doppler blood flow analysis showed that PlaMSC-exo infusion also enhanced angiogenesis in an in-vivo murine auricle ischemic injury model. PlaMSC-exo enhanced angiogenesis in vitro and in vivo, suggesting that exosomes play a role in the proangiogenic activity of PlaMSCs. PlaMSC-exo may be a novel therapeutic approach for treating ischemic diseases.

Laying sequence interacts with incubation temperature to influence rate of embryonic development and hatching synchrony in a precocial bird.

PubMed

Hepp, Gary R; Kennamer, Robert A

2018-01-01

Incubation starts during egg laying for many bird species and causes developmental asynchrony within clutches. Faster development of late-laid eggs can help reduce developmental differences and synchronize hatching, which is important for precocial species whose young must leave the nest soon after hatching. In this study, we examined the effect of egg laying sequence on length of the incubation period in Wood Ducks (Aix sponsa). Because incubation temperature strongly influences embryonic development rates, we tested the interactive effects of laying sequence and incubation temperature on the ability of late-laid eggs to accelerate development and synchronize hatching. We also examined the potential cost of faster development on duckling body condition. Fresh eggs were collected and incubated at three biologically relevant temperatures (Low: 34.9°C, Medium: 35.8°C, and High: 37.6°C), and egg laying sequences from 1 to 12 were used. Length of the incubation period declined linearly as laying sequence advanced, but the relationship was strongest at medium temperatures followed by low temperatures and high temperatures. There was little support for including fresh egg mass in models of incubation period. Estimated differences in length of the incubation period between eggs 1 and 12 were 2.7 d, 1.2 d, and 0.7 d at medium, low and high temperatures, respectively. Only at intermediate incubation temperatures did development rates of late-laid eggs increase sufficiently to completely compensate for natural levels of developmental asynchrony that have been reported in Wood Duck clutches at the start of full incubation. Body condition of ducklings was strongly affected by fresh egg mass and incubation temperature but declined only slightly as laying sequence progressed. Our findings show that laying sequence and incubation temperature play important roles in helping to shape embryo development and hatching synchrony in a precocial bird.

Laying sequence interacts with incubation temperature to influence rate of embryonic development and hatching synchrony in a precocial bird

PubMed Central

Kennamer, Robert A.

2018-01-01

Incubation starts during egg laying for many bird species and causes developmental asynchrony within clutches. Faster development of late-laid eggs can help reduce developmental differences and synchronize hatching, which is important for precocial species whose young must leave the nest soon after hatching. In this study, we examined the effect of egg laying sequence on length of the incubation period in Wood Ducks (Aix sponsa). Because incubation temperature strongly influences embryonic development rates, we tested the interactive effects of laying sequence and incubation temperature on the ability of late-laid eggs to accelerate development and synchronize hatching. We also examined the potential cost of faster development on duckling body condition. Fresh eggs were collected and incubated at three biologically relevant temperatures (Low: 34.9°C, Medium: 35.8°C, and High: 37.6°C), and egg laying sequences from 1 to 12 were used. Length of the incubation period declined linearly as laying sequence advanced, but the relationship was strongest at medium temperatures followed by low temperatures and high temperatures. There was little support for including fresh egg mass in models of incubation period. Estimated differences in length of the incubation period between eggs 1 and 12 were 2.7 d, 1.2 d, and 0.7 d at medium, low and high temperatures, respectively. Only at intermediate incubation temperatures did development rates of late-laid eggs increase sufficiently to completely compensate for natural levels of developmental asynchrony that have been reported in Wood Duck clutches at the start of full incubation. Body condition of ducklings was strongly affected by fresh egg mass and incubation temperature but declined only slightly as laying sequence progressed. Our findings show that laying sequence and incubation temperature play important roles in helping to shape embryo development and hatching synchrony in a precocial bird. PMID:29373593

Estrogen receptor (ER)α-regulated lipocalin 2 expression in adipose tissue links obesity with breast cancer progression.

PubMed

Drew, Brian G; Hamidi, Habib; Zhou, Zhenqi; Villanueva, Claudio J; Krum, Susan A; Calkin, Anna C; Parks, Brian W; Ribas, Vicent; Kalajian, Nareg Y; Phun, Jennifer; Daraei, Pedram; Christofk, Heather R; Hewitt, Sylvia C; Korach, Kenneth S; Tontonoz, Peter; Lusis, Aldons J; Slamon, Dennis J; Hurvitz, Sara A; Hevener, Andrea L

2015-02-27

Obesity is associated with increased breast cancer (BrCA) incidence. Considering that inactivation of estrogen receptor (ER)α promotes obesity and metabolic dysfunction in women and female mice, understanding the mechanisms and tissue-specific sites of ERα action to combat metabolic-related disease, including BrCA, is of clinical importance. To study the role of ERα in adipose tissue we generated fat-specific ERα knock-out (FERKO) mice. Herein we show that ERα deletion increased adipocyte size, fat pad weight, and tissue expression and circulating levels of the secreted glycoprotein, lipocalin 2 (Lcn2), an adipokine previously associated with BrCA development. Chromatin immunoprecipitation and luciferase reporter studies showed that ERα binds the Lcn2 promoter to repress its expression. Because adipocytes constitute an important cell type of the breast microenvironment, we examined the impact of adipocyte ERα deletion on cancer cell behavior. Conditioned medium from ERα-null adipocytes and medium containing pure Lcn2 increased proliferation and migration of a subset of BrCA cells in culture. The proliferative and promigratory effects of ERα-deficient adipocyte-conditioned medium on BrCA cells was reversed by Lcn2 deletion. BrCA cell responsiveness to exogenous Lcn2 was heightened in cell types where endogenous Lcn2 expression was minimal, but components of the Lcn2 signaling pathway were enriched, i.e. SLC22A17 and 3-hydroxybutyrate dehydrogenase (BDH2). In breast tumor biopsies from women diagnosed with BrCA we found that BDH2 expression was positively associated with adiposity and circulating Lcn2 levels. Collectively these data suggest that reduction of ERα expression in adipose tissue promotes adiposity and is linked with the progression and severity of BrCA via increased adipocyte-specific Lcn2 production and enhanced tumor cell Lcn2 sensitivity. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

Human umbilical cord tissue-derived mesenchymal stromal cells attenuate remodeling after myocardial infarction by proangiogenic, antiapoptotic, and endogenous cell-activation mechanisms

PubMed Central

2014-01-01

Introduction Among the plethora of cells under investigation to restore a functional myocardium, mesenchymal stromal cells (MSCs) have been granted considerable interest. However, whereas the beneficial effects of bone marrow MSCs (BM-MSCs) in the context of the diseased heart are widely reported, data are still scarce on MSCs from the umbilical cord matrix (UCM-MSCs). Herein we report on the effect of UCM-MSC transplantation to the infarcted murine heart, seconded by the dissection of the molecular mechanisms at play. Methods Human umbilical cord tissue-derived MSCs (UCX®), obtained by using a proprietary technology developed by ECBio, were delivered via intramyocardial injection to C57BL/6 females subjected to permanent ligation of the left descending coronary artery. Moreover, medium produced by cultured UCX® preconditioned under normoxia (CM) or hypoxia (CMH) was collected for subsequent in vitro assays. Results Evaluation of the effects upon intramyocardial transplantation shows that UCX® preserved cardiac function and attenuated cardiac remodeling subsequent to myocardial infarction (MI). UCX® further led to increased capillary density and decreased apoptosis in the injured tissue. In vitro, UCX®-conditioned medium displayed (a) proangiogenic activity by promoting the formation of capillary-like structures by human umbilical vein endothelial cells (HUVECs), and (b) antiapoptotic activity in HL-1 cardiomyocytes subjected to hypoxia. Moreover, in adult murine cardiac Sca-1+ progenitor cells (CPCs), conditioned medium enhanced mitogenic activity while activating a gene program characteristic of cardiomyogenic differentiation. Conclusions UCX® preserve cardiac function after intramyocardial transplantation in a MI murine model. The cardioprotective effects of UCX® were attributed to paracrine mechanisms that appear to enhance angiogenesis, limit the extent of the apoptosis, augment proliferation, and activate a pool of resident CPCs. Overall, these results suggest that UCX® should be considered an alternative cell source when designing new therapeutic approaches to treat MI. PMID:24411922

Transdifferentiation of mouse adipose-derived stromal cells into acinar cells of the submandibular gland using a co-culture system

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

Lee, Jingu; Park, Sangkyu; Roh, Sangho, E-mail: sangho@snu.ac.kr

A loss of salivary gland function often occurs after radiation therapy in head and neck tumors, though secretion of saliva by the salivary glands is essential for the health and maintenance of the oral environment. Transplantation of salivary acinar cells (ACs), in part, may overcome the side effects of therapy. Here we directly differentiated mouse adipose-derived stromal cells (ADSCs) into ACs using a co-culture system. Multipotent ADSCs can be easily collected from stromal vascular fractions of adipose tissues. The isolated ADSCs showed positive expression of markers such as integrin beta-1 (CD29), cell surface glycoprotein (CD44), endoglin (CD105), and Nanog. Themore » cells were able to differentiate into adipocytes, osteoblasts, and neural-like cells after 14 days in culture. ADSCs at passage 2 were co-cultured with mouse ACs in AC culture medium using the double-chamber (co-culture system) to avoid mixing the cell types. The ADSCs in this co-culture system expressed markers of ACs, such as α-amylases and aquaporin5, in both mRNA and protein. ADSCs cultured in AC-conditioned medium also expressed AC markers. Cellular proliferation and senescence analyses demonstrated that cells in the co-culture group showed lower senescence and a higher proliferation rate than the AC-conditioned medium group at Days 14 and 21. The results above imply direct conversion of ADSCs into ACs under the co-culture system; therefore, ADSCs may be a stem cell source for the therapy for salivary gland damage. - Highlights: • ADSCs could transdifferentiate into acinar cells (ACs) using ACs co-culture (CCA). • Transdifferentiated ADSCs expressed ACs markers such as α-amylase and aquaporin5. • High proliferation and low senescence were presented in CCA at Day 14. • Transdifferentiation of ADSCs into ACs using CCA may be an appropriate method for cell-based therapy.« less

Hydrolyzable tannins of tamaricaceous plants. IV: Micropropagation and ellagitannin production in shoot cultures of Tamarix tetrandra.

PubMed

Orabi, Mohamed A A; Taniguchi, Shoko; Terabayashi, Susumu; Hatano, Tsutomu

2011-11-01

Shoot cultures of Tamarix tetrandra on Linsmaier-Skoog (LS) agar medium with 30 g l(-1) sucrose, 2.13 mg l(-1) indoleacetic acid and 2.25 mg l(-1) benzyl adenine produced ellagitannins found in intact plants of the Tamaricaceae. This was demonstrated by the isolation of 14 monomeric-tetrameric ellagitannins from the aq. Me2CO extract of the cultured tissues. This is the first report on the production of ellagitannin tetramers by plant tissue culture. The effects of light and certain medium constituents on tissue growth and ellagitannin production were examined. The contents of representative tannins of different types [i.e., tellimagrandin II (monomer), hirtellin A (linear GOG-type dimer), hirtellin B (hellinoyl-type dimer), hirtellin C (macrocyclic-type dimer), and hirtellin T1 (linear GOG-type trimer)] in the resultant tissues in response to these factors were estimated by HPLC, and the optimal condition for production of these tannins were established. Shoots cultured on LS hormone-free medium promoted root development, and regenerated plants could adapt to ordinary soil and climate. Acclimatized and intact T. tetrandra plants that were collected in November and May, respectively, demonstrated seasonal differences in individual ellagitannin contents. HPLC comparison of individual ellagitannin contents in different plant materials (i.e., leaves, stems, and roots) of intact T. tetrandra plants is also reported. The results are discussed with respect to cellular deposition and biosynthetic relationship of tannins. Copyright © 2011 Elsevier Ltd. All rights reserved.

76 FR 81915 - Proposed Information Collection; Comment Request; National Minority Enterprise Development (MED...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-12-29

... promotes the growth and competitiveness of large, medium, and small minority business enterprises by offering management and technical assistance through a network of regional and local business centers... DEPARTMENT OF COMMERCE Minority Business Development Agency Proposed Information Collection...

Insects for breakfast and whales for dinner: the diet and body condition of dingoes on Fraser Island (K’gari)

PubMed Central

Behrendorff, Linda; Leung, Luke K.-P.; McKinnon, Allan; Hanger, Jon; Belonje, Grant; Tapply, Jenna; Jones, Darryl; Allen, Benjamin L.

2016-01-01

Top-predators play stabilising roles in island food webs, including Fraser Island, Australia. Subsidising generalist predators with human-sourced food could disrupt this balance, but has been proposed to improve the overall health of the island’s dingo (Canis lupus dingo) population, which is allegedly ‘starving’ or in ‘poor condition’. We assess this hypothesis by describing the diet and health of dingoes on Fraser Island from datasets collected between 2001 and 2015. Medium-sized mammals (such as bandicoots) and fish were the most common food items detected in dingo scat records. Stomach contents records revealed additional information on diet, such as the occurrence of human-sourced foods. Trail camera records highlighted dingo utilisation of stranded marine fauna, particularly turtles and whales. Mean adult body weights were higher than the national average, body condition scores and abundant-excessive fat reserves indicated a generally ideal-heavy physical condition, and parasite loads were low and comparable to other dingo populations. These data do not support hypotheses that Fraser Island dingoes have restricted diets or are in poor physical condition. Rather, they indicate that dingoes on Fraser Island are capable of exploiting a diverse array of food sources which contributes to the vast majority of dingoes being of good-excellent physical condition. PMID:27009879

Workplace Education Programs in Small and Medium-Sized Michigan Firms. Staff Working Paper 92-13.

ERIC Educational Resources Information Center

Hollenbeck, Kevin; Anderson, William

A project collected data from small and medium-sized firms (employing fewer than 500) in Michigan concerning workplace education programs. It addressed why firms were or were not offering programs, program characteristics, and program impacts on firms and employees. Case studies of 28 businesses were undertaken from May 1991-July 1992 and a…

New Medium for Isolation of Bacteria From Cement Kiln Dust with a Potential to Apply in Bio-Concrete

NASA Astrophysics Data System (ADS)

Alshalif, A. F.; Irwan, J. M.; Othman, N.; Al-Gheethi, A.

2018-04-01

The present study aimed to introduce a new isolation medium named kiln dust medium (KDM) for recovering of bacteria from cement kiln dust with high pH (>pH 11) without the need for nutrients additives. The cement kiln dust samples were collected from five different areas of Cement Industries of Malaysia Berhad (CIMA). The bacterial isolates were recovered on KDM by direct plating technique. The chemical components for all collected samples were identified using X-ray fluorescence (XRF). The primary identification for the bacterial isolates indicated that these bacteria belongs to Bacillus spp. Based on the morphological characteristics. The growth curve of the bacterial strains was monitored using the optical density (OD) with 650 nm wavelength, which in role confirmed that all isolated bacteria had the ability to grow successfully in the proposed medium. The ability of the bacterial strains to grow at high pH reflects their potential in the bio-concrete applications (aerated and non-aerated concrete). These findings indicated that the cement kiln dust samples from Cement Industries represent the most appropriate source for bacteria used in the bioconcrete.

A Novel Strategy to Increase the Proliferative Potential of Adult Human β-Cells While Maintaining Their Differentiated Phenotype

PubMed Central

Aly, Haytham; Rohatgi, Nidhi; Marshall, Connie A.; Grossenheider, Tiffani C.; Miyoshi, Hiroyuki; Stappenbeck, Thaddeus S.; Matkovich, Scot J.; McDaniel, Michael L.

2013-01-01

Our previous studies demonstrated that Wnt/GSK-3/β-catenin and mTOR signaling are necessary to stimulate proliferative processes in adult human β-cells. Direct inhibition of GSK-3, that engages Wnt signaling downstream of the Wnt receptor, increases β-catenin nuclear translocation and β-cell proliferation but results in lower insulin content. Our current goal was to engage canonical and non-canonical Wnt signaling at the receptor level to significantly increase human β-cell proliferation while maintaining a β-cell phenotype in intact islets. We adopted a system that utilized conditioned medium from L cells that expressed Wnt3a, R-spondin-3 and Noggin (L-WRN conditioned medium). In addition we used a ROCK inhibitor (Y-27632) and SB-431542 (that results in RhoA inhibition) in these cultures. Treatment of intact human islets with L-WRN conditioned medium plus inhibitors significantly increased DNA synthesis ∼6 fold in a rapamycin-sensitive manner. Moreover, this treatment strikingly increased human β-cell proliferation ∼20 fold above glucose alone. Only the combination of L-WRN conditioned medium with RhoA/ROCK inhibitors resulted in substantial proliferation. Transcriptome-wide gene expression profiling demonstrated that L-WRN medium provoked robust changes in several signaling families, including enhanced β-catenin-mediated and β-cell-specific gene expression. This treatment also increased expression of Nr4a2 and Irs2 and resulted in phosphorylation of Akt. Importantly, glucose-stimulated insulin secretion and content were not downregulated by L-WRN medium treatment. Our data demonstrate that engaging Wnt signaling at the receptor level by this method leads to necessary crosstalk between multiple signaling pathways including activation of Akt, mTOR, Wnt/β-catenin, PKA/CREB, and inhibition of RhoA/ROCK that substantially increase human β-cell proliferation while maintaining the β-cell phenotype. PMID:23776620

A novel strategy to increase the proliferative potential of adult human β-cells while maintaining their differentiated phenotype.

PubMed

Aly, Haytham; Rohatgi, Nidhi; Marshall, Connie A; Grossenheider, Tiffani C; Miyoshi, Hiroyuki; Stappenbeck, Thaddeus S; Matkovich, Scot J; McDaniel, Michael L

2013-01-01

Our previous studies demonstrated that Wnt/GSK-3/β-catenin and mTOR signaling are necessary to stimulate proliferative processes in adult human β-cells. Direct inhibition of GSK-3, that engages Wnt signaling downstream of the Wnt receptor, increases β-catenin nuclear translocation and β-cell proliferation but results in lower insulin content. Our current goal was to engage canonical and non-canonical Wnt signaling at the receptor level to significantly increase human β-cell proliferation while maintaining a β-cell phenotype in intact islets. We adopted a system that utilized conditioned medium from L cells that expressed Wnt3a, R-spondin-3 and Noggin (L-WRN conditioned medium). In addition we used a ROCK inhibitor (Y-27632) and SB-431542 (that results in RhoA inhibition) in these cultures. Treatment of intact human islets with L-WRN conditioned medium plus inhibitors significantly increased DNA synthesis ∼6 fold in a rapamycin-sensitive manner. Moreover, this treatment strikingly increased human β-cell proliferation ∼20 fold above glucose alone. Only the combination of L-WRN conditioned medium with RhoA/ROCK inhibitors resulted in substantial proliferation. Transcriptome-wide gene expression profiling demonstrated that L-WRN medium provoked robust changes in several signaling families, including enhanced β-catenin-mediated and β-cell-specific gene expression. This treatment also increased expression of Nr4a2 and Irs2 and resulted in phosphorylation of Akt. Importantly, glucose-stimulated insulin secretion and content were not downregulated by L-WRN medium treatment. Our data demonstrate that engaging Wnt signaling at the receptor level by this method leads to necessary crosstalk between multiple signaling pathways including activation of Akt, mTOR, Wnt/β-catenin, PKA/CREB, and inhibition of RhoA/ROCK that substantially increase human β-cell proliferation while maintaining the β-cell phenotype.

The Release of Elements from Dental Casting Alloy into Cell-Culture Medium and Artificial Saliva

PubMed Central

Can, Gülşen; Akpınar, Gül; Aydın, Ahmet

2007-01-01

Objectives The biocompatibility of dental casting alloys is a critical issue because these alloys are in long-term intimate contact with oral tissues. Since the biocompatibility of alloys is not completely known; the release of elements from the alloys has been studied. The aim of this study was to compare the elemental release from dental casting alloy during exposure to artificial saliva and cell-culture medium. Materials and Methods Twenty specimens made from Ni-Cr alloy were provided in the form of 5 mm diameter discs, 2 mm in thickness with a 7 mm stem attached to one face to facilitate handling. Ten of twenty samples were polished separately using a conventional technique. The remaining ten samples were left sandblasted with 50 μm Al203. Ten samples (5 polished, 5 sandblasted) were separately placed into cell-culture wells with Dulbecco’s Modified Eagle’s Medium. The other ten samples were placed separately into cell-culture wells with artificial saliva. The samples were subjected in contact with these medium for 30 days. These medium were collected every 7 days. The cell-culture medium and artificial saliva without alloy samples were subjected to elemental analyses as a control. At the end of the exposure time, Atomic Absorption Spectrometry (AAS) was used to determine the release of elements from the alloys into all collected medium. Statistical analyses were assessed with two-way ANOVA. Results In general, the elemental release occurred with in all medium. The elemental releases of sandblasted alloys were higher than polished alloys. Artificial saliva was found to cause more release from the samples. In both media, Ni released from polished and sandblasted alloys were higher than Cr and Mo. Conlusions The results suggest that the release of elements from the alloys might have correlated with the environments and the surface of dental alloy. PMID:19212482

The release of elements from dental casting alloy into cell-culture medium and artificial saliva.

PubMed

Can, Gülşen; Akpınar, Gül; Aydın, Ahmet

2007-04-01

The biocompatibility of dental casting alloys is a critical issue because these alloys are in long-term intimate contact with oral tissues. Since the biocompatibility of alloys is not completely known; the release of elements from the alloys has been studied. The aim of this study was to compare the elemental release from dental casting alloy during exposure to artificial saliva and cell-culture medium. Twenty specimens made from Ni-Cr alloy were provided in the form of 5 mm diameter discs, 2 mm in thickness with a 7 mm stem attached to one face to facilitate handling. Ten of twenty samples were polished separately using a conventional technique. The remaining ten samples were left sandblasted with 50 mum Al(2)0(3). Ten samples (5 polished, 5 sandblasted) were separately placed into cell-culture wells with Dulbecco's Modified Eagle's Medium. The other ten samples were placed separately into cell-culture wells with artificial saliva. The samples were subjected in contact with these medium for 30 days. These medium were collected every 7 days. The cell-culture medium and artificial saliva without alloy samples were subjected to elemental analyses as a control. At the end of the exposure time, Atomic Absorption Spectrometry (AAS) was used to determine the release of elements from the alloys into all collected medium. Statistical analyses were assessed with two-way ANOVA. In general, the elemental release occurred with in all medium. The elemental releases of sandblasted alloys were higher than polished alloys. Artificial saliva was found to cause more release from the samples. In both media, Ni released from polished and sandblasted alloys were higher than Cr and Mo. The results suggest that the release of elements from the alloys might have correlated with the environments and the surface of dental alloy.

Collection efficiency of a single optical fiber in turbid media.

PubMed

Bargo, Paulo R; Prahl, Scott A; Jacques, Steven L

2003-06-01

If a single optical fiber is used for both delivery and collection of light, two major factors affect the measurement of collected light: (1) the light transport in the medium that describes the amount of light that returns to the fiber and (2) the light coupling to the optical fiber that depends on the angular distribution of photons entering the fiber. We focus on the importance of the latter factor and describe how the efficiency of the coupling depends on the optical properties of the medium. For highly scattering tissues, the efficiency is well predicted by the numerical aperture (NA) of the fiber. For lower scattering, such as in soft tissues, photons arrive at the fiber from deeper depths, and the coupling efficiency could increase twofold to threefold above that predicted by the NA.

Increased efficiency of mammalian somatic cell hybrid production under microgravity conditions during ballistic rocket flight

NASA Technical Reports Server (NTRS)

Schnettler, R.; Gessner, P.; Zimmermann, U.; Neil, G. A.; Urnovitz, H. B.

1989-01-01

The electrofusion of hybridoma cell lines under short-duration microgravity during a flight of the TEXUS 18 Black Brand ballistic sounding rocket at Kiruna, Sweden is reported. The fusion partners, growth medium, cell fusion medium, cell fusion, cell viability in the fusion medium, and postfusion cell culture are described, and the rocket, cell fusion chamber, apparatus, and module are examined. The experimental timeline, the effects of fusion medium and incubation time on cell viability and hybrid yields, and the effect of microgravity on hybrid yields are considered.

Spectral stability of supercontinuum generation in condensed mediums

NASA Astrophysics Data System (ADS)

Wang, Jier; Zhang, Yizhu; Shen, Huifeng; Jiang, Yuhai; Wang, Zhongyang

2017-07-01

The features of the supercontinuum generation (SCG) using intense femtosecond pulses are systematically investigated in condensed mediums [sapphire, BK7 glass, ultraviolet (UV)-fused silica, and fluoride crystals]. By optimizing the experimental conditions and choosing suitable mediums, the bandwidth of the SCG can be extended to the UV regime with good spectral stability. This study demonstrates that materials with high bandgap present high efficiency for SCG, particularly in the short wavelength region. The achievable short wavelength and low power-density threshold of the SCG exhibit complicated correlations with the bandgap of condensed mediums.

EDA-containing fibronectin increases proliferation of embryonic stem cells.

PubMed

Losino, Noelia; Waisman, Ariel; Solari, Claudia; Luzzani, Carlos; Espinosa, Darío Fernández; Sassone, Alina; Muro, Andrés F; Miriuka, Santiago; Sevlever, Gustavo; Barañao, Lino; Guberman, Alejandra

2013-01-01

Embryonic stem cells (ESC) need a set of specific factors to be propagated. They can also grow in conditioned medium (CM) derived from a bovine granulosa cell line BGC (BGC-CM), a medium that not only preserves their main features but also increases ESC´s proliferation rate. The mitogenic properties of this medium were previously reported, ascribing this effect to an alternative spliced generated fibronectin isoform that contains the extra domain A (FN EDA(+)). Here, we investigated if the FN EDA(+) isoform increased proliferation of mouse and human ES cells. We analyzed cell proliferation using conditioned media produced by different mouse embryonic fibroblast (MEF) lines genetically engineered to express FN constitutively including or excluding the EDA domain (FN EDA(-)), and in media supplemented with recombinant peptides containing or not the EDA. We found that the presence of EDA in the medium increased mouse and human ESC's proliferation rate. Here we showed for the first time that this FN isoform enhances ESC's proliferation. These findings suggest a possible conserved behavior for regulation of ES cells proliferation by this FN isoform and could contribute to improve their culturing conditions both for research and cell therapy.

EDA-Containing Fibronectin Increases Proliferation of Embryonic Stem Cells

PubMed Central

Losino, Noelia; Waisman, Ariel; Solari, Claudia; Luzzani, Carlos; Espinosa, Darío Fernández; Sassone, Alina; Muro, Andrés F.; Miriuka, Santiago; Sevlever, Gustavo; Barañao, Lino; Guberman, Alejandra

2013-01-01

Embryonic stem cells (ESC) need a set of specific factors to be propagated. They can also grow in conditioned medium (CM) derived from a bovine granulosa cell line BGC (BGC-CM), a medium that not only preserves their main features but also increases ESC´s proliferation rate. The mitogenic properties of this medium were previously reported, ascribing this effect to an alternative spliced generated fibronectin isoform that contains the extra domain A (FN EDA+). Here, we investigated if the FN EDA+ isoform increased proliferation of mouse and human ES cells. We analyzed cell proliferation using conditioned media produced by different mouse embryonic fibroblast (MEF) lines genetically engineered to express FN constitutively including or excluding the EDA domain (FN EDA-), and in media supplemented with recombinant peptides containing or not the EDA. We found that the presence of EDA in the medium increased mouse and human ESC’s proliferation rate. Here we showed for the first time that this FN isoform enhances ESC’s proliferation. These findings suggest a possible conserved behavior for regulation of ES cells proliferation by this FN isoform and could contribute to improve their culturing conditions both for research and cell therapy. PMID:24244705

Thermally induced degradation of sulfur-containing aliphatic glucosinolates in broccoli sprouts (Brassica oleracea var. italica) and model systems.

PubMed

Hanschen, Franziska S; Platz, Stefanie; Mewis, Inga; Schreiner, Monika; Rohn, Sascha; Kroh, Lothar W

2012-03-07

Processing reduces the glucosinolate (GSL) content of plant food, among other aspects due to thermally induced degradation. Since there is little information about the thermal stability of GSL and formation of corresponding breakdown products, the thermally induced degradation of sulfur-containing aliphatic GSL was studied in broccoli sprouts and with isolated GSL in dry medium at different temperatures as well as in aqueous medium at different pH values. Desulfo-GSL have been analyzed with HPLC-DAD, while breakdown products were estimated using GC-FID. Whereas in the broccoli sprouts structural differences of the GSL with regard to thermal stability exist, the various isolated sulfur-containing aliphatic GSL degraded nearly equally and were in general more stable. In broccoli sprouts, methylsulfanylalkyl GSL were more susceptible to degradation at high temperatures, whereas methylsulfinylalkyl GSL were revealed to be more affected in aqueous medium under alkaline conditions. Besides small amounts of isothiocyanates, the main thermally induced breakdown products of sulfur-containing aliphatic GSL were nitriles. Although they were most rapidly formed at comparatively high temperatures under dry heat conditions, their highest concentrations were found after cooking in acidic medium, conditions being typical for domestic processing.

Cultivation of Spirulina maxima in medium supplemented with sugarcane vinasse.

PubMed

Dos Santos, Raquel Rezende; Araújo, Ofélia de Queiroz Fernandes; de Medeiros, José Luiz; Chaloub, Ricardo Moreira

2016-03-01

The feasibility of sugarcane vinasse as supplement in growth medium of Spirulina maxima was investigated. The cell was cultivated under autotrophic (no vinasse, 70 μmol photons m(-2) s(-1)), heterotrophic (no light, culture medium supplemented with vinasse at 0.1% v/v and 1.0% v/v) and mixotrophic conditions (70 μmol photons m(-2) s(-1), vinasse at 0.1% v/v and 1.0% v/v). These preliminary results suggested a cyclic two-stage cultivation - CTSC, with autotrophic condition during light phase of the photoperiod (12 h, 70-200 μmol photons m(-2) s(-1)) and heterotrophic condition during dark phase (12h, 3.0% v/v vinasse). The adopted CTSC strategy consisted in three cycles with 75% withdrawal of suspension and reposition of medium containing 3.0% v/v vinasse, separated by autotrophic rest periods of few days between cycles. Results show an increase of biomass concentration between 0.495 g L(-1) and 0.609 g L(-1) at the 7th day of each cycle and high protein content (between 74.3% and 77.3% w/w). Copyright © 2016 Elsevier Ltd. All rights reserved.

Born to Yawn? Understanding Yawning as a Warning of the Rise in Cortisol Levels: Randomized Trial

PubMed Central

Bishop, Phil

2012-01-01

Background Yawning consistently poses a conundrum to the medical profession and neuroscientists. Despite neurological evidence such as parakinesia brachialis oscitans in stroke patients and thermo-irregulation in multiple sclerosis patients, there is considerable debate over the reasons for yawning with the mechanisms and hormonal pathways still not fully understood. Cortisol is implicated during yawning and may link many neurological disorders. Evidence was found in support of the Thompson cortisol hypothesis that proposes cortisol levels are elevated during yawning just as they tend to rise during stress and fatigue. Objectives To investigate whether saliva cortisol levels rise during yawning and, therefore, support the Thompson cortisol hypothesis. Methods We exposed 20 male and female volunteers aged between 18 and 53 years to conditions that provoked a yawning response in a randomized controlled trial. Saliva samples were collected at the start and again after the yawning response, or at the end of the stimuli presentations if the participant did not yawn. In addition, we collected electromyographic data of the jaw muscles to determine rest and yawning phases of neural activity. Yawning susceptibility scale, Hospital Anxiety and Depression Scale, General Health Questionnaire, and demographic and health details were also collected from each participant. A comprehensive data set allowed comparison between yawners and nonyawners, as well as between rest and yawning phases. Collecting electromyographic data from the yawning phase is novel, and we hope this will provide new information about neuromuscular activity related to cortisol levels. Exclusion criteria included chronic fatigue, diabetes, fibromyalgia, heart conditions, high blood pressure, hormone replacement therapy, multiple sclerosis, and stroke. We compared data between and within participants. Results In the yawning group, there was a significant difference between saliva cortisol samples (t 10 = –3.071, P = .01). Power and effect size were computed based on repeated-measures t tests for both the yawning and nonyawning groups. There was a medium effect size for the nonyawners group (r = .467) but low power (36%). Results were similar for the yawners group: medium effect size (r = .440) and low power (33%). Conclusions There was significant evidence in support of the Thompson cortisol hypothesis that suggests cortisol levels are elevated during yawning. A further longitudinal study is planned to test neurological patients. We intend to devise a diagnostic tool based on changes in cortisol levels that may assist in the early diagnosis of neurological disorders based on the data collected. Trial Registration International Standard Randomized Controlled Trial Number (ISRCTN): 61942768; http://www.controlled-trials.com/ISRCTN61942768/61942768 (Archived by WebCite at http://www.webcitation.org/6A75ZNYvr) PMID:23611879

Paddy farmer households’ participation and food security level in special effort program in Seputih Raman sub-district of Central Lampung Regency

NASA Astrophysics Data System (ADS)

Rangga, K. K.; Syarief, Y. A.

2018-03-01

The objectives of this study are to study the participation of paddy farmers in the Special Effort program to increase paddy production, to study the level of household food security of paddy farmers, and to analyze the correlation between farmer participation and food security level of paddy farmers. The location was chosen purposively in Seputih Raman sub-district. The data were collected from December 2016 to February 2017. The population of this study was paddy farmers who participating in Special Effort program. The hypothesis was tested by using Spearman’s Rank correlation test. Farmer household’s food security was measured objectively based on the share of household’s food expenditure and subjectively based on the opinion, views, and attitudes or farmers’ opinions on food availability, food distribution, and household food consumption. This research showed that farmers’ participation in Special Effort program in Seputih Raman Sub Ditrict, Central Lampung Regency belonged to medium classification, household food security either objectively or subjectively was in food resistant condition of medium classification, and there was significant correlation between farmers’ participation and food security level of paddy farmer household.

Characterization of Residual Medium Peptides from Yersinia pestis Cultures

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

Clowers, Brian H.; Wunschel, David S.; Kreuzer, Helen W.

2013-04-03

Using a range of common microbial medium formulations (TSB, BHI, LB, and G-media), two attenuated strains of Y. pestis (KIM D27 (pgm-) and KIMD1 lcr-) were cultivated in triplicate. These cellular suspensions were used to develop a method of extracting residual medium peptides from the final microbial preparation to assess their relative abundance and identity. Across the conditions examined, which included additional cellular washing and different forms of microbial inactivation, residual medium peptides were detected. Despite the range of growth medium sources used and the associated manufacturing processes used in their production, a high degree of peptide similarity was observedmore » for a given medium recipe. These results demonstrate that residual medium peptides are retained using traditional microbial cultivation techniques and may be used to inform forensic investigations with respect to production deduction.« less

Pretreatments, conditioned medium and co-culture increase the incidence of somatic embryogenesis of different Cichorium species

PubMed Central

Couillerot, Jean-Paul; Windels, David; Vazquez, Franck; Michalski, Jean-Claude; Hilbert, Jean-Louis; Blervacq, Anne-Sophie

2012-01-01

Somatic embryogenesis (SE) in Cichorium involves dedifferentiation and redifferentiation of single cells and can be induced by specific in vitro culture conditions. We have tested the effect of various treatments on the incidence of SE (ISE) of an interspecific embryogenic hybrid (C. endivia x C. intybus) and of different commercial chicories (C. endivia and C. intybus) that are typically recalcitrant to SE in standard culture conditions. We found that the ISE of the hybrid is significantly increased by pretreatment of tissues by submersion in solutions of glycerol, abscisic acid, spermine, putrescine or of combinations of these compounds. Interestingly, the most efficient of these pretreatments also had an unexpectedly high effect on the ISE of the C. intybus cultivars. The ISE of the hybrid and of the commercial chicories were increased when explants were co-cultured with highly embryogenic chicory explants or when they were cultured in conditioned medium. These observations established that unidentified SE-promoting factors are released in the culture medium. HPLC analyses of secreted Arabino-Galactan Proteins (AGPs), which are known to stimulate SE, did not allow identifying a fraction containing differentially abundant AGP candidates. However, pointing to their role in promoting SE, we found that the hybrid had a drastically higher ISE when amino sugars and L-Proline, the putative precursors of secreted AGPs, were both added to the medium. PMID:22301978

Microenvironmental influence on microtumour infiltration patterns: 3D-mathematical modelling supported by in vitro studies.

PubMed

Luján, Emmanuel; Soto, Daniela; Rosito, María S; Soba, Alejandro; Guerra, Liliana N; Calvo, Juan C; Marshall, Guillermo; Suárez, Cecilia

2018-05-09

Mathematical modelling approaches have become increasingly abundant in cancer research. Tumour infiltration extent and its spatial organization depend both on the tumour type and stage and on the bio-physicochemical characteristics of the microenvironment. This sets a complex scenario that often requires a multidisciplinary and individually adjusted approach. The ultimate goal of this work is to present an experimental/numerical combined method for the development of a three-dimensional mathematical model with the ability to reproduce the growth and infiltration patterns of a given avascular microtumour in response to different microenvironmental conditions. The model is based on a diffusion-convection reaction equation that considers logistic proliferation, volumetric growth, a rim of proliferative cells at the tumour surface, and invasion with diffusive and convective components. The parameter values of the model were fitted to experimental results while radial velocity and diffusion coefficients were made spatially variable in a case-specific way through the introduction of a shape function and a diffusion-limited-aggregation (DLA)-derived fractal matrix, respectively, according to the infiltration pattern observed. The in vitro model consists of multicellular tumour spheroids (MTSs) of an epithelial mammary tumour cell line (LM3) immersed in a collagen I gel matrix with a standard culture medium ("naive" matrix) or a conditioned medium from adipocytes or preadipocytes ("conditioned" matrix). It was experimentally determined that both adipocyte and preadipocyte conditioned media had the ability to change the MTS infiltration pattern from collective and laminar to an individual and atomized one. Numerical simulations were able to adequately reproduce qualitatively and quantitatively both kinds of infiltration patterns, which were determined by area quantification, analysis of fractal dimensions and lacunarity, and Bland-Altman analysis. These results suggest that the combined approach presented here could be established as a new framework with interesting potential applications at both the basic and clinical levels in the oncology area.

Conditioned medium from persistently RSV-infected macrophages alters transcriptional profile and inflammatory response of non-infected macrophages.

PubMed

Rivera-Toledo, Evelyn; Salido-Guadarrama, Iván; Rodríguez-Dorantes, Mauricio; Torres-González, Laura; Santiago-Olivares, Carlos; Gómez, Beatriz

2017-02-15

Cells susceptible to persistent viral infections undergo important changes in their biological functions as a consequence of the expression of viral gene products that are capable of altering the gene expression profile of the host cell. Previously, we reported that persistence of the RSV genome in a mouse macrophage cell line induces important alterations in cell homeostasis, including constitutive expression of IFN-β and other pro-inflammatory cytokines. Here, we postulated that changes in the homeostasis of non-infected macrophages could be induced by soluble factors secreted by persistently RSV- infected macrophages. To test this hypothesis, non-infected mouse macrophages were treated with conditioned medium (CM) collected from cultures of persistently RSV-infected macrophages. Total RNA was extracted and a microarray-based gene expression analysis was performed. Non-infected macrophages, treated under similar conditions with CM obtained from cultures of non-infected macrophages, were used as a control to establish differential gene expression between the two conditions. Results showed that CM from the persistently RSV-infected cultures altered expression of a total of 95 genes in non-infected macrophages, resulting in an antiviral gene-transcription profile along with inhibition of the inflammatory response, since some inflammatory genes were down-regulated, including Nlrp3 and Il-1 β, both related to the inflammasome pathway. However, down-regulation of Nlrp3 and Il-1 β was reversible upon acute RSV infection. Additionally, we observed that the inflammatory response, evaluated by secreted IL-1 β, a final product of the inflammasome activity, was enhanced during acute RSV infection in macrophages treated with CM from persistently RSV-infected cultures, compared to that in macrophages treated with the control CM. This suggests that soluble factors secreted during RSV persistence may induce an exacerbated inflammatory response in non-infected cells. Copyright © 2017 Elsevier B.V. All rights reserved.

A complete collection of single-gene deletion mutants of Acinetobacter baylyi ADP1

PubMed Central

de Berardinis, Véronique; Vallenet, David; Castelli, Vanina; Besnard, Marielle; Pinet, Agnès; Cruaud, Corinne; Samair, Sumitta; Lechaplais, Christophe; Gyapay, Gabor; Richez, Céline; Durot, Maxime; Kreimeyer, Annett; Le Fèvre, François; Schächter, Vincent; Pezo, Valérie; Döring, Volker; Scarpelli, Claude; Médigue, Claudine; Cohen, Georges N; Marlière, Philippe; Salanoubat, Marcel; Weissenbach, Jean

2008-01-01

We have constructed a collection of single-gene deletion mutants for all dispensable genes of the soil bacterium Acinetobacter baylyi ADP1. A total of 2594 deletion mutants were obtained, whereas 499 (16%) were not, and are therefore candidate essential genes for life on minimal medium. This essentiality data set is 88% consistent with the Escherichia coli data set inferred from the Keio mutant collection profiled for growth on minimal medium, while 80% of the orthologous genes described as essential in Pseudomonas aeruginosa are also essential in ADP1. Several strategies were undertaken to investigate ADP1 metabolism by (1) searching for discrepancies between our essentiality data and current metabolic knowledge, (2) comparing this essentiality data set to those from other organisms, (3) systematic phenotyping of the mutant collection on a variety of carbon sources (quinate, 2-3 butanediol, glucose, etc.). This collection provides a new resource for the study of gene function by forward and reverse genetic approaches and constitutes a robust experimental data source for systems biology approaches. PMID:18319726

Ionizing radiation-induced bystander mutagenesis and adaptation: Quantitative and temporal aspects

PubMed Central

Zhang, Ying; Zhou, Junqing; Baldwin, Joseph; Held, Kathryn D; Prise, Kevin M; Redmond, Robert W.; Liber, Howard L.

2009-01-01

This work explores several quantitative aspects of radiation-induced bystander mutagenesis in WTK1 human lymphoblast cells. Gamma-irradiation of cells was used to generate conditioned medium containing bystander signals, and that medium was transferred onto naïve recipient cells. Kinetic studies revealed that it required up to one hour to generate sufficient signal to induce the maximal level of mutations at the thymidine kinase locus in the bystander cells receiving the conditioned medium. Furthermore, it required at least one hour of exposure to the signal in the bystander cells to induce mutations. Bystander signal was fairly stable in the medium, requiring 12–24 hours to diminish. Medium that contained bystander signal was rendered ineffective by a 4-fold dilution; in contrast a greater than 20-fold decrease in the cell number irradiated to generate a bystander signal was needed to eliminate bystander-induced mutagenesis. This suggested some sort of feedback inhibition by bystander signal that prevented the signaling cells from releasing more signal. Finally, an ionizing radiation-induced adaptive response was shown to be effective in reducing bystander mutagenesis; in addition, low levels of exposure to bystander signal in the transferred medium induced adaptation that was effective in reducing mutations induced by subsequent γ-ray exposures. PMID:19695271

32 CFR 310.15 - General considerations.

Code of Federal Regulations, 2011 CFR

2011-07-01

... individual's rights, privileges, or benefits under any Federal program. (b) Collecting social security... regardless of the medium used to collect the information (forms, personal interviews, telephonic interviews... mandatory (See paragraph (e) of this section); and (v) The effects on the individual if he or she chooses...

32 CFR 310.15 - General considerations.

Code of Federal Regulations, 2012 CFR

2012-07-01

... individual's rights, privileges, or benefits under any Federal program. (b) Collecting social security... regardless of the medium used to collect the information (forms, personal interviews, telephonic interviews... mandatory (See paragraph (e) of this section); and (v) The effects on the individual if he or she chooses...

32 CFR 310.15 - General considerations.

Code of Federal Regulations, 2013 CFR

2013-07-01

... individual's rights, privileges, or benefits under any Federal program. (b) Collecting social security... regardless of the medium used to collect the information (forms, personal interviews, telephonic interviews... mandatory (See paragraph (e) of this section); and (v) The effects on the individual if he or she chooses...

Ligninolytic enzyme production in selected sub-tropical white rot fungi under different culture conditions.

PubMed

Tekere, M; Zvauya, R; Read, J S

2001-01-01

Lignin peroxidase (LiP), manganese peroxidase (MnP) and laccase activities in selected sub-tropical white rot fungal species from Zimbabwe were determined. The enzyme activities were assayed at varying concentrations of C, N and Mn2+. Manganese peroxidase and laccase activities were the only expressed activities in the fungi under the culture conditions tested. Trametes species, T. cingulata, T. elegans and T. pocas produced the highest manganese peroxidase activities in a medium containing high carbon and low nitrogen conditions. High nitrogen conditions favoured high manganese peroxidase activity in DSPM95, L. velutinus and Irpex spp. High manganese peroxidase activity was notable for T. versicolor when both carbon and nitrogen in the medium were present at high levels. Laccase production by the isolates was highest under conditions of high nitrogen and those conditions with both nitrogen and carbon at high concentration. Mn2+ concentrations between 11-25 ppm gave the highest manganese peroxidase activity compared to a concentration of 40 ppm or when there was no Mn2+ added. Laccase activity was less influenced by Mn2+ levels. While some laccase activity was produced in the absence of Mn2+, the enzyme levels were higher when Mn2+ was added to the culture medium.

Colonization by Legionella spp. of water networks in residential buildings of the Province of Pisa, Italy.

PubMed

Baggiani, A; Casini, B; Totaro, M; Aquino, F; Valentini, P; Bruni, B; Porretta, A; Casalini, F; Miccoli, M; Privitera, G

2015-01-01

Despite the increase of community acquired cases of legionellosis in Italy over the last years, the Italian guidelines do not give indications for prevention and control of Legionella in the hot water networks (or centralized conditioning systems) of residential buildings. We performed a survey on eight medium sized apartment buildings in the Pisa district to assess the prevalence of Legionella spp. in the water network and the respondance to drinking water requisites at the point of use, according to the Italian norms. For each building two hot water and three cold water samples (located at water entrance from the aqueduct network into the building pipework, at the exit from pressure autoclave, and at a remote tap) were collected. Legionella was detected in 20% of residential buildings, mostly in those with a central hot water production system. The study highlights a condition of potential risk for susceptible population subgroups and supports the need for measures of risk assessment and control.

Mechanism of S100b release from rat cortical slices determined under basal and stimulated conditions.

PubMed

Gürsoy, Murat; Büyükuysal, R Levent

2010-03-01

Incubation of rat cortical slices in a medium that was not containing oxygen and glucose (oxygen-glucose deprivation, OGD) caused a 200% increase in the release of S100B. However, when slices were transferred to a medium containing oxygen and glucose (reoxygenation conditions, or REO), S100B release reached 500% of its control value. Neither inhibition of nitric oxide (NO) synthase by L-NAME nor addition of the NO donors sodium nitroprussid (SNP) or hydroxylamine (HA) to the medium altered basal S100B release. Similarly, the presence of SNP, HA or NO precursor L: -arginine in the medium, or inhibition of NO synthase by L-NAME also failed to alter OGD- and REO-induced S100B outputs. Moreover, individual inhibition of PKC, PLA(2) or PLC all failed to attenuate the S100B release determined under control condition or enhanced by either OGD or REO. Blockade of calcium channels with verapamil, chelating the Ca(+2) ions with BAPTA or blockade of sodium channels with tetrodotoxin (TTX) did not alter OGD- and REO-induced S100B release. In contrast to the pharmacologic manipulations mentioned above, glutamate and alpha-ketoglutarate added at high concentrations to the medium prevented both OGD- and REO-induced S100B outputs. These results indicate that neither NO nor the activation of PKC, PLA(2) or PLC seem to be involved in basal or OGD- and REO-induced S100B outputs. Additionally, calcium and sodium currents that are sensitive to verapamil and TTX, respectively, are unlikely to contribute to the enhanced S100B release observed under these conditions.

Chloride-dependency of amyloid beta protein-induced enhancement of glutamate neurotoxicity in cultured rat hippocampal neurons.

PubMed

Zhang, Nan-Yan; Kitagawa, Kaori; Wu, Bo; Xiong, Zheng-Mei; Otani, Hitomi; Inagaki, Chiyoko

2006-05-15

In our previous studies, pathophysiological concentrations of amyloid-beta (Abeta) proteins increased intracellular Cl(-) concentration ([Cl(-)]i) and enhanced glutamate neurotoxicity in primary cultured neurons, suggesting Cl(-)-dependent changes in glutamate signaling. To test this possibility, we examined the effects of isethionate-replaced low Cl(-) medium on the Abeta-induced enhancement of glutamate neurotoxicity in the primary cultured rat hippocampal neurons. In a normal Cl(-) (135 mM) medium, treatment with 10 nM Abeta25-35 for 2 days increased neuronal [Cl(-)]i to a level three times higher than that of control as assayed using a Cl(-)-sensitive fluorescent dye, while in a low Cl(-) (16 mM) medium such an Abeta25-35-induced increase in [Cl(-)]i was not observed. The Abeta treatment aggravated glutamate neurotoxicity in a normal Cl(-) medium as measured by mitochondrial reducing activity and lactate dehydrogenase (LDH) release, while in a low Cl(-) medium the Abeta treatment did not enhance glutamate toxicity. Upon such Abeta plus glutamate treatment under a normal Cl(-) condition, activated anti-apoptotic molecule Akt (Akt-pS473) level monitored by Western blot significantly decreased to 74% of control. Under a low Cl(-) condition, a resting Akt-pS473 level was higher than that under a normal Cl(-) condition and did not significantly change upon Abeta plus glutamate treatment. Tyrosine phosphorylation levels of 110 and 60 kDa proteins (pp110 and pp60) increased upon Abeta plus glutamate treatment under a normal Cl(-), but not low Cl(-), condition. These findings indicated that Abeta-induced enhancement of glutamate neurotoxicity is Cl(-)-dependent. Chloride-sensitive Akt pathway and tyrosine phosphorylation of proteins (pp110 and pp60) may be involved in this process.

Comparison of Four Protocols to Generate Chondrocyte-Like Cells from Human Induced Pluripotent Stem Cells (hiPSCs).

PubMed

Suchorska, Wiktoria Maria; Augustyniak, Ewelina; Richter, Magdalena; Trzeciak, Tomasz

2017-04-01

Stem cells (SCs) are a promising approach to regenerative medicine, with the potential to treat numerous orthopedic disorders, including osteo-degenerative diseases. The development of human-induced pluripotent stem cells (hiPSCs) has increased the potential of SCs for new treatments. However, current methods of differentiating hiPSCs into chondrocyte-like cells are suboptimal and better methods are needed. The aim of the present study was to assess four different chondrogenic differentiation protocols to identify the most efficient method of generating hiPSC-derived chondrocytes. For this study, hiPSCs were obtained from primary human dermal fibroblasts (PHDFs) and differentiated into chondrocyte-like cells using four different protocols: 1) monolayer culture with defined growth factors (GF); 2) embryoid bodies (EBs) in a chondrogenic medium with TGF-β3 cells; 3) EBs in chondrogenic medium conditioned with human chondrocytes (HC-402-05a cell line) and 4) EBs in chondrogenic medium conditioned with human chondrocytes and supplemented with TGF-β3. The cells obtained through these four protocols were evaluated and compared at the mRNA and protein levels. Although chondrogenic differentiation of hiPSCs was successfully achieved with all of these protocols, the two fastest and most cost-effective methods were the monolayer culture with GFs and the medium conditioned with human chondrocytes. Both of these methods are superior to other available techniques. The main advantage of the conditioned medium is that the technique is relatively simple and inexpensive while the directed method (i.e., monolayer culture with GFs) is faster than any protocol described to date because it is does not require additional steps such as EB formation.

Antigenicity of mesenchymal stem cells in an inflamed joint environment.

PubMed

Hill, Jacqueline A; Cassano, Jennifer M; Goodale, Margaret B; Fortier, Lisa A

2017-07-01

OBJECTIVE To determine whether major histocompatability complex (MHC) class II expression in equine mesenchymal stem cells (MSCs) changes with exposure to a proinflammatory environment reflective of an inflamed joint. SAMPLE Cryopreserved bone marrow-derived MSCs from 12 horses and cartilage and synovium samples from 1 horse euthanized for reasons other than lameness. PROCEDURES In part 1 of a 3-part study, the suitability of a quantitative reverse transcriptase PCR (qRT-PCR) assay for measurement of MHC class II expression in MSCs following stimulation with interferon (IFN)-γ was assessed. In part 2, synoviocyte-cartilage cocultures were or were not stimulated with interleukin (IL)-1β (10 ng/mL) to generate conditioned media that did and did not (control) mimic an inflamed joint environment. In part 3, a qRT-PCR assay was used to measure MSC MHC class II expression after 96 hours of incubation with 1 of 6 treatments (control-conditioned medium, IL-1β-conditioned medium, and MSC medium alone [untreated control] or with IL-1β [10 ng/mL], tumor necrosis factor-α [10 ng/mL], or IFN-γ [100 ng/mL]). RESULTS The qRT-PCR assay accurately measured MHC class II expression. Compared with MHC class II expression for MSCs exposed to the untreated control medium, that for MSCs exposed to IL-1β was decreased, whereas that for MSCs exposed to IFN-γ was increased. Neither the control-conditioned nor tumor necrosis factor-α medium altered MHC class II expression. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that MSC exposure to proinflammatory cytokine IL-1β decreased MHC class II expression and antigenicity. Treatment of inflamed joints with allogeneic MSCs might not be contraindicated, but further investigation is warranted.

Competition strategies for the decolorization of a textile-reactive dye with the white-rot fungi Trametes versicolor under non-sterile conditions.

PubMed

Libra, Judy A; Borchert, Maren; Banit, Salima

2003-06-20

A variety of white-rot fungi can oxidize textile dyes under sterile conditions; however, an important consideration for their use in treating wastewater containing textile dyes is whether similar degrees of treatment can be achieved under non-sterile conditions. Four strategies were investigated for their potential in optimizing the use of the fungus Trametes versicolor in non-sterile culture for treating wastewater containing the diazo textile dye C.I. Reactive Black 5 (RB5). Three strategies with suspended culture were designed to increase the decolorization activity in suspended culture from a given amount of T. versicolor inoculum based on its tolerance of low pH (pH reduction in medium), production of extracellular enzymes (use of suspended enzymes alone), and its ability to produce enzymes independent of growth (nitrogen limitation in medium). The results showed that reduction of the medium pH to 3 did not suppress bacterial growth, while enzyme production by T. versicolor ceased. The use of the extracellular enzymes alone would allow the decoupling of the process of fungal growth from wastewater treatment; however, the enzyme activity of an enzyme suspension decreased rapidly under non-sterile conditions. The strategy of limiting nitrogen in the medium to suppress bacterial growth has potential together with the fourth strategy, the cultivation of fungi on organic solids to produce inocula for a decolorization process under non-sterile conditions. A high degree of decolorization of RB5 under non-sterile conditions was achieved with T. versicolor grown on grains as sole substrate. The rate of decolorization was dependent on the amount of fungal inoculum used. Copyright 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 82: 736-744, 2003.

Acanthamoeba and bacteria produce antimicrobials to target their counterpart

PubMed Central

2014-01-01

Background In the microbial ecosystem, microbes compete for space and nutrients. Consequently, some have developed the ability to kill or inhibit the growth of other competing microbes by producing antimicrobial substances. As the ‘producer’ species are generally immune to these substances, their compounds act on the competing microbial species and give the producer more space and access to nutrients for growth. Many currently used antibiotics were developed by exploiting this potential of certain microbes. Findings Here, the free-living amoeba, Acanthamoeba castellanii, was investigated for its antibacterial activity against representative Gram positive and Gram negative bacteria, while bacterial isolates were tested for their anti-amoebic properties. Conditioned medium from A. castellanii showed remarkable bactericidal properties against methicillin-resistant Staphylococcus aureus (MRSA) exhibiting almost 100% kill rate, but had limited effect against Acinetobacter sp., Pseudomonas aeruginosa and vancomycin-resistant Enterococcus faecalis (VRE). Similarly, the conditioned medium of E. coli K1 and Enterobacter sp., exhibited potent anti-Acanthamoebic effects in a concentration-dependent manner. Conditioned media of Acanthamoeba, E. coli K1 and Enterobacter sp. showed no cytotoxicity in vitro when tested against human brain microvascular endothelial cells. Active molecule/s in aforementioned amoebic and two bacterial conditioned media were 5 – 10 kDa, and <5 kDa respectively. Conclusions A. castellanii conditioned medium showed potent bactericidal properties against MRSA. The active molecule(s) are heat- and pronase-resistant, and in the 5 to 10 kDa molecular mass range. Contrary to this, E. coli K1 and Enterobacter sp., conditioned medium showed anti-amoebic effects that are <5 kDa in molecular mass, suggestive of active metabolites. PMID:24479709

Adipose stem cells can secrete angiogenic factors that inhibit hyaline cartilage regeneration.

PubMed

Lee, Christopher Sd; Burnsed, Olivia A; Raghuram, Vineeth; Kalisvaart, Jonathan; Boyan, Barbara D; Schwartz, Zvi

2012-08-24

Adipose stem cells (ASCs) secrete many trophic factors that can stimulate tissue repair, including angiogenic factors, but little is known about how ASCs and their secreted factors influence cartilage regeneration. Therefore, the aim of this study was to determine the effects ASC-secreted factors have in repairing chondral defects. ASCs isolated from male Sprague Dawley rats were cultured in monolayer or alginate microbeads supplemented with growth (GM) or chondrogenic medium (CM). Subsequent co-culture, conditioned media, and in vivo cartilage defect studies were performed. ASC monolayers and microbeads cultured in CM had decreased FGF-2 gene expression and VEGF-A secretion compared to ASCs cultured in GM. Chondrocytes co-cultured with GM-cultured ASCs for 7 days had decreased mRNAs for col2, comp, and runx2. Chondrocytes treated for 12 or 24 hours with conditioned medium from GM-cultured ASCs had reduced sox9, acan, and col2 mRNAs; reduced proliferation and proteoglycan synthesis; and increased apoptosis. ASC-conditioned medium also increased endothelial cell tube lengthening whereas conditioned medium from CM-cultured ASCs had no effect. Treating ASCs with CM reduced or abolished these deleterious effects while adding a neutralizing antibody for VEGF-A eliminated ASC-conditioned medium induced chondrocyte apoptosis and restored proteoglycan synthesis. FGF-2 also mitigated the deleterious effects VEGF-A had on chondrocyte apoptosis and phenotype. When GM-grown ASC pellets were implanted in 1 mm non-critical hyaline cartilage defects in vivo, cartilage regeneration was inhibited as evaluated by radiographic and equilibrium partitioning of an ionic contrast agent via microCT imaging. Histology revealed that defects with GM-cultured ASCs had no tissue ingrowth from the edges of the defect whereas empty defects and defects with CM-grown ASCs had similar amounts of neocartilage formation. ASCs must be treated to reduce the secretion of VEGF-A and other factors that inhibit cartilage regeneration, which can significantly influence how ASCs are used for repairing hyaline cartilage.

Progesterone from the Cumulus Cells Is the Sperm Chemoattractant Secreted by the Rabbit Oocyte Cumulus Complex

PubMed Central

Guidobaldi, Héctor Alejandro; Teves, María Eugenia; Uñates, Diego Rafael; Anastasía, Agustín; Giojalas, Laura Cecilia

2008-01-01

Sperm chemotaxis in mammals have been identified towards several female sources as follicular fluid (FF), oviduct fluid, and conditioned medium from the cumulus oophorus (CU) and the oocyte (O). Though several substances were confirmed as sperm chemoattractant, Progesterone (P) seems to be the best chemoattractant candidate, because: 1) spermatozoa express a cell surface P receptor, 2) capacitated spermatozoa are chemotactically attracted in vitro by gradients of low quantities of P; 3) the CU cells produce and secrete P after ovulation; 4) a gradient of P may be kept stable along the CU; and 5) the most probable site for sperm chemotaxis in vivo could be near and/or inside the CU. The aim of this study was to verify whether P is the sperm chemoattractant secreted by the rabbit oocyte-cumulus complex (OCC) in the rabbit, as a mammalian animal model. By means of videomicroscopy and computer image analysis we observed that only the CU are a stable source of sperm attractants. The CU produce and secrete P since the hormone was localized inside these cells by immunocytochemistry and in the conditioned medium by enzyme immunoassay. In addition, rabbit spermatozoa express a cell surface P receptor detected by western blot and localized over the acrosomal region by immunocytochemistry. To confirm that P is the sperm chemoattractant secreted by the CU, the sperm chemotactic response towards the OCC conditioned medium was inhibited by three different approaches: P from the OCC conditioned medium was removed with an anti-P antibody, the attractant gradient of the OCC conditioned medium was disrupted by a P counter gradient, and the sperm P receptor was blocked with a specific antibody. We concluded that only the CU but not the oocyte secretes P, and the latter chemoattract spermatozoa by means of a cell surface receptor. Our findings may be of interest in assisted reproduction procedures in humans, animals of economic importance and endangered species. PMID:18725941

Localization of a small change in a multiple scattering environment without modeling of the actual medium.

PubMed

Rakotonarivo, S T; Walker, S C; Kuperman, W A; Roux, P

2011-12-01

A method to actively localize a small perturbation in a multiple scattering medium using a collection of remote acoustic sensors is presented. The approach requires only minimal modeling and no knowledge of the scatterer distribution and properties of the scattering medium and the perturbation. The medium is ensonified before and after a perturbation is introduced. The coherent difference between the measured signals then reveals all field components that have interacted with the perturbation. A simple single scatter filter (that ignores the presence of the medium scatterers) is matched to the earliest change of the coherent difference to localize the perturbation. Using a multi-source/receiver laboratory setup in air, the technique has been successfully tested with experimental data at frequencies varying from 30 to 60 kHz (wavelength ranging from 0.5 to 1 cm) for cm-scale scatterers in a scattering medium with a size two to five times bigger than its transport mean free path. © 2011 Acoustical Society of America

Diagnosing women with HPV: the impact of diagnosis disclosure methods.

PubMed

Harvey-Knowles, Jacquelyn A; Kosenko, Kami A

2012-07-01

Little data exists on women's HPV diagnosis encounters. This research focuses on impacts of the communicative medium used to inform women of their HPV status. We conducted a qualitative study to identify the mediums used to communicate HPV diagnoses and the impact of each medium on the diagnosis experience. Twenty-five women with HPV completed semi-structured interviews, which we recorded and transcribed. We relied on grounded theory techniques in both data collection and analysis. There are three primary mediums health care providers use to inform women of their HPV diagnosis: phone calls, mailed letters/email, and face-to-face interactions. Implications regarding each medium are identified and discussed. There are drawbacks associated with the use of each medium that healthcare practitioners should be aware of and seek to avoid. Healthcare providers can utilize descriptions of diagnosis encounters and the recommendations accompanying them to understand and modify ways they choose to inform individuals of an HPV diagnosis. Published by Elsevier Ireland Ltd.

Collective Beer Brand Identity: A Semiotic Analysis of the Websites Representing Small and Medium Enterprises in the Brewing Industry of Western PA

ERIC Educational Resources Information Center

Cincotta, Dominic

2014-01-01

This research studies how brand identities, individually and communally, as read through websites are created among small and medium-sized enterprise breweries in western Pennsylvania. Content analysis through the frame of Kress and van Leeuwen was used as the basis for the codebook that reads each brand identity for the researcher. The…

37 CFR Appendix B to Part 202 - “Best Edition” of Published Copyrighted Works for the Collections of the Library of Congress

Code of Federal Regulations, 2013 CFR

2013-07-01

... rather than less-permanent paper stock or printing process. III. Motion Pictures Film medium is considered a better quality than any other medium. The formats under “film” and “video formats” are listed in... works, editions with the greatest amount of information rather than those with less detail. C. Rarity: 1...

37 CFR Appendix B to Part 202 - “Best Edition” of Published Copyrighted Works for the Collections of the Library of Congress

Code of Federal Regulations, 2014 CFR

2014-07-01

... rather than less-permanent paper stock or printing process. III. Motion Pictures Film medium is considered a better quality than any other medium. The formats under “film” and “video formats” are listed in... works, editions with the greatest amount of information rather than those with less detail. C. Rarity: 1...

37 CFR Appendix B to Part 202 - “Best Edition” of Published Copyrighted Works for the Collections of the Library of Congress

Code of Federal Regulations, 2012 CFR

2012-07-01

... rather than less-permanent paper stock or printing process. III. Motion Pictures Film medium is considered a better quality than any other medium. The formats under “film” and “video formats” are listed in... works, editions with the greatest amount of information rather than those with less detail. C. Rarity: 1...

Patterns of Availability and Use of Audiotape Cassettes in Special Libraries. Ph.D. Thesis

NASA Technical Reports Server (NTRS)

Hughes, J. M., II

1975-01-01

The availability and use of audiotape cassettes is studied in terms of user requirements. The following factors were examined: how special libraries utilize audiotape cassettes; who the users of the medium are; how the libraries acquire and maintain their collection; and opinions of librarians as to the value of the audiotape cassette as a medium for dissemination of information.

Sialidase activities of cultured human fibroblasts and the metabolism of GM3 ganglioside

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

Usuki, S.; Lyu, S.C.; Sweeley, C.C.

1988-05-15

Free sialic acid has been found in the cell-conditioned medium of human foreskin fibroblasts. It is proposed that the accumulation of extracellular sialic acid may result from the hydrolysis of GM3 ganglioside on the cell surface of these fibroblasts. Sialidase activities with GM3 ganglioside and sialyllactitol as substrates were demonstrated in cell-conditioned medium, and the levels of their activities correlated positively with cell density. The GM3 sialidase activity at pH 4.5 was 4.1 and 38 pmol/h/ml of medium at sparse and confluent densities, respectively; the corresponding activities with sialyllactitol as the substrate were 12 and 75 pmol/h/ml of medium (pHmore » 4.5). The pH versus activity profiles with GM3 as the substrate suggested the presence of a second sialidase with an optimal activity at pH 6.5 in the conditioned medium of preconfluent cells. This activity was virtually absent in the medium of contact-inhibited cells and could not be assayed with sialyllactitol as the substrate. The turnover of cell surface GM3 was assessed by pulse labeling human foreskin fibroblasts with a radioactive precursor of sialic acid ((1-14C)N-acetylmannosamine) and a radioactive precursor of ceramide ((3,3-3H2)serine). During a chase period of 24 h turnover of the doubly labeled cellular GM3 was observed; there was a loss of about 35% of the 14C-labeled sialic acid without any measureable loss of 3H-labeled ceramide from GM3. We have speculated that the enzyme-catalyzed removal of sialic acid from the GM3 ganglioside on the extracellular aspect of the plasma membrane may be a necessary event involved in the modulation of cell growth.« less

[Marrow stromal cells cultured in N2-supplemented medium: implications on the generation of neural cells].

PubMed

Castillo-Díaz, L; de la Cuétara-Bernal, K; García-Varona, A Y

Most of the culture system for in vitro maintenance and neural differentiation of marrow stromal cells (MSCs) use synthetic media supplemented with 10 or 20% fetal bovine serum (FBS). Serum, however, is comprised of unknown quantities of undefined substances which could interfere the effect of exogenous substances on neural differentiation of MSCs. AIM. Here we describe survival of MSCs cultured in culture conditions where serum was reduced at 0.5 and 1% using Bottenstein and Sato's N2 formula (1979) and poly-L-lysine (PLL)-coated substrate. Stromal cells isolated from rat femurs were cultivated in Dulbecco's modified Eagle medium at 10, 1, 0.5% FBS or in serum free medium containing N2 formula. In serum free medium or at low serum concentration culture surface was coated with PLL. Cell survival was determined by MTT method or by counting viable cells. Survival of MSCs cultured in N2 supplement was reduced at about 40% of that observed in 10% FBS containing medium. Under these conditions cell morphology was also affected. When N2 containing medium was supplemented with FBS at 0.5 or 1% a significant increase of survival with respect to that observed in N2-supplemented cultures was observed. Cells seeded on PLL-coated surface increased their survival by contrast with their homologous cultures seeded on uncoated surface. The culture system which combines N2 formula with FBS 1% and PLL-coated surface is useful for the maintenance of MSCs. These conditions offer advantages for the study of differentiation of these cells because they reduce the confounding influence of serum. The possible implication of this culture system for the study of neural differentiation by these cells is discussed.

Na/Ca exchange in the basolateral membrane of the A6 cell monolayer: role in Cai homeostasis.

PubMed

Brochiero, E; Raschi, C; Ehrenfeld, J

1995-05-01

The presence of a Na/Ca exchanger in A6 cells was investigated by measuring intracellular calcium (Cai) fluctuations and the 45Ca fluxes through the basolateral membranes (blm) of the cell monolayer. Removal of Na+ from the medium produced a transient increase in Cai followed by a regulatory phase returning Cai to control levels in 3-4 min, this phase being greatly accelerated (< 60 s) by NaCl addition (apparent Km of approximately 5 mM Na+). The Cai increase was only found with the Na(+)-free medium on the basolateral side of the cell monolayer. A twofold increase in the 45Ca influx was observed under these conditions. In Ca(2+)- depleted cells, the initial Cai increase after Ca2+ addition to the medium was greater when the putative Na/Ca exchanger was not functioning (i.e. in a Na(+)-free medium). 45Ca effluxes through the blm of the monolayer were greatly and transiently increased by a Na(+)-free medium on the serosal side and blocked by orthovanadate (1 mM). The Cai increased induced by a hypo-osmotic shock was greater in cells bathed in a Na(+)-medium, conditions expected to block the activity of the Na/Ca exchanger. These findings support the hypothesis that a Na/Ca exchanger is present on the blm of A6 cells and affirm its role in Cai homeostasis in steady-state conditions and following osmotic shock. In addition, a Ca2+ pump also located on the blm and Ca2+ stores sensitive to inositol 1,4,5-trisphosphate were found to be implicated in Cai homeostasis.

Bank Size and Small- and Medium-sized Enterprise (SME) Lending: Evidence from China.

PubMed

Shen, Yan; Shen, Minggao; Xu, Zhong; Bai, Ying

2009-04-01

Using panel data collected in 2005, we evaluate how bank size, discretion over credit, incentive schemes, competition, and the institutional environment affect lending to small- and medium-sized enterprises in China. We deal with the endogeneity problem using instrumental variables, and a reduced-form approach is also applied to allow for weak instruments in estimation. We find that total bank asset is an insignificant factor for banks' decision on small- and medium-enterprise (SME) lending, but more local lending authority, more competition, carefully designed incentive schemes, and stronger law enforcement encourage commercial banks to lend to SMEs.

Steady-state heat transfer in transversely heated porous media with application to focused solar energy collectors

NASA Technical Reports Server (NTRS)

Nichols, L. D.

1976-01-01

A fluid flowing in a porous medium heated transversely to the fluid flow is considered. This configuration is applicable to a focused solar energy collector for use in an electric power generating system. A fluidized bed can be regarded as a porous medium with special properties. The solutions presented are valid for describing the effectiveness of such a fluidized bed for collecting concentrated solar energy to heat the working fluid of a heat engine. Results indicate the advantage of high thermal conductivity in the transverse direction and high operating temperature of the porous medium.

Blackbody emission from light interacting with an effective moving dispersive medium.

PubMed

Petev, M; Westerberg, N; Moss, D; Rubino, E; Rimoldi, C; Cacciatori, S L; Belgiorno, F; Faccio, D

2013-07-26

Intense laser pulses excite a nonlinear polarization response that may create an effective flowing medium and, under appropriate conditions, a blocking horizon for light. Here, we analyze in detail the interaction of light with such laser-induced flowing media, fully accounting for the medium dispersion properties. An analytical model based on a first Born approximation is found to be in excellent agreement with numerical simulations based on Maxwell's equations and shows that when a blocking horizon is formed, the stimulated medium scatters light with a blackbody emission spectrum. Based on these results, diamond is proposed as a promising candidate medium for future studies of Hawking emission from artificial, dispersive horizons.

Enhanced Oil Production by the Tropical Marine Diatom Thalassiosira Sp. Cultivated in Outdoor Photobioreactors.

PubMed

Kusumaningtyas, Pintaka; Nurbaiti, Santi; Suantika, Gede; Amran, Muhammad Bachri; Nurachman, Zeily

2017-08-01

Microalgae-derived oils have potential as a biofuel feedstock. To produce microalgal oils at a large scale, large amounts of nutrients and energy are needed to grow the algae. In this study, we evaluated three types of agricultural fertilizer (AF)-based culture media (AF1, AF2, and AF3) based on a previously published enriched seawater (ES) medium to produce biomass and oils from Thalassiosira sp. Under laboratory conditions, the highest cell productivity of Thalassiosira sp. was obtained with the AF3 medium. Thalassiosira sp. cultured in the AF3 medium produced 10.4 ± 0.9 mg L -1  day -1 oils, which is significantly higher than the 5.8 ± 0.7 mg L -1  day -1 produced in the ES medium. The higher production was due to the presence of nitrate and trace elements, both of which played roles in enhancing biomass and oil content, respectively. During cell growth, resting spores appeared inside the cells and were a marker to harvest the cells. Because of the abundant availability of sunlight in the tropics during the year, the oil production of Thalassiosira sp. in the AF3 medium was scaled up using outdoor photobioreactors under different weather conditions (rainy and dry seasons). Thalassiosira sp. produced more unsaturated fatty acids during the rainy season and produced more saturated fatty acids during the dry season. This study also demonstrated that it was possible to culture Thalassiosira sp. under outdoor conditions using a low-cost agricultural fertilizer-based culture medium (AF3 medium) to produce biodiesel feedstock with an annual production of 8.1 ± 0.4 t ha -1 during the dry season and of 23.9 ± 6.8 t ha -1 during the rainy season.

Flow-controlled magnetic particle manipulation

DOEpatents

Grate, Jay W [West Richland, WA; Bruckner-Lea, Cynthia J [Richland, WA; Holman, David A [Las Vegas, NV

2011-02-22

Inventive methods and apparatus are useful for collecting magnetic materials in one or more magnetic fields and resuspending the particles into a dispersion medium, and optionally repeating collection/resuspension one or more times in the same or a different medium, by controlling the direction and rate of fluid flow through a fluid flow path. The methods provide for contacting derivatized particles with test samples and reagents, removal of excess reagent, washing of magnetic material, and resuspension for analysis, among other uses. The methods are applicable to a wide variety of chemical and biological materials that are susceptible to magnetic labeling, including, for example, cells, viruses, oligonucleotides, proteins, hormones, receptor-ligand complexes, environmental contaminants and the like.

Modeling the VARTM Composite Manufacturing Process

NASA Technical Reports Server (NTRS)

Song, Xiao-Lan; Loos, Alfred C.; Grimsley, Brian W.; Cano, Roberto J.; Hubert, Pascal

2004-01-01

A comprehensive simulation model of the Vacuum Assisted Resin Transfer Modeling (VARTM) composite manufacturing process has been developed. For isothermal resin infiltration, the model incorporates submodels which describe cure of the resin and changes in resin viscosity due to cure, resin flow through the reinforcement preform and distribution medium and compaction of the preform during the infiltration. The accuracy of the model was validated by measuring the flow patterns during resin infiltration of flat preforms. The modeling software was used to evaluate the effects of the distribution medium on resin infiltration of a flat preform. Different distribution medium configurations were examined using the model and the results were compared with data collected during resin infiltration of a carbon fabric preform. The results of the simulations show that the approach used to model the distribution medium can significantly effect the predicted resin infiltration times. Resin infiltration into the preform can be accurately predicted only when the distribution medium is modeled correctly.

Modeling the efficiency of a magnetic needle for collecting magnetic cells

NASA Astrophysics Data System (ADS)

Butler, Kimberly S.; Adolphi, Natalie L.; Bryant, H. C.; Lovato, Debbie M.; Larson, Richard S.; Flynn, Edward R.

2014-07-01

As new magnetic nanoparticle-based technologies are developed and new target cells are identified, there is a critical need to understand the features important for magnetic isolation of specific cells in fluids, an increasingly important tool in disease research and diagnosis. To investigate magnetic cell collection, cell-sized spherical microparticles, coated with superparamagnetic nanoparticles, were suspended in (1) glycerine-water solutions, chosen to approximate the range of viscosities of bone marrow, and (2) water in which 3, 5, 10 and 100% of the total suspended microspheres are coated with magnetic nanoparticles, to model collection of rare magnetic nanoparticle-coated cells from a mixture of cells in a fluid. The magnetic microspheres were collected on a magnetic needle, and we demonstrate that the collection efficiency versus time can be modeled using a simple, heuristically-derived function, with three physically-significant parameters. The function enables experimentally-obtained collection efficiencies to be scaled to extract the effective drag of the suspending medium. The results of this analysis demonstrate that the effective drag scales linearly with fluid viscosity, as expected. Surprisingly, increasing the number of non-magnetic microspheres in the suspending fluid results increases the collection of magnetic microspheres, corresponding to a decrease in the effective drag of the medium.

Modeling the Efficiency of a Magnetic Needle for Collecting Magnetic Cells

PubMed Central

Butler, Kimberly S; Adolphi, Natalie L.; Bryant, H C; Lovato, Debbie M; Larson, Richard S; Flynn, Edward R

2014-01-01

As new magnetic nanoparticle-based technologies are developed and new target cells are identified, there is a critical need to understand the features important for magnetic isolation of specific cells in fluids, an increasingly important tool in disease research and diagnosis. To investigate magnetic cell collection, cell-sized spherical microparticles, coated with superparamagnetic nanoparticles, were suspended in 1) glycerine-water solutions, chosen to approximate the range of viscosities of bone marrow, and 2) water in which 3, 5, 10 and 100 % of the total suspended microspheres are coated with magnetic nanoparticles, to model collection of rare magnetic nanoparticle-coated cells from a mixture of cells in a fluid. The magnetic microspheres were collected on a magnetic needle, and we demonstrate that the collection efficiency vs. time can be modeled using a simple, heuristically-derived function, with three physically-significant parameters. The function enables experimentally-obtained collection efficiencies to be scaled to extract the effective drag of the suspending medium. The results of this analysis demonstrate that the effective drag scales linearly with fluid viscosity, as expected. Surprisingly, increasing the number of non-magnetic microspheres in the suspending fluid results increases the collection of magnetic microspheres, corresponding to a decrease in the effective drag of the medium. PMID:24874577

Establishment of an effective oligotrophic cultivation system for Rhodococcus erythropolis N9T-4.

PubMed

Matsuoka, Tomohiro; Yoshida, Nobuyuki

2018-06-03

Rhodococcus erythropolis N9T-4 grows on an inorganic solid-state medium with no additional carbon and energy sources; however, it is unable to grow well in a liquid culture medium under the oligotrophic conditions. We examined submerged cultivations of N9T-4 using a polyurethane foam sponge to achieve approximately 10 times of the oligotrophic growth of the bacterium in the liquid culture medium.

Optimization of the Synthesis of Structured Phosphatidylcholine with Medium Chain Fatty Acid.

PubMed

Ochoa-Flores, Angélica A; Hernández-Becerra, Josafat A; Cavazos-Garduño, Adriana; Vernon-Carter, Eduardo J; García, Hugo S

2017-11-01

Structured phosphatidylcholine was successfully produced by acidolysis between phosphatidylcholine and free medium chain fatty acid, using phospholipase A 1 immobilized on Duolite A568. Response surface methodology was applied to optimize the reaction system using three process parameters: molar ratio of substrates (phosphatidylcholine to free medium chain fatty acid), enzyme loading, and reaction temperature. All parameters evaluated showed linear and quadratic significant effects on the production of modified phosphatidylcholine; molar ratio of substrates contributed positively, but temperature influenced negatively. Increased enzyme loading also led to increased production of modified phosphatidylcholine but only during the first 9 hours of the acidolysis reaction. Optimal conditions obtained from the model were a ratio of phosphatidylcholine to free medium chain fatty acid of 1:15, an enzyme loading of 12%, and a temperature of 45°C. Under these conditions a production of modified phosphatidylcholine of 52.98 % were obtained after 24 h of reaction. The prediction was confirmed from the verification experiments; the production of modified phosphatidylcholine was 53.02%, the total yield of phosphatidylcholine 64.28% and the molar incorporation of medium chain fatty acid was 42.31%. The acidolysis reaction was scaled-up in a batch reactor with a similar production of modified phosphatidylcholine, total yield of phosphatidylcholine and molar incorporation of medium chain fatty acid. Purification by column chromatography of the structured phosphatidylcholine yielded 62.53% of phosphatidylcholine enriched with 42.52% of medium chain fatty acid.

Powertrain Test Procedure Development for EPA GHG Certification of Medium- and Heavy-Duty Engines and Vehicles

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

Chambon, Paul H.; Deter, Dean D.

2016-07-01

xiii ABSTRACT The goal of this project is to develop and evaluate powertrain test procedures that can accurately simulate real-world operating conditions, and to determine greenhouse gas (GHG) emissions of advanced medium- and heavy-duty engine and vehicle technologies. ORNL used their Vehicle System Integration Laboratory to evaluate test procedures on a stand-alone engine as well as two powertrains. Those components where subjected to various drive cycles and vehicle conditions to evaluate the validity of the results over a broad range of test conditions. Overall, more than 1000 tests were performed. The data are compiled and analyzed in this report.

Using straw hydrolysate to cultivate Chlorella pyrenoidosa for high-value biomass production and the nitrogen regulation for biomass composition.

PubMed

Zhang, Tian-Yuan; Wang, Xiao-Xiong; Wu, Yin-Hu; Wang, Jing-Han; Deantes-Espinosa, Victor M; Zhuang, Lin-Lan; Hu, Hong-Ying; Wu, Guang-Xue

2017-11-01

Heterotrophic cultivation of Chlorella pyrenoidosa based on straw substrate was proposed as a promising approach in this research. The straw pre-treated by ammonium sulfite method was enzymatically hydrolyzed for medium preparation. The highest intrinsic growth rate of C. pyrenoidosa reached to 0.097h -1 in hydrolysate medium, which was quicker than that in glucose medium. Rising nitrogen concentration could significantly increase protein content and decrease lipid content in biomass, meanwhile fatty acids composition kept stable. The highest protein and lipid content in microalgal biomass reached to 62% and 32% under nitrogen excessive and deficient conditions, respectively. Over 40% of amino acids and fatty acids in biomass belonged to essential amino acids (EAA) and essential fatty acids (EFA), which were qualified for high-value uses. This research revealed the rapid biomass accumulation property of C. pyrenoidosa in straw hydrolysate medium and the effectiveness of nitrogen regulation to biomass composition at heterotrophic condition. Copyright © 2017. Published by Elsevier Ltd.

Effects of culture conditions on growth and docosahexaenoic acid production from Schizochytrium limacinum

NASA Astrophysics Data System (ADS)

Zhu, Luying; Zhang, Xuecheng; Ren, Xueying; Zhu, Qinghua

2008-02-01

The effects of temperature, initial pH, salinity of culture medium, and carbon and nitrogen sources on growth and docosahexaenoic acid (C22: 6 n-3, DHA) production from Schizochytrium limacinum OUC88 were investigated in the present study. The results revealed that the optimal temperature, initial pH and salinity level of the medium for DHA production were 23°C, 7.0 and 18, respectively. Glucose was proved the best carbon source for the growth and DHA production from S. limacinum. Among the nitrogen sources tested, soybean cake hydrolysate, a cheap by-product, was found to be effective for the accumulation of DHA in S. limacinum cells. In addition, increasing the concentration of carbon sources in the medium caused a significant increase in cell biomass; however, accumulation of DHA in cells was mainly stimulated by the ratio of C/N in the medium. Under the optimal culture conditions, the maximum DHA yield achieved in flasks was 4.08 g L-1 after 5 d of cultivation.

A novel steam explosion sterilization improving solid-state fermentation performance.

PubMed

Zhao, Zhi-Min; Wang, Lan; Chen, Hong-Zhang

2015-09-01

Traditional sterilization of solid medium (SM) requires lengthy time, degrades nutrients, and even sterilizes inadequately compared with that of liquid medium due to its low thermal conductivity. A novel sterilization strategy, high-temperature and short-time steam explosion (SE), was exploited for SM sterilization in this study. Results showed that SE conditions for complete sterilization were 172 °C for 2 min and 128 °C for 5 min. Glucose and xylose contents in medium after SE sterilization increased by 157% and 93% respectively compared with those after conventional sterilization (121 °C, 20 min) while fermentation inhibitors were not detected. FTIR spectra revealed that the mild SE conditions helped to release monosaccharides from the polysaccharides. Bacillus subtilis fermentation productivity on medium after SE sterilization was 3.83 times of that after conventional sterilization. Therefore, SE shortened sterilization time and improved SM nutrition, which facilitated fermentability of SM and should promote economy of solid-state fermentation. Copyright © 2015 Elsevier Ltd. All rights reserved.

Image analysis technique as a tool to identify morphological changes in Trametes versicolor pellets according to exopolysaccharide or laccase production.

PubMed

Tavares, Ana P M; Silva, Rui P; Amaral, António L; Ferreira, Eugénio C; Xavier, Ana M R B

2014-02-01

Image analysis technique was applied to identify morphological changes of pellets from white-rot fungus Trametes versicolor on agitated submerged cultures during the production of exopolysaccharide (EPS) or ligninolytic enzymes. Batch tests with four different experimental conditions were carried out. Two different culture media were used, namely yeast medium or Trametes defined medium and the addition of lignolytic inducers as xylidine or pulp and paper industrial effluent were evaluated. Laccase activity, EPS production, and final biomass contents were determined for batch assays and the pellets morphology was assessed by image analysis techniques. The obtained data allowed establishing the choice of the metabolic pathways according to the experimental conditions, either for laccase enzymatic production in the Trametes defined medium, or for EPS production in the rich Yeast Medium experiments. Furthermore, the image processing and analysis methodology allowed for a better comprehension of the physiological phenomena with respect to the corresponding pellets morphological stages.

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.

Evaluation of the FTA carrier device for human papillomavirus testing in developing countries.

PubMed

Gonzalez, Paula; Cortes, Bernal; Quint, Wim; Kreimer, Aimée R; Porras, Carolina; Rodríguez, Ana Cecilia; Jimenez, Silvia; Herrero, Rolando; Struijk, Linda; Hildesheim, Allan; Melchers, Willem

2012-12-01

Liquid-based methods for the collection, transportation, and storage of cervical cells are cumbersome and expensive and involve laborious DNA extraction. An FTA cartridge is a solid carrier device, easier to handle and allowing simple DNA elution for human papillomavirus (HPV) testing. HPV-DNA results from cervical specimens collected in PreservCyt medium (Hologic, Inc.) and the indicating FTA elute cartridge were compared in an area where transportation and storage may affect the performance of the test. Cervical cells from 319 young adult women enrolled in the Costa Rica Vaccine Trial were collected by a nurse using a Cervex brush (Roberts), which was placed on the FTA cartridge and subsequently rinsed in 20 ml of PreservCyt medium. Two 0.5-ml PreservCyt aliquots were frozen for HPV-PCR testing; the FTA cartridges were kept at room temperature. HPV-DNA detection and typing was performed using SPF(10) PCR/DEIA (DNA enzyme immunoassay detection of amplimers)/LiPA(25) system. The percent agreement, agreement among positives, and kappas were estimated. Positivity was higher for FTA compared to PreservCyt specimens (54.5% versus 45.8%, P < 0.001). For oncogenic types, the overall agreement was 0.92, the agreement between positives was 0.74, and the kappa was 0.79. For individual HPV types, the overall agreement ranged from 0.97 to 1.00. We did not observe reduced cytology adequacy when specimen collection for cytology was preceded by FTA collection for HPV testing. HPV-DNA detection from FTA cartridges is broadly comparable to detection from PC medium. The higher HPV detection observed for FTA-collected specimens should be explored further. FTA cartridges could provide a simpler and more cost-effective method for cervical cell collection, storage, and transportation for HPV-DNA detection in research settings in developing countries.

Evaluation of the FTA Carrier Device for Human Papillomavirus Testing in Developing Countries

PubMed Central

Cortes, Bernal; Quint, Wim; Kreimer, Aimée R.; Porras, Carolina; Rodríguez, Ana Cecilia; Jimenez, Silvia; Herrero, Rolando; Struijk, Linda; Hildesheim, Allan; Melchers, Willem

2012-01-01

Liquid-based methods for the collection, transportation, and storage of cervical cells are cumbersome and expensive and involve laborious DNA extraction. An FTA cartridge is a solid carrier device, easier to handle and allowing simple DNA elution for human papillomavirus (HPV) testing. HPV-DNA results from cervical specimens collected in PreservCyt medium (Hologic, Inc.) and the indicating FTA elute cartridge were compared in an area where transportation and storage may affect the performance of the test. Cervical cells from 319 young adult women enrolled in the Costa Rica Vaccine Trial were collected by a nurse using a Cervex brush (Roberts), which was placed on the FTA cartridge and subsequently rinsed in 20 ml of PreservCyt medium. Two 0.5-ml PreservCyt aliquots were frozen for HPV-PCR testing; the FTA cartridges were kept at room temperature. HPV-DNA detection and typing was performed using SPF10 PCR/DEIA (DNA enzyme immunoassay detection of amplimers)/LiPA25 system. The percent agreement, agreement among positives, and kappas were estimated. Positivity was higher for FTA compared to PreservCyt specimens (54.5% versus 45.8%, P < 0.001). For oncogenic types, the overall agreement was 0.92, the agreement between positives was 0.74, and the kappa was 0.79. For individual HPV types, the overall agreement ranged from 0.97 to 1.00. We did not observe reduced cytology adequacy when specimen collection for cytology was preceded by FTA collection for HPV testing. HPV-DNA detection from FTA cartridges is broadly comparable to detection from PC medium. The higher HPV detection observed for FTA-collected specimens should be explored further. FTA cartridges could provide a simpler and more cost-effective method for cervical cell collection, storage, and transportation for HPV-DNA detection in research settings in developing countries. PMID:22993174

Prediction of fingering in porous media

NASA Astrophysics Data System (ADS)

Wang, Zhi; Feyen, Jan; Elrick, David E.

1998-09-01

Immiscible displacement, involving two fluids in a porous medium, can be unstable and fingered under certain conditions. In this paper, the original linear instability criterion of Chuoke et al. [1959] is generalized, considering wettability of two immiscible fluids to the porous medium. This is then used to predict 24 specific flow and porous medium conditions for the onset of wetting front instability in the subsurface. Wetting front instability is shown to be a function of the driving fluid wettability to the medium, differences in density and viscosity of the fluids, the magnitude of the interfacial tension, and the direction of flow with respect to gravity. Scenarios of water and nonaqueous-phase liquid infiltration into the vadose zone are examined to predict preferential flow and contamination of groundwater. The mechanisms of finger formation, propagation, and persistence in the vadose zone are reviewed, and the existing equations for calculating the size, the number and velocity of fingers are simplified for field applications. The analyses indicate that fingers initiate and propagate according to spatial and temporal distribution of the dynamic breakthrough (water- or air-entry) pressures in the porous medium. The predicted finger size and velocity are in close agreement with the experimental results.

New plant-growth medium for increased power output of the Plant-Microbial Fuel Cell.

PubMed

Helder, M; Strik, D P B T B; Hamelers, H V M; Kuijken, R C P; Buisman, C J N

2012-01-01

In a Plant-Microbial Fuel Cell anode-conditions must be created that are favorable for plant growth and electricity production. One of the major aspects in this is the composition of the plant-growth medium. Hoagland medium has been used until now, with added phosphate buffer to reduce potential losses over the membrane because of differences in pH between anode and cathode. We developed a new, improved plant-growth medium that improves current production, while the plant keeps growing. This medium is a nitrate-less, ammonium-rich medium that contains all macro- and micro-nutrients necessary for plant growth, with a balanced amount of bicarbonate buffer. Sulphate presence in the plant-growth medium helps to keep a low anode-potential. With the new plant-growth medium the maximum current production of the Plant-Microbial Fuel Cell increased from 186 mA/m(2) to 469 mA/m(2). Copyright © 2011 Elsevier Ltd. All rights reserved.

Growth medium alterations improve in vitro cold storage of pear germplasm

USDA-ARS?s Scientific Manuscript database

Development of new fruit cultivars is dependent on genetic resource collections such as those at the Pomological Garden at the Institute of Horticulture and Viticulture near Almaty, Kazakhstan. The pear germplasm collection of the Pomological Garden contains 615 cultivars and 3 species. About 36 ac...

Data Collection from Videotaped Experiments in a Biology Laboratory

ERIC Educational Resources Information Center

Russock, Howard I.

1977-01-01

Describes individualized television instruction involving active student participation, where experiments on air breathing in fish and shoot growth were videotaped. Students collected data directly from the television screen. Student accuracy was found to be excellent; reactions to the medium were found to be apathetic. Reasons for apathy are…

Bromelain enzyme from pineapple: in vitro activity study under different micropropagation conditions.

PubMed

Vilanova Neta, Jaci Lima; da Silva Lédo, Ana; Lima, Aloisio André Bonfim; Santana, José Carlos Curvelo; Leite, Nadjma Souza; Ruzene, Denise Santos; Silva, Daniel Pereira; de Souza, Roberto Rodrigues

2012-09-01

The aim of this work was to evaluate the activity of bromelain in pineapple plants (Ananas comosus var. Comosus), Pérola cultivar, produced in vitro in different culture conditions. This enzyme, besides its pharmacological effects, is also employed in food industries, such as breweries and meat processing. In this work, the enzymatic activity was evaluated in the tissues of leaves and stems of plants grown in culture medium without plant growth regulator. The most significant levels of bromelain were observed in leaf tissue after 4 months of culture in vitro in medium with a filter paper bridge, followed by medium gelled by the agar. The results of this study, regarding the different structures of the pineapple (leaves and stems) in vitro showed that the activity of bromelain varied depending on the culture conditions, the time and structure of which was quantified, ensuring a viable strategy in the production of seedlings with high levels of bromelain in subsequent phases of micropropagation.

Collapsing Aged Culture of the Cyanobacterium Synechococcus elongatus Produces Compound(s) Toxic to Photosynthetic Organisms

PubMed Central

Cohen, Assaf; Sendersky, Eleonora; Carmeli, Shmuel; Schwarz, Rakefet

2014-01-01

Phytoplankton mortality allows effective nutrient cycling, and thus plays a pivotal role in driving biogeochemical cycles. A growing body of literature demonstrates the involvement of regulated death programs in the abrupt collapse of phytoplankton populations, and particularly implicates processes that exhibit characteristics of metazoan programmed cell death. Here, we report that the cell-free, extracellular fluid (conditioned medium) of a collapsing aged culture of the cyanobacterium Synechococcus elongatus is toxic to exponentially growing cells of this cyanobacterium, as well as to a large variety of photosynthetic organisms, but not to eubacteria. The toxic effect, which is light-dependent, involves oxidative stress, as suggested by damage alleviation by antioxidants, and the very high sensitivity of a catalase-mutant to the conditioned medium. At relatively high cell densities, S. elongatus cells survived the deleterious effect of conditioned medium in a process that required de novo protein synthesis. Application of conditioned medium from a collapsing culture caused severe pigment bleaching not only in S. elongatus cells, but also resulted in bleaching of pigments in a cell free extract. The latter observation indicates that the elicited damage is a direct effect that does not require an intact cell, and therefore, is mechanistically different from the metazoan-like programmed cell death described for phytoplankton. We suggest that S. elongatus in aged cultures are triggered to produce a toxic compound, and thus, this process may be envisaged as a novel regulated death program. PMID:24959874

Properties of Polyhydroxyalkanoate Granules and Bioemulsifiers from Pseudomonas sp. and Burkholderia sp. Isolates Growing on Glucose.

PubMed

Sacco, Laís Postai; Castellane, Tereza Cristina Luque; Lopes, Erica Mendes; de Macedo Lemos, Eliana Gertrudes; Alves, Lúcia Maria Carareto

2016-03-01

A Burkholderia and Pseudomonas species designated as AB4 and AS1, respectively, were isolated from soil containing decomposing straw or sugar cane bagasse collected from Brazil. This study sought to evaluate the capacities of culture media, cell-free medium, and crude lysate preparations (containing PHB inclusion bodies) from bacterial cell cultures to stabilize emulsions with several hydrophobic compounds. Four conditions showed good production of bioemulsifiers (E24 ≥ 50 %), headed by substantially cell-free media from bacterial cell cultures in which bacterial isolates from Burkholderia sp. strain AB4 and Pseudomonas sp. strain AS1 were grown. Our results revealed that the both isolates (AB4 and AS1 strains) exhibited high emulsification indices (indicating usefulness in bioremediation) and good stabilities.

Optimization of culturing condition and medium composition for the production of alginate lyase by a marine Vibrio sp. YKW-34

NASA Astrophysics Data System (ADS)

Fu, Xiaoting; Lin, Hong; Kim, Sang Moo

2008-02-01

Carbohydrases secreted by marine Vibrio sp. YKW-34 with strong Laminaria cell wall degrading ability were screened, and among them alginate lyase was found to be dominant. The effects of medium composition and culturing condition on the production of alginate lyase by marine Vibrio sp. YKW-34 in flask were investigated in this study. In the culture medium of marine broth, no alginate lyase was produced. The activity of the alginate lyase, after being induced, reached 5 UmL-1. The best inoculum volume and inoculum age were 10% and 12 h, respectively. The optimal temperature for alginate lyase production was 25°C. The fermentation medium was composed of 0.5% of Laminaria powder and 0.2% of KNO3 with an initial acidity of pH 8.0. Alginate could induce alginate lyase production but not as efficiently as Laminaria powder did. The addition of fucoidan, cellulose and glucose had negative effect on the alginate lyase production. Other kinds of nitrogen sources, such as yeast extract, beef extract and peptone, had positive effect on the growth of the microorganism and negative effect on alginate lyase production. In addition, the time course of alginate lyase production under the optimized condition was described. The optimal harvest time was 48 h.

Microwave Signatures of Melting/Refreezing Snow: Observations and Modeling Using Dense Medium Radiative Transfer Theory

NASA Technical Reports Server (NTRS)

Tedesco, Marco; Kim, Edward J.; England, Anthony; deRoo, Roger; Hardy, Janet

2005-01-01

Microwave brightness temperatures of snow covered terrains can be modeled by means of the Dense Radiative Transfer Medium Theory (DMRT). In a dense medium, such as snow, the assumption of independent scattering is no longer valid and the scattering of correlated scatterers must be considered. In the DMRT, this is done considering a pair distribution function of the particles position. In the electromagnetic model, the snowpack is simulated as a homogeneous layer having effective permittivity and albedo calculated through the DMRT. In order to account for clustering of snow crystals, a model of cohesive particles can be applied, where the cohesion between the particles is described by means of a dimensionless parameters called stickiness (z), representing a measure of the inversion of the attraction of the particles. The lower the z the higher the stickiness. In this study, microwave signatures of melting and refreezing cycles of seasonal snowpacks at high altitudes are studied by means of both experimental and modeling tools. Radiometric data were collected 24 hours per day by the University of Michigan Tower Mounted Radiometer System (TMRS). The brightness temperatures collected by means of the TMRS are simulated by means of a multi-layer electromagnetic model based on the dense medium theory with the inputs to the model derived from the data collected at the snow pits and from the meteorological station. The paper is structured as follows: in the first Section the temperature profiles recorded by the meteorological station and the snow pit data are presented and analyzed; in the second Section, the characteristics of the radiometric system used to collect the brightness temperatures are reported together with the temporal behavior of the recorded brightness temperatures; in the successive Section the multi-layer DMRT-based electromagnetic model is described; in the fourth Section the comparison between modeled and measured brightness temperatures is discussed. We dedicate the last Section to the conclusions and future works.

Novel Preclinical Testing Strategies for Treatment of Metastatic Pheochromocytoma

DTIC Science & Technology

2013-09-01

proliferation using this protocol as reported or with any modifications tested Medium Y27632 (uM) Hydrocortisone (ug/mL) Insulin (ug/mL) rhEGF...0 + + 0 + + 0 + 0 Medium Y27632 (uM) Hydrocortisone (ug/mL) Insulin (ug/mL) rhEGF (ng/mL) Adenine (ug/mL) Result...reported (1) except for cholera toxin, which was toxic to this tumor under these conditions 17 Medium Y27632 (uM) Hydrocortisone (ug/mL

Kinetics of the Active Medium of a Copper Vapor Brightness Amplifier

NASA Astrophysics Data System (ADS)

Kulagin, A. E.; Torgaev, S. N.; Evtushenko, G. S.; Trigub, M. V.

2018-03-01

A spatiotemporal kinetics of the active medium of a copper vapor brightness amplifier is described that allows gain characteristics to be investigated during the pump pulse. Model calculations show that changing the discharge parameters allows the radial gain profiles to be improved significantly, as well as the gain and the inversion duration to be increased. The data obtained will be used to choose the operating conditions for the active medium in the brightness amplifier mode.

Antioxidant and neuronal cell protective effects of columbia arabica coffee with different roasting conditions.

PubMed

Jeong, Ji Hee; Jeong, Hee Rok; Jo, Yu Na; Kim, Hyun Ju; Lee, Uk; Heo, Ho Jin

2013-03-01

In vitro antioxidant activities and neuronal cell protective effects of ethanol extract from roasted coffee beans were investigated. Colombia arabica coffee (Coffea arabica) green beans were roasted to give medium (230°C, 10 min), city (230°C, 12 min) and french (230°C, 15 min) coffee beans. Total phenolics in raw green beans, medium, city and french-roasted beans were 8.81±0.05, 9.77±0.03, 9.92±0.04 and 7.76±0.01 mg of GAE/g, respectively. The content of 5-O-caffeoylquinic acid, the predominant phenolic, was detected higher in medium-roasted beans than others. In addition, we found that extracts from medium-roasted beans particularly showed the highest in vitro antioxidant activity on ABTS radical scavenging activity and FRAP assays. To determine cell viability using the MTT assay, extracts from medium-roasted beans showed higher protection against H2O2-induced neurotoxicity than others. Lactate dehydrogenase (LDH) leakage was also inhibited by the extracts due to prevention of lipid peroxidation using the malondialdehyde (MDA) assay from mouse whole brain homogenates. These data suggest that the medium-roasting condition to making tasty coffee from Columbia arabica green beans may be more helpful to human health by providing the most physiological phenolics, including 5-O-caffeoylquinic acids.

Propagation of monochromatic light in a hot and dense medium

NASA Astrophysics Data System (ADS)

Masood, Samina S.

2017-12-01

Photons, as quanta of electromagnetic fields, determine the electromagnetic properties of an extremely hot and dense medium. Considering the properties of the photons in the interacting medium of charged particles, we explicitly calculate the electromagnetic properties such as the electric permittivity, magnetic permeability, refractive index and the propagation speed of electromagnetic signals in an extremely hot and dense background. Photons acquire a dynamically generated mass in such a medium. The screening mass of the photon, the Debye shielding length and the plasma frequency are calculated as functions of the statistical parameters of the medium. We study the properties of the propagating particles in astrophysical systems of distinct statistical conditions. The modifications in the properties of the medium lead to the equation of state of the system. We mainly calculate all these parameters for extremely high temperatures of the early universe.

Transmission versus reflectance spectroscopy for quantitation

NASA Astrophysics Data System (ADS)

Gardner, Craig M.

2018-01-01

The objective of this work was to compare the accuracy of analyte concentration estimation when using transmission versus diffuse reflectance spectroscopy of a scattering medium. Monte Carlo ray tracing of light through the medium was used in conjunction with pure component absorption spectra and Beer-Lambert absorption along each ray's pathlength to generate matched sets of pseudoabsorbance spectra, containing water and six analytes present in skin. PLS regression models revealed an improvement in accuracy when using transmission compared to reflectance for a range of medium thicknesses and instrument noise levels. An analytical expression revealed the source of the accuracy degradation with reflectance was due both to the reduced collection efficiency for a fixed instrument etendue and to the broad pathlength distribution that detected light travels in the medium before exiting from the incident side.

Osmotic characteristics and fertility of murine spermatozoa collected in different solutions.

PubMed

Si, Wei; Men, Hongsheng; Benson, James D; Critser, John K

2009-02-01

Osmotic stress is an important factor that can result in cell damage during cryopreservation. Before ejaculation or collection for cryopreservation, murine spermatozoa are stored in epididymal fluid, a physiologically hyperosmotic environment (approximately 415 mmol/kg). The objectives of this study were to determine the osmotic tolerance limits of sperm motion parameters of ICR and C57BL/6 mouse spermatozoa collected in isosmotic (290 mmol/kg) and hyperosmotic (415 mmol/kg) media, and the effect of the osmolality of sperm collection media on sperm fertility after cryopreservation. Our results indicate that murine spermatozoa collected in media with different osmolalities (290 and 415 mmol/kg Dulbecco's phosphate buffered saline (DPBS)) appeared to have different osmotic tolerances for the maintenance of sperm motility and other motion parameters in both mouse strains. The hypo- and hyperosmotic treatments decreased motility and affected other motion parameters of spermatozoa collected in 290 mmol/kg DPBS. The extent of the change of motion parameters after treatments corresponded with the levels of osmotic stress. However, for spermatozoa collected in 415 mmol/kg DPBS, exposure to 290 mmol/kg DPBS tended to increase sperm motility and the quality of their motion parameters. The osmolality of sperm collection medium can affect murine sperm fertility. Spermatozoa collected in 415 mmol/kg medium showed higher fertility compared with spermatozoa collected in 290 mmol/kg as assessed by IVF. Results characterizing murine sperm osmotic tolerance collected in media with different osmolalities from different strains and the effect of collection media osmolality on sperm fertility after cryopreservation will be useful in designing cryopreservation protocols.

Higher production of C-phycocyanin by nitrogen-free (diazotrophic) cultivation of Nostoc sp. NK and simplified extraction by dark-cold shock.

PubMed

Lee, Na Kyeong; Oh, Hee-Mock; Kim, Hee-Sik; Ahn, Chi-Yong

2017-03-01

Nostoc sp. NK (KCTC 12772BP) was isolated and cultivated in a BG11 medium and a nitrate-free BG11 medium (BG11 0 ). To enhance C-phycocyanin (C-PC) content in the cells, different fluorescent lamps (white, plant, and red) were used as light sources for complementary chromatic adaptation (CCA). The maximum biomass productivity was 0.42g/L/d and 0.32g/L/d under BG11 and BG11 0 conditions, respectively. The maximum C-PC contents were 8.4% (w/w) under white lamps, 13.6% (w/w) under plant lamps, and 18% (w/w) under BG11 0 and the red light condition. The maximum C-PC productivity was 57.4mg/L/d in BG11 0 under the red lamp condition. These results indicate that a higher C-PC content could be obtained under a diazotrophic condition and a CCA reaction. The C-PC could be released naturally from cells without any extraction processes, when Nostoc sp. NK was cultivated in the BG11 0 medium with CO 2 aeration and put in dark conditions at 5°C. Copyright © 2016 Elsevier Ltd. All rights reserved.

Culture of porcine embryonic germ cells in serum-supplemented and serum-free conditions: the effects of serum and growth factors on primary and long-term culture.

PubMed

Petkov, Stoyan G; Anderson, Gary B

2008-06-01

Fetal bovine serum (FBS) is a commonly used medium supplement with variable and undefined composition, which presents problems in culture of pluripotent stem cells. The purpose of this study was to determine if FBS can be replaced with Knockout Serum Replacement (KSR), a defined medium supplement, and to examine the effects of FBS and growth factors on short- and long-term culture of pig embryonic germ cells (EGC). No significant differences were observed in total and mean colony areas in primary cultures between FBS- and KSR-supplemented medium (421 x 10(3) mum(2) vs. 395 x 10(3) microm(2), p = 0.68, n = 11, and 6375 microm(2) vs. 6407 microm(2), p = 0.885, respectively). Total and mean colony areas were significantly larger in KSR-supplemented medium compared with medium supplemented with KSR and growth factors (505 x 10(3) microm(2) vs. 396 x 10(3) microm(2), p = 0.016, n = 12, and 8769 microm(2) vs. 6513 microm(2), p = 0.003, respectively). The cultures proliferated for significantly higher numbers of passages in FBS-supplemented medium and in medium supplemented with KSR and growth factors compared with medium containing KSR alone (31.1 vs. 21.9, p = 0.004, n = 10, and 35.5 vs. 21.6, p = 002, n = 10, respectively). Porcine EGC maintained in serum-free conditions were positive for pluripotent stem cell markers, maintained stable karyotypes for up to 54 passages, and were capable of differentiating in vitro into cells from the three primary germ layers. These results will help improve and standardize culture of pluripotent stem cells in the pig.

Changes of vitamins A, E, and C and lipid peroxidation status of breeding and pregnant sheep during dry seasons on medium-to-low quality forages.

PubMed

Mohebbi-Fani, Mehdi; Mirzaei, Abdollah; Nazifi, Saeed; Shabbooie, Zahra

2012-02-01

This study assessed the changes of plasma vitamin A, E, and C and the lipid peroxidation status of sheep during breeding and pregnancy under drought conditions. The study was conducted on 105 cross-bred fat tailed ewes, 3-5 years old with body condition scores (BCS) of 2.5 to 3.5. The ewes were grazing on medium-to-low quality forages during summer and low quality forages within the succeeding months and had ad libitum access to a mixture of alfalfa hay (40%) and wheat straw (60%) in the afternoons. From 3 weeks before breeding till 1 month after the introduction of rams, 300 g of barley grain/head/day was offered to the ewes and then the supplemental grain was reduced to 100 g/head/day. For better synchronization of estrus cycles in ewes, they were isolated from the rams for at least 2 months and then kept in close proximity of the rams for 1 week before the introduction of the rams to the ewe flock. Then, whole blood samples were collected on days 1, 7, 21, and 120 after ram introduction. Vitamins A, E, and C were measured in plasma. Malondialdehyde (MDA) was measured in the hemolysate as a biomarker of lipid peroxidation. Plasma progesterone (P4) was measured in the samples of day 120 for assessing pregnancy status of the ewes. Vitamins A and C showed continuous and significant declines (P < 0.05) through days 1 to 120. Vitamin E declined only during the first 21 days of the study and remained almost constant till day 120. MDA concentration increased significantly at day 21. An elevated concentration of MDA was also detected at day 120. The difference between days 21 and 120 was not significant (P > 0.05). A positive correlation between vitamins E and C was detected at day 120 (r = 0.349, P < 0.01). Age and BCS did not affect the patterns of changes. Assuming that the ewes with P4 concentrations ≥2.5 ng/ml were pregnant, 95 out of 105 ewes (90.5%) were pregnant at day 120 of the study. Under the conditions of the present study with medium-to-low quality pastures as the main sources of feed, ewes of various ages and body conditions may suffer from oxidative stress during breeding and pregnancy.

Induction of a photomixotrophic plant cell culture of Helianthus annuus and optimization of culture conditions for improved α-tocopherol production.

PubMed

Geipel, Katja; Song, Xue; Socher, Maria Lisa; Kümmritz, Sibylle; Püschel, Joachim; Bley, Thomas; Ludwig-Müller, Jutta; Steingroewer, Juliane

2014-03-01

Tocopherols, collectively known as vitamin E, are lipophilic antioxidants, which are synthesized only by photosynthetic organisms. Due to their enormous potential to protect cells from oxidative damage, tocopherols are used, e.g., as nutraceuticals and additives in pharmaceuticals. The most biologically active form of vitamin E is α-tocopherol. Most tocopherols are currently produced via chemical synthesis. Nevertheless, this always results in a racemic mixture of different and less effective stereoisomers because the natural isomer has the highest biological activity. Therefore, tocopherols synthesized in natural sources are preferred for medical purposes. The annual sunflower (Helianthus annuus L.) is a well-known source for α-tocopherol. Within the presented work, sunflower callus and suspension cultures were established growing under photomixotrophic conditions to enhance α-tocopherol yield. The most efficient callus induction was achieved with sunflower stems cultivated on solid Murashige and Skoog medium supplemented with 30 g l(-1) sucrose, 0.5 mg l(-1) of the auxin 1-naphthalene acetic acid, and 0.5 mg l(-1) of the cytokinin 6-benzylaminopurine. Photomixotrophic sunflower suspension cultures were induced by transferring previously established callus into liquid medium. The effects of light intensity, sugar concentration, and culture age on growth rate and α-tocopherol synthesis rate were characterized. A considerable increase (max. 230%) of α-tocopherol production in the cells was obtained within the photomixotrophic cell culture compared to a heterotrophic cell culture. These results will be useful for improving α-tocopherol yields of plant in vitro cultures.

Hair-Growth-Promoting Effect of Conditioned Medium of High Integrin α6 and Low CD 71 (α6bri/CD71dim) Positive Keratinocyte Cells

PubMed Central

Won, Chong Hyun; Jeong, Yun-Mi; Kang, Sangjin; Koo, Tae-Sung; Park, So-Hyun; Park, Ki-Young; Sung, Young-Kwan; Sung, Jong-Hyuk

2015-01-01

Keratinocyte stem/progenitor cells (KSCs) reside in the bulge region of the hair follicles and may be involved in hair growth. Hair follicle dermal papilla cells (HFDPCs) and outer root sheath (ORS) cells were treated with conditioned medium (CM) of KSCs. Moreover, the effects of KSC-CM on hair growth were examined ex vivo and in vivo. A human growth factor chip array and RT-PCR were employed to identify enriched proteins in KSC-CM as compared with CM from keratinocytes. KSC-CM significantly increased the proliferation of HFDPCs and ORS cells, and increased the S-phase of the cell cycle in HFDPCs. KSC-CM led to the phosphorylation of ATK and ERK1/2 in both cell types. After subcutaneous injection of KSC-CM in C3H/HeN mice, a significant increase in hair growth and increased proliferation of hair matrix keratinocytes ex vivo was observed. We identified six proteins enriched in KSC-CM (amphiregulin, insulin-like growth factor binding protein-2, insulin-like growth factor binding protein-5, granulocyte macrophage-colony stimulating factor, Platelet-derived growth factor-AA, and vascular endothelial growth factor). A growth-factor cocktail that contains these six recombinant growth factors significantly increased the proliferation of HFDPCs and ORS cells and enhanced the hair growth of mouse models. These results collectively indicate that KSC-CM has the potential to increase hair growth via the proliferative capacity of HFDPCs and ORS cells. PMID:25706512

Bone culture research

NASA Technical Reports Server (NTRS)

Partridge, Nicola C.

1993-01-01

The experiments described are aimed at exploring PTH regulation of production of collagenase and protein inhibitors of collagenase (tissue inhibitors of metalloproteases, TIMP-1 and -2) by osteoblast-like osteosarcoma cells under conditions of weightlessness. The results of this work will contribute to information as to whether a microgravity environment alters the functions and responsiveness of the osteoblast. The objectives of the Bone Culture Research (BCR) experiment are: to observe the effects of microgravity on the morphology, rate of proliferation, and behavior of the osteoblastic cells, UMR 106-01; to determine whether microgravy affects the hormonal sensitivity of osteroblastic cells; and to measure the secretion of collagenase and its inhibitors into the medium under conditions of microgravity. The methods employed will consist of the following: the osteoblast-like cells, UMR-106-01, will be cultured in four NASDA cell culture chambers; two chambers will be subjected to microgravity on SL-J; two chambers will remain on the ground at KSC as ground controls but subjected to an identical set of culture conditions as on the shuttle; media will be changed four times; twice the cells will receive the hormone parathyroid hormone-related protein (PTHrP) and media collected; cells will be photographed under conditions of microgravity; and media and photographs will be analyzed upon return to determine whether functions of the cells changed.

40 CFR 52.719 - Identification of plan-Conditional approval.

Code of Federal Regulations, 2013 CFR

2013-07-01

... aerosol-based antiperspirants and deodorants. 35 IAC 233.205(a)(6)(A) erroneously provides two high-volatility VOC limits for aerosol-based antiperspirants when there should be both a high- and medium... limits for non aerosol-based antiperspirants when there should be both a high- and medium-volatility...

Quenching of a highly superheated porous medium by injection of water

NASA Astrophysics Data System (ADS)

Fichot, F.; Bachrata, A.; Repetto, G.; Fleurot, J.; Quintard, M.

2012-11-01

Understanding of two-phase flow through porous medium with intense phase change is of interest in many situations, including nuclear, chemical or geophysical applications. Intense boiling occurs when the liquid is injected into a highly superheated medium. Under such conditions, the heat flux extracted by the fluid from the porous medium is mainly governed by the nucleation of bubbles and by the evaporation of thin liquid films. Both configurations are possible, depending on local flow conditions and on the ratio of bubble size to pore size. The present study is motivated by the safety evaluation of light water nuclear reactors in case of a severe accident scenario, such as the one that happened in Fukushima Dai-ichi plant in March, 2011. If water sources are not available for a long period of time, the reactor core heats up due to the residual power and eventually becomes significantly damaged due to intense oxidation of metals and fragmentation of fuel rods resulting in the formation of a porous medium where the particles have a characteristic length-scale of 1 to 5 mm. The coolability of the porous medium will depend on the water flow rate which can enter the medium under the available driving head and on the geometrical features of the porous matrix (average pore size, porosity). Therefore, it is of high interest to evaluate the conditions for which the injection of water in such porous medium is likely to stop the progression of the accident. The present paper addresses the issue of modelling two-phase flow and heat transfers in a porous medium initially dry, where water is injected. The medium is initially at a temperature well above the saturation temperature of water. In a first part, a summary of existing knowledge is provided, showing the scarcity of models and experimental data. In a second part, new experimental results obtained in an IRSN facility are analysed. The experiment consists in a bed of steel particles that are heated up to 700°Cbefore injecting water. The facility is briefly described. The velocity of the "quench front" (location where particles are quickly cooled down) and the total pressure drop across the medium are estimated. The dependencies of those quantities with respect to the inlet water flow rate, the initial temperature of the medium and the diameter of particles are obtained. In a third part, a model is proposed, based on a previously developed model which is improved in order to take into account intense boiling regimes (in particular nucleate boiling). The model includes a function that takes into account the contact area between water and the particles which depends on the temperature of particles and on the void fraction. That function affects the local intensity of phase change. The model involves a few parameters which cannot be evaluated analytically. Those parameters are bounded, following the analysis of experimental data. Finally, the model is assessed by comparison of calculations with those new experimental data. The satisfactory agreement shows that the model is almost predictive in the range of parameters studied. The experimental results also show that the quench front becomes unstable under certain conditions. This is also analysed and compared with the predictions of the model.

Stirred tank bioreactor culture combined with serum-/xenogeneic-free culture medium enables an efficient expansion of umbilical cord-derived mesenchymal stem/stromal cells.

PubMed

Mizukami, Amanda; Fernandes-Platzgummer, Ana; Carmelo, Joana G; Swiech, Kamilla; Covas, Dimas T; Cabral, Joaquim M S; da Silva, Cláudia L

2016-08-01

Mesenchymal stem/stromal cells (MSC) are being widely explored as promising candidates for cell-based therapies. Among the different human MSC origins exploited, umbilical cord represents an attractive and readily available source of MSC that involves a non-invasive collection procedure. In order to achieve relevant cell numbers of human MSC for clinical applications, it is crucial to develop scalable culture systems that allow bioprocess control and monitoring, combined with the use of serum/xenogeneic (xeno)-free culture media. In the present study, we firstly established a spinner flask culture system combining gelatin-based Cultispher(®) S microcarriers and xeno-free culture medium for the expansion of umbilical cord matrix (UCM)-derived MSC. This system enabled the production of 2.4 (±1.1) x10(5) cells/mL (n = 4) after 5 days of culture, corresponding to a 5.3 (±1.6)-fold increase in cell number. The established protocol was then implemented in a stirred-tank bioreactor (800 mL working volume) (n = 3) yielding 115 million cells after 4 days. Upon expansion under stirred conditions, cells retained their differentiation ability and immunomodulatory potential. The development of a scalable microcarrier-based stirred culture system, using xeno-free culture medium that suits the intrinsic features of UCM-derived MSC represents an important step towards a GMP compliant large-scale production platform for these promising cell therapy candidates. Copyright © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Optimization of Culture Parameters for Maximum Polyhydroxybutyrate Production by Selected Bacterial Strains Isolated from Rhizospheric Soils.

PubMed

Lathwal, Priyanka; Nehra, Kiran; Singh, Manpreet; Jamdagni, Pragati; Rana, Jogender S

2015-01-01

The enormous applications of conventional non-biodegradable plastics have led towards their increased usage and accumulation in the environment. This has become one of the major causes of global environmental concern in the present century. Polyhydroxybutyrate (PHB), a biodegradable plastic is known to have properties similar to conventional plastics, thus exhibiting a potential for replacing conventional non-degradable plastics. In the present study, a total of 303 different bacterial isolates were obtained from soil samples collected from the rhizospheric area of three crops, viz., wheat, mustard and sugarcane. All the isolates were screened for PHB (Poly-3-hydroxy butyric acid) production using Sudan Black staining method, and 194 isolates were found to be PHB positive. Based upon the amount of PHB produced, the isolates were divided into three categories: high, medium and low producers. Representative isolates from each category were selected for biochemical characterization; and for optimization of various culture parameters (carbon source, nitrogen source, C/N ratio, different pH, temperature and incubation time periods) for maximizing PHB accumulation. The highest PHB yield was obtained when the culture medium was supplemented with glucose as the carbon source, ammonium sulphate at a concentration of 1.0 g/l as the nitrogen source, and by maintaining the C/N ratio of the medium as 20:1. The physical growth parameters which supported maximum PHB accumulation included a pH of 7.0, and an incubation temperature of 30 degrees C for a period of 48 h. A few isolates exhibited high PHB accumulation under optimized conditions, thus showing a potential for their industrial exploitation.

In vitro cultivation of Hysterothylacium aduncum (Nematoda: Anisakidae) from 3rd-stage larvae to egg-laying adults.

PubMed

Iglesias, L; Valero, A; Gálvez, L; Benítez, R; Adroher, F J

2002-11-01

This is the first demonstration of the in vitro development of the 3rd-stage larvae (L3) of Hysterothylacium aduncum to the adult. This was achieved in a semi-defined medium that is easy to prepare and to reproduce. The L3, collected from the peritoneal cavity of horse mackerel (Trachurus trachurus), were individually inoculated into RPMI-1640 medium +20% heat-inactivated fetal bovine serum (IFBS). It has been demonstrated that the optimum temperature for development is around 13 degrees C and is stimulated by the presence of 5% CO2 in the growth atmosphere, increasing the percentage moulting to the 4th larval stage (L4) by 1.9-fold (from 44 to 82%) and the average survival of the nematodes by 1.6 times (from 60 to 96 days). When the larvae were grown at different pHs, optimum development occurred at pH 4.0. Under these conditions, all the larvae moulted to the L4 and more than two-thirds transformed to the adult stage--in which 25-30% of the females laid eggs--and reached an average survival of over 4 months. When this medium was supplemented with 1% (w/v) of commercial pepsin, all the larvae reached the adult stage, at least 45% of the females oviposited, laying around 12-fold more eggs per female than in the medium without pepsin. The mean size of the eggs (non-fertilized) obtained was 56.8 x 47.6 microm. The mean length of the adult males obtained was between 3.2 and 5.2 cm and the females were between 3.0 and 6.5 cm. The adult specimens were morphologically identified as Hysterothylacium aducum aduncum. This culture medium (RPMI-1640+20% (v/v) IFBS+1 commercial pepsin, at pH 4.0, 13 degrees C and 5% CO2 in air) could facilitate the identification of at least some of the larvae of the genus Hysterothylacium--and perhaps other anisakids--for which the specific identification and the biological study of these parasites is often difficult.

A comparison of legionella and other bacteria concentrations in cooling tower water

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

Cappabianca, R.M.; Jurinski, N.B.; Jurinski, J.B.

1994-05-01

A field study was conducted in which water samples collected from air conditioning cooling water reservoirs of high-rise buildings throughout an urban area were assayed for Legionella and for total bacteria. Buildings included within the study had ongoing biocidal treatment programs for the cooling towers. Separate sample analyses were performed to measure the viable colony concentrations of total bacteria and of Legionella in the process waters. The occurrence and viable counts of Legionella in 304 environmental water samples were determined by inoculating them onto plates of buffered charcoal yeast extract (BCYE) agar medium (a presumptive screening method). The samples weremore » collected during summer months between July and September. BCYE plate cultures of 50 (16.4%) of the samples yielded Legionella with viable counts ranging from 2 to 608 colony forming units per milliliter. In the water samples, 281 (92.4%) yielded viable counts of bacteria that ranged from 9 to 1.2 x 10{sup 6} per milliliter. This study demonstrates that Legionella are commonly present in the water of air conditioning cooling towers and that there is no significant correlation between concurrently sampled culture plate counts of Legionella and total bacteria plate counts. Correspondingly, there is no demonstrated validity for use of total bacterial counts as an inferential surrogate for the concentration of Legionella in the water. 19 refs., 3 figs., 1 tab.« less

DEM simulation of dendritic grain random packing: application to metal alloy solidification

NASA Astrophysics Data System (ADS)

Olmedilla, Antonio; Založnik, Miha; Combeau, Hervé

2017-06-01

The random packing of equiaxed dendritic grains in metal-alloy solidification is numerically simulated and validated via an experimental model. This phenomenon is characterized by a driving force which is induced by the solid-liquid density difference. Thereby, the solid dendritic grains, nucleated in the melt, sediment and pack with a relatively low inertia-to-dissipation ratio, which is the so-called Stokes number. The characteristics of the particle packed porous structure such as solid packing fraction affect the final solidified product. A multi-sphere clumping Discrete Element Method (DEM) approach is employed to predict the solid packing fraction as function of the grain geometry under the solidification conditions. Five different monodisperse noncohesive frictionless particle collections are numerically packed by means of a vertical acceleration: a) three dendritic morphologies; b) spheres and c) one ellipsoidal geometry. In order to validate our numerical results with solidification conditions, the sedimentation and packing of two monodisperse collections (spherical and dendritic) is experimentally carried out in a viscous quiescent medium. The hydrodynamic similarity is respected between the actual phenomenon and the experimental model, that is a low Stokes number, o(10-3). In this way, the experimental average solid packing fraction is employed to validate the numerical model. Eventually, the average packing fraction is found to highly depend on the equiaxed dendritic grain sphericity, with looser packings for lower sphericity.

Changes in DnaA-dependent gene expression contribute to the transcriptional and developmental response of Bacillus subtilis to manganese limitation in Luria-Bertani medium.

PubMed

Hoover, Sharon E; Xu, Weihong; Xiao, Wenzhong; Burkholder, William F

2010-08-01

The SOS response to DNA damage in bacteria is a well-known component of the complex transcriptional responses to genotoxic environmental stresses such as exposure to reactive oxygen species, alkylating agents, and many of the antibiotics targeting DNA replication. However, bacteria such as Bacillus subtilis also respond to conditions that perturb DNA replication via a transcriptional response mediated by the replication initiation protein DnaA. In addition to regulating the initiation of DNA replication, DnaA directly regulates the transcription of specific genes. Conditions that perturb DNA replication can trigger the accumulation of active DnaA, activating or repressing the transcription of genes in the DnaA regulon. We report here that simply growing B. subtilis in LB medium altered DnaA-dependent gene expression in a manner consistent with the accumulation of active DnaA and that this was part of a general transcriptional response to manganese limitation. The SOS response to DNA damage was not induced under these conditions. One of the genes positively regulated by DnaA in Bacillus subtilis encodes a protein that inhibits the initiation of sporulation, Sda. Sda expression was induced as cells entered stationary phase in LB medium but not in LB medium supplemented with manganese, and the induction of Sda inhibited sporulation-specific gene expression and the onset of spore morphogenesis. In the absence of Sda, manganese-limited cells initiated spore development but failed to form mature spores. These data highlight that DnaA-dependent gene expression may influence the response of bacteria to a range of environmental conditions, including conditions that are not obviously associated with genotoxic stress.

Convection induced by radiative cooling of a layer of participating medium

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

Prasanna, Swaminathan, E-mail: prasannaswam@gmail.com; Venkateshan, S. P., E-mail: spv@iitm.ac.in

2014-05-15

Simulations and experiments have been conducted to study the effect of radiative cooling on natural convection in a horizontal layer of a participating medium enclosed between isothermal opaque wall and radiatively transparent wall and exposed to a cold background. The study is of relevance to a nocturnal boundary layer under clear and calm conditions. The focus of the study is to capture the onset of convection caused by radiative cooling. The experiments have been designed to mimic the atmospheric radiative boundary conditions, and hence decoupling convection and radiation boundary conditions. Planck number Pl and optical thickness of the layer τ{submore » H} are the two important parameters that govern the interaction between radiation and convection. The radiation-convection coupling is a strong function of length scale. Convection sets up within first few seconds for all the experiments. Strong plume like convection is observed for the experimental conditions used in the present study. Both simulations and experiments confirm that radiative cooling increases substantially with decrease in emissivity of the bottom wall. Radiative cooling is strongly influenced by the nongray nature of the participating medium, especially when strong emission from the medium escapes to space, in the window region of the atmosphere. Accurate representation of radiative properties is critical. Linear stability analysis of onset of convection indicates that radiation stabilizes convection as Pl decreases. The observations are similar to the case of Rayleigh Bénard convection in a radiating gas. However, for both experimental and numerical conditions, the observed Rayleigh numbers are much greater than the critical Rayleigh number. To conclude, the role of radiation is to drive and sustain convection in the unstable layer.« less

[Mass-transfer, utilization, and diffusion of oxygen in skeletal muscles of the stenohaline goby Gobius cobitus Pallas under conditions of hypoosmotic medium].

PubMed

Soldatov, A A

2012-01-01

Effect of hypoosmotic conditions of medium on oxygen regime of skeletal muscles of the stenohalin goby Gobius cobitus Pallas was studied under conditions of experiment. The control fish group was maintained at 12-14 %o, the experimental one - at 4.8-5.6 per thousand. Duration of the experiment - 44-45 days, water temperature - 15 +/- 1 degrees C, photoperiod - 12 day/12 night. It was established that under conditions of external hypoosmia there occurred hydration of the goby skeletal muscles and a decrease of their diffusion capability with respect to oxygen. The latter was accompanied by the tissue P(O2) decrease, which is indicated by low values of P(O2) in the venous blood outflowing from muscles. For the first 14-16 days of adaptation to the hypoosmotic medium there were restricted processes of mass transfer and oxygen utilization, which was associated with a decrease of the voluminous tissue blood flow and the blood oxygen concentration. These changes occurred on the background of the blood plasma hydration and a decrease of the number of circulated erythrocytes, and then they were completely compensated.

Bank Size and Small- and Medium-sized Enterprise (SME) Lending: Evidence from China

PubMed Central

SHEN, YAN; SHEN, MINGGAO; XU, ZHONG; BAI, YING

2014-01-01

Summary Using panel data collected in 2005, we evaluate how bank size, discretion over credit, incentive schemes, competition, and the institutional environment affect lending to small- and medium-sized enterprises in China. We deal with the endogeneity problem using instrumental variables, and a reduced-form approach is also applied to allow for weak instruments in estimation. We find that total bank asset is an insignificant factor for banks’ decision on small- and medium-enterprise (SME) lending, but more local lending authority, more competition, carefully designed incentive schemes, and stronger law enforcement encourage commercial banks to lend to SMEs. PMID:26052179

Tracing Thermal Creep Through Granular Media

NASA Astrophysics Data System (ADS)

Steinpilz, Tobias; Teiser, Jens; Koester, Marc; Schywek, Mathias; Wurm, Gerhard

2017-08-01

A temperature gradient within a granular medium at low ambient pressure drives a gas flow through the medium by thermal creep. We measured the resulting air flow for a sample of glass beads with particle diameters between 290 μ m and 420 μ m for random close packing. Ambient pressure was varied between 1 Pa and 1000 Pa. The gas flow was quantified by means of tracer particles during parabolic flights. The flow varies systematically with pressure between 0.2 cm/s and 6 cm/s. The measured flow velocities are in quantitative agreement to model calculations that treat the granular medium as a collection of linear capillaries.

Monitoring of bisphenols in canned tuna from Italian markets.

PubMed

Fattore, Margherita; Russo, Giacomo; Barbato, Francesco; Grumetto, Lucia; Albrizio, Stefania

2015-09-01

Monitoring of food contamination from bisphenols is a necessary process for the consumers' risk assessment. A method for the quali-quantitative analysis of Bisphenol A (BPA), Bisphenol B (BPB), Bisphenol A Diglycidyl Ether (BADGE), and Bisphenol F Diglycidyl Ether (BFDGE), by liquid chromatography with fluorescence detection (LC-FD), was performed and validated for their determination in 33 samples of tuna fish, canned in either oil or aqueous medium. Samples were collected in Italian markets. Tuna and the correspondent preservation medium were analyzed separately. Detected levels of bisphenols ranged from 19.1 to 187.0 ng/g in tuna matrix and from 6.3 to 66.9 ng/mL in oil medium. No bisphenols were found in aqueous medium. At least one of the analytes was found in 83% of the tuna samples in oil medium, whereas tuna samples in aqueous medium showed BPA alone in 67% of samples. 21% of the oil medium samples resulted positive for at least one bisphenol. On the basis of measured concentrations and general daily ingestion rate of canned tuna fish, the probable daily intake of BPA for Italian population was calculated. Copyright © 2015 Elsevier Ltd. All rights reserved.

In vitro propagation of Stevia rebaudina plants using multiple shoot culture.

PubMed

Nepovím, A; Vanek, T

1998-12-01

A multiple shoot culture was induced from nodal segments on MS medium containing half concentration of macroelements, 1% sucrose, and supplemented with NAA (0.01 mg/l). A bioreactor with hormone-free MS medium (300 ml) was inoculated with 1.5 g of the multiple shoot culture and cultivated for a month. The cultivating process of the multiple shoot culture in the bioreactor and the transfer into ex vitro conditions took about 8-9 weeks and produced approx. 600 new seedlings, that could be transferred from greenhouse to field conditions.

August gamma Cepheids (523-AGC)

NASA Astrophysics Data System (ADS)

Roggemans, Paul

2018-02-01

Favorable weather conditions between 19 August and 5 September 2017 enabled the CAMS BeNeLux network to collect 3189 orbits. A radiant concentration was spotted which was identified as the August gamma Cepheids (523-AGC). An independent search on a selection from all available meteor orbit lists coming from the suspect radiant area and velocity range was made. This resulted in 283 similar orbits, radiating from R.A. 358.4° and Decl. +76.2° with a geocentric velocity of 43.7 km/s in a time lapse between 146° and 165° in solar longitude with best activity at 155.7°. The orbital elements match perfectly with previously published results. There is no indication for any periodicity in the shower displays from year to year. The AGC-meteors are remarkably rich in bright meteors and rather deficient in faint meteors. Being detected independently from orbital data collected by different video networks, confirmed by 283 orbits with a medium threshold D criterion DD < 0.08 and 125 orbits with a high threshold of DD < 0.04, this minor shower could be considered to be listed as an established meteor shower.

Growth and differentiation of a murine interleukin-3-producing myelomonocytic leukemia cell line in a protein-free chemically defined medium.

PubMed

Kajigaya, Y; Ikuta, K; Sasaki, H; Matsuyama, S

1990-10-01

We established the continuous growth of WEHI-3B D+ cells in protein-free chemically defined F-12 medium by stepwise decreases in the concentration of fetal calf serum. This cell line, designated as WEHI-3B-Y1, has now been propagated in protein-free F-12 medium for 3 years. The population-doubling time of the cells in culture is about 24 hr. WEHI-3B-Y1 cells are immature undifferentiated cells which show positive staining for naphthol ASD chloroacetate esterase and alpha-naphthyl butyrate esterase and spontaneously exhibit a low level of differentiation to mature granulocytes and macrophages. Medium conditioned by WEHI-3B-Y1 cells stimulated the proliferation of an interleukin-3 (IL-3)-dependent FDCP-2 cell line. This conditioned medium was shown to have erythroid burst-promoting activity when assayed using normal murine bone marrow. The colony formation of WEHI-3B-Y1 cells in semi-solid agar culture was not stimulated by purified recombinant human granulocyte colony-stimulating factor (rhG-CSF). However, in the presence of human transferrin, rhG-CSF enhanced the number of colonies of WEHI-3B-Y1 cells but did not induce their differentiation. These results suggest that WEHI-3B-Y1 cells cultured in protein-free medium produced murine IL-3. In addition, human G-CSF enhanced the clonal growth but did not induce the differentiation of WEHI-3B-Y1 cells cultured in serum-free medium.

Economic Learning Media Development Based on Local Locality

ERIC Educational Resources Information Center

Hadi, Rizali; Supriyanto; Hasanah, Mahmudah

2017-01-01

This study aims to describe the learning medium of economic education at senior High School in Banjarmasin with media based on local wisdom. This research uses qualitative method as developed by Miles & Huberman, starting from data collection, data reduction data display, and then made conclusion. Data were collected in the order of Basic…

78 FR 48413 - Proposed Information Collection; Comment Request; Voluntary Self-Disclosure of Antiboycott...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-08-08

... Respondents: 15. Estimated Time per Response: 10 for medium-size companies; 600 hours for large-size companies... proper performance of the functions of the agency, including whether the information shall have practical... proposed collection of information; (c) ways to enhance the quality, utility, and clarity of the...

Preparing the Production of a New Product in Small and Medium-Sized Enterprises by Using the Method of Projects Management

NASA Astrophysics Data System (ADS)

Bijańska, Jolanta; Wodarski, Krzysztof; Wójcik, Janusz

2016-06-01

Efficient and effective preparation the production of new products is important requirement for a functioning and development of small and medium-sized enterprises. One of the methods, which support the fulfilment of this condition is project management. This publication presents the results of considerations, which are aimed at developing a project management model of preparation the production of a new product, adopted to specificity of small and medium-sized enterprises.

Platelet concentrates for transfusion-metabolic and storage aspects.

PubMed

Farrugia, A

1994-01-01

Transfusion of platelets concentrated from donated blood is an established therapeutic modality in clinical medicine. Over the past 25 years much effort has gone into optimising the conditions for the collection, preparation and storage of platelets for transfusion. Despite significant advances, platelet production is still a costly process requiring a dedicated environment and the use of specially formulated plastic storage containers. A progressive lesion over storage limits the shelf life and the availability of donated platelets, while the need to store platelets in the donor's autologous plasma also results in a loss of valuable fresh plasma for fractionation. Recent studies have addressed the issues of platelet quality and plasma economy by examining the possibility of storing platelets in a synthetic medium. Platelets stored in a variety of crystalloid solutions have been shown to retain in vitro and in vivo properties equivalent or superior to platelets stored in autologous donor plasma. Some additional insight has been gained on the metabolic patterns of stored platelets. In particular, studies have shown that, under these conditions, platelets are unable to oxidise dextrose to any significant extent, and that dextrose is invariably broken down to lactate, irrespective of the oxygen tensions in the platelet's environment. This in turn leads to the metabolic lesion of platelet storage, whereby low pH results in loss of platelet viability. Platelets stored in synthetic dextrose-free media are capable of maintaining aerobic ATP generation, and acetate-a component of many media studied-has been shown to be metabolised by platelets. Similarly, platelets prepared from blood collected into a dextrose-free anticoagulant have satisfactory properties both when suspended in autologous plasma or in a dextrose-free synthetic medium. The requirements for storage in special, high gas-permeable, containers, and for constant agitation during storage, were both found to be unnecessary when dextrose was excluded from the platelet's environment. These developments suggest that manipulation of the platelet's metabolic pattern during blood bank storage may allow significant benefits in plasma economy as well as in decreasing the cost of platelet delivery to patients.

Application of a paper based device containing a new culture medium to detect Vibrio cholerae in water samples collected in Haiti.

PubMed

Briquaire, Romain; Colwell, Rita R; Boncy, Jacques; Rossignol, Emmanuel; Dardy, Aline; Pandini, Isabelle; Villeval, François; Machuron, Jean-Louis; Huq, Anwar; Rashed, Shah; Vandevelde, Thierry; Rozand, Christine

2017-02-01

Cholera is now considered to be endemic in Haiti, often with increased incidence during rainy seasons. The challenge of cholera surveillance is exacerbated by the cost of sample collection and laboratory analysis. A diagnostic tool is needed that is low cost, easy-to-use, and able to detect and quantify Vibrio cholerae accurately in water samples within 18-24h, and perform reliably in remote settings lacking laboratory infrastructure and skilled staff. The two main objectives of this study were to develop and evaluate a new culture medium embedded in a new diagnostic tool (PAD for paper based analytical device) for detecting Vibrio cholerae from water samples collected in Haiti. The intent is to provide guidance for corrective action, such as chlorination, for water positive for V. cholerae epidemic strains. For detecting Vibrio cholerae, a new chromogenic medium was designed and evaluated as an alternative to thiosulfate citrate bile salts sucrose (TCBS) agar for testing raw water samples. Sensitivity and specificity of the medium were assessed using both raw and spiked water samples. The Vibrio cholerae chromogenic medium was proved to be highly selective against most of the cultivable bacteria in the water samples, without loss of sensitivity in detection of V. cholerae. Thus, reliability of this new culture medium for detection of V. cholerae in the presence of other Vibrio species in water samples offers a significant advantage. A new paper based device containing the new chromogenic medium previously evaluated was compared with reference methods for detecting V. cholerae from spiked water sample. The microbiological PAD specifications were evaluated in Haiti. More precisely, a total of 185 water samples were collected at five sites in Haiti, June 2014 and again in June 2015. With this new tool, three V. cholerae O1 and 17 V. cholerae non-O1/O139 strains were isolated. The presence of virulence-associated and regulatory genes, including ctxA, zot, ace, and toxR, was confirmed using multiplex PCR. The three V. cholerae O1 isolates were positive for three of the four virulence-associated and regulatory genes. Twelve of the V. cholerae non-O1/O139 isolates were found to carry toxR, but none were ctxA+, zot+, or ace+. However, six of the V. cholerae non-O1/O139 isolates were resistant to penicillin, ampicillin, trimethoprim/sulfamethoxazole, nalidixic acid, and ciprofloxacin. The paper based analytical device (PAD) provides advantages in that standard culture methods employing agar plates are not required. Also, intermediary isolation steps were not required, including transfer to selective growth media, hence these steps being omitted reduced time to results. Furthermore, experienced technical skills also were not required. Thus, PAD is well suited for resource-limited settings. Copyright © 2016 Elsevier B.V. All rights reserved.

Metamaterial-based lossy anisotropic epsilon-near-zero medium for energy collimation

NASA Astrophysics Data System (ADS)

Shen, Nian-Hai; Zhang, Peng; Koschny, Thomas; Soukoulis, Costas M.

2016-06-01

A lossy anisotropic epsilon-near-zero (ENZ) medium may lead to a counterintuitive phenomenon of omnidirectional bending-to-normal refraction [S. Feng, Phys. Rev. Lett. 108, 193904 (2012), 10.1103/PhysRevLett.108.193904], which offers a fabulous strategy for energy collimation and energy harvesting. Here, in the scope of effective medium theory, we systematically investigate two simple metamaterial configurations, i.e., metal-dielectric-layered structures and the wire medium, to explore the possibility of fulfilling the conditions of such an anisotropic lossy ENZ medium by playing with materials' parameters. Both realistic metamaterial structures and their effective medium equivalences have been numerically simulated, and the results are in excellent agreement with each other. Our study provides clear guidance and therefore paves the way towards the search for proper designs of anisotropic metamaterials for a decent effect of energy collimation and wave-front manipulation.

Dielectric recovery mechanism of pressurized carbon dioxide at liquid and supercritical phases

NASA Astrophysics Data System (ADS)

Tanoue, Hiroyuki; Furusato, Tomohiro; Imamichi, Takahiro; Ota, Miyuki; Katsuki, Sunao; Akiyama, Hidenori

2015-09-01

Estimates of dielectric recovery rates of supercritical (SC) and liquid carbon dioxide (CO2) were derived with focus on highly-repetitive pulsed power switching mediums. Calculated results suggest that recovery time of SC and liquid CO2 are approximately 50 times shorter than that of water and oils. Prior to 10 µs after breakdown, recovery rates in neither SC nor liquid CO2 reached 100%, though the recovery rate in SC CO2 was higher than that of liquid CO2. To examine causes of recovery rate differences, each dielectric recovery process in SC and liquid CO2 was observed by laser shadowgraph technique. These shadowgraph images suggest two factors explaining dielectric recovery rate differences between these medium conditions: 1) thermodynamic property differences between medium conditions, and 2) differences in the low density region recovery mechanism.

"Ant" and "grasshopper" life-history strategies in Saccharomyces cerevisiae.

PubMed

Spor, Aymé; Wang, Shaoxiao; Dillmann, Christine; de Vienne, Dominique; Sicard, Delphine

2008-02-13

From the evolutionary and ecological points of view, it is essential to distinguish between the genetic and environmental components of the variability of life-history traits and of their trade-offs. Among the factors affecting this variability, the resource uptake rate deserves particular attention, because it depends on both the environment and the genetic background of the individuals. In order to unravel the bases of the life-history strategies in yeast, we grew a collection of twelve strains of Saccharomyces cerevisiae from different industrial and geographical origins in three culture media differing for their glucose content. Using a population dynamics model to fit the change of population size over time, we estimated the intrinsic growth rate (r), the carrying capacity (K), the mean cell size and the glucose consumption rate per cell. The life-history traits, as well as the glucose consumption rate, displayed large genetic and plastic variability and genetic-by-environment interactions. Within each medium, growth rate and carrying capacity were not correlated, but a marked trade-off between these traits was observed over the media, with high K and low r in the glucose rich medium and low K and high r in the other media. The cell size was tightly negatively correlated to carrying capacity in all conditions. The resource consumption rate appeared to be a clear-cut determinant of both the carrying capacity and the cell size in all media, since it accounted for 37% to 84% of the variation of those traits. In a given medium, the strains that consume glucose at high rate have large cell size and low carrying capacity, while the strains that consume glucose at low rate have small cell size but high carrying capacity. These two contrasted behaviors may be metaphorically defined as "ant" and "grasshopper" strategies of resource utilization. Interestingly, a strain may be "ant" in one medium and "grasshopper" in another. These life-history strategies are discussed with regards to yeast physiology, and in an evolutionary perspective.

Medium Truck Duty Cycle Data from Real-World Driving Environments: Final Report

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

Lascurain, Mary Beth; Franzese, Oscar; Capps, Gary J

2012-11-01

Since the early part of the 20th century, the US trucking industry has provided a safe and economical means of moving commodities across the country. At present, nearly 80% of US domestic freight movement involves the use of trucks. The US Department of Energy (DOE) is spearheading a number of research efforts to improve heavy vehicle fuel efficiencies. This includes research in engine technologies (including hybrid and fuel cell technologies), lightweight materials, advanced fuels, and parasitic loss reductions. In addition, DOE is developing advanced tools and models to support heavy vehicle research and is leading the 21st Century Truck Partnershipmore » and the SuperTruck development effort. Both of these efforts have the common goal of decreasing the fuel consumption of heavy vehicles. In the case of SuperTruck, a goal of improving the overall freight efficiency of a combination tractor-trailer has been established. This Medium Truck Duty Cycle (MTDC) project is a critical element in DOE s vision for improved heavy vehicle energy efficiency; it is unique in that there is no other existing national database of characteristic duty cycles for medium trucks based on collecting data from Class 6 and 7 vehicles. It involves the collection of real-world data on medium trucks for various situational characteristics (e.g., rural/urban, freeway/arterial, congested/free-flowing, good/bad weather) and looks at the unique nature of medium trucks drive cycles (stop-and-go delivery, power takeoff, idle time, short-radius trips). This research provides a rich source of data that can contribute to the development of new tools for FE and modeling, provide DOE a sound basis upon which to make technology investment decisions, and provide a national archive of real-world-based medium-truck operational data to support energy efficiency research. The MTDC project involved a two-part field operational test (FOT). For the Part-1 FOT, three vehicles each from two vocations (urban transit and dry-box delivery) were instrumented for the collection of one year of operational data. The Part-2 FOT involved the towing and recovery and utility vocations for a second year of data collection. The vehicles that participated in the MTDC project did so through gratis partnerships in return for early access to the results of this study. Partnerships such as these are critical to FOTs in which real-world data is being collected. In Part 1 of the project, Oak Ridge National Laboratory (ORNL) established partnerships with the H.T. Hackney Company (HTH), one of the largest wholesale distributors in the country, distributing products to 21 states; and with Knoxville Area Transit (KAT), the city of Knoxville s transit system, which operates across Knoxville and parts of Knox County. These partnerships and agreements provided ORNL access to three Class-7 day-cab tractors that regularly haul 28 ft pup trailers (HTH) and three Class-7 buses for the collection of duty cycle data. In addition, ORNL collaborated with the Federal Motor Carrier Safety Administration (FMCSA) to determine if there were possible synergies between this duty cycle data collection effort and FMCSA s need to learn more about the operation and duty cycles of medium trucks. FMCSA s primary interest was in collecting safety data relative to the driver, carrier, and vehicle. In Part 2 of the project, ORNL partnered with the Knoxville Utilities Board, which made available three Class-8 trucks. Fountain City Wrecker Service was also a Part 2 partner, providing three Class-6 rollback trucks. In order to collect the duty cycle and safety-related data, ORNL developed a data acquisition system (DAS) that was placed on each test vehicle. Each signal recorded in this FOT was collected by means of one of the instruments incorporated into each DAS. Other signals were obtained directly from the vehicle s J1939 and J1708 data buses. A VBOX II Lite collected information available from a global positioning system (GPS), including speed, acceleration, and spatial location information at a rate of 5 Hz for the Part 1 FOT. For the Part 2 FOT, this information was obtained from DAS-based GPS instrumentation. The Air-Weigh LoadMaxx, a self-weighing system that determines the vehicle s gross weight by means of pressure transducers, was used to collect vehicle payload information for the combination, urban transit, and towing and recovery vehicles. A cellular modem, the Raven X EVDO V4221, facilitated the communication between the eDAQ-lite (the data collection engine of the system) and the user. The modem functioned as a wireless gateway, allowing data retrievals and system checks to be performed remotely. Also, in partnership with FMCSA, two additional safety sensors were installed on the combination vehicles: the MGM e-Stroke brake monitoring system and the Tire SafeGuard tire pressure monitoring system. All of these sensors posted data to the J1939 data bus, enabling the signals to be read withou...« less

Creatine enhances the duration of sperm capacitation: a novel factor for improving in vitro fertilization with small numbers of sperm.

PubMed

Umehara, Takashi; Kawai, Tomoko; Goto, Masaaki; Richards, JoAnne S; Shimada, Masayuki

2018-06-01

Why are many sperm required for successful fertilization of oocytes in vitro, even though fertilization occurs in vivo when only a few sperm reach the oocyte? Creatine produced in the ovary promotes efficient fertilization in vivo; however, in vitro, creatine is not contained in the in vitro fertilization (IVF) medium. The IVF medium enables capacitation of sperm. However, the IVF medium does not fully mimic the in vivo environment during fertilization. Consequently, fertilization in vitro is more inefficient than in the oviduct. Follicular and oviductal fluids were collected and then analyzed for creatine and glucose levels. To determine the physiological functions of creatine, the creatine antagonist 3-guanidinopropionic acid (GPA) was injected into hormonally primed mice. Using conventional IVF protocols, sperm were pre-incubated in IVF medium with creatine and then co-cultured with 10 ovulated cumulus-oocyte complexes (1-1000 per oocyte) in 50 μl medium droplets. Glucose and creatine levels were measured using commercial enzymatic assay kits. The effect of creatine in vivo was assessed by mating experiments using mice treated with or without GPA just before ovulation. To assess the functions of sperm incubated in IVF medium containing creatine, we analyzed (1) the motility of sperm using computer-assisted sperm assay, (2) the capacitation level of sperm by western blot analyses, and (3) the condition of sperm acrosomes by peanut agglutinin lectin-FITC staining. Oviductal creatine levels were significantly increased following ovulation. Injecting mice with GPA just before ovulation significantly reduced the number of fertilized oocytes. The addition of creatine to IVF medium enhanced sperm capacitation by increasing ATP levels. Successful fertilization was achieved with as few as five sperm/oocyte in the creatine group, and the number of fertilized oocytes was significantly higher than in the control without creatine (P < 0.01). In the present study, a pharmacological approach, creatine antagonist (GPA) treatment, but not a knockout mouse model, was used to understand the role of creatine in vivo. The role of creatine in fertilization processes can only be shown in a mouse model. A modified IVF technique using creatine-containing medium was developed and shown to markedly improve fertilization with small numbers of sperm. This approach has the potential to be highly beneficial for human assisted reproductive technologies, especially for patients with a limited number of good quality sperm. This work was supported in part by JSPS KAKENHI Grant numbers JP24688028, JP16H05017 (to M.S.), and JP15J05331 (to T.U.), the Japan Agency for Medical Research and Development (AMED) (16gk0110015h0001 to M.S.), and National Institutes of Health (NIH-HD-076980 to J.S.R). The authors have nothing to disclose.

Degradation testing of Mg alloys in Dulbecco's modified eagle medium: Influence of medium sterilization.

PubMed

Marco, Iñigo; Feyerabend, Frank; Willumeit-Römer, Regine; Van der Biest, Omer

2016-05-01

This work studies the in vitro degradation of Mg alloys for bioabsorbable implant applications under near physiological conditions. For this purpose, the degradation behaviour of Mg alloys in Dulbecco's modified eagle medium (DMEM) which is a commonly used cell culture medium is analysed. Unfortunately, DMEM can be contaminated by microorganisms, acidifying the medium and accelerating the Mg degradation process by dissolution of protective degradation layers, such as (Mgx,Cay)(PO4)z. In this paper the influence of sterilization by applying UV-C radiation and antibiotics (penicillin/streptomycin) is analysed with two implant material candidates: Mg-Gd and Mg-Ag alloys; and pure magnesium as well as Mg-4Y-3RE as a reference. Copyright © 2016 Elsevier B.V. All rights reserved.

Transmission versus reflectance spectroscopy for quantitation.

PubMed

Gardner, Craig M

2018-01-01

The objective of this work was to compare the accuracy of analyte concentration estimation when using transmission versus diffuse reflectance spectroscopy of a scattering medium. Monte Carlo ray tracing of light through the medium was used in conjunction with pure component absorption spectra and Beer-Lambert absorption along each ray's pathlength to generate matched sets of pseudoabsorbance spectra, containing water and six analytes present in skin. PLS regression models revealed an improvement in accuracy when using transmission compared to reflectance for a range of medium thicknesses and instrument noise levels. An analytical expression revealed the source of the accuracy degradation with reflectance was due both to the reduced collection efficiency for a fixed instrument etendue and to the broad pathlength distribution that detected light travels in the medium before exiting from the incident side. (2018) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE).

Biodiversity analysis by polyphasic study of marine bacteria associated with biocorrosion phenomena.

PubMed

Boudaud, N; Coton, M; Coton, E; Pineau, S; Travert, J; Amiel, C

2010-07-01

A polyphasic approach was used to study the biodiversity bacteria associated with biocorrosion processes, in particular sulfate-reducing bacteria (SRB) and thiosulfate-reducing bacteria (TRB) which are described to be particularly aggressive towards metallic materials, notably via hydrogen sulfide release. To study this particular flora, an infrared spectra library of 22 SRB and TRB collection strains were created using a Common Minimum Medium (CMM) developed during this study and standardized culture conditions. The CMM proved its ability to allow for growth of both SRB and TRB strains. These sulfurogen collection strains were clearly discriminated and differentiated at the genus level by fourier transform infrared (FT-IR) spectroscopy. In a second step, infrared spectra of isolates, recovered from biofilms formed on carbon steel coupons immersed for 1 year in three different French harbour areas, were compared to the infrared reference spectra library. In parallel, molecular methods (M13-PCR and 16S rRNA gene sequencing) were used to qualitatively evaluate the intra- and inter-species genetic diversity of biofilm isolates. The biodiversity study indicated that strains belonging to the Vibrio genus were the dominant population; strains belonging to the Desulfovibrio genus (SRB) and Peptostreptococcaceae were also identified. Overall, the combination of the FT-IR spectroscopy and molecular approaches allowed for the taxonomic and ecological study of a bacterial flora, cultivated on CMM, associated with microbiology-induced corrosion (MIC) processes. Via the use of the CMM medium, the culture of marine bacteria (including both SRB and TRB bacteria) was allowed, and the implication of nonsulforogen bacteria in MIC was observed. Their involvement in the biocorrosion phenomena will have to be studied and taken into account in the future. © 2009 The Authors. Journal compilation © 2009 The Society for Applied Microbiology.

Advanced technique for long term culture of epithelia in a continuous luminal-basal medium gradient.

PubMed

Schumacher, Karl; Strehl, Raimund; de, Vries Uwe; Minuth, Will W

2002-02-01

The majority of epithelia in our organism perform barrier functions on being exposed to different fluids at the luminal and basal sides. To simulate this natural situation under in vitro conditions for biomaterial testing and tissue engineering the epithelia have to withstand mechanical and fluid stress over a prolonged period of time. Leakage, edge damage and pressure differences in the culture system have to be avoided so that the epithelial barrier function is maintained. Besides, the environmental influences on important cell biological features such as, sealing or transport functions, have to remain upregulated and a loss of characteristics by dedifferentiation is prevented. Our aim is to expose embryonic renal collecting duct (CD) epithelia as model tissue for 14 days to fluid gradients and to monitor the development of tissue-specific features. For these experiments, cultured embryonic epithelia are placed in tissue carriers and in gradient containers, where different media are superfused at the luminal and basal sides. Epithelia growing on the tissue carriers act as a physiological barrier during the whole culture period. To avoid mechanical damage of the tissue and to suppress fluid pressure differences between the luminal and basal compartments improved transport of the medium and an elimination of unilaterally accumulated gas bubbles in the gradient container compartments by newly developed gas expander modules is introduced. By the application of these tools the yield of embryonic renal collecting duct epithelia with intact barrier function on a fragile natural support material could be increased significantly as compared to earlier experiments. Epithelia treated with a luminal NaCl load ranging from 3 to 24 mmol l were analyzed by immunohistochemical methods to determine the degree of differentiation. The tissue showed an upregulation of individual CD cell features as compared to embryonic epithelia in the neonatal kidney.

76 FR 17364 - Airworthiness Directives; Dassault-Aviation Model FALCON 7X Airplanes

Federal Register 2010, 2011, 2012, 2013, 2014

2011-03-29

... a negative distance to the ground despite the aircraft was flying at medium or high altitude. A... despite the aircraft was flying at medium or high altitude. A locked radio-altimeter 1 leads to untimely... certain abnormal conditions while the avionic system switches into landing mode during altitude cruise...

Reading on Paper and Digitally: What the Past Decades of Empirical Research Reveal

ERIC Educational Resources Information Center

Singer, Lauren M.; Alexander, Patricia A.

2017-01-01

This systematic literature review was undertaken primarily to examine the role that print and digitally mediums play in text comprehension. Overall, results suggest that medium plays an influential role under certain text or task conditions or for certain readers. Additional goals were to identify how researchers defined and measured…

EFFECT OF TEMPERATURE AND PORE SIZE ON THE HYDRAULIC PROPERTIES AND FLOW OF A HYDROCARBON OIL IN THE SUBSURFACE

EPA Science Inventory

Capillary pressure relationships in a porous medium determine the distribution of immiscible fluids under static conditions and can largely influence the movement of the fluids when the system is not at equilibrium. Theory predicts that for a given porous medium, the effect of di...

Using a Chart-table Medium to Focus Students' Attention on Science Concepts.

ERIC Educational Resources Information Center

Holliday, William G.; Benson, Garth D.

A study is presented which utilizes a selective attention model to investigate the learning effects of different questioning strategies under four experimental conditions using a non-prose medium (science textbook chart) containing information of varying established difficulty. A 14-row, 4-column chart describing 14 vitamins was used to present…

Production of novel antibiotics zeamines through optimizing Dickeya zeae fermentation conditions.

PubMed

Liao, Lisheng; Cheng, Yingying; Liu, Shiyin; Zhou, Jianuan; An, Shuwen; Lv, Mingfa; Chen, Yufan; Gu, Yanfang; Chen, Shaohua; Zhang, Lian-Hui

2014-01-01

Dickeya zeae strain EC1 was recently shown to produce a new type of phytotoxins designated as zeamine and zeamine II, which are potent wide-spectrum antibiotics against Gram-positive and Gram-negative bacterial pathogens, suggesting their promising potential as clinical medicines. In this study, the optimized medium composition and culture conditions for biosynthesis of novel antibiotics zeamines have been established by using response surface methodology, largely increasing the yield of zeamines from original about 7.35 µg · mL(-1) in minimal medium to about 150 µg · mL(-1) in LS5 medium. The study identified the major factors contributing to zeamines production, which include nitrate, sucrose, asparaginate, mineral elements Mg2+ and K+, and optimized amount of phosphate. In addition, the results showed that overexpression of zmsK in D. zeae strain EC1 could further increase zeamines yield to about 180 µg · mL(-1) in LS5 medium. The findings from this study could facilitate further characterization and utilization of these two novel antibiotics, and also provide useful clues for understanding the regulatory mechanisms that govern D. zeae virulence.

Cultured Human Retinal Pigment Epithelial (hRPE) Sheets: A Search for Suitable Storage Conditions.

PubMed

Khan, Ayyad Z; Utheim, Tor P; Reppe, Sjur; Sandvik, Leiv; Lyberg, Torstein; Roald, Borghild B-H; Ibrahim, Ibrahim B; Eidet, Jon R

2018-04-01

The advancement of human retinal pigment epithelial cell (hRPE) replacement therapy is partly dependent on optimization of cell culture, cell preservation, and storage medium. This study was undertaken to search for a suitable storage temperature and storage medium for hRPE. hRPE monolayer sheets were cultured under standard conditions at 37°C and then randomized for storage at six temperatures (4, 16, 20, 24, 28, and 37°C) for 7 days. After revealing a suitable storage temperature, hRPE sheets were subsequently stored with and without the silk protein sericin added to the storage medium. Live/dead assay, light microscopy, pH, and phenotypic expression of various proteins were used to assess cell cultures stored at different temperatures. After 7 days of storage, hRPE morphology was best preserved at 4°C. Addition of sericin to the storage medium maintained the characteristic morphology of the preserved cells, and improved pigmentation and levels of pigmentation-related proteins in the cultured hRPE sheets following a 7-day storage period at 4°C.

Optimization of Culture Medium Enhances Viable Biomass Production and Biocontrol Efficacy of the Antagonistic Yeast, Candida diversa.

PubMed

Liu, Jia; Li, Guangkun; Sui, Yuan

2017-01-01

Viable biomass production is a key determinant of suitability of antagonistic yeasts as potential biocontrol agents. This study investigated the effects of three metal ions (magnesium, ferrous, and zinc) on biomass production and viability of the antagonistic yeast, Candida diversa . Using response surface methodology to optimize medium components, a maximum biomass was obtained, when the collective Mg 2+ , Fe 2+ , and Zn 2+ concentrations were adjusted in a minimal mineral (MM) medium. Compared with the unmodified MM, and three ion-deficient MM media, yeast cells cultured in the three ion-modified MM medium exhibited a lower level of cellular oxidative damage, and a higher level of antioxidant enzyme activity. A biocontrol assay indicated that C. diversa grown in the ion-modified MM exhibited the greatest level of control of gray mold on apple fruit. These results provide new information on culture medium optimization to grow yeast antagonists in order to improve biomass production and biocontrol efficacy.

Optimization of Culture Medium Enhances Viable Biomass Production and Biocontrol Efficacy of the Antagonistic Yeast, Candida diversa

PubMed Central

Liu, Jia; Li, Guangkun; Sui, Yuan

2017-01-01

Viable biomass production is a key determinant of suitability of antagonistic yeasts as potential biocontrol agents. This study investigated the effects of three metal ions (magnesium, ferrous, and zinc) on biomass production and viability of the antagonistic yeast, Candida diversa. Using response surface methodology to optimize medium components, a maximum biomass was obtained, when the collective Mg2+, Fe2+, and Zn2+ concentrations were adjusted in a minimal mineral (MM) medium. Compared with the unmodified MM, and three ion-deficient MM media, yeast cells cultured in the three ion-modified MM medium exhibited a lower level of cellular oxidative damage, and a higher level of antioxidant enzyme activity. A biocontrol assay indicated that C. diversa grown in the ion-modified MM exhibited the greatest level of control of gray mold on apple fruit. These results provide new information on culture medium optimization to grow yeast antagonists in order to improve biomass production and biocontrol efficacy. PMID:29089939

Operating principles of Notch-Delta-Jagged module of cell-cell communication

NASA Astrophysics Data System (ADS)

Jolly, Mohit Kumar; Boareto, Marcelo; Lu, Mingyang; Onuchic, Jose' N.; Clementi, Cecilia; Ben-Jacob, Eshel

2015-05-01

Notch pathway is an evolutionarily conserved cell-cell communication mechanism governing cell-fate during development and tumor progression. It is activated when Notch receptor of one cell binds to either of its ligand—Delta or Jagged—of another cell. Notch-Delta (ND) signaling forms a two-way switch, and two cells interacting via ND signaling adopt different fates—Sender (high ligand, low receptor) and Receiver (low ligand, high receptor). Notch-Delta-Jagged signaling (NDJ) behaves as a three-way switch and enables an additional fate—hybrid Sender/Receiver (S/R) (medium ligand, medium receptor). Here, by extending our framework of NDJ signaling for a two-cell system, we show that higher production rate of Jagged, but not that of Delta, expands the range of parameters for which both cells attain the hybrid S/R state. Conversely, glycosyltransferase Fringe and cis-inhibition reduces this range of conditions, and reduces the relative stability of the hybrid S/R state, thereby promoting cell-fate divergence and consequently lateral inhibition-based patterns. Lastly, soluble Jagged drives the cells to attain the hybrid S/R state, and soluble Delta drives them to be Receivers. We also discuss the critical role of hybrid S/R state in promoting cancer metastasis by enabling collective cell migration and expanding cancer stem cell (CSC) population.

Modulation of medium-chain fatty acid synthesis in Synechococcus sp. PCC 7002 by replacing FabH with a Chaetoceros Ketoacyl-ACP synthase

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

Gu, Huiya; Jinkerson, Robert E.; Davies, Fiona K.

The isolation or engineering of algal cells synthesizing high levels of medium-chain fatty acids (MCFAs) is attractive to mitigate the high clouding point of longer chain fatty acids in algal based biodiesel. To develop a more informed understanding of MCFA synthesis in photosynthetic microorganisms, we isolated several algae from Great Salt Lake and screened this collection for MCFA accumulation to identify strains naturally accumulating high levels of MCFA. A diatom, Chaetoceros sp. GSL56, accumulated particularly high levels of C14 (up to 40%), with the majority of C14 fatty acids allocated in triacylglycerols. Using whole cell transcriptome sequencing and de novomore » assembly, putative genes encoding fatty acid synthesis enzymes were identified. Enzymes from this Chaetoceros sp. were expressed in the cyanobacterium Synechococcus sp. PCC 7002 to validate gene function and to determine whether eukaryotic enzymes putatively lacking bacterial evolutionary control mechanisms could be used to improve MCFA production in this promising production strain. Replacement of the Synechococcus 7002 native FabH with a Chaetoceros ketoacyl-ACP synthase Ill increased MCFA synthesis up to fivefold. In conclusion, the level of increase is dependent on promoter strength and culturing conditions.« less

Modulation of medium-chain fatty acid synthesis in Synechococcus sp. PCC 7002 by replacing FabH with a Chaetoceros Ketoacyl-ACP synthase

DOE PAGES

Gu, Huiya; Jinkerson, Robert E.; Davies, Fiona K.; ...

2016-05-26

The isolation or engineering of algal cells synthesizing high levels of medium-chain fatty acids (MCFAs) is attractive to mitigate the high clouding point of longer chain fatty acids in algal based biodiesel. To develop a more informed understanding of MCFA synthesis in photosynthetic microorganisms, we isolated several algae from Great Salt Lake and screened this collection for MCFA accumulation to identify strains naturally accumulating high levels of MCFA. A diatom, Chaetoceros sp. GSL56, accumulated particularly high levels of C14 (up to 40%), with the majority of C14 fatty acids allocated in triacylglycerols. Using whole cell transcriptome sequencing and de novomore » assembly, putative genes encoding fatty acid synthesis enzymes were identified. Enzymes from this Chaetoceros sp. were expressed in the cyanobacterium Synechococcus sp. PCC 7002 to validate gene function and to determine whether eukaryotic enzymes putatively lacking bacterial evolutionary control mechanisms could be used to improve MCFA production in this promising production strain. Replacement of the Synechococcus 7002 native FabH with a Chaetoceros ketoacyl-ACP synthase Ill increased MCFA synthesis up to fivefold. In conclusion, the level of increase is dependent on promoter strength and culturing conditions.« less

Effect of UV irradiation on organic matter extracted from treated Ohio River water studied through the use of electrospray mass spectrometry.

PubMed

Magnuson, Matthew L; Kelty, Catherine A; Sharpless, Charles M; Linden, Karl G; Fromme, William; Metz, Deborah H; Kashinkunti, Ramesh

2002-12-01

Ohio River water was treated by settling, sand filtration, and granular activated carbon filtration. It was then irradiated by low-pressure (monochromatic) and medium-pressure (polychromatic) UV lamps to investigate the effects of UV irradiation on the extracted organic matter (EOM). When the EOM, collected by solid phase extraction cartridges, was analyzed by conventional UV spectroscopy and size exclusion chromatography (SEC), no significant changes in the EOM were revealed for various UV doses. Positive and negative electrospray ionization mass spectrometry (ESI-MS) of the EOM produced mass spectra that vary significantly with UV dose. The UV dosage conditions also appear to affect the reactivity of the EOM to subsequent chlorination. The magnitude of the spectral changes is generally greater for medium-pressure lamps than for low pressure and increases with UV exposure. Based on the observed MS peaks, the changes may be due to the presence of lignin, resulting perhaps from photooxidation and/or photo rearrangement of macromolecules in the sample. When chlorination is used for secondary disinfection, these results suggest that it may be important to consider the effects of UV irradiation on the organic matter in the water before applying UV disinfection technology to a particular source water.

The Effect of Cryopreserved Human Placental Tissues on Biofilm Formation of Wound-Associated Pathogens.

PubMed

Mao, Yong; Singh-Varma, Anya; Hoffman, Tyler; Dhall, Sandeep; Danilkovitch, Alla; Kohn, Joachim

2018-01-08

Biofilm, a community of bacteria, is tolerant to antimicrobial agents and ubiquitous in chronic wounds. In a chronic DFU (Diabetic Foot Ulcers) clinical trial, the use of a human cryopreserved viable amniotic membrane (CVAM) resulted in a high rate of wound closure and reduction of wound-related infections. Our previous study demonstrated that CVAM possesses intrinsic antimicrobial activity against a spectrum of wound-associated bacteria under planktonic culture conditions. In this study, we evaluated the effect of CVAM and cryopreserved viable umbilical tissue (CVUT) on biofilm formation of S. aureus and P. aeruginosa , the two most prominent pathogens associated with chronic wounds. Firstly, we showed that, like CVAM, CVUT released antibacterial activity against multiple bacterial pathogens and the devitalization of CVUT reduced its antibacterial activity. The biofilm formation was then measured using a high throughput method and an ex vivo porcine dermal tissue model. We demonstrate that the formation of biofilm was significantly reduced in the presence of CVAM- or CVUT-derived conditioned media compared to control assay medium. The formation of P. aeruginosa biofilm on CVAM-conditioned medium saturated porcine dermal tissues was reduced 97% compared with the biofilm formation on the control medium saturated dermal tissues. The formation of S. auerus biofilm on CVUT-conditioned medium saturated dermal tissues was reduced 72% compared with the biofilm formation on the control tissues. This study is the first to show that human cryopreserved viable placental tissues release factors that inhibit biofilm formation. Our results provide an explanation for the in vivo observation of their ability to support wound healing.

Alternating air-medium exposure in rotating bioreactors optimizes cell metabolism in 3D novel tubular scaffold polyurethane foams.

PubMed

Tresoldi, Claudia; Stefani, Ilaria; Ferracci, Gaia; Bertoldi, Serena; Pellegata, Alessandro F; Farè, Silvia; Mantero, Sara

2017-04-26

In vitro dynamic culture conditions play a pivotal role in developing engineered tissue grafts, where the supply of oxygen and nutrients, and waste removal must be permitted within construct thickness. For tubular scaffolds, mass transfer is enhanced by introducing a convective flow through rotating bioreactors with positive effects on cell proliferation, scaffold colonization and extracellular matrix deposition. We characterized a novel polyurethane-based tubular scaffold and investigated the impact of 3 different culture configurations over cell behavior: dynamic (i) single-phase (medium) rotation and (ii) double-phase exposure (medium-air) rotation; static (iii) single-phase static culture as control. A new mixture of polyol was tested to create polyurethane foams (PUFs) as 3D scaffold for tissue engineering. The structure obtained was morphologically and mechanically analyzed tested. Murine fibroblasts were externally seeded on the novel porous PUF scaffold, and cultured under different dynamic conditions. Viability assay, DNA quantification, SEM and histological analyses were performed at different time points. The PUF scaffold presented interesting mechanical properties and morphology adequate to promote cell adhesion, highlighting its potential for tissue engineering purposes. Results showed that constructs under dynamic conditions contain enhanced viability and cell number, exponentially increased for double-phase rotation; under this last configuration, cells uniformly covered both the external surface and the lumen. The developed 3D structure combined with the alternated exposure to air and medium provided the optimal in vitro biochemical conditioning with adequate nutrient supply for cells. The results highlight a valuable combination of material and dynamic culture for tissue engineering applications.

Radiation quality dependence of signal transmission and bystander induced cell killing

NASA Astrophysics Data System (ADS)

Esposito, Giuseppe; Bertolotti, Alessia; Facoetti, Angelica; Grande, Sveva; Mariotti, Luca; Ottolenghi, Andrea; Ranza, Elena; Simone, Giustina; Sorrentino, Eugenio; Antonella Tabocchini, Maria

Low dose radiobiological studies have shown effects, observable in cells that are in the vicinity of irradiated cells, which are due to the release by irradiated cells of several cellular mediators among which Reactive Oxygen and Nitrogen Species (ROS, NRS), and cytokines are likely to play a key role. Despite the large number in the literature of studies on bystander effects induced by ionizing radiation the results are still conflicting, and further studies are therefore needed on the possible underlying mechanisms. The dependence on radiation quality deserve particular attention because bystander mechanisms are probably more important with high-LET irradi-ations, where many cells are not hit (bystander). Moreover, due to the different patterns of energy deposition, the cellular response to low LET and high LET radiation can be different. Understanding whether these cells can contribute to the adverse effects of low radiation doses in a radiation quality-dependent fashion might have important implications in risk estimates for both cancer induction and non-cancer diseases. In this context, we addressed to the study of the bystander induced cell killing after incubation with "conditioned medium" from primary human fibroblasts irradiated with 0.1 and 0.5 Gy of α-particles or γ-rays. Medium transfer was performed after 1h incubation from irradiation. The results have confirmed a reduction in clonogenic survival after incubation with medium from α-irradiated cells, independently of the dose; similar results were obtained after γ-irradiation, although in this case a slight dose depen-dence could be envisaged. Interleukin-6 (IL-6) and Interleukin-8 (IL-8) levels were measured in the conditioned medium collected up to 20 hours after irradiation with α-particles and γ-rays in the dose-range of 0.1-1.0 Gy, in parallel with evaluation of their receptor expression in irradi-ated and bystander cells. Concerning IL-6, we observed the strongest modulation of its release at 20 hours post exposure, whereas IL-8 release was significantly increased at shorter times, i.e. 5-7 hours after irradiation. The expression of their receptors was modulated in both irradiated and bystander cells, although it is not apparently correlated with the relative interleukin re-lease. In order to investigate possible correlation between NRS and cytokines as early and late mediators in the signalling chain leading to bystander induced cell killing, experiments were performed using c-PTIO, a well known scavengers of RNS. In these experiments conditioned medium taken after 1h or 5h from α-particle irradiated cells was used. The results obtained after 5h in the absence of c-PTIO didn't show any further decrease in clonogenic survival of bystander cells. The presence of the scavenger seems to reduce, the bystander induced cell killing indicating that RNS are involved in the transduction of the bystander signal. Experi-ments, performed with both c-PTIO and DMSO, a scavenger of hydroxyl radicals, showed that RNS and ROS play a role in some cytokine pathways (interleukins release and their receptor expression) activated by irradiation. ACKNOWLEDGEMENTS This work was partially supported by the European Commission (EC Contract FP6-36465, "NOTE")

Optimization of cultivation conditions of fermented shaggy ink cap culinary-medicinal mushroom, Coprinus comatus (O.Mull.:Fr.) Pers. (higher Basidiomycetes) rich in Vanadium.

PubMed

Zhang, Chunjing; Qi, Xiaodan; Shi, Yan; Sun, Yan; Li, Shuyan; Gao, Xiulan; Yu, Haitao

2012-01-01

The present paper is mainly aimed at optimization of cultivation conditions of fermented mushrooms of Coprinus comatus rich in vanadium (CCRV). Initial screening of effects of carbon source, temperature, pH, and inoculum size were done by using a one-factor-at-a-time method. The results obtained in that study showed that the optimal medium composition was 30 g glucose/Lin YEPG medium, initial pH 6.0, inoculum volume 10%, and incubation time 120 h. Then the medium was subjected to screening of the most significant parameters using the L9 orthogonal array to solve multivariable equations simultaneously. The results obtained in this study showed that the optimal medium composition was 0.4% V and 30 g glucose/Lin YEPG medium, initial pH 5.0, inoculum volume 15%, and incubation time 120 h. At this medium composition, the mycelial biomass and V content were 7.18 ± 0.24 g/L and 3786.0 ± 17 μg/g, respectively. The anti-diabetic potential of CCRV produced with the optimal level was tested in alloxan-induced diabetes. After the mice were administered (i.g.) with CCRV, the level of blood sugar in the CCRV group was very close to that of the control group. These findings suggested that CCRV produced with the optimal level is useful in the control of diabetes mellitus.

Near-IR laser-triggered target cell collection using a carbon nanotube-based cell-cultured substrate.

PubMed

Sada, Takao; Fujigaya, Tsuyohiko; Niidome, Yasuro; Nakazawa, Kohji; Nakashima, Naotoshi

2011-06-28

Unique near-IR optical properties of single-walled carbon nanotube (SWNTs) are of interest in many biological applications. Here we describe the selective cell detachment and collection from an SWNT-coated cell-culture dish triggered by near-IR pulse laser irradiation. First, HeLa cells were cultured on an SWNT-coated dish prepared by a spraying of an aqueous SWNT dispersion on a glass dish. The SWNT-coated dish was found to show a good cell adhesion behavior as well as a cellular proliferation rate similar to a conventional glass dish. We discovered, by near-IR pulse laser irradiation (at the laser power over 25 mW) to the cell under optical microscopic observation, a quick single-cell detachment from the SWNT-coated surface. Shockwave generation from the irradiated SWNTs is expected to play an important role for the cell detachment. Moreover, we have succeeded in catapulting the target single cell from the cultured medium when the depth of the medium was below 150 μm and the laser power was stronger than 40 mW. The captured cell maintained its original shape. The retention of the genetic information of the cell was confirmed by the polymerase chain reaction (PCR) technique. A target single-cell collection from a culture medium under optical microscopic observation is significant in wide fields of single-cell studies in biological areas.

Unexpected Link between Metal Ion Deficiency and Autophagy in Aspergillus fumigatus▿ †

PubMed Central

Richie, Daryl L.; Fuller, Kevin K.; Fortwendel, Jarrod; Miley, Michael D.; McCarthy, Jason W.; Feldmesser, Marta; Rhodes, Judith C.; Askew, David S.

2007-01-01

Autophagy is the major cellular pathway for bulk degradation of cytosolic material and is required to maintain viability under starvation conditions. To determine the contribution of autophagy to starvation stress responses in the filamentous fungus Aspergillus fumigatus, we disrupted the A. fumigatus atg1 gene, encoding a serine/threonine kinase required for autophagy. The ΔAfatg1 mutant showed abnormal conidiophore development and reduced conidiation, but the defect could be bypassed by increasing the nitrogen content of the medium. When transferred to starvation medium, wild-type hyphae were able to undergo a limited amount of growth, resulting in radial expansion of the colony. In contrast, the ΔAfatg1 mutant was unable to grow under these conditions. However, supplementation of the medium with metal ions rescued the ability of the ΔAfatg1 mutant to grow in the absence of a carbon or nitrogen source. Depleting the medium of cations by using EDTA was sufficient to induce autophagy in wild-type A. fumigatus, even in the presence of abundant carbon and nitrogen, and the ΔAfatg1 mutant was severely growth impaired under these conditions. These findings establish a role for autophagy in the recycling of internal nitrogen sources to support conidiophore development and suggest that autophagy also contributes to the recycling of essential metal ions to sustain hyphal growth when exogenous nutrients are scarce. PMID:17921348

Characteristic time scales for diffusion processes through layers and across interfaces

NASA Astrophysics Data System (ADS)

Carr, Elliot J.

2018-04-01

This paper presents a simple tool for characterizing the time scale for continuum diffusion processes through layered heterogeneous media. This mathematical problem is motivated by several practical applications such as heat transport in composite materials, flow in layered aquifers, and drug diffusion through the layers of the skin. In such processes, the physical properties of the medium vary across layers and internal boundary conditions apply at the interfaces between adjacent layers. To characterize the time scale, we use the concept of mean action time, which provides the mean time scale at each position in the medium by utilizing the fact that the transition of the transient solution of the underlying partial differential equation model, from initial state to steady state, can be represented as a cumulative distribution function of time. Using this concept, we define the characteristic time scale for a multilayer diffusion process as the maximum value of the mean action time across the layered medium. For given initial conditions and internal and external boundary conditions, this approach leads to simple algebraic expressions for characterizing the time scale that depend on the physical and geometrical properties of the medium, such as the diffusivities and lengths of the layers. Numerical examples demonstrate that these expressions provide useful insight into explaining how the parameters in the model affect the time it takes for a multilayer diffusion process to reach steady state.

Characteristic time scales for diffusion processes through layers and across interfaces.

PubMed

Carr, Elliot J

2018-04-01

This paper presents a simple tool for characterizing the time scale for continuum diffusion processes through layered heterogeneous media. This mathematical problem is motivated by several practical applications such as heat transport in composite materials, flow in layered aquifers, and drug diffusion through the layers of the skin. In such processes, the physical properties of the medium vary across layers and internal boundary conditions apply at the interfaces between adjacent layers. To characterize the time scale, we use the concept of mean action time, which provides the mean time scale at each position in the medium by utilizing the fact that the transition of the transient solution of the underlying partial differential equation model, from initial state to steady state, can be represented as a cumulative distribution function of time. Using this concept, we define the characteristic time scale for a multilayer diffusion process as the maximum value of the mean action time across the layered medium. For given initial conditions and internal and external boundary conditions, this approach leads to simple algebraic expressions for characterizing the time scale that depend on the physical and geometrical properties of the medium, such as the diffusivities and lengths of the layers. Numerical examples demonstrate that these expressions provide useful insight into explaining how the parameters in the model affect the time it takes for a multilayer diffusion process to reach steady state.

Low-cost production of 6G-fructofuranosidase with high value-added astaxanthin by Xanthophyllomyces dendrorhous.

PubMed

Ning, Yawei; Li, Qiang; Chen, Feng; Yang, Na; Jin, Zhengyu; Xu, Xueming

2012-01-01

The effects of medium composition and culture conditions on the production of (6)G-fructofuranosidase with value-added astaxanthin were investigated to reduce the capital cost of neo-fructooligosaccharides (neo-FOS) production by Xanthophyllomyces dendrorhous. The sucrose and corn steep liquor (CSL) were found to be the optimal carbon source and nitrogen source, respectively. CSL and initial pH were selected as the critical factors using Plackett-Burman design. Maximum (6)G-fructofuranosidase 242.57 U/mL with 5.23 mg/L value-added astaxanthin was obtained at CSL 52.5 mL/L and pH 7.89 by central composite design. Neo-FOS yield could reach 238.12 g/L under the optimized medium conditions. Cost analysis suggested 66.3% of substrate cost was reduced compared with that before optimization. These results demonstrated that the optimized medium and culture conditions could significantly enhance the production of (6)G-fructofuranosidase with value-added astaxanthin and remarkably decrease the substrate cost, which opened up possibilities to produce neo-FOS industrially. Copyright © 2011 Elsevier Ltd. All rights reserved.

Use of spent mushroom substrate for production of Bacillus thuringiensis by solid-state fermentation.

PubMed

Wu, Songqing; Lan, Yanjiao; Huang, Dongmei; Peng, Yan; Huang, Zhipeng; Xu, Lei; Gelbic, Ivan; Carballar-Lejarazu, Rebeca; Guan, Xiong; Zhang, Lingling; Zou, Shuangquan

2014-02-01

The aim of this study was to explore a cost-effective method for the mass production of Bacillus thuringiensis (Bt) by solid-state fermentation. As a locally available agroindustrial byproduct, spent mushroom substrate (SMS) was used as raw material for Bt cultivation, and four combinations of SMS-based media were designed. Fermentation conditions were optimized on the best medium and the optimal conditions were determined as follows: temperature 32 degrees C, initial pH value 6, moisture content 50%, the ratio of sieved material to initial material 1:3, and inoculum volume 0.5 ml. Large scale production of B. thuringiensis subsp. israelensis (Bti) LLP29 was conducted on the optimal medium at optimal conditions. High toxicity (1,487 international toxic units/milligram) and long larvicidal persistence of the product were observed in the study, which illustrated that SMS-based solid-state fermentation medium was efficient and economical for large scale industrial production of Bt-based biopesticides. The cost of production of 1 kg of Bt was approximately US$0.075.

Lignolytic enzymes produced by Trametes villosa ccb176 under different culture conditions

PubMed Central

Yamanaka, Renata; Soares, Clarissa F.; Matheus, Dácio R.; Machado, Kátia M.G.

2008-01-01

The expression of the enzymatic system produced by basidiomycetous fungi, which is involved in the degradation of xenobiotics, mainly depends on culture conditions, especially of the culture medium composition. Trametes villosa is a strain with a proven biotechnological potential for the degradation of organochlorine compounds and for the decolorization of textile dyes. The influence of glucose concentration, addition of a vegetable oil-surfactant emulsion, nature of the surfactant and the presence of manganese and copper on the growth, pH and production of laccase, total peroxidase and manganese-dependent peroxidase activities were evaluated. In general, acidification of the medium was observed, with the pH reaching a value close to 3.5 within the first days of growth. Laccase was the main activity detected under the different conditions and was produced throughout the culture period of the fungus, irrespective of the growth phase. Supplementation of the medium with vegetable oil emulsified with a surfactant induced manganese-dependent peroxidase activity in T. villosa. Higher specific yields of laccase activity were obtained with the addition of copper. PMID:24031184

Modeling the growth of Lactobacillus viridescens under non-isothermal conditions in vacuum-packed sliced ham.

PubMed

Silva, Nathália Buss da; Longhi, Daniel Angelo; Martins, Wiaslan Figueiredo; Laurindo, João Borges; Aragão, Gláucia Maria Falcão de; Carciofi, Bruno Augusto Mattar

2017-01-02

Lactic acid bacteria (LAB) are responsible for spoiling vacuum-packed meat products, such as ham. Since the temperature is the main factor affecting the microbial dynamic, the use of mathematical models describing the microbial behavior into a non-isothermal environment can be very useful for predicting food shelf life. In this study, the growth of Lactobacillus viridescens was measured in vacuum-packed sliced ham under non-isothermal conditions, and the predictive ability of primary (Baranyi and Roberts, 1994) and secondary (Square Root) models were assessed using parameters estimated in MRS culture medium under isothermal conditions (between 4 and 30°C). Fresh ham piece was sterilized, sliced, inoculated, vacuum-packed, and stored in a temperature-controlled incubator at five different non-isothermal conditions (between 4 and 25°C) and one isothermal condition (8°C). The mathematical models obtained in MRS medium were assessed by comparing predicted values with L. viridescens growth data in vacuum-packed ham. Its predictive ability was assessed through statistical indexes, with good results (bias factor between 0.95 and 1.03; accuracy factor between 1.04 and 1.07, and RMSE between 0.76 and 1.33), especially in increasing temperature, which predictions were safe. The model parameters obtained from isothermal growth data in MRS medium enabled to estimate the shelf life of a commercial ham under non-isothermal conditions in the temperature range analyzed. Copyright © 2016 Elsevier B.V. All rights reserved.

IGF-I and NEFA concentrations in fetal fluids of term pregnancy dogs.

PubMed

Meloni, Tea; Comin, Antonella; Rota, Alessandro; Peric, Tanja; Contri, Alberto; Veronesi, Maria Cristina

2014-06-01

Insulin-like growth factor-I (IGF-I) and non-esterified fatty acids (NEFA) play an essential role in fetal growth and development. To date, fetal fluids IGF-I and NEFA levels at term canine pregnancy are unknown and could be related to the neonatal development and breed size. For these reasons, the aims of the present study were as follows: (1) to evaluate IGF-I and NEFA concentrations in fetal fluids collected from normally developed and viable newborn puppies born at term of normal pregnancies; (2) to assess possible differences between IGF-I and NEFA levels in amniotic compared with allantoic fluid; (3) to detect possible relationship between breed body size and IGF-I and NEFA amniotic and allantoic concentrations; (4) to evaluate possible differences in IGF-I fetal fluids levels between male and female puppies; and (5) to assess possible correlations between the two hormones in each type of fluid. The study enrolled 25 pure breed bitches submitted to elective Cesarean section at term because of the high risk of dystocia or previous troubles at parturition. At surgery, amniotic and allantoic fluids were collected and assayed for IGF-I and NEFA. IGF-I and NEFA amounts in both amniotic and allantoic fluids of different breed size bitches (small: ≤10 kg; medium: 11-25 kg; large: 26-40 kg) were detected, as well as the effect of gender on IGF-I levels. On a total of 73 amniotic and 76 allantoic samples collected by normal, viable, and mature newborns, the mean IGF-I concentration was significantly higher in amniotic than in allantoic fluid in all three groups, but the amniotic IGF-I levels were significantly lower in small and medium size bitches when compared with large ones. No significant differences were found in allantoic IGF-I concentrations among size groups. A significant effect of the puppy gender on IGF-I content in both fetal fluids was not reported. Regarding NEFA, in all the three groups, the mean NEFA concentration did not significantly differ between amnion and allantois, but in both fetal fluids, higher NEFA levels were detected in samples belonging to small breeds when compared with medium and large. These data strongly indicated that, also in the dog, a relation between fetal fluids IGF-I and NEFA concentrations and breed size exists. Further research is needed to elucidate the possible role of IGF-I and NEFA in the pathologic conditions related to canine fetal growth. Copyright © 2014 Elsevier Inc. All rights reserved.

Probing the mechanisms of drug release from amorphous solid dispersions in medium-soluble and medium-insoluble carriers.

PubMed

Sun, Dajun D; Lee, Ping I

2015-08-10

The objective of the current study is to mechanistically differentiate the dissolution and supersaturation behaviors of amorphous drugs from amorphous solid dispersions (ASDs) based on medium-soluble versus medium-insoluble carriers under nonsink dissolution conditions through a direct head-to-head comparison. ASDs of indomethacin (IND) were prepared in several polymers which exhibit different solubility behaviors in acidic (pH1.2) and basic (pH7.4) dissolution media. The selected polymers range from water-soluble (e.g., PVP and Soluplus) and water-insoluble (e.g., ethylcellulose and Eudragit RL PO) to those only soluble in an acidic or basic dissolution medium (e.g., Eudragit E100, Eudragit L100, and HPMCAS). At 20wt.% drug loading, DSC and powder XRD analysis confirmed that the majority of incorporated IND was present in an amorphous state. Our nonsink dissolution results confirm that whether the carrier matrix is medium soluble determines the release mechanism of amorphous drugs from ASD systems which has a direct impact on the rate of supersaturation generation, thus in turn affecting the evolution of supersaturation in amorphous systems. For example, under nonsink dissolution conditions, the release of amorphous IND from medium-soluble carriers is governed by a dissolution-controlled mechanism leading to an initial surge of supersaturation followed by a sharp decline in drug concentration due to rapid nucleation and crystallization. In contrast, the dissolution of IND ASD from medium-insoluble carriers is more gradual as drug release is regulated by a diffusion-controlled mechanism by which drug supersaturation is built up gradually and sustained over an extended period of time without any apparent decline. Since several tested carrier polymers can be switched from soluble to insoluble by simply changing the pH of the dissolution medium, the results obtained here provide unequivocal evidence of the proposed transition of kinetic solubility profiles from the same ASD system induced by changes in the drug release mechanism in dissolution medium of a different pH. Copyright © 2015 Elsevier B.V. All rights reserved.

26 CFR 31.3102-1 - Collection of, and liability for, employee tax; in general.

Code of Federal Regulations, 2010 CFR

2010-04-01

... exclusion from wages of remuneration paid in any medium other than cash for certain types of services, see § 31.3121(a)(7)-1, relating to such remuneration paid for service not in the course of the employer's..., relating to such remuneration paid for agricultural labor.) For provisions relating to the collection of...

Internet-Mediated Technologies and Mixed Methods Research: Problems and Prospects

ERIC Educational Resources Information Center

Hesse-Biber, Sharlene; Griffin, Amy J.

2013-01-01

This article provides an examination of a range of mixed methods research projects that employ Internet-mediated technologies (IMT) for data collection. Using a case study approach, this article allows for the uncovering of a process by which IMT are used as a data collection medium in mixed methods praxis. Under the theoretical position of medium…

MD PHEV/EV ARRA Project Data Collection and Reporting (Presentation)

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

Walkowicz, K.; Ramroth, L.; Duran, A.

2012-01-01

This presentation describes a National Renewable Energy Laboratory project to collect and analyze commercial fleet deployment data from medium-duty plug-in hybrid electric and all-electric vehicles that were deployed using funds from the American Recovery and Reinvestment Act. This work supports the Department of Energy's Vehicle Technologies Program and its Advanced Vehicle Testing Activity.

Cross Modulation of Two Laser Beams at the Individual-Photon Level

DTIC Science & Technology

2014-09-12

medium, such that the photons travel as slow-light polaritons [15,25,26], whose atomic excitation component can block the transmission of another light...through the ensemble, traveling in the medium as slow-light polaritons , a superposition of a photon and a collective atomic excitation to the state...unclassified Standard Form 298 (Rev. 8-98) Prescribed by ANSI Std Z39-18 slow-light polariton , the polariton’s atomic component in state jci reduces

Influence of the Culture Medium in Dose-Response Effect of the Chlorhexidine on Streptococcus mutans Biofilms

PubMed Central

de Queiroz, Vanessa Salvadego; Ccahuana-Vásquez, Renzo Alberto; Tedesco, Alcides Fabiano; Lyra, Luzia; Cury, Jaime Aparecido; Schreiber, Angélica Zaninelli

2016-01-01

The aim of this study was to evaluate the influence of culture medium on dose-response effect of chlorhexidine (CHX) on Streptococcus mutans UA159 biofilm and validate the use of the cation-adjusted-Müller-Hinton broth (MH) for the evaluation of antibacterial activity. Ultrafiltered Tryptone-Yeast Extract Broth (UTYEB) was compared against MH and MH with blood supplementation (MHS). For each medium, six groups (n = 4) were assessed: two negative control groups (baseline 48 and 120 h) and four experimental groups (0.0001, 0.001, 0.012, and 0.12% CHX). S. mutans biofilm grew on glass slides of each media containing 1% sucrose. After 48 h of growth, biofilms of baseline 48 h were collected and the other groups were treated for 1 min, twice a day, for 3 days, with their respective treatments. The media were changed daily and pH was measured. After 120 h, biofilms were collected and dry weight and viable microorganisms were determined. Results showed CHX dose-response effect being observed in all media for all the variables. However, MH and MHS showed higher sensitivity than UTYEB (p < 0.05). We can conclude that the culture medium does influence dose-response effect of CHX on Streptococcus mutans biofilm and that MH can be used for antibacterial activity. PMID:27293967

Filterability of the suspension from germanium precipitation with aqueous tannin extract solution

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

Mikhailov, N.F.; Petropol'skii, V.M.; Semenenko, L.E.

1978-01-01

We have already described the use of a neutral aqueous solution of tannin extract to recover germanium from collecting-mains liquor in coking plants. Further pilot commercial trials have encountered problems with the poor filterability of the precipitate obtained when germanium is extracted with this reagent in alkaline media. There are published references to the colloidal nature of the precipitated tannin-germanium complex. It is also known that the alkalinity of the medium influences the degree of association in colloidal systems to a marked extent. Accordingly, special research was needed to establish the relationship between the pH of the precipitation medium andmore » the filterability of the germanium deposit. Samples of collecting-mains liquor were taken from one of the southern coking plants to determine the optimum filtration behavior. The collecting-mains liquor should first be purged of volatile ammonia and then adjusted to pH = 6.5 to 6.7 for precipitation.« less

Dose accuracy of a durable insulin pen with memory function, before and after simulated lifetime use and under stress conditions.

PubMed

Xue, Ligang; Mikkelsen, Kristian Handberg

2013-03-01

The objective of this study was to assess the dose accuracy of NovoPen® 5 in delivering low, medium and high doses of insulin before and after simulated lifetime use. A secondary objective was to evaluate the durability of the pen and its memory function under various stress conditions designed to simulate conditions that may be encountered in everyday use of an insulin pen. All testing was conducted according to International Organization for Standardization guideline 11608-1, 2000 for pen injectors. Dose accuracy was measured for the delivery of 1 unit (U) (10 mg), 30 U (300 mg) and 60 U (600 mg) test medium in standard, cool and hot conditions and before and after simulated lifetime use. Dose accuracy was also tested after preconditioning in dry heat storage; cold storage; damp cyclical heat; shock, bump and vibration; free fall and after electrostatic charge and radiated field test. Memory function was tested under all temperature and physical conditions. NovoPen 5 maintained dosing accuracy and memory function at minimum, medium and maximum doses in standard, cool and hot conditions, stress tests and simulated lifetime use. The pens remained intact and retained dosing accuracy and a working memory function at all doses after exposure to variations in temperature and after physical challenge. NovoPen 5 was accurate at all doses tested and under various functionality tests. Its durable design ensured that the dose accuracy and memory function were retained under conditions of stress likely to be encountered in everyday use.

Thermal non-equilibrium in porous medium adjacent to vertical plate: ANN approach

NASA Astrophysics Data System (ADS)

Ahmed, N. J. Salman; Ahamed, K. S. Nazim; Al-Rashed, Abdullah A. A. A.; Kamangar, Sarfaraz; Athani, Abdulgaphur

2018-05-01

Thermal non-equilibrium in porous medium is a condition that refers to temperature discrepancy in solid matrix and fluid of porous medium. This type of flow is complex flow requiring complex set of partial differential equations that govern the flow behavior. The current work is undertaken to predict the thermal non-equilibrium behavior of porous medium adjacent to vertical plate using artificial neural network. A set of neurons in 3 layers are trained to predict the heat transfer characteristics. It is found that the thermal non-equilibrium heat transfer behavior in terms of Nusselt number of fluid as well as solid phase can be predicted accurately by using well-trained neural network.

Transverse Vibration of Tapered Single-Walled Carbon Nanotubes Embedded in Viscoelastic Medium

NASA Astrophysics Data System (ADS)

Lei, Y. J.; Zhang, D. P.; Shen, Z. B.

2017-12-01

Based on the nonlocal theory, Euler-Bernoulli beam theory and Kelvin viscoelastic foundation model, free transverse vibration is studied for a tapered viscoelastic single-walled carbon nanotube (visco-SWCNT) embedded in a viscoelastic medium. Firstly, the governing equations for vibration analysis are established. And then, we derive the natural frequencies in closed form for SWCNTs with arbitrary boundary conditions by applying transfer function method and perturbation method. Numerical results are also presented to discuss the effects of nonlocal parameter, relaxation time and taper parameter of SWCNTs, and material property parameters of the medium. This study demonstrates that the proposed model is available for vibration analysis of the tapered SWCNTs-viscoelastic medium coupling system.

Characterization of release of basic fibroblast growth factor from bovine retinal endothelial cells in monolayer cultures.

PubMed Central

Brooks, R A; Burrin, J M; Kohner, E M

1991-01-01

Release of basic fibroblast growth factor (bFGF) was investigated in bovine retinal endothelial cells (BREC) maintained in monolayer culture. Confluent cells released bFGF into serum-free culture medium or medium containing 5% serum at rates of up to 105.2 and 61.3 pM/day respectively. bFGF release coincided with a decrease in monolayer cell number and increases in lactate dehydrogenase (LDH) concentration and cells and cell-debris particles in the medium, which suggested that cell damage and lysis were responsible for growth-factor release. Maximum bFGF release at 24 h (230 +/- 10 pM) occurred when the cells were treated with lipopolysaccharide (10 micrograms/ml), which also produced the greatest changes in parameters of cell damage. Sub-confluent cells showed little overt damage at 24 h, but released bFGF (78 +/- 20 pM) along with LDH, indicating that some cell lysis had occurred. Insulin-like growth factor 1 (IGF-1) was also released into serum-free culture medium at a rate of 0.34 nM/day, but not into medium containing serum or when the cells were treated with lipopolysaccharide. This implies that the mechanism of IGF-1 release is different from that of bFGF and is not related to cell damage. Culture medium conditioned by BREC stimulated the proliferation of these cells, as measured by an increase in their incorporation of [methyl-3H]thymidine from 7550 +/- 479 to 10467 +/- 924 d.p.m. These results demonstrate that bFGF is released from damaged BREC and that medium conditioned by these cells can stimulate retinal-endothelial-cell proliferation. This strengthens the case for an involvement of this growth factor in retinal neovascularization. Images Fig. 1. PMID:2039465

Medium-Chain Fatty Acids Improve Cognitive Function in Intensively Treated Type 1 Diabetic Patients and Support In Vitro Synaptic Transmission During Acute Hypoglycemia

PubMed Central

Page, Kathleen A.; Williamson, Anne; Yu, Namyi; McNay, Ewan C.; Dzuira, James; McCrimmon, Rory J.; Sherwin, Robert S.

2009-01-01

OBJECTIVE We examined whether ingestion of medium-chain triglycerides could improve cognition during hypoglycemia in subjects with intensively treated type 1 diabetes and assessed potential underlying mechanisms by testing the effect of β-hydroxybutyrate and octanoate on rat hippocampal synaptic transmission during exposure to low glucose. RESEARCH DESIGN AND METHODS A total of 11 intensively treated type 1 diabetic subjects participated in stepped hyperinsulinemic- (2 mU · kg−1 · min−1) euglycemic- (glucose ∼5.5 mmol/l) hypoglycemic (glucose ∼2.8 mmol/l) clamp studies. During two separate sessions, they randomly received either medium-chain triglycerides or placebo drinks and performed a battery of cognitive tests. In vitro rat hippocampal slice preparations were used to assess the ability of β-hydroxybutyrate and octanoate to support neuronal activity when glucose levels are reduced. RESULTS Hypoglycemia impaired cognitive performance in tests of verbal memory, digit symbol coding, digit span backwards, and map searching. Ingestion of medium-chain triglycerides reversed these effects. Medium-chain triglycerides also produced higher free fatty acids and β-hydroxybutyrate levels compared with placebo. However, the increase in catecholamines and symptoms during hypoglycemia was not altered. In hippocampal slices β-hydroxybutyrate supported synaptic transmission under low-glucose conditions, whereas octanoate could not. Nevertheless, octanoate improved the rate of recovery of synaptic function upon restoration of control glucose concentrations. CONCLUSIONS Medium-chain triglyceride ingestion improves cognition without adversely affecting adrenergic or symptomatic responses to hypoglycemia in intensively treated type 1 diabetic subjects. Medium-chain triglycerides offer the therapeutic advantage of preserving brain function under hypoglycemic conditions without causing deleterious hyperglycemia. PMID:19223595

Applications systems verification and transfer project. Volume 2: Operational applications of satellite snow-cover observations and data-collection systems in the Arizona test site

NASA Technical Reports Server (NTRS)

Schumann, H. H.

1981-01-01

Ground surveys and aerial observations were used to monitor rapidly changing moisture conditions in the Salt-Verde watershed. Repetitive satellite snow cover observations greatly reduce the necessity for routine aerial snow reconnaissance flights over the mountains. High resolution, multispectral imagery provided by LANDSAT satellite series enabled rapid and accurate mapping of snow-cover distributions for small- to medium-sized subwatersheds; however, the imagery provided only one observation every 9 days of about a third of the watershed. Low resolution imagery acquired by the ITOSa dn SMS/GOES meteorological satellite series provides the daily synoptic observation necessary to monitor the rapid changes in snow-covered area in the entire watershed. Short term runoff volumes can be predicted from daily sequential snow cover observations.

Ejecta- and Size-Scaling Considerations from Impacts of Glass Projectiles into Sand

NASA Technical Reports Server (NTRS)

Anderson J. L. B.; Cintala, M. J.; Siebenaler, S. A.; Barnouin-Jha, O. S.

2007-01-01

One of the most promising means of learning how initial impact conditions are related to the processes leading to the formation of a planetary-scale crater is through scaling relationships.1,2,3 The first phase of deriving such relationships has led to great insight into the cratering process and has yielded predictive capabilities that are mathematically rigorous and internally consistent. Such derivations typically have treated targets as continuous media; in many, cases, however, planetary materials represent irregular and discontinuous targets, the effects of which on the scaling relationships are still poorly understood.4,5 We continue to examine the effects of varying impact conditions on the excavation and final dimensions of craters formed in sand. Along with the more commonly treated variables such as impact speed, projectile size and material, and impact angle,6 such experiments also permit the study of changing granularity and friction angle of the target materials. This contribution presents some of the data collected during and after the impact of glass spheres into a medium-grained sand.

Albumin Enhances Caspofungin Activity against Aspergillus Species by Facilitating Drug Delivery to Germinating Hyphae

PubMed Central

Ioannou, Petros; Andrianaki, Aggeliki; Akoumianaki, Tonia; Kyrmizi, Irene; Albert, Nathaniel; Perlin, David; Samonis, George

2015-01-01

The modest in vitro activity of echinocandins against Aspergillus implies that host-related factors augment the action of these antifungal agents in vivo. We found that, in contrast to the other antifungal agents (voriconazole, amphotericin B) tested, caspofungin exhibited a profound increase in activity against various Aspergillus species under conditions of cell culture growth, as evidenced by a ≥4-fold decrease in minimum effective concentrations (MECs) (P = 0. 0005). Importantly, the enhanced activity of caspofungin against Aspergillus spp. under cell culture conditions was strictly dependent on serum albumin and was not observed with the other two echinocandins, micafungin and anidulafungin. Of interest, fluorescently labeled albumin bound preferentially on the surface of germinating Aspergillus hyphae, and this interaction was further enhanced upon treatment with caspofungin. In addition, supplementation of cell culture medium with albumin resulted in a significant, 5-fold increase in association of fluorescently labeled caspofungin with Aspergillus hyphae (P < 0.0001). Collectively, we found a novel synergistic interaction between albumin and caspofungin, with albumin acting as a potential carrier molecule to facilitate antifungal drug delivery to Aspergillus hyphae. PMID:26643329

Simplified pulse reactor for real-time long-term in vitro testing of biological heart valves.

PubMed

Schleicher, Martina; Sammler, Günther; Schmauder, Michael; Fritze, Olaf; Huber, Agnes J; Schenke-Layland, Katja; Ditze, Günter; Stock, Ulrich A

2010-05-01

Long-term function of biological heart valve prostheses (BHV) is limited by structural deterioration leading to failure with associated arterial hypertension. The objective of this work was development of an easy to handle real-time pulse reactor for evaluation of biological and tissue engineered heart valves under different pressures and long-term conditions. The pulse reactor was made of medical grade materials for placement in a 37 degrees C incubator. Heart valves were mounted in a housing disc moving horizontally in culture medium within a cylindrical culture reservoir. The microprocessor-controlled system was driven by pressure resulting in a cardiac-like cycle enabling competent opening and closing of the leaflets with adjustable pulse rates and pressures between 0.25 to 2 Hz and up to 180/80 mmHg, respectively. A custom-made imaging system with an integrated high-speed camera and image processing software allow calculation of effective orifice areas during cardiac cycle. This simple pulse reactor design allows reproducible generation of patient-like pressure conditions and data collection during long-term experiments.

Analyses of Failure Mechanisms in Woven Graphite/Polyimide Composites with Medium and High Modulus Graphite Fibers Subjected to In-Plane Shear

NASA Technical Reports Server (NTRS)

Kumosa, M.; Armentrout, D.; Rupnowski, P.; Kumosa, L.; Shin, E.; Sutter, J. K.

2003-01-01

The application of the Iosipescu shear test for the room and high temperature failure analyses of the woven graphite/polyimide composites with the medium (T-650) and igh (M40J and M60J) modulus graphite fibers is discussed. The M40J/PMR-II-50 and M60J/PMR-II-50 composites were tested as supplied and after thermal conditioning. The effect of temperature and conditioning on the initiation of intralaminar damage and the shear strength of the composites was established.

Stimulation of ATP synthesis in Halobacterium halobium R1 by light-induced or artifically created proton electrochemical potential gradients across the cell membrane.

PubMed

Danon, A; Caplan, S R

1976-01-15

The relationship between proton movement and phosphorylation in Halo-bacterium halobium R1 has been investigated under anaerobic conditions. The light-induced changes in the bacteriorhodopsin are accompanied by proton movements across the membrane which result in pH changes in the suspending medium. The initial alkaline shift is shown to be closely paralleled by (and hence correlated with) ATP synthesis. Acidification of the medium in the presence of valinomycin, under conditions of low external potassium, brings about ATP synthesis in the dark.

Few-cycle solitons in the medium with permanent dipole moment under conditions of the induced birefringence

NASA Astrophysics Data System (ADS)

Sazonov, S. V.

2016-12-01

Propagation of electromagnetic pulse in the birefringent medium consisting of symmetric and asymmetrical molecules is investigated. Stationary quantum states of asymmetrical molecules have the permanent dipole moment. Under considered conditions the ordinary pulse component excites quantum transitions between stationary states. The extraordinary component, besides, causes a dynamic chirp of frequencies of these transitions. The new solitonic modes of propagation of the half- and single-period pulses are found. The solitonic mechanism of simultaneous generation of the second and zero harmonics in the modes of "bright" and "dark" solitons is analyzed.

Production of Normal Mammalian Organ Culture Using a Medium Containing Mem-Alpha, Leibovitz L 15, Glucose Galactose Fructose

NASA Technical Reports Server (NTRS)

Goodwin, Thomas J. (Inventor); Wolf, David A. (Inventor); Spaulding, Glenn F. (Inventor); Prewett, Tacey L. (Inventor)

1999-01-01

Normal mammalian tissue and the culturing process has been developed for the three groups of organ, structural and blood tissue. The cells are grown in vitro under micro- gravity culture conditions and form three dimensional cells aggregates with normal cell function. The microgravity culture conditions may be microgravity or simulated microgravity created in a horizontal rotating wall culture vessel. The medium used for culturing the cells, especially a mixture of epithelial and mesenchymal cells contains a mixture of Mem-alpha and Leibovits L15 supplemented with glucose, galactose and fructose.

Effects of age and brightness contrast on perception of the Wundt-Hering illusion.

PubMed

Astor-Stetson, E; Purnell, T G

1990-10-01

Susceptibility to the Wundt-Hering illusion was studied as a function of age and contrast. Preschoolers, third-graders and college students were shown light-grey, medium-grey, and black Wundt-Hering figures on white ground. Pre-schoolers were most susceptible to the illusion, differing from third graders in the medium and high contrast conditions and from college students in all contrast conditions. Low contrast figures resulted in significantly less distortion than did high contrast figures for the preschoolers. The significant interaction of age and contrast effects highlights the importance of a developmental approach to the study of illusions.

Repetition Blindness for Natural Images of Objects with Viewpoint Changes

PubMed Central

Buffat, Stéphane; Plantier, Justin; Roumes, Corinne; Lorenceau, Jean

2013-01-01

When stimuli are repeated in a rapid serial visual presentation (RSVP), observers sometimes fail to report the second occurrence of a target. This phenomenon is referred to as “repetition blindness” (RB). We report an RSVP experiment with photographs in which we manipulated object viewpoints between the first and second occurrences of a target (0°, 45°, or 90° changes), and spatial frequency (SF) content. Natural images were spatially filtered to produce low, medium, or high SF stimuli. RB was observed for all filtering conditions. Surprisingly, for full-spectrum (FS) images, RB increased significantly as the viewpoint reached 90°. For filtered images, a similar pattern of results was found for all conditions except for medium SF stimuli. These findings suggest that object recognition in RSVP are subtended by viewpoint-specific representations for all spatial frequencies except medium ones. PMID:23346069

l-Asparaginase from Proteus vulgaris1

PubMed Central

Tosa, Tetsuya; Sano, Ryujiro; Yamamoto, Kozo; Nakamura, Masatoshi; Ando, Katsuko; Chibata, Ichiro

1971-01-01

To produce an immunologically and enzymologically new type of l-asparaginase, 108 strains of bacteria were screened for enzyme production. As a result, 13 bacteria belonging to the genera Alcaligenes, Bacterium, and Proteus were found to produce l-asparaginases in high levels. Among these l-asparaginases, partially purified l-asparaginases from B. cadaveris and P. vulgaris showed antitumor activity. A partially purified l-asparaginase preparation of P. vulgaris did not react with the antibody of Escherichia colil-asparaginase on the Ouchterlony agar plate. Culture conditions for the production of l-asparaginase by P. vulgaris were investigated in detail. The enzyme was produced in high yields when cells were grown aerobically in a medium containing sodium fumarate and corn steep liquor. The addition of glucose or ammonium ion to the medium, however, resulted in depressed production of l-asparaginase. Under the optimum conditions, 3,700 international units of l-asparaginase was obtained from 1 liter of culture medium. Images PMID:5000866

Energy and mass balance in the three-phase interstellar medium

NASA Technical Reports Server (NTRS)

Wang, Zhong; Cowie, Lennox L.

1988-01-01

Details of the energy and mass balances are considered in the context of a three-phase interstellar medium. The rates of mass exchange between the different phases are derived based on the pressure variations created by supernova remnant expansions. It is shown that the pressure-confined warm and cold gases have stable temperatures under a variety of interstellar conditions. The three-phase quasi-static configuration is found to be a natural outcome, and both warm and cold phases generally contribute about half of the total mass density to the diffuse interstellar gas. The model is also likely to be self-regulatory in the sense that variations of the input parameters do not strongly alter the general result, which is consistent with most current observations. The consequences of extreme conditions on this model are considered, and the possible implications for interstellar medium in other galaxies are briefly discussed.

Quantum correlation of fiber-based telecom-band photon pairs through standard loss and random media.

PubMed

Sua, Yong Meng; Malowicki, John; Lee, Kim Fook

2014-08-15

We study quantum correlation and interference of fiber-based telecom-band photon pairs with one photon of the pair experiencing multiple scattering in a random medium. We measure joint probability of two-photon detection for signal photon in a normal channel and idler photon in a channel, which is subjected to two independent conditions: standard loss (neutral density filter) and random media. We observe that both conditions degrade the correlation of signal and idler photons, and depolarization of the idler photon in random medium can enhance two-photon interference at certain relative polarization angles. Our theoretical calculation on two-photon polarization correlation and interference as a function of mean free path is in agreement with our experiment data. We conclude that quantum correlation of a polarization-entangled photon pair is better preserved than a polarization-correlated photon pair as one photon of the pair scatters through a random medium.

Ultrastructure of cuticle deposited inPlodia interpunctella wing discs after variousβ-ecdysone treatments in vitro.

PubMed

Dutkowski, A B; Oberlander, H; Leach, C E

1977-06-01

Wing discs of the Indian meal moth may be cultured for extended periods in vitro. The discs produced a tanned cuticle after continuous incubation with β-ecdysone in medium conditioned with fat body or after a 24-h pulse incubation with β-ecdysone in plain medium. We investigated the ultrastructure of the cuticle deposited by such discs. We found that the treatment that produced the most complete cuticle in vitro was the 24-h pulse of hormone. We observed that cuticle formation in vitro was not "all-or-none." Depending on culture conditions, discs produced cuticulin only, complete epicuticle, epicuticle plus diffuse endocuticle, epicuticle plus lamellate endocuticle, or even multiple layers of cuticle. The ultrastructural evidence suggests that continuous incubation with β-ecdysone in plain medium does not always inhibit cuticle formationper se, but does prevent tanning of the partially formed cuticle.

Nutrient requirements and other factors involved in the culture of human kidney cells on microcarrier beads

NASA Technical Reports Server (NTRS)

Lewis, Marian L.; Morrison, Dennis R.

1987-01-01

The culture of human kidney cells on microcarrier beads in the Bioprocessing Laboratory at the Johnson Space Center is described. These were the first series of studies performed before and during 1983 to determine optimum conditions, including medium type, bead type and density. The composition of several medium types and the molecular weights of some common culture medium supplements and cellular proteins are included. The microgravity cell-to-bead attachment experiment performed on Space Transportation System Flight 8 is described.

The Computer as a Medium for Art: Aesthetics and the Processes of Mind.

ERIC Educational Resources Information Center

Woodard, William Bryan

This master's thesis is intended to show that the computer, as an aesthetic medium, will create the social conditions whereby the population will be able to develop the aesthetic bridge of self-actualization, and that the increased numbers of people involved with their own personal psychological development and growth will create social conditions…

Mercury Methylation by Interspecies Hydrogen and Acetate Transfer between Sulfidogens and Methanogens†

PubMed Central

Pak, K.-R.; Bartha, R.

1998-01-01

Cocultures of Desulfovibrio desulfuricans and Methanococcus maripaludis grew on sulfate-free lactate medium while vigorously methylating Hg2+. Individually, neither bacterium could grow or methylate mercury in this medium. Similar synergistic growth of sulfidogens and methanogens may create favorable conditions for Hg2+ methylation in low-sulfate anoxic freshwater sediments. PMID:9603804

[Importance of occupational medicine problem on small and medium scale business enterprises].

PubMed

Fasikov, R M; Stepanov, E G

2008-01-01

First stage of hygienic and psychosocial studies on evaluating the conditions, work specification and health state of small and medium scale business enterprise workers is finished. Findings are that psychosocial factors of work process are highly significant for those workers, and locomotor disorders (backaches) are leaders in morbidity structure among those workers.

Dissociation of heavy quarkonia in an anisotropic hot QCD medium in a quasiparticle model

NASA Astrophysics Data System (ADS)

Jamal, Mohammad Yousuf; Nilima, Indrani; Chandra, Vinod; Agotiya, Vineet Kumar

2018-05-01

The present article is the follow-up work of Phys. Rev. D 94, 094006 (2016), 10.1103/PhysRevD.94.094006, where we have extended the study of quarkonia dissociation in (momentum) anisotropic hot QCD medium. As evident by the experimentally observed collective flow at the RHIC and LHC, the momentum anisotropy is present at almost all the stages after the collision, and therefore, it is important to include its effects in the analysis. Employing the in-medium (corrected) potential while considering the anisotropy (both oblate and prolate cases) in the medium, the thermal widths and the binding energies of the heavy quarkonia states (s -wave charmonia and s -wave bottomonia specifically, for radial quantum numbers n =1 and 2) have been determined. The hot QCD medium effects have been included by employing a quasiparticle description. The presence of anisotropy has modified the potential and then the thermal widths and binding energies of these states in a significant manner. The results show a quite visible shift in the values of dissociation temperatures as compared to the isotropic case. Further, the hot QCD medium interaction effects suppress the dissociation temperature as compared to the case where we consider the medium as a noninteracting ultrarelativistic gas of quarks (antiquarks) and gluons.

Optical imaging of objects in turbid medium with ultrashort pulses

NASA Astrophysics Data System (ADS)

Wang, Chih-Yu; Sun, Chia-Wei; Yang, Chih Chung; Kiang, Yean-Woei; Lin, Chii-Wann

2000-07-01

Photons are seriously scattered when entering turbid medium; this the images of objects hidden in turbid medium can not be obtained by just collecting the transmitted photons. Early-arriving photons, which are also called ballistic or snake protons, are much less scattered when passing through turbid medium, and contains more image information than the late-arriving ones. Therefore, objects embedded in turbid medium can be imaged by gathering the ballistic and snake photons. In the present research we try to recover images of objects in turbid medium by simultaneously time-gate and polarization-gate to obtain the snake photons. An Argon-pumped Ti-Sapphire laser with 100fs pulses was employed as a light source. A streak camera with a 2ps temporal resolution was used to extract the ballistic and snake photons. Two pieces of lean swine meat, measured 4mmX3mm and 5xxX4mm, respectively, were placed in a 10cmX10cmX3cm acrylic tank, which was full of diluted milk. A pair of polarizer and an analyzer was used to extract the light that keeps polarization unchanged. The combination of time gating and polarization gating resulted in good images of objects hidden in turbid medium.

Unified multiphase modeling for evolving, acoustically coupled systems consisting of acoustic, elastic, poroelastic media and septa

NASA Astrophysics Data System (ADS)

Lee, Joong Seok; Kang, Yeon June; Kim, Yoon Young

2012-12-01

This paper presents a new modeling technique that can represent acoustically coupled systems in a unified manner. The proposed unified multiphase (UMP) modeling technique uses Biot's equations that are originally derived for poroelastic media to represent not only poroelastic media but also non-poroelastic ones ranging from acoustic and elastic media to septa. To recover the original vibro-acoustic behaviors of non-poroelastic media, material parameters of a base poroelastic medium are adjusted depending on the target media. The real virtue of this UMP technique is that interface coupling conditions between any media can be automatically satisfied, so no medium-dependent interface condition needs to be imposed explicitly. Thereby, the proposed technique can effectively model any acoustically coupled system having locally varying medium phases and evolving interfaces. A typical situation can occur in an iterative design process. Because the proposed UMP modeling technique needs theoretical justifications for further development, this work is mainly focused on how the technique recovers the governing equations of non-poroelastic media and expresses their interface conditions. We also address how to describe various boundary conditions of the media in the technique. Some numerical studies are carried out to demonstrate the validity of the proposed modeling technique.

Nucleon properties in the Polyakov quark-meson model

NASA Astrophysics Data System (ADS)

Li, Yingying; Hu, Jinniu; Mao, Hong

2018-05-01

We study the nucleon as a nontopological soliton in a quark medium as well as in a nucleon medium in terms of the Polyakov quark-meson (PQM) model with two flavors at finite temperature and density. The constituent quark masses evolving with the temperature at various baryon chemical potentials are calculated and the equations of motion are solved according to the proper boundary conditions. The PQM model predicts an increasing size of the nucleon and a reduction of the nucleon mass in both hot environment. However, the phase structure is different from each other in quark and nucleon mediums. There is a crossover in the low-density region and a first-order phase transition in the high-density region in quark medium, whereas there exists a crossover characterized by the overlap of the nucleons in nucleon medium.

Comparison of different incubation conditions for microbiological environmental monitoring.

PubMed

Gordon, Oliver; Berchtold, Manfred; Staerk, Alexandra; Roesti, David

2014-01-01

Environmental monitoring represents an integral part of the microbiological quality control system of a pharmaceutical manufacturing operation. However, guidance documents differ regarding recommendation of a procedure, particularly regarding incubation time, incubation temperature, or nutrient media. Because of these discrepancies, many manufacturers decide for a particular environmental monitoring sample incubation strategy and support this decision with validation data. Such validations are typically laboratory-based in vitro studies, meaning that these are based on comparing incubation conditions and nutrient media through use of cultured microorganisms. An informal survey of the results of these in vitro studies performed at Novartis or European manufacturing sites of different pharmaceutical companies highlighted that no consensus regarding the optimal incubation conditions for microbial recovery existed. To address this question differently, we collected a significant amount of samples directly from air, inanimate surfaces, and personnel in pharmaceutical production and packaging rooms during manufacturing operation (in situ study). Samples were incubated under different conditions suggested in regulatory guidelines, and recovery of total aerobic microorganisms as well as moulds was assessed. We found the highest recovery of total aerobic count from areas with personnel flow using a general microbiological growth medium incubated at 30-35 °C. The highest recovery of moulds was obtained with mycological medium incubated at 20-25 °C. Single-plate strategies (two-temperature incubation or an intermediate incubation temperature of 25-30 °C) also yielded reasonable recovery of total aerobic count and moulds. However, recovery of moulds was found to be highly inefficient at 30-35 °C compared to lower incubation temperatures. This deficiency could not be rectified by subsequent incubation at 20-25 °C. A laboratory-based in vitro study performed in parallel was inconclusive. We consider our results potentially conferrable to other pharmaceutical manufacturing sites in moderate climate zones and believe that these should represent a valuable reference for definition of the incubation strategy of microbiological environmental monitoring samples. Microbiological environmental monitoring confirms that pharmaceutical cleanrooms are in an appropriate hygienic condition for manufacturing of drug products. Guidance documents from different health authorities or expert groups differ regarding recommendation of the applied incubation time, incubation temperature, or nutrient media. Therefore, many pharmaceutical manufacturers perform studies that aim to identify the optimal incubation setup for environmental monitoring samples. An informal survey of the results of such studies, which had been performed at Novartis or European manufacturing sites of different pharmaceutical companies, highlighted no consensus regarding the optimal incubation conditions for microbial recovery. All these studies had been conducted in the laboratory using selections of cultured microbial strains. We tried to solve this disagreement by collecting a significant amount of real environmental monitoring samples directly from the environment in pharmaceutical production and packaging rooms during manufacturing operation. These samples were then incubated under different conditions suggested in the regulatory guidelines. We believe that the results of our study are more meaningful than laboratory-based experiments because we used environmental samples with microorganisms directly isolated from the manufacturing area. Therefore, we believe that our results should represent a valuable reference for definition of the incubation strategy of microbiological environmental monitoring samples. © PDA, Inc. 2014.

Shaft seal system

DOEpatents

Kapich, Davorin D.

1985-01-01

A shaft seal system is disclosed for isolating two regions of different fluid mediums through which a rotatable shaft extends. The seal system includes a seal housing through which the shaft extends and which defines an annular land and an annular labyrinth both of which face on the shaft so that each establishes a corresponding fluid sealing annulus. A collection cavity is formed in communication with the annular sealing spaces, and fluids compatible with the fluids in each of the two regions to be isolated are introduced, respectively, into the annular sealing spaces and collected in the collection cavity from which the fluid mixture is removed and passed to a separator which separates the fluids and returns them to their respective annular sealing spaces in a recycling manner. In the illustrated embodiment, the isolated fluid mediums comprise a liquid region and a gas region. Gas is removed from the gas region and passed through a purifier and a gas pump operative to introduce the purified gas through the labyrinth sealing annulus to the collection cavity. After passing to the separator, the separated gas is passed through a dryer from which the dried gas is caused to pass through the labyrinth sealing annulus into the collection cavity independently of the purified gas so as to insure isolation of the gas region in the event of sealing gas pump malfunction.

Large Fizeau's light-dragging effect in a moving electromagnetically induced transparent medium.

PubMed

Kuan, Pei-Chen; Huang, Chang; Chan, Wei Sheng; Kosen, Sandoko; Lan, Shau-Yu

2016-10-03

As one of the most influential experiments on the development of modern macroscopic theory from Newtonian mechanics to Einstein's special theory of relativity, the phenomenon of light dragging in a moving medium has been discussed and observed extensively in different types of systems. To have a significant dragging effect, the long duration of light travelling in the medium is preferred. Here we demonstrate a light-dragging experiment in an electromagnetically induced transparent cold atomic ensemble and enhance the dragging effect by at least three orders of magnitude compared with the previous experiments. With a large enhancement of the dragging effect, we realize an atom-based velocimeter that has a sensitivity two orders of magnitude higher than the velocity width of the atomic medium used. Such a demonstration could pave the way for motional sensing using the collective state of atoms in a room temperature vapour cell or solid state material.

Evaluation of RGM Medium for Isolation of Nontuberculous Mycobacteria from Respiratory Samples from Patients with Cystic Fibrosis in the United States.

PubMed

Plongla, Rongpong; Preece, Clair L; Perry, John D; Gilligan, Peter H

2017-05-01

A novel selective agar (RGM medium) has been advocated for the isolation of rapidly growing mycobacteria from the sputa of cystic fibrosis (CF) patients. The aim of this study was to compare RGM medium to Burkholderia cepacia selective agar (BCSA) and a standard acid-fast bacillus (AFB) culture method for the isolation of nontuberculous mycobacteria (NTM) from patients with CF. The applicability of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) for the identification of NTM isolated on RGM medium was also assessed. Respiratory samples ( n = 869) were collected from 487 CF patients and inoculated directly onto RGM medium and BCSA. Cultures were incubated at 30°C and examined for up to 28 days. A subset of 212 samples (from 172 patients) was also cultured by using a mycobacterial growth indicator tube (MGIT) and on Lowenstein-Jensen medium following dual decontamination. By using a combination of all methods, 98 mycobacteria were isolated from 869 samples (11.3%). The sensitivity of RGM medium (96.9%) was significantly higher than that of BCSA (35.7%) for the isolation of mycobacteria ( P < 0.0001). The sensitivity of RGM medium was also superior to that of standard AFB culture for the isolation of mycobacteria (92.2% versus 47.1%; P < 0.0001). MALDI-TOF MS was effective for the identification of mycobacteria in RGM medium. RGM medium offers a simple and highly effective tool for the isolation of NTM from patients with CF. Extended incubation of RGM medium for 28 days facilitates the isolation of slow-growing species, including members of the Mycobacterium avium complex (MAVC). Copyright © 2017 American Society for Microbiology.

Influence of natural organic matter in porous media on fine particle transport.

PubMed

Zhou, Yuhong; Cheng, Tao

2018-06-15

Although extensive research has been conducted to understand the effects of dissolved organic matter (DOM) on fine particle transport, less attention has been paid to natural organic matter (NOM) in the transport medium (i.e., immobile rock and sediment grains). The objective of this study is to elucidate the roles of NOM in the transport medium in mediating particle transport. We conducted experimental and modelling study on the transport of nanoscale titanium dioxide (nTiO 2 ) and illite colloid in columns packed with quartz sand under water-saturated conditions. Peat moss was used as an example NOM and packed in some of the columns to investigate its influence on particle transport. Experimental results showed that NOM may either increase or decrease particle transport depending on the specific conditions. NOM in the transport medium was found to attract particles and reduce particle mobility when the energy barrier between particle and NOM is low or non-existent. NOM also adsorb to Fe and Al oxyhydroxides and promote the transport of negatively-charged particles at low pH. Partial dissolution of NOM releases DOM, and the DOM adsorbs to and increases the transport of positively-charged particles. Additionally, NOM changes pore water pH, which influences particle mobility by affecting the interaction energy between the particle and transport medium. Modelling results showed that the deposition sites provided by peat moss are very heterogeneous, and the NOM from peat moss may reduce particle deposition rate by adsorbing to the particle and/or transport medium. Findings from this study demonstrate that NOM in the transport medium not only changes property of the medium, but also may alter water chemistry. Therefore, the role of NOM in mediating particle transport is complicated and dependent on the property of the particle, NOM, and mineralogical composition of the medium. Copyright © 2018 Elsevier B.V. All rights reserved.

Screening of Actinomycetes from mangrove ecosystem for L-asparaginase activity and optimization by response surface methodology.

PubMed

Usha, Rajamanickam; Mala, Krishnaswami Kanjana; Venil, Chidambaram Kulandaisamy; Palaniswamy, Muthusamy

2011-01-01

Marine actinomycetes were isolated from sediment samples collected from Pitchavaram mangrove ecosystem situated along the southeast coast of India. Maximum actinomycete population was noted in rhizosphere region. About 38% of the isolates produced L-asparaginase. One potential strain KUA106 produced higher level of enzyme using tryptone glucose yeast extract medium. Based on the studied phenotypic characteristics, strain KUA106 was identified as Streptomyces parvulus KUA106. The optimization method that combines the Plackett-Burman design, a factorial design and the response surface method, which were used to optimize the medium for the production of L-asparaginase by Streptomycetes parvulus. Four medium factors were screened from eleven medium factors by Plackett-Burman design experiments and subsequent optimization process to find out the optimum values of the selected parameters using central composite design was performed. Asparagine, tryptone, d) extrose and NaCl components were found to be the best medium for the L-asparaginase production. The combined optimization method described here is the effective method for screening medium factors as well as determining their optimum level for the production of L-asparaginase by Streptomycetes parvulus KUAP106.

A selective medium for the isolation of Microbacterium species in oral cavities.

PubMed

Tsuzukibashi, Osamu; Uchibori, Satoshi; Kobayashi, Taira; Saito, Masanori; Umezawa, Koji; Ohta, Mitsuhiro; Shinozaki-Kuwahara, Noriko

2015-09-01

The genus Microbacterium has been isolated from the environment, dairy goods, and human clinical specimens. Although, in our previous studies, some Microbacterium species were infrequently detected in oral samples collected from humans, there is currently no report that these organisms, which are capable of causing serious systemic infections, were isolated from the human oral cavity. The aim of the present study was to develop a selective medium to isolate the representative Microbacterium species most frequently detected in human clinical specimens, and reveal the distribution of individual Microbacterium species in the oral cavity. The growth recoveries of representative Microbacterium species on the selective medium, designated as MSM, were sufficient. Moreover, the growth of other representative oral bacteria was markedly inhibited on the selective medium. The proportion of Microbacterium species in the saliva samples of 60 subjects, 20 of whom were removable denture wearers, was then examined. The proportion of these organisms was also examined in environmental samples obtained by swabbing 20 washstands. PCR primers were designed for representative Microbacterium species. The genus Microbacterium was detected in 45% of the saliva and denture plaque samples collected from the twenty removable denture wearers, but was absent in the saliva of the forty non-denture wearers. On the other hand, these organisms were detected in all environmental samples. The genus Microbacterium accounted for 0.00003%, 0.0001%, and 12.6% of the total cultivable bacteria number on the BHI medium in the saliva and denture plaque samples of removable denture wearers and in the environmental samples, respectively. The most predominant Microbacterium species in all positive samples was Microbacterium oxydans. These results indicated that the genus Microbacterium was not a part of the normal flora in the human oral cavity, except for subjects wearing dentures that were contaminated by the environment, and the selective medium, designated as MSM, was useful for isolating Microbacterium species, which are frequently encountered in human clinical specimens, from the various samples. Copyright © 2015 Elsevier B.V. All rights reserved.

TGF-β signaling is required for maintenance of retinal ganglion cell differentiation and survival.

PubMed

Walshe, T E; Leach, L L; D'Amore, P A

2011-08-25

To determine the role of TGF-β1 in the maintenance of retinal ganglion cell line (RGC-5) differentiation and integrity. RGC-5 cells were differentiated in media conditioned by human non-pigmented ciliary epithelial cells (HNPE) for 4 days before treatment with TGF-β1 for 24 h. Cells were examined for morphological changes and harvested for western blot and real-time PCR analysis. For study of apoptosis, differentiated RGC-5 cells were grown in serum-free medium for 24 h in the presence or absence of TGF-β1 and collected for Annexin V/Propidium iodide FACs analysis. The role of MAPK pathways in TGF-β1-dependent signaling was determined by treatment with specific inhibitors of ERK, JNK and p38. Differentiation of RGC-5 cells in HNPE-conditioned media (CM) increased the neural cell markers, Brn-3c, NF-160, Thy1.2, Tau and PGP9.5. Treatment with TGF-β1 significantly increased the length of neurites extended by differentiated RGC-5s, concomitant with increased expression of NF-160 and PGP9.5, but not Brn-3c, Thy1.2 or Tau. TGF-β1 also decreased RGC-5 cell apoptosis in serum-free medium. p38 phosphorylation, but not smad2/3, JNK or ERK phosphorylation, was increased in TGF-β1 treated cells. Specific inhibition of p38 signaling reversed TGF-β1 induced neurite growth. These findings demonstrate the induction of RGC-5 cell differentiation by HNPE-derived CM and illustrate a role for TGF-β1 in maintaining RGC-5 cell survival and promoting neurite outgrowth through p38 MAPK. Copyright © 2011 IBRO. Published by Elsevier Ltd. All rights reserved.

Foodborne pathogens in unpasteurized milk in Sweden.

PubMed

Artursson, Karin; Schelin, Jenny; Thisted Lambertz, Susanne; Hansson, Ingrid; Olsson Engvall, Eva

2018-05-19

Raw milk may be a risk for public health if it is contaminated with zoonotic pathogens. To study the prevalence in unpasteurized milk from Swedish farms, bovine and small ruminant dairy farms were sampled. Since the sampling method and transport conditions may influence the outcome of analyses, efforts were made to optimize the methodology. Culturing of bacteria was done from in-line milk filters collected from the milk pipe at the point where it enters the milk bulk tank at the farms and this way of sampling was compared to sampling bulk tank milk (BTM) directly. Analysing milk filters were found to be superior to analysing BTM directly. Conditions for transport of milk filter samples were further improved by the addition of Cary Blair transport medium, which significantly increased the number of positive samples for pathogenic bacteria. The isolation of several foodborne pathogens from milk filters was demonstrated. The prevalence of samples with Staphylococcus aureus was 71% and 64%, and Listeria spp. 21% and 29% from dairy cow and goat/sheep farms, respectively. Campylobacter jejuni, Yersinia enterocolitica and verotoxigenic Escherichia coli (VTEC) O157 were detected in 9%, 2% and 2% of samples from bovine milk, respectively. We conclude that the choice of sampling method and sample handling influence the results of bacterial culturing. From the results of this study, we strongly recommend to sample in-line milk filters instead of BTM directly and to use Cary Blair medium during transport, especially if the samples are to be analysed for Campylobacter spp. and/or Listeria spp. The findings also show that unpasteurized milk from Swedish farms occasionally contain bacteria with zoonotic potential. Copyright © 2018 Elsevier B.V. All rights reserved.

The influence of serum substituents on serum-free Vero cell conditioned culture media manufactured from Dulbecco's modified Eagle medium in mouse embryo culture.

PubMed

Lee, Jong-Seon; Kim, Ju-Hwan; Seo, Young-Seok; Yang, Jung-Bo; Kim, Yong-Il; Kim, Hye-Jin; Lee, Ki-Hwan

2013-09-01

This study was conducted to examine the influences of supplementation of the serum substituents and available period of serum-free Vero cell conditioned media (SF-VCM) manufactured from Dulbecco's modified Eagle medium cultured with Vero cells for in vitro development of mouse preimplantation embryos. A total of 1,099 two-cell embryos collected from imprinting control region mice were cultured in SF-VCM with 10% and 20% human follicular fluid (hFF), serum substitute supplement (SSS), and serum protein substitute (SPS). Development of embryos was observed every 24 hours. Results between different groups were analyzed by chi-square test, and considered statistically significant when P-value was less than 0.05. The rates of embryonic development cultured in SF-VCM supplemented with serum substituents were significantly higher compare with serum-free group (P < 0.05). The rates of embryonic development after 48 hours (morula≤) and 96 hours (blastocyst≤) were significantly higher in 20% SSS and 10% SPS than in 20% hFF supplementation (P < 0.05). And the rates of embryonic development after 96 hours (hatching blastocyst≤) were significantly higher in 10% SPS (94.5%) than in 20% SSS (82.6%) and 20% hFF supplementation (68.5%). The rates of embryonic development according to storage period of the SF-VCM supplemented with 10% SPS showed no significant difference between control, 2 weeks and 4 weeks group. However developmental rate in 6 weeks storage group was significantly lower than other groups. The rate of embryonic development after 96 hours (hatching blastocyst≤) was significantly higher in SF-VCM supplemented with 10% SPS. And storage period of media up to 4 weeks did not affect on embryonic development.

Conditioned Medium from Periodontal Ligament Stem Cells Enhances Periodontal Regeneration.

PubMed

Nagata, Mizuki; Iwasaki, Kengo; Akazawa, Keiko; Komaki, Motohiro; Yokoyama, Naoki; Izumi, Yuichi; Morita, Ikuo

2017-05-01

Periodontal disease is one of the most common infectious diseases in adults and is characterized by the destruction of tooth-supporting tissues. Mesenchymal stem cells (MSCs) comprise the mesoderm-originating stem cell population, which has been studied and used for cell therapy. However, because of the lower rate of cell survival after MSC transplantation in various disease models, paracrine functions of MSCs have been receiving increased attention as a regenerative mechanism. The aim of this study was to investigate the regenerative potential of transplanted conditioned medium (CM) obtained from cultured periodontal ligament stem cells (PDLSCs), the adult stem cell population in tooth-supporting tissues, using a rat periodontal defect model. Cell-free CM was collected from PDLSCs and fibroblasts, using ultrafiltration and transplanted into surgically created periodontal defects. Protein content of CM was examined by antibody arrays. Formation of new periodontal tissues was analyzed using microcomputed tomography and histological sections. PDLSC-CM transplantation enhanced periodontal tissue regeneration in a concentration-dependent manner, whereas fibroblast-CM did not show any regenerative function. Proteomic analysis revealed that extracellular matrix proteins, enzymes, angiogenic factors, growth factors and cytokines were contained in PDLSC-CM. Furthermore, PDLSC-CM transplantation resulted in the decreased mRNA level of tumor necrosis factor-α (TNF-α) in healing periodontal tissues. In addition, we found that PDLSC-CM suppressed the mRNA level of TNF-α in the monocyte/macrophage cell line, RAW cells, stimulated with IFN-γ. Our findings suggested that PDLSC-CM enhanced periodontal regeneration by suppressing the inflammatory response through TNF-α production, and transplantation of PDLSC-CM could be a novel approach for periodontal regenerative therapy.

Conditioned Medium from Periodontal Ligament Stem Cells Enhances Periodontal Regeneration

PubMed Central

Nagata, Mizuki; Akazawa, Keiko; Komaki, Motohiro; Yokoyama, Naoki; Izumi, Yuichi; Morita, Ikuo

2017-01-01

Periodontal disease is one of the most common infectious diseases in adults and is characterized by the destruction of tooth-supporting tissues. Mesenchymal stem cells (MSCs) comprise the mesoderm-originating stem cell population, which has been studied and used for cell therapy. However, because of the lower rate of cell survival after MSC transplantation in various disease models, paracrine functions of MSCs have been receiving increased attention as a regenerative mechanism. The aim of this study was to investigate the regenerative potential of transplanted conditioned medium (CM) obtained from cultured periodontal ligament stem cells (PDLSCs), the adult stem cell population in tooth-supporting tissues, using a rat periodontal defect model. Cell-free CM was collected from PDLSCs and fibroblasts, using ultrafiltration and transplanted into surgically created periodontal defects. Protein content of CM was examined by antibody arrays. Formation of new periodontal tissues was analyzed using microcomputed tomography and histological sections. PDLSC-CM transplantation enhanced periodontal tissue regeneration in a concentration-dependent manner, whereas fibroblast-CM did not show any regenerative function. Proteomic analysis revealed that extracellular matrix proteins, enzymes, angiogenic factors, growth factors and cytokines were contained in PDLSC-CM. Furthermore, PDLSC-CM transplantation resulted in the decreased mRNA level of tumor necrosis factor-α (TNF-α) in healing periodontal tissues. In addition, we found that PDLSC-CM suppressed the mRNA level of TNF-α in the monocyte/macrophage cell line, RAW cells, stimulated with IFN-γ. Our findings suggested that PDLSC-CM enhanced periodontal regeneration by suppressing the inflammatory response through TNF-α production, and transplantation of PDLSC-CM could be a novel approach for periodontal regenerative therapy. PMID:28027709

Evaluating ambulatory care training in Firoozgar hospital based on Iranian national standards of undergraduate medical education

PubMed Central

Sabzghabaei, Foroogh; Salajeghe, Mahla; Soltani Arabshahi, Seyed Kamran

2017-01-01

Background: In this study, ambulatory care training in Firoozgar hospital was evaluated based on Iranian national standards of undergraduate medical education related to ambulatory education using Baldrige Excellence Model. Moreover, some suggestions were offered to promote education quality in the current condition of ambulatory education in Firoozgar hospital and national standards using the gap analysis method. Methods: This descriptive analytic study was a kind of evaluation research performed using the standard check lists published by the office of undergraduate medical education council. Data were collected through surveying documents, interviewing, and observing the processes based on the Baldrige Excellence Model. After confirming the validity and reliability of the check lists, we evaluated the establishment level of the national standards of undergraduate medical education in the clinics of this hospital in the 4 following domains: educational program, evaluation, training and research resources, and faculty members. Data were analyzed according to the national standards of undergraduate medical education related to ambulatory education and the Baldrige table for scoring. Finally, the quality level of the current condition was determined as very appropriate, appropriate, medium, weak, and very weak. Results: In domains of educational program 62%, in evaluation 48%, in training and research resources 46%, in faculty members 68%, and in overall ratio, 56% of the standards were appropriate. Conclusion: The most successful domains were educational program and faculty members, but evaluation and training and research resources domains had a medium performance. Some domains and indicators were determined as weak and their quality needed to be improved, so it is suggested to provide the necessary facilities and improvements by attending to the quality level of the national standards of ambulatory education PMID:29951400

Tooth development in vitro in two teleost fish, the cichlid Hemichromis bimaculatus and the cyprinid Danio rerio.

PubMed

Van der Heyden, C; Allizard, F; Sire, J-Y; Huysseune, A

2005-09-01

A technique for organotypic in vitro culture with serum-free medium was tested for its appropriateness to mimic normal odontogenesis in the cichlid fish Hemichromis bimaculatus and the zebrafish Danio rerio. Serial semithin sections were observed by light microscopy to collect data on tooth patterning and transmission electron microscopy was used to compare cellular and extracellular features of tooth germs developing in vitro with the situation in vivo. Head explants of H. bimaculatus from 120 h post-fertilization (hPF) to 8.5 days post-fertilization (dPF) and of zebrafish from 45 hPF to 79 hPF and adults kept in culture for 3, 4 or 7 days revealed that tooth germs developed in vitro from explants in which the buccal or pharyngeal epithelium was apparently undifferentiated and, when present at the time of explantation, they continued their development up to a stage of attachment. In addition, the medium allowed the morphogenesis and cytodifferentiation of the tooth germs similar to that observed in vivo and the establishment of a dental pattern (place and order of tooth appearance and of attachment) that mimicked that in vivo. Organotypic culture in serum-free conditions thus provides us with the means of studying epithelial-mesenchymal interactions during tooth development in teleost fish and of analysing the genetic control of either mandibular or pharyngeal tooth development and replacement in these polyphyodont species. Importantly, it allows heads from embryonically lethal (zebrafish) mutants or from early lethal knockdown experiments to develop beyond the point at which the embryos normally die. Such organotypic culture in serum-free conditions could therefore become a powerful tool in developmental studies and open new perspectives for craniofacial research.

Guidelines for evaluating fish habitat in Wisconsin streams.

Treesearch

Timothy D. Simonson; John Lyons; Paul D. Kanehl

1993-01-01

Describes procedures for evaluating the quality and quantity of habitat for fish in small and medium streams of Wisconsin. Provides detailed guidelines for collecting and analyzing specific quantitative habitat information.

A flow paradigm in heavy-ion collisions

NASA Astrophysics Data System (ADS)

Yan, Li

2018-04-01

The success of hydrodynamics in high energy heavy-ion collisions leads to a flow paradigm, to understand the observed features of harmonic flow in terms of the medium collective expansion with respect to initial state geometrical properties. In this review, we present some essential ingredients in the flow paradigm, including the hydrodynamic modeling, the characterization of initial state geometry and the medium response relations. The extension of the flow paradigm to small colliding systems is also discussed. Supported by Natural Sciences and Engineering Research Council of Canada

Evaluation and selection of bio-relevant dissolution media for a poorly water-soluble new chemical entity.

PubMed

Tang, L; Khan, S U; Muhammad, N A

2001-11-01

The purpose of this work is to develop a bio-relevant dissolution method for formulation screening in order to select an enhanced bioavailable formulation for a poorly water-soluble drug. The methods used included a modified rotating disk apparatus for measuring intrinsic dissolution rate of the new chemical entity (NCE) and the USP dissolution method II for evaluating dissolution profiles of the drug in three different dosage forms. The in vitro dissolution results were compared with the in vivo bioavailability for selecting a bio-relevant medium. The results showed that the solubility of the NCE was proportional to the concentration of sodium lauryl sulfate (SLS) in the media. The apparent intrinsic dissolution rate of the NCE was linear to the rotational speed of the disk, which indicated that the dissolution of the drug is a diffusion-controlled mechanism. The apparent intrinsic dissolution rate was also linear to the surfactant concentration in the media, which was interpreted using the Noyes and Whitney Empirical Theory. Three formulations were studied in three different SLS media using the bulk drug as a reference. The dissolution results were compared with the corresponding bioavailability results in dogs. In the 1% SLS--sink conditions--the drug release from all the formulations was complete and the dissolution results were discriminative for the difference in particle size of the drug in the formulations. However, the data showed poor IVIV correlation. In the 0.5% SLS medium--non-sink conditions--the dissolution results showed the same rank order among the tested formulations as the bioavailability. The best IVIV correlation was obtained from the dissolution in 0.25% SLS medium, an over-saturated condition. The conclusions are: a surfactant medium increases the apparent intrinsic dissolution rate of the NCE linearly due to an increase in solubility. A low concentration of surfactant in the medium (0.25%) is more bio-relevant than higher concentrations of surfactant in the media for the poorly water-soluble drug. Creating sink conditions (based on bulk drug solubilities) by using a high concentration of a surfactant in the dissolution medium may not be a proper approach in developing a bio-relevant dissolution method for a poorly water-soluble drug.

Culturable microbial diversity and the impact of tourism in Kartchner Caverns, Arizona.

PubMed

Ikner, Luisa A; Toomey, Rickard S; Nolan, Ginger; Neilson, Julia W; Pryor, Barry M; Maier, Raina M

2007-01-01

Kartchner Caverns in Benson, AZ, was opened for tourism in 1999 after a careful development protocol that was designed to maintain predevelopment conditions. As a part of an ongoing effort to determine the impact of humans on this limestone cave, samples were collected from cave rock surfaces along the cave trail traveled daily by tour groups (200,000 visitors year-1) and compared to samples taken from areas designated as having medium (30-40 visitors year-1) and low (2-3 visitors year-1) levels of human exposure. Samples were also taken from fiberglass moldings installed during cave development. Culturable bacteria were recovered from these samples and 90 unique isolates were identified by using 16S rRNA polymerase chain reaction and sequencing. Diversity generally decreased as human impact increased leading to the isolation of 32, 27, and 22 strains from the low, medium, and high impact areas, respectively. The degree of human impact was also reflected in the phylogeny of the isolates recovered. Although most isolates fell into one of three phyla: Actinobacteria, Firmicutes, or Proteobacteria, the Proteobacteria were most abundant along the cave trail (77% of the isolates), while Firmicutes predominated in the low (66%) and medium (52%) impact areas. Although the abundance of Proteobacteria along the cave trail seems to include microbes of environmental rather than of anthropogenic origin, it is likely that their presence is a consequence of increased organic matter availability due to lint and other organics brought in by cave visitors. Monitoring of the cave is still in progress to determine whether these bacterial community changes may impact the future development of cave formations.

[Evaluation of soil heavy metals accumulation in the fast economy development region].

PubMed

Zhong, Xian-Lan; Zhou, Sheng-Lu; Li, Jiang-Tao; Zhao, Qi-Guo

2010-06-01

Evaluation of soil heavy metals accumulation was studied in Kunshan City, a typical region of the fast economy development region in China. 126 soil samples were collected and analyzed, and evaluation indexes of soil heavy metal accumulation, which including total concentration of soil heavy metal index (THMI), soil available heavy metal index (AHMI) and fractionation of soil heavy metal index (FHMI), were established, and the heavy metal accumulation conditions of soil in this region were also discussed. Results showed as follows: the spatial variability of THMI was relative lower, with a mean value of 42.57%, whereas strong variability was found in AHMI and FHMI (especially active fraction of soil heavy metals), with the average value of 82.75% and 77.83%, respectively. Judging by each index reference standard of C Horizon, THMI was low-grade with a mean value of 1.01, while the AHMI and FHMI reached to medium accumulation and serious accumulation, with the average values of 2.46 and 4.32, respectively. The synthetic accumulation index of soil heavy metals (SHMI) was 2.56, reaching to medium grade level and with strong variability. 21.54% land area was in low-grade accumulation and 54.70% land area was in medium grade accumulation, while 23.76% land area was in serious accumulation under SHMI evaluation system. All the accumulation evaluation indexes in livestock breeding zone were the lowest, while the indexes in the smelting and plating zone were the highest, but the indexes difference between two zones were unobvious. There were markedly differences in soil types, which the accumulation indexes in Wushan soil were significantly higher than those in Huangni soil and Qingni soil.

Alisol A 24-acetate ameliorates nonalcoholic steatohepatitis by inhibiting oxidative stress and stimulating autophagy through the AMPK/mTOR pathway.

PubMed

Wu, Chenqu; Jing, Menghui; Yang, Lijuan; Jin, Lei; Ding, Yicun; Lu, Juan; Cao, Qin; Jiang, Yuanye

2018-06-05

Alisol A 24-acetate (AA), a natural triterpenoid isolated from the traditional Chinese medicine Rhizoma Alismatis, has various therapeutic effects. We investigated the anti-nonalcoholic steatohepatitis (NASH) effect of AA and its underlying mechanisms in vitro and in vivo. C57BL/6 mice were fed a methionine and choline-deficient (MCD) diet for 4 weeks to induce NASH. The mice were simultaneously treated with a daily dose of AA (15, 30, and 60 mg kg -1 , ig) for 4 weeks. On the last day, the animals were sacrificed and plasma and liver tissue were collected. Serum and liver tissue biochemical analyses and histological observation were performed. The human hepatic stellate cell line LX-2 was used to build NASH models by culturing with conditioned medium from WRL-68 liver cells after exposure to MCD medium in vitro. Liver oxidative stress and inflammatory indices and autophagy markers were examined. The results showed that AA suppressed reactive oxygen species (ROS) and inflammation in a NASH mouse model and inhibited the expression of inflammatory cytokines and ROS in LX-2 cells in MCD medium. Furthermore, we found AA stimulated autophagy in mice liver and LX-2, which could be the underlying mechanism of AA in NASH. To further investigate the role of autophagy in LX-2 cells, we found that AA regulated autophagy via the AMPK/mTOR/ULK1 pathway and dorsomorphin, a selective AMPK inhibitor, led to the suppression of AA-induced autophagy. Taken together, our results indicate that AA could be a possible therapy for NASH by inhibiting oxidative stress and stimulating autophagy. Copyright © 2018. Published by Elsevier B.V.

Assessing the In Vitro Drug Release from Lipid-Core Nanocapsules: a New Strategy Combining Dialysis Sac and a Continuous-Flow System.

PubMed

de Andrade, Diego Fontana; Zuglianello, Carine; Pohlmann, Adriana Raffin; Guterres, Silvia Stanisçuaski; Beck, Ruy Carlos Ruver

2015-12-01

The in vitro assessment of drug release from polymeric nanocapsules suspensions is one of the most studied parameters in the development of drug-loaded nanoparticles. Nevertheless, official methods for the evaluation of drug release from submicrometric carriers are not available. In this work, a new approach to assess the in vitro drug release profile from drug-loaded lipid-core nanocapsules (LNC) was proposed. A continuous-flow system (open system) was designed to evaluate the in vitro drug release profiles from different LNC formulations containing prednisolone or clobetasol propionate (LNC-CP) as drug model (LNC-PD) using a homemade apparatus. The release medium was constantly renewed throughout the experiment. A dialysis bag containing 5 mL of formulation (0.5 mg mL(-1)) was maintained inside the apparatus, under magnetic stirring and controlled temperature (37°C). In parallel, studies based on the conventional dialysis sac technique (closed system) were performed. It was possible to discriminate the in vitro drug release profile of different formulations using the open system. The proposed strategy improved the sink condition, by constantly renewing the release medium, thus maintaining the drug concentration farther from the saturated concentration in the release medium. Moreover, problems due to sampling errors can be easily overcome using this semi-automated system, since the collection is done automatically without interference from the analyst. The system proposed in this paper brings important methodological and analytical advantages, becoming a promising prototype semi-automated apparatus for performing in vitro drug release studies from drug-loaded lipid-core nanocapsules and other related nanoparticle drug delivery systems.

Sperm viability in the black-footed ferret (Mustela nigripes) is influenced by seminal and medium osmolality.

PubMed

Santymire, Rachel M; Marinari, Paul E; Kreeger, Julie S; Wildt, David E; Howard, JoGayle

2006-08-01

Fundamental knowledge of spermatozoa cryobiology can assist with optimizing cryopreservation protocols needed for genetic management of the endangered black-footed ferret. Objectives were to characterize semen osmolality and assess the influence of two media at various osmolalities on sperm viability. We examined the influence of Ham's F10 +Hepes medium (H) at 270, 400, 500 or 700 mOsm (adjusted with sucrose, a nonpermeating cryoprotectant) and TEST Yolk Buffer (TYB) with 0% (300 mOsm) versus 4% (900 mOsm) glycerol (a permeating cryoprotectant). Electroejaculates (n=16) were assessed for osmolality using a vapor pressure osmometer. For media comparison, semen (n=5) was collected in TYB 0%, split into six aliquots, and diluted in H270, H400, H500, H700, and TYB 0% or TYB 4%. Each sample was centrifuged (300 g, 8 min), resuspended in respective medium, and maintained at 37 degrees C for 3h. Sperm motility and forward progression were monitored every 30 min for 3h post-washing. Acrosomal integrity was monitored at 0 and 60 min post-washing. Results demonstrated that black-footed ferret semen has a comparatively high osmolality (mean+/-SEM, 513.1+/-32.6 mOsm; range, 366-791 mOsm). Ferret spermatozoa were sensitive to hyperosmotic stress. Specifically, sperm motility was more susceptible (P<0.01) to hyperosmotic conditions than acrosomal integrity, and neither were influenced (P>0.05) by hypotonic solutions. Exposure to TYB 4% glycerol retained more (P<0.01) sperm motility than a hyperosmotic Ham's (700 mOsm). These findings will guide the eventual development of assisted breeding with cryopreserved sperm contributing to genetic management of this rare species.

World Endometriosis Research Foundation Endometriosis Phenome and Biobanking Harmonisation Project: IV. Tissue collection, processing, and storage in endometriosis research

PubMed Central

Fassbender, Amelie; Rahmioglu, Nilufer; Vitonis, Allison F.; Viganò, Paola; Giudice, Linda C.; D’Hooghe, Thomas M.; Hummelshoj, Lone; Adamson, G. David; Becker, Christian M.; Missmer, Stacey A.; Zondervan, Krina T.; Adamson, G.D.; Allaire, C.; Anchan, R.; Becker, C.M.; Bedaiwy, M.A.; Buck Louis, G.M.; Calhaz-Jorge, C.; Chwalisz, K.; D'Hooghe, T.M.; Fassbender, A.; Faustmann, T.; Fazleabas, A.T.; Flores, I.; Forman, A.; Fraser, I.; Giudice, L.C.; Gotte, M.; Gregersen, P.; Guo, S.-W.; Harada, T.; Hartwell, D.; Horne, A.W.; Hull, M.L.; Hummelshoj, L.; Ibrahim, M.G.; Kiesel, L.; Laufer, M.R.; Machens, K.; Mechsner, S.; Missmer, S.A.; Montgomery, G.W.; Nap, A.; Nyegaard, M.; Osteen, K.G.; Petta, C.A.; Rahmioglu, N.; Renner, S.P.; Riedlinger, J.; Roehrich, S.; Rogers, P.A.; Rombauts, L.; Salumets, A.; Saridogan, E.; Seckin, T.; Stratton, P.; Sharpe-Timms, K.L.; Tworoger, S.; Vigano, P.; Vincent, K.; Vitonis, A.F.; Wienhues-Thelen, U.-H.; Yeung, P.P.; Yong, P.; Zondervan, K.T.

2014-01-01

Objective To harmonize standard operating procedures (SOPs) and standardize the recording of associated data for collection, processing, and storage of human tissues relevant to endometriosis. Design An international collaboration involving 34 clinical/academic centers and three industry collaborators from 16 countries on five continents. Setting In 2013, two workshops were conducted followed by global consultation, bringing together 54 leaders in endometriosis research and sample processing from around the world. Patient(s) None. Intervention(s) Consensus SOPs were based on: 1) systematic comparison of SOPs from 24 global centers collecting tissue samples from women with and without endometriosis on a medium or large scale (publication on >100 cases); 2) literature evidence where available, or consultation with laboratory experts otherwise; and 3) several global consultation rounds. Main Outcome Measure(s) Standard recommended and minimum required SOPs for tissue collection, processing, and storage in endometriosis research. Result(s) We developed “recommended standard” and “minimum required” SOPs for the collection, processing, and storage of ectopic and eutopic endometrium, peritoneum, and myometrium, and a biospecimen data collection form necessary for interpretation of sample-derived results. Conclusion(s) The EPHect SOPs allow endometriosis research centers to decrease variability in tissue-based results, facilitating between-center comparisons and collaborations. The procedures are also relevant to research into other gynecologic conditions involving endometrium, myometrium, and peritoneum. The consensus SOPs are based on the best available evidence; areas with limited evidence are identified as requiring further pilot studies. The SOPs will be reviewed based on investigator feedback and through systematic triannual follow-up. Updated versions will be made available at: http://endometriosisfoundation.org/ephect. PMID:25256928

Research in Brief: Using Mobile Phones to Collect Daily Experience Data from College Undergraduates

ERIC Educational Resources Information Center

Ravert, Russell D.; Calix, Shaun I.; Sullivan, Michael J.

2010-01-01

As mobile phone use and text messaging has continued to play a more central role in people's daily lives, some researchers and clinicians have sought to use the medium as a tool for interventions and data collection. Such efforts have included sending tailored health messages to college students who are trying to quit smoking. This research brief…

DCS: A global satellite environmental data collection system study

NASA Technical Reports Server (NTRS)

1973-01-01

Cost analysis and technical feasibility data are presented on five medium orbiting and six geosynchronous satellite data collection systems with varying degrees of spacecraft and local user terminal complexity. Data are also provided on system approaches, user requirements, and user classes. Systems considered include orbiting ERTS and EOS type satellites and geosynchronous SmS and SEOS type data collectors.

Mixotrophic and heterotrophic production of lipids and carbohydrates by a locally isolated microalga using wastewater as a growth medium.

PubMed

Nzayisenga, Jean Claude; Eriksson, Karolina; Sellstedt, Anita

2018-06-01

The biomass production and changes in biochemical composition of a locally isolated microalga (Chlorella sp.) were investigated in autotrophic, mixotrophic and heterotrophic conditions, using glucose or glycerol as carbon sources and municipal wastewater as the growth medium. Both standard methods and Multivariate Curve Resolution-Alternating Least Squares (MCR-ALS) analysis of data acquired by Fourier-transform IR (FTIR) spectrometry showed that autotrophic and mixotrophic conditions promoted carbohydrate accumulation, while heterotrophic conditions with glycerol resulted in the highest lipid content and lowest carbohydrate content. Heterotrophic conditions with glycerol as a carbon source also resulted in high oleic acid (18:1) contents and low linolenic acid (18:3) contents, and thus increasing biodiesel quality. The results also show the utility of MCR-ALS for analyzing changes in microalgal biochemical composition. Copyright © 2018 Elsevier Ltd. All rights reserved.

Use of Nuclepore filters for ambient and workplace nanoparticle exposure assessment-Spherical particles

NASA Astrophysics Data System (ADS)

Chen, Sheng-Chieh; Wang, Jing; Fissan, Heinz; Pui, David Y. H.

2013-10-01

Nuclepore filter collection with subsequent electron microscopy analysis for nanoparticles was carried out to examine the feasibility of the method to assess the nanoparticle exposure. The number distribution of nanoparticles collected on the filter surface was counted visually and converted to the distribution in the air using existing filtration models for Nuclepore filters. To search for a proper model, this paper studied the overall penetrations of three different nanoparticles (PSL, Ag and NaCl), covering a wide range of particle sizes (20-800 nm) and densities (1.05-10.5 g cm-3), through Nuclepore filters with two different pore diameters (1 and 3 μm) and different face velocities (2-15 cm s-1). The data were compared with existing particle deposition models and modified models proposed by this study, which delivered different results because of different deposition processes considered. It was found that a parameter associated with flow condition and filter geometry (density of fluid medium, particle density, filtration face velocity, filter porosity and pore diameter) should be taken into account to verify the applicability of the models. The data of the overall penetration were in very good agreement with the properly applied models. A good agreement of filter surface collection between the validated model and the SEM analysis was obtained, indicating a correct nanoparticle number distribution in the air can be converted from the Nuclepore filter surface collection and this method can be applied for nanoparticle exposure assessment.

Production of Mushroom Mycelium as a Protein and Fat Source in Submerged Culture in Medium of Vinasse

PubMed Central

Falanghe, H.

1962-01-01

Of ten mushroom cultures investigated, only Agaricus campestris, Boletus indecisus, and Tricholoma nudum were capable of growing in submerged culture in medium of vinasse with added salts. Higher fermentative efficiencies were found under these conditions than in medium containing molasses or waste sulfite liquor. A. campestris showed a better capacity to produce protein but, since B. indecisus is capable of developing greater mycelium weight, its fermentative efficiencies are comparable. Both microorganisms could be grown in medium of vinasse with greatly varied amounts, producing higher mycelial weight in media with greater vinasse. The capacity of B. indecisus and A. campestris to utilize the noncarbohydrate fraction in total solids, instead of the total carbohydrates when they are in smaller amount, was observed in medium containing vinasse. B. indecisus and A. campestris were easily separated by filtration from the medium, although T. nudum was difficult to separate by this procedure. In experiments with A. campestris, the adaptative capacity of the organism to vinasse was demonstrated. PMID:13962715

Cultivation of Arthrospira (spirulina) platensis in desalinator wastewater and salinated synthetic medium: protein content and amino-acid profile

PubMed Central

Volkmann, Harriet; Imianovsky, Ulisses; Oliveira, Jorge L.B.; Sant’Anna, Ernani S.

2008-01-01

Arthrospira (Spirulina) platensis was cultivated in laboratory under controlled conditions (30°C, photoperiod of 12 hours light/dark provided by fluorescent lamps at a light intensity of 140 μmol photons.m-2.s-1 and constant bubbling air) in three different culture media: (1) Paoletti medium (control), (2) Paoletti supplemented with 1 g.L-1 NaCl (salinated water) and (3) Paoletti medium prepared with desalinator wastewater. The effects of these treatments on growth, protein content and amino acid profile were measured. Maximum cell concentrations observed in Paoletti medium, Paoletti supplemented with salinated water or with desalinator wastewater were 2.587, 3.545 and 4.954 g.L-1, respectively. Biomass in medium 3 presented the highest protein content (56.17%), while biomass in medium 2 presented 48.59% protein. All essential amino acids, except lysine and tryptophan, were found in concentrations higher than those requiried by FAO. PMID:24031187

Medium Calcium Concentration Determines Keratin Intermediate Filament Density and Distribution in Immortalized Cultured Thymic Epithelial Cells (TECs)

NASA Astrophysics Data System (ADS)

Sands, Sandra S.; Meek, William D.; Hayashi, Jun; Ketchum, Robert J.

2005-08-01

Isolation and culture of thymic epithelial cells (TECs) using conventional primary tissue culture techniques under conditions employing supplemented low calcium medium yielded an immortalized cell line derived from the LDA rat (Lewis [Rt1l] cross DA [Rt1a]) that could be manipulated in vitro. Thymi were harvested from 4 5-day-old neonates, enzymically digested using collagenase (1 mg/ml, 37°C, 1 h) and cultured in low calcium WAJC404A medium containing cholera toxin (20 ng/ml), dexamethasone (10 nM), epidermal growth factor (10 ng/ml), insulin (10 [mu]g/ml), transferrin (10 [mu]g/ml), 2% calf serum, 2.5% Dulbecco's Modified Eagle's Medium (DMEM), and 1% antibiotic/antimycotic. TECs cultured in low calcium displayed round to spindle-shaped morphology, distinct intercellular spaces (even at confluence), and dense reticular-like keratin patterns. In high calcium (0.188 mM), TECs formed cobblestone-like confluent monolayers that were resistant to trypsinization (0.05%) and displayed keratin intermediate filaments concentrated at desmosomal junctions between contiguous cells. Changes in cultured TEC morphology were quantified by an analysis of desmosome/membrane relationships in high and low calcium media. Desmosomes were significantly increased in the high calcium medium. These studies may have value when considering the growth conditions of cultured primary cell lines like TECs.