WorldWideScience

Sample records for storage droplet proteins

  1. Hormone signaling linked to silkmoth sex pheromone biosynthesis involves Ca2+/calmodulin-dependent protein kinase II-mediated phosphorylation of the insect PAT family protein Bombyx mori lipid storage droplet protein-1(BmLsd)

    Science.gov (United States)

    The structurally-related members of the PAT family of proteins, which are so name based on similarity amongst perilipin, adipophilin/adipocyte differentiation-related protein (ADRP), and tail-interacting protein of 47 kilodaltons (TIP47), are cytoplasmic lipid droplet (LD)-associated proteins charac...

  2. Protein Correlation Profiles Identify Lipid Droplet Proteins with High Confidence*

    Science.gov (United States)

    Krahmer, Natalie; Hilger, Maximiliane; Kory, Nora; Wilfling, Florian; Stoehr, Gabriele; Mann, Matthias; Farese, Robert V.; Walther, Tobias C.

    2013-01-01

    Lipid droplets (LDs) are important organelles in energy metabolism and lipid storage. Their cores are composed of neutral lipids that form a hydrophobic phase and are surrounded by a phospholipid monolayer that harbors specific proteins. Most well-established LD proteins perform important functions, particularly in cellular lipid metabolism. Morphological studies show LDs in close proximity to and interacting with membrane-bound cellular organelles, including the endoplasmic reticulum, mitochondria, peroxisomes, and endosomes. Because of these close associations, it is difficult to purify LDs to homogeneity. Consequently, the confident identification of bona fide LD proteins via proteomics has been challenging. Here, we report a methodology for LD protein identification based on mass spectrometry and protein correlation profiles. Using LD purification and quantitative, high-resolution mass spectrometry, we identified LD proteins by correlating their purification profiles to those of known LD proteins. Application of the protein correlation profile strategy to LDs isolated from Drosophila S2 cells led to the identification of 111 LD proteins in a cellular LD fraction in which 1481 proteins were detected. LD localization was confirmed in a subset of identified proteins via microscopy of the expressed proteins, thereby validating the approach. Among the identified LD proteins were both well-characterized LD proteins and proteins not previously known to be localized to LDs. Our method provides a high-confidence LD proteome of Drosophila cells and a novel approach that can be applied to identify LD proteins of other cell types and tissues. PMID:23319140

  3. Lipid droplets as ubiquitous fat storage organelles in C. elegans

    Directory of Open Access Journals (Sweden)

    Guo Fengli

    2010-12-01

    Full Text Available Abstract Background Lipid droplets are a class of eukaryotic cell organelles for storage of neutral fat such as triacylglycerol (TAG and cholesterol ester (CE. We and others have recently reported that lysosome-related organelles (LROs are not fat storage structures in the nematode C. elegans. We also reported the formation of enlarged lipid droplets in a class of peroxisomal fatty acid β-oxidation mutants. In the present study, we seek to provide further evidence on the organelle nature and biophysical properties of fat storage structures in wild-type and mutant C. elegans. Results In this study, we provide biochemical, histological and ultrastructural evidence of lipid droplets in wild-type and mutant C. elegans that lack lysosome related organelles (LROs. The formation of lipid droplets and the targeting of BODIPY fatty acid analogs to lipid droplets in live animals are not dependent on lysosomal trafficking or peroxisome dysfunction. However, the targeting of Nile Red to lipid droplets in live animals occurs only in mutants with defective peroxisomes. Nile Red labelled-lipid droplets are characterized by a fluorescence emission spectrum distinct from that of Nile Red labelled-LROs. Moreover, we show that the recently developed post-fix Nile Red staining method labels lipid droplets exclusively. Conclusions Our results demonstrate lipid droplets as ubiquitous fat storage organelles and provide a unified explanation for previous studies on fat labelling methods in C. elegans. These results have important applications to the studies of fat storage and lipid droplet regulation in the powerful genetic system, C. elegans.

  4. CaMKII-MEDIATED PHOSPHORYLATION OF THE BOMBYX MORI LIPID STORAGE DROPLET PROTEIN-1 (BmLsd1), AN INSECT PAT FAMILY PROTEIN, IS ESSENTIAL FOR SILKMOTH SEX PHEROMONE BIOSYNTHESIS

    Science.gov (United States)

    The structurally-related members of the PAT family of proteins, which are so name based on similarity amongst perilipin, adipophilin/adipocyte differentiation-related protein (ADRP), and tail-interacting protein of 47 kilodaltons (TIP47), are cytoplasmic lipid droplet (LD)-associated proteins charac...

  5. Rheological properties of soybean protein isolate gels containing emulsion droplets

    NARCIS (Netherlands)

    Kim, K.H.; Renkema, J.M.S.; Vliet, van T.

    2001-01-01

    Rheological properties of soybean protein gels containing various volume fractions oil droplets have been studied at small and large deformations. Dynamic viscoelastic properties of soybean protein isolate gels were determined as a function of the volume fraction of oil droplets stabilised by the

  6. Droplet networks with incorporated protein diodes show collective properties

    Science.gov (United States)

    Maglia, Giovanni; Heron, Andrew J.; Hwang, William L.; Holden, Matthew A.; Mikhailova, Ellina; Li, Qiuhong; Cheley, Stephen; Bayley, Hagan

    2009-07-01

    Recently, we demonstrated that submicrolitre aqueous droplets submerged in an apolar liquid containing lipid can be tightly connected by means of lipid bilayers to form networks. Droplet interface bilayers have been used for rapid screening of membrane proteins and to form asymmetric bilayers with which to examine the fundamental properties of channels and pores. Networks, meanwhile, have been used to form microscale batteries and to detect light. Here, we develop an engineered protein pore with diode-like properties that can be incorporated into droplet interface bilayers in droplet networks to form devices with electrical properties including those of a current limiter, a half-wave rectifier and a full-wave rectifier. The droplet approach, which uses unsophisticated components (oil, lipid, salt water and a simple pore), can therefore be used to create multidroplet networks with collective properties that cannot be produced by droplet pairs.

  7. Triacylglycerol Storage in Lipid Droplets in Procyclic Trypanosoma brucei.

    Directory of Open Access Journals (Sweden)

    Stefan Allmann

    Full Text Available Carbon storage is likely to enable adaptation of trypanosomes to nutritional challenges or bottlenecks during their stage development and migration in the tsetse. Lipid droplets are candidates for this function. This report shows that feeding of T. brucei with oleate results in a 4-5 fold increase in the number of lipid droplets, as quantified by confocal fluorescence microscopy and by flow cytometry of BODIPY 493/503-stained cells. The triacylglycerol (TAG content also increased 4-5 fold, and labeled oleate is incorporated into TAG. Fatty acid carbon can thus be stored as TAG in lipid droplets under physiological growth conditions in procyclic T. brucei. β-oxidation has been suggested as a possible catabolic pathway for lipids in T. brucei. A single candidate gene, TFEα1 with coding capacity for a subunit of the trifunctional enzyme complex was identified. TFEα1 is expressed in procyclic T. brucei and present in glycosomal proteomes, Unexpectedly, a TFEα1 gene knock-out mutant still expressed wild-type levels of previously reported NADP-dependent 3-hydroxyacyl-CoA dehydrogenase activity, and therefore, another gene encodes this enzymatic activity. Homozygous Δtfeα1/Δtfeα1 null mutant cells show a normal growth rate and an unchanged glycosomal proteome in procyclic T. brucei. The decay kinetics of accumulated lipid droplets upon oleate withdrawal can be fully accounted for by the dilution effect of cell division in wild-type and Δtfeα1/Δtfeα1 cells. The absence of net catabolism of stored TAG in procyclic T. brucei, even under strictly glucose-free conditions, does not formally exclude a flux through TAG, in which biosynthesis equals catabolism. Also, the possibility remains that TAG catabolism is completely repressed by other carbon sources in culture media or developmentally activated in post-procyclic stages in the tsetse.

  8. Tuber storage proteins.

    Science.gov (United States)

    Shewry, Peter R

    2003-06-01

    A wide range of plants are grown for their edible tubers, but five species together account for almost 90 % of the total world production. These are potato (Solanum tuberosum), cassava (Manihot esculenta), sweet potato (Ipomoea batatus), yams (Dioscorea spp.) and taro (Colocasia, Cyrtosperma and Xanthosoma spp.). All of these, except cassava, contain groups of storage proteins, but these differ in the biological properties and evolutionary relationships. Thus, patatin from potato exhibits activity as an acylhydrolase and esterase, sporamin from sweet potato is an inhibitor of trypsin, and dioscorin from yam is a carbonic anhydrase. Both sporamin and dioscorin also exhibit antioxidant and radical scavenging activity. Taro differs from the other three crops in that it contains two major types of storage protein: a trypsin inhibitor related to sporamin and a mannose-binding lectin. These characteristics indicate that tuber storage proteins have evolved independently in different species, which contrasts with the highly conserved families of storage proteins present in seeds. Furthermore, all exhibit biological activities which could contribute to resistance to pests, pathogens or abiotic stresses, indicating that they may have dual roles in the tubers.

  9. Quantification of protein interaction kinetics in a micro droplet

    Energy Technology Data Exchange (ETDEWEB)

    Yin, L. L. [Center for Bioelectronics and Biosensors, Biodesign Institute, Arizona State University, Tempe, Arizona 85287 (United States); College of Chemistry and Chemical Engineering, Chongqing University, Chongqing 400044 (China); Wang, S. P., E-mail: shaopeng.wang@asu.edu, E-mail: njtao@asu.edu; Shan, X. N.; Tao, N. J., E-mail: shaopeng.wang@asu.edu, E-mail: njtao@asu.edu [Center for Bioelectronics and Biosensors, Biodesign Institute, Arizona State University, Tempe, Arizona 85287 (United States); Zhang, S. T. [College of Chemistry and Chemical Engineering, Chongqing University, Chongqing 400044 (China)

    2015-11-15

    Characterization of protein interactions is essential to the discovery of disease biomarkers, the development of diagnostic assays, and the screening for therapeutic drugs. Conventional flow-through kinetic measurements need relative large amount of sample that is not feasible for precious protein samples. We report a novel method to measure protein interaction kinetics in a single droplet with sub microliter or less volume. A droplet in a humidity-controlled environmental chamber is replacing the microfluidic channels as the reactor for the protein interaction. The binding process is monitored by a surface plasmon resonance imaging (SPRi) system. Association curves are obtained from the average SPR image intensity in the center area of the droplet. The washing step required by conventional flow-through SPR method is eliminated in the droplet method. The association and dissociation rate constants and binding affinity of an antigen-antibody interaction are obtained by global fitting of association curves at different concentrations. The result obtained by this method is accurate as validated by conventional flow-through SPR system. This droplet-based method not only allows kinetic studies for proteins with limited supply but also opens the door for high-throughput protein interaction study in a droplet-based microarray format that enables measurement of many to many interactions on a single chip.

  10. Tuber Storage Proteins

    OpenAIRE

    SHEWRY, PETER R.

    2003-01-01

    A wide range of plants are grown for their edible tubers, but five species together account for almost 90 % of the total world production. These are potato (Solanum tuberosum), cassava (Manihot esculenta), sweet potato (Ipomoea batatus), yams (Dioscorea spp.) and taro (Colocasia, Cyrtosperma and Xanthosoma spp.). All of these, except cassava, contain groups of storage proteins, but these differ in the biological properties and evolutionary relationships. Thus, patatin from potato exhibits act...

  11. Addressable droplet microarrays for single cell protein analysis.

    Science.gov (United States)

    Salehi-Reyhani, Ali; Burgin, Edward; Ces, Oscar; Willison, Keith R; Klug, David R

    2014-11-07

    Addressable droplet microarrays are potentially attractive as a way to achieve miniaturised, reduced volume, high sensitivity analyses without the need to fabricate microfluidic devices or small volume chambers. We report a practical method for producing oil-encapsulated addressable droplet microarrays which can be used for such analyses. To demonstrate their utility, we undertake a series of single cell analyses, to determine the variation in copy number of p53 proteins in cells of a human cancer cell line.

  12. Lipid droplet meets a mitochondrial protein to regulate adipocyte lipolysis

    Science.gov (United States)

    In response to adrenergic stimulation, adipocytes undergo protein kinase A (PKA)-stimulated lipolysis. A key PKA target in this context is perilipin 1, a major regulator of lipolysis on lipid droplets (LDs). A study published in this issue of The EMBO Journal (Pidoux et al, 2011) identifies optic at...

  13. Omic studies reveal the pathogenic lipid droplet proteins in non-alcoholic fatty liver disease

    Directory of Open Access Journals (Sweden)

    Xuelin Zhang

    2016-10-01

    Full Text Available Abstract Non-alcoholic fatty liver disease (NAFLD is an epidemic metabolic condition driven by an underlying lipid homeostasis disorder. The lipid droplet (LD, the main organelle involved in neutral lipid storage and hydrolysis, is a potential target for NAFLD therapeutic treatment. In this review, we summarize recent progress elucidating the connections between LD-associated proteins and NAFLD found by genome-wide association studies (GWAS, genomic and proteomic studies. Finally, we discuss a possible mechanism by which the protein 17β-hydroxysteroid dehydrogenase 13 (17β-HSD13 may promote the development of NAFLD.

  14. Structural Insights into Triglyceride Storage Mediated by Fat Storage-Inducing Transmembrane (FIT) Protein 2

    Science.gov (United States)

    Gross, David A.; Snapp, Erik L.; Silver, David L.

    2010-01-01

    Fat storage-Inducing Transmembrane proteins 1 & 2 (FIT1/FITM1 and FIT2/FITM2) belong to a unique family of evolutionarily conserved proteins localized to the endoplasmic reticulum that are involved in triglyceride lipid droplet formation. FIT proteins have been shown to mediate the partitioning of cellular triglyceride into lipid droplets, but not triglyceride biosynthesis. FIT proteins do not share primary sequence homology with known proteins and no structural information is available to inform on the mechanism by which FIT proteins function. Here, we present the experimentally-solved topological models for FIT1 and FIT2 using N-glycosylation site mapping and indirect immunofluorescence techniques. These methods indicate that both proteins have six-transmembrane-domains with both N- and C-termini localized to the cytosol. Utilizing this model for structure-function analysis, we identified and characterized a gain-of-function mutant of FIT2 (FLL(157-9)AAA) in transmembrane domain 4 that markedly augmented the total number and mean size of lipid droplets. Using limited-trypsin proteolysis we determined that the FLL(157-9)AAA mutant has enhanced trypsin cleavage at K86 relative to wild-type FIT2, indicating a conformational change. Taken together, these studies indicate that FIT2 is a 6 transmembrane domain-containing protein whose conformation likely regulates its activity in mediating lipid droplet formation. PMID:20520733

  15. Structural insights into triglyceride storage mediated by fat storage-inducing transmembrane (FIT protein 2.

    Directory of Open Access Journals (Sweden)

    David A Gross

    2010-05-01

    Full Text Available Fat storage-Inducing Transmembrane proteins 1 & 2 (FIT1/FITM1 and FIT2/FITM2 belong to a unique family of evolutionarily conserved proteins localized to the endoplasmic reticulum that are involved in triglyceride lipid droplet formation. FIT proteins have been shown to mediate the partitioning of cellular triglyceride into lipid droplets, but not triglyceride biosynthesis. FIT proteins do not share primary sequence homology with known proteins and no structural information is available to inform on the mechanism by which FIT proteins function. Here, we present the experimentally-solved topological models for FIT1 and FIT2 using N-glycosylation site mapping and indirect immunofluorescence techniques. These methods indicate that both proteins have six-transmembrane-domains with both N- and C-termini localized to the cytosol. Utilizing this model for structure-function analysis, we identified and characterized a gain-of-function mutant of FIT2 (FLL(157-9AAA in transmembrane domain 4 that markedly augmented the total number and mean size of lipid droplets. Using limited-trypsin proteolysis we determined that the FLL(157-9AAA mutant has enhanced trypsin cleavage at K86 relative to wild-type FIT2, indicating a conformational change. Taken together, these studies indicate that FIT2 is a 6 transmembrane domain-containing protein whose conformation likely regulates its activity in mediating lipid droplet formation.

  16. Identification of a new class of lipid droplet-associated proteins in plants

    Science.gov (United States)

    Lipid droplets in plants (also known as oil bodies, lipid bodies or oleosomes) are well characterized in seeds, and oleosins, the major proteins associated with their surface, were shown to be important for stabilizing lipid droplets during seed desiccation and rehydration. However, lipid droplets ...

  17. Hepatitis C Virus Core Protein Decreases Lipid Droplet Turnover

    Science.gov (United States)

    Harris, Charles; Herker, Eva; Farese, Robert V.; Ott, Melanie

    2011-01-01

    Steatosis is a frequent complication of hepatitis C virus infection. In mice, this condition is recapitulated by the expression of a single viral protein, the nucleocapsid core. Core localizes to the surface of lipid droplets (LDs) in infected liver cells through a process dependent on host diacylglycerol acyltransferase 1 (DGAT1), an enzyme that synthesizes triglycerides in the endoplasmic reticulum. Whether DGAT1 also plays a role in core-induced steatosis is uncertain. Here, we show that mouse embryonic fibroblasts isolated from DGAT1−/− mice are protected from core-induced steatosis, as are livers of DGAT1−/− mice expressing core, demonstrating that the steatosis is DGAT1-dependent. Surprisingly, core expression did not increase DGAT1 activity or triglyceride synthesis, thus excluding the possibility that core activates DGAT1 to cause steatosis. Instead, we find that DGAT1-dependent localization of core to LDs is a prerequisite for the steatogenic properties of the core. Using biochemical and immunofluorescence microscopy techniques, we show that the turnover of lipids in core-coated droplets is decreased, providing a physiological mechanism for core-induced steatosis. Our results support a bipartite model in which core first requires DGAT1 to gain access to LDs, and then LD-localized core interferes with triglyceride turnover, thus stabilizing lipid droplets and leading to steatosis. PMID:21984835

  18. Apoptosis inhibitor of macrophage (AIM) diminishes lipid droplet-coating proteins leading to lipolysis in adipocytes

    International Nuclear Information System (INIS)

    Iwamura, Yoshihiro; Mori, Mayumi; Nakashima, Katsuhiko; Mikami, Toshiyuki; Murayama, Katsuhisa; Arai, Satoko; Miyazaki, Toru

    2012-01-01

    Highlights: ► AIM induces lipolysis in a distinct manner from that of hormone-dependent lipolysis. ► AIM ablates activity of peroxisome proliferator-activated receptor in adipocytes. ► AIM reduces mRNA levels of lipid-droplet coating proteins leading to lipolysis. -- Abstract: Under fasting conditions, triacylglycerol in adipose tissue undergoes lipolysis to supply fatty acids as energy substrates. Such lipolysis is regulated by hormones, which activate lipases via stimulation of specific signalling cascades. We previously showed that macrophage-derived soluble protein, AIM induces obesity-associated lipolysis, triggering chronic inflammation in fat tissue which causes insulin resistance. However, the mechanism of how AIM mediates lipolysis remains unknown. Here we show that AIM induces lipolysis in a manner distinct from that of hormone-dependent lipolysis, without activation or augmentation of lipases. In vivo and in vitro, AIM did not enhance phosphorylation of hormone-sensitive lipase (HSL) in adipocytes, a hallmark of hormone-dependent lipolysis activation. Similarly, adipose tissue from obese AIM-deficient and wild-type mice showed comparable HSL phosphorylation. Consistent with the suppressive effect of AIM on fatty acid synthase activity, the amount of saturated and unsaturated fatty acids was reduced in adipocytes treated with AIM. This response ablated transcriptional activity of peroxisome proliferator-activated receptor (PPARγ), leading to diminished gene expression of lipid-droplet coating proteins including fat-specific protein 27 (FSP27) and Perilipin, which are indispensable for triacylglycerol storage in adipocytes. Accordingly, the lipolytic effect of AIM was overcome by a PPARγ-agonist or forced expression of FSP27, while it was synergized by a PPARγ-antagonist. Overall, distinct modes of lipolysis appear to take place in different physiological situations; one is a supportive response against nutritional deprivation achieved by

  19. Lipidomic and proteomic analysis of Caenorhabditis elegans lipid droplets and identification of ACS-4 as a lipid droplet-associated protein

    Energy Technology Data Exchange (ETDEWEB)

    Vrablik, Tracy L. [Washington State Univ., Pullman, WA (United States); Petyuk, Vladislav A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Larson, Emily M. [Washington State Univ., Pullman, WA (United States); Smith, Richard D. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Watts, Jennifer [Washington State Univ., Pullman, WA (United States)

    2015-06-27

    Lipid droplets are cytoplasmic organelles that store neutral lipids for membrane synthesis and energy reserves. In this study, we characterized the lipid and protein composition of purified C. elegans lipid droplets. These lipid droplets are composed mainly of triacylglycerols, surrounded by a phospholipid monolayer composed primarily of phosphatidylcholine and phosphatidylethanolamine. The fatty acid composition of the triacylglycerols was rich in fatty acid species obtained from the dietary E. coli, including cyclopropane fatty acids and cis-vaccenic acid. Unlike other organisms, C. elegans lipid droplets contain very little cholesterol or cholesterol esters. Comparison of the lipid droplet proteomes of wild type and high-fat daf-2 mutant strains shows a relative decrease of MDT-28 abundance in lipid droplets isolated from daf-2 mutants. Functional analysis of lipid droplet proteins identified in our proteomic studies indicated an enrichment of proteins required for growth and fat homeostasis in C. elegans.

  20. The adaptor protein alpha-syntrophin regulates adipocyte lipid droplet growth

    Energy Technology Data Exchange (ETDEWEB)

    Eisinger, Kristina; Rein-Fischboeck, Lisa; Pohl, Rebekka; Meier, Elisabeth M.; Krautbauer, Sabrina; Buechler, Christa, E-mail: christa.buechler@klinik.uni-regensburg.de

    2016-07-01

    The scaffold protein alpha-syntrophin (SNTA) regulates lipolysis indicating a role in lipid homeostasis. Adipocytes are the main lipid storage cells in the body, and here, the function of SNTA has been analyzed in 3T3-L1 cells. SNTA is expressed in preadipocytes and is induced early during adipogenesis. Knock-down of SNTA in preadipocytes increases their proliferation. Proteins which are induced during adipogenesis like adiponectin and caveolin-1, and the inflammatory cytokine IL-6 are at normal levels in the mature cells differentiated from preadipocytes with low SNTA. This suggests that SNTA does neither affect differentiation nor inflammation. Expression of proteins with a role in cholesterol and triglyceride homeostasis is unchanged. Consequently, basal and epinephrine induced lipolysis as well as insulin stimulated phosphorylation of Akt and ERK1/2 are normal. Importantly, adipocytes with low SNTA form smaller lipid droplets and store less triglycerides. Stearoyl-CoA reductase and MnSOD are reduced upon SNTA knock-down but do not contribute to lower lipid levels. Oleate uptake is even increased in cells with SNTA knock-down. In summary, current data show that SNTA is involved in the expansion of lipid droplets independent of adipogenesis. Enhanced preadipocyte proliferation and capacity to store surplus fatty acids may protect adipocytes with low SNTA from lipotoxicity in obesity. - Highlights: • Alpha-syntrophin (SNTA) is expressed in 3T3-L1adipocytes. • SNTA knock-down in preadipocytes has no effect on adipogenesis. • Mature 3T3-L1 differentiated from cells with low SNTA form small lipid droplets. • SCD1 and MnSOD are reduced in adipocytes with low SNTA. • SCD1 knock-down does not alter triglyceride levels.

  1. Arabidopsis SEIPIN Proteins Modulate Triacylglycerol Accumulation and Influence Lipid Droplet Proliferation[OPEN

    Science.gov (United States)

    2015-01-01

    The lipodystrophy protein SEIPIN is important for lipid droplet (LD) biogenesis in human and yeast cells. In contrast with the single SEIPIN genes in humans and yeast, there are three SEIPIN homologs in Arabidopsis thaliana, designated SEIPIN1, SEIPIN2, and SEIPIN3. Essentially nothing is known about the functions of SEIPIN homologs in plants. Here, a yeast (Saccharomyces cerevisiae) SEIPIN deletion mutant strain and a plant (Nicotiana benthamiana) transient expression system were used to test the ability of Arabidopsis SEIPINs to influence LD morphology. In both species, expression of SEIPIN1 promoted accumulation of large-sized lipid droplets, while expression of SEIPIN2 and especially SEIPIN3 promoted small LDs. Arabidopsis SEIPINs increased triacylglycerol levels and altered composition. In tobacco, endoplasmic reticulum (ER)-localized SEIPINs reorganized the normal, reticulated ER structure into discrete ER domains that colocalized with LDs. N-terminal deletions and swapping experiments of SEIPIN1 and 3 revealed that this region of SEIPIN determines LD size. Ectopic overexpression of SEIPIN1 in Arabidopsis resulted in increased numbers of large LDs in leaves, as well as in seeds, and increased seed oil content by up to 10% over wild-type seeds. By contrast, RNAi suppression of SEIPIN1 resulted in smaller seeds and, as a consequence, a reduction in the amount of oil per seed compared with the wild type. Overall, our results indicate that Arabidopsis SEIPINs are part of a conserved LD biogenesis machinery in eukaryotes and that in plants these proteins may have evolved specialized roles in the storage of neutral lipids by differentially modulating the number and sizes of lipid droplets. PMID:26362606

  2. Lipid droplet-associated proteins in alcoholic liver disease: a potential linkage with hepatocellular damage

    OpenAIRE

    Ikura, Yoshihiro; Caldwell, Stephen H

    2015-01-01

    Steatosis is a characteristic morphological change of alcoholic liver disease, but its pathologic significance is still obscure. Regardless of cell types, intracellular lipid droplets are coated with a phospholipid monolayer, on which many kinds of lipid droplet-associated proteins are present. These proteins, such as the perilipin family of proteins and the cell death inducing DNA fragmentation factor (DFF) 45-like effectors, are recognized to play important roles in lipid metabolism in the ...

  3. Apoptosis inhibitor of macrophage (AIM) diminishes lipid droplet-coating proteins leading to lipolysis in adipocytes

    Energy Technology Data Exchange (ETDEWEB)

    Iwamura, Yoshihiro; Mori, Mayumi; Nakashima, Katsuhiko [Laboratory of Molecular Biomedicine for Pathogenesis, Center for Disease Biology and Integrative Medicine, Faculty of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033 (Japan); Mikami, Toshiyuki; Murayama, Katsuhisa [Genomic Science Laboratories, Dainippon Sumitomo Pharma Co. Ltd., 3-1-98 Kasugadenaka, Konohana-ku, Osaka 554-0022 (Japan); Arai, Satoko [Laboratory of Molecular Biomedicine for Pathogenesis, Center for Disease Biology and Integrative Medicine, Faculty of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033 (Japan); Miyazaki, Toru, E-mail: tm@m.u-tokyo.ac.jp [Laboratory of Molecular Biomedicine for Pathogenesis, Center for Disease Biology and Integrative Medicine, Faculty of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033 (Japan)

    2012-06-08

    Highlights: Black-Right-Pointing-Pointer AIM induces lipolysis in a distinct manner from that of hormone-dependent lipolysis. Black-Right-Pointing-Pointer AIM ablates activity of peroxisome proliferator-activated receptor in adipocytes. Black-Right-Pointing-Pointer AIM reduces mRNA levels of lipid-droplet coating proteins leading to lipolysis. -- Abstract: Under fasting conditions, triacylglycerol in adipose tissue undergoes lipolysis to supply fatty acids as energy substrates. Such lipolysis is regulated by hormones, which activate lipases via stimulation of specific signalling cascades. We previously showed that macrophage-derived soluble protein, AIM induces obesity-associated lipolysis, triggering chronic inflammation in fat tissue which causes insulin resistance. However, the mechanism of how AIM mediates lipolysis remains unknown. Here we show that AIM induces lipolysis in a manner distinct from that of hormone-dependent lipolysis, without activation or augmentation of lipases. In vivo and in vitro, AIM did not enhance phosphorylation of hormone-sensitive lipase (HSL) in adipocytes, a hallmark of hormone-dependent lipolysis activation. Similarly, adipose tissue from obese AIM-deficient and wild-type mice showed comparable HSL phosphorylation. Consistent with the suppressive effect of AIM on fatty acid synthase activity, the amount of saturated and unsaturated fatty acids was reduced in adipocytes treated with AIM. This response ablated transcriptional activity of peroxisome proliferator-activated receptor (PPAR{gamma}), leading to diminished gene expression of lipid-droplet coating proteins including fat-specific protein 27 (FSP27) and Perilipin, which are indispensable for triacylglycerol storage in adipocytes. Accordingly, the lipolytic effect of AIM was overcome by a PPAR{gamma}-agonist or forced expression of FSP27, while it was synergized by a PPAR{gamma}-antagonist. Overall, distinct modes of lipolysis appear to take place in different physiological

  4. Fat storage-inducing transmembrane protein 2 is required for normal fat storage in adipose tissue.

    Science.gov (United States)

    Miranda, Diego A; Kim, Ji-Hyun; Nguyen, Long N; Cheng, Wang; Tan, Bryan C; Goh, Vera J; Tan, Jolene S Y; Yaligar, Jadegoud; Kn, Bhanu Prakash; Velan, S Sendhil; Wang, Hongyan; Silver, David L

    2014-04-04

    Triglycerides within the cytosol of cells are stored in a phylogenetically conserved organelle called the lipid droplet (LD). LDs can be formed at the endoplasmic reticulum, but mechanisms that regulate the formation of LDs are incompletely understood. Adipose tissue has a high capacity to form lipid droplets and store triglycerides. Fat storage-inducing transmembrane protein 2 (FITM2/FIT2) is highly expressed in adipocytes, and data indicate that FIT2 has an important role in the formation of LDs in cells, but whether FIT2 has a physiological role in triglyceride storage in adipose tissue remains unproven. Here we show that adipose-specific deficiency of FIT2 (AF2KO) in mice results in progressive lipodystrophy of white adipose depots and metabolic dysfunction. In contrast, interscapular brown adipose tissue of AF2KO mice accumulated few but large LDs without changes in cellular triglyceride levels. High fat feeding of AF2KO mice or AF2KO mice on the genetically obese ob/ob background accelerated the onset of lipodystrophy. At the cellular level, primary adipocyte precursors of white and brown adipose tissue differentiated in vitro produced fewer but larger LDs without changes in total cellular triglyceride or triglyceride biosynthesis. These data support the conclusion that FIT2 plays an essential, physiological role in fat storage in vivo.

  5. Liver X Receptors Balance Lipid Stores in Hepatic Stellate Cells via Rab18, a Retinoid Responsive Lipid Droplet Protein

    Science.gov (United States)

    O’Mahony, Fiona; Wroblewski, Kevin; O’Byrne, Sheila M.; Jiang, Hongfeng; Clerkin, Kara; Benhammou, Jihane; Blaner, William S.; Beaven, Simon W.

    2014-01-01

    Liver X receptors (LXRs) are determinants of hepatic stellate cell (HSC) activation and liver fibrosis. Freshly isolated HSCs from Lxrαβ−/− mice have increased lipid droplet (LD) size but the functional consequences of this are unknown. Our aim was to determine whether LXRs link cholesterol to retinoid storage in HSCs and how this impacts activation. Primary HSCs from Lxrαβ−/− and wild-type (WT) mice were profiled by gene array during in vitro activation. Lipid content was quantified by HPLC and mass spectroscopy. Primary HSCs were treated with nuclear receptor ligands, transfected with siRNA and plasmid constructs, and analyzed by immunocytochemistry. Lxrαβ−/− HSCs have increased cholesterol and retinyl esters (CEs & REs). The retinoid increase drives intrinsic retinoic acid receptor (RAR) signaling and activation occurs more rapidly in Lxrαβ−/− HSCs. We identify Rab18 as a novel retinoic acid responsive, lipid droplet associated protein that helps mediate stellate cell activation. Rab18 mRNA, protein, and membrane insertion increase during activation. Both Rab18 GTPase activity and isoprenylation are required for stellate cell lipid droplet loss and induction of activation markers. These phenomena are accelerated in the Lxrαβ−/− HSCs, where there is greater retinoic acid flux. Conversely, Rab18 knockdown retards lipid droplet loss in culture and blocks activation, just like the functional mutants. Rab18 is also induced with acute liver injury in vivo. Conclusion Retinoid and cholesterol metabolism are linked in stellate cells by the LD associated protein, Rab18. Retinoid overload helps explain the pro-fibrotic phenotype of Lxrαβ−/− mice and we establish a pivotal role for Rab18 GTPase activity and membrane insertion in wild-type stellate cell activation. Interference with Rab18 may have significant therapeutic benefit in ameliorating liver fibrosis. PMID:25482505

  6. Electrospray droplet exposure to organic vapors: metal ion removal from proteins and protein complexes.

    Science.gov (United States)

    DeMuth, J Corinne; McLuckey, Scott A

    2015-01-20

    The exposure of aqueous nanoelectrospray droplets to various organic vapors can dramatically reduce sodium adduction on protein ions in positive ion mass spectra. Volatile alcohols, such as methanol, ethanol, and isopropanol lead to a significant reduction in sodium ion adduction but are not as effective as acetonitrile, acetone, and ethyl acetate. Organic vapor exposure in the negative ion mode, on the other hand, has essentially no effect on alkali ion adduction. Evidence is presented to suggest that the mechanism by which organic vapor exposure reduces alkali ion adduction in the positive mode involves the depletion of alkali metal ions via ion evaporation of metal ions solvated with organic molecules. The early generation of metal/organic cluster ions during the droplet desolvation process results in fewer metal ions available to condense on the protein ions formed via the charged residue mechanism. These effects are demonstrated with holomyoglobin ions to illustrate that the metal ion reduction takes place without detectable protein denaturation, which might be revealed by heme loss or an increase in charge state distribution. No evidence is observed for denaturation with exposure to any of the organic vapors evaluated in this work.

  7. High confidence proteomic analysis of yeast LDs identifies additional droplet proteins and reveals connections to dolichol synthesis and sterol acetylation.

    Science.gov (United States)

    Currie, Erin; Guo, Xiuling; Christiano, Romain; Chitraju, Chandramohan; Kory, Nora; Harrison, Kenneth; Haas, Joel; Walther, Tobias C; Farese, Robert V

    2014-07-01

    Accurate protein inventories are essential for understanding an organelle's functions. The lipid droplet (LD) is a ubiquitous intracellular organelle with major functions in lipid storage and metabolism. LDs differ from other organelles because they are bounded by a surface monolayer, presenting unique features for protein targeting to LDs. Many proteins of varied functions have been found in purified LD fractions by proteomics. While these studies have become increasingly sensitive, it is often unclear which of the identified proteins are specific to LDs. Here we used protein correlation profiling to identify 35 proteins that specifically enrich with LD fractions of Saccharomyces cerevisiae Of these candidates, 30 fluorophore-tagged proteins localize to LDs by microscopy, including six proteins, several with human orthologs linked to diseases, which we newly identify as LD proteins (Cab5, Rer2, Say1, Tsc10, YKL047W, and YPR147C). Two of these proteins, Say1, a sterol deacetylase, and Rer2, a cis-isoprenyl transferase, are enzymes involved in sterol and polyprenol metabolism, respectively, and we show their activities are present in LD fractions. Our results provide a highly specific list of yeast LD proteins and reveal that the vast majority of these proteins are involved in lipid metabolism. Copyright © 2014 by the American Society for Biochemistry and Molecular Biology, Inc.

  8. Enhancing physicochemical properties of emulsions by heteroaggregation of oppositely charged lactoferrin coated lutein droplets and whey protein isolate coated DHA droplets.

    Science.gov (United States)

    Li, Xin; Wang, Xu; Xu, Duoxia; Cao, Yanping; Wang, Shaojia; Wang, Bei; Sun, Baoguo; Yuan, Fang; Gao, Yanxiang

    2018-01-15

    The formation and physicochemical stability of mixed functional components (lutein & DHA) emulsions through heteroaggregation were studied. It was formed by controlled heteroaggregation of oppositely charged lutein and DHA droplets coated by cationic lactoferrin (LF) and anionic whey protein isolate (WPI), respectively. Heteroaggregation was induced by mixing the oppositely charged LF-lutein and WPI-DHA emulsions together at pH 6.0. Droplet size, zeta-potential, transmission-physical stability, microrheological behavior and microstructure of the heteroaggregates formed were measured as a function of LF-lutein to WPI-DHA droplet ratio. Lutein degradation and DHA oxidation by measurement of lipid hydroperoxides and thiobarbituric acid reactive substances were determined. Upon mixing the two types of bioactive compounds droplets together, it was found that the largest aggregates and highest physical stability occurred at a droplet ratio of 40% LF-lutein droplets to 60% WPI-DHA droplets. Heteroaggregates formation altered the microrheological properties of the mixed emulsions mainly by the special network structure of the droplets. When LF-coated lutein droplets ratios were more than 30% and less than 60%, the mixed emulsions exhibited distinct decreases in the Mean Square Displacement, which indicated that their limited scope of Brownian motion and stable structure. Mixed emulsions with LF-lutein/WPI-DHA droplets ratio of 4:6 exhibited Macroscopic Viscosity Index with 13 times and Elasticity Index with 3 times of magnitudes higher than the individual emulsions from which they were prepared. Compared with the WPI-DHA emulsion or LF-lutein emulsion, the oxidative stability of the heteroaggregate of LF-lutein/WPI-DHA emulsions was improved. Heteroaggregates formed by oppositely charged bioactive compounds droplets may be useful for creating specific food structures that lead to desirable physicochemical properties, such as microrheological property, physical and chemical

  9. A new fluorescence-based method identifies protein phosphatases regulating lipid droplet metabolism.

    Directory of Open Access Journals (Sweden)

    Bruno L Bozaquel-Morais

    Full Text Available In virtually every cell, neutral lipids are stored in cytoplasmic structures called lipid droplets (LDs and also referred to as lipid bodies or lipid particles. We developed a rapid high-throughput assay based on the recovery of quenched BODIPY-fluorescence that allows to quantify lipid droplets. The method was validated by monitoring lipid droplet turnover during growth of a yeast culture and by screening a group of strains deleted in genes known to be involved in lipid metabolism. In both tests, the fluorimetric assay showed high sensitivity and good agreement with previously reported data using microscopy. We used this method for high-throughput identification of protein phosphatases involved in lipid droplet metabolism. From 65 yeast knockout strains encoding protein phosphatases and its regulatory subunits, 13 strains revealed to have abnormal levels of lipid droplets, 10 of them having high lipid droplet content. Strains deleted for type I protein phosphatases and related regulators (ppz2, gac1, bni4, type 2A phosphatase and its related regulator (pph21 and sap185, type 2C protein phosphatases (ptc1, ptc4, ptc7 and dual phosphatases (pps1, msg5 were catalogued as high-lipid droplet content strains. Only reg1, a targeting subunit of the type 1 phosphatase Glc7p, and members of the nutrient-sensitive TOR pathway (sit4 and the regulatory subunit sap190 were catalogued as low-lipid droplet content strains, which were studied further. We show that Snf1, the homologue of the mammalian AMP-activated kinase, is constitutively phosphorylated (hyperactive in sit4 and sap190 strains leading to a reduction of acetyl-CoA carboxylase activity. In conclusion, our fast and highly sensitive method permitted us to catalogue protein phosphatases involved in the regulation of LD metabolism and present evidence indicating that the TOR pathway and the SNF1/AMPK pathway are connected through the Sit4p-Sap190p pair in the control of lipid droplet biogenesis.

  10. Dual Role of Ancient Ubiquitous Protein 1 (AUP1) in Lipid Droplet Accumulation and Endoplasmic Reticulum (ER) Protein Quality Control

    Science.gov (United States)

    Klemm, Elizabeth J.; Spooner, Eric; Ploegh, Hidde L.

    2011-01-01

    Quality control of endoplasmic reticulum proteins involves the identification and engagement of misfolded proteins, dislocation of the misfolded protein across the endoplasmic reticulum (ER) membrane, and ubiquitin-mediated targeting to the proteasome for degradation. Ancient ubiquitous protein 1 (AUP1) physically associates with the mammalian HRD1-SEL1L complex, and AUP1 depletion impairs degradation of misfolded ER proteins. One of the functions of AUP1 in ER quality control is to recruit the soluble E2 ubiquitin-conjugating enzyme UBE2G2. We further show that the CUE domain of AUP1 regulates polyubiquitylation and facilitates the interaction of AUP1 with the HRD1 complex and with dislocation substrates. AUP1 localizes both to the ER and to lipid droplets. The AUP1 expression level affects the abundance of cellular lipid droplets and as such represents the first protein with lipid droplet regulatory activity to be linked to ER quality control. These findings indicate a possible connection between ER protein quality control and lipid droplets. PMID:21857022

  11. Clearing the outer mitochondrial membrane from harmful proteins via lipid droplets

    Czech Academy of Sciences Publication Activity Database

    Bischof, J.; Salzmann, M.; Streubel, M.K.; Hašek, Jiří; Geltinger, F.; Duschl, J.; Bresgen, N.; Briza, P.; Hašková, Danuša; Lejsková, Renata; Sopjani, M.; Richter, K.; Rinnerthaler, M.

    2017-01-01

    Roč. 3, March 20 (2017), č. článku 17016. E-ISSN 2058-7716 R&D Projects: GA ČR(CZ) GA16-05497S; GA MŠk(CZ) 7AMB16AT006 Institutional support: RVO:61388971 Keywords : mitochondrial membrane * harmful protein s * lipid droplets Subject RIV: EE - Microbiology, Virology OBOR OECD: Microbiology

  12. Diacylglycerol Enrichment of Endoplasmic Reticulum or Lipid Droplets Recruits Perilipin 3/TIP47 during Lipid Storage and Mobilization*

    Science.gov (United States)

    Skinner, James R.; Shew, Trevor M.; Schwartz, Danielle M.; Tzekov, Anatoly; Lepus, Christin M.; Abumrad, Nada A.; Wolins, Nathan E.

    2009-01-01

    Fatty acid-induced triacylglycerol synthesis produces triacylglycerol droplets with a protein coat that includes perilipin 3/TIP47 and perilipin 4/S3-12. This study addresses the following two questions. Where do lipid droplets emerge, and how are their coat proteins recruited? We show that perilipin 3- and perilipin 4-coated lipid droplets emerge along the endoplasmic reticulum (ER). Blocking membrane trafficking with AlF4− during fatty acid-induced triacylglycerol synthesis drove perilipin 3 to the tubular ER. Forskolin, which like AlF4− activates adenylate cyclase, did not redistribute perilipin 3, but when added together with AlF4− perilipin 3 was recruited to lipid droplets rather than the ER. Thus inhibiting trafficking with AlF4− redistributed perilipin 3 differently under conditions of triacylglycerol synthesis (fatty acid addition) versus hydrolysis (forskolin) suggesting a shared acylglycerol-mediated mechanism. We tested whether diacylglycerol (DG), the immediate precursor of triacylglycerol and its first hydrolytic product, affects the distribution of perilipin 3. Stabilizing DG with the DG lipase inhibitor RHC80267 enhanced the perilipin 3 recruited to lipid droplets and raised DG levels in this fraction. Treating cells with a membrane-permeable DG recruited perilipin 3 to the ER. Stabilizing DG, by blocking its hydrolysis with RHC80267 or its acylation with triacsin C, enhanced recruitment of perilipin 3 to the ER. Expressing the ER enzyme DGAT1, which removes DG by converting it to triacylglycerol, attenuated perilipin 3 DG-induced ER recruitment. Membrane-permeable DG also drove perilipin 4 and 5 onto the ER. Together the data suggest that these lipid droplet proteins are recruited to DG-enriched membranes thereby linking lipid coat proteins to the metabolic state of the cell. PMID:19748893

  13. p53-inducible DHRS3 Is an Endoplasmic Reticulum Protein Associated with Lipid Droplet Accumulation*

    Science.gov (United States)

    Deisenroth, Chad; Itahana, Yoko; Tollini, Laura; Jin, Aiwen; Zhang, Yanping

    2011-01-01

    The transcription factor p53 plays a critical role in maintaining homeostasis as it relates to cellular growth, proliferation, and metabolism. In an effort to identify novel p53 target genes, a microarray approach was utilized to identify DHRS3 (also known as retSDR1) as a robust candidate gene. DHRS3 is a highly conserved member of the short chain alcohol dehydrogenase/reductase superfamily with a reported role in lipid and retinoid metabolism. Here, we demonstrate that DHRS3 is an endoplasmic reticulum (ER) protein that is shuttled to the ER via an N-terminal endoplasmic reticulum targeting signal. One important function of the ER is synthesis of neutral lipids that are packaged into lipid droplets whose biogenesis occurs from ER-derived membranes. DHRS3 is enriched at focal points of lipid droplet budding where it also localizes to the phospholipid monolayer of ER-derived lipid droplets. p53 promotes lipid droplet accumulation in a manner consistent with DHRS3 enrichment in the ER. As a p53 target gene, the observations of Dhrs3 location and potential function provide novel insight into an unexpected role for p53 in lipid droplet dynamics with implications in cancer cell metabolism and obesity. PMID:21659514

  14. Front-face fluorescence spectroscopy study of globular proteins in emulsions: influence of droplet flocculation.

    Science.gov (United States)

    Rampon, V; Genot, C; Riaublanc, A; Anton, M; Axelos, M A V; McClements, D J

    2003-04-23

    Measurement of the intensity (I(MAX)) and/or wavelength (lambda(MAX)) of the maximum in the tryptophan (TRP) emission spectrum using front-face fluorescence spectroscopy (FFFS) can be used to provide information about the molecular environment of proteins in nondiluted emulsions. Many protein-stabilized emulsions in the food industry are flocculated, and therefore, we examined the influence of droplet flocculation on FFFS. Stock oil-in-water emulsions stabilized by bovine serum albumin were prepared by high-pressure valve homogenization (30 wt % n-hexadecane, 0.35 wt % BSA, pH 7). These emulsions were used to create model systems with different degrees of droplet flocculation, either by changing the pH, adding surfactant, or adding xanthan. Emulsions (21 wt % n-hexadecane, 0.22 wt % BSA) with different pH (5 and 7) and molar ratios of Tween 20 to BSA (R = 0-131) were prepared by dilution of the stock emulsion. As the surfactant concentration was increased, the protein was displaced from the droplet surfaces, which caused an increase in both I(MAX) and lambda(MAX), because of the change in TRP environment. The dependence of I(MAX) and lambda(MAX) on surfactant concentration followed a similar pattern in emulsions that were initially flocculated (pH 5) and nonflocculated (pH 7). Relatively small changes in FFFS emission spectra were observed in emulsions (21 wt % n-hexadecane, 0.22 wt % BSA, pH 7) with different levels of depletion flocculation induced by adding xanthan. These results suggested that droplet flocculation did not have a major impact on FFFS. This study shows that FFFS is a powerful technique for nondestructively providing information about the molecular environment of proteins in concentrated and flocculated protein-stabilized emulsions. Nevertheless, in general the suitability of the technique may also depend on protein type and the nature of the physicochemical matrix surrounding the proteins.

  15. An ER Protein Functionally Couples Neutral Lipid Metabolism on Lipid Droplets to Membrane Lipid Synthesis in the ER

    DEFF Research Database (Denmark)

    Markgraf, Daniel F; Klemm, Robin W; Junker, Mirco

    2014-01-01

    Eukaryotic cells store neutral lipids such as triacylglycerol (TAG) in lipid droplets (LDs). Here, we have addressed how LDs are functionally linked to the endoplasmic reticulum (ER). We show that, in S. cerevisiae, LD growth is sustained by LD-localized enzymes. When LDs grow in early stationary...... phase, the diacylglycerol acyl-transferase Dga1p moves from the ER to LDs and is responsible for all TAG synthesis from diacylglycerol (DAG). During LD breakdown in early exponential phase, an ER membrane protein (Ice2p) facilitates TAG utilization for membrane-lipid synthesis. Ice2p has a cytosolic...... and explain how cells switch neutral lipid metabolism from storage to consumption....

  16. A random-access microarray for programmable droplet storage, retrieval and manipulation

    International Nuclear Information System (INIS)

    Tseng, Yi-Ming; Wang, Jhih-Jhe; Su, Yu-Chuan

    2012-01-01

    This article presents an integrated microfluidic system that is capable of programmably metering, entrapping, coalescing, addressably storing, retrieving and manipulating emulsion droplets. A multilayer, flexible PDMS chip with specially designed fluidic channels dynamically reconfigured by pneumatically actuated diaphragms is utilized to integrate a variety of droplet manipulation schemes. Once droplets are formed, their motions are coordinated by a 2D multiplexing scheme, which exploits the bidirectional movement of diaphragms to implement a random-access microarray. In the prototype demonstration, a PDMS molding and bonding process is used to fabricate the proposed microfluidic system. Emulsion droplets with desired volumes and compositions are produced, addressably stored, manipulated and retrieved from a 4 × 4 array, which employs just 4 (= 2 × log 2 4) control inputs for the operation. It has been demonstrated that (1) the integration of droplet manipulation and 2D multiplexing schemes can be achieved readily using bidirectional diaphragm valves, (2) multiplexing of an N × N array could be realized utilizing only 2 × log 2 N control inputs and (3) a multifunctional, random-access microarray can be accomplished employing a multilayer PDMS chip. As such, the demonstrated random-access microarray could potentially serve as a platform for continuous tracking and multistep processing of emulsion droplets, which is desired for various biological and chemical applications. (paper)

  17. Biomimetic materials for protein storage and transport

    Science.gov (United States)

    Firestone, Millicent A [Elmhurst, IL; Laible, Philip D [Villa Park, IL

    2012-05-01

    The invention provides a method for the insertion of protein in storage vehicles and the recovery of the proteins from the vehicles, the method comprising supplying isolated protein; mixing the isolated protein with a fluid so as to form a mixture, the fluid comprising saturated phospholipids, lipopolymers, and a surfactant; cycling the mixture between a first temperature and a second temperature; maintaining the mixture as a solid for an indefinite period of time; diluting the mixture in detergent buffer so as to disrupt the composition of the mixture, and diluting to disrupt the fluid in its low viscosity state for removal of the guest molecules by, for example, dialysis, filtering or chromatography dialyzing/filtering the emulsified solid.

  18. 17β-Hydroxysteroid dehydrogenase type 13 is a liver-specific lipid droplet-associated protein

    International Nuclear Information System (INIS)

    Horiguchi, Yuka; Araki, Makoto; Motojima, Kiyoto

    2008-01-01

    17β-Hydroxysteroid dehydrogenase (17βHSD) type 13 is identified as a new lipid droplet-associated protein. 17βHSD type 13 has an N-terminal sequence similar to that of 17βHSD type 11, and both sequences function as an endoplasmic reticulum and lipid droplet-targeting signal. Localization of native 17βHSD type 13 on the lipid droplets was confirmed by subcellular fractionation and Western blotting. In contrast to 17βHSD type 11, however, expression of 17βHSD type 13 is largely restricted to the liver and is not enhanced by peroxisome proliferator-activated receptor α and its ligand. Instead the expression level of 17βHSD type 13 in the receptor-null mice was increased several-fold. 17βHSD type 13 may have a distinct physiological role as a lipid droplet-associated protein in the liver

  19. An ER protein functionally couples neutral lipid metabolism on lipid droplets to membrane lipid synthesis in the ER.

    Science.gov (United States)

    Markgraf, Daniel F; Klemm, Robin W; Junker, Mirco; Hannibal-Bach, Hans K; Ejsing, Christer S; Rapoport, Tom A

    2014-01-16

    Eukaryotic cells store neutral lipids such as triacylglycerol (TAG) in lipid droplets (LDs). Here, we have addressed how LDs are functionally linked to the endoplasmic reticulum (ER). We show that, in S. cerevisiae, LD growth is sustained by LD-localized enzymes. When LDs grow in early stationary phase, the diacylglycerol acyl-transferase Dga1p moves from the ER to LDs and is responsible for all TAG synthesis from diacylglycerol (DAG). During LD breakdown in early exponential phase, an ER membrane protein (Ice2p) facilitates TAG utilization for membrane-lipid synthesis. Ice2p has a cytosolic domain with affinity for LDs and is required for the efficient utilization of LD-derived DAG in the ER. Ice2p breaks a futile cycle on LDs between TAG degradation and synthesis, promoting the rapid relocalization of Dga1p to the ER. Our results show that Ice2p functionally links LDs with the ER and explain how cells switch neutral lipid metabolism from storage to consumption. Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.

  20. An ER Protein Functionally Couples Neutral Lipid Metabolism on Lipid Droplets to Membrane Lipid Synthesis in the ER

    Directory of Open Access Journals (Sweden)

    Daniel F. Markgraf

    2014-01-01

    Full Text Available Eukaryotic cells store neutral lipids such as triacylglycerol (TAG in lipid droplets (LDs. Here, we have addressed how LDs are functionally linked to the endoplasmic reticulum (ER. We show that, in S. cerevisiae, LD growth is sustained by LD-localized enzymes. When LDs grow in early stationary phase, the diacylglycerol acyl-transferase Dga1p moves from the ER to LDs and is responsible for all TAG synthesis from diacylglycerol (DAG. During LD breakdown in early exponential phase, an ER membrane protein (Ice2p facilitates TAG utilization for membrane-lipid synthesis. Ice2p has a cytosolic domain with affinity for LDs and is required for the efficient utilization of LD-derived DAG in the ER. Ice2p breaks a futile cycle on LDs between TAG degradation and synthesis, promoting the rapid relocalization of Dga1p to the ER. Our results show that Ice2p functionally links LDs with the ER and explain how cells switch neutral lipid metabolism from storage to consumption.

  1. Engineering of soybean seed storage proteins

    International Nuclear Information System (INIS)

    Dickinson, C.D.; Floener, L.A.; Evans, R.P.; Nielsen, N.C.

    1987-01-01

    Protein engineering is one approach to the improvement of seed quality. With this in mind, a rapid in vitro system has been developed to assay the effect structural modifications have on the assembly of glycinin and β-conglycinin subunit complexes. Transcription plasmids were constructed for production of synthetic glycinin and β-conglycinin mRNAs by SP6 RNA-polymerase. Radiolabeled translation products from these messages were tested for their ability to form complexes. Gy4 and Gy5 proglycinins (group-2 subunits) and the a-subunit of β-conglycinin self-assembled into trimers. Proglycinin Gy2 (group-1 subunit) did not self-assemble, but assembled into mixed trimers in combination with Gy4 proglycinin. No assembly was observed for preproglycinins Gyl and Gy4, or for a Gy4 proglycinin which lacked 27 amino acids in a highly conserved internal sequence. Insertion of alternating MET-ARG residues in predicted turn regions of a hypervariable sequence in Gy4 proglycinin were tolerated when the string was short but inhibited trimer assembly as it became longer. The response to several different long deletions in this hypervariable region have also been tested. Different levels of trimer assembly were obtained and may depend on the secondary structures of the regions being joined in the engineered subunits. This system will be useful to study the assembly of storage protein complexes and to screen against modifications that interfere with subunit assembly

  2. Liver X receptors balance lipid stores in hepatic stellate cells through Rab18, a retinoid responsive lipid droplet protein.

    Science.gov (United States)

    O'Mahony, Fiona; Wroblewski, Kevin; O'Byrne, Sheila M; Jiang, Hongfeng; Clerkin, Kara; Benhammou, Jihane; Blaner, William S; Beaven, Simon W

    2015-08-01

    Liver X receptors (LXRs) are determinants of hepatic stellate cell (HSC) activation and liver fibrosis. Freshly isolated HSCs from Lxrαβ(-/-) mice have increased lipid droplet (LD) size, but the functional consequences of this are unknown. Our aim was to determine whether LXRs link cholesterol to retinoid storage in HSCs and how this impacts activation. Primary HSCs from Lxrαβ(-/-) and wild-type mice were profiled by gene array during in vitro activation. Lipid content was quantified by high-performance liquid chromatography and mass spectroscopy. Primary HSCs were treated with nuclear receptor ligands, transfected with small interfering RNA and plasmid constructs, and analyzed by immunocytochemistry. Lxrαβ(-/-) HSCs have increased cholesterol and retinyl esters. The retinoid increase drives intrinsic retinoic acid receptor signaling, and activation occurs more rapidly in Lxrαβ(-/-) HSCs. We identify Rab18 as a novel retinoic acid-responsive, LD-associated protein that helps mediate stellate cell activation. Rab18 mRNA, protein, and membrane insertion increase during activation. Both Rab18 guanosine triphosphatase activity and isoprenylation are required for stellate cell LD loss and induction of activation markers. These phenomena are accelerated in Lxrαβ(-/-) HSCs, where there is greater retinoic acid flux. Conversely, Rab18 knockdown retards LD loss in culture and blocks activation, just like the functional mutants. Rab18 is also induced with acute liver injury in vivo. Retinoid and cholesterol metabolism are linked in stellate cells by the LD-associated protein Rab18. Retinoid overload helps explain the profibrotic phenotype of Lxrαβ(-/-) mice, and we establish a pivotal role for Rab18 GTPase activity and membrane insertion in wild-type stellate cell activation. Interference with Rab18 may have significant therapeutic benefit in ameliorating liver fibrosis. © 2015 by the American Association for the Study of Liver Diseases.

  3. Using droplet-on-demand based printing to guide self-assembly in a peptide-protein based bioink

    Science.gov (United States)

    Hedegaard, Clara; Collin, Estelle; Redondo-Gomez, Carlos; Nguyen, Luong T. H.; Ng, Kee Woei; Castrejon-Pita, Alfonso A.; Castrejon-Pita, J. Rafael; Mata, Alvaro

    2017-11-01

    Tissue engineering aims to capture details of the extracellular matrix (ECM) that stimulate tissue regeneration. Advanced biofabrication techniques have enabled structural complexity, however they are restricted by the choice of material due to stringent printing requirements, leading to a lack of nanoscale control and molecular versatility. In this project, we exploit the dynamics of droplet fluid interactions combined with the co-assembly of peptide amphiphiles (PAs) with biomolecules/proteins to develop a new approach to droplet-based biofabrication. A custom-made droplet generator was developed and used to controllably dispense droplets of PA into a protein solution resulting in gel formation within milliseconds. Taking advantage of the interfacial and inertial forces during the droplet/liquid interaction, it is possible to control the co-assembly kinetics, to give rise to aligned or disordered nanofibers, hydrogel structures of different geometries and sizes, surface topographies, and higher-ordered structures made from multiple hydrogels. The process allows multiple cell types to be spatially distributed on the outside or embedded within the ECM mimetic scaffolds, whilst exhibiting high cell viability (>88%). ERC Starting Grant (STROFUNSCAFF), FP7-PEOPLE-2013-CIG Biomorph and the Royal Society.

  4. Fat-specific protein 27 regulates storage of triacylglycerol

    DEFF Research Database (Denmark)

    Keller, P.; Petrie, J.T.; Rose, P. De

    2008-01-01

    FSP27 (fat-specific protein 27) is a member of the cell death-inducing DNA fragmentation factor-alpha-like effector (CIDE) family. Although Cidea and Cideb were initially characterized as activators of apoptosis, recent studies have demonstrated important metabolic roles for these proteins...... in several cell types without induction of adipocyte genes. Increased triacylglycerol is likely due to decreased beta-oxidation of nonesterified fatty acids. Altered flux of fatty acids into triacylglycerol may be a direct effect of FSP27 function, which is localized to lipid droplets in 293T cells and 3T3-L...... decreases with total fat mass but is not associated with measures of insulin resistance (e.g. homeostasis model assessment). Together, these data indicate that FSP27 binds to lipid droplets and regulates their enlargement Udgivelsesdato: 2008/5/23...

  5. Cross-linking proteins by laccase: Effects on the droplet size and rheology of emulsions stabilized by sodium caseinate.

    Science.gov (United States)

    Sato, A C K; Perrechil, F A; Costa, A A S; Santana, R C; Cunha, R L

    2015-09-01

    The aim of this work was to evaluate the influence of laccase and ferulic acid on the characteristics of oil-in-water emulsions stabilized by sodium caseinate at different pH (3, 5 and 7). Emulsions were prepared by high pressure homogenization of soybean oil with sodium caseinate solution containing varied concentrations of laccase (0, 1 and 5mg/mL) and ferulic acid (5 and 10mM). Laccase treatment and pH exerted a strong influence on the properties with a consequent effect on stability, structure and rheology of emulsions stabilized by Na-caseinate. At pH7, O/W emulsions were kinetically stable due to the negative protein charge which enabled electrostatic repulsion between oil droplets resulting in an emulsion with small droplet size, low viscosity, pseudoplasticity and viscoelastic properties. The laccase treatment led to emulsions showing shear-thinning behavior as a result of a more structured system. O/W emulsions at pH5 and 3 showed phase separation due to the proximity to protein pI, but the laccase treatment improved their stability of emulsions especially at pH3. At pH3, the addition of ferulic acid and laccase produced emulsions with larger droplet size but with narrower droplet size distribution, increased viscosity, pseudoplasticity and viscoelastic properties (gel-like behavior). Comparing laccase treatments, the combined addition of laccase and ferulic acid generally produced emulsions with lower stability (pH5), larger droplet size (pH3, 5 and 7) and higher pseudoplasticity (pH5 and 7) than emulsion with only ferulic acid. The results suggested that the cross-linking of proteins by laccase and ferulic acid improved protein emulsifying properties by changing functional mechanisms of the protein on emulsion structure and rheology, showing that sodium caseinate can be successfully used in acid products when treated with laccase. Copyright © 2015 Elsevier Ltd. All rights reserved.

  6. Bidirectional lipid droplet velocities are controlled by differential binding strengths of HCV core DII protein.

    Directory of Open Access Journals (Sweden)

    Rodney K Lyn

    Full Text Available Host cell lipid droplets (LD are essential in the hepatitis C virus (HCV life cycle and are targeted by the viral capsid core protein. Core-coated LDs accumulate in the perinuclear region and facilitate viral particle assembly, but it is unclear how mobility of these LDs is directed by core. Herein we used two-photon fluorescence, differential interference contrast imaging, and coherent anti-Stokes Raman scattering microscopies, to reveal novel core-mediated changes to LD dynamics. Expression of core protein's lipid binding domain II (DII-core induced slower LD speeds, but did not affect directionality of movement on microtubules. Modulating the LD binding strength of DII-core further impacted LD mobility, revealing the temporal effects of LD-bound DII-core. These results for DII-core coated LDs support a model for core-mediated LD localization that involves core slowing down the rate of movement of LDs until localization at the perinuclear region is accomplished where LD movement ceases. The guided localization of LDs by HCV core protein not only is essential to the viral life cycle but also poses an interesting target for the development of antiviral strategies against HCV.

  7. Protein construct storage: Bayesian variable selection and prediction with mixtures.

    Science.gov (United States)

    Clyde, M A; Parmigiani, G

    1998-07-01

    Determining optimal conditions for protein storage while maintaining a high level of protein activity is an important question in pharmaceutical research. A designed experiment based on a space-filling design was conducted to understand the effects of factors affecting protein storage and to establish optimal storage conditions. Different model-selection strategies to identify important factors may lead to very different answers about optimal conditions. Uncertainty about which factors are important, or model uncertainty, can be a critical issue in decision-making. We use Bayesian variable selection methods for linear models to identify important variables in the protein storage data, while accounting for model uncertainty. We also use the Bayesian framework to build predictions based on a large family of models, rather than an individual model, and to evaluate the probability that certain candidate storage conditions are optimal.

  8. Arabidopsis lipid droplet-associated protein (LDAP)–interacting protein (LDIP) influences lipid droplet size and neutral lipid homeostasis in both leaves and seeds

    Science.gov (United States)

    Cytoplasmic lipid droplets (LDs) are found in all types of plant cells where they are derived from the endoplasmic reticulum and function as a repository for neutral lipids, as well as serving in lipid remodelling and signalling. However, the mechanisms underlying the formation and functioning of pl...

  9. The role of organelle proteins in carotenoid droplet dispersion in goldfish xanthophore

    International Nuclear Information System (INIS)

    Wu, B.Y.

    1988-01-01

    In this study, I report on the evidence for the possible role of two carotenoid droplet (CD) proteins in CD dispersion. In order to identify the target of the ATP analogue 8-N 3 ATP, CDs, one of the major components in permeabilized xanthophore residues, were isolated by sucrose density gradient centrifugation and were tested for ATPase activity. An ATPase activity was found associated with CDs. Polypeptides analysis of the two subpopulations of 3 H-leucine labeled CDs by one dimensional and two dimensional polyacrylamide gel electrophoresis showed that there is more actin associated with HCD than with LCD, suggesting that the CD-ATPase may be an apparent actin activated ATPase. The properties of the CD-ATPase have been partially characterized with regard to its ionic requirement and the effects of known ATPase inhibitors and potential activators. Monoclonal antibodies were generated against CD proteins. The major monoclonal antibodies obtained are anti-p57s. The effects of these anti-p57s on dispersion of CDs in permeabilized xanthophores were determined and it was found that some inhibit CD dispersion while others do not

  10. The perilipin homologue, lipid storage droplet 2, regulates sleep homeostasis and prevents learning impairments following sleep loss.

    Directory of Open Access Journals (Sweden)

    Matthew S Thimgan

    2010-08-01

    Full Text Available Extended periods of waking result in physiological impairments in humans, rats, and flies. Sleep homeostasis, the increase in sleep observed following sleep loss, is believed to counter the negative effects of prolonged waking by restoring vital biological processes that are degraded during sleep deprivation. Sleep homeostasis, as with other behaviors, is influenced by both genes and environment. We report here that during periods of starvation, flies remain spontaneously awake but, in contrast to sleep deprivation, do not accrue any of the negative consequences of prolonged waking. Specifically, the homeostatic response and learning impairments that are a characteristic of sleep loss are not observed following prolonged waking induced by starvation. Recently, two genes, brummer (bmm and Lipid storage droplet 2 (Lsd2, have been shown to modulate the response to starvation. bmm mutants have excess fat and are resistant to starvation, whereas Lsd2 mutants are lean and sensitive to starvation. Thus, we hypothesized that bmm and Lsd2 may play a role in sleep regulation. Indeed, bmm mutant flies display a large homeostatic response following sleep deprivation. In contrast, Lsd2 mutant flies, which phenocopy aspects of starvation as measured by low triglyceride stores, do not exhibit a homeostatic response following sleep loss. Importantly, Lsd2 mutant flies are not learning impaired after sleep deprivation. These results provide the first genetic evidence, to our knowledge, that lipid metabolism plays an important role in regulating the homeostatic response and can protect against neuronal impairments induced by prolonged waking.

  11. In situ X-ray scattering studies of protein solution droplets drying on micro- and nanopatterned superhydrophobic PMMA surfaces.

    Science.gov (United States)

    Accardo, Angelo; Gentile, Francesco; Mecarini, Federico; De Angelis, Francesco; Burghammer, Manfred; Di Fabrizio, Enzo; Riekel, Christian

    2010-09-21

    Superhydrophobic poly(methyl methacrylate) surfaces with contact angles of ∼170° and high optical and X-ray transparencies have been fabricated through the use of optical lithography and plasma etching. The surfaces contain either a microscale pattern of micropillars or a random nanofibrillar pattern. Nanoscale asperities on top of the micropillars closely resemble Nelumbo nucifera lotus leaves. The evolution of the contact angle of water and lysozyme solution droplets during evaporation was studied on the micro- and nanopatterned surfaces, showing in particular contact-line pinning for the protein solution droplet on the nanopatterned surface. The microstructural evolution of lysozyme solution droplets was studied on both types of surfaces in situ under nearly contact-free conditions by synchrotron radiation microbeam wide-angle and small-angle X-ray scattering revealing the increasing protein concentration and the onset of precipitation. The solid residuals show hollow sphere morphologies. Rastermicrodiffraction of the detached residuals suggests about a 1/3 volume fraction of ≥17 nm lysozyme nanocrystalline domains and about a 2/3 short-range-order volume fraction. About 5-fold larger nanocrystalline domains were observed at the attachment points of the sphere to the substrates, which is attributed to particle growth in a shear flow. Such surfaces represent nearly contact-free sample supports for studies of inorganic and organic solution droplets, which find applications in biochips.

  12. Subcellular localization of skeletal muscle lipid droplets and PLIN family proteins OXPAT and ADRP at rest and following contraction in rat soleus muscle.

    Science.gov (United States)

    MacPherson, Rebecca E K; Herbst, Eric A F; Reynolds, Erica J; Vandenboom, Rene; Roy, Brian D; Peters, Sandra J

    2012-01-01

    Skeletal muscle lipid droplet-associated proteins (PLINs) are thought to regulate lipolysis through protein-protein interactions on the lipid droplet surface. In adipocytes, PLIN2 [adipocyte differentiation-related protein (ADRP)] is found only on lipid droplets, while PLIN5 (OXPAT, expressed only in oxidative tissues) is found both on and off the lipid droplet and may be recruited to lipid droplet membranes when needed. Our purpose was to determine whether PLIN5 is recruited to lipid droplets with contraction and to investigate the myocellular location and colocalization of lipid droplets, PLIN2, and PLIN5. Rat solei were isolated, and following a 30-min equilibration period, they were assigned to one of two groups: 1) 30 min of resting incubation and 2) 30 min of stimulation (n = 10 each). Immunofluorescence microscopy was used to determine subcellular content, distribution, and colocalization of lipid droplets, PLIN2, and PLIN5. There was a main effect for lower lipid and PLIN2 content in stimulated compared with rested muscles (P muscles (P = 0.001, r(2) = 0.99) and linearly in stimulated muscles (slope = -0.0023 ± 0.0006, P muscles (P contraction in isolated skeletal muscle.

  13. Isolation of Protein Storage Vacuoles and Their Membranes.

    Science.gov (United States)

    Shimada, Tomoo; Hara-Nishimura, Ikuko

    2017-01-01

    Protein-storage vacuoles (PSVs) are specialized vacuoles that sequester large amounts of storage proteins. During seed development, PSVs are formed de novo and/or from preexisting lytic vacuoles. Seed PSVs can be subdivided into four distinct compartments: membrane, globoid, matrix, and crystalloid. In this chapter, we introduce easy methods for isolation of PSVs and their membranes from pumpkin seeds. These methods facilitate the identification and characterization of PSV components.

  14. Lipid droplet-associated proteins in high-fat fed mice with the effects of voluntary running and diet change.

    Science.gov (United States)

    Rinnankoski-Tuikka, Rita; Hulmi, Juha J; Torvinen, Sira; Silvennoinen, Mika; Lehti, Maarit; Kivelä, Riikka; Reunanen, Hilkka; Kujala, Urho M; Kainulainen, Heikki

    2014-08-01

    The relation between lipid accumulation and influence of exercise on insulin sensitivity is not straightforward. A proper balance between lipid droplet synthesis, lipolysis, and oxidative metabolism would ensure low local intramyocellular fatty acid levels, thereby possibly protecting against lipotoxicity-associated insulin resistance. This study investigated whether the accumulation of triglycerides and lipid droplets in response to high availability of fatty acids after high-fat feeding would parallel the abundance of intramyocellular perilipin proteins, especially PLIN5. The effects on these variables after diet change or voluntary running exercise intervention in skeletal muscle were also investigated. During a 19-week experiment, C57BL/6J mice were studied in six different groups: low-fat diet sedentary, low-fat diet active, high-fat diet sedentary, high-fat diet active and two groups which were high-fat sedentary for nine weeks, after which divided into low-fat sedentary or low-fat active groups. Myocellular triglyceride concentration and perilipin protein expression levels were assessed. We show that, concurrently with impaired insulin sensitivity, the expression level of PLIN5 and muscular triglyceride concentration increased dramatically after high-fat diet. These adaptations were reversible after the diet change intervention with no additional effect of exercise. After high-fat diet, lipid droplets become larger providing more surface area for PLIN5. We suggest that PLIN5 is an important regulator of lipid droplet turnover in altered conditions of fatty acid supply and consumption. Imbalances in lipid droplet metabolism and turnover might lead to lipotoxicity-related insulin resistance. Copyright © 2014 Elsevier Inc. All rights reserved.

  15. [Dynamics of Irreversible Evaporation of a Water-Protein Droplet and a Problem of Structural and Dynamical Experiments with Single Molecules].

    Science.gov (United States)

    Shaitan, K V; Armeev, G A; Shaytan, A K

    2016-01-01

    We discuss the effect of isothermal and adiabatic evaporation of water on the state of a water-protein droplet. The discussed problem is of current importance due to development of techniques to perform single molecule experiments using free electron lasers. In such structure-dynamic experiments the delivery of a sample into the X-ray beam is performed using the microdroplet injector. The time between the injection and delivery is in the order of microseconds. In this paper we developed a specialized variant of all-atom molecular dynamics simulations for the study of irreversible isothermal evaporation of the droplet. Using in silico experiments we determined the parameters of isothermal evaporation of the water-protein droplet with the sodium and chloride ions in the concentration range of 0.3 M at different temperatures. The energy of irreversible evaporation determined from in silico experiments at the initial stages of evaporation virtually coincides with the specific heat of evaporation for water. For the kinetics of irreversible adiabatic evaporation an exact analytical solution was obtained in the limit of high thermal conductivity of the droplet (or up to the droplet size of -100 Å). This analytical solution incorporates parameters that are determined using in silico. experiments on isothermal droplet evaporation. We show that the kinetics of adiabatic evaporation and cooling of the droplet scales with the droplet size. Our estimates of the water-protemi droplet. freezing rate in the adiabatic regime in a vacuum chamber show that additional techniques for stabilizing the temperature inside the droplet should be used in order to study the conformational transitions of the protein in single molecules. Isothermal and quasi-isothermal conditions are most suitable for studying the conformational transitions upon object functioning. However, in this case it is necessary to take into account the effects of dehydration and rapid increase of ionic strength in an

  16. Behavior of whey protein concentrates under extreme storage conditions

    Science.gov (United States)

    The overseas demand for whey protein concentrates (WPC) has increased steadily in recent years. Emergency aid foods often include WPC, but shelf-life studies of whey proteins under different shipment and storage conditions have not been conducted in the last 50 yr. Microbial quality, compound form...

  17. Seed storage protein components are associated with curled ...

    African Journals Online (AJOL)

    PRECIOUS

    2009-11-16

    Nov 16, 2009 ... analysis suggests that the two increased protein spots in mutants were ... The main objective of this work was to gain further understanding of the influence of curled cotyledon on the seed storage protein components in soybean by com- .... cotyledon formation during Arabidopsis embryogenesis: interaction.

  18. SLDP: a novel protein related to caleosin is associated with the endosymbiotic Symbiodinium lipid droplets from Euphyllia glabrescens.

    Science.gov (United States)

    Pasaribu, Buntora; Lin, I-Ping; Tzen, Jason T C; Jauh, Guang-Yuh; Fan, Tung-Yung; Ju, Yu-Min; Cheng, Jing-O; Chen, Chii-Shiarng; Jiang, Pei-Luen

    2014-10-01

    Intracellular lipid droplets (LDs) have been proposed to play a key role in the mutualistic endosymbiosis between reef-building corals and the dinoflagellate endosymbiont Symbiodinium spp. This study investigates and identifies LD proteins in Symbiodinium from Euphyllia glabrescens. Discontinuous Percoll gradient centrifugation was used to separate Symbiodinium cells from E. glabrescens tentacles. Furthermore, staining with a fluorescent probe, Nile red, indicated that lipids accumulated in that freshly isolated Symbiodinium cells and lipid analyses further showed polyunsaturated fatty acids (PUFA) was abundant. The stable LDs were purified from endosymbiotic Symbiodinium cells. The structural integrity of the Symbiodinium LDs was maintained via electronegative repulsion and steric hindrance possibly provided by their surface proteins. Protein extracts from the purified LDs revealed a major protein band with a molecular weight of 20 kDa, which was termed Symbiodinium lipid droplet protein (SLDP). Interestingly, immunological cross-recognition analysis revealed that SLDP was detected strongly by the anti-sesame and anti-cycad caleosin antibodies. It was suggested that the stable Symbiodinium LDs were sheltered by this unique structural protein and was suggested that SLDP might be homologous to caleosin to a certain extent.

  19. Differential solvation of intrinsically disordered linkers drives the formation of spatially organized droplets in ternary systems of linear multivalent proteins

    Science.gov (United States)

    Harmon, Tyler S.; Holehouse, Alex S.; Pappu, Rohit V.

    2018-04-01

    Intracellular biomolecular condensates are membraneless organelles that encompass large numbers of multivalent protein and nucleic acid molecules. The bodies assemble via a combination of liquid–liquid phase separation and gelation. A majority of condensates included multiple components and show multilayered organization as opposed to being well-mixed unitary liquids. Here, we put forward a simple thermodynamic framework to describe the emergence of spatially organized droplets in multicomponent systems comprising of linear multivalent polymers also known as associative polymers. These polymers, which mimic proteins and/or RNA have the architecture of domains or motifs known as stickers that are interspersed by flexible spacers known as linkers. Using a minimalist numerical model for a four-component system, we have identified features of linear multivalent molecules that are necessary and sufficient for generating spatially organized droplets. We show that differences in sequence-specific effective solvation volumes of disordered linkers between interaction domains enable the formation of spatially organized droplets. Molecules with linkers that are preferentially solvated are driven to the interface with the bulk solvent, whereas molecules that have linkers with negligible effective solvation volumes form cores in the core–shell architectures that emerge in the minimalist four-component systems. Our modeling has relevance for understanding the physical determinants of spatially organized membraneless organelles.

  20. pH dependence of the kinetics of interfacial tension changes during protein adsorption from sessile droplets on FEP-Teflon

    NARCIS (Netherlands)

    VanderVegt, W; Norde, W; VanderMei, HC; Busscher, HJ

    Interfacial tension changes during protein adsorption at both the solid-liquid and the liquid-vapor interface were measured simultaneously by ADSA-P from sessile droplets of protein solutions on fluoroethylenepropylene-Teflon. Four globular proteins of similar size, viz. lysozyme, ribonuclease,

  1. Corn Storage Protein - A Molecular Genetic Model

    Energy Technology Data Exchange (ETDEWEB)

    Messing, Joachim [Rutgers Univ., Piscataway, NJ (United States)

    2013-05-31

    Corn is the highest yielding crop on earth and probably the most valuable agricultural product of the United States. Because it converts sun energy through photosynthesis into starch and proteins, we addressed energy savings by focusing on protein quality. People and animals require essential amino acids derived from the digestion of proteins. If proteins are relatively low in certain essential amino acids, the crop becomes nutritionally defective and has to be supplemented. Such deficiency affects meat and fish production and countries where corn is a staple. Because corn seed proteins have relatively low levels of lysine and methionine, a diet has to be supplemented with soybeans for the missing lysine and with chemically synthesized methionine. We therefore have studied genes expressed during maize seed development and their chromosomal organization. A critical technical requirement for the understanding of the molecular structure of genes and their positional information was DNA sequencing. Because of the length of sequences, DNA sequencing methods themselves were insufficient for this type of analysis. We therefore developed the so-called “DNA shotgun sequencing” strategy, where overlapping DNA fragments were sequenced in parallel and used to reconstruct large DNA molecules via overlaps. Our publications became the most frequently cited ones during the decade of 1981-1990 and former Associate Director of Science for the Office of Basic Energy Sciences Patricia M. Dehmer presented our work as one of the great successes of this program. A major component of the sequencing strategy was the development of bacterial strains and vectors, which were also used to develop the first biotechnology crops. These crops possessed new traits thanks to the expression of foreign genes in plants. To enable such expression, chimeric genes had to be constructed using our materials and methods by the industry. Because we made our materials and methods freely available to

  2. Whey protein concentrate storage at elevated temperature and humidity

    Science.gov (United States)

    Dairy processors are finding new export markets for whey protein concentrate (WPC), a byproduct of cheesemaking, but they need to know if full-sized bags of this powder will withstand high temperature and relative humidity (RH) levels during unrefrigerated storage under tropical conditions. To answ...

  3. Bioprinting: Functional droplet networks

    Science.gov (United States)

    Durmus, Naside Gozde; Tasoglu, Savas; Demirci, Utkan

    2013-06-01

    Tissue-mimicking printed networks of droplets separated by lipid bilayers that can be functionalized with membrane proteins are able to spontaneously fold and transmit electrical currents along predefined paths.

  4. Plant storage proteins – the main nourisching products – from biosynthesis to cellular storage depots

    Directory of Open Access Journals (Sweden)

    Agnieszka Chmielnicka

    2017-06-01

    Full Text Available Storage proteins of legumes are one of the main components of the human and animal diet. The substances collected in their seeds have the pro-health values, supporting the prevention of many civilization diseases. However, there are still many uncertainties about the mechanisms leading to the production of nutritious seeds. It is also difficult to identify which of their constituents and in what final form are responsible for the observed protective effects in vivo. In this work, on the background of different types of storage proteins, these deposited mainly in legumes were in the focus of interest. They were characterized on the example of pea (Pisum sativum proteins. Mechanisms associated with their biosynthesis and transport to specific cellular compartments was presented. Ways of their post-translational processing, segregation and storage in the specific vacuoles were also discussed. Therefore, the paper presents the state-of-the-art knowledge concerning the processes making the accumulated protein deposits ready to use by plants, animals and humans.

  5. Interfacial Tension and Surface Pressure of High Density Lipoprotein, Low Density Lipoprotein, and Related Lipid Droplets

    DEFF Research Database (Denmark)

    Ollila, O. H. S.; Lamberg, A.; Lehtivaara, M.

    2012-01-01

    ) are essentially lipid droplets surrounded by specific proteins, their main function being to transport cholesterol. Interfacial tension and surface pressure of these particles are of great interest because they are related to the shape and the stability of the droplets and to protein adsorption at the interface....... Here we use coarse-grained molecular-dynamics simulations to consider a number of related issues by calculating the interfacial tension in protein-free lipid droplets, and in HDL and LDL particles mimicking physiological conditions. First, our results suggest that the curvature dependence......Lipid droplets play a central role in energy storage and metabolism on a cellular scale. Their core is comprised of hydrophobic lipids covered by a surface region consisting of amphiphilic lipids and proteins. For example, high and low density lipoproteins (HDL and LDL, respectively...

  6. ATGL and CGI-58 are lipid droplet proteins of the hepatic stellate cell line HSC-T6.

    Science.gov (United States)

    Eichmann, Thomas O; Grumet, Lukas; Taschler, Ulrike; Hartler, Jürgen; Heier, Christoph; Woblistin, Aaron; Pajed, Laura; Kollroser, Manfred; Rechberger, Gerald; Thallinger, Gerhard G; Zechner, Rudolf; Haemmerle, Günter; Zimmermann, Robert; Lass, Achim

    2015-10-01

    Lipid droplets (LDs) of hepatic stellate cells (HSCs) contain large amounts of vitamin A [in the form of retinyl esters (REs)] as well as other neutral lipids such as TGs. During times of insufficient vitamin A availability, RE stores are mobilized to ensure a constant supply to the body. To date, little is known about the enzymes responsible for the hydrolysis of neutral lipid esters, in particular of REs, in HSCs. In this study, we aimed to identify LD-associated neutral lipid hydrolases by a proteomic approach using the rat stellate cell line HSC-T6. First, we loaded cells with retinol and FAs to promote lipid synthesis and deposition within LDs. Then, LDs were isolated and lipid composition and the LD proteome were analyzed. Among other proteins, we found perilipin 2, adipose TG lipase (ATGL), and comparative gene identification-58 (CGI-58), known and established LD proteins. Bioinformatic search of the LD proteome for α/β-hydrolase fold-containing proteins revealed no yet uncharacterized neutral lipid hydrolases. In in vitro activity assays, we show that rat (r)ATGL, coactivated by rat (r)CGI-58, efficiently hydrolyzes TGs and REs. These findings suggest that rATGL and rCGI-58 are LD-resident proteins in HSCs and participate in the mobilization of both REs and TGs. Copyright © 2015 by the American Society for Biochemistry and Molecular Biology, Inc.

  7. Characterization of seed storage protein patterns of Heliotropium digynum

    Directory of Open Access Journals (Sweden)

    Mona Soliman Alwhibi

    2017-09-01

    Full Text Available Heliotropium digynum, is a shrub that has ecological importance. The height of the plant differs from one population to another and the difference in length of the inflorescence can be attributed to environmental factors, such as rainfall or type of soil and temperature. To date, no study has shed light on estimation in seed samples of H. digynum in Saudi Arabia. So, the aim is to evaluate and characterize the protein patterns of seed storage proteins of H. digynum to be used as fingerprint of this plant in Saudi Arabia. It is collected from different locations in the central region of Saudi Arabia and total protein extraction from plant was compared in SDS-PAGE. The genetic relationships among all cultivars were analyzed using UPGMA and NJ using Total Lab TL and in the same way using Jaccard Similarity Coefficient dendrogram using STATISTICA (ver.8 software. Results, our data show that amounts of protein are different, although they are of the same type or from the same geographical region. Amounts ranged between 22 and 1.5 mg/g of dry weight. Less amount of protein was obtained from the group of samples collected from Dir’iyah area, and the highest amount of protein was from the group of samples collected from Dyrab area in general.

  8. Characterization of seed storage protein patterns of Heliotropium digynum.

    Science.gov (United States)

    Alwhibi, Mona Soliman

    2017-09-01

    Heliotropium digynum , is a shrub that has ecological importance. The height of the plant differs from one population to another and the difference in length of the inflorescence can be attributed to environmental factors, such as rainfall or type of soil and temperature. To date, no study has shed light on estimation in seed samples of H. digynum in Saudi Arabia. So, the aim is to evaluate and characterize the protein patterns of seed storage proteins of H. digynum to be used as fingerprint of this plant in Saudi Arabia. It is collected from different locations in the central region of Saudi Arabia and total protein extraction from plant was compared in SDS-PAGE. The genetic relationships among all cultivars were analyzed using UPGMA and NJ using Total Lab TL and in the same way using Jaccard Similarity Coefficient dendrogram using STATISTICA (ver.8) software. Results, our data show that amounts of protein are different, although they are of the same type or from the same geographical region. Amounts ranged between 22 and 1.5 mg/g of dry weight. Less amount of protein was obtained from the group of samples collected from Dir'iyah area, and the highest amount of protein was from the group of samples collected from Dyrab area in general.

  9. Fat storage-inducing transmembrane (FIT or FITM proteins are related to lipid phosphatase/phosphotransferase enzymes

    Directory of Open Access Journals (Sweden)

    Matthew J Hayes

    2017-12-01

    Full Text Available Fat storage-inducing transmembrane (FIT or FITM proteins have been implicated in the partitioning of triacylglycerol to lipid droplets and the budding of lipid droplets from the ER. At the molecular level, the sole relevant interaction is that FITMs directly bind to triacyglycerol and diacylglycerol, but how they function at the molecular level is not known. Saccharomyces cerevisiae has two FITM homologues: Scs3p and Yft2p. Scs3p was initially identified because deletion leads to inositol auxotrophy, with an unusual sensitivity to addition of choline. This strongly suggests a role for Scs3p in phospholipid biosynthesis. Looking at the FITM family as widely as possible, we found that FITMs are widespread throughout eukaryotes, indicating presence in the last eukaryotic common ancestor. Protein alignments also showed that FITM sequences contain the active site of lipid phosphatase/phosphotransferase (LPT enzymes. This large family transfers phosphate-containing headgroups either between lipids or in exchange for water. We confirmed the prediction that FITMs are related to LPTs by showing that single amino-acid substitutions in the presumptive catalytic site prevented their ability to rescue growth of the mutants on low inositol/high choline media when over-expressed. The substitutions also prevented rescue of other phenotypes associated with loss of FITM in yeast, including mistargeting of Opi1p, defective ER morphology, and aberrant lipid droplet budding. These results suggest that Scs3p, Yft2p and FITMs in general are LPT enzymes involved in an as yet unknown critical step in phospholipid metabolism.

  10. Abseq: Ultrahigh-throughput single cell protein profiling with droplet microfluidic barcoding

    Science.gov (United States)

    Shahi, Payam; Kim, Samuel C.; Haliburton, John R.; Gartner, Zev J.; Abate, Adam R.

    2017-03-01

    Proteins are the primary effectors of cellular function, including cellular metabolism, structural dynamics, and information processing. However, quantitative characterization of proteins at the single-cell level is challenging due to the tiny amount of protein available. Here, we present Abseq, a method to detect and quantitate proteins in single cells at ultrahigh throughput. Like flow and mass cytometry, Abseq uses specific antibodies to detect epitopes of interest; however, unlike these methods, antibodies are labeled with sequence tags that can be read out with microfluidic barcoding and DNA sequencing. We demonstrate this novel approach by characterizing surface proteins of different cell types at the single-cell level and distinguishing between the cells by their protein expression profiles. DNA-tagged antibodies provide multiple advantages for profiling proteins in single cells, including the ability to amplify low-abundance tags to make them detectable with sequencing, to use molecular indices for quantitative results, and essentially limitless multiplexing.

  11. Dual Role of Ancient Ubiquitous Protein 1 (AUP1) in Lipid Droplet Accumulation and Endoplasmic Reticulum (ER) Protein Quality Control

    OpenAIRE

    Klemm, Elizabeth J.; Spooner, Eric; Ploegh, Hidde L.

    2011-01-01

    Quality control of endoplasmic reticulum proteins involves the identification and engagement of misfolded proteins, dislocation of the misfolded protein across the endoplasmic reticulum (ER) membrane, and ubiquitin-mediated targeting to the proteasome for degradation. Ancient ubiquitous protein 1 (AUP1) physically associates with the mammalian HRD1-SEL1L complex, and AUP1 depletion impairs degradation of misfolded ER proteins. One of the functions of AUP1 in ER quality control is to recruit t...

  12. Postprandial triglyceride-rich lipoproteins regulate perilipin-2 and perilipin-3 lipid-droplet-associated proteins in macrophages.

    Science.gov (United States)

    Varela, Lourdes M; López, Sergio; Ortega-Gómez, Almudena; Bermúdez, Beatriz; Buers, Insa; Robenek, Horst; Muriana, Francisco J G; Abia, Rocío

    2015-04-01

    Lipid accumulation in macrophages contributes to atherosclerosis. Within macrophages, lipids are stored in lipid droplets (LDs); perilipin-2 and perilipin-3 are the main LD-associated proteins. Postprandial triglyceride (TG)-rich lipoproteins induce LD accumulation in macrophages. The role of postprandial lipoproteins in perilipin-2 and perilipin-3 regulation was studied. TG-rich lipoproteins (TRLs) induced the levels of intracellular TGs, LDs and perilipin-2 protein expression in THP-1 macrophages and in Apoe(-/-) mice bone-marrow-derived macrophages with low and high basal levels of TGs. Perilipin-3 was only synthesized in mice macrophages with low basal levels of TGs. The regulation was dependent on the fatty acid composition of the lipoproteins; monounsaturated and polyunsaturated fatty acids (PUFAs) more strongly attenuated these effects compared with saturated fatty acids. In THP-1 macrophages, immunofluorescence microscopy and freeze-fracture immunogold labeling indicated that the lipoproteins translocated perilipin-3 from the cytoplasm to the LD surface; only the lipoproteins that were rich in PUFAs suppressed this effect. Chemical inhibition showed that lipoproteins induced perilipin-2 protein expression through the peroxisome proliferator-activated nuclear receptor (PPAR) PPARα and PPARγ pathways. Overall, our data indicate that postprandial TRLs may be involved in atherosclerotic plaque formation through the regulation of perilipin-2 and perilipin-3 proteins in macrophages. Because the fatty acid composition of the lipoproteins is dependent on the type of fat consumed, the ingestion of olive oil, which is rich in monounsaturated fatty acids, and fish oil, which is rich in omega-3 fatty acids, can be considered a good nutritional strategy to reduce the risk of atherosclerosis by LD-associated proteins decrease. Copyright © 2015 Elsevier Inc. All rights reserved.

  13. Moessbauer spectroscopic studies of iron-storage proteins

    Energy Technology Data Exchange (ETDEWEB)

    St. Pierre, T.G.

    1986-01-01

    /sup 57/Fe Moessbauer spectroscopy was used to study iron storage proteins. Various cryostats and a superconducting magnet were used to obtain sample environment temperatures from 1.3 to 200K and applied magnetic fields of up to 10T. The Moessbauer spectra of ferritins isolated from iron-overloaded human spleen, limpet (Patella vulgata), giant limpet (Patella laticostata) and chiton (Clavarizona hirtosa) hemolymph, and bacterial (Pseudomonas aeruginosa) cells are used to gain information on the magnetic ordering- and superparamagnetic transition temperatures of the microcrystalline cores of the proteins. Investigations were made about the cause of the difference in the magnetic anisotropy constants of the cores of iron-overloaded human spleen ferritin and hemosiderin. Livers taken from an iron-overloaded hornbill and artificially iron-loaded rats showed no component with a superparamagnetic transition temperature approaching that of the human spleen hemosiderin.

  14. Perilipin-mediated lipid droplet formation in adipocytes promotes sterol regulatory element-binding protein-1 processing and triacylglyceride accumulation.

    Directory of Open Access Journals (Sweden)

    Yu Takahashi

    Full Text Available Sterol regulatory element-binding protein-1 (SREBP-1 has been thought to be a critical factor that assists adipogenesis. During adipogenesis SREBP-1 stimulates lipogenic gene expression, and peroxisome proliferator-activated receptor γ (PPARγ enhances perilipin (plin gene expression, resulting in generating lipid droplets (LDs to store triacylglycerol (TAG in adipocytes. Plin coats adipocyte LDs and protects them from lipolysis. Here we show in white adipose tissue (WAT of plin-/- mice that nuclear active SREBP-1 and its target gene expression, but not nuclear SREBP-2, significantly decreased on attenuated LD formation. When plin-/- mouse embryonic fibroblasts (MEFs differentiated into adipocytes, attenuated LDs were formed and nuclear SREBP-1 decreased, but enforced plin expression restored them to their original state. Since LDs are largely derived from the endoplasmic reticulum (ER, alterations in the ER cholesterol content were investigated during adipogenesis of 3T3-L1 cells. The ER cholesterol greatly reduced in differentiated adipocytes. The ER cholesterol level in plin-/- WAT was significantly higher than that of wild-type mice, suggesting that increased LD formation caused a change in ER environment along with a decrease in cholesterol. When GFP-SREBP-1 fusion proteins were exogenously expressed in 3T3-L1 cells, a mutant protein lacking the S1P cleavage site was poorly processed during adipogenesis, providing evidence of the increased canonical pathway for SREBP processing in which SREBP-1 is activated by two cleavage enzymes in the Golgi. Therefore, LD biogenesis may create the ER microenvironment favorable for SREBP-1 activation. We describe the novel interplay between LD formation and SREBP-1 activation through a positive feedback loop.

  15. Droplet based microfluidics

    International Nuclear Information System (INIS)

    Seemann, Ralf; Brinkmann, Martin; Pfohl, Thomas; Herminghaus, Stephan

    2012-01-01

    Droplet based microfluidics is a rapidly growing interdisciplinary field of research combining soft matter physics, biochemistry and microsystems engineering. Its applications range from fast analytical systems or the synthesis of advanced materials to protein crystallization and biological assays for living cells. Precise control of droplet volumes and reliable manipulation of individual droplets such as coalescence, mixing of their contents, and sorting in combination with fast analysis tools allow us to perform chemical reactions inside the droplets under defined conditions. In this paper, we will review available drop generation and manipulation techniques. The main focus of this review is not to be comprehensive and explain all techniques in great detail but to identify and shed light on similarities and underlying physical principles. Since geometry and wetting properties of the microfluidic channels are crucial factors for droplet generation, we also briefly describe typical device fabrication methods in droplet based microfluidics. Examples of applications and reaction schemes which rely on the discussed manipulation techniques are also presented, such as the fabrication of special materials and biophysical experiments.

  16. Human Adipose-Derived Mesenchymal Stem/Stromal Cells Handling Protocols. Lipid Droplets and Proteins Double-Staining

    Directory of Open Access Journals (Sweden)

    Aldana D. Gojanovich

    2018-04-01

    Full Text Available Human Adipose-derived mesenchymal stem/stromal cells (hASCs are of great interest because of their potential for therapeutic approaches. The method described here covers every single step necessary for hASCs isolation from subcutaneous abdominal adipose tissue, multicolor phenotyping by flow cytometry, and quantitative determination of adipogenic differentiation status by means of lipid droplets (LDs accumulation, and Western blot analysis. Moreover, to simultaneously analyze both LDs accumulation and cellular proteins localization by fluorescence microscopy, we combined Oil Red O (ORO staining with immunofluorescence detection. For LDs quantification we wrote a program for automatic ORO-stained digital image processing implemented in Octave, a freely available software package. Our method is based on the use of the traditional low cost neutral lipids dye ORO, which can be imaged both by bright-field and fluorescence microscopy. The utilization of ORO instead of other more expensive lipid-specific dyes, together with the fact that the whole method has been designed employing cost-effective culture reagents (standard culture medium and serum, makes it affordable for tight-budget research laboratories. These may be replaced, if necessary or desired, by defined xeno-free reagents for clinical research and applications.

  17. Lipid droplet-binding protein TIP47 regulates hepatitis C Virus RNA replication through interaction with the viral NS5A protein.

    Directory of Open Access Journals (Sweden)

    Dorothee A Vogt

    Full Text Available The nonstructural protein NS5A has emerged as a new drug target in antiviral therapies for Hepatitis C Virus (HCV infection. NS5A is critically involved in viral RNA replication that takes place at newly formed membranes within the endoplasmic reticulum (membranous web and assists viral assembly in the close vicinity of lipid droplets (LDs. To identify host proteins that interact with NS5A, we performed a yeast two-hybrid screen with the N-terminus of NS5A (amino acids 1-31, a well-studied α-helical domain important for the membrane tethering of NS5A. Our studies identified the LD-associated host protein, Tail-Interacting Protein 47 (TIP47 as a novel NS5A interaction partner. Coimmunoprecipitation experiments in Huh7 hepatoma cells confirmed the interaction of TIP47 with full-length NS5A. shRNA-mediated knockdown of TIP47 caused a more than 10-fold decrease in the propagation of full-length infectious HCV in Huh7.5 hepatoma cells. A similar reduction was observed when TIP47 was knocked down in cells harboring an autonomously replicating HCV RNA (subgenomic replicon, indicating that TIP47 is required for efficient HCV RNA replication. A single point mutation (W9A in NS5A that disrupts the interaction with TIP47 but preserves proper subcellular localization severely decreased HCV RNA replication. In biochemical membrane flotation assays, TIP47 cofractionated with HCV NS3, NS5A, NS5B proteins, and viral RNA, and together with nonstructural viral proteins was uniquely distributed to lower-density LD-rich membrane fractions in cells actively replicating HCV RNA. Collectively, our data support a model where TIP47--via its interaction with NS5A--serves as a novel cofactor for HCV infection possibly by integrating LD membranes into the membranous web.

  18. Inhibition of HCV replication by oxysterol-binding protein-related protein 4 (ORP4 through interaction with HCV NS5B and alteration of lipid droplet formation.

    Directory of Open Access Journals (Sweden)

    In-Woo Park

    Full Text Available Hepatitis C virus (HCV RNA replication involves complex interactions among the 3'x RNA element within the HCV 3' untranslated region, viral and host proteins. However, many of the host proteins remain unknown. In this study, we devised an RNA affinity chromatography /2D/MASS proteomics strategy and identified nine putative 3' X-associated host proteins; among them is oxysterol-binding protein-related protein 4 (ORP4, a cytoplasmic receptor for oxysterols. We determined the relationship between ORP4 expression and HCV replication. A very low level of constitutive ORP4 expression was detected in hepatocytes. Ectopically expressed ORP4 was detected in the endoplasmic reticulum and inhibited luciferase reporter gene expression in HCV subgenomic replicon cells and HCV core expression in JFH-1-infected cells. Expression of ORP4S, an ORP4 variant that lacked the N-terminal pleckstrin-homology domain but contained the C-terminal oxysterol-binding domain also inhibited HCV replication, pointing to an important role of the oxysterol-binding domain in ORP4-mediated inhibition of HCV replication. ORP4 was found to associate with HCV NS5B and its expression led to inhibition of the NS5B activity. ORP4 expression had little effect on intracellular lipid synthesis and secretion, but it induced lipid droplet formation in the context of HCV replication. Taken together, these results demonstrate that ORP4 is a negative regulator of HCV replication, likely via interaction with HCV NS5B in the replication complex and regulation of intracellular lipid homeostasis. This work supports the important role of lipids and their metabolism in HCV replication and pathogenesis.

  19. Droplet Growth

    Science.gov (United States)

    Marder, Michael Paolo

    When a mixture of two materials, such as aluminum and tin, or alcohol and water, is cooled below a certain temperature, the two components begin to separate. If one component is dilute in the other, it may separate out in the form of small spheres, and these will begin to enlarge, depleting the supersaturated material around them. If the dynamics is sufficiently slow, thermodynamics gives one considerable information about how the droplets grow. Two types of experiment have explored this behavior and given puzzling results. Nucleation experiments measure the rate at which droplets initially appear from a seemingly homogeneous mixture. Near the critical point in binary liquids, experiments conducted in the 1960's and early 1970's showed that nucleation was vastly slower than theory seemed to predict. The resolution of this problem arises by considering in detail the dynamics of growing droplets and comparing it with what experiments actually measure. Here will be presented a more detailed comparison of theory and experiment than has before been completed, obtaining satisfactory agreement with no free parameters needed. A second type of experiment measures droplet size distributions after long times. In the late stage, droplets compete with each other for material, a few growing at the expense of others. A theory first proposed by Lifshitz and Slyozov claims that this distribution, properly scaled, should be universal, and independent of properties of materials. Yet experimental measurements consistently find distributions that are more broad and squat than the theory would predict. Satisfactory agreement with experiment can be achieved by considering two points. First, one must study the complete time development of droplet size distributions, to understand when the asymptotic regime obtains. Second, droplet size distributions are spread by correlations between droplets. If one finds a small droplet, it is small because large droplets nearby are competing with it

  20. Effects of Long-Term Storage Time and Original Sampling Month on Biobank Plasma Protein Concentrations

    Directory of Open Access Journals (Sweden)

    Stefan Enroth

    2016-10-01

    Full Text Available The quality of clinical biobank samples is crucial to their value for life sciences research. A number of factors related to the collection and storage of samples may affect the biomolecular composition. We have studied the effect of long-time freezer storage, chronological age at sampling, season and month of the year and on the abundance levels of 108 proteins in 380 plasma samples collected from 106 Swedish women. Storage time affected 18 proteins and explained 4.8–34.9% of the observed variance. Chronological age at sample collection after adjustment for storage-time affected 70 proteins and explained 1.1–33.5% of the variance. Seasonal variation had an effect on 15 proteins and month (number of sun hours affected 36 proteins and explained up to 4.5% of the variance after adjustment for storage-time and age. The results show that freezer storage time and collection date (month and season exerted similar effect sizes as age on the protein abundance levels. This implies that information on the sample handling history, in particular storage time, should be regarded as equally prominent covariates as age or gender and need to be included in epidemiological studies involving protein levels.

  1. Changes in cod muscle proteins during frozen storage revealed by proteome analysis and multivariate data analysis

    DEFF Research Database (Denmark)

    Kjærsgård, Inger Vibeke Holst; Nørrelykke, M.R.; Jessen, Flemming

    2006-01-01

    Multivariate data analysis has been combined with proteomics to enhance the recovery of information from 2-DE of cod muscle proteins during different storage conditions. Proteins were extracted according to 11 different storage conditions and samples were resolved by 2-DE. Data generated by 2-DE...... was subjected to principal component analysis (PCA) and discriminant partial least squares regression (DPLSR). Applying PCA to 2-DE data revealed the samples to form groups according to frozen storage time, whereas differences due to different storage temperatures or chilled storage in modified atmosphere...... light chain 1, 2 and 3, triose-phosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase, aldolase A and two ?-actin fragments, and a nuclease diphosphate kinase B fragment to change in concentration, during frozen storage. Application of proteomics, multivariate data analysis and MS/MS to analyse...

  2. Seed storage protein polymorphism in ten elite rice (Oryza sativa L ...

    African Journals Online (AJOL)

    user

    2011-02-14

    Feb 14, 2011 ... for several economical traits by conserving landrace genotypes and ... plasm, seed storage protein analysis represents a valid alternative ... of each variety was taken and ground into fine powder using pestle and mortal and ...

  3. Characterization of Seed Storage Proteins from Chickpea Using 2D Electrophoresis Coupled with Mass Spectrometry

    Directory of Open Access Journals (Sweden)

    Pramod Kumar Singh

    2016-01-01

    Full Text Available Proteomic analysis was employed to map the seed storage protein network in landrace and cultivated chickpea accessions. Protein extracts were separated by two-dimensional gel electrophoresis (2D-GE across a broad range 3.0–10.0 immobilized pH gradient (IPG strips. Comparative elucidation of differentially expressed proteins between two diverse geographically originated chickpea accessions was carried out using 2D-GE coupled with mass spectrometry. A total of 600 protein spots were detected in these accessions. In-gel protein expression patterns revealed three protein spots as upregulated and three other as downregulated. Using trypsin in-gel digestion, these differentially expressed proteins were identified by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS which showed 45% amino acid homology of chickpea seed storage proteins with Arabidopsis thaliana.

  4. Characterization of Seed Storage Proteins from Chickpea Using 2D Electrophoresis Coupled with Mass Spectrometry

    OpenAIRE

    Singh, Pramod Kumar; Shrivastava, Nidhi; Chaturvedi, Krishna; Sharma, Bechan; Bhagyawant, Sameer S.

    2016-01-01

    Proteomic analysis was employed to map the seed storage protein network in landrace and cultivated chickpea accessions. Protein extracts were separated by two-dimensional gel electrophoresis (2D-GE) across a broad range 3.0–10.0 immobilized pH gradient (IPG) strips. Comparative elucidation of differentially expressed proteins between two diverse geographically originated chickpea accessions was carried out using 2D-GE coupled with mass spectrometry. A total of 600 protein spots were detected ...

  5. Mechanism of petroleum-induced sex-specific protein droplet nephropathy and renal cell proliferation in Fischer-344 rats: relevance to humans

    International Nuclear Information System (INIS)

    Charbonneau, M.; Short, B.G.; Lock, E.A.; Swenberg, J.A.

    1987-01-01

    Acute inhalation exposure of male rats to vaporized unleaded gasoline causes a protein droplet-nephropathy syndrome, whereas chronic exposure produces a significant increase renal tumor incidence. The renal lesions produced by chronic or acute exposure to UG have not been observed in kidneys of female rats, or either sex of mice. The assessment of the genotoxic properties of unleaded gasoline by a battery of tests has shown that unleaded gasoline is non-genotoxic. A 21-day histoautoradiographic study in male rats exposed to inhaled unleaded gasoline or gavaged with 2,2,4-trimethylpentane (TMP), a nephrotoxic component of unleaded gasoline selected as a model compound, has shown a dose-dependent increase in cell proliferation specifically in the proximal tubule, segments that have an increased protein droplet formation. A disposition study in male and female rats showed that after a single dose of [ 14 C]-TMP, TMP-derived radiolabel was retained in kidneys of male rats. An increase in the renal α2u-globulin concentration was concomitantly observed in male but not female rats

  6. The collaborative work of droplet assembly.

    Science.gov (United States)

    Chen, Xiao; Goodman, Joel M

    2017-10-01

    Three proteins have been implicated in the assembly of cytoplasmic lipid droplets: seipin, FIT2, and perilipin. This review examines the current theories of seipin function as well as the evidence for the involvement of all three proteins in droplet biogenesis, and ends with a proposal of how they collaborate to regulate the formation of droplets. This article is part of a Special Issue entitled: Recent Advances in Lipid Droplet Biology edited by Rosalind Coleman and Matthijs Hesselink. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Postnatal Deletion of Fat Storage-inducing Transmembrane Protein 2 (FIT2/FITM2) Causes Lethal Enteropathy.

    Science.gov (United States)

    Goh, Vera J; Tan, Jolene S Y; Tan, Bryan C; Seow, Colin; Ong, Wei-Yi; Lim, Yen Ching; Sun, Lei; Ghosh, Sujoy; Silver, David L

    2015-10-16

    Lipid droplets (LDs) are phylogenetically conserved cytoplasmic organelles that store neutral lipids within a phospholipid monolayer. LDs compartmentalize lipids and may help to prevent cellular damage caused by their excess or bioactive forms. FIT2 is a ubiquitously expressed transmembrane endoplasmic reticulum (ER) membrane protein that has previously been implicated in LD formation in mammalian cells and tissue. Recent data indicate that FIT2 plays an essential role in fat storage in an in vivo constitutive adipose FIT2 knock-out mouse model, but the physiological effects of postnatal whole body FIT2 depletion have never been studied. Here, we show that tamoxifen-induced FIT2 deletion using a whole body ROSA26CreER(T2)-driven FIT2 knock-out (iF2KO) mouse model leads to lethal intestinal pathology, including villus blunting and death of intestinal crypts, and loss of lipid absorption. iF2KO mice lose weight and die within 2 weeks after the first tamoxifen dose. At the cellular level, LDs failed to form in iF2KO enterocytes after acute oil challenge and instead accumulated within the ER. Intestinal bile acid transporters were transcriptionally dysregulated in iF2KO mice, leading to the buildup of bile acids within enterocytes. These data support the conclusion that FIT2 plays an essential role in regulating intestinal health and survival postnatally. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  8. Membrane fusion between baculovirus budded virus-enveloped particles and giant liposomes generated using a droplet-transfer method for the incorporation of recombinant membrane proteins.

    Science.gov (United States)

    Nishigami, Misako; Mori, Takaaki; Tomita, Masahiro; Takiguchi, Kingo; Tsumoto, Kanta

    2017-07-01

    Giant proteoliposomes are generally useful as artificial cell membranes in biochemical and biophysical studies, and various procedures for their preparation have been reported. We present here a novel preparation technique that involves the combination of i) cell-sized lipid vesicles (giant unilamellar vesicles, GUVs) that are generated using the droplet-transfer method, where lipid monolayer-coated water-in-oil microemulsion droplets interact with oil/water interfaces to form enclosed bilayer vesicles, and ii) budded viruses (BVs) of baculovirus (Autographa californica nucleopolyhedrovirus) that express recombinant transmembrane proteins on their envelopes. GP64, a fusogenic glycoprotein on viral envelopes, is activated by weak acids and is thought to cause membrane fusion with liposomes. Using confocal laser scanning microscopy (CLSM), we observed that the single giant liposomes fused with octadecyl rhodamine B chloride (R18)-labeled wild-type BV envelopes with moderate leakage of entrapped soluble compounds (calcein), and the fusion profile depended on the pH of the exterior solution: membrane fusion occurred at pH ∼4-5. We further demonstrated that recombinant transmembrane proteins, a red fluorescent protein (RFP)-tagged GPCR (corticotropin-releasing hormone receptor 1, CRHR1) and envelope protein GP64 could be partly incorporated into membranes of the individual giant liposomes with a reduction of the pH value, though there were also some immobile fluorescent spots observed on their circumferences. This combination may be useful for preparing giant proteoliposomes containing the desired membranes and inner phases. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Dynamic changes in proteins during apple (Malus x domestica) fruit ripening and storage

    OpenAIRE

    Shi, Yun; Jiang, Li; Zhang, Li; Kang, Ruoyi; Yu, Zhifang

    2014-01-01

    A proteomic study, using two-dimensional polyacrylamide gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight/time-of-flight, was conducted in apple fruit (cv. ‘Golden Delicious’) starting at 10 days prior to harvest through 50 days in storage. Total protein was extracted using a phenol/sodium dodecyl sulfate protocol. More than 400 protein spots were detected in each gel and 55 differentially expressed proteins (p

  10. Conservation of polypyrimidine tract binding proteins and their putative target RNAs in several storage root crops.

    Science.gov (United States)

    Kondhare, Kirtikumar R; Kumar, Amit; Hannapel, David J; Banerjee, Anjan K

    2018-02-07

    Polypyrimidine-tract binding proteins (PTBs) are ubiquitous RNA-binding proteins in plants and animals that play diverse role in RNA metabolic processes. PTB proteins bind to target RNAs through motifs rich in cytosine/uracil residues to fine-tune transcript metabolism. Among tuber and root crops, potato has been widely studied to understand the mobile signals that activate tuber development. Potato PTBs, designated as StPTB1 and StPTB6, function in a long-distance transport system by binding to specific mRNAs (StBEL5 and POTH1) to stabilize them and facilitate their movement from leaf to stolon, the site of tuber induction, where they activate tuber and root growth. Storage tubers and root crops are important sustenance food crops grown throughout the world. Despite the availability of genome sequence for sweet potato, cassava, carrot and sugar beet, the molecular mechanism of root-derived storage organ development remains completely unexplored. Considering the pivotal role of PTBs and their target RNAs in potato storage organ development, we propose that a similar mechanism may be prevalent in storage root crops as well. Through a bioinformatics survey utilizing available genome databases, we identify the orthologues of potato PTB proteins and two phloem-mobile RNAs, StBEL5 and POTH1, in five storage root crops - sweet potato, cassava, carrot, radish and sugar beet. Like potato, PTB1/6 type proteins from these storage root crops contain four conserved RNA Recognition Motifs (characteristic of RNA-binding PTBs) in their protein sequences. Further, 3´ UTR (untranslated region) analysis of BEL5 and POTH1 orthologues revealed the presence of several cytosine/uracil motifs, similar to those present in potato StBEL5 and POTH1 RNAs. Using RT-qPCR assays, we verified the presence of these related transcripts in leaf and root tissues of these five storage root crops. Similar to potato, BEL5-, PTB1/6- and POTH1-like orthologue RNAs from the aforementioned storage root

  11. Genotypic variability and mutant identification in cicer arietinum L. by seed storage protein profiling

    International Nuclear Information System (INIS)

    Hameed, A.; Iqbal, N.; Shah, T.M.

    2012-01-01

    A collection of thirty-four chickpea genotypes, including five kabuli and twenty-nine desi, were analyzed by SDS-PAGE for seed storage protein profiling. Total soluble seed proteins were resolved on 12% gels. A low level of variability was observed in desi as compared to kabuli genotypes. Dendrogram based on electrophoretic data clustered the thirty-four genotypes in four major groups. As large number of desi genotypes illustrated identical profiles, therefore could not be differentiated on the basis of seed storage protein profiles. One kabuli genotype ILC-195 found to be the most divergent showing 86% similarity with all other genotypes. ILC-195 can be distinguished from its mutant i.e., CM-2000 and other kabuli genotypes on the basis of three peptides i.e. SSP-66, SSP-43 and SSP-39. Some proteins peptides were found to be genotype specific like SSP-26 for ICCV-92311. Uniprot and NCBI protein databases were searched for already reported and characterized seed storage proteins in chickpea. Among 33 observed peptides, only six seed storages proteins from chickpea source were available in databases. On the basis of molecular weight similarity, identified peptides were SSP-64 as Serine/Threonine dehydratase, SSP-56 as Alpha-amylase inhibitor, SSP-50 as Provicillin, SSP-39 as seed imbibition protein, SSP-35 as Isoflavane reductase and SSP-19 as lipid transport protein. Highest variability was observed in vicillin subunits and beta subunits of legumins and its polymorphic forms. In conclusion, seed storage profiling can be economically used to asses the genetic variation, phylogenetic relationship and as markers to differentiate mutants from their parents. (author)

  12. Functions of the CCCH type zinc finger protein OsGZF1 in regulation of the seed storage protein GluB-1 from rice

    NARCIS (Netherlands)

    Chen, Y.; Sun, A.; Wang, M.; Zhu, Z.; Ouwerkerk, P.B.F.

    2014-01-01

    Glutelins are the most abundant storage proteins in rice grain and can make up to 80 % of total protein content. The promoter region of GluB-1, one of the glutelin genes in rice, has been intensively used as a model to understand regulation of seed-storage protein accumulation. In this study, we

  13. The dynamics of milk droplet-droplet collisions

    Science.gov (United States)

    Finotello, Giulia; Kooiman, Roeland F.; Padding, Johan T.; Buist, Kay A.; Jongsma, Alfred; Innings, Fredrik; Kuipers, J. A. M.

    2018-01-01

    Spray drying is an important industrial process to produce powdered milk, in which concentrated milk is atomized into small droplets and dried with hot gas. The characteristics of the produced milk powder are largely affected by agglomeration, combination of dry and partially dry particles, which in turn depends on the outcome of a collision between droplets. The high total solids (TS) content and the presence of milk proteins cause a relatively high viscosity of the fed milk concentrates, which is expected to largely influence the collision outcomes of drops inside the spray. It is therefore of paramount importance to predict and control the outcomes of binary droplet collisions. Only a few studies report on droplet collisions of high viscous liquids and no work is available on droplet collisions of milk concentrates. The current study therefore aims to obtain insight into the effect of viscosity on the outcome of binary collisions between droplets of milk concentrates. To cover a wide range of viscosity values, three milk concentrates (20, 30 and 46% TS content) are investigated. An experimental set-up is used to generate two colliding droplet streams with consistent droplet size and spacing. A high-speed camera is used to record the trajectories of the droplets. The recordings are processed by Droplet Image Analysis in MATLAB to determine the relative velocities and the impact geometries for each individual collision. The collision outcomes are presented in a regime map dependent on the dimensionless impact parameter and Weber ( We) number. The Ohnesorge ( Oh) number is introduced to describe the effect of viscosity from one liquid to another and is maintained constant for each regime map by using a constant droplet diameter ( d ˜ 700 μ m). In this work, a phenomenological model is proposed to describe the boundaries demarcating the coalescence-separation regimes. The collision dynamics and outcome of milk concentrates are compared with aqueous glycerol

  14. Identification and characterisation of seed storage protein transcripts from Lupinus angustifolius

    Directory of Open Access Journals (Sweden)

    Goggin Danica E

    2011-04-01

    Full Text Available Abstract Background In legumes, seed storage proteins are important for the developing seedling and are an important source of protein for humans and animals. Lupinus angustifolius (L., also known as narrow-leaf lupin (NLL is a grain legume crop that is gaining recognition as a potential human health food as the grain is high in protein and dietary fibre, gluten-free and low in fat and starch. Results Genes encoding the seed storage proteins of NLL were characterised by sequencing cDNA clones derived from developing seeds. Four families of seed storage proteins were identified and comprised three unique α, seven β, two γ and four δ conglutins. This study added eleven new expressed storage protein genes for the species. A comparison of the deduced amino acid sequences of NLL conglutins with those available for the storage proteins of Lupinus albus (L., Pisum sativum (L., Medicago truncatula (L., Arachis hypogaea (L. and Glycine max (L. permitted the analysis of a phylogenetic relationships between proteins and demonstrated, in general, that the strongest conservation occurred within species. In the case of 7S globulin (β conglutins and 2S sulphur-rich albumin (δ conglutins, the analysis suggests that gene duplication occurred after legume speciation. This contrasted with 11S globulin (α conglutin and basic 7S (γ conglutin sequences where some of these sequences appear to have diverged prior to speciation. The most abundant NLL conglutin family was β (56%, followed by α (24%, δ (15% and γ (6% and the transcript levels of these genes increased 103 to 106 fold during seed development. We used the 16 NLL conglutin sequences identified here to determine that for individuals specifically allergic to lupin, all seven members of the β conglutin family were potential allergens. Conclusion This study has characterised 16 seed storage protein genes in NLL including 11 newly-identified members. It has helped lay the foundation for efforts to use

  15. The effects of frozen tissue storage conditions on the integrity of RNA and protein.

    Science.gov (United States)

    Auer, H; Mobley, J A; Ayers, L W; Bowen, J; Chuaqui, R F; Johnson, L A; Livolsi, V A; Lubensky, I A; McGarvey, D; Monovich, L C; Moskaluk, C A; Rumpel, C A; Sexton, K C; Washington, M K; Wiles, K R; Grizzle, W E; Ramirez, N C

    2014-10-01

    Unfixed tissue specimens most frequently are stored for long term research uses at either -80° C or in vapor phase liquid nitrogen (VPLN). There is little information concerning the effects such long term storage on tissue RNA or protein available for extraction. Aliquots of 49 specimens were stored for 5-12 years at -80° C or in VPLN. Twelve additional paired specimens were stored for 1 year under identical conditions. RNA was isolated from all tissues and assessed for RNA yield, total RNA integrity and mRNA integrity. Protein stability was analyzed by surface-enhanced or matrix-assisted laser desorption ionization time of flight mass spectrometry (SELDI-TOF-MS, MALDI-TOF-MS) and nano-liquid chromatography electrospray ionization tandem mass spectrometry (nLC-ESI-MS/MS). RNA yield and total RNA integrity showed significantly better results for -80° C storage compared to VPLN storage; the transcripts that were preferentially degraded during VPLN storage were these involved in antigen presentation and processing. No consistent differences were found in the SELDI-TOF-MS, MALDI-TOF-MS or nLC-ESI-MS/MS analyses of specimens stored for more than 8 years at -80° C compared to those stored in VPLN. Long term storage of human research tissues at -80° C provides at least the same quality of RNA and protein as storage in VPLN.

  16. Purification and Initial Functions of Sex-Specific Storage Protein 2 in Bombyx mori.

    Science.gov (United States)

    Chen, Jianqing; Shu, Tejun; Chen, Jian; Ye, Man; Lv, Zhengbing; Nie, Zuoming; Gai, Qijing; Yu, Wei; Zhang, Yaozhou

    2015-08-01

    In this study, we identified a heat-resistant protein from the chrysalis stage of the silkworm which we named sex-specific storage protein 2 (SSP2). This protein was stable even at 80 °C, and has an amino acid sequence that is 90.65 % homologous to SP2. We utilized the heat-resistant characteristics of SSP2 to purify the protein and maintain its biological activity. In addition, using flow cytometry and the MTT assay, we found that SSP2 had anti-apoptotic effects on BmN cells, and that SSP2 could also inhibit cell apoptosis induced by chemical factors. These results suggest that SSP2 has a cell-protective function, and provides a basis for future work on the function of storage proteins in silkworm.

  17. A role for seed storage proteins in Arabidopsis seed longevity

    NARCIS (Netherlands)

    Nguyen, Thu-Phuong|info:eu-repo/dai/nl/328228818; Cueff, Gwendal; Hegedus, Dwayne D; Rajjou, Loïc; Bentsink, Leónie|info:eu-repo/dai/nl/241338735

    2015-01-01

    Proteomics approaches have been a useful tool for determining the biological roles and functions of individual proteins and identifying the molecular mechanisms that govern seed germination, vigour and viability in response to ageing. In this work the dry seed proteome of four Arabidopsis thaliana

  18. Rab32 is important for autophagy and lipid storage in Drosophila.

    Directory of Open Access Journals (Sweden)

    Chao Wang

    Full Text Available Lipids are essential components of all organisms. Within cells, lipids are mainly stored in a specific type of organelle, called the lipid droplet. The molecular mechanisms governing the dynamics of lipid droplets have been little explored. The protein composition of lipid droplets has been analyzed in numerous proteomic studies, and a large number of lipid droplet-associated proteins have been identified, including Rab small GTPases. Rab proteins are known to participate in many intracellular membranous events; however, their exact role in lipid droplets is largely unexplored. Here we systematically investigate the roles of Drosophila Rab family proteins in lipid storage in the larval adipose tissue, fat body. Rab32 and several other Rabs were found to affect the size of lipid droplets as well as lipid levels. Further studies showed that Rab32 and Rab32 GEF/Claret may be involved in autophagy, consequently affecting lipid storage. Loss-of-function mutants of several components in the autophagy pathway result in similar effects on lipid storage. These results highlight the potential functions of Rabs in regulating lipid metabolism.

  19. A phosphatidylinositol transfer protein integrates phosphoinositide signaling with lipid droplet metabolism to regulate a developmental program of nutrient stress-induced membrane biogenesis

    Energy Technology Data Exchange (ETDEWEB)

    Ren, Jihui; Lin, Coney Pei-Chen; Pathak, Manish C.; Temple, Brenda R.S.; Nile, Aaron H.; Mousley, Carl J.; Duncan, Mara C.; Eckert, Debra M.; Leiker, Thomas J.; Ivanova, Pavlina T.; Myers, David S.; Murphy, Robert C.; Brown, H. Alex; Verdaasdonk, Jolien; Bloom, Kerry S.; Ortlund, Eric A.; Neiman, Aaron M.; Bankaitis, Vytas A. [Emory-MED; (SBU); (TAM); (UNC); (Vanderbilt-MED); (Utah); (UCHSC)

    2014-07-11

    Lipid droplet (LD) utilization is an important cellular activity that regulates energy balance and release of lipid second messengers. Because fatty acids exhibit both beneficial and toxic properties, their release from LDs must be controlled. Here we demonstrate that yeast Sfh3, an unusual Sec14-like phosphatidylinositol transfer protein, is an LD-associated protein that inhibits lipid mobilization from these particles. We further document a complex biochemical diversification of LDs during sporulation in which Sfh3 and select other LD proteins redistribute into discrete LD subpopulations. The data show that Sfh3 modulates the efficiency with which a neutral lipid hydrolase-rich LD subclass is consumed during biogenesis of specialized membrane envelopes that package replicated haploid meiotic genomes. These results present novel insights into the interface between phosphoinositide signaling and developmental regulation of LD metabolism and unveil meiosis-specific aspects of Sfh3 (and phosphoinositide) biology that are invisible to contemporary haploid-centric cell biological, proteomic, and functional genomics approaches.

  20. A phosphatidylinositol transfer protein integrates phosphoinositide signaling with lipid droplet metabolism to regulate a developmental program of nutrient stress-induced membrane biogenesis

    Energy Technology Data Exchange (ETDEWEB)

    Ren, Jihui; Lin, Coney Pei-Chen; Pathak, Manish C.; Temple, Brenda R.S.; Nile, Aaron H.; Mousley, Carl J.; Duncan, Mara C.; Eckert, Debra M.; Leiker, Thomas J.; Ivanova, Pavlina T.; Myers, David S.; Murphy, Robert C.; Brown, H. Alex; Verdaasdonk, Jolien; Bloom, Kerry S.; Ortlund, Eric A.; Neiman, Aaron M.; Bankaitis, Vytas A. (Emory-MED); (UNCSM); (UNC); (UCHSC); (TAM); (Vanderbilt-MED); (SBU); (Utah)

    2016-07-06

    Lipid droplet (LD) utilization is an important cellular activity that regulates energy balance and release of lipid second messengers. Because fatty acids exhibit both beneficial and toxic properties, their release from LDs must be controlled. Here we demonstrate that yeast Sfh3, an unusual Sec14-like phosphatidylinositol transfer protein, is an LD-associated protein that inhibits lipid mobilization from these particles. We further document a complex biochemical diversification of LDs during sporulation in which Sfh3 and select other LD proteins redistribute into discrete LD subpopulations. The data show that Sfh3 modulates the efficiency with which a neutral lipid hydrolase-rich LD subclass is consumed during biogenesis of specialized membrane envelopes that package replicated haploid meiotic genomes. These results present novel insights into the interface between phosphoinositide signaling and developmental regulation of LD metabolism and unveil meiosis-specific aspects of Sfh3 (and phosphoinositide) biology that are invisible to contemporary haploid-centric cell biological, proteomic, and functional genomics approaches.

  1. Dengue Virus Uses a Non-Canonical Function of the Host GBF1-Arf-COPI System for Capsid Protein Accumulation on Lipid Droplets.

    Science.gov (United States)

    Iglesias, Nestor G; Mondotte, Juan A; Byk, Laura A; De Maio, Federico A; Samsa, Marcelo M; Alvarez, Cecilia; Gamarnik, Andrea V

    2015-09-01

    Dengue viruses cause the most important human viral disease transmitted by mosquitoes. In recent years, a great deal has been learned about molecular details of dengue virus genome replication; however, little is known about genome encapsidation and the functions of the viral capsid protein. During infection, dengue virus capsid progressively accumulates around lipid droplets (LDs) by an unknown mechanism. Here, we examined the process by which the viral capsid is transported from the endoplasmic reticulum (ER) membrane, where the protein is synthesized, to LDs. Using different methods of intervention, we found that the GBF1-Arf1/Arf4-COPI pathway is necessary for capsid transport to LDs, while the process is independent of both COPII components and Golgi integrity. The transport was sensitive to Brefeldin A, while a drug resistant form of GBF1 was sufficient to restore capsid subcellular distribution in infected cells. The mechanism by which LDs gain or lose proteins is still an open question. Our results support a model in which the virus uses a non-canonical function of the COPI system for capsid accumulation on LDs, providing new ideas for antiviral strategies. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  2. Effect of pomegranate peel extract on lipid and protein oxidation in beef meatballs during refrigerated storage.

    Science.gov (United States)

    Turgut, Sebahattin Serhat; Soyer, Ayla; Işıkçı, Fatma

    2016-06-01

    Antioxidant effect of pomegranate peel extract (PE) to retard lipid and protein oxidation was investigated in meatballs during refrigerated storage at 4±1°C. Concentrated lyophilised water extract of pomegranate peel was incorporated into freshly minced beef meat at 0.5% and 1% concentrations and compared with 0.01% butylated hydroxytoluene (BHT) as a reference and control (without any antioxidant). PE showed high phenolic content and antioxidant activity. In PE added samples, thiobarbituric acid reactive substances (TBARS) value, peroxide formation, loss of sulfhydryl groups and formation of protein carbonyls were lower than control (Pmeatballs prolonged the refrigerated storage up to 8 days. Addition of both 0.5 and 1% PE in meatballs reduced lipid and protein oxidation and improved sensory scores. These results indicated that PE was effective on retarding lipid and protein oxidation. Copyright © 2016 Elsevier Ltd. All rights reserved.

  3. Seed Storage Proteins as a System for Teaching Protein Identification by Mass Spectrometry in Biochemistry Laboratory

    Science.gov (United States)

    Wilson, Karl A.; Tan-Wilson, Anna

    2013-01-01

    Mass spectrometry (MS) has become an important tool in studying biological systems. One application is the identification of proteins and peptides by the matching of peptide and peptide fragment masses to the sequences of proteins in protein sequence databases. Often prior protein separation of complex protein mixtures by 2D-PAGE is needed,…

  4. Storage stability of cauliflower soup powder: The effect of lipid oxidation and protein degradation reactions.

    Science.gov (United States)

    Raitio, Riikka; Orlien, Vibeke; Skibsted, Leif H

    2011-09-15

    Soups based on cauliflower soup powders, prepared by dry mixing of ingredients and rapeseed oil, showed a decrease in quality, as evaluated by a sensory panel, during the storage of the soup powder in the dark for up to 12weeks under mildly accelerated conditions of 40°C and 75% relative humidity. Antioxidant, shown to be effective in protecting the rapeseed bulk oil, used for the powder preparation, had no effect on storage stability of the soup powder. The freshly prepared soup powder had a relatively high concentration of free radicals, as measured by electron spin resonance spectroscopy, which decreased during storage, and most remarkably during the first two weeks of storage, with only marginal increase in lipid hydroperoxides as primary lipid oxidation products, and without any increase in secondary lipid oxidation products. Analyses of volatiles by SPME-GC-MS revealed a significant increase in concentrations of 2-methyl- and 3-methyl butanals, related to Maillard reactions, together with an increase in 2-acetylpyrrole concentration. The soup powders became more brown during storage, as indicated by a decreasing Hunter L-value, in accord with non-enzymatic browning reactions. A significant increase in the concentrations of dimethyl disulfide in soup powder headspace indicated free radical-initiated protein oxidation. Protein degradation, including Maillard reactions and protein oxidation, is concluded to be more important than lipid oxidation in determining the shelf-life of dry cauliflower soup powder. Copyright © 2011 Elsevier Ltd. All rights reserved.

  5. Freezing of Water Droplet due to Evaporation

    Science.gov (United States)

    Satoh, Isao; Fushinobu, Kazuyoshi; Hashimoto, Yu

    In this study, the feasibility of cooling/freezing of phase change.. materials(PCMs) due to evaporation for cold storage systems was experimentally examined. A pure water was used as the test PCM, since the latent heat due to evaporation of water is about 7 times larger than that due to freezing. A water droplet, the diameter of which was 1-4 mm, was suspended in a test cell by a fine metal wire (O. D.= 100μm),and the cell was suddenly evacuated up to the pressure lower than the triple-point pressure of water, so as to enhance the evaporation from the water surface. Temperature of the droplet was measured by a thermocouple, and the cooling/freezing behavior and the temperature profile of the droplet surface were captured by using a video camera and an IR thermo-camera, respectively. The obtained results showed that the water droplet in the evacuated cell is effectively cooled by the evaporation of water itself, and is frozen within a few seconds through remarkable supercooling state. When the initial temperature of the droplet is slightly higher than the room temperature, boiling phenomena occur in the droplet simultaneously with the freezing due to evaporation. Under such conditions, it was shown that the degree of supercooling of the droplet is reduced by the bubbles generated in the droplet.

  6. Nutritional value and digestion rate of rhea meat proteins in association with storage and cooking processes.

    Science.gov (United States)

    Filgueras, Renata S; Gatellier, Philippe; Ferreira, Claude; Zambiazi, Rui C; Santé-Lhoutellier, Véronique

    2011-09-01

    The nutritional value of proteins was investigated after the storage and cooking of rhea M. Gastrocnemius pars interna. Oxidation of basic and aromatic amino acids, surface hydrophobicity and aggregation state of proteins, were determined in raw and cooked meat. In addition, myofibrillar proteins were exposed in vitro to proteases of the digestive tract. Cooking markedly affected the protein surface hydrophobicity. The BBP bound content was three times greater in cooked than in fresh rhea meat. A small increment in tryptophan content after cooking was observed. Storage influenced Schiff bases formation indicating the presence of protein-aldehyde adducts after cooking. High content of Schiff bases was found after cooking of samples stored for 5 days, demonstrating a probable implication of free amino groups, most likely from lysine. Cooking decreased the myofibrillar protein susceptibility to pepsin activity. After cooking, the proteolysis rate by pancreatic enzymes increased. Our findings support the importance of protein aggregation in the nutritional value of meat proteins. Copyright © 2011 Elsevier Ltd. All rights reserved.

  7. Biochemical characterization of amandin, the major storage protein in almond (Prunus dulcis L.).

    Science.gov (United States)

    Sathe, Shridhar K; Wolf, Walter J; Roux, Kenneth H; Teuber, Suzanne S; Venkatachalam, Mahesh; Sze-Tao, Kar Wai Clara

    2002-07-17

    The almond major storage protein, amandin, was prepared by column chromatography (amandin-1), cryoprecipitation (amandin-2), and isoelectric precipitation (amandin-3) methods. Amandin is a legumin type protein characterized by a sedimentation value of 14S. Amandin is composed of two major types of polypeptides with estimated molecular weights of 42-46 and 20-22 kDa linked via disulfide bonds. Several additional minor polypeptides were also present in amandin. Amandin is a storage protein with an estimated molecular weight of 427,300 +/- 47,600 Da (n = 7) and a Stokes radius of 65.88 +/- 3.21 A (n = 7). Amandin is not a glycoprotein. Amandin-1, amandin-2, and amandin-3 are antigenically related and have similar biochemical properties. Amandin-3 is more negatively charged than either amandin-1 or amandin-2. Methionine is the first essential limiting amino acid in amandin followed by lysine and threonine.

  8. Effects of storage conditions on results for quantitative and qualitative evaluation of proteins in canine urine.

    Science.gov (United States)

    Théron, Marie-Laure; Piane, Laetitia; Lucarelli, Laetitia; Henrion, Rémi; Layssol-Lamour, Catherine; Palanché, Florence; Concordet, Didier; Braun, Jean-Pierre D; Trumel, Catherine; Lavoué, Rachel

    2017-08-01

    OBJECTIVE To investigate effects of storage conditions on the canine urine protein-to-creatinine ratio (UPC) and on SDS-agarose gel electrophoresis (AGE) of urinary proteins. SAMPLE Urine specimens from 20 proteinuric (UPC > 0.5) and 20 nonproteinuric (UPC ≤ 0.2) dogs. PROCEDURES UPC and SDS-AGE were performed on urine specimens stored at room temperature (20°C) and 4°C for up to 5 days and at -20° and -80°C for up to 360 days; some specimens were subjected to 3 freeze-thaw cycles. Results were compared with those obtained for fresh urine specimens. RESULTS UPC was not affected by storage at room temperature or by freezing. A decrease in UPC was observed for specimens from nonproteinuric dogs after 5 days at 4°C (10%) and from both groups after 90 days at -20° and -80°C (≤ 20% and ≤ 15%, respectively). The SDS-AGE profiles revealed no visual changes regardless of duration of storage for specimens stored at room temperature, 4°C, and -80°C, except for 1 profile after 360 days at -80°C. Repeated freeze-thaw cycles did not affect SDS-AGE profiles. Appearance or strengthening of high-molecular-weight bands that could alter interpretation was evident in SDS-AGE profiles after storage at -20°C for ≥ 15 days (31/40 dogs). CONCLUSIONS AND CLINICAL RELEVANCE Storage of urine at -20° or -80°C for up to 1 year influenced the UPC without affecting clinical interpretation. Storage of urine specimens at -20°C impaired visual analysis of SDS-AGE. When SDS-AGE cannot be performed on fresh or recently refrigerated urine specimens, storage at -80°C is recommended.

  9. Ammonium accumulation in commercially available embryo culture media and protein supplements during storage at 2-8°C and during incubation at 37°C.

    Science.gov (United States)

    Kleijkers, Sander H M; van Montfoort, Aafke P A; Bekers, Otto; Coonen, Edith; Derhaag, Josien G; Evers, Johannes L H; Dumoulin, John C M

    2016-06-01

    Does ammonium accumulate in commercially available culture media and protein supplements used for in vitro development of human pre-implantation embryos during storage and incubation? Ammonium accumulates in ready-to-use in vitro fertilization (IVF) culture media during storage at 2-8°C and in ready-to-use IVF culture media and protein supplements during incubation at 37°C. Both animal and human studies have shown that the presence of ammonium in culture medium has detrimental effects on embryonic development and pregnancy rate. It is, therefore, important to assess the amount of ammonium accumulation in ready-to-use IVF culture media under conditions that are common in daily practice. Ammonium accumulation was investigated in 15 ready-to-use media, 11 protein-free media and 8 protein supplements. Ammonium was measured by the use of an enzymatic method with glutamate dehydrogenase. To simulate the storage and incubation conditions during IVF treatments, ammonium concentrations were measured at different time-points during storage at 2-8°C for 6 weeks and during incubation at 37°C for 4 days. All ready-to-use, i.e. protein supplemented, culture media showed ammonium accumulation during storage for 6 weeks (ranging from 9.2 to 99.8 µM) and during incubation for 4 days (ranging from 8.4 to 138.6 µM), resulting in levels that might affect embryo development. The protein supplements also showed ammonium accumulation, while the culture media without protein supplementation did not. The main sources of ammonium buildup in ready-to-use culture media were unstable glutamine and the protein supplements. No additional ammonium buildup was found during incubation when using an oil overlay or with the presence of an embryo in the culture droplet. In addition to the unstable glutamine and the protein supplements, other free amino acids might contribute to the ammonium buildup. We did not investigate the deterioration of other components in the media. Break-down of

  10. Irradiation and Post-Irradiation Storage of Chicken: Effects on Fat and Proteins

    International Nuclear Information System (INIS)

    Abou-Tarboush, H.M.; Al-Kahtani, H.A.; Abou-Arab, A.A.; Atia, M.; Bajaber, A.S.; Ahmed, M.A.; El-Mojaddidi, M.A.

    1997-01-01

    Chicken were subjected to gamma irradiation doses of 2.5, 5.0, 7.5 and 10.0 KGy and post-irradiation storage of 21 days at 4±2º. The effects on fat and protein of chicken were studied. Rate of formation of total volatile basic-nitrogen was less in irradiated samples particularly in samples treated with 5.0KGy during the entire storage. Fatty acid profiles of chicken lipids were not significantly (P≤ 0.05) affected by irradiation especially at doses of 5.0 KGy. However, irradiation caused a large increase in thiobarbituric acid (TBA) values which continued gradually during storage. Changes in amino acids were minimal. Irradiated and unirradiated samples showed the appearance of protein subunits with molecular weights in the range of 10.0 to 88.0 and 10.0 to 67.0 KD, respectively. No changes were observed in the sarcoplasmic protein but the intensity of bands in all irradiated samples decreased after 21 days of storage

  11. Effects of gamma irradiation on chickpea seeds vis-a-vis total seed storage proteins, antioxidant activity and protein profiling.

    Science.gov (United States)

    Bhagyawant, S S; Gupta, N; Shrivastava, N

    2015-10-23

    The present work describes radiation—induced effects on seed composition vis—à—vis total seed proteins, antioxidant levels and protein profiling employing two dimensional gel electrophoresis (2D—GE) in kabuli and desi chickpea varities. Seeds were exposed to the radiation doses of 1,2,3,4 and 5 kGy. The total protein concentrations decreased and antioxidant levels were increased with increasing dose compared to control seed samples. Radiation induced effects were dose dependent to these seed parameters while it showed tolerance to 1 kGy dose. Increase in the dose was complimented with increase in antioxidant levels, like 5 kGy enhanced % scavenging activities in all the seed extracts. Precisely, the investigations reflected that the dose range from 2 to 5 kGy was effective for total seed storage proteins, as depicted quantitatively and qualitative 2D—GE means enhance antioxidant activities in vitro.

  12. An inhibition of p38 mitogen activated protein kinase delays the platelet storage lesion.

    Directory of Open Access Journals (Sweden)

    Andrey Skripchenko

    Full Text Available BACKGROUND AND OBJECTIVES: Platelets during storage undergo diverse alterations collectively known as the platelet storage lesion, including metabolic, morphological, functional and structural changes. Some changes correlate with activation of p38 mitogen activated protein kinase (p38 MAPK. Another MAPK, extracellular signal-related kinase (ERK, is involved in PLT activation. The aim of this study was to compare the properties of platelets stored in plasma in the presence or absence of p38 and ERK MAPK inhibitors. MATERIALS AND METHODS: A single Trima apheresis platelet unit (n = 12 was aliquoted into five CLX storage bags. Two aliquots were continuously agitated with or without MAPK inhibitors. Two aliquots were subjected to 48 hours of interruption of agitation with or without MAPK inhibitors. One aliquot contained the same amount of solvent vehicle used to deliver the inhibitor. Platelets were stored at 20-24°C for 7 days and sampled on Days 1, 4, and 7 for 18 in vitro parameters. RESULTS: Inhibition of p38 MAPK by VX-702 leads to better maintenance of all platelet in vitro storage parameters including platelet mitochondrial function. Accelerated by interruption of agitation, the platelet storage lesion of units stored with VX-702 was diminished to that of platelets stored with continuous agitation. Inhibition of ERK MAPK did not ameliorate decrements in any in vitro platelet properties. CONCLUSION: Signaling through p38 MAPK, but not ERK, is associated with platelet deterioration during storage.

  13. Physicochemical properties and storage stability of soybean protein nanoemulsions prepared by ultra-high pressure homogenization.

    Science.gov (United States)

    Xu, Jing; Mukherjee, Dipaloke; Chang, Sam K C

    2018-02-01

    This study investigated the effects of the ultrahigh pressure homogenization (pressure, protein concentration, oil phase fraction, pH, temperature, and ionic strength) and storage on the properties of nanoemulsions (100-500nm range), which were stabilized by laboratory-prepared soybean protein isolate (SPI), β-conglycinin (7S) and glycinin (11S). The nanoemulsions made with SPI, 7S and 11S proteins exhibited considerable stability over various ionic strengths (0-500mM NaCl), pH (7), thermal treatments (30-60°C) and storage (0-45days). The far-UV spectra of SPI, 7S, 11S dispersions, and SPI-, 7S-, 11S protein-stabilized nanoemulsions were analyzed for the protein structural changes following lipid removal. The ultra-high pressure homogenization changed the secondary structure of SPI, 7S, 11S proteins in the nanoemulsions, and enhanced their stability. This study demonstrated that SPI, 7S, and 11S proteins can be used as effective emulsifiers in nanoemulsions prepared by ultra-high pressure homogenization. Copyright © 2017. Published by Elsevier Ltd.

  14. Influence of dietary lipid and protein sources on the sensory quality of organic rainbow trout (Oncorhynchus mykiss) after ice storage

    DEFF Research Database (Denmark)

    Green-Petersen, Ditte; Hyldig, Grethe; Jacobsen, Charlotte

    2014-01-01

    The influence of dietary protein and lipid sources on the quality of organic rainbow trout (Oncorhynchus mykiss) was studied. The protein and oil sources were fishmeal, fish oil, and organic vegetable protein and oils. Sensory profiling was performed during 3 to 14 days of ice storage along...... with lipid analyses of the fillet. Overall, the results showed that the sensory characteristics of the trout were affected in different ways during ice storage. The source of lipid seemed to affect the sensory quality at the beginning of the storage period, while the protein source seemed to have a more...

  15. Effect of pasteurization on the protein composition and oxidative stability of beer during storage.

    Science.gov (United States)

    Lund, Marianne N; Hoff, Signe; Berner, Torben S; Lametsch, René; Andersen, Mogens L

    2012-12-19

    The impacts of pasteurization of a lager beer on protein composition and the oxidative stability were studied during storage at 22 °C for 426 days in the dark. Pasteurization clearly improved the oxidative stability of beer determined by ESR spectroscopy, whereas it had a minor negative effect on the volatile profile by increasing volatile compounds that is generally associated with heat treatment and a loss of fruity ester aroma. A faster rate of radical formation in unpasteurized beer was consistent with a faster consumption of sulfite. Beer proteins in the unpasteurized beer were more degraded, most likely due to proteolytic enzyme activity of yeast remnants and more precipitation of proteins was also observed. The differences in soluble protein content and composition are suggested to result in differences in the contents of prooxidative metals as a consequence of the proteins ability to bind metals. This also contributes to the differences in oxidative stabilities of the beers.

  16. Baking quality parameters of wheat in relation to endosperm storage proteins

    Directory of Open Access Journals (Sweden)

    Daniela Horvat

    2012-01-01

    Full Text Available Wheat storage proteins of twelve winter wheat cultivars grown at the experimental field of the Agricultural Institute Osijek in 2009 were studied for their contribution to the baking quality. Composition of high molecular weight glutenin subunits (HMW-GS was analyzed by SDS-PAGE method, while the proportions of endosperm storage proteins were determined by RP-HPLC method. Regarding the proportion of storage proteins, results of the linear correlation (p<0.05 showed that protein (P and wet gluten (WG content were highly negatively correlated with albumins and globulins (AG and positively with α- gliadins (GLI. A strong negative correlation between AG and water absorption (WA capacity of flour was found, while α- GLI had positive influence on this property. Dough development time (DDT was positively significantly correlated with HMW-GS and negatively with AG. Degree of dough softening (DS was strongly positively affected by γ- GLI and gliadins to glutenins ratio (GLI/GLU and negatively by total GLU and HMW-GS. Dough energy (E and maximum resistance (RMAX were significantly positively affected by Glu-1 score and negatively by GLI/GLU ratio. Resistance to extensibility ratio (R/EXT was significantly negatively correlated with total GLI. Bread volume was significantly negatively influenced by AG.

  17. Influence of yogurt fermentation and refrigerated storage on the stability of protein toxin contaminants.

    Science.gov (United States)

    Jackson, Lauren S; Triplett, Odbert A; Tolleson, William H

    2015-06-01

    Dairy products sold in a ready-to-eat form present the risk that adulterants persisting through manufacturing, storage, and distribution would reach consumers. Pathogenic microbes, including shigatoxigenic strains of Escherichia coli and the toxins they produce, are common food safety hazards associated with dairy products. Ricin and abrin are plant-derived ribosome-inactivating protein toxins related to the shiga-like toxins produced by E. coli. Limited information exists on the effects of manufacturing processes on the stabilities of these heat-resistant ribosome-inactivating proteins in the presence of foods. The goal of this study was to determine how typical yogurt manufacturing and storage processes influence ribosome-inactivating protein toxins. Ricin and abrin were added to skim or whole milk and batch pasteurized. Complete inactivation of both toxins was observed after 30 minutes at 85 °C. If the toxins were added after pasteurization, the levels of ricin and abrin in yogurt and their cytotoxic activities did not change significantly during fermentation or refrigerated storage for 4 weeks. The activities of ricin and abrin were inhibited by skim milk, nonfat yogurt, whole milk, and whole milk yogurt. The results showed minimal effects of the toxins on yogurt pH and %titratable acidity but inhibitory effects of yogurt on toxin activity. Published by Elsevier Ltd.

  18. The Lipid Droplet – A Well-Connected Organelle

    Directory of Open Access Journals (Sweden)

    Qiang eGao

    2015-08-01

    Full Text Available Our knowledge of inter-organellar communication has grown exponentially in recent years. This review focuses on the interactions that cytoplasmic lipid droplets have with other organelles. Twenty-five years ago droplets were considered simply particles of coalesced fat. Ten years ago there were hints from proteomics studies that droplets might interact with other structures to share lipids and proteins. Now it is clear that the droplets interact with many if not most cellular structures to maintain cellular homeostasis and to buffer against insults such as starvation. The evidence for this statement, as well as probes to understand the nature and results of droplet interactions, are presented.

  19. Hypoxia-Inducible Lipid Droplet-Associated Is Not a Direct Physiological Regulator of Lipolysis in Adipose Tissue

    DEFF Research Database (Denmark)

    Dijk, Wieneke; Mattijssen, Frits; de la Rosa Rodriguez, Montserrat

    2017-01-01

    Triglycerides are stored in specialized organelles called lipid droplets. Numerous proteins have been shown to be physically associated with lipid droplets and govern their function. Previously, the protein hypoxia-inducible lipid droplet-associated (HILPDA) was localized to lipid droplets and wa...

  20. Protein mobilities and P-selectin storage in Weibel-Palade bodies.

    Science.gov (United States)

    Kiskin, Nikolai I; Hellen, Nicola; Babich, Victor; Hewlett, Lindsay; Knipe, Laura; Hannah, Matthew J; Carter, Tom

    2010-09-01

    Using fluorescence recovery after photobleaching (FRAP) we measured the mobilities of EGFP-tagged soluble secretory proteins in the endoplasmic reticulum (ER) and in individual Weibel-Palade bodies (WPBs) at early (immature) and late (mature) stages in their biogenesis. Membrane proteins (P-selectin, CD63, Rab27a) were also studied in individual WPBs. In the ER, soluble secretory proteins were mobile; however, following insertion into immature WPBs larger molecules (VWF, Proregion, tPA) and P-selectin became immobilised, whereas small proteins (ssEGFP, eotaxin-3) became less mobile. WPB maturation led to further decreases in mobility of small proteins and CD63. Acute alkalinisation of mature WPBs selectively increased the mobilities of small soluble proteins without affecting larger molecules and the membrane proteins. Disruption of the Proregion-VWF paracrystalline core by prolonged incubation with NH(4)Cl rendered P-selectin mobile while VWF remained immobile. FRAP of P-selectin mutants revealed that immobilisation most probably involves steric entrapment of the P-selectin extracellular domain by the Proregion-VWF paracrystal. Significantly, immobilisation contributed to the enrichment of P-selectin in WPBs; a mutation of P-selectin preventing immobilisation led to a failure of enrichment. Together these data shed new light on the transitions that occur for soluble and membrane proteins following their entry and storage into post-Golgi-regulated secretory organelles.

  1. Protein changes in the albedo of citrus fruits on postharvesting storage.

    Science.gov (United States)

    Lliso, Ignacio; Tadeo, Francisco R; Phinney, Brett S; Wilkerson, Curtis G; Talón, Manuel

    2007-10-31

    In this work, major protein changes in the albedo of the fruit peel of Murcott tangor (tangerine x sweet orange) during postharvest ageing were studied through 2D PAGE. Protein content in matured on-tree fruits and in fruits stored in nonstressing [99% relative humidity (RH) and 25 degrees C], cold (99% RH and 4 degrees C), and drought (60% RH and 25 degrees C) conditions was initially determined. Protein identification through MS/MS determinations revealed in all samples analyzed the occurrence of manganese superoxide dismutase (Mn SOD), actin, ATP synthase beta subunit (ATPase), citrus salt-stress associated protein (CitSap), ascorbate peroxidase (APX), translationally controlled tumor protein (TCTP), and a cysteine proteinase (CP) of the papain family. The latter protein was identified in two different gel spots, with different molecular mass, suggesting the simultaneous presence of the proteinase precursor and its active form. While Mn SOD, actin, ATPase, and CitSap were unchanged in the assayed conditions, TCTP and APX were downregulated during the postharvest ageing process. Ageing-induced APX repression was also reversed by drought. CP contents in albedo, which were similar in on- and off-tree fruits, were strongly dependent upon cold storage. The active/total CP protein ratio significantly increased after cold exposure. This proteomic survey indicates that major changes in protein content in the albedo of the peel of postharvest stored citrus fruits are apparently related to the activation of programmed cell death (PCD).

  2. Effects of storage structures and moisture contents on seed quality attributes of quality protein maize

    Directory of Open Access Journals (Sweden)

    Gopal Bhandari

    2017-12-01

    Full Text Available The study was aimed to examine the effects of various storage structures and moisture contents on seed quality attributes of quality protein maize seed. The quality protein maize (QPM-1 seed was tested in conventional seed storage containers (Fertilizer sack and earthen pot and the improved hermetic ones (Metal bin, Super grain bag, and Purdue Improved Crop Storage (PICS bag at Seed Science and Technology Division, Khumaltar, Nepal during February, 2015 to January 2016. Ten treatments comprising 5 storage devices in two moisture regimes (11% and 9% replicated thrice and laid out in Completely Randomized Design (CRD. Data on temperature, relative humidity (RH, germination, electrical conductivity (EC, seed moisture content (MC were collected bimonthly. The conventional containers were found liable to the external environmental condition whereas the hermetic structures observed with controlled RH level below 40% in all combinations. Electrical conductivity (EC for seed vigor showed that hermetic containers provide higher seed vigor than the conventional ones. Up to 4 months all treatments were found statistically at par for germination. A significant difference was observed in each treatment after 4 months where PICS bag & Super grain bag showed best germination followed by metal bin while fertilizer bag & earthen-pot showed poorer and poorest germination respectively till one year. Almost all treatments with lower MC showed better results than the treatments with higher MC. A negative correlation (R2=69.7% was found between EC and Germination. All six figures from 2 to 12 months on MC showed statistically different where hermetic plastic bags were found maintaining MC as initial whereas MC of fertilizer bags and earthen pot was spiked than the basal figure. The finding evidenced that the hermetic containers and low MC are the seed storage approaches for retaining the quality of seed even in an ambient environmental condition for more than a year.

  3. The FATP1-DGAT2 complex facilitates lipid droplet expansion at the ER-lipid droplet interface

    OpenAIRE

    Farese, Robert; Xu, N; Zhang, SO; Cole, RA; McKinney, SA; Guo, F; Haas, JT; Bobba, S; Farese, RV; Mak, HY

    2012-01-01

    At the subcellular level, fat storage is confined to the evolutionarily conserved compartments termed lipid droplets (LDs), which are closely associated with the endoplasmic reticulum (ER). However, the molecular mechanisms that enable ER-LD interaction an

  4. Convergent evolution of plant and animal embryo defences by hyperstable non-digestible storage proteins.

    Science.gov (United States)

    Pasquevich, María Yanina; Dreon, Marcos Sebastián; Qiu, Jian-Wen; Mu, Huawei; Heras, Horacio

    2017-11-20

    Plants have evolved sophisticated embryo defences by kinetically-stable non-digestible storage proteins that lower the nutritional value of seeds, a strategy that have not been reported in animals. To further understand antinutritive defences in animals, we analysed PmPV1, massively accumulated in the eggs of the gastropod Pomacea maculata, focusing on how its structure and structural stability features affected its capacity to withstand passage through predator guts. The native protein withstands >50 min boiling and resists the denaturing detergent sodium dodecyl sulphate (SDS), indicating an unusually high structural stability (i.e., kinetic stability). PmPV1 is highly resistant to in vitro proteinase digestion and displays structural stability between pH 2.0-12.0 and 25-85 °C. Furthermore, PmPV1 withstands in vitro and mice digestion and is recovered unchanged in faeces, supporting an antinutritive defensive function. Subunit sequence similarities suggest a common origin and tolerance to mutations. This is the first known animal genus that, like plant seeds, lowers the nutritional value of eggs by kinetically-stable non-digestible storage proteins that survive the gut of predators unaffected. The selective pressure of the harsh gastrointestinal environment would have favoured their appearance, extending by convergent evolution the presence of plant-like hyperstable antinutritive proteins to unattended reproductive stages in animals.

  5. Targeted modification of storage protein content resulting in improved amino acid composition of barley grain

    DEFF Research Database (Denmark)

    Sikdar, Md. Shafiqul Islam; Bowra, S; Schmidt, Daiana

    2016-01-01

    family members. Analysis of the AA composition of the transgenic lines showed that the level of essential amino acids increased with a concomitant reduction in proline and glutamine. Both the barley C-hordein and wheat ω-gliadin genes proved successful for RNAi-gene mediated suppression of barley C......C-hordein in barley and ω-gliadins in wheat are members of the prolamins protein families. Prolamins are the major component of cereal storage proteins and composed of non-essential amino acids (AA) such as proline and glutamine therefore have low nutritional value. Using double stranded RNAi...... silencing technology directed towards C-hordein we obtained transgenic barley lines with up to 94.7 % reduction in the levels of C-hordein protein relative to the parental line. The composition of the prolamin fraction of the barley parental line cv. Golden Promise was resolved using SDS...

  6. Relationships between storage protein composition, protein content, growing season and flour quaility of bread wheat

    DEFF Research Database (Denmark)

    Faergestad, E.M.; Flaete, N.E.S.; Magnus, E.M.

    2004-01-01

    ;f alleles appear similar on one-dimensional gels, two-dimensional separation of selected samples may suggest that the f components in these alleles are different proteins. Cross-validated partial least squares regression combined with empirical uncertainty estimates (jack-knifing) of the parameters...

  7. Biomarkers in the diagnosis of lysosomal storage disorders: proteins, lipids, and inhibodies.

    Science.gov (United States)

    Aerts, Johannes M F G; Kallemeijn, Wouter W; Wegdam, Wouter; Joao Ferraz, Maria; van Breemen, Marielle J; Dekker, Nick; Kramer, Gertjan; Poorthuis, Ben J; Groener, Johanna E M; Cox-Brinkman, Josanne; Rombach, Saskia M; Hollak, Carla E M; Linthorst, Gabor E; Witte, Martin D; Gold, Henrik; van der Marel, Gijs A; Overkleeft, Herman S; Boot, Rolf G

    2011-06-01

    A biomarker is an analyte indicating the presence of a biological process linked to the clinical manifestations and outcome of a particular disease. In the case of lysosomal storage disorders (LSDs), primary and secondary accumulating metabolites or proteins specifically secreted by storage cells are good candidates for biomarkers. Clinical applications of biomarkers are found in improved diagnosis, monitoring disease progression, and assessing therapeutic correction. These are illustrated by reviewing the discovery and use of biomarkers for Gaucher disease and Fabry disease. In addition, recently developed chemical tools allowing specific visualization of enzymatically active lysosomal glucocerebrosidase are described. Such probes, coined inhibodies, offer entirely new possibilities for more sophisticated molecular diagnosis, enzyme replacement therapy monitoring, and fundamental research.

  8. The pupylation machinery is involved in iron homeostasis by targeting the iron storage protein ferritin.

    Science.gov (United States)

    Küberl, Andreas; Polen, Tino; Bott, Michael

    2016-04-26

    The balance of sufficient iron supply and avoidance of iron toxicity by iron homeostasis is a prerequisite for cellular metabolism and growth. Here we provide evidence that, in Actinobacteria, pupylation plays a crucial role in this process. Pupylation is a posttranslational modification in which the prokaryotic ubiquitin-like protein Pup is covalently attached to a lysine residue in target proteins, thus resembling ubiquitination in eukaryotes. Pupylated proteins are recognized and unfolded by a dedicated AAA+ ATPase (Mycobacterium proteasomal AAA+ ATPase; ATPase forming ring-shaped complexes). In Mycobacteria, degradation of pupylated proteins by the proteasome serves as a protection mechanism against several stress conditions. Other bacterial genera capable of pupylation such as Corynebacterium lack a proteasome, and the fate of pupylated proteins is unknown. We discovered that Corynebacterium glutamicum mutants lacking components of the pupylation machinery show a strong growth defect under iron limitation, which was caused by the absence of pupylation and unfolding of the iron storage protein ferritin. Genetic and biochemical data support a model in which the pupylation machinery is responsible for iron release from ferritin independent of degradation.

  9. The trafficking pathway of a wheat storage protein in transgenic rice endosperm.

    Science.gov (United States)

    Oszvald, Maria; Tamas, Laszlo; Shewry, Peter R; Tosi, Paola

    2014-04-01

    The trafficking of proteins in the endoplasmic reticulum (ER) of plant cells is a topic of considerable interest since this organelle serves as an entry point for proteins destined for other organelles, as well as for the ER itself. In the current work, transgenic rice was used to study the pattern and pathway of deposition of the wheat high molecular weight (HMW) glutenin sub-unit (GS) 1Dx5 within the rice endosperm using specific antibodies to determine whether it is deposited in the same or different protein bodies from the rice storage proteins, and whether it is located in the same or separate phases within these. The protein distribution and the expression pattern of HMW sub-unit 1Dx5 in transgenic rice endosperm at different stages of development were determined using light and electron microscopy after labelling with antibodies. The use of HMW-GS-specific antibodies showed that sub-unit 1Dx5 was expressed mainly in the sub-aleurone cells of the endosperm and that it was deposited in both types of protein body present in the rice endosperm: derived from the ER and containing prolamins, and derived from the vacuole and containing glutelins. In addition, new types of protein bodies were also formed within the endosperm cells. The results suggest that the HMW 1Dx5 protein could be trafficked by either the ER or vacuolar pathway, possibly depending on the stage of development, and that its accumulation in the rice endosperm could compromise the structural integrity of protein bodies and their segregation into two distinct populations in the mature endosperm.

  10. COMPARATIVE DYNAMICS OF PROTEIN DESTRUCTION IN CANNED FOODS IN SAUCE AT DIFFERENT THERMAL TREATMENT REGIMES AND SUBSEQUENT STORAGE

    OpenAIRE

    V. B. Krylova; T. V. Gustova

    2017-01-01

    In the course of investigations, the structural changes in proteins were established, which were associated with the preliminary treatment of meat ingredients, a pH level of the system and parameters of thermal treatment.The pasteurization regimes allowed retaining a protein nitrogen proportion up to 94% by the end of canned food storage duration. Upon sterilization, the losses in protein nitrogen were two times higher. A negative effect of more acidic sauce on preservation of the protein nitr...

  11. A cytosolic copper storage protein provides a second level of copper tolerance in Streptomyces lividans.

    Science.gov (United States)

    Straw, Megan L; Chaplin, Amanda K; Hough, Michael A; Paps, Jordi; Bavro, Vassiliy N; Wilson, Michael T; Vijgenboom, Erik; Worrall, Jonathan A R

    2018-01-24

    Streptomyces lividans has a distinct dependence on the bioavailability of copper for its morphological development. A cytosolic copper resistance system is operative in S. lividans that serves to preclude deleterious copper levels. This system comprises of several CopZ-like copper chaperones and P 1 -type ATPases, predominantly under the transcriptional control of a metalloregulator from the copper sensitive operon repressor (CsoR) family. In the present study, we discover a new layer of cytosolic copper resistance in S. lividans that involves a protein belonging to the newly discovered family of copper storage proteins, which we have named Ccsp (cytosolic copper storage protein). From an evolutionary perspective, we find Ccsp homologues to be widespread in Bacteria and extend through into Archaea and Eukaryota. Under copper stress Ccsp is upregulated and consists of a homotetramer assembly capable of binding up to 80 cuprous ions (20 per protomer). X-ray crystallography reveals 18 cysteines, 3 histidines and 1 aspartate are involved in cuprous ion coordination. Loading of cuprous ions to Ccsp is a cooperative process with a Hill coefficient of 1.9 and a CopZ-like copper chaperone can transfer copper to Ccsp. A Δccsp mutant strain indicates that Ccsp is not required under initial copper stress in S. lividans, but as the CsoR/CopZ/ATPase efflux system becomes saturated, Ccsp facilitates a second level of copper tolerance.

  12. Lack of Globulin Synthesis during Seed Development Alters Accumulation of Seed Storage Proteins in Rice

    Directory of Open Access Journals (Sweden)

    Hye-Jung Lee

    2015-06-01

    Full Text Available The major seed storage proteins (SSPs in rice seeds have been classified into three types, glutelins, prolamins, and globulin, and the proportion of each SSP varies. It has been shown in rice mutants that when either glutelins or prolamins are defective, the expression of another type of SSP is promoted to counterbalance the deficit. However, we observed reduced abundances of glutelins and prolamins in dry seeds of a globulin-deficient rice mutant (Glb-RNAi, which was generated with RNA interference (RNAi-induced suppression of globulin expression. The expression of the prolamin and glutelin subfamily genes was reduced in the immature seeds of Glb-RNAi lines compared with those in wild type. A proteomic analysis of Glb-RNAi seeds showed that the reductions in glutelin and prolamin were conserved at the protein level. The decreased pattern in glutelin was also significant in the presence of a reductant, suggesting that the polymerization of the glutelin proteins via intramolecular disulfide bonds could be interrupted in Glb-RNAi seeds. We also observed aberrant and loosely packed structures in the storage organelles of Glb-RNAi seeds, which may be attributable to the reductions in SSPs. In this study, we evaluated the role of rice globulin in seed development, showing that a deficiency in globulin could comprehensively reduce the expression of other SSPs.

  13. Prolonging hypothermic storage (4 C) of bovine embryos with fish antifreeze protein.

    Science.gov (United States)

    Ideta, Atsushi; Aoyagi, Yoshito; Tsuchiya, Kanami; Nakamura, Yuuki; Hayama, Kou; Shirasawa, Atsushi; Sakaguchi, Kenichiro; Tominaga, Naomi; Nishimiya, Yoshiyuki; Tsuda, Sakae

    2015-01-01

    Embryos obtained via superovulation are necessary for mammalian artificial reproduction, and viability is a key determinant of success. Nonfreezing storage at 4 C is possible, but currently used storage solutions can maintain embryo viability for only 24-48 h. Here we found that 10 mg/ml antifreeze protein (AFP) dissolved in culture medium 199 with 20% (v/v) fetal bovine serum and 25 mM HEPES could keep bovine embryos alive for 10 days at 4 C. We used a recombinant AFP isolated from the notched-fin eelpout (Zoarces elongatus Kner). Photomicroscopy indicated that the AFP-embryo interaction was enhanced at 37 C. Embryos pre-warmed with the AFP solution at 37 C for 60 min maintained high viability, whereas those that were not pre-warmed could live no longer than 7 days. Thus, short-term storage of bovine embryos was achieved by a combination of AFP-containing medium and controlled pre-warming.

  14. Amino acid and protein changes in tilapia and spanish mackerel after irradiation and storage

    Energy Technology Data Exchange (ETDEWEB)

    Al-Kahtani, Hassan A.; Abu-Tarboush, Hamza M.; Atia, Mohamed; Bajaber, Adnan S.; Ahmed, Mohamed A.; El-Mojaddidi, Mohamed A

    1998-01-01

    Some amino acids in tilapia decreased while some others increased when subjected to doses up to 10.0 kGy. However, 10 kGy contributed to a significant reduction in all amino acids of Spanish mackerel. Variations in amino acid contents continued during post-irradiation storage with no consistent trend of increase or decrease. SDS-PAGE of protein from both fish showed 27 bands of subunits with MW < 14.0-94.0 KD. Isoelectric focusing patterns of sarcoplasmic protein of unirradiated and irradiated fish showed no charge in the number of bands, while some changes were observed in the intensities of the anodic and cathodic bands depending on isoelectric points (pIs)

  15. Effects of water deficit on breadmaking quality and storage protein compositions in bread wheat (Triticum aestivum L.).

    Science.gov (United States)

    Zhou, Jiaxing; Liu, Dongmiao; Deng, Xiong; Zhen, Shoumin; Wang, Zhimin; Yan, Yueming

    2018-03-12

    Water deficiency affects grain proteome dynamics and storage protein compositions, resulting in changes in gluten viscoelasticity. In this study, the effects of field water deficit on wheat breadmaking quality and grain storage proteins were investigated. Water deficiency produced a shorter grain-filling period, a decrease in grain number, grain weight and grain yield, a reduced starch granule size and increased protein content and glutenin macropolymer contents, resulting in superior dough properties and breadmaking quality. Reverse phase ultra-performance liquid chromatography analysis showed that the total gliadin and glutenin content and the accumulation of individual components were significantly increased by water deficiency. Two-dimensional gel electrophoresis detected 144 individual storage protein spots with significant accumulation changes in developing grains under water deficit. Comparative proteomic analysis revealed that water deficiency resulted in significant upregulation of 12 gliadins, 12 high-molecular-weight glutenin subunits and 46 low-molecular-weight glutenin subunits. Quantitative real-time polymerase chain reaction analysis revealed that the expression of storage protein biosynthesis-related transcription factors Dof and Spa was upregulated by water deficiency. The present results illustrated that water deficiency leads to increased accumulation of storage protein components and upregulated expression of Dof and Spa, resulting in an improvement in glutenin strength and breadmaking quality. © 2018 Society of Chemical Industry. © 2018 Society of Chemical Industry.

  16. Integrated transcriptomics and proteomics analysis of storage protein composition in developing barley grain to improve nutritional profile

    DEFF Research Database (Denmark)

    Kaczmarczyk, Agnieszka Ewa; Dionisio, Giuseppe; Renaut, Jenny

    2012-01-01

    The aim of the study was to understand the molecular and biochemical mechanisms underpinning the effect of nitrogen (N) on barley (Hordeum vulgare) storage protein production (hordeins) during grain filling. Using a combination of advanced biochemistry methods, we could comprehensively describe......-regimes caused significant differences in both quantity and quality of the storage proteins transcripts. Principal Component Analysis of the amino acid (AA) profiles also indicated dissimilarity in individual AA percentages, correlated to hordein content. The abundance values of proteins of interest confirmed...

  17. Lipid droplet analysis in caveolin-deficient adipocytes: alterations in surface phospholipid composition and maturation defects

    OpenAIRE

    Blouin, Cedric M.; Le Lay, Soazig; Eberl, Anita; Koefeler, Harald C.; Guerrera, Ida Chiara; Klein, Christophe; Le Liepvre, Xavier; Lasnier, Francoise; Bourron, Olivier; Gautier, Jean-Francois; Ferre, Pascal; Hajduch, Eric; Dugail, Isabelle

    2010-01-01

    Caveolins form plasmalemnal invaginated caveolae. They also locate around intracellular lipid droplets but their role in this location remains unclear. By studying primary adipocytes that highly express caveolin-1, we characterized the impact of caveolin-1 deficiency on lipid droplet proteome and lipidome. We identified several missing proteins on the lipid droplet surface of caveolin-deficient adipocytes and showed that the caveolin-1 lipid droplet pool is organized as multi-protein complexe...

  18. Profiling an electrospray plume by laser-induced fluorescence and Fraunhofer diffraction combined to mass spectrometry: influence of size and composition of droplets on charge-state distributions of electrosprayed proteins.

    Science.gov (United States)

    Girod, Marion; Dagany, Xavier; Boutou, Véronique; Broyer, Michel; Antoine, Rodolphe; Dugourd, Philippe; Mordehai, Alex; Love, Craig; Werlich, Mark; Fjeldsted, John; Stafford, George

    2012-07-14

    We investigated how physico-chemical properties of charged droplets are affected by the electrospray process, using simultaneous in situ measurements by laser-induced fluorescence (LIF), Fraunhofer diffraction and mass spectrometry. For this purpose, we implemented a laser-induced-fluorescence profiling setup in conjunction with a fast, high-resolution particle sizing scheme on a modified Agilent Jet Stream electrospray source coupled to a single quadrupole mass analyser. The optical setup permits us to profile the solvent fractionation and the size of the droplets as they evaporate in an electrospray plume by measuring both the angular scattering pattern and emission spectra of a solvatochromic fluorescent dye. Mass spectra are recorded simultaneously. These mass spectrometry and optical spectroscopy investigations allow us to study the relation between the observed charge-state distributions of protein anions and physico-chemical properties of evaporating droplets in the spray plume. By mixing water with methanol, a refolding of cytochrome C is observed as the water percentage increases in the plume due to the preponderant evaporation of volatile methanol.

  19. Levitated droplet dye laser

    DEFF Research Database (Denmark)

    Azzouz, H.; Alkafadiji, L.; Balslev, Søren

    2006-01-01

    a high quality optical resonator. Our 750 nL lasing droplets consist of Rhodamine 6G dissolved in ethylene glycol, at a concentration of 0.02 M. The droplets are optically pumped at 532 nm light from a pulsed, frequency doubled Nd:YAG laser, and the dye laser emission is analyzed by a fixed grating...

  20. Droplet collisions in turbulence

    NARCIS (Netherlands)

    Oldenziel, G.

    2014-01-01

    Liquid droplets occur in many natural phenomena and play an important role in a large number of industrial applications. One of the distinct properties of droplets as opposed to solid particles is their ability to merge, or coalesce upon collision. Coalescence of liquid drops is of importance in for

  1. REDOX POTENTIAL AND DYNAMICS OF PROTEIN AND FAT DESTRUCTION DURING STORAGE OF CANNED MEAT IN PIECES

    Directory of Open Access Journals (Sweden)

    V. B. Krylova

    2016-01-01

    Full Text Available The studies on the dynamics of the redox potential of systems and its relationship with the processes of protein and fat destruction in canned foods during their storage are fragmented and not systemized, which highlight their topicality. The aim of the research was to obtain the experimental data on the Eh values and physico-chemical indicators of canned food quality during storage in order to establish their possible correlation. It was shown that the dynamics of Eh, the content of free amino acids and fatty acid fractions in the canned products from beef and pork was different during storage. For example, a decrease in the Eh value and free amino acid content in the canned products from beef had a smooth character, while in the canned products from pork several periods were observed, which differed in the character of the change in the quality indicators.A linear character of the changes in the proportion of fatty acid fractions during storage of the canned products from beef and pork was noticed. With that, both canned food items had an increase in the saturated fatty acid content at the concomitant decrease in the sum of mono- and polyunsaturated fatty acids. The value of an increase in the proportion of saturated fatty acids associated with the process of reduction of mono- and polyunsaturated fatty acids did not depend on the kind of meat in the canned foods and was on average 6%. A decrease in the proportion of mono- and polyunsaturated fatty acids in the canned products from pork was about 4 times more intensive compared to the canned products from beef.

  2. Butschli Dynamic Droplet System

    DEFF Research Database (Denmark)

    Armstrong, R.; Hanczyc, M.

    2013-01-01

    Dynamical oil-water systems such as droplets display lifelike properties and may lend themselves to chemical programming to perform useful work, specifically with respect to the built environment. We present Butschli water-in-oil droplets as a model for further investigation into the development...... reconstructed the Butschli system and observed its life span under a light microscope, observing chemical patterns and droplet behaviors in nearly three hundred replicate experiments. Self-organizing patterns were observed, and during this dynamic, embodied phase the droplets provided a means of introducing...... temporal and spatial order in the system with the potential for chemical programmability. The authors propose that the discrete formation of dynamic droplets, characterized by their lifelike behavior patterns, during a variable window of time (from 30 s to 30 min after the addition of alkaline water...

  3. Salvage and storage of infectious disease protein targets in the SSGCID high-throughput crystallization pathway using microfluidics

    International Nuclear Information System (INIS)

    Christensen, Jeff; Gerdts, Cory J.; Clifton, Mathew C.; Stewart, Lance

    2011-01-01

    SSGCID protein crystals were salvaged and stored using the MPCS Plug Maker and CrystalCards when high-throughput traditional sitting-drop vapor diffusion initially failed. The MPCS Plug Maker is a microcapillary-based protein-crystallization system for generating diffraction-ready crystals from nanovolumes of protein. Crystallization screening using the Plug Maker was used as a salvage pathway for proteins that failed to crystallize during the initial observation period using the traditional sitting-drop vapor-diffusion method. Furthermore, the CrystalCards used to store the crystallization experiments set up by the Plug Maker are shown be a viable container for long-term storage of protein crystals without a discernable loss of diffraction quality with time. Use of the Plug Maker with SSGCID proteins is demonstrated to be an effective crystal-salvage and storage method

  4. Fabrication of Flexible, Fully Organic, Degradable Energy Storage Devices Using Silk Proteins.

    Science.gov (United States)

    Pal, Ramendra K; Kundu, Subhas C; Yadavalli, Vamsi K

    2018-03-21

    Flexible and thin-film devices are of great interest in epidermal and implantable bioelectronics. The integration of energy storage and delivery devices such as supercapacitors (SCs) with properties such as flexibility, miniaturization, biocompatibility, and degradability are sought for such systems. Reducing e-waste and using sustainable materials and processes are additional desirable qualities. Herein, a silk protein-based biocompatible and degradable thin-film microSC (μSC) is reported. A protein carrier with the conducting polymer poly(3,4-ethylenedioxythiophene) polystyrene sulfonate and reduced graphene oxide dopant is used as a photopatternable biocomposite ink. Active electrodes are fabricated using photolithography under benign conditions, using only water as the solvent. These electrodes are printed on flexible protein sheets to form degradable, organic devices with a benign agarose-NaCl gel electrolyte. High capacitance, power density, cycling stability over 500 cycles, and the ability to power a light-emitting diode are shown. The device is flexible, can sustain cyclic mechanical stresses over 450 cycles, and retain capacitive properties over several days in liquid. Significantly, the μSCs are cytocompatible and completely degraded over the period of ∼1 month. By precise control of the device configuration, these silk protein-based, all-polymer organic devices can be designed to be tunably transient and provide viable alternatives for powering flexible and implantable bioelectronics.

  5. CHARACTERIZATION OF SEED STORAGE PROTEINS IN SOME IRANIAN DATE PALM CULTIVARS USING SDS-PAGE

    Directory of Open Access Journals (Sweden)

    Sayed Mohammad Reza Khoshroo

    2013-08-01

    Full Text Available The date palm (Phoenix dactylifera L. is most adapted tree to grow in desert areas. It has always been looked on as a key source of stability, survival and evolution of the oasis agro-system since it constitutes the basic features of the ecological pyramid in desert regions. Determining genetic variability and cultivars identification in date palm are two major important factors in breeding programs, characterization of germplasm, and conservation purposes. The genetic variation of seed proteins was assayed by SDS-PAGE for 9 cultivars in Shahdad region in Iran. A total of 16 alternative protein bands with different mobility rates were identified within a molecular weight range of 11 KDa to 350 KDa. Then, electrophorogram for each cultivar was scored, and Jaccard‘s Similarity Index was calculated. Relying on UPGMA and NJ methods, genetic diversity of cultivars was evaluated by constructing the dendrogram for protein bands. Moreover, genetic distance was calculated for all of the cultivars.  It is concluded that seed storage protein profiles could be useful markers in genetic diversity studies and classification of cultivars. The cultivars from Shahdad were well separated from each other. This might have been done due to their unique genetic build-up. The cluster analysis displayed five major classes. In order to precise this assumption, data were computed to perform a PCA. Cluster analysis and PCA demonstrated their validity in establishing genetic diversity. When PCA was studied, the previously described results about Jaccard Similarity Coefficient dendrogram were also visualized.

  6. Chloroform-assisted phenol extraction improving proteome profiling of maize embryos through selective depletion of high-abundance storage proteins.

    Directory of Open Access Journals (Sweden)

    Erhui Xiong

    Full Text Available The presence of abundant storage proteins in plant embryos greatly impedes seed proteomics analysis. Vicilin (or globulin-1 is the most abundant storage protein in maize embryo. There is a need to deplete the vicilins from maize embryo extracts for enhanced proteomics analysis. We here reported a chloroform-assisted phenol extraction (CAPE method for vicilin depletion. By CAPE, maize embryo proteins were first extracted in an aqueous buffer, denatured by chloroform and then subjected to phenol extraction. We found that CAPE can effectively deplete the vicilins from maize embryo extract, allowing the detection of low-abundance proteins that were masked by vicilins in 2-DE gel. The novelty of CAPE is that it selectively depletes abundant storage proteins from embryo extracts of both monocot (maize and dicot (soybean and pea seeds, whereas other embryo proteins were not depleted. CAPE can significantly improve proteome profiling of embryos and extends the application of chloroform and phenol extraction in plant proteomics. In addition, the rationale behind CAPE depletion of abundant storage proteins was explored.

  7. A new look on protein-polyphenol complexation during honey storage: is this a random or organized event with the help of dirigent-like proteins?

    Directory of Open Access Journals (Sweden)

    Katrina Brudzynski

    Full Text Available Honey storage initiates melanoidin formation that involves a cascade of seemingly unguided redox reactions between amino acids/proteins, reducing sugars and polyphenols. In the process, high molecular weight protein-polyphenol complexes are formed, but the mechanism involved remains unknown. The objective of this study was twofold: to determine quantitative and qualitative changes in proteins in honeys stored for prolonged times and in different temperatures and to relate these changes to the formation of protein-polyphenol complexes. Six -month storage decreased the protein content by 46.7% in all tested honeys (t-test, p<0.002 with the rapid reduction occurring during the first three month. The changes in protein levels coincided with alterations in molecular size and net charge of proteins on SDS -PAGE. Electro-blotted proteins reacted with a quinone-specific nitro blue tetrazolium (NBT on nitrocellulose membranes indicating that quinones derived from oxidized polyphenols formed covalent bonds with proteins. Protein-polyphenol complexes isolated by size-exclusion chromatography differed in size and stoichiometry and fall into two categories: (a high molecular weight complexes (230-180 kDa enriched in proteins but possessing a limited reducing activity toward the NBT and (b lower molecular size complexes (110-85 kDa enriched in polyphenols but strongly reducing the dye. The variable stoichiometry suggest that the large, "protein-type" complexes were formed by protein cross-linking, while in the smaller, "polyphenol-type" complexes polyphenols were first polymerized prior to protein binding. Quinones preferentially bound a 31 kDa protein which, by the electrospray quadrupole time of flight mass spectrometry (ESI-Qtof-MS analysis, showed homology to dirigent-like proteins known for assisting in radical coupling and polymerization of phenolic compounds. These findings provide a new look on protein-polyphenol interaction in honey where the

  8. Surfing the wave, cycle, life history, and genes/proteins expressed by testicular germ cells. Part 3: developmental changes in spermatid flagellum and cytoplasmic droplet and interaction of sperm with the zona pellucida and egg plasma membrane.

    Science.gov (United States)

    Hermo, Louis; Pelletier, R-Marc; Cyr, Daniel G; Smith, Charles E

    2010-04-01

    Spermiogenesis constitutes the steps involved in the metamorphosis of spermatids into spermatozoa. It involves modification of several organelles in addition to the formation of several structures including the flagellum and cytoplasmic droplet. The flagellum is composed of a neck region and middle, principal, and end pieces. The axoneme composed of nine outer microtubular doublets circularly arranged to form a cylinder around a central pair of microtubules is present throughout the flagellum. The middle and principal pieces each contain specific components such as the mitochondrial sheath and fibrous sheath, respectively, while outer dense fibers are common to both. A plethora of proteins are constituents of each of these structures, with each playing key roles in functions related to the fertility of spermatozoa. At the end of spermiogenesis, a portion of spermatid cytoplasm remains associated with the released spermatozoa, referred to as the cytoplasmic droplet. The latter has as its main feature Golgi saccules, which appear to modify the plasma membrane of spermatozoa as they move down the epididymal duct and hence may be partly involved in male gamete maturation. The end product of spermatogenesis is highly streamlined and motile spermatozoa having a condensed nucleus equipped with an acrosome. Spermatozoa move through the female reproductive tract and eventually penetrate the zona pellucida and bind to the egg plasma membrane. Many proteins have been implicated in the process of fertilization as well as a plethora of proteins involved in the development of spermatids and sperm, and these are high lighted in this review. Copyright 2009 Wiley-Liss, Inc.

  9. Seed storage protein gene promoters contain conserved DNA motifs in Brassicaceae, Fabaceae and Poaceae

    Science.gov (United States)

    Fauteux, François; Strömvik, Martina V

    2009-01-01

    Background Accurate computational identification of cis-regulatory motifs is difficult, particularly in eukaryotic promoters, which typically contain multiple short and degenerate DNA sequences bound by several interacting factors. Enrichment in combinations of rare motifs in the promoter sequence of functionally or evolutionarily related genes among several species is an indicator of conserved transcriptional regulatory mechanisms. This provides a basis for the computational identification of cis-regulatory motifs. Results We have used a discriminative seeding DNA motif discovery algorithm for an in-depth analysis of 54 seed storage protein (SSP) gene promoters from three plant families, namely Brassicaceae (mustards), Fabaceae (legumes) and Poaceae (grasses) using backgrounds based on complete sets of promoters from a representative species in each family, namely Arabidopsis (Arabidopsis thaliana (L.) Heynh.), soybean (Glycine max (L.) Merr.) and rice (Oryza sativa L.) respectively. We have identified three conserved motifs (two RY-like and one ACGT-like) in Brassicaceae and Fabaceae SSP gene promoters that are similar to experimentally characterized seed-specific cis-regulatory elements. Fabaceae SSP gene promoter sequences are also enriched in a novel, seed-specific E2Fb-like motif. Conserved motifs identified in Poaceae SSP gene promoters include a GCN4-like motif, two prolamin-box-like motifs and an Skn-1-like motif. Evidence of the presence of a variant of the TATA-box is found in the SSP gene promoters from the three plant families. Motifs discovered in SSP gene promoters were used to score whole-genome sets of promoters from Arabidopsis, soybean and rice. The highest-scoring promoters are associated with genes coding for different subunits or precursors of seed storage proteins. Conclusion Seed storage protein gene promoter motifs are conserved in diverse species, and different plant families are characterized by a distinct combination of conserved motifs

  10. Seed storage protein gene promoters contain conserved DNA motifs in Brassicaceae, Fabaceae and Poaceae

    Directory of Open Access Journals (Sweden)

    Fauteux François

    2009-10-01

    Full Text Available Abstract Background Accurate computational identification of cis-regulatory motifs is difficult, particularly in eukaryotic promoters, which typically contain multiple short and degenerate DNA sequences bound by several interacting factors. Enrichment in combinations of rare motifs in the promoter sequence of functionally or evolutionarily related genes among several species is an indicator of conserved transcriptional regulatory mechanisms. This provides a basis for the computational identification of cis-regulatory motifs. Results We have used a discriminative seeding DNA motif discovery algorithm for an in-depth analysis of 54 seed storage protein (SSP gene promoters from three plant families, namely Brassicaceae (mustards, Fabaceae (legumes and Poaceae (grasses using backgrounds based on complete sets of promoters from a representative species in each family, namely Arabidopsis (Arabidopsis thaliana (L. Heynh., soybean (Glycine max (L. Merr. and rice (Oryza sativa L. respectively. We have identified three conserved motifs (two RY-like and one ACGT-like in Brassicaceae and Fabaceae SSP gene promoters that are similar to experimentally characterized seed-specific cis-regulatory elements. Fabaceae SSP gene promoter sequences are also enriched in a novel, seed-specific E2Fb-like motif. Conserved motifs identified in Poaceae SSP gene promoters include a GCN4-like motif, two prolamin-box-like motifs and an Skn-1-like motif. Evidence of the presence of a variant of the TATA-box is found in the SSP gene promoters from the three plant families. Motifs discovered in SSP gene promoters were used to score whole-genome sets of promoters from Arabidopsis, soybean and rice. The highest-scoring promoters are associated with genes coding for different subunits or precursors of seed storage proteins. Conclusion Seed storage protein gene promoter motifs are conserved in diverse species, and different plant families are characterized by a distinct combination

  11. Influence of prolonged storage process, pasteurization, and heat treatment on biologically-active human milk proteins.

    Science.gov (United States)

    Chang, Jih-Chin; Chen, Chao-Huei; Fang, Li-Jung; Tsai, Chi-Ren; Chang, Yu-Chuan; Wang, Teh-Ming

    2013-12-01

    The bioactive proteins in human milk may be influenced by prolonged storage process, pasteurization, and heat treatment. This study was conducted to evaluate the effects of these procedures. Three forms of human milk - freshly expressed, frozen at -20°C for a prolonged duration, and pasteurized milk - were collected from 14 healthy lactating mothers and a milk bank. The concentrations of major bioactive proteins (secretory immunoglobulin A, lactoferrin, lysozyme, and leptin) were quantified using enzyme-linked immunosorbent assay kits. Changes in these proteins by heat treatment at 40°C or 60°C for 30 minutes were further evaluated. The mean concentrations of lactoferrin and secretory immunoglobulin A were significantly reduced by 66% and 25.9%, respectively, in pasteurized milk compared with those in freshly-expressed milk. Heat treatment at 40°C or 60°C did not cause significant changes in lactoferrin and secretory immunoglobulin A, but there was an apparent increase in lysozyme (p = 0.016). There were no significant differences in leptin level among these three forms of milk prior to (p = 0.153) or after heat treatment (p = 0.053). Various freezing/heating/pasteurization processes applied to human milk prior to delivery to neonates could affect the concentration of immunomodulatory proteins, especially lactoferrin, secretory immunoglobulin A, and lysozyme. Leptin was unaffected by the various handling processes tested. Fresh milk was found to be the best food for neonates. Further studies are warranted to evaluate the functional activity of these proteins and their effects on infants' immunological status. Copyright © 2013. Published by Elsevier B.V.

  12. Arabidopsis Intracellular NHX-Type Sodium-Proton Antiporters are Required for Seed Storage Protein Processing.

    Science.gov (United States)

    Ashnest, Joanne R; Huynh, Dung L; Dragwidge, Jonathan M; Ford, Brett A; Gendall, Anthony R

    2015-11-01

    The Arabidopsis intracellular sodium-proton exchanger (NHX) proteins AtNHX5 and AtNHX6 have a well-documented role in plant development, and have been used to improve salt tolerance in a variety of species. Despite evidence that intracellular NHX proteins are important in vacuolar trafficking, the mechanism of this role is poorly understood. Here we show that NHX5 and NHX6 are necessary for processing of the predominant seed storage proteins, and also influence the processing and activity of a vacuolar processing enzyme. Furthermore, we show by yeast two-hybrid and bimolecular fluorescence complementation (BiFC) technology that the C-terminal tail of NHX6 interacts with a component of Retromer, another component of the cell sorting machinery, and that this tail is critical for NHX6 activity. These findings demonstrate that NHX5 and NHX6 are important in processing and activity of vacuolar cargo, and suggest a mechanism by which NHX intracellular (IC)-II antiporters may be involved in subcellular trafficking. © The Author 2015. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  13. Vitellogenin-RNAi and ovariectomy each increase lifespan, increase protein storage, and decrease feeding, but are not additive in grasshoppers.

    Science.gov (United States)

    Tetlak, Alicia G; Burnett, Jacob B; Hahn, Daniel A; Hatle, John D

    2015-12-01

    Reduced reproduction has been shown to increase lifespan in many animals, yet the mechanisms behind this trade-off are unclear. We addressed this question by combining two distinct, direct means of life-extension via reduced reproduction, to test whether they were additive. In the lubber grasshopper, Romalea microptera, ovariectomized (OVX) individuals had a ~20% increase in lifespan and a doubling of storage relative to controls (Sham operated). Similarly, young female grasshoppers treated with RNAi against vitellogenin (the precursor to egg yolk protein) had increased fat body mass and halted ovarian growth. In this study, we compared VgRNAi to two control groups that do not reduce reproduction, namely buffer injection (Buffer) and injection with RNAi against a hexameric storage protein (Hex90RNAi). Each injection treatment was tested with and without ovariectomy. Hence, we tested feeding, storage, and lifespans in six groups: OVX and Buffer, OVX and Hex90RNAi, OVX and VgRNAi, Sham and Buffer, Sham and Hex90RNAi, and Sham and VgRNAi. Ovariectomized grasshoppers and VgRNAi grasshoppers each had similar reductions in feeding (~40%), increases in protein storage in the hemolymph (150-300%), and extensions in lifespan (13-21%). Ovariectomized grasshoppers had higher vitellogenin protein levels than did VgRNAi grasshoppers. Last but not least, when ovariectomy and VgRNAi were applied together, there was no greater effect on feeding, protein storage, or longevity. Hence, feeding regulation, and protein storage in insects, may be conserved components of life-extension via reduced reproduction.

  14. Single Cell Synchrotron FT-IR Microspectroscopy Reveals a Link between Neutral Lipid and Storage Carbohydrate Fluxes in S. cerevisiae

    Science.gov (United States)

    Jamme, Frédéric; Vindigni, Jean-David; Méchin, Valérie; Cherifi, Tamazight; Chardot, Thierry; Froissard, Marine

    2013-01-01

    In most organisms, storage lipids are packaged into specialized structures called lipid droplets. These contain a core of neutral lipids surrounded by a monolayer of phospholipids, and various proteins which vary depending on the species. Hydrophobic structural proteins stabilize the interface between the lipid core and aqueous cellular environment (perilipin family of proteins, apolipoproteins, oleosins). We developed a genetic approach using heterologous expression in Saccharomyces cerevisiae of the Arabidopsis thaliana lipid droplet oleosin and caleosin proteins AtOle1 and AtClo1. These transformed yeasts overaccumulate lipid droplets, leading to a specific increase in storage lipids. The phenotype of these cells was explored using synchrotron FT-IR microspectroscopy to investigate the dynamics of lipid storage and cellular carbon fluxes reflected as changes in spectral fingerprints. Multivariate statistical analysis of the data showed a clear effect on storage carbohydrates and more specifically, a decrease in glycogen in our modified strains. These observations were confirmed by biochemical quantification of the storage carbohydrates glycogen and trehalose. Our results demonstrate that neutral lipid and storage carbohydrate fluxes are tightly connected and co-regulated. PMID:24040242

  15. Single cell synchrotron FT-IR microspectroscopy reveals a link between neutral lipid and storage carbohydrate fluxes in S. cerevisiae.

    Directory of Open Access Journals (Sweden)

    Frédéric Jamme

    Full Text Available In most organisms, storage lipids are packaged into specialized structures called lipid droplets. These contain a core of neutral lipids surrounded by a monolayer of phospholipids, and various proteins which vary depending on the species. Hydrophobic structural proteins stabilize the interface between the lipid core and aqueous cellular environment (perilipin family of proteins, apolipoproteins, oleosins. We developed a genetic approach using heterologous expression in Saccharomyces cerevisiae of the Arabidopsis thaliana lipid droplet oleosin and caleosin proteins AtOle1 and AtClo1. These transformed yeasts overaccumulate lipid droplets, leading to a specific increase in storage lipids. The phenotype of these cells was explored using synchrotron FT-IR microspectroscopy to investigate the dynamics of lipid storage and cellular carbon fluxes reflected as changes in spectral fingerprints. Multivariate statistical analysis of the data showed a clear effect on storage carbohydrates and more specifically, a decrease in glycogen in our modified strains. These observations were confirmed by biochemical quantification of the storage carbohydrates glycogen and trehalose. Our results demonstrate that neutral lipid and storage carbohydrate fluxes are tightly connected and co-regulated.

  16. Lack of Population Structure in Coriander Populations Based on SDS (Seed Storage Protein Page Analysis

    Directory of Open Access Journals (Sweden)

    Gülsüm Yaldiz

    2016-08-01

    Full Text Available Genetic variation is prerequisite for plant breeding. Nothing information existed in the literature for available diversity of Coriander accession in Turkey. Plant breeding activities are negligible in Turkey. So in order to start effective plant breeding program in Turkey, information on the available genetic diversity is viable. Therefore we planned to study the genetic variation and population structure of 29 Coriander accessions by seed storage protein (SDS. SDS analysis elaborated the lack of population structure and genetic bottleneck in the Coriander accessions in Turkey. Based on the results of this study, it was clear that sampling strategy was not appropriate and plant introduction should be made from different sources and diverse genotypes should be used as parents to initialize the effective Turkish Coriander breeding program.

  17. Transcriptome study of storage protein genes of field-grown barley in response to inorganic nitrogen fertilizers

    DEFF Research Database (Denmark)

    Hansen, Michael; Bowra, Steve; Lange, Mette

    2010-01-01

    The storage proteins of barley, in terms of both amino acid profile and quantity, are traits strongly influenced by the amount of nitrogen applied. Given this, we performed a developmental expression analysis of the genes from barley grains grown under field conditions to further our understanding...... profile under different N regimes. Reviewing the expression of the storage protein homologues within the families revealed markedly different temporal profiles; for example, some alleles were expressed very early in development. Furthermore, the differential temporal expression of the homologues suggested...

  18. Antioxidant activities of dioscorin, the storage protein of yam (Dioscorea batatas Decne) tuber.

    Science.gov (United States)

    Hou, W C; Lee, M H; Chen, H J; Liang, W L; Han, C H; Liu, Y W; Lin, Y H

    2001-10-01

    Dioscorin, the storage protein of yam (Dioscorea batatas Decne) tuber (which is different from dioscorine found in tubers of Dioscorea hirsuta), was purified to homogeneity after DE-52 ion exchange column according to the methods of Hou et al. (J. Agric. Food Chem. 1999, 47, 2168-2172). A single band of 32 kDa dioscorin was obtained on a sodium dodecyl sulfate-polyacrylamide gel electrophoresis gel with 2-mercaptoethanol treatment. This purified dioscorin was shown by spectrophotometric method to have scavenging activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical in a pH-dependent manner. There is a positive correlation between scavenging effects against DPPH (8-46%) and amounts of 32 kDa dioscorin (5.97-47.80 nmol) added in Tris-HCl buffer (pH 7.9), which are comparable to those of glutathione at the same concentrations. Using electron paramagnetic resonance (EPR) spectrometry for DPPH radical detection, it was found that the intensities of the EPR signal were decreased by 28.6 and 57 nmol of 32 kDa dioscorin in Tris-HCl buffer (pH 7.9) more than in distilled water compared to controls. EPR spectrometry was also used for hydroxyl radical detection. It was found that 32 kDa dioscorin could capture hydroxyl radical, and the intensities of the EPR signal were significantly decreased dose-dependently by 1.79-14.32 nmol of 32 kDa dioscorin (r = 0.975) compared to the control. It is suggested that 32 kDa dioscorin, the storage protein of yam tuber, may play a role as antioxidant in tubers and may be beneficial for health when people take it as a food additive or consume yam tubers.

  19. Physicochemical and functional properties, microstructure, and storage stability of whey protein/polyvinylpyrrolidone based glue sticks

    Directory of Open Access Journals (Sweden)

    Guorong Wang

    2012-11-01

    Full Text Available A glue stick is comprised of solidified adhesive mounted in a lipstick-like push-up tube. Whey is a byproduct of cheese making. Direct disposal of whey can cause environmental pollution. The objective of this study was to use whey protein isolate (WPI as a natural polymer along with polyvinylpyrrolidone (PVP to develop safe glue sticks. Pre-dissolved WPI solution, PVP, sucrose, 1,2-propanediol (PG, sodium stearate, defoamer, and preservative were mixed and dissolved in water at 90 oC and then molded in push-up tubes. Chemical composition, functional properties (bonding strength, glue setting time, gel hardness, extension/retraction, and spreading properties, microstructure, and storage stability of the prototypes were evaluated in comparison with a commercial control. Results showed that all WPI/PVP prototypes had desirable bonding strength and exhibited faster setting than PVP prototypes and control. WPI could reduce gel hardness and form less compact and rougher structures than that of PVP, but there was no difference in bonding strength. PVP and sucrose could increase the hygroscopicity of glue sticks, thus increasing storage stability. Finally, the optimized prototype GS3 (major components: WPI 8.0%, PVP 12.0%, 1,2-propanediol 10.0%, sucrose 10.0%, and stearic sodium 7.0% had a comparable functionality to the commercial control. Results indicated that whey protein could be used as an adhesive polymer for glue stick formulations, which could be used to bond fiber or cellulose derived substrates such as paper.

  20. Effects of sterilization, packaging, and storage on vitamin C degradation, protein denaturation, and glycation in fortified milks.

    Science.gov (United States)

    Gliguem, H; Birlouez-Aragon, I

    2005-03-01

    Monitoring the nutritional quality of dietetic milk throughout its shelf life is particularly important due to the high susceptibility of some vitamins to oxidation, and the continuous development of the Maillard reaction during storage. The objective of this paper was to evaluate the vitamin C content and protein modification by denaturation and glycation on fortified milk samples (growth milks) destined for 1- to 3-yr-old children. The influences of the sterilization process, formulation, packaging, and storage duration at ambient temperature in the dark were studied. Vitamin C degradation was particularly influenced by type of packaging. The use of a 3-layered opaque bottle was associated with complete oxidation of vitamin C after 1 mo of storage, whereas in the 6-layered opaque bottle, which has an oxygen barrier, the vitamin C content slowly decreased to reach 25% of the initial concentration after 4 mo of storage. However, no significant effect of vitamin C degradation during storage could be observed in terms of Maillard reaction, despite the fact that a probable impact occurred during sterilization. Furosine content and the FAST (fluorescence of advanced Maillard products and soluble tryptophan) index-indicators of the early and advanced Maillard reaction, respectively-were significantly higher in the in-bottle sterilized milk samples compared with UHT samples, and in fortified milk samples compared with cow milk. However, after 1 mo, the impact of storage was predominant, increasing the furosine level and the FAST index at similar levels for the differently processed samples. The early Maillard reaction developed continuously throughout the storage period.In conclusion, only packaging comprising an oxygen and light barrier is compatible with vitamin C fortification of milk. Furthermore, short storage time or low storage temperature is needed to retard vitamin C degradation, protein denaturation, and development of the Maillard reaction.

  1. The purification and characterization of a third storage protein (convicilin) from the seeds of pea (Pisum sativum L.).

    OpenAIRE

    Croy, R R; Gatehouse, J A; Tyler, M; Boulter, D

    1980-01-01

    A third storage protein, distinct from legumin and vicilin, has been purified from the seeds of pea (Pisum sativum L.). This protein has been named 'convicilin' and is present in protein bodies isolated from pea seeds. Convicilin has a subunit mol.wt. of 71 000 and a mol.wt. in its native form of 290 000. Convicilin is antigenically dissimilar to legumin, but gives a reaction of identity with vicilin when tested against antibodies raised against both proteins. However, convicilin contains no ...

  2. OCS in He droplets

    Energy Technology Data Exchange (ETDEWEB)

    Grebenev, V.

    2000-06-01

    Phenomenon of superfluidity of para-hydrogen (pH{sub 2}){sub 1-17} and helium {sup 4}He{sub 1-7000} systems doped with an OCS chromophore molecule was investigated in this work. The study of such systems became possible after the development of the depletion spectroscopy technique in helium droplets. The droplets can be easily created and doped with up to 100 particles such as OCS, para-hydrogen or ortho-hydrogen molecules and {sup 4}He atoms. The measured infrared depletion spectra give the information about the temperature of the droplets and their aggregate state. The depletion spectrum of OCS in pure {sup 4}He droplets was comprehensively studied. The rovibrational OCS spectrum shows well resolved narrow lines. The spectrum is shifted to the red relative to the corresponding gas phase spectrum and the rotational constant of OCS in {sup 4}He droplet is three times smaller than that for free molecule. Different models of OCS rotation in the helium environment were discussed. It was shown that the shapes of the rovibrational lines are defined mainly by inhomogeneous broadening due to the droplet size distribution. The sub-rotational structure of the OCS rovibrational lines was revealed in microwave-infrared double resonance experiments. This structure arises due to the interaction of the OCS with the He environment. However, the information obtained in the experiments was not enough to understand the nature of this interaction. (orig.)

  3. Droplet microfluidics in (bio) chemical analysis

    Czech Academy of Sciences Publication Activity Database

    Basova, E. Y.; Foret, František

    2015-01-01

    Roč. 140, č. 1 (2015), s. 22-38 ISSN 0003-2654 R&D Projects: GA ČR(CZ) GBP206/12/G014 Institutional support: RVO:68081715 Keywords : droplet chemistry * bio analysis * microfluidics * protein Subject RIV: CB - Analytical Chemistry, Separation Impact factor: 4.033, year: 2015

  4. Droplet microfluidics in (bio) chemical analysis

    Czech Academy of Sciences Publication Activity Database

    Basova, E. Y.; Foret, František

    2015-01-01

    Roč. 140, č. 1 (2015), s. 22-38 ISSN 0003-2654 R&D Projects: GA ČR(CZ) GBP206/12/G014 Institutional support: RVO:68081715 Keywords : droplet chemistry * bioanalysis * microfluidics * protein Subject RIV: CB - Analytical Chemistry, Separation Impact factor: 4.033, year: 2015

  5. Recombinant dioscorins of the yam storage protein expressed in Escherichia coli exhibit antioxidant and immunomodulatory activities.

    Science.gov (United States)

    Jheng, Yi-Jyun; Tsai, Wei-Yi; Chen, Kuo-Hsuan; Lin, Kuo-Wei; Chyan, Chia-Lin; Yang, Ching-Chi; Lin, Kuo-Chih

    2012-09-01

    Dioscorins, the major storage proteins in yam tubers, exhibit biochemical and immunomodulatroy activities. To investigate the potential application of dioscorins in biomedical research, we expressed the dioscorin genes Dj-dioA3 and Dp-dioA2 from Dioscorea japonica and Dioscorea pseudojaponica, respectively, in E. coli and routinely obtained approximately 15 mg proteins per liter Escherichia coli culture (mg/L) to 30 mg/L of rDj-dioscorinA3 and 4 to 8 mg/L of rDp-dioscorinA2. Western blot analyses revealed that both recombinant dioscorins contained epitopes with similar antigenicities to those of the native dioscorins. Results from dithiothreitol (DTT) treatment followed by monobromobimane (mBBr) staining showed that both recombinant dioscorins, like the native dioscorins, contain an intramolecular disulfide bond between Cys(28) and Cys(187) residues. Circular dichroism spectroscopy findings indicated that the secondary structural contents of the recombinant dioscorins showed high similarity to those of their corresponding native dioscorins. Both recombinant dioscorins, like the native dioscorins, exhibited 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and Toll-like receptor 4 signaling activities, and stimulated the phagocytosis of E. coli by macrophage. Overall, our results indicated that substantial amounts of recombinant dioscorins can be purified easily from E. coli and that these recombinant dioscorins are appropriate for application in future investigations of the biomedical functions of dioscorins. Copyright © 2012 Elsevier Inc. All rights reserved.

  6. Integrated transcriptomics and proteomics analysis of storage protein composition in developing barley grain to improve nutritional profile

    DEFF Research Database (Denmark)

    Kaczmarczyk, Agnieszka Ewa; Dionisio, Giuseppe; Renaut, Jenny

    2012-01-01

    The aim of the study was to understand the molecular and biochemical mechanisms underpinning the effect of nitrogen (N) on barley (Hordeum vulgare) storage protein production (hordeins) during grain filling. Using a combination of advanced biochemistry methods, we could comprehensively describe c...

  7. Dynamic regulation of hepatic lipid droplet properties by diet.

    Science.gov (United States)

    Crunk, Amanda E; Monks, Jenifer; Murakami, Aya; Jackman, Matthew; Maclean, Paul S; Ladinsky, Mark; Bales, Elise S; Cain, Shannon; Orlicky, David J; McManaman, James L

    2013-01-01

    Cytoplasmic lipid droplets (CLD) are organelle-like structures that function in neutral lipid storage, transport and metabolism through the actions of specific surface-associated proteins. Although diet and metabolism influence hepatic CLD levels, how they affect CLD protein composition is largely unknown. We used non-biased, shotgun, proteomics in combination with metabolic analysis, quantitative immunoblotting, electron microscopy and confocal imaging to define the effects of low- and high-fat diets on CLD properties in fasted-refed mice. We found that the hepatic CLD proteome is distinct from that of CLD from other mammalian tissues, containing enzymes from multiple metabolic pathways. The hepatic CLD proteome is also differentially affected by dietary fat content and hepatic metabolic status. High fat feeding markedly increased the CLD surface density of perilipin-2, a critical regulator of hepatic neutral lipid storage, whereas it reduced CLD levels of betaine-homocysteine S-methyltransferase, an enzyme regulator of homocysteine levels linked to fatty liver disease and hepatocellular carcinoma. Collectively our data demonstrate that the hepatic CLD proteome is enriched in metabolic enzymes, and that it is qualitatively and quantitatively regulated by diet and metabolism. These findings implicate CLD in the regulation of hepatic metabolic processes, and suggest that their properties undergo reorganization in response to hepatic metabolic demands.

  8. Increasing Sucrose Uptake Capacity of Wheat Grains Stimulates Storage Protein Synthesis1[W

    Science.gov (United States)

    Weichert, Nicola; Saalbach, Isolde; Weichert, Heiko; Kohl, Stefan; Erban, Alexander; Kopka, Joachim; Hause, Bettina; Varshney, Alok; Sreenivasulu, Nese; Strickert, Marc; Kumlehn, Jochen; Weschke, Winfriede; Weber, Hans

    2010-01-01

    Increasing grain sink strength by improving assimilate uptake capacity could be a promising approach toward getting higher yield. The barley (Hordeum vulgare) sucrose transporter HvSUT1 (SUT) was expressed under control of the endosperm-specific Hordein B1 promoter (HO). Compared with the wild type, transgenic HOSUT grains take up more sucrose (Suc) in vitro, showing that the transgene is functional. Grain Suc levels are not altered, indicating that Suc fluxes are influenced rather than steady-state levels. HOSUT grains have increased percentages of total nitrogen and prolamins, which is reflected in increased levels of phenylalanine, tyrosine, tryptophan, isoleucine, and leucine at late grain development. Transcript profiling indicates specific stimulation of prolamin gene expression at the onset of storage phase. Changes in gene expression and metabolite levels related to carbon metabolism and amino acid biosynthesis suggest deregulated carbon-nitrogen balance, which together indicate carbon sufficiency and relative depletion of nitrogen. Genes, deregulated together with prolamin genes, might represent candidates, which respond positively to assimilate supply and are related to sugar-starch metabolism, cytokinin and brassinosteroid functions, cell proliferation, and sugar/abscisic acid signaling. Genes showing inverse expression patterns represent potential negative regulators. It is concluded that HvSUT1 overexpression increases grain protein content but also deregulates the metabolic status of wheat (Triticum aestivum) grains, accompanied by up-regulated gene expression of positive and negative regulators related to sugar signaling and assimilate supply. In HOSUT grains, alternating stimulation of positive and negative regulators causes oscillatory patterns of gene expression and highlights the capacity and great flexibility to adjust wheat grain storage metabolism in response to metabolic alterations. PMID:20018590

  9. Ocatin. A novel tuber storage protein from the andean tuber crop oca with antibacterial and antifungal activities.

    Science.gov (United States)

    Flores, Teresita; Alape-Girón, Alberto; Flores-Díaz, Marietta; Flores, Hector E

    2002-04-01

    The most abundant soluble tuber protein from the Andean crop oca (Oxalis tuberosa Mol.), named ocatin, has been purified and characterized. Ocatin accounts for 40% to 60% of the total soluble oca tuber proteins, has an apparent molecular mass of 18 kD and an isoelectric point of 4.8. This protein appears to be found only in tubers and is accumulated only within the cells of the pith and peridermis layers (peel) of the tuber as it develops. Ocatin inhibits the growth of several phytopathogenic bacteria (Agrobacterium tumefaciens, Agrobacterium radiobacter, Serratia marcescens, and Pseudomonas aureofaciens) and fungi (Phytophthora cinnamomi, Fusarium oxysporum, Rhizoctonia solani, and Nectria hematococcus). Ocatin displays substantial amino acid sequence similarity with a widely distributed group of intracellular pathogenesis-related proteins with a hitherto unknown biological function. Our results showed that ocatin serves as a storage protein, has antimicrobial properties, and belongs to the Betv 1/PR-10/MLP protein family. Our findings suggest that an ancient scaffolding protein was recruited in the oca tuber to serve a storage function and that proteins from the Betv 1/PR-10/MLP family might play a role in natural resistance to pathogens.

  10. Ocatin. A Novel Tuber Storage Protein from the Andean Tuber Crop Oca with Antibacterial and Antifungal Activities1

    Science.gov (United States)

    Flores, Teresita; Alape-Girón, Alberto; Flores-Díaz, Marietta; Flores, Hector E.

    2002-01-01

    The most abundant soluble tuber protein from the Andean crop oca (Oxalis tuberosa Mol.), named ocatin, has been purified and characterized. Ocatin accounts for 40% to 60% of the total soluble oca tuber proteins, has an apparent molecular mass of 18 kD and an isoelectric point of 4.8. This protein appears to be found only in tubers and is accumulated only within the cells of the pith and peridermis layers (peel) of the tuber as it develops. Ocatin inhibits the growth of several phytopathogenic bacteria (Agrobacterium tumefaciens, Agrobacterium radiobacter, Serratia marcescens, and Pseudomonas aureofaciens) and fungi (Phytophthora cinnamomi, Fusarium oxysporum, Rhizoctonia solani, and Nectria hematococcus). Ocatin displays substantial amino acid sequence similarity with a widely distributed group of intracellular pathogenesis-related proteins with a hitherto unknown biological function. Our results showed that ocatin serves as a storage protein, has antimicrobial properties, and belongs to the Betv 1/PR-10/MLP protein family. Our findings suggest that an ancient scaffolding protein was recruited in the oca tuber to serve a storage function and that proteins from the Betv 1/PR-10/MLP family might play a role in natural resistance to pathogens. PMID:11950978

  11. Effects of Storage and Granary Weevil Infestation on Gel Electrophoresis and Protein Solubility Properties of Hard and Soft Wheat Flours.

    Science.gov (United States)

    Keskin, Sule; Yalçin, Erkan; Özkaya, Hazim

    2018-02-24

    The objective of this study was to investigate the effects of storage and granary weevil, Sitophilus granarius (L.; Coleoptera: Curculionidae), infestation on pH, protein solubility (PS) and gel electrophoresis properties of meal and roller-milled flours of hard (Ceyhan-99 cv.) and soft (Eser cv.) wheat cultivars, respectively, after 6 mo of storage. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) technique was applied for studying the electrophoretic properties. Hard and soft wheats were infested with non-sexed S. granarius at a rate of two adults/ kg, and stored for 6 mo at 30 ± 1°C and 70 ± 5% RH. The pest-free wheat samples were used as control. The infested and its control samples were collected monthly, and after cleaning the granary weevils, they were hammer-milled or roller-milled in order to get meal flours and roller-milled flours, respectively. The effect of infestation on the storage proteins was more obvious in meal flours than that of the roller-milled flours. Granary weevil feeding resulted secreting of hydrolyzing enzymes and increased the acidity of flours; subsequently the breaking and releasing of some storage proteins generally caused a decrease in pH and an increase in PS values of the meal flours of wheat cultivars. SDS-PAGE results generally indicated that towards the end of storage, the insect population, that greatly increased, caused minor protein depletions resulting decreasing protein band intensities between 113 and 58 kDa of hard wheat meal flour and 101 and 40 kDa of soft wheat roller-milled flour. Consequently, the potential effect of changes probably occurred in high molecular weight glutenin subunits of both wheat cultivars.

  12. Proteomics for exploiting diversity of lupin seed storage proteins and their use as nutraceuticals for health and welfare.

    Science.gov (United States)

    Cabello-Hurtado, Francisco; Keller, Jean; Ley, José; Sanchez-Lucas, Rosa; Jorrín-Novo, Jesús V; Aïnouche, Abdelkader

    2016-06-30

    Lupins have a variety of both traditional and modern uses. In the last decade, reports assessing the benefits of lupin seed proteins have proliferated and, nowadays, the pharmaceutical industry is interested in lupin proteins for human health. Modern genomics and proteomics have hugely contributed to describing the diversity of lupin storage genes and, above all, proteins. Most of these studies have been centered on few edible lupin species. However, Lupinus genus comprises hundreds of species spread throughout the Old and New Worlds, and these resources have been scarcely explored and exploited. We present here a detailed review of the literature on the potential of lupin seed proteins as nutraceuticals, and the use of -omic tools to analyze seed storage polypeptides in main edible lupins and their diversity at the Lupinus inter- and intra-species level. In this sense, proteomics, more than any other, has been a key approach. Proteomics has shown that lupin seed protein diversity, where post-translational modifications yield a large number of peptide variants with a potential concern in bioactivity, goes far beyond gene diversity. The future extended use of second and third generation proteomics should definitely help to go deeper into coverage and characterization of lupin seed proteome. Some important topics concerning storage proteins from lupin seeds are presented and analyzed in an integrated way in this review. Proteomic approaches have been essential in characterizing lupin seed protein diversity, which goes far beyond gene diversity since the protein level adds to the latter differential proteolytic cleavage of conglutin pro-proteins and a diverse array of glycosylation forms and sites. Proteomics has also proved helpful for screening and studying Lupinus germplasm with the future aim of exploiting and improving food production, quality, and nutritional values. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Molecular structure, dynamics and hydration studies of soybean storage proteins and model systems by nuclear magnetic resonance

    International Nuclear Information System (INIS)

    Kakalis, L.T.

    1989-01-01

    The potential of high-resolution 13 C NMR for the characterization of soybean storage proteins was explored. The spectra of a commercial soy protein isolate as well as those of alkali-denatured 7S and 11S soybean globulins were well resolved and tentatively assigned. Relaxation measurements indicated fast motion for several side chains and the protein backbone. Protein fractions (11S and 7S) were also investigated at various states of molecular association. The large size of the multisubunit soybean storage proteins affected adversely both the resolution and the sensitivity of their 13 C NMR spectra. A comparison of 17 O and 2 H NMR relaxation rates of water in solutions of lysozyme (a model system) as a function of concentration, pH and magnetic field suggested that only 17 O monitors directly the hydration of lysozyme. Analysis of 17 O NMR lysozyme hydration data in terms of a two-state, fast-exchange, anisotropic model resulted in hydration parameters which are consistent with the protein's physico-chemical properties. The same model was applied to the calculation of the amount and mobility of bound water in soy protein dispersions by means of 17 O NMR relaxation measurements as a function of protein concentration. The protein concentration dependences of 1 H transverse NMR relaxation measurements at various pH and ionic strength values were fitted by a viral expansion. The interpretation of the data was based on the effects of protein aggregation, salt binding and protein group ionization on the NMR measurements. In all cases, relaxation rates showed a linear dependence on protein activity

  14. Antioxidant activity of pomegranate peel extract on lipid and protein oxidation in beef meatballs during frozen storage.

    Science.gov (United States)

    Turgut, Sebahattin Serhat; Işıkçı, Fatma; Soyer, Ayla

    2017-07-01

    Antioxidant effect of pomegranate peel extract (PE) to retard lipid and protein oxidation in beef meatballs was investigated during frozen storage at -18±1°C. Concentrated and freeze dried aqueous extract of pomegranate peel was incorporated into freshly prepared meatball mix at 0.5% and 1.0% concentrations, and compared with 0.01% butylated hydroxytoluene (BHT) and control (without any antioxidant). In PE treated samples, particularly in high PE concentration, peroxide, malondialdehyde and carbonyl formation, loss of total protein solubility and sulfhydryl groups were significantly lower than control after 6months of storage. A diminution of both myofibrillar (MP) and sarcoplasmic (SP) proteins of high molecular weight was detected after 6months of the storage according to gel electrophoresis patterns. The 1.0% PE led to maintain colour intensity (C) and hue (h°) value. The results from sensory analyses revealed that PE addition to meatballs was effective on preventing rancid odour formation. Addition of both 0.5 and 1% PE in meatballs reduced lipid and protein oxidation and improved sensory scores. These results indicated that PE was effective on retarding lipid and protein oxidations. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Influence of storage and heating on protein glycation levels of processed lactose-free and regular bovine milk products.

    Science.gov (United States)

    Milkovska-Stamenova, Sanja; Hoffmann, Ralf

    2017-04-15

    Thermal treatment preserves the microbiological safety of milk, but also induces Maillard reactions modifying for example proteins. The purpose of this study was evaluating the influence of consumer behaviors (storage and heating) on protein glycation degrees in bovine milk products. Lactosylation and hexosylation sites were identified in ultra-high temperature (UHT), lactose-free pasteurized, and lactose-free UHT milk (ULF) and infant formula (IF) using tandem mass spectrometry (electron transfer dissociation). Overall, 303 lactosylated and 199 hexosylated peptides were identified corresponding to 170 lactosylation (31 proteins) and 117 hexosylation sites (25 proteins). In quantitative terms, storage increased lactosylation up to fourfold in UHT and IF and hexosylation up to elevenfold in ULF and threefold in IF. These levels increased additionally twofold when the stored samples were heated (40°C). In conclusion, storage and heating appear to influence protein glycation levels in milk at similar or even higher degrees than industrial processing. Copyright © 2016 Elsevier Ltd. All rights reserved.

  16. Changes of microbial spoilage, lipid-protein oxidation and physicochemical properties during post mortem refrigerated storage of goat meat.

    Science.gov (United States)

    Sabow, Azad Behnan; Sazili, Awis Qurni; Aghwan, Zeiad Amjad; Zulkifli, Idrus; Goh, Yong Meng; Ab Kadir, Mohd Zainal Abidin; Nakyinsige, Khadijah; Kaka, Ubedullah; Adeyemi, Kazeem Dauda

    2016-06-01

    Examined was the effect of post mortem refrigerated storage on microbial spoilage, lipid-protein oxidation and physicochemical traits of goat meat. Seven Boer bucks were slaughtered, eviscerated and aged for 24 h. The Longissimus lumborum (LL) and Semitendinosus (ST) muscles were excised and subjected to 13 days post mortem refrigerated storage. The pH, lipid and protein oxidation, tenderness, color and drip loss were determined in LL while microbiological analysis was performed on ST. Bacterial counts generally increased with increasing aging time and the limit for fresh meat was reached at day 14 post mortem. Significant differences were observed in malondialdehyde (MDA) content at day 7 of storage. The thiol concentration significantly reduced as aging time increased. The band intensities of myosin heavy chain (MHC) and troponin-T significantly decreased as storage progressed, while actin remained relatively stable. After 14 days of aging, tenderness showed significant improvement while muscle pH and drip loss reduced with increase in storage time. Samples aged for 14 days had higher lightness (P goat meat. © 2016 Japanese Society of Animal Science.

  17. Characterization of the yam tuber storage proteins from Dioscorea batatas exhibiting unique lectin activities.

    Science.gov (United States)

    Gaidamashvili, Mariam; Ohizumi, Yuki; Iijima, Shinichiro; Takayama, Tomo; Ogawa, Tomohisa; Muramoto, Koji

    2004-06-18

    Four major proteins designated DB1, DB2, DB3, and DB4 were isolated and characterized from the yam tuber Dioscorea batatas. The ratios of their yields were 20:50:20:10. DB1 was a mannose-binding lectin (20 kDa) consisting of 10-kDa subunits and was classified as the monocot mannose-binding lectin family. DB2, accounting for 50% of the total protein, was the storage protein, commonly called dioscorins consisting of a 31-kDa subunit. On the basis of amino acid sequence, DB2 was classified to be dioscorin A. DB3 was a maltose-binding lectin, having an apparent molecular mass of 120 kDa and composed of a 66-kDa subunit and two 31-kDa subunits (DB3S). The 66-kDa subunit was further composed of two 31-kDa subunits (DB3L) cross-linked by disulfide bonds. DB3L and DB3S (242 and 241 amino acid residues, respectively) were homologous with each other with 72% sequence identity. They showed a sequence homology to dioscorin B and dioscorin A from Dioscorea alata, with 90 and 93% identity, respectively, and to carbonic anhydrase from Arabidopsis thaliana with about 45% identity. DB3S had one intrachain disulfide bond located at Cys(28)-Cys(187), whereas DB3L had one interchain disulfide bond (Cys(40)-Cys(40)') in addition to the intrachain disulfide bond (Cys(28)-Cys(188)) to form a 66-kDa subunit. DB1 and DB3 agglutinated rabbit erythrocytes at 2.7 and 3.9 microg/ml, respectively. Despite the structural homology between DB2 and DB3, DB2 had no lectin activity. The 66-kDa subunit itself revealed the full hemagglutinating activity of DB3, indicating that DB3L but not DB3S was responsible for the activity. The hemagglutinating activity of DB3 required Ca(2+) ions and was exclusively inhibited by maltose and oligomaltoses (e.g. maltopentaose and maltohexaose) but not by d-glucose. DB3 could not be classified into any known plant lectin family. DB4 was a chitinase, homologous to an acidic chitinase from Dioscorea japonica. DB1, DB2, and DB3 did not show any activity of carbonic

  18. Proteomic Analysis of Differentially Accumulated Proteins in Cucumber (Cucumis sativus) Fruit Peel in Response to Pre-storage Cold Acclimation.

    Science.gov (United States)

    Wang, Bin; Shen, Fei; Zhu, Shijiang

    2017-01-01

    Harvested fruits are still living organs and respond to environmental stimuli. Low temperature storage is effective in extending life of harvested fruit, but it may also cause chilling injury. Cold acclimation has been shown to induce chilling tolerance in plants, but what proteomic changes caused by cold acclimation are related to defense against chilling stress remains largely unclear. Here, 3 d of pre-storage cold acclimation (PsCA) at 10°C reduced chilling injury and secondary disease severity in cucumber stored at 5°C by 51 and 94%, respectively, compared with the control which was directly stored at 5°C. Proteomic analysis of cucumber peel identified 21 significant differentially-accumulated proteins (SDAPs) right after PsCA treatment and 23 after the following cold storage (PsCA+CS). These proteins are mainly related to stress response and defense (SRD), energy metabolism, protein metabolism, signal transduction, primary metabolism, and transcription. The SRD proteins, which made up 37% of the 21 and 47% of the 23, respectively, represented the largest class of SDAPs, and all but one protein were up-regulated, suggesting accumulation of proteins involved in defense response is central feature of proteomic profile changes brought about by PsCA. In fruit just after PsCA treatment, the identified SDAPs are related to responses to various stresses, including chilling, salt stress, dehydration, fungi, bacteria, insects, and DNA damage. However, after prolonged cold storage, the targeted proteins in acclimated fruit were narrowed down in scope to those involved in defense against chilling and pathogens. The change patterns at the transcription level of the majority of the up-regulated differentially-accumulated proteins were highly consistent with those at protein level. Taken all, the results suggest that the short-time cold acclimation initiated comprehensive defense responses in cucumber fruit at first, while the long term storage thereafter altered the

  19. Proteomic Analysis of Differentially Accumulated Proteins in Cucumber (Cucumis sativus Fruit Peel in Response to Pre-storage Cold Acclimation

    Directory of Open Access Journals (Sweden)

    Bin Wang

    2018-01-01

    Full Text Available Harvested fruits are still living organs and respond to environmental stimuli. Low temperature storage is effective in extending life of harvested fruit, but it may also cause chilling injury. Cold acclimation has been shown to induce chilling tolerance in plants, but what proteomic changes caused by cold acclimation are related to defense against chilling stress remains largely unclear. Here, 3 d of pre-storage cold acclimation (PsCA at 10°C reduced chilling injury and secondary disease severity in cucumber stored at 5°C by 51 and 94%, respectively, compared with the control which was directly stored at 5°C. Proteomic analysis of cucumber peel identified 21 significant differentially-accumulated proteins (SDAPs right after PsCA treatment and 23 after the following cold storage (PsCA+CS. These proteins are mainly related to stress response and defense (SRD, energy metabolism, protein metabolism, signal transduction, primary metabolism, and transcription. The SRD proteins, which made up 37% of the 21 and 47% of the 23, respectively, represented the largest class of SDAPs, and all but one protein were up-regulated, suggesting accumulation of proteins involved in defense response is central feature of proteomic profile changes brought about by PsCA. In fruit just after PsCA treatment, the identified SDAPs are related to responses to various stresses, including chilling, salt stress, dehydration, fungi, bacteria, insects, and DNA damage. However, after prolonged cold storage, the targeted proteins in acclimated fruit were narrowed down in scope to those involved in defense against chilling and pathogens. The change patterns at the transcription level of the majority of the up-regulated differentially-accumulated proteins were highly consistent with those at protein level. Taken all, the results suggest that the short-time cold acclimation initiated comprehensive defense responses in cucumber fruit at first, while the long term storage thereafter

  20. Effect of irradiation and soaking in BHT and sodium pyrophosphate on meat proteins and lipids during cold storage

    International Nuclear Information System (INIS)

    Hassan, I.M.; Emam, O.A.

    1988-01-01

    The effect of irradiation treatments up to 10 KGy, soaking in a solution containing 0.5% Na-pyrophosphate and 250 ppm butylated hydroxy toluene (BHT) and a combination of both treatments on the nitrogen content and solubility, protein fractions and lipids stability in beef steaks during cold storage at 4 ± 1°C was followed until the samples were rejected by sensory evaluation. The least effective radiation doses for soluble protein nitrogen (SPN), total soluble nitrogen (TSN) and total nitrogen (TN) were 2, 5 and 10 KGy, respectively. Such effects were proportionally related to the applied dose. The loss in nitrogen compounds and/or their solubility which occurred upon irradiation appeared to be retarded by soaking treatment. Irradiation treatments induced additional protein fraction which seems to be originated from the sarcoplasmic proteins. After the resolution of rigor mortis, the incremental rate of nitrogen extractability was inversely related to the irradiation dose. Another protein fraction was detected only in the 10 KGy irradiated samples after 14 days of cold storage which might be originating from fibrillar proteins as a result of its interaction with some lipid oxidation products. However, soaking treatment itself caused extensive changes in protein fractions, in contrast, protection from radiation and radiation after-effects were observed

  1. Yam storage protein dioscorins from Dioscorea alata and Dioscorea japonica exhibit distinct immunomodulatory activities in mice.

    Science.gov (United States)

    Lin, Pei-Lan; Lin, Kuo-Wei; Weng, Ching-Feng; Lin, Kuo-Chih

    2009-06-10

    The aim of this study was to elucidate the effect of the major storage protein dioscorin isolated from two different yam species, Tainong No. 1 (TN1-dioscorins) and Japanese yam (Dj-dioscorins), on the immune activities of mice. Dj-dioscorins, like TN1-dioscorins, could induce expression of the pro-inflammatory cytokines and stimulate phagocytosis of RAW 264.7. Intraperitoneal injection of the TN1-dioscorins into mice stimulated phagocytosis of bone marrow, spleen, and thymic cells. In contrast, the T and B cells in bone marrow, spleen, and thymus isolated from mice injected with Dj-dioscorins had higher proliferative responses to mitogens. Furthermore, Dj-dioscorins enhanced proliferation of CD4(+), CD8(+), and Tim3(+) (Th1) cells in spleen and CD19(+) cells in both spleen and thymus. Supplement of Dj-dioscorins in the lymphoid cells isolated from Dj-dioscorins primed mice induced cell proliferation of both spleen and thymic cells. These findings indicated that TN1-dioscorins have a higher ability to stimulate the phagocytic activity of the lymphoid cells than Dj-dioscorins, whereas Dj-dioscorins possess more abilities than TN1-dioscorins to enhance the proliferation of the lymphoid cells.

  2. Genetic diversity in radish germplasm for morphological traits and seed storage proteins

    International Nuclear Information System (INIS)

    Jatoi, S.A.; Siddiqui, S.U.; Masood, M.S.; Javaid, A.; Iqbal, M.; Sayal, O.U.

    2011-01-01

    Genetic variation of forty-nine local and exotic radish genotypes including two checks was studied for morphological traits and seed storage protein electrophoresis using sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) markers. A high variation in germplasm for root shape, root length, root colour (internal and external), flesh texture and root type was observed. Among these genotypes, the genetic variation was apparent for most of the characters like plant biomass, root weight, leaf length, root length and root diameter that indicated the potential for crop improvement in these traits through simple selection. Exotic germplasm exhibited higher variation for plant biomass, root weight and root length which could be utilized through breeding programme. Cluster analysis on the basis of genetic diversity for seven quantitative traits resulted into four clusters. No clustering was found on the basis of origin. Low level of variance was observed for SDS-PAGE electrophoresis that suggested acquisition of more germplasm. On the basis of high yield and crispy root texture some genotypes (10076, 10362, 10429, 10658, 10662 and 10667) were identified for further testing under wide range of agro-ecological conditions. (author)

  3. Chip-based droplet sorting

    Energy Technology Data Exchange (ETDEWEB)

    Beer, Neil Reginald; Lee, Abraham; Hatch, Andrew

    2017-11-21

    A non-contact system for sorting monodisperse water-in-oil emulsion droplets in a microfluidic device based on the droplet's contents and their interaction with an applied electromagnetic field or by identification and sorting.

  4. Survival of salmonella transformed to express green fluorescent protein on Italian parsley as affected by processing and storage.

    Science.gov (United States)

    Duffy, E A; Cisneros-Zevallos, L; Castillo, A; Pillai, S D; Ricke, S C; Acuff, G R

    2005-04-01

    To study the effect of processing and storage parameters on the survival of Salmonella on fresh Italian parsley, parsley bunches were dipped for 3 or 15 min in suspensions that were preequilibrated to 5, 25, or 35 degrees C and inoculated with Salmonella transformed to express enhanced green fluorescent protein. Loosely attached and/or associated, strongly attached and/or associated, and internalized and/or entrapped Salmonella cells were enumerated over 0, 1, and 7 days of storage at 25 degrees C and over 0, 1, 7, 14, and 30 days of storage at 4 degrees C using surface-plating procedures. Leaf sections obtained from samples after 0, 1, and 7 days of storage were examined using confocal scanning laser microscopy. Temperature of the dip suspension had little effect on the attachment and survival of Salmonella cells on parsley. Regardless of the temperature or duration of dip, Salmonella was internalized. Immersion for longer times resulted in higher numbers of attached and internalized cells. Microscopic observations supported these results and revealed Salmonella cells near the stomata and within cracks in the cuticle. Storage temperature had the greatest impact on the survival of Salmonella cells on parsley. When stored at 25 degrees C, parsley had a shelf life of 7 days, and Salmonella populations significantly increased over the 7 days of storage. For parsley stored at 4 degrees C, numbers of Salmonella cells decreased over days 0, 1, and 7. After 7 days of storage, there were no viable internalized Salmonella cells detected. Storage temperature represents an important control point for the safety of fresh parsley.

  5. LRRK2 mediated Rab8a phosphorylation promotes lipid storage.

    Science.gov (United States)

    Yu, Miao; Arshad, Muhammad; Wang, Wenmin; Zhao, Dongyu; Xu, Li; Zhou, Linkang

    2018-02-27

    Several mutations in leucine rich repeat kinase 2 (LRRK2) gene have been associated with pathogenesis of Parkinson's disease (PD), a neurodegenerative disorder marked by resting tremors, and rigidity, leading to Postural instability. It has been revealed that mutations that lead to an increase of kinase activity of LRRK2 protein are significantly associated with PD pathogenesis. Recent studies have shown that some Rab GTPases, especially Rab8, serve as substrates of LRRK2 and undergo phosphorylation in its switch II domain upon interaction. Current study was performed in order to find out the effects of the phosphorylation of Rab8 and its mutants on lipid metabolism and lipid droplets growth. The phosphorylation status of Rab8a was checked by phos-tag gel. Point mutant construct were generated to investigate the function of Rab8a. 3T3L1 cells were transfected with indicated plasmids and the lipid droplets were stained with Bodipy. Fluorescent microscopy experiments were performed to examine the sizes of lipid droplets. The interactions between Rab8a and Optineurin were determined by immunoprecipitation and western blot. Our assays demonstrated that Rab8a was phosphorylated by mutated LRRK2 that exhibits high kinase activity. Phosphorylation of Rab8a on amino acid residue T72 promoted the formation of large lipid droplets. T72D mutant of Rab8a had higher activity to promote the formation of large lipid droplets compared with wild type Rab8a, with increase in average diameter of lipid droplets from 2.10 μm to 2.46 μm. Moreover, phosphorylation of Rab8a weakened the interaction with its effector Optineurin. Y1699C mutated LRRK2 was able to phosphorylate Rab8a and phosphorylation of Rab8a on site 72 plays important role in the fusion and enlargement of lipid droplets. Taken together, our study suggests an indirect relationship between enhanced lipid storage capacity and PD pathogenesis.

  6. Hydrodynamics of a quark droplet

    DEFF Research Database (Denmark)

    Bjerrum-Bohr, Johan J.; Mishustin, Igor N.; Døssing, Thomas

    2012-01-01

    We present a simple model of a multi-quark droplet evolution based on the hydrodynamical description. This model includes collective expansion of the droplet, effects of the vacuum pressure and surface tension. The hadron emission from the droplet is described following Weisskopf's statistical...

  7. N-terminomics reveals control of Arabidopsis seed storage proteins and proteases by the Arg/N-end rule pathway.

    Science.gov (United States)

    Zhang, Hongtao; Gannon, Lucy; Hassall, Kirsty L; Deery, Michael J; Gibbs, Daniel J; Holdsworth, Michael J; van der Hoorn, Renier A L; Lilley, Kathryn S; Theodoulou, Frederica L

    2018-05-01

    The N-end rule pathway of targeted protein degradation is an important regulator of diverse processes in plants but detailed knowledge regarding its influence on the proteome is lacking. To investigate the impact of the Arg/N-end rule pathway on the proteome of etiolated seedlings, we used terminal amine isotopic labelling of substrates with tandem mass tags (TMT-TAILS) for relative quantification of N-terminal peptides in prt6, an Arabidopsis thaliana N-end rule mutant lacking the E3 ligase PROTEOLYSIS6 (PRT6). TMT-TAILS identified over 4000 unique N-terminal peptides representing c. 2000 protein groups. Forty-five protein groups exhibited significantly increased N-terminal peptide abundance in prt6 seedlings, including cruciferins, major seed storage proteins, which were regulated by Group VII Ethylene Response Factor (ERFVII) transcription factors, known substrates of PRT6. Mobilisation of endosperm α-cruciferin was delayed in prt6 seedlings. N-termini of several proteases were downregulated in prt6, including RD21A. RD21A transcript, protein and activity levels were downregulated in a largely ERFVII-dependent manner. By contrast, cathepsin B3 protein and activity were upregulated by ERFVIIs independent of transcript. We propose that the PRT6 branch of the pathway regulates protease activities in a complex manner and optimises storage reserve mobilisation in the transition from seed to seedling via control of ERFVII action. © 2017 The Authors. New Phytologist © 2017 New Phytologist Trust.

  8. Selfbound quantum droplets

    Science.gov (United States)

    Langen, Tim; Wenzel, Matthias; Schmitt, Matthias; Boettcher, Fabian; Buehner, Carl; Ferrier-Barbut, Igor; Pfau, Tilman

    2017-04-01

    Self-bound many-body systems are formed through a balance of attractive and repulsive forces and occur in many physical scenarios. Liquid droplets are an example of a self-bound system, formed by a balance of the mutual attractive and repulsive forces that derive from different components of the inter-particle potential. On the basis of the recent finding that an unstable bosonic dipolar gas can be stabilized by a repulsive many-body term, it was predicted that three-dimensional self-bound quantum droplets of magnetic atoms should exist. Here we report on the observation of such droplets using dysprosium atoms, with densities 108 times lower than a helium droplet, in a trap-free levitation field. We find that this dilute magnetic quantum liquid requires a minimum, critical number of atoms, below which the liquid evaporates into an expanding gas as a result of the quantum pressure of the individual constituents. Consequently, around this critical atom number we observe an interaction-driven phase transition between a gas and a self-bound liquid in the quantum degenerate regime with ultracold atoms.

  9. Influence of cooking methods and storage time on lipid and protein oxidation and heterocyclic aromatic amines production in bacon.

    Science.gov (United States)

    Soladoye, O P; Shand, P; Dugan, M E R; Gariépy, C; Aalhus, J L; Estévez, M; Juárez, M

    2017-09-01

    This study aimed to examine the influence of cooking methods and pre-determined refrigerated storage days on the production of lipid oxidation (TBARS), protein oxidation (PROTOX) and heterocyclic aromatic amines (HAA) in bacon. Forty-four pork bellies selected from pigs varying in breed, sex and diets to introduce variability in composition were processed as bacon. Sliced-bacon was stored at 4°C either for 2 or 28days and these storage groups were cooked either with microwave or frying pan. Microwave led to significantly higher PROTOX (P0.05) by the cooking methods and storage times. Similarly, the fatty acid composition of pork belly did not significantly influence the production of HAA, TBARS and PROTOX produced in bacon during cooking. Overall, microwave cooking had lesser impact on the production of carcinogenic compounds in bacon with only minor impact on sensory attributes. Copyright © 2017. Published by Elsevier Ltd.

  10. Analytical detection techniques for droplet microfluidics—A review

    International Nuclear Information System (INIS)

    Zhu, Ying; Fang, Qun

    2013-01-01

    Graphical abstract: -- Highlights: •This is the first review paper focused on the analytical techniques for droplet-based microfluidics. •We summarized the analytical methods used in droplet-based microfluidic systems. •We discussed the advantage and disadvantage of each method through its application. •We also discuss the future development direction of analytical methods for droplet-based microfluidic systems. -- Abstract: In the last decade, droplet-based microfluidics has undergone rapid progress in the fields of single-cell analysis, digital PCR, protein crystallization and high throughput screening. It has been proved to be a promising platform for performing chemical and biological experiments with ultra-small volumes (picoliter to nanoliter) and ultra-high throughput. The ability to analyze the content in droplet qualitatively and quantitatively is playing an increasing role in the development and application of droplet-based microfluidic systems. In this review, we summarized the analytical detection techniques used in droplet systems and discussed the advantage and disadvantage of each technique through its application. The analytical techniques mentioned in this paper include bright-field microscopy, fluorescence microscopy, laser induced fluorescence, Raman spectroscopy, electrochemistry, capillary electrophoresis, mass spectrometry, nuclear magnetic resonance spectroscopy, absorption detection, chemiluminescence, and sample pretreatment techniques. The importance of analytical detection techniques in enabling new applications is highlighted. We also discuss the future development direction of analytical detection techniques for droplet-based microfluidic systems

  11. Raman imaging and lipidomic analysis of lipid droplets in (activated) hepatic stellate cells

    NARCIS (Netherlands)

    Vaandrager, A.B.; Testerink, N.; Ajat, M.; Houweling, M.; Brouwers, J.F.H.M.; Pully, V.V.; van Manen, H.J.; Otto, Cornelis; Helms, J.B.

    2009-01-01

    In eukaryotic cells the excess of hydrophobic molecules is stored in special organelles named lipid droplets (LDs). These droplets contain triacylglycerides, cholesteryl esters and/or retinyl esters, depending on the function of the cell in which they reside. Retinyl esters, the storage form of

  12. Efficacy of various protein-based coating on enhancing the shelf life of fresh eggs during storage.

    Science.gov (United States)

    Caner, Cengiz; Yüceer, Muhammed

    2015-07-01

    The effectiveness of various coatings (whey protein isolate [WPI], whey protein concentrate [WPC], zein, and shellac) on functional properties, interior quality, and eggshell breaking strength of fresh eggs were evaluated during storage at 24 °: C for 6 weeks. Coatings and storage time had significant effects on Haugh unit, yolk index, albumen pH, dry matter (DMA), relative whipping capacity (RWC), and albumen viscosity. Uncoated eggs had higher albumen pH (9.56) and weight loss, and lower albumen viscosity (5.73), Haugh unit (HU), and yolk index (YI) during storage. Among the coated eggs, the shellac and zein coated eggs had the highest value of albumen viscosity (27.26 to 26.90), HU (74.10 to 73.61), and YI (44.84 to 44.63) after storage. Shellac (1.44%) was more effective in preventing weight loss than WPC (4.59%), WPI (4.60%), and zein (2.13%) coatings. Uncoated eggs had the higest value (6.71%) of weight lost. All coatings increased shell strength (5.18 to 5.73 for top and 3.58 to 4.71 for bottom) significantly (P eggs (4.70 for top and 3.15 for bottom). The functional properties such as albumen DMA (14.50 to 16.66 and 18.97 for uncoated) and albumen RWC (841 to 891 and 475 for uncoated) of fresh eggs can be preserved during storage when they are coated. The shellac and zein coatings were more effective for maintaining the internal quality of fresh eggs during storage. Fourier transform near infrared (FT-NIR) in the 800 to 2500 nm reflection spectra were used to quantify the contents of the fresh eggs at the end of storage. Eggs coated with shellac or zein displayed a higher absorbance at 970 and 1,197 nm respectively (OH vibration of water) compared with those coated with WPI or WPC and the uncoated group at the end of storage. The coatings improved functional properties and also shell strength and could be a viable alternative technology for maintaining the internal quality of eggs during long-term storage. This study highlights the promising use of

  13. Biophysical evaluation of aminoclay as an effective protectant for protein stabilization during freeze-drying and storage

    Directory of Open Access Journals (Sweden)

    Song JG

    2016-12-01

    Full Text Available Jae Geun Song, Sang Hoon Lee, Hyo-Kyung Han College of Pharmacy, Dongguk University, Goyang, South Korea Abstract: This study aimed to evaluate aminoclay (3-aminopropyl-functionalized magnesium phyllosilicate as an effective protectant for the stabilization of protein formulation in freeze-drying. Bovine serum albumin (BSA, as a model protein, was freeze-dried with aminoclay at various concentrations, and the effects of aminoclay on the structural stability of proteins were compared with those of the conventional stabilizers. The structural characteristics of the protein were determined by size exclusion chromatography (SEC, circular dichroism (CD, and Fourier transform infrared (FTIR spectroscopy. Furthermore, physicochemical and morphological characteristics were examined by X-ray powder diffraction (XRPD, differential scanning calorimetry (DSC, and scanning electron microscopy (SEM. XRPD and DSC patterns indicated that the glass transition temperature (Tg of the amorphous formulation of aminoclay mixed with proteins was gradually elevated as the concentration of aminoclay increased. FTIR and CD spectral analysis suggested that the protein structure was well maintained with aminoclay during the freeze-drying process and 3 months of storage at 4°C and 40°C. Furthermore, aminoclay conferred the greatest protection against aggregation and retained the monomer content of BSA even at a high temperature. The morphological characteristics of lyophilized proteins were also well conserved during the storage with aminoclay. These results suggested that aminoclay may be useful as an alternative stabilizer for maintaining the structural stability of protein formulations. Keywords: aminoclay, cryoprotectant, lyoprotectant, freeze-drying, protein, stability

  14. A functional genomics approach to understand the control and regulation of storage protein biosynthesis in barley grain

    DEFF Research Database (Denmark)

    Vincze, É; Hansen, M; Bowra, S

    2008-01-01

    assembled in our laboratory. To identify coregulated genes, a distance matrix was constructed and we identified three clusters corresponding to the early, middle and late seed development. The gene expression pattern associated with the clusters was investigated using pathway specific analysis with specific......The aim of the study was to obtain an insight into amino acid and storage protein metabolism in the developing barley grain at the molecular level. Our strategy was to analyse the transcriptome of relevant pathways in developing grains of field grown barley using a grain specific microarray...... pathways in the barley grain. The study described here could provide a strong complement to existing knowledge assisting further  understanding of seed development and thereby provide a foundation for plant breeding towards storage protein with improved nutritional quality....

  15. A study of different indicators of Maillard reaction with whey proteins and different carbohydrates under adverse storage conditions.

    Science.gov (United States)

    Leiva, Graciela E; Naranjo, Gabriela B; Malec, Laura S

    2017-01-15

    This study examined different indicators of each stage of Maillard reaction under adverse storage conditions in a system with whey proteins and lactose or glucose. The analysis of lysine loss by the o-phthaldialdehyde method can be considered a good indicator of the early stage, showing considerable differences in reactivity when systems with mono and disaccharides were analyzed. Capillary electrophoresis proved to be a sensitive method for evaluating the extent of glycosylation of the native proteins, providing valuable information when the loss of lysine was not significant. The estimation of the Amadori compound from the determination of total 5-hydroxymethyl-2-furfuraldehyde would have correlate well with reactive lysine content if the advanced stages of the reaction had not been reached. For assessing the occurrence of the intermediate and final stages, the measurement of free 5-hydroxymethyl-2-furfuraldehyde and color, proved not to be suitable for storage conditions. Copyright © 2016 Elsevier Ltd. All rights reserved.

  16. Coregulation of Soybean Vegetative Storage Protein Gene Expression by Methyl Jasmonate and Soluble Sugars 1

    Science.gov (United States)

    Mason, Hugh S.; DeWald, Daryll B.; Creelman, Robert A.; Mullet, John E.

    1992-01-01

    The soybean vegetative storage protein genes vspA and vspB are highly expressed in developing leaves, stems, flowers, and pods as compared with roots, seeds, and mature leaves and stems. In this paper, we report that physiological levels of methyl jasmonate (MeJA) and soluble sugars synergistically stimulate accumulation of vsp mRNAs. Treatment of excised mature soybean (Glycine max Merr. cv Williams) leaves with 0.2 molar sucrose and 10 micromolar MeJA caused a large accumulation of vsp mRNAs, whereas little accumulation occurred when these compounds were supplied separately. In soybean cell suspension cultures, the synergistic effect of sucrose and MeJA on the accumulation of vspB mRNA was maximal at 58 millimolar sucrose and was observed with fructose or glucose substituted for sucrose. In dark-grown soybean seedlings, the highest levels of vsp mRNAs occurred in the hypocotyl hook, which also contained high levels of MeJA and soluble sugars. Lower levels of vsp mRNAs, MeJA, and soluble sugars were found in the cotyledons, roots, and nongrowing regions of the stem. Wounding of mature soybean leaves induced a large accumulation of vsp mRNAs when wounded plants were incubated in the light. Wounded plants kept in the dark or illuminated plants sprayed with dichlorophenyldimethylurea, an inhibitor of photosynthetic electron transport, showed a greatly reduced accumulation of vsp mRNAs. The time courses for the accumulation of vsp mRNAs induced by wounding or sucrose/MeJA treatment were similar. These results strongly suggest that vsp expression is coregulated by endogenous levels of MeJA (or jasmonic acid) and soluble carbohydrate during normal vegetative development and in wounded leaves. ImagesFigure 1Figure 4Figure 5 PMID:16668757

  17. Immunomodulatory activity of dioscorin, the storage protein of yam (Dioscorea alata cv. Tainong No. 1) tuber.

    Science.gov (United States)

    Liu, Yen-Wenn; Shang, Huey-Fang; Wang, Chung-Kwe; Hsu, Feng-Lin; Hou, Wen-Chi

    2007-11-01

    The purified dioscorin from yam (Dioscorea alata L. cv. Tainong 1) tuber was previously reported (Hsu et al., 2002. J. Agric. Food Chem., 50, 6109-6113). In this report, we evaluated its immunomodulatory ability in vitro in the presence of polymyxin B (50 microg/ml) to eliminate lipopolysaccharide (LPS) contamination. Dioscorin (5-100 microg/ml) was able to stimulate nitric oxide production (expressed as nitrite concentrations) in RAW264.7 cells. The stimulation index on the phagocytosis of RAW264.7 cells against E. coli and the oxidative burst (determined by the intensity of rhodamine fluorescence) of RAW264.7 cells were both enhanced by different concentrations of dioscorin (5-100 microg/ml). The cytokine production, including IL-6, TNF-alpha, and IL-1beta in dioscorin-treated RAW264.7 cells or human monocytes, was measured in the cultured medium. Dioscorin (5-100 microg/ml) was found able to induce IL-6, TNF-alpha, and IL-1beta production in RAW264.7 cells and human monocytes. To evaluate the effects of dioscorin on the proliferation of spleen cells from BALB/c mice, phytohemagglutinin (PHA, 2 microg/ml) alone or PHA mixed with different concentrations of dioscorin (10, 25, and 50 microg/ml) was used to treat spleen cells for 24h. The stimulated proliferation index of splenic cells ranged from 1.38- to 1.48-fold of PHA alone for PHA mixed with different concentrations of dioscorin (10, 25, and 50 microg/ml). We suggest that the tuber storage protein of yam dioscorin functions as an immunomodulatory substance.

  18. Coregulation of soybean vegetative storage protein gene expression by methyl jasmonate and soluble sugars.

    Science.gov (United States)

    Mason, H S; Dewald, D B; Creelman, R A; Mullet, J E

    1992-03-01

    The soybean vegetative storage protein genes vspA and vspB are highly expressed in developing leaves, stems, flowers, and pods as compared with roots, seeds, and mature leaves and stems. In this paper, we report that physiological levels of methyl jasmonate (MeJA) and soluble sugars synergistically stimulate accumulation of vsp mRNAs. Treatment of excised mature soybean (Glycine max Merr. cv Williams) leaves with 0.2 molar sucrose and 10 micromolar MeJA caused a large accumulation of vsp mRNAs, whereas little accumulation occurred when these compounds were supplied separately. In soybean cell suspension cultures, the synergistic effect of sucrose and MeJA on the accumulation of vspB mRNA was maximal at 58 millimolar sucrose and was observed with fructose or glucose substituted for sucrose. In dark-grown soybean seedlings, the highest levels of vsp mRNAs occurred in the hypocotyl hook, which also contained high levels of MeJA and soluble sugars. Lower levels of vsp mRNAs, MeJA, and soluble sugars were found in the cotyledons, roots, and nongrowing regions of the stem. Wounding of mature soybean leaves induced a large accumulation of vsp mRNAs when wounded plants were incubated in the light. Wounded plants kept in the dark or illuminated plants sprayed with dichlorophenyldimethylurea, an inhibitor of photosynthetic electron transport, showed a greatly reduced accumulation of vsp mRNAs. The time courses for the accumulation of vsp mRNAs induced by wounding or sucrose/MeJA treatment were similar. These results strongly suggest that vsp expression is coregulated by endogenous levels of MeJA (or jasmonic acid) and soluble carbohydrate during normal vegetative development and in wounded leaves.

  19. Electrical actuation of dielectric droplets

    International Nuclear Information System (INIS)

    Kumari, N; Bahadur, V; Garimella, S V

    2008-01-01

    Electrical actuation of liquid droplets at the microscale offers promising applications in the fields of microfluidics and lab-on-a-chip devices. Much prior research has targeted the electrical actuation of electrically conducting liquid droplets; however, the actuation of dielectric droplets has remained relatively unexplored, despite the advantages associated with the use of a dielectric droplet. This paper presents modeling and experimental results on the electrical actuation of dielectric droplets between two flat plates. A first-order analytical model, based on the energy-minimization principle, is developed to estimate the electrical actuation force on a dielectric droplet as it moves between two flat plates. Two versions of this analytical model are benchmarked for their suitability and accuracy against a detailed numerical model. The actuation force prediction is then combined with available semi-analytical expressions for predicting the forces opposing droplet motion to develop a model that predicts transient droplet motion under electrical actuation. Electrical actuation of dielectric droplets is experimentally demonstrated by moving transformer oil droplets between two flat plates under the influence of an actuation voltage. Droplet velocities and their dependence on the plate spacing and the applied voltage are experimentally measured and showed reasonable agreement with predictions from the models developed

  20. Viability and growth promotion of starter and probiotic bacteria in yogurt supplemented with whey protein hydrolysate during refrigerated storage

    Directory of Open Access Journals (Sweden)

    Anna Dąbrowska

    2017-11-01

    Full Text Available The effect of whey protein hydrolysate (WPH addition on growth of standard yoghurt cultures and Bifidobacterium adolescentis during co-fermentation and its viability during storage at 4ºC in yoghurts has been evaluated. WPH was obtained with the use of serine protease from Y. lipolytica yeast. Stirred probiotic yoghurts were prepared by using whole milk standardized to 16% of dry matter with the addition of either whey protein concentrate, skim milk powder (SMP, WPH-SMP (ratio 1:1, WPH. The hydrolysate increased the yoghurt culture counts at the initial stage of fermentation and significantly inhibited the decrease in population viability throughout the storage at 4ºC in comparison to the control. The post-fermentation acidification was also retarded by the addition of WPH. The hydrolysate did not increase the Bifidobacterium adolescentis counts at the initial stage. However, the WPH significantly improved its viability. After 21 days of storage, in the yogurts supplemented with WPH, the population of these bacteria oscillated around 3.04 log10 CFU/g, while in samples where SMP or whey protein concentrate was used, the bacteria were no longer detected.

  1. Viability and growth promotion of starter and probiotic bacteria in yogurt supplemented with whey protein hydrolysate during refrigerated storage.

    Science.gov (United States)

    Dąbrowska, Anna; Babij, Konrad; Szołtysik, Marek; Chrzanowska, Józefa

    2017-11-22

    The effect of whey protein hydrolysate (WPH) addition on growth of standard yoghurt cultures and Bifidobacterium adolescentis during co-fermentation and its viability during storage at 4ºC in yoghurts has been evaluated. WPH was obtained with the use of serine protease from Y. lipolytica yeast. Stirred probiotic yoghurts were prepared by using whole milk standardized to 16% of dry matter with the addition of either whey protein concentrate, skim milk powder (SMP), WPH-SMP (ratio 1:1), WPH. The hydrolysate increased the yoghurt culture counts at the initial stage of fermentation and significantly inhibited the decrease in population viability throughout the storage at 4ºC in comparison to the control. The post-fermentation acidification was also retarded by the addition of WPH. The hydrolysate did not increase the Bifidobacterium adolescentis counts at the initial stage. However, the WPH significantly improved its viability. After 21 days of storage, in the yogurts supplemented with WPH, the population of these bacteria oscillated around 3.04 log10 CFU/g, while in samples where SMP or whey protein concentrate was used, the bacteria were no longer detected.

  2. Diel pattern of circadian clock and storage protein gene expression in leaves and during seed filling in cowpea (Vigna unguiculata).

    Science.gov (United States)

    Weiss, Julia; Terry, Marta I; Martos-Fuentes, Marina; Letourneux, Lisa; Ruiz-Hernández, Victoria; Fernández, Juan A; Egea-Cortines, Marcos

    2018-02-14

    Cowpea (Vigna unguiculata) is an important source of protein supply for animal and human nutrition. The major storage globulins VICILIN and LEGUMIN (LEG) are synthesized from several genes including LEGA, LEGB, LEGJ and CVC (CONVICILIN). The current hypothesis is that the plant circadian core clock genes are conserved in a wide array of species and that primary metabolism is to a large extent controlled by the plant circadian clock. Our aim was to investigate a possible link between gene expression of storage proteins and the circadian clock. We identified cowpea orthologues of the core clock genes VunLHY, VunTOC1, VunGI and VunELF3, the protein storage genes VunLEG, VunLEGJ, and VunCVC as well as nine candidate reference genes used in RT-PCR. ELONGATION FACTOR 1-A (ELF1A) resulted the most suitable reference gene. The clock genes VunELF3, VunGI, VunTOC1 and VunLHY showed a rhythmic expression profile in leaves with a typical evening/night and morning/midday phased expression. The diel patterns were not completely robust and only VungGI and VungELF3 retained a rhythmic pattern under free running conditions of darkness. Under field conditions, rhythmicity and phasing apparently faded during early pod and seed development and was regained in ripening pods for VunTOC1 and VunLHY. Mature seeds showed a rhythmic expression of VunGI resembling leaf tissue under controlled growth chamber conditions. Comparing time windows during developmental stages we found that VunCVC and VunLEG were significantly down regulated during the night in mature pods as compared to intermediate ripe pods, while changes in seeds were non-significant due to high variance. The rhythmic expression under field conditions was lost under growth chamber conditions. The core clock gene network is conserved in cowpea leaves showing a robust diel expression pattern except VunELF3 under growth chamber conditions. There appears to be a clock transcriptional reprogramming in pods and seeds compared to

  3. Droplets and sprays

    CERN Document Server

    Sazhin, Sergei

    2014-01-01

    Providing a clear and systematic description of droplets and spray dynamic models, this book maximises reader insight into the underlying physics of the processes involved, outlines the development of new physical and mathematical models, and broadens understanding of interactions between the complex physical processes which take place in sprays. Complementing approaches based on the direct application of computational fluid dynamics (CFD), Droplets and Sprays treats both theoretical and practical aspects of internal combustion engine process such as the direct injection of liquid fuel, subcritical heating and evaporation. Includes case studies that illustrate the approaches relevance to automotive applications,  it is also anticipated that the described models can find use in other areas such as in medicine and environmental science.

  4. Performance of droplet generator and droplet collector in liquid droplet radiator under microgravity

    Science.gov (United States)

    Totani, T.; Itami, M.; Nagata, H.; Kudo, I.; Iwasaki, A.; Hosokawa, S.

    2002-06-01

    The Liquid Droplet Radiator (LDR) has an advantage over comparable conventional radiators in terms of the rejected heat power-weight ratio. Therefore, the LDR has attracted attention as an advanced radiator for high-power space systems that will be prerequisite for large space structures. The performance of the LDR under microgravity condition has been studied from the viewpoint of operational space use of the LDR in the future. In this study, the performances of a droplet generator and a droplet collector in the LDR are investigated using drop shafts in Japan: MGLAB and JAMIC. As a result, it is considered that (1) the droplet generator can produce uniform droplet streams in the droplet diameter range from 200 to 280 [µm] and the spacing range from 400 to 950 [µm] under microgravity condition, (2) the droplet collector with the incidence angle of 35 degrees can prevent a uniform droplet stream, in which droplet diameter is 250 [µm] and the velocity is 16 [m/s], from splashing under microgravity condition, whereas splashes may occur at the surface of the droplet collector in the event that a nonuniform droplet stream collides against it.

  5. New droplet model developments

    International Nuclear Information System (INIS)

    Dorso, C.O.; Myers, W.D.; Swiatecki, W.J.; Moeller, P.; Treiner, J.; Weiss, M.S.

    1985-09-01

    A brief summary is given of three recent contributions to the development of the Droplet Model. The first concerns the electric dipole moment induced in octupole deformed nuclei by the Coulomb redistribution. The second concerns a study of squeezing in nuclei and the third is a study of the improved predictive power of the model when an empirical ''exponential'' term is included. 25 refs., 3 figs

  6. Acoustic droplet vaporization of vascular droplets in gas embolotherapy

    Science.gov (United States)

    Bull, Joseph

    2016-11-01

    This work is primarily motivated by a developmental gas embolotherapy technique for cancer treatment. In this methodology, infarction of tumors is induced by selectively formed vascular gas bubbles that arise from the acoustic vaporization of vascular droplets. Additionally, micro- or nano-droplets may be used as vehicles for localized drug delivery, with or without flow occlusion. In this talk, we examine the dynamics of acoustic droplet vaporization through experiments and theoretical/computational fluid mechanics models, and investigate the bioeffects of acoustic droplet vaporization on endothelial cells and in vivo. Functionalized droplets that are targeted to tumor vasculature are examined. The influence of fluid mechanical and acoustic parameters, as well as droplet functionalization, is explored. This work was supported by NIH Grant R01EB006476.

  7. Studies on RBC lipid and protein phosphorylation during blood bank storage

    International Nuclear Information System (INIS)

    Dumaswala, U.J.; Bryan, D.J.; Greenwalt, T.J.

    1986-01-01

    Recent evidence has suggested that phosphoinositides play a significant role in maintaining membrane structure and function. Their importance during blood bank storage is not understood. They have performed preliminary studies of the phosphoinositide synthetic pathway enzymes of RBC during blood bank storage. At 0 and 35 days of storage leaky ghosts were prepared and incubated with [γ- 32 P]ATP for 5 minutes at 30 C. One aliquot was subjected to acidified solvent extraction and thin layer chromatography. The labeled phosphoinositide -4,5 biphosphate (PIP 2 ), phosphoinositide-4 phosphate (PIP) and phosphatidic acid (PA) spots were scraped and counted by liquid scintillation spectrometry. Another aliquot was used for SDS-PAGE and the radioactivity associated with the β-spectrin was measured. These experiments suggest a decrease in RBC phosphoinositol and PIP-Kinases and β-spectrin kinase activities during blood bank storage. Further studies are being done to evaluate significance of these observations

  8. Optical calorimetry in microfluidic droplets.

    Science.gov (United States)

    Chamoun, Jacob; Pattekar, Ashish; Afshinmanesh, Farzaneh; Martini, Joerg; Recht, Michael I

    2018-05-29

    A novel microfluidic calorimeter that measures the enthalpy change of reactions occurring in 100 μm diameter aqueous droplets in fluoropolymer oil has been developed. The aqueous reactants flow into a microfluidic droplet generation chip in separate fluidic channels, limiting contact between the streams until immediately before they form the droplet. The diffusion-driven mixing of reactants is predominantly restricted to within the droplet. The temperature change in droplets due to the heat of reaction is measured optically by recording the reflectance spectra of encapsulated thermochromic liquid crystals (TLC) that are added to one of the reactant streams. As the droplets travel through the channel, the spectral characteristics of the TLC represent the internal temperature, allowing optical measurement with a precision of ≈6 mK. The microfluidic chip and all fluids are temperature controlled, and the reaction heat within droplets raises their temperature until thermal diffusion dissipates the heat into the surrounding oil and chip walls. Position resolved optical temperature measurement of the droplets allows calculation of the heat of reaction by analyzing the droplet temperature profile over time. Channel dimensions, droplet generation rate, droplet size, reactant stream flows and oil flow rate are carefully balanced to provide rapid diffusional mixing of reactants compared to thermal diffusion, while avoiding thermal "quenching" due to contact between the droplets and the chip walls. Compared to conventional microcalorimetry, which has been used in this work to provide reference measurements, this new continuous flow droplet calorimeter has the potential to perform titrations ≈1000-fold faster while using ≈400-fold less reactants per titration.

  9. Investigation of total seed storage proteins of pakistani and japanese maize (zea mays l.) through sds-page markers

    International Nuclear Information System (INIS)

    Shinwari, Z.K.

    2014-01-01

    The assessment of genetic diversity among the members of a species is of vital importance for successful breeding and adaptability. In the present study 83 genotypes of maize of Pakistani and Japanese origin were evaluated for the total seed storage proteins using sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) through vertical slab unit. The total protein subunits were separated on 12% polyacrylamide gel using standard protocols. A total of 18 protein subunits were noted out of which 7 (39%) were monomorphic and 11 (61%) were polymorphic, with molecular weight ranging from 10 to 122 kDa. Coefficients of similarity among the accessions ranged between 0.89 and 1.00. The dendrogram obtained through UPGMA clustering method showed two main clusters: 1 and 2. First cluster comprised of 9 genotypes including Sahiwal-2002, while second cluster contained 74 genotypes including Aaiti-2002 and Sadaf. Over all a low level of polymorphism was observed in total seed storage protein patterns of maize genotypes from Pakistan as well as Japan. It is inferred from the present study that more genotypes of maize could be brought under study and more advanced biochemical techniques with more reliable results could be followed to bring assessment of genetic diversity of maize for planning breeding programs. (author)

  10. Properties and regulation of biosynthesis of cottonseed storage proteins. Comprehensive progress report, December 1, 1976 to September 1, 1979

    Energy Technology Data Exchange (ETDEWEB)

    Dure, III, L S

    1979-01-01

    The regulation of gene expression in cotton seed embryogenesis was studied by attempting to define what gene products are likely to be highly regulated during this developmental progression. The flow of nitrogen into the free amino acids pools of the developing cotyledons, and into the principal nitrogen nutritional reserve of the seed, the storage proteins was measured. This was continued by following the flow of nitrogen from the storage proteins to the principal exported amino acid asparagine that occurs during the first several days of germination. In this fashion the rise and fall of certain enzymes of amino acid intermediary metabolism could be postulated, and in some cases, verified. The subsets of abundant mRNAs whose appearance and disappearance coincided with developmental events in cotyledon embryogenesis/germination with the short range goal of identifying proteins/enzyme activities were delineated as well as their mRNAs that represent specific developmental stages and the long range goal of using these representatives as probes for studying the mechanisms controlling the rise and fall of these mRNAs and their protein products.

  11. Evaporation of inclined water droplets

    Science.gov (United States)

    Kim, Jin Young; Hwang, In Gyu; Weon, Byung Mook

    2017-01-01

    When a drop is placed on a flat substrate tilted at an inclined angle, it can be deformed by gravity and its initial contact angle divides into front and rear contact angles by inclination. Here we study on evaporation dynamics of a pure water droplet on a flat solid substrate by controlling substrate inclination and measuring mass and volume changes of an evaporating droplet with time. We find that complete evaporation time of an inclined droplet becomes longer as gravitational influence by inclination becomes stronger. The gravity itself does not change the evaporation dynamics directly, whereas the gravity-induced droplet deformation increases the difference between front and rear angles, which quickens the onset of depinning and consequently reduces the contact radius. This result makes the evaporation rate of an inclined droplet to be slow. This finding would be important to improve understanding on evaporation dynamics of inclined droplets. PMID:28205642

  12. COMPARATIVE DYNAMICS OF PROTEIN DESTRUCTION IN CANNED FOODS IN SAUCE AT DIFFERENT THERMAL TREATMENT REGIMES AND SUBSEQUENT STORAGE

    Directory of Open Access Journals (Sweden)

    V. B. Krylova

    2017-01-01

    Full Text Available In the course of investigations, the structural changes in proteins were established, which were associated with the preliminary treatment of meat ingredients, a pH level of the system and parameters of thermal treatment.The pasteurization regimes allowed retaining a protein nitrogen proportion up to 94% by the end of canned food storage duration. Upon sterilization, the losses in protein nitrogen were two times higher. A negative effect of more acidic sauce on preservation of the protein nitrogen fraction in canned foods was established.An accumulation of the peptide nitrogen fraction in the canned foods in tomato sauce aſter pasteurization was two times more intensive. In the sterilized canned foods, the processes of accumulation of the low molecular weight nitrogenous compounds were more intensive, which suggests a depth of destruction of the protein and peptide nitrogen fraction. It was shown that an accumulation of amino-ammonia nitrogen during canned food storage was on average 12.4% irrespective of the pH value in the used sauces and the type of thermal treatment.A shiſt in the pH value of the canned foods toward the acid side upon pasteurization was noticed. With that, a degree of the shiſt in the canned foods in tomato sauce was 2.5 times higher than the pH value of the canned foods in sour cream sauce. When sterilizing canned foods, another dynamics of the pH values was observed: a pH value declined by 0.39 units in the canned foods in tomato sauce and grew by 0.22 units in the canned foods in sour cream sauce. During storage, the tendency of more intense pH decline was revealed for the canned foods in tomato sauce aſter pasteurization compared to the canned foods aſter sterilization. Another character of the pH value dynamics was found in the canned foods in sour cream sauce: an insignificant increase (by 0.7% of the pH value in the pasteurized canned foods and a significant decrease (by 8.4% in the sterilized canned foods

  13. Droplet generation during core reflood

    International Nuclear Information System (INIS)

    Kocamustafaogullari, G.; De Jarlais, G.; Ishii, M.

    1983-01-01

    The process of entrainment and disintegration of liquid droplets by a flow of steam has considerable practical importance in calculating the effectivenes of the emergency core cooling system. Liquid entrainment is also important in determination of the critical heat flux point in general. Thus the analysis of the reflooding phase of a LOCA requires detailed knowledge of droplet size. Droplet size is mainly determined by the droplet generation mechanisms involved. To study these mechanisms, data generated in the PWR FLECHT SEASET series of experiments was analyzed. In addition, an experiment was performed in which the hydrodynamics of low quality post-CHF flow (inverted annular flow) were simulated in an adiabatic test section

  14. Dioscorin, the major tuber storage protein of yam (Dioscorea batatas decne) with carbonic anhydrase and trypsin inhibitor activities.

    Science.gov (United States)

    Hou, W C; Liu, J S; Chen, H J; Chen, T E; Chang, C F; Lin, Y H

    1999-05-01

    Dioscorin, the tuber storage protein of yam (Dioscorea batatas Decne), was purified successively by ammonium sulfate fractionation, DE-52 ion exchange chromatography, and Sephadex G-75 column. Two protein bands (82 and 28 kDa) were found under nonreducing conditions after SDS-PAGE; but only one band (32 kDa) was detected under reducing conditions. The first 21 amino acids in the N-terminal region of the 28 kDa form were VEDEFSYIEGNPNGPENWGNL, which was highly homologous to deductive sequence of dioscorin from cDNA of another yam species (Dioscoreacayenensis Lam) reported by Conlan et al. (Plant Mol. Biol. 1995, 28, 369-380). Hewett-Emmett and Tashian (Mol. Phylogenet. Evol. 1996, 5, 50 -77) mentioned that, according to DNA alignments, dioscorin from yam (D. cayenensis) was alpha-carbonic anhydrase (alpha-CA) related. In this report, we found that the purified dioscorin showed both CA dehydration activity using sodium bicarbonate as a substrate and CA activity staining after SDS-PAGE. A polyclonal antibody, which was raised against trypsin inhibitor (TI), a storage protein of sweet potato (Ipomoea batatas [L.] Lam var. Tainong 57), cross-reacted with dioscorin, which also showed TI activity determined by both activity staining after SDS-PAGE and trypsin inhibition determination.

  15. Effects of abscisic acid and high osmoticum on storage protein gene expression in microspore embryos of Brassica napus

    International Nuclear Information System (INIS)

    Wilen, R.W.; Mandel, R.M.; Pharis, R.P.; Moloney, M.M.; Holbrook, L.A.

    1990-01-01

    Storage protein gene expression, characteristic of mid- to late embryogenesis, was investigated in microspore embryos of rapeseed (Brassica napus). These embryos, derived from the immature male gametophyte, accumulate little or no detectable napin or cruciferin mRNA when cultured on hormone-free medium containing 13% sucrose. The addition of abscisic acid (ABA) to the medium results in an increase in detectable transcripts encoding both these polypeptides. Storage protein mRNA is induced at 1 micromolar ABA with maximum stimulation occurring between 5 and 50 micromolar. This hormone induction results in a level of storage protein mRNA that is comparable to that observed in zygotic embryos of an equivalent morphological stage. Effects similar to that of ABA are noted when 12.5% sorbitol is added to the microspore embryo medium (osmotic potential = 25.5 bars). Time course experiments, to study the induction of napin and cruciferin gene expression demonstrated that the ABA effect occurred much more rapidly than the high osmoticum effect, although after 48 hours, the levels of napin or cruciferin mRNA detected were similar in both treatments. This difference in the rates of induction is consistent with the idea that the osmotic effect may be mediated by ABA which is synthesized in response to the reduced water potential. Measurements of ABA (by gas chromatography-mass spectrometry using [ 2 H 6 ]ABA as an internal standard) present in microspore embryos during sorbitol treatment and in embryos treated with 10 micromolar ABA were performed to investigate this possibility. Within 2 hours of culture on high osmoticum the level of ABA increased substantially and significantly above control and reached a maximum concentration within 24 hours. This elevated concentration was maintained for 48 hours after culturing and represents a sixfold increase over control embryos

  16. Droplet size effects on film drainage between droplet and substrate.

    Science.gov (United States)

    Steinhaus, Benjamin; Spicer, Patrick T; Shen, Amy Q

    2006-06-06

    When a droplet approaches a solid surface, the thin liquid film between the droplet and the surface drains until an instability forms and then ruptures. In this study, we utilize microfluidics to investigate the effects of film thickness on the time to film rupture for water droplets in a flowing continuous phase of silicone oil deposited on solid poly(dimethylsiloxane) (PDMS) surfaces. The water droplets ranged in size from millimeters to micrometers, resulting in estimated values of the film thickness at rupture ranging from 600 nm down to 6 nm. The Stefan-Reynolds equation is used to model film drainage beneath both millimeter- and micrometer-scale droplets. For millimeter-scale droplets, the experimental and analytical film rupture times agree well, whereas large differences are observed for micrometer-scale droplets. We speculate that the differences in the micrometer-scale data result from the increases in the local thin film viscosity due to confinement-induced molecular structure changes in the silicone oil. A modified Stefan-Reynolds equation is used to account for the increased thin film viscosity of the micrometer-scale droplet drainage case.

  17. Isolation and characterisation of cDNA clones representing the genes encoding the major tuber storage protein (dioscorin) of yam (Dioscorea cayenensis Lam.).

    Science.gov (United States)

    Conlan, R S; Griffiths, L A; Napier, J A; Shewry, P R; Mantell, S; Ainsworth, C

    1995-06-01

    cDNA clones encoding dioscorins, the major tuber storage proteins (M(r) 32,000) of yam (Dioscorea cayenesis) have been isolated. Two classes of clone (A and B, based on hybrid release translation product sizes and nucleotide sequence differences) which are 84.1% similar in their protein coding regions, were identified. The protein encoded by the open reading frame of the class A cDNA insert is of M(r) 30,015. The difference in observed and calculated molecular mass might be attributed to glycosylation. Nucleotide sequencing and in vitro transcription/translation suggest that the class A dioscorin proteins are synthesised with signal peptides of 18 amino acid residues which are cleaved from the mature peptide. The class A and class B proteins are 69.6% similar with respect to each other, but show no sequence identity with other plant proteins or with the major tuber storage proteins of potato (patatin) or sweet potato (sporamin). Storage protein gene expression was restricted to developing tubers and was not induced by growth conditions known to induce expression of tuber storage protein genes in other plant species. The codon usage of the dioscorin genes suggests that the Dioscoreaceae are more closely related to dicotyledonous than to monocotyledonous plants.

  18. Genetic mapping and validation of the loci controlling 7S α' and 11S A-type storage protein subunits in soybean [Glycine max (L.) Merr.].

    Science.gov (United States)

    Boehm, Jeffrey D; Nguyen, Vi; Tashiro, Rebecca M; Anderson, Dale; Shi, Chun; Wu, Xiaoguang; Woodrow, Lorna; Yu, Kangfu; Cui, Yuhai; Li, Zenglu

    2018-03-01

    Four soybean storage protein subunit QTLs were mapped using bulked segregant analysis and an F 2 population, which were validated with an F 5 RIL population. The storage protein globulins β-conglycinin (7S subunit) and glycinin (11S subunits) can affect the quantity and quality of proteins found in soybean seeds and account for more than 70% of the total soybean protein. Manipulating the storage protein subunits to enhance soymeal nutrition and for desirable tofu manufacturing characteristics are two end-use quality goals in soybean breeding programs. To aid in developing soybean cultivars with desired seed composition, an F 2 mapping population (n = 448) and an F 5 RIL population (n = 180) were developed by crossing high protein cultivar 'Harovinton' with the breeding line SQ97-0263_3-1a, which lacks the 7S α', 11S A 1 , 11S A 2 , 11S A 3 and 11S A 4 subunits. The storage protein composition of each individual in the F 2 and F 5 populations were profiled using SDS-PAGE. Based on the presence/absence of the subunits, genomic DNA bulks were formed among the F 2 plants to identify genomic regions controlling the 7S α' and 11S protein subunits. By utilizing polymorphic SNPs between the bulks characterized with Illumina SoySNP50K iSelect BeadChips at targeted genomic regions, KASP assays were designed and used to map QTLs causing the loss of the subunits. Soybean storage protein QTLs were identified on Chromosome 3 (11S A 1 ), Chromosome 10 (7S α' and 11S A 4 ), and Chromosome 13 (11S A 3 ), which were also validated in the F 5 RIL population. The results of this research could allow for the deployment of marker-assisted selection for desired storage protein subunits by screening breeding populations using the SNPs linked with the subunits of interest.

  19. A storage-protein marker associated with the suppressor of Pm8 for powdery mildew resistance in wheat.

    Science.gov (United States)

    Ren, S X; McIntosh, R A; Sharp, P J; The, T T

    1996-11-01

    A suppressor of resistance to powdery mildew conferred by Pm8 showed complete association with the presence of a storage-protein marker resolved by electrophoresis on SDS-PAGE gels. This marker was identified as the product of the gliadin allele Gli-A1a. The mildewresponse phenotypes of wheats possessing the 1BL.1RS translocation were completely predictable from electrophoretograms. The suppressor, designated SuPm8, was located on chromosome 1AS. It was specific in its suppression of Pm8, and did not affect the rye-derived resistance phenotypes of wheat lines with Pm17, also located in 1RS, or of lines with Pm7.

  20. Analysis of iron storage proteins in chicken liver and spleen tissues in comparison with human liver ferritin by Moessbauer spectroscopy

    International Nuclear Information System (INIS)

    Oshtrakh, M.I.; Milder, O.B.; Semionkin, V.A.; Malakheeva, L.I.; Prokopenko, P.G.

    2006-01-01

    Characterization of iron storage proteins in liver and spleen from normal chicken and chicken with lymphoid leukemia in comparison with human liver ferritin were considered by Moessbauer spectroscopy (preliminary results). Small differences in Moessbauer hyperfine parameters for both normal and lymphoid leukemia chicken liver and spleen were observed. The value of quadrupole splitting for human liver ferritin was higher than those for chicken tissues. A decrease of iron content in lymphoid leukemia chicken tissues was also found, however, the reason of this fact (pathology or feeding) was not clear yet. (author)

  1. Zinc deficiency-induced iron accumulation, a consequence of alterations in iron regulatory protein-binding activity, iron transporters, and iron storage proteins.

    Science.gov (United States)

    Niles, Brad J; Clegg, Michael S; Hanna, Lynn A; Chou, Susan S; Momma, Tony Y; Hong, Heeok; Keen, Carl L

    2008-02-22

    One consequence of zinc deficiency is an elevation in cell and tissue iron concentrations. To examine the mechanism(s) underlying this phenomenon, Swiss 3T3 cells were cultured in zinc-deficient (D, 0.5 microM zinc), zinc-supplemented (S, 50 microM zinc), or control (C, 4 microM zinc) media. After 24 h of culture, cells in the D group were characterized by a 50% decrease in intracellular zinc and a 35% increase in intracellular iron relative to cells in the S and C groups. The increase in cellular iron was associated with increased transferrin receptor 1 protein and mRNA levels and increased ferritin light chain expression. The divalent metal transporter 1(+)iron-responsive element isoform mRNA was decreased during zinc deficiency-induced iron accumulation. Examination of zinc-deficient cells revealed increased binding of iron regulatory protein 2 (IRP2) and decreased binding of IRP1 to a consensus iron-responsive element. The increased IRP2-binding activity in zinc-deficient cells coincided with an increased level of IRP2 protein. The accumulation of IRP2 protein was independent of zinc deficiency-induced intracellular nitric oxide production but was attenuated by the addition of the antioxidant N-acetylcysteine or ascorbate to the D medium. These data support the concept that zinc deficiency can result in alterations in iron transporter, storage, and regulatory proteins, which facilitate iron accumulation.

  2. Proteomic analysis of lipid droplets from Caco-2/TC7 enterocytes identifies novel modulators of lipid secretion.

    Directory of Open Access Journals (Sweden)

    Frauke Beilstein

    Full Text Available In enterocytes, the dynamic accumulation and depletion of triacylglycerol (TAG in lipid droplets (LD during fat absorption suggests that cytosolic LD-associated TAG contribute to TAG-rich lipoprotein (TRL production. To get insight into the mechanisms controlling the storage/secretion balance of TAG, we used as a tool hepatitis C virus core protein, which localizes onto LDs, and thus may modify their protein coat and decrease TRL secretion. We compared the proteome of LD fractions isolated from Caco-2/TC7 enterocytes expressing or not hepatitis C virus core protein by a differential proteomic approach (isobaric tag for relative and absolute quantitation (iTRAQ labeling coupled with liquid chromatography and tandem mass spectrometry. We identified 42 proteins, 21 being involved in lipid metabolism. Perilipin-2/ADRP, which is suggested to stabilize long term-stored TAG, was enriched in LD fractions isolated from Caco-2/TC7 expressing core protein while perilipin-3/TIP47, which is involved in LD synthesis from newly synthesized TAG, was decreased. Endoplasmic reticulum-associated proteins were strongly decreased, suggesting reduced interactions between LD and endoplasmic reticulum, where TRL assembly occurs. For the first time, we show that 17β-hydroxysteroid dehydrogenase 2 (DHB2, which catalyzes the conversion of 17-keto to 17 β-hydroxysteroids and which was the most highly enriched protein in core expressing cells, is localized to LD and interferes with TAG secretion, probably through its capacity to inactivate testosterone. Overall, we identified potential new players of lipid droplet dynamics, which may be involved in the balance between lipid storage and secretion, and may be altered in enterocytes in pathological conditions such as insulin resistance, type II diabetes and obesity.

  3. Particle Manipulation Methods in Droplet Microfluidics.

    Science.gov (United States)

    Tenje, Maria; Fornell, Anna; Ohlin, Mathias; Nilsson, Johan

    2018-02-06

    This Feature describes the different particle manipulation techniques available in the droplet microfluidics toolbox to handle particles encapsulated inside droplets and to manipulate whole droplets. We address the advantages and disadvantages of the different techniques to guide new users.

  4. Influence of pre-cooking protein paste gelation conditions and post-cooking gel storage conditions on gel texture.

    Science.gov (United States)

    Paker, Ilgin; Matak, Kristen E

    2016-01-15

    Gelation conditions affect the setting of myofibrillar fish protein gels. Therefore the impact of widely applied pre-cooking gelation time/temperature strategies and post-cooking period on the texture and color of final protein gels was determined. Four pre-cooking gelation strategies (no setting time, 30 min at 25 °C, 1 h at 40 °C or 24 h at 4 °C) were applied to protein pastes (fish protein concentrate and standard functional additives). After cooking, texture and color were analyzed either directly or after 24 h at 4 °C on gels adjusted to 25 °C. No-set gels were harder, gummier and chewier (P cooking. Gel-setting conditions had a greater (P cooking stored gels in texture and color, depending on the pre-cooking gelation strategy. Pre-cooking gelation conditions will affect final protein gel texture and color, with gel stability benefiting from a gel-setting period. However, post-cooking storage may have a greater impact on final gels, with textural attributes becoming more consistent between all samples. © 2015 Society of Chemical Industry.

  5. Vaporization of irradiated droplets

    International Nuclear Information System (INIS)

    Armstrong, R.L.; O'Rourke, P.J.; Zardecki, A.

    1986-01-01

    The vaporization of a spherically symmetric liquid droplet subject to a high-intensity laser flux is investigated on the basis of a hydrodynamic description of the system composed of the vapor and ambient gas. In the limit of the convective vaporization, the boundary conditions at the fluid--gas interface are formulated by using the notion of a Knudsen layer in which translational equilibrium is established. This leads to approximate jump conditions at the interface. For homogeneous energy deposition, the hydrodynamic equations are solved numerically with the aid of the CON1D computer code (''CON1D: A computer program for calculating spherically symmetric droplet combustion,'' Los Alamos National Laboratory Report No. LA-10269-MS, December, 1984), based on the implict continuous--fluid Eulerian (ICE) [J. Comput. Phys. 8, 197 (1971)] and arbitrary Lagrangian--Eulerian (ALE) [J. Comput. Phys. 14, 1227 (1974)] numerical mehtods. The solutions exhibit the existence of two shock waves propagating in opposite directions with respect to the contact discontinuity surface that separates the ambient gas and vapor

  6. Explosive Leidenfrost droplets

    Science.gov (United States)

    Colinet, Pierre; Moreau, Florian; Dorbolo, Stéphane

    2017-11-01

    We show that Leidenfrost droplets made of an aqueous solution of surfactant undergo a violent explosion in a wide range of initial volumes and concentrations. This unexpected behavior turns out to be triggered by the formation of a gel-like shell, followed by a sharp temperature increase. Comparing a simple model of the radial surfactant distribution inside a spherical droplet with experiments allows highlighting the existence of a critical surface concentration for the shell to form. The temperature rise (attributed to boiling point elevation with surface concentration) is a key feature leading to the explosion, instead of the implosion (buckling) scenario reported by other authors. Indeed, under some conditions, this temperature increase is shown to be sufficient to trigger nucleation and growth of vapor bubbles in the highly superheated liquid bulk, stretching the surrounding elastic shell up to its rupture limit. The successive timescales characterizing this explosion sequence are also discussed. Funding sources: F.R.S. - FNRS (ODILE and DITRASOL projects, RD and SRA positions of P. Colinet and S. Dorbolo), BELSPO (IAP 7/38 MicroMAST project).

  7. Protein mobilities and P-selectin storage in Weibel–Palade bodies

    OpenAIRE

    Kiskin, Nikolai I.; Hellen, Nicola; Babich, Victor; Hewlett, Lindsay; Knipe, Laura; Hannah, Matthew J.; Carter, Tom

    2010-01-01

    Using fluorescence recovery after photobleaching (FRAP) we measured the mobilities of EGFP-tagged soluble secretory proteins in the endoplasmic reticulum (ER) and in individual Weibel–Palade bodies (WPBs) at early (immature) and late (mature) stages in their biogenesis. Membrane proteins (P-selectin, CD63, Rab27a) were also studied in individual WPBs. In the ER, soluble secretory proteins were mobile; however, following insertion into immature WPBs larger molecules (VWF, Proregion, tPA) and P...

  8. Disruption of the Arabidopsis CGI-58 homologue produces Chanarin–Dorfman-like lipid droplet accumulation in plants

    OpenAIRE

    James, Christopher N.; Horn, Patrick J.; Case, Charlene R.; Gidda, Satinder K.; Zhang, Daiyuan; Mullen, Robert T.; Dyer, John M.; Anderson, Richard G. W.; Chapman, Kent D.

    2010-01-01

    CGI-58 is the defective gene in the human neutral lipid storage disease called Chanarin-Dorfman syndrome. This disorder causes intracellular lipid droplets to accumulate in nonadipose tissues, such as skin and blood cells. Here, disruption of the homologous CGI-58 gene in Arabidopsis thaliana resulted in the accumulation of neutral lipid droplets in mature leaves. Mass spectroscopy of isolated lipid droplets from cgi-58 loss-of-function mutants showed they contain triacylglycerols with common...

  9. Millifluidic droplet analyser for microbiology

    NARCIS (Netherlands)

    Baraban, L.; Bertholle, F.; Salverda, M.L.M.; Bremond, N.; Panizza, P.; Baudry, J.; Visser, de J.A.G.M.; Bibette, J.

    2011-01-01

    We present a novel millifluidic droplet analyser (MDA) for precisely monitoring the dynamics of microbial populations over multiple generations in numerous (=103) aqueous emulsion droplets (100 nL). As a first application, we measure the growth rate of a bacterial strain and determine the minimal

  10. Performance of starter in yogurt supplemented with soy protein isolate and biotransformation of isoflavones during storage period.

    Science.gov (United States)

    Pham, T Thuy; Shah, Nagendra P

    2009-01-01

    In this study, soy protein isolate (SPI) (4%, v/w) was supplemented to the yogurt mix to increase the amount of biologically active isoflavone in yogurt (SY). The control yogurt was without any SPI supplementation (USY). The supplementation significantly (P yogurt starter including Lactobacillus delbrueckii ssp. bulgaricus ATCC 11842 (Lb 11842) and Streptococcus thermophilus ST 1342 (ST 1342) during the fermentation process by 4.7%. The starter produced more acetic acid and less lactic acid in SY than that in USY and altered the ratio of lactic and acetic acid during the entire storage period. The viability of both Lb 11842 and ST 1342 in SY was significantly (P < 0.05) lower than that in USY from 14 d of the storage period, however, their concentration still remained high (8.11 to 8.84 log CFU/g). The starter transformed 72.8% of total inactive isoflavone glycosides (IG) to active isoflavone aglycones (IA), increasing the IA content from 1.35 to 15.01 mg/100 g sample. During the storage period, IA concentration slowly rose from 15.02 to 15.51 mg/100 g sample.

  11. Proteomic analysis of seed storage proteins in wild rice species of the Oryza genus.

    Science.gov (United States)

    Jiang, Chunmiao; Cheng, Zaiquan; Zhang, Cheng; Yu, Tengqiong; Zhong, Qiaofang; Shen, J Qingxi; Huang, Xingqi

    2014-01-01

    The total protein contents of rice seeds are significantly higher in the three wild rice species (Oryza rufipogon Grill., Oryza officinalis Wall. and Oryza meyeriana Baill.) than in the cultivated rice (Oryza sativa L.). However, there is still no report regarding a systematic proteomic analysis of seed proteins in the wild rice species. Also, the relationship between the contents of seed total proteins and rice nutritional quality has not been thoroughly investigated. The total seed protein contents, especially the glutelin contents, of the three wild rice species were higher than those of the two cultivated rice materials. Based on the protein banding patterns of SDS-PAGE, O. rufipogon was similar to the two cultivated rice materials, followed by O. officinalis, while O. meyeriana exhibited notable differences. Interestingly, O. meyeriana had high contents of glutelin and low contents of prolamine, and lacked 26 kDa globulin band and appeared a new 28 kDa protein band. However, for O. officinali a 16 kDa protein band was absent and a row of unique 32 kDa proteins appeared. In addition, we found that 13 kDa prolamine band disappeared while special 14 kDa and 12 kDa protein bands were present in O. officinalis. Two-dimensional gel electrophoresis (2-DE) analysis revealed remarkable differences in protein profiles of the wild rice species and the two cultivated rice materials. Also, the numbers of detected protein spots of the three wild rice species were significantly higher than those of two cultivated rice. A total of 35 differential protein spots were found for glutelin acidic subunits, glutelin precursors and glutelin basic subunits in wild rice species. Among those, 18 protein spots were specific and 17 major spots were elevated. Six differential protein spots for glutelin acidic subunits were identified, including a glutelin type-A 2 precursor and five hypothetical proteins. This was the first report on proteomic analysis of the three wild rice species

  12. Leidenfrost boiling of water droplet

    Directory of Open Access Journals (Sweden)

    Orzechowski Tadeusz

    2017-01-01

    Full Text Available The investigations concerned a large water droplet at the heating surface temperature above the Leidenfrost point. The heating cylinder was the main component of experimental stand on which investigations were performed. The measurement system was placed on the high-sensitivity scales. Data transmission was performed through RS232 interface. The author-designed program, with extended functions to control the system, was applied. The present paper examines the behaviour of a large single drop levitating over a hot surface, unsteady mass of the drop, and heat transfer. In computations, the dependence, available in the literature, for the orthogonal droplet projection on the heating surface as a function of time was employed. It was confirmed that the local value of the heat transfer coefficient is a power function of the area of the droplet surface projection. Also, a linear relationship between the flux of mass evaporated from the droplet and the droplet orthogonal projection was observed.

  13. Leidenfrost boiling of water droplet

    Science.gov (United States)

    Orzechowski, Tadeusz

    The investigations concerned a large water droplet at the heating surface temperature above the Leidenfrost point. The heating cylinder was the main component of experimental stand on which investigations were performed. The measurement system was placed on the high-sensitivity scales. Data transmission was performed through RS232 interface. The author-designed program, with extended functions to control the system, was applied. The present paper examines the behaviour of a large single drop levitating over a hot surface, unsteady mass of the drop, and heat transfer. In computations, the dependence, available in the literature, for the orthogonal droplet projection on the heating surface as a function of time was employed. It was confirmed that the local value of the heat transfer coefficient is a power function of the area of the droplet surface projection. Also, a linear relationship between the flux of mass evaporated from the droplet and the droplet orthogonal projection was observed.

  14. Liquid droplet radiator technology issues

    International Nuclear Information System (INIS)

    Mattick, A.T.; Hertzberg, A.

    1985-01-01

    The operation of the liquid droplet radiator (LDR) is analyzed to establish design constraints for the LDR components and to predict the performance of an integrated LDR system. The design constraints largely result from mass loss considerations: fluid choice is governed by evaporation loss; droplet generation techniques must be capable of precise aiming of >10 5 droplet streams; and collection losses must be less than 1 droplet in 10 7 . Concepts for droplet generation and collection components are discussed and incorporated into a mass model for an LDR system. This model predicts that LDR's using lithium, Dow 705 silicone fluid, or NaK may be several times lighter than heat pipe radiators. 13 refs

  15. seed storage proteins arl2 and its variants from the apa locus of wild ...

    African Journals Online (AJOL)

    sion of arcelin and ARL2 tepary bean proteins. Furthermore, a reduction in size and weight of emerged adult insects to almost half was observed. This work demonstrates the superior resistance common bean backcross lines to A. obtectus conferred by the presence of the APA proteins introgressed from tepary bean.

  16. Effects of brown seaweed polyphenols, α-tocopherol, and ascorbic acid on protein oxidation and textural properties of fish mince (Pagrosomus major) during frozen storage.

    Science.gov (United States)

    Wang, Tiantian; Li, Zhenxing; Yuan, Fangzhou; Lin, Hong; Pavase, Tushar Ramesh

    2017-03-01

    Frozen storage of minced fish is currently one of the most important techniques to maintain its functional properties. However, some deterioration does occur during frozen storage and cause quality loss. The effects of brown seaweed polyphenols, α-tocopherol, and ascorbic acid on lipid and protein oxidation and textural properties of red sea bream (Pagrosomus major) during 90 days of frozen storage at -18 °C were investigated. All added antioxidants at 1 g kg -1 resulted in significantly lower thiobarbituric acid-reactive substances (TBARS) compared to the control during the 45 days of frozen storage. The antioxidants were also effective in retarding protein oxidation concerning to total sulfhydryl content and protein carbonyl content. Brown seaweed polyphenols and α-tocopherol significantly retarded the inactivation of Ca 2+ -ATPase activity during the first 45 days, whereas ascorbic acid had no such effect. The antioxidant activity showed either an invariable or decrease trend after 45 days storage. Adding antioxidants had a significant effect on the breaking force of the gels during the frozen storage period. These results indicate that brown seaweed polyphenols and α-tocopherol can be used to prevent oxidative reactions and thus maintain the structure of the gel formed by fish mince during frozen storage. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  17. Live Cell Imaging During Germination Reveals Dynamic Tubular Structures Derived from Protein Storage Vacuoles of Barley Aleurone Cells

    Directory of Open Access Journals (Sweden)

    Verena Ibl

    2014-09-01

    Full Text Available The germination of cereal seeds is a rapid developmental process in which the endomembrane system undergoes a series of dynamic morphological changes to mobilize storage compounds. The changing ultrastructure of protein storage vacuoles (PSVs in the cells of the aleurone layer has been investigated in the past, but generally this involved inferences drawn from static pictures representing different developmental stages. We used live cell imaging in transgenic barley plants expressing a TIP3-GFP fusion protein as a fluorescent PSV marker to follow in real time the spatially and temporally regulated remodeling and reshaping of PSVs during germination. During late-stage germination, we observed thin, tubular structures extending from PSVs in an actin-dependent manner. No extensions were detected following the disruption of actin microfilaments, while microtubules did not appear to be involved in the process. The previously-undetected tubular PSV structures were characterized by complex movements, fusion events and a dynamic morphology. Their function during germination remains unknown, but might be related to the transport of solutes and metabolites.

  18. Protein aggregation in aqueous casein solution. Effect of irradiation, dose level, concentration, storage and additives (carbohydrate and lipid)

    Energy Technology Data Exchange (ETDEWEB)

    Yousri, R M

    1980-06-01

    From the vast amount of research efforts dealing with various aspects of radiation effects on foods and food components, it is apparent up to now that much remains to be studied in depth, much may have to be added or corrected about radiation-induced physico-chemical changes in foods. A great many reactions that take place when foodstuffs are subjected to ionizing radiation are still not fully understood. The better understanding of some of the radiation-induced changes in pure proteins as such or in mixture with other food constituents could yield much data which could be meaningfully extrapolated to intact foods and consequently could help to improve the assessment of the wholesomeness of irradiated foods. It was the purpose of our investigations to elucidate some of the changes in the chemical structure of a pure protein (casein), irradiated as such or with added carbohydrate and/or lipid. The effect of subsequent storage of the irradiated solutions has been also examined. The formation of protein aggregates was studied by gel filtration technique. The application of thin-layer gel filtration, its speed and adaptability to very small samples facilitated the measurements of the extent of aggregation which occurred in protein molecules after irradiation.

  19. Influence of sodium nitrite on protein oxidation and nitrosation of sausages subjected to processing and storage.

    Science.gov (United States)

    Feng, Xianchao; Li, Chenyi; Jia, Xu; Guo, Yan; Lei, Na; Hackman, Robert M; Chen, Lin; Zhou, Guanghong

    2016-06-01

    The influence of NaNO2 content on protein oxidation and nitrosation was investigated in cooked sausages at different concentrations (0, 50, 100, 200 and 400 mg NaNO2/kg). Dependent on concentration, NaNO2 had both anti- and pro-oxidant effects on protein oxidation. The antioxidant effects of NaNO2 on the protein oxidation were evidenced by significantly lower carbonyl contents, higher free amines and lower surface hydrophobicities. The pro-oxidant effects of NaNO2 on protein oxidation resulted in a decrease of sulfhydryls and an increase of disulfide bonds. NaNO2 also improved the protein nitrosation inducing the formation of 3-nitrotyrosine (3-NT). Moreover, 3-NT had significant correlations with parameters of protein oxidation, indicating that 3-NT may be a possible marker for protein oxidation. Results of this study contribute to an understanding of the impact of NaNO2 on food quality and help to identify optimal formulations of cured meat products. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. Lossless droplet transfer of droplet-based microfluidic analysis

    Science.gov (United States)

    Kelly, Ryan T [West Richland, WA; Tang, Keqi [Richland, WA; Page, Jason S [Kennewick, WA; Smith, Richard D [Richland, WA

    2011-11-22

    A transfer structure for droplet-based microfluidic analysis is characterized by a first conduit containing a first stream having at least one immiscible droplet of aqueous material and a second conduit containing a second stream comprising an aqueous fluid. The interface between the first conduit and the second conduit can define a plurality of apertures, wherein the apertures are sized to prevent exchange of the first and second streams between conduits while allowing lossless transfer of droplets from the first conduit to the second conduit through contact between the first and second streams.

  1. SDS-Page Seed Storage Protein Profiles in Chili Peppers (Capsicum L.

    Directory of Open Access Journals (Sweden)

    Owk ANIEL KUMAR

    2010-09-01

    Full Text Available Seed protein banding patterns (SDS-PAGE were studied from eighteen genotypes of chili pepper (Capsicum L. A total of 21 protein polypeptide bands with molecular weight ranging from 18.6 to 72.0 kD were recorded. Among the genotypes CA18, CA21 and CA27 represented maximum number of protein bands (12. Band no. (11 and (5,12 are exclusive to C. annuum L. and C. frutescens L. genotypes respectively. Average percent similarity was highest (100% between CA2 and CA8 genotypes and the UPGMA dendrogram represented low genetic diversity. The study revealed that considerable intra and inter-specific differences were found in the genotypes. The variability of protein profiles in the genotypes suggested that these selected genotypes can be a good source for crop improvement through hybridization programs.

  2. Influence of Prolonged Storage Process, Pasteurization, and Heat Treatment on Biologically-active Human Milk Proteins

    Directory of Open Access Journals (Sweden)

    Jih-Chin Chang

    2013-12-01

    Conclusion: Various freezing/heating/pasteurization processes applied to human milk prior to delivery to neonates could affect the concentration of immunomodulatory proteins, especially lactoferrin, secretory immunoglobulin A, and lysozyme. Leptin was unaffected by the various handling processes tested. Fresh milk was found to be the best food for neonates. Further studies are warranted to evaluate the functional activity of these proteins and their effects on infants' immunological status.

  3. Kinetics of milk lipid droplet transport, growth, and secretion revealed by intravital imaging: lipid droplet release is intermittently stimulated by oxytocin | Center for Cancer Research

    Science.gov (United States)

    Description of the cover: The micrograph shows lipid droplets (red) accumulating at the apical surface of secretory cells (green) between oxytocin-induced contractions in a transgenic mouse line that expresses green fluorescent protein in the cytoplasm of most cells.

  4. Droplets, Bubbles and Ultrasound Interactions.

    Science.gov (United States)

    Shpak, Oleksandr; Verweij, Martin; de Jong, Nico; Versluis, Michel

    2016-01-01

    The interaction of droplets and bubbles with ultrasound has been studied extensively in the last 25 years. Microbubbles are broadly used in diagnostic and therapeutic medical applications, for instance, as ultrasound contrast agents. They have a similar size as red blood cells, and thus are able to circulate within blood vessels. Perfluorocarbon liquid droplets can be a potential new generation of microbubble agents as ultrasound can trigger their conversion into gas bubbles. Prior to activation, they are at least five times smaller in diameter than the resulting bubbles. Together with the violent nature of the phase-transition, the droplets can be used for local drug delivery, embolotherapy, HIFU enhancement and tumor imaging. Here we explain the basics of bubble dynamics, described by the Rayleigh-Plesset equation, bubble resonance frequency, damping and quality factor. We show the elegant calculation of the above characteristics for the case of small amplitude oscillations by linearizing the equations. The effect and importance of a bubble coating and effective surface tension are also discussed. We give the main characteristics of the power spectrum of bubble oscillations. Preceding bubble dynamics, ultrasound propagation is introduced. We explain the speed of sound, nonlinearity and attenuation terms. We examine bubble ultrasound scattering and how it depends on the wave-shape of the incident wave. Finally, we introduce droplet interaction with ultrasound. We elucidate the ultrasound-focusing concept within a droplets sphere, droplet shaking due to media compressibility and droplet phase-conversion dynamics.

  5. Electrohydrodynamic simulation of electrically controlled droplet generation

    International Nuclear Information System (INIS)

    Ouedraogo, Yun; Gjonaj, Erion; Weiland, Thomas; Gersem, Herbert De; Steinhausen, Christoph; Lamanna, Grazia; Weigand, Bernhard

    2017-01-01

    Highlights: • We develop a full electrohydrodynamic simulation approach which allows for the accurate modeling of droplet dynamics under the influence of transient electric fields. The model takes into account conductive, capacitive as well as convective electrical currents in the fluid. • Simulation results are shown for an electrically driven droplet generator using highly conductive acetone droplets and low conductivity pentane droplets, respectively. Excellent agreement with measurement is found. • We investigate the operation characteristic of the droplet generator by computing droplet sizes and detachment times with respect to the applied voltage. • The droplet charging effect is demonstrated for pentane droplets as well as for acetone droplets under long voltage pulses. We show that due to the very different relaxation times, the charging behavior of the two liquids is very different. • We demonstrate that due to this behavior, also the detachment mechanisms for acetone and pentane droplets are different. For low conductivity (pentane) droplets, droplet detachment is only possible after the electric fields are switched off. This is because the effective electric polarization force points upwards, thus, inhibiting the detachment of the droplet from the capillary tip. - Abstract: An electrohydrodynamic model for the simulation of droplet formation, detachment and motion in an electrically driven droplet generator is introduced. The numerical approach is based on the coupled solution of the multiphase flow problem with the charge continuity equation. For the latter, a modified convection-conduction model is applied, taking into account conductive, capacitive as well as convective electrical currents in the fluid. This allows for a proper description of charge relaxation phenomena in the moving fluid. In particular, the charge received by the droplet after detachment is an important parameter influencing the droplet dynamics in the test chamber

  6. High-resolution liquid patterns via three-dimensional droplet shape control.

    Science.gov (United States)

    Raj, Rishi; Adera, Solomon; Enright, Ryan; Wang, Evelyn N

    2014-09-25

    Understanding liquid dynamics on surfaces can provide insight into nature's design and enable fine manipulation capability in biological, manufacturing, microfluidic and thermal management applications. Of particular interest is the ability to control the shape of the droplet contact area on the surface, which is typically circular on a smooth homogeneous surface. Here, we show the ability to tailor various droplet contact area shapes ranging from squares, rectangles, hexagons, octagons, to dodecagons via the design of the structure or chemical heterogeneity on the surface. We simultaneously obtain the necessary physical insights to develop a universal model for the three-dimensional droplet shape by characterizing the droplet side and top profiles. Furthermore, arrays of droplets with controlled shapes and high spatial resolution can be achieved using this approach. This liquid-based patterning strategy promises low-cost fabrication of integrated circuits, conductive patterns and bio-microarrays for high-density information storage and miniaturized biochips and biosensors, among others.

  7. Biological activities and applications of dioscorins, the major tuber storage proteins of yam.

    Science.gov (United States)

    Lu, Yeh-Lin; Chia, Cho-Yun; Liu, Yen-Wenn; Hou, Wen-Chi

    2012-01-01

    Yam tubers, a common tuber crop and an important traditional Chinese medicine in Taiwan, have many bioactive substances, including phenolic compounds, mucilage polysaccharides, steroidal saponins and proteins. Among the total soluble proteins, 80% of them are dioscorins. In the past two decades, many studies showed that dioscorins exhibited biological activities both in vitro and in vivo, including the enzymatic, antioxidant, antihypertensive, immunomodulatory, lectin activities and the protecting role on airway epithelial cells against allergens in vitro. Some of these activities are survived after chemical, heating process or enzymatic digestion. Despite of lacking the intact structural information and the detail action mechanisms in the cells, yam dioscorins are potential resources for developing as functional foods and interesting targets for food protein researchers.

  8. Biological Activities and Applications of Dioscorins, the Major Tuber Storage Proteins of Yam

    Directory of Open Access Journals (Sweden)

    Yeh-Lin Lu

    2012-01-01

    Full Text Available Yam tubers, a common tuber crop and an important traditional Chinese medicine in Taiwan, have many bioactive substances, including phenolic compounds, mucilage polysaccharides, steroidal saponins and proteins. Among the total soluble proteins, 80% of them are dioscorins. In the past two decades, many studies showed that dioscorins exhibited biological activities both in vitro and in vivo, including the enzymatic, antioxidant, antihypertensive, immunomodulatory, lectin activities and the protecting role on airway epithelial cells against allergens in vitro. Some of these activities are survived after chemical, heating process or enzymatic digestion. Despite of lacking the intact structural information and the detail action mechanisms in the cells, yam dioscorins are potential resources for developing as functional foods and interesting targets for food protein researchers.

  9. CONTAMINATION OF ANTI-VEGF DRUGS FOR INTRAVITREAL INJECTION: How Do Repackaging and Newly Developed Syringes Affect the Amount of Silicone Oil Droplets and Protein Aggregates?

    Science.gov (United States)

    Schargus, Marc; Werner, Benjamin P; Geerling, Gerd; Winter, Gerhard

    2017-08-21

    The particle counts and the nature of particles of three different antivascular endothelial growth factor agents (VEGF) in different containers in a laboratory setting were compared. Original prefilled ranibizumab glass syringes, original vials with aflibercept, and repacked ready-to-use plastic syringes with bevacizumab from a compounding pharmacy and a compounding company (CC) were analyzed. Particle counts and size distributions were quantified by different particle characterization methods (nephelometry, light obscuration, Micro-Flow Imaging, nanotracking analysis, resonant mass measurement). Using high-performance size-exclusion chromatography (HP-SEC), levels of protein drug monomer and soluble aggregates were determined. Nearly all samples showed similar product quality. Light obscuration and Micro-Flow Imaging showed a 4-fold to 9-fold higher total particle count in compounding company bevacizumab (other samples up to 42,000 particles/mL). Nanotracking analysis revealed highest values for compounding company bevacizumab (6,375 million particles/mL). All containers showed similar amounts of silicone oil microdroplets. Ranibizumab showed lowest particle count of all tested agents with only one monomer peak in HP-SEC. Repackaged bevacizumab from different suppliers showed varying product quality. All three tested agents are available in similar quality regarding particulate purity and silicone oil microdroplet count. Repackaging can have a major impact on the quality.

  10. Storage influence on the functional, sensory and keeping quality of quality protein maize flour.

    Science.gov (United States)

    Shobha, D; Kumar, H V Dileep; Sreeramasetty, T A; Puttaramanaik; Gowda, K T Pandurange; Shivakumar, G B

    2014-11-01

    Apart from nutritional values functional and sensory properties affect the behavior of food system and its acceptability for consumption during storage. Hence keeping quality of maize flour (HQPM-7) with and without lime treatment(control) was studied in terms of functional (bulk density, pH, swelling capacity, water and oil absorption capacity, least gelation concentration, peroxide value), sensory (appearance, color, taste, texture, mouth feel and overall acceptability) and rolling parameters (water absorption by flour, rolling quality, diameter after baking ) for a period of 6 months under room temperature (25 ± 5 °C) in two types of packages viz, LDPE cover (P) and plastic box (B). Physical parameters such as length, breadth and thickness (11.26-10.52 mm, 9.67-9.14 mm, & 4.72-3.95 mm) were reduced in lime treated grains compared to control. Significant increase (p ≤ 0.05) in ash content of lime treated flour (1.67 ± 0.01 g) was observed compared to control (1.5 ± 0.02 g). Calcium content of lime treated maize flour increased significantly (p ≤ 0.05) from 48 to 136 mg. There is a significant reduction in functional properties of flour after 3 and 2 months irrespective in polyethylene cover and plastic box. The properties like rolling quality, diameter after baking and water uptake by the flour were reduced significantly (p ≤ 0.05) after 4 months of storage in treated and after 1 month in control samples. Sensory scores of roti (dry pan cake) decreased significantly after 3 months of storage with an overall acceptability score of 4.0 and 3.4. In control samples mean taste (3.6), mouth feel (3.8) as well as OAA scores (3.8) decreased after second month. Hence lime treated maize flour with added nutritional benefits is suitable for making rotis of good palatability and can be stored in LDPE covers up to 3 months.

  11. Increased lipid droplet accumulation associated with a peripheral sensory neuropathy.

    Science.gov (United States)

    Marshall, Lee L; Stimpson, Scott E; Hyland, Ryan; Coorssen, Jens R; Myers, Simon J

    2014-04-01

    Hereditary sensory neuropathy type 1 (HSN-1) is an autosomal dominant neurodegenerative disease caused by missense mutations in the SPTLC1 gene. The SPTLC1 protein is part of the SPT enzyme which is a ubiquitously expressed, critical and thus highly regulated endoplasmic reticulum bound membrane enzyme that maintains sphingolipid concentrations and thus contributes to lipid metabolism, signalling, and membrane structural functions. Lipid droplets are dynamic organelles containing sphingolipids and membrane bound proteins surrounding a core of neutral lipids, and thus mediate the intracellular transport of these specific molecules. Current literature suggests that there are increased numbers of lipid droplets and alterations of lipid metabolism in a variety of other autosomal dominant neurodegenerative diseases, including Alzheimer's and Parkinson's disease. This study establishes for the first time, a significant increase in the presence of lipid droplets in HSN-1 patient-derived lymphoblasts, indicating a potential connection between lipid droplets and the pathomechanism of HSN-1. However, the expression of adipophilin (ADFP), which has been implicated in the regulation of lipid metabolism, was not altered in lipid droplets from the HSN-1 patient-derived lymphoblasts. This appears to be the first report of increased lipid body accumulation in a peripheral neuropathy, suggesting a fundamental molecular linkage between a number of neurodegenerative diseases.

  12. The Use of Legume Seed for Expression and Storage of High Value Proteins

    Czech Academy of Sciences Publication Activity Database

    Moravec, Tomáš; Čeřovská, Noemi

    2014-01-01

    Roč. 50, č. 2 (2014), s. 69-76 ISSN 1212-1975 R&D Projects: GA ČR(CZ) GAP501/12/1761 Institutional support: RVO:61389030 Keywords : pea * pharming * protein bodies Subject RIV: EE - Microbiology, Virology Impact factor: 0.364, year: 2014 http://agriculturejournals.cz/publicFiles/124126.pdf

  13. Changes in microbial populations of WPC34 and WPC80 whey protein during long term storage

    Science.gov (United States)

    The use of whey protein (WPC34 and WPC80) as a food ingredient and as a base for making biodegradable products is increasing. The need to alleviate world hunger in arid and semi-arid regions demands that we investigate the behavior of native bacteria in these products, especially during long term st...

  14. Oleoplaning droplets on lubricated surfaces

    Science.gov (United States)

    Daniel, Dan; Timonen, Jaakko V. I.; Li, Ruoping; Velling, Seneca J.; Aizenberg, Joanna

    2017-10-01

    Recently, there has been much interest in using lubricated surfaces to achieve extreme liquid repellency: a foreign droplet immiscible with the underlying lubricant layer was shown to slide off at a small tilt angle behaviour was hypothesized to arise from a thin lubricant overlayer film sandwiched between the droplet and solid substrate, but this has not been observed experimentally. Here, using thin-film interference, we are able to visualize the intercalated film under both static and dynamic conditions. We further demonstrate that for a moving droplet, the film thickness follows the Landau-Levich-Derjaguin law. The droplet is therefore oleoplaning--akin to tyres hydroplaning on a wet road--with minimal dissipative force and no contact line pinning. The techniques and insights presented in this study will inform future work on the fundamentals of wetting for lubricated surfaces and enable their rational design.

  15. Droplet Translation Actuated by Photoelectrowetting.

    Science.gov (United States)

    Palma, Cesar; Deegan, Robert D

    2018-03-13

    In traditional electrowetting-on-dielectric (EWOD) devices, droplets are moved about a substrate using electric fields produced by an array of discrete electrodes. In this study, we show that a drop can be driven across a substrate with a localized light beam by exploiting the photoelectrowetting (PEW) effect, a light-activated variant of EWOD. Droplet transport actuated by PEW eliminates the need for electrode arrays and the complexities entailed in their fabrication and control, and offers a new approach for designing lab-on-a-chip applications. We report measurements of the maximum droplet speed as a function of frequency and magnitude of the applied bias, intensity of illumination, volume of the droplet, and viscosity and also introduce a model that reproduces these data.

  16. Soybean protein-based microparticles for oral delivery of probiotics with improved stability during storage and gut resistance.

    Science.gov (United States)

    González-Ferrero, C; Irache, J M; González-Navarro, C J

    2018-01-15

    The present work describes the encapsulation of probiotics using a by-product as wall material and a process feasible to be scaled-up: coacervation of soybean protein concentrate (SPC) by using calcium salts and spray-drying. SPC was extracted from soybean flour, produced during the processing of soybean milk, by alkaline extraction following isoelectric precipitation. Two probiotic strains were selected for encapsulation (Lactobacillus plantarum CECT 220 and Lactobacillus casei CECT 475) in order to evaluate the ability of SPC to encapsulate and protect bacteria from stress conditions. The viability of these encapsulated strains under in vitro gastrointestinal conditions and shelf-life during storage were compared with the most common forms commercialized nowadays. Results show that SPC is a feasible material for the development of probiotic microparticles with adequate physicochemical properties and enhanced significantly both probiotic viability and tolerance against simulated gastrointestinal fluids when compared to current available commercial forms. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. Quantum Nanostructures by Droplet Epitaxy

    OpenAIRE

    Somsak Panyakeow

    2009-01-01

    Droplet epitaxy is an alternative growth technique for several quantum nanostructures. Indium droplets are distributed randomly on GaAs substrates at low temperatures (120-350'C). Under background pressure of group V elements, Arsenic and Phosphorous, InAs and InP nanostructures are created. Quantum rings with isotropic shape are obtained at low temperature range. When the growth thickness is increased, quantum rings are transformed to quantum dot rings. At high temperature range, anisotropic...

  18. Lxr-driven enterocyte lipid droplet formation delays transport of ingested lipids.

    Science.gov (United States)

    Cruz-Garcia, Lourdes; Schlegel, Amnon

    2014-09-01

    Liver X receptors (Lxrs) are master regulators of cholesterol catabolism, driving the elimination of cholesterol from the periphery to the lumen of the intestine. Development of pharmacological agents to activate Lxrs has been hindered by synthetic Lxr agonists' induction of hepatic lipogenesis and hypertriglyceridemia. Elucidating the function of Lxrs in regulating enterocyte lipid handling might identify novel aspects of lipid metabolism that are pharmacologically amenable. We took a genetic approach centered on the single Lxr gene nr1h3 in zebrafish to study the role of Lxr in enterocyte lipid metabolism. Loss of nr1h3 function causes anticipated gene regulatory changes and cholesterol intolerance, collectively reflecting high evolutionary conservation of zebrafish Lxra function. Intestinal nr1h3 activation delays transport of absorbed neutral lipids, with accumulation of neutral lipids in enterocyte cytoplasmic droplets. This delay in transport of ingested neutral lipids protects animals from hypercholesterolemia and hepatic steatosis induced by a high-fat diet. On a gene regulatory level, Lxra induces expression of acsl3a, which encodes acyl-CoA synthetase long-chain family member 3a, a lipid droplet-anchored protein that directs fatty acyl chains into lipids. Forced overexpression of acls3a in enterocytes delays, in part, the appearance of neutral lipids in the vasculature of zebrafish larvae. Activation of Lxr in the intestine cell-autonomously regulates the rate of delivery of absorbed lipids by inducting a temporary lipid intestinal droplet storage depot. Copyright © 2014 by the American Society for Biochemistry and Molecular Biology, Inc.

  19. Dimers of G-Protein Coupled Receptors as Versatile Storage and Response Units

    Directory of Open Access Journals (Sweden)

    Michael S. Parker

    2014-03-01

    Full Text Available The status and use of transmembrane, extracellular and intracellular domains in oligomerization of heptahelical G-protein coupled receptors (GPCRs are reviewed and for transmembrane assemblies also supplemented by new experimental evidence. The transmembrane-linked GPCR oligomers typically have as the minimal unit an asymmetric ~180 kDa pentamer consisting of receptor homodimer or heterodimer and a G-protein αβγ subunit heterotrimer. With neuropeptide Y (NPY receptors, this assembly is converted to ~90 kDa receptor monomer-Gα complex by receptor and Gα agonists, and dimers/heteropentamers are depleted by neutralization of Gαi subunits by pertussis toxin. Employing gradient centrifugation, quantification and other characterization of GPCR dimers at the level of physically isolated and identified heteropentamers is feasible with labeled agonists that do not dissociate upon solubilization. This is demonstrated with three neuropeptide Y (NPY receptors and could apply to many receptors that use large peptidic agonists.

  20. Purification and partial characterization of storage proteins in Lupinus angustifolius seeds

    Directory of Open Access Journals (Sweden)

    Millán, Francisco

    2004-12-01

    Full Text Available Lupinus angustifolius seed proteins have been purified by sequential dialysis and ion exchange chromatography, and their amino acid composition has been studied in order to determine their nutritional value as sources of essential amino acids. Albumins include a great variety of proteins. Globulins were resolved in α, β and δ conglutins. Conglutin α is the main protein in the seeds of L. angustifolius, representing 76.6% of the total. While lysine was found to be the limiting amino acid in L. Angustifolius seed proteins as a whole, tyrosine was the limiting amino acid in albumins, and methione and lysine were limiting in globulins. Lysine, methionine and histidine were limiting amino acids in  α conglutin.Se ha realizado una purificación por diálisis secuencial y cromatografía de intercambio iónico de las proteínas de la semilla de L. angustifolius, y su composición en aminoácidos ha sido estudiada para determinar su valor nutricional como fuentes de aminoácidos esenciales. Las albúminas están compuestas por una compleja mezcla de proteínas. Las globulinas se resolvieron en conglutinas α, β y δ. La conglutina  α es la proteína más abundante en las semillas de L. angustifolius representando el 76.6% del total. Las proteínas de la semilla de L. angustifolius son limitantes en lisina. Las albúminas son limitantes en tirosina y las globulinas en metionina y lisina. La α conglutina es limitante en lisina, metionina e histidina.

  1. Preparation of Barley Storage Protein, Hordein, for Analytical Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis

    DEFF Research Database (Denmark)

    Doll, Hans; Andersen, Bente

    1981-01-01

    The extraction, reduction, and alkylation of barley hordein for routine electrophoresis in sodium dodecyl sulfate-polyacrylamide gels were studied to set up a simple preparation procedure giving well-resolved bands in the electrophoresis gel. Hordein was extracted from single crushed seeds or flour...... by aqueous 50% propan-2-ol containing a Tris-borate buffer, pH 8.6. The presence of the buffer facilitates the consecutive complete reduction of the extracted protein in the alcohol. Reduction and alkylation in the buffer containing propan-2-ol give sharper bands in the electrophoresis than reduction...

  2. Assessment of genetic diversity among some accessions of sage (Salvia officinalis L. using electrophoresis of seed storage proteins

    Directory of Open Access Journals (Sweden)

    Seyed Abbas Mirjalili

    2016-06-01

    Full Text Available The genus Salvia (Lamiaceae comprises over 900 species in the world, with a relatively wide dispersion in the Iran’s flora. Until now, about 58 species of the genus have been reported and identified in Iran, in which 17 of them were endemic. In order to study, investigate and evaluate the intraspecific diversity, similarity and dissimilarity among Iranian Salvia officinalis accessions, an experiment was carried out using SDS-PAGE technique. In this study, the seeds from five accessions were collected from gene bank and were evaluated. The seed storage proteins were extracted by buffers and were measured. Phylogenetic relationships were analyzed according to presence and absence of bands on the gel. A dendrogram was prepared using calculation of the accession’s similarity index. Mean comparison were done by Tukey’s test. The seed protein contents showed significant differences (p≤0.01 among accessions. A total of 39 bands were indicated on the gel. The maximum diversity was detected in the accession No. 2 while, the lowest band’s number were recorded with the accessions No. 1 and 5. Based on dendrogram, the accessions were divided into two groups; one includes accession No. 1 and 4 other accessions were located in the second group further classified into two subgroup including accessions No. 2 and 3 in one clade and accessions No. 4 and 5 in the other ones.

  3. Uniform-droplet spray forming

    Energy Technology Data Exchange (ETDEWEB)

    Blue, C.A.; Sikka, V.K. [Oak Ridge National Lab., TN (United States); Chun, Jung-Hoon [Massachusetts Institute of Technology, Cambridge, MA (United States); Ando, T. [Tufts Univ., Medford, MA (United States)

    1997-04-01

    The uniform-droplet process is a new method of liquid-metal atomization that results in single droplets that can be used to produce mono-size powders or sprayed-on to substrates to produce near-net shapes with tailored microstructure. The mono-sized powder-production capability of the uniform-droplet process also has the potential of permitting engineered powder blends to produce components of controlled porosity. Metal and alloy powders are commercially produced by at least three different methods: gas atomization, water atomization, and rotating disk. All three methods produce powders of a broad range in size with a very small yield of fine powders with single-sized droplets that can be used to produce mono-size powders or sprayed-on substrates to produce near-net shapes with tailored microstructures. The economical analysis has shown the process to have the potential of reducing capital cost by 50% and operating cost by 37.5% when applied to powder making. For the spray-forming process, a 25% savings is expected in both the capital and operating costs. The project is jointly carried out at Massachusetts Institute of Technology (MIT), Tuffs University, and Oak Ridge National Laboratory (ORNL). Preliminary interactions with both finished parts and powder producers have shown a strong interest in the uniform-droplet process. Systematic studies are being conducted to optimize the process parameters, understand the solidification of droplets and spray deposits, and develop a uniform-droplet-system (UDS) apparatus appropriate for processing engineering alloys.

  4. Accelerating Yeast Prion Biology using Droplet Microfluidics

    Science.gov (United States)

    Ung, Lloyd; Rotem, Assaf; Jarosz, Daniel; Datta, Manoshi; Lindquist, Susan; Weitz, David

    2012-02-01

    Prions are infectious proteins in a misfolded form, that can induce normal proteins to take the misfolded state. Yeast prions are relevant, as a model of human prion diseases, and interesting from an evolutionary standpoint. Prions may also be a form of epigenetic inheritance, which allow yeast to adapt to stressful conditions at rates exceeding those of random mutations and propagate that adaptation to their offspring. Encapsulation of yeast in droplet microfluidic devices enables high-throughput measurements with single cell resolution, which would not be feasible using bulk methods. Millions of populations of yeast can be screened to obtain reliable measurements of prion induction and loss rates. The population dynamics of clonal yeast, when a fraction of the cells are prion expressing, can be elucidated. Furthermore, the mechanism by which certain strains of bacteria induce yeast to express prions in the wild can be deduced. Integrating the disparate fields of prion biology and droplet microfluidics reveals a more complete picture of how prions may be more than just diseases and play a functional role in yeast.

  5. Self-assembly of caseinomacropeptide as a potential key mechanism in the formation of visible storage induced aggregates in acidic whey protein isolate dispersions

    DEFF Research Database (Denmark)

    Villumsen, Nanna Stengaard; Jensen, Hanne Bak; Thu Le, Thao Thi

    2015-01-01

    Visible aggregates formed during storage in acidic whey protein isolate (WPI) dispersions represent a challenge to the beverage industry. Batch-to-batch variations are observed that prevents consistent quality and shelf-life prediction. Heat-treatment of WPI dispersions at 120°C for 20s instead...

  6. Effect of storage temperature on survival and recovery of thermal and extrusion injured Escherichia coli populations in whey protein concentrate and corn meal

    Science.gov (United States)

    In a previous study, we reported viability loss of Escherichia coli populations in corn (CP) and whey protein products (WPP) extruded at different temperatures. However, information on the effect of storage temperatures on injured bacterial populations was not addressed. The objective of this study ...

  7. Effects of heat stress and probiotic supplementation on protein functionality and oxidative stability of ground chicken leg meat during display storage

    Science.gov (United States)

    The objective of this study was to evaluate effects of heat stress and probiotic supplementation on protein functionality and oxidative stability of ground chicken leg meat during display storage. Two hundred and forty 1-day-old male chicks (5 bird per pen) were randomly subjected to four treatments...

  8. A Droplet Microfluidic Platform for Automating Genetic Engineering.

    Science.gov (United States)

    Gach, Philip C; Shih, Steve C C; Sustarich, Jess; Keasling, Jay D; Hillson, Nathan J; Adams, Paul D; Singh, Anup K

    2016-05-20

    We present a water-in-oil droplet microfluidic platform for transformation, culture and expression of recombinant proteins in multiple host organisms including bacteria, yeast and fungi. The platform consists of a hybrid digital microfluidic/channel-based droplet chip with integrated temperature control to allow complete automation and integration of plasmid addition, heat-shock transformation, addition of selection medium, culture, and protein expression. The microfluidic format permitted significant reduction in consumption (100-fold) of expensive reagents such as DNA and enzymes compared to the benchtop method. The chip contains a channel to continuously replenish oil to the culture chamber to provide a fresh supply of oxygen to the cells for long-term (∼5 days) cell culture. The flow channel also replenished oil lost to evaporation and increased the number of droplets that could be processed and cultured. The platform was validated by transforming several plasmids into Escherichia coli including plasmids containing genes for fluorescent proteins GFP, BFP and RFP; plasmids with selectable markers for ampicillin or kanamycin resistance; and a Golden Gate DNA assembly reaction. We also demonstrate the applicability of this platform for transformation in widely used eukaryotic organisms such as Saccharomyces cerevisiae and Aspergillus niger. Duration and temperatures of the microfluidic heat-shock procedures were optimized to yield transformation efficiencies comparable to those obtained by benchtop methods with a throughput up to 6 droplets/min. The proposed platform offers potential for automation of molecular biology experiments significantly reducing cost, time and variability while improving throughput.

  9. Potential costs of bacterial infection on storage protein gene expression and reproduction in queenless Apis mellifera worker bees on distinct dietary regimes.

    Science.gov (United States)

    Lourenço, Anete Pedro; Martins, Juliana Ramos; Guidugli-Lazzarini, Karina Rosa; Macedo, Liliane Maria Fróes; Bitondi, Márcia Maria Gentile; Simões, Zilá Luz Paulino

    2012-09-01

    Insects are able to combat infection by initiating an efficient immune response that involves synthesizing antimicrobial peptides and a range of other defense molecules. These responses may be costly to the organism, resulting in it exploiting endogenous resources to maintain homeostasis or support defense to the detriment of other physiological needs. We used queenless worker bees on distinct dietary regimes that may alter hemolymph protein storage and ovary activation to investigate the physiological costs of infection with Serratia marcescens. The expression of the genes encoding the storage proteins vitellogenin and hexamerin 70a, the vitellogenin receptor, and vasa (which has a putative role in reproduction), was impaired in the infected bees. This impairment was mainly evident in the bees fed beebread, which caused significantly higher expression of these genes than did royal jelly or syrup, and this was confirmed at the vitellogenin and hexamerin 70a protein levels. Beebread was also the only diet that promoted ovary activation in the queenless bees, but this activation was significantly impaired by the infection. The expression of the genes encoding the storage proteins apolipophorins-I and -III and the lipophorin receptor was not altered by infection regardless the diet provided to the bees. Similarly, the storage of apolipophorin-I in the hemolymph was only slightly impaired by the infection, independently of the supplied diet. Taken together these results indicate that, infection demands a physiological cost from the transcription of specific protein storage-related genes and from the reproductive capacity. Copyright © 2012 Elsevier Ltd. All rights reserved.

  10. Instability of expanding bacterial droplets.

    Science.gov (United States)

    Sokolov, Andrey; Rubio, Leonardo Dominguez; Brady, John F; Aranson, Igor S

    2018-04-03

    Suspensions of motile bacteria or synthetic microswimmers, termed active matter, manifest a remarkable propensity for self-organization, and formation of large-scale coherent structures. Most active matter research deals with almost homogeneous in space systems and little is known about the dynamics of strongly heterogeneous active matter. Here we report on experimental and theoretical studies on the expansion of highly concentrated bacterial droplets into an ambient bacteria-free fluid. The droplet is formed beneath a rapidly rotating solid macroscopic particle inserted in the suspension. We observe vigorous instability of the droplet reminiscent of a violent explosion. The phenomenon is explained in terms of continuum first-principle theory based on the swim pressure concept. Our findings provide insights into the dynamics of active matter with strong density gradients and significantly expand the scope of experimental and analytic tools for control and manipulation of active systems.

  11. An Overview of Lipid Droplets in Cancer and Cancer Stem Cells

    KAUST Repository

    Tirinato, Luca; Pagliari, Francesca; Limongi, Tania; Marini, Monica; Falqui, Andrea; Seco, J.; Candeloro, Patrizio; Liberale, Carlo; Di Fabrizio, Enzo M.

    2017-01-01

    For decades, lipid droplets have been considered as the main cellular organelles involved in the fat storage, because of their lipid composition. However, in recent years, some new and totally unexpected roles have been discovered for them: (i) they are active sites for synthesis and storage of inflammatory mediators, and (ii) they are key players in cancer cells and tissues, especially in cancer stem cells. In this review, we summarize the main concepts related to the lipid droplet structure and function and their involvement in inflammatory and cancer processes.

  12. An Overview of Lipid Droplets in Cancer and Cancer Stem Cells

    KAUST Repository

    Tirinato, Luca

    2017-08-13

    For decades, lipid droplets have been considered as the main cellular organelles involved in the fat storage, because of their lipid composition. However, in recent years, some new and totally unexpected roles have been discovered for them: (i) they are active sites for synthesis and storage of inflammatory mediators, and (ii) they are key players in cancer cells and tissues, especially in cancer stem cells. In this review, we summarize the main concepts related to the lipid droplet structure and function and their involvement in inflammatory and cancer processes.

  13. The effect of melanin-free extract from Sepia esculenta ink on lipid peroxidation, protein oxidation and water-holding capacity of tilapia fillet during cold storage.

    Science.gov (United States)

    Duan, Zhen-Hua; Liu, Hua-Zhong; Luo, Ping; Gu, Yi-Peng; Li, Yan-Qun

    2018-03-14

    Preservative effect of melanin-free extract of Sepia esculenta ink (MFESI) on Sparus latus fillet has been verified in our previous work. This study aims to further approach the mechanism of MFESI for extending the shelf-life of fish fillet during cold storage. Tilapia fillets were treated with different dosage of MFESI (0, 15, 25 and 35 mg/ml) and packed with preservative film for succedent cold-storage at 4 °C for scheduled time. Contents of total volatile basic nitrogen and sulfydryl and carbanyl groups were measured for evaluating protein oxidation. Malondialdehyde contents were measured for estimating lipid peroxidation and loss of water was used to determine water-holding capacity of fillet. The data indicated that MFESI not only possessed certain degree of antioxidant capacity in vitro, also lengthened shelf-life of tilapia fillet in cold-storage condition. Apart from 15 mg/ml, both 25 and 35 mg/ml of MFESI obviously prevented lipid and protein from oxidation and reduced loss of water from tilapia fillets, and the latter was more effective than the former. MFESI can repress lipid peroxidation and protein oxidation and reduce water loss, maintain the tilapia fillets quality and, thus, it could be an effective and natural preservative for extending the shelf-life of tilapia fillets during cold storage.

  14. Dual-nozzle microfluidic droplet generator

    Science.gov (United States)

    Choi, Ji Wook; Lee, Jong Min; Kim, Tae Hyun; Ha, Jang Ho; Ahrberg, Christian D.; Chung, Bong Geun

    2018-05-01

    The droplet-generating microfluidics has become an important technique for a variety of applications ranging from single cell analysis to nanoparticle synthesis. Although there are a large number of methods for generating and experimenting with droplets on microfluidic devices, the dispensing of droplets from these microfluidic devices is a challenge due to aggregation and merging of droplets at the interface of microfluidic devices. Here, we present a microfluidic dual-nozzle device for the generation and dispensing of uniform-sized droplets. The first nozzle of the microfluidic device is used for the generation of the droplets, while the second nozzle can accelerate the droplets and increase the spacing between them, allowing for facile dispensing of droplets. Computational fluid dynamic simulations were conducted to optimize the design parameters of the microfluidic device.

  15. Taking advantage of reduced droplet-surface interaction to optimize transport of bioanalytes in digital microfluidics.

    Science.gov (United States)

    Freire, Sergio L S; Thorne, Nathaniel; Wutkowski, Michael; Dao, Selina

    2014-11-10

    Digital microfluidics (DMF), a technique for manipulation of droplets, is a promising alternative for the development of "lab-on-a-chip" platforms. Often, droplet motion relies on the wetting of a surface, directly associated with the application of an electric field; surface interactions, however, make motion dependent on droplet contents, limiting the breadth of applications of the technique. Some alternatives have been presented to minimize this dependence. However, they rely on the addition of extra chemical species to the droplet or its surroundings, which could potentially interact with droplet moieties. Addressing this challenge, our group recently developed Field-DW devices to allow the transport of cells and proteins in DMF, without extra additives. Here, the protocol for device fabrication and operation is provided, including the electronic interface for motion control. We also continue the studies with the devices, showing that multicellular, relatively large, model organisms can also be transported, arguably unaffected by the electric fields required for device operation.

  16. Colliding droplets: A short film presentation

    Science.gov (United States)

    Hendricks, C. D.

    1981-12-01

    A series of experiments were performed in which liquid droplets were caused to collide. Impact velocities to several meters per second and droplet diameters up to 600 micrometers were used. The impact parameters in the collisions vary from zero to greater than the sum of the droplet radii. Photographs of the collisions were taken with a high speed framing camera in order to study the impacts and subsequent behavior of the droplets.

  17. Numerical study of droplet evaporation in an acoustic levitator

    Science.gov (United States)

    Bänsch, Eberhard; Götz, Michael

    2018-03-01

    We present a finite element method for the simulation of all relevant processes of the evaporation of a liquid droplet suspended in an acoustic levitation device. The mathematical model and the numerical implementation take into account heat and mass transfer across the interface between the liquid and gaseous phase and the influence of acoustic streaming on this process, as well as the displacement and deformation of the droplet due to acoustic radiation pressure. We apply this numerical method to several theoretical and experimental examples and compare our results with the well-known d2-law for the evaporation of spherical droplets and with theoretical predictions for the acoustic streaming velocity. We study the influence of acoustic streaming on the distribution of water vapor and temperature in the levitation device, with special attention to the vapor distribution in the emerging toroidal vortices. We also compare the evaporation rate of a droplet with and without acoustic streaming, as well as the evaporation rates in dependence of different temperatures and sound pressure levels. Finally, a simple model of protein inactivation due to heat damage is considered and studied for different evaporation settings and their respective influence on protein damage.

  18. Droplet centrifugation, droplet DNA extraction, and rapid droplet thermocycling for simpler and faster PCR assay using wire-guided manipulations.

    Science.gov (United States)

    You, David J; Yoon, Jeong-Yeol

    2012-09-04

    A computer numerical control (CNC) apparatus was used to perform droplet centrifugation, droplet DNA extraction, and rapid droplet thermocycling on a single superhydrophobic surface and a multi-chambered PCB heater. Droplets were manipulated using "wire-guided" method (a pipette tip was used in this study). This methodology can be easily adapted to existing commercial robotic pipetting system, while demonstrated added capabilities such as vibrational mixing, high-speed centrifuging of droplets, simple DNA extraction utilizing the hydrophobicity difference between the tip and the superhydrophobic surface, and rapid thermocycling with a moving droplet, all with wire-guided droplet manipulations on a superhydrophobic surface and a multi-chambered PCB heater (i.e., not on a 96-well plate). Serial dilutions were demonstrated for diluting sample matrix. Centrifuging was demonstrated by rotating a 10 μL droplet at 2300 round per minute, concentrating E. coli by more than 3-fold within 3 min. DNA extraction was demonstrated from E. coli sample utilizing the disposable pipette tip to cleverly attract the extracted DNA from the droplet residing on a superhydrophobic surface, which took less than 10 min. Following extraction, the 1500 bp sequence of Peptidase D from E. coli was amplified using rapid droplet thermocycling, which took 10 min for 30 cycles. The total assay time was 23 min, including droplet centrifugation, droplet DNA extraction and rapid droplet thermocycling. Evaporation from of 10 μL droplets was not significant during these procedures, since the longest time exposure to air and the vibrations was less than 5 min (during DNA extraction). The results of these sequentially executed processes were analyzed using gel electrophoresis. Thus, this work demonstrates the adaptability of the system to replace many common laboratory tasks on a single platform (through re-programmability), in rapid succession (using droplets), and with a high level of

  19. Droplet centrifugation, droplet DNA extraction, and rapid droplet thermocycling for simpler and faster PCR assay using wire-guided manipulations

    Directory of Open Access Journals (Sweden)

    You David J

    2012-09-01

    Full Text Available Abstract A computer numerical control (CNC apparatus was used to perform droplet centrifugation, droplet DNA extraction, and rapid droplet thermocycling on a single superhydrophobic surface and a multi-chambered PCB heater. Droplets were manipulated using “wire-guided” method (a pipette tip was used in this study. This methodology can be easily adapted to existing commercial robotic pipetting system, while demonstrated added capabilities such as vibrational mixing, high-speed centrifuging of droplets, simple DNA extraction utilizing the hydrophobicity difference between the tip and the superhydrophobic surface, and rapid thermocycling with a moving droplet, all with wire-guided droplet manipulations on a superhydrophobic surface and a multi-chambered PCB heater (i.e., not on a 96-well plate. Serial dilutions were demonstrated for diluting sample matrix. Centrifuging was demonstrated by rotating a 10 μL droplet at 2300 round per minute, concentrating E. coli by more than 3-fold within 3 min. DNA extraction was demonstrated from E. coli sample utilizing the disposable pipette tip to cleverly attract the extracted DNA from the droplet residing on a superhydrophobic surface, which took less than 10 min. Following extraction, the 1500 bp sequence of Peptidase D from E. coli was amplified using rapid droplet thermocycling, which took 10 min for 30 cycles. The total assay time was 23 min, including droplet centrifugation, droplet DNA extraction and rapid droplet thermocycling. Evaporation from of 10 μL droplets was not significant during these procedures, since the longest time exposure to air and the vibrations was less than 5 min (during DNA extraction. The results of these sequentially executed processes were analyzed using gel electrophoresis. Thus, this work demonstrates the adaptability of the system to replace many common laboratory tasks on a single platform (through re-programmability, in rapid succession (using droplets

  20. Some Physics Inside Drying Droplets

    Indian Academy of Sciences (India)

    IAS Admin

    trial applications in food, biochemical or soil sciences. .... Take a metal spoon (or any other surface that does ..... samples. The shape of the suspended particles inside the droplet itself can be used to eliminate the coffee ring effect. For example ...

  1. Vapor-droplet flow equations

    International Nuclear Information System (INIS)

    Crowe, C.T.

    1975-01-01

    General features of a vapor-droplet flow are discussed and the equations expressing the conservation of mass, momentum, and energy for the vapor, liquid, and mixture using the control volume approach are derived. The phenomenological laws describing the exchange of mass, momentum, and energy between phases are also reviewed. The results have application to development of water-dominated geothermal resources

  2. Droplets, Bubbles and Ultrasound Interactions

    NARCIS (Netherlands)

    Shpak, O.; Verweij, M.; de Jong, N.; Versluis, Michel; Escoffre, J.M.; Bouakaz, A.

    2016-01-01

    The interaction of droplets and bubbles with ultrasound has been studied extensively in the last 25 years. Microbubbles are broadly used in diagnostic and therapeutic medical applications, for instance, as ultrasound contrast agents. They have a similar size as red blood cells, and thus are able to

  3. Liquid droplet radiator performance studies

    Science.gov (United States)

    Mattick, A. T.; Hertzberg, A.

    By making use of droplets rather than solid surfaces to radiate waste heat in space, the liquid droplet radiator (LDR) achieves a radiating area/mass much larger than that of conventional radiators which use fins or heat pipes. The lightweight potential of the LDR is shown to be limited primarily by the radiative properties of the droplets. The requirement that the LDR heat transfer fluid have a very low vapor pressure limits the choice of fluids to relatively few—several liquid metals and Dow 705 silicone fluid are the only suitable candidates so far identified. An experimental determination of the emittance of submillimeter droplets of Dow 705 fluid indicates than an LDR using this fluid at temperatures of 275-335 K would be ⋍ 10 times lighter than the lightest solid surface radiators. Although several liquid metals appear to offer excellent performance in LDR applications at temperatures between 200 K and 975 K, experimental determination of liquid metal emissivities is needed for a conclusive assessment.

  4. Droplet based cavities and lasers

    DEFF Research Database (Denmark)

    Mølhave, Kristian; Kristensen, Anders; Mortensen, Asger

    2009-01-01

    The self-organized and molecularly smooth surface on liquid microdroplets makes them attractive as optical cavities with very high quality factors. This chapter describes the basic theory of optical modes in spherical droplets. The mechanical properties including vibrational excitation are also d...

  5. Spin lattices of walking droplets

    Science.gov (United States)

    Saenz, Pedro; Pucci, Giuseppe; Goujon, Alexis; Dunkel, Jorn; Bush, John

    2017-11-01

    We present the results of an experimental investigation of the spontaneous emergence of collective behavior in spin lattice of droplets walking on a vibrating fluid bath. The bottom topography consists of relatively deep circular wells that encourage the walking droplets to follow circular trajectories centered at the lattice sites, in one direction or the other. Wave-mediated interactions between neighboring drops are enabled through a thin fluid layer between the wells. The sense of rotation of the walking droplets may thus become globally coupled. When the coupling is sufficiently strong, interactions with neighboring droplets may result in switches in spin that lead to preferred global arrangements, including correlated (all drops rotating in the same direction) or anti-correlated (neighboring drops rotating in opposite directions) states. Analogies with ferromagnetism and anti-ferromagnetism are drawn. Different spatial arrangements are presented in 1D and 2D lattices to illustrate the effects of topological frustration. This work was supported by the US National Science Foundation through Grants CMMI-1333242 and DMS-1614043.

  6. Some Physics Inside Drying Droplets

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education; Volume 19; Issue 2. Some Physics Inside Drying Droplets. Dileep Mampallil. General Article Volume 19 Issue 2 February 2014 pp 123-134. Fulltext. Click here to view fulltext PDF. Permanent link: https://www.ias.ac.in/article/fulltext/reso/019/02/0123-0134 ...

  7. Easy route to superhydrophobic copper-based wire-guided droplet microfluidic systems.

    Science.gov (United States)

    Mumm, Florian; van Helvoort, Antonius T J; Sikorski, Pawel

    2009-09-22

    Droplet-based microfluidic systems are an expansion of the lab on a chip concept toward flexible, reconfigurable setups based on the modification and analysis of individual droplets. Superhydrophobic surfaces are one suitable candidate for the realization of droplet-based microfluidic systems as the high mobility of aqueous liquids on such surfaces offers possibilities to use novel or more efficient approaches to droplet movement. Here, copper-based superhydrophobic surfaces were produced either by the etching of polycrystalline copper samples along the grain boundaries using etchants common in the microelectronics industry, by electrodeposition of copper films with subsequent nanowire decoration based on thermal oxidization, or by a combination of both. The surfaces could be easily hydrophobized with thiol-modified fluorocarbons, after which the produced surfaces showed a water contact angle as high as 171 degrees +/- 2 degrees . As copper was chosen as the base material, established patterning techniques adopted from printed circuit board fabrication could be used to fabricate macrostructures on the surfaces with the intention to confine the droplets and, thus, to reduce the system's sensitivity to tilting and vibrations. A simple droplet-based microfluidic chip with inlets, outlets, sample storage, and mixing areas was produced. Wire guidance, a relatively new actuation method applicable to aqueous liquids on superhydrophobic surfaces, was applied to move the droplets.

  8. Effects of diet, packaging, and irradiation on protein oxidation, lipid oxidation, and color of raw broiler thigh meat during refrigerated storage.

    Science.gov (United States)

    Xiao, S; Zhang, W G; Lee, E J; Ma, C W; Ahn, D U

    2011-06-01

    This study was designed to evaluate the effects of dietary treatment, packaging, and irradiation singly or in combination on the oxidative stability of broiler chicken thigh meat. A total of 120 four-week-old chickens were divided into 12 pens (10 birds/pen), and 4 pens of broilers were randomly assigned to a control oxidized diet (5% oxidized oil) or an antioxidant-added diet [500 IU of vitamin E + 200 mg/kg of butylated hydroxyanisole (BHA)] and fed for 2 wk. After slaughter, thigh meats were separated, ground, packaged in either oxygen-permeable or oxygen-impermeable vacuum bags, and irradiated at 0 or 3 kGy. Lipid oxidation (TBA-reactive substances), protein oxidation (carbonyl), and color of the meat were measured at 1, 4, and 7 d of refrigerated storage. The lipid and protein oxidation of thigh meats from birds fed the diet supplemented with antioxidants (vitamin E + BHA) was significantly lower than the lipid and protein oxidation of birds fed the control diet, whereas the lipid and protein oxidation of broilers fed the oxidized oil diet was higher than that of birds fed the control diet. Vacuum packaging slowed, but irradiation accelerated, the lipid and protein oxidation of thigh meat during storage. Dietary antioxidants (vitamin E + BHA) and irradiation treatments showed a stronger effect on lipid oxidation than on protein oxidation. A significant correlation between lipid and protein oxidation in meat was found during storage. Dietary supplementation of vitamin E + BHA and the irradiation treatment increased the lightness and redness of thigh meat, respectively. It is suggested that appropriate use of dietary antioxidants in combination with packaging could be effective in minimizing oxidative changes in irradiated raw chicken thigh meat.

  9. Acetonitrile as a buffer additive for free zone capillary electrophoresis separation and characterization of maize (Zeamays L. ) and sorghum (Sorghum bicolor L. Moench) storage proteins.

    Science.gov (United States)

    Bean, S R; Lookhart, G L; Bietz, J A

    2000-02-01

    An improved method for separating and characterizing maize (Zea mays L.) and sorghum (Sorghum bicolor L. Moench) storage proteins by free zone capillary electrophoresis (FZCE) was developed. Previous electrophoretic methods for analyzing these proteins required high concentrations of urea to maintain protein solubility during separation. To overcome disadvantages of urea, we developed a FZCE method that mimicked reversed-phase high-performance liquid chromatography (RP-HPLC) in that it used high levels of acetonitrile (ACN) at low pH. The optimized FZCE buffer system consisted of 80 mM phosphate-glycine buffer, nominal pH 2.5, containing 60% ACN and a cellulose derivative to dynamically coat capillary walls. Resolution was similar to or higher than that previously achieved by FZCE buffers utilizing 8 M urea as a buffer additive. ACN concentrations of at least 50% were necessary to achieve acceptable separations; this ACN concentration is approximately that necessary to extract these storage proteins. ACN was equally effective as traditional ethanol solvents and 8 M urea for solubilizing maize and sorghum proteins. The ACN-based FZCE buffer system gave high repeatability (buffers. This FZCE method may be applicable for the analysis of other hydrophobic proteins without the use of urea.

  10. Lepidopteran defence droplets - a composite physical and chemical weapon against potential predators

    DEFF Research Database (Denmark)

    Pentzold, Stefan; Zagrobelny, Mika; Khakimov, Bekzod

    2016-01-01

    predators together and immobilised them. Droplets were characterised by a matrix of an aqueous solution of glycine-rich peptides (H-WG11 -NH2) with significant amounts of proteins and glucose. Among the proteins, defensive proteins such as protease inhibitors, proteases and oxidases were abundant...

  11. Energy storage

    International Nuclear Information System (INIS)

    Anon.

    1992-01-01

    This chapter discusses the role that energy storage may have on the energy future of the US. The topics discussed in the chapter include historical aspects of energy storage, thermal energy storage including sensible heat storage, latent heat storage, thermochemical heat storage, and seasonal heat storage, electricity storage including batteries, pumped hydroelectric storage, compressed air energy storage, and superconducting magnetic energy storage, and production and combustion of hydrogen as an energy storage option

  12. Mice lacking lipid droplet-associated hydrolase, a gene linked to human prostate cancer, have normal cholesterol ester metabolism

    DEFF Research Database (Denmark)

    Kory, Nora; Grond, Susanne; Kamat, Siddhesh S

    2017-01-01

    Variations in the gene LDAH (C2ORF43), which encodes lipid droplet-associated hydrolase (LDAH), are among few loci associated with human prostate cancer. Homologs of LDAH have been identified as proteins of lipid droplets (LDs). LDs are cellular organelles that store neutral lipids...

  13. Adipophilin distribution and colocalization with lipid droplets in skeletal muscle.

    LENUS (Irish Health Repository)

    Shaw, Christopher S

    2009-05-01

    Intramyocellular lipids (IMCL) are stored as discrete lipid droplets which are associated with a number of proteins. The lipid droplet-associated protein adipophilin (the human orthologue of adipose differentiation-related protein) is ubiquitously expressed and is one of the predominant lipid droplet-proteins in skeletal muscle. The aim of this study was to investigate the subcellular distribution of adipophilin in human muscle fibres and to measure the colocalization of adipophilin with IMCL. Muscle biopsies from six lean male cyclists (BMI 23.4 +\\/- 0.4, aged 31 +\\/- 2 years, W (max) 346 +\\/- 8) were stained for myosin heavy chain type 1, IMCL, adipophilin and mitochondria using immunofluorescence and viewed with widefield and confocal fluorescence microscopy. The present study shows that like IMCL, the adipophilin content is ~twofold greater in type I skeletal muscle fibres and is situated in the areas between the mitochondrial network. Colocalization analysis demonstrated that 61 +\\/- 2% of IMCL contain adipophilin. Although the majority of adipophilin is contained within IMCL, 36 +\\/- 4% of adipophilin is not associated with IMCL. In conclusion, this study indicates that the IMCL pool is heterogeneous, as the majority but not all IMCL contain adipophilin.

  14. Effect of ice storage on muscle protein properties and qualities of emulsion fish sausage from bigeye snapper (Priacanthus tayenus and lizardfish (Saurida undosquamis

    Directory of Open Access Journals (Sweden)

    Vittayanont, M.

    2005-01-01

    Full Text Available The chemical changes in fish muscle and natural actomyosin (NAM from bigeye snapper (Priacanthus tayenus and lizardfish (Saurida undosquamis muscle during 0, 2, 4, 6, 8, 10, 12 and 14 days of iced storage were studied. Myosin heavy chain (MHC of NAM extracted from two fish species was degraded throughout iced storage. However, no changes in actin were observed. The total volatile base (TVB. trimethylamine (TMA and surface hydrophobicity increased, while the total sulfhydryl content and emulsion capacity of NAM from both fish species decreased significantly as the storage time increased (p<0.05. A Texture ProfileAnalysis (TPA and shear force of emulsion fish sausages prepared from two fish species kept in ice for 0, 4, 8 and 12 days were investigated. The results showed that hardness, cohesiveness, gumminess, chewiness and shear force of sausage prepared from fish kept in ice were lower than those produced from fresh fish. However, no significant differences in adhesiveness were observed. Cooking loss of emulsion fish sausage from two fish species increased throughout storage time (p<0.05. The texture of bigeye snapper sausage was better than that of lizardfish sausages. Scanning electron microscopy (SEM micrographs of emulsion fish sausage from two fish species revealed bigger voids, thicker strands and less continuity of protein strands with increasing storage time. More microstructural changes were observed in sausages from lizardfish, compared to those in sausages from bigeye snapper.

  15. Effect of harvest period on the quality of storage roots and protein content of the leaves in five cassava cultivars (Manihot esculenta, Crantz

    Directory of Open Access Journals (Sweden)

    Sagrilo Edvaldo

    2003-01-01

    Full Text Available The effect of harvest period on the quality of storage roots and leaves of cassava cultivars was determined in an experiment carried out in a randomized complete block design with four replications in a split plot scheme, with five cultivars in the plots and ten harvest times in the subplots. The IAC 13 cultivar had the highest rate of dry matter accumulation in the storage roots and the Mico cultivar the lowest. The period of least dry matter content in the storage roots occurred later for the Fécula Branca, Mico and IAC 14 cultivars, and the minimum starch content in the storage roots occurred later for the Fécula Branca and Mico cultivars. In general, the IAC 13, IAC 14 and Fécula Branca cultivars had higher dry matter content in the storage roots, while higher starch content in the dry and fresh matter were obtained in the Fécula Branca cultivar. The crude protein content in the leaves decreased as the plant aged.

  16. Dancing droplets: Contact angle, drag, and confinement

    Science.gov (United States)

    Benusiglio, Adrien; Cira, Nate; Prakash, Manu

    2015-11-01

    When deposited on a clean glass slide, a mixture of water and propylene glycol forms a droplet of given contact angle, when both pure liquids spread. (Cira, Benusiglio, Prakash: Nature, 2015). The droplet is stabilized by a gradient of surface tension due to evaporation that induces a Marangoni flow from the border to the apex of the droplets. The apparent contact angle of the droplets depends on both their composition and the external humidity as captured by simple models. These droplets present remarkable properties such as lack of a large pinning force. We discuss the drag on these droplets as a function of various parameters. We show theoretical and experimental results of how various confinement geometries change the vapor gradient and the dynamics of droplet attraction.

  17. The regulation of lipid droplet size and phospholipid composition by stearoyl-CoA desaturase

    DEFF Research Database (Denmark)

    Shi, Xun; Li, Juan; Zou, Xiaoju

    2013-01-01

    Fatty acid desaturation regulates membrane function and fat storage in animals. To determine the contribution of stearoyl-CoA desaturase (SCD) activity on fat storage and development in the nematode Caenorhabditis elegans, we analyzed the lipid composition and lipid droplet size in the fat-6;fat-7...... desaturase mutants, independently, and in combination with mutants disrupted in conserved lipid metabolic pathways. C. elegans with impaired SCD activity displayed both reduced fat stores and decreased lipid droplet size. Mutants in the daf-2 (insulin-like growth factor receptor), rsks-1 (homolog of p70S6......-2;fat-6;fat-7 triple mutants, which had increased de novo fatty acid synthesis and wild type levels of fat stores. Notably, stearoyl-CoA desaturase activity is required for the formation of large-sized lipid droplets in all mutant backgrounds, as well as for normal ratios of phosphatidylcholine (PC...

  18. The Phospholipid:Diacylglycerol Acyltransferase Lro1 Is Responsible for Hepatitis C Virus Core-Induced Lipid Droplet Formation in a Yeast Model System.

    Directory of Open Access Journals (Sweden)

    Shingo Iwasa

    Full Text Available Chronic infection with the hepatitis C virus frequently induces steatosis, which is a significant risk factor for liver pathogenesis. Steatosis is characterized by the accumulation of lipid droplets in hepatocytes. The structural protein core of the virus induces lipid droplet formation and localizes on the surface of the lipid droplets. However, the precise molecular mechanisms for the core-induced formation of lipid droplets remain elusive. Recently, we showed that the expression of the core protein in yeast as a model system could induce lipid droplet formation. In this study, we probed the cellular factors responsible for the formation of core-induced lipid-droplets in yeast cells. We demonstrated that one of the enzymes responsible for triglyceride synthesis, a phospholipid:diacylglycerol acyltransferase (Lro1, is required for the core-induced lipid droplet formation. While core proteins inhibit Lro1 degradation and alter Lro1 localization, the characteristic localization of Lro1 adjacent to the lipid droplets appeared to be responsible for the core-induced lipid droplet formation. RNA virus genomes have evolved using high mutation rates to maintain their ability to replicate. Our observations suggest a functional relationship between the core protein with hepatocytes and yeast cells. The possible interactions between core proteins and the endoplasmic reticulum membrane affect the mobilization of specific proteins.

  19. A study of biodiversity using DSS method and seed storage protein comparison of populations in two species of Achillea L. in the west of Iran

    Directory of Open Access Journals (Sweden)

    Hajar Salehi

    2013-11-01

    Full Text Available Intarspecific and interspecific variations are the main reserves of biodiversity and both are important sources of speciation. On this basis, identifing and recognizing the intra and interspecific variations is important in order to recognition of biodiversity. This research was done to study biodiversity and electrophoresis comparison of seed storage proteins in the populations of the two species of the genus Achillea in Hamadan and Kurdistan provinces using of the method of determination of special station (DSS. For this purpose, 12 and 9 special stations were selected for the species A. tenuifolia and A. biebresteinii using the data published in the related flora. Seed storage proteins were extracted and then studied using electrophoresis techniques (SDS-PAGE. In survey of all special stations, 120 plant species were distinguished as associated species. The results of the floristic data for the both species determined six distinctive groups that indicated the existence of intraspecific diversity in this species. The result of analysis of ecological data and seed storage proteins for the two species was in accordance with the floristic data and showed six distinctive groups. The existence of the bands of no. 4, 5, 8, 12 and 13 in the special stations of A. tenuifolia and the bands of 14, 15 and 16 in the special stations of A. biebresteinii o separated the populations of the species in two quite different and distinctive groups.

  20. Effect of droplet size on the droplet behavior on the heterogeneous surface

    Energy Technology Data Exchange (ETDEWEB)

    Choi, Ho Yeon; Son, Sung Wan; Ha, ManYeong [Pusan National University, Busan (Korea, Republic of); Park, Yong Gap [Pusan National University, Busan (Korea, Republic of)

    2017-06-15

    The characteristics of a three-dimensional hemispherical droplet on a heterogeneous surface were studied using the Lattice Boltzmann method (LBM). The hydrophilic surface has a hydrophobic part at the center. The hemispherical droplets are located at the center of the heterogeneous surface. According to the contact angles of hydrophilic and hydrophobic bottom surfaces, the droplet either separates or reaches a new equilibrium state. The separation time varies according to the change in droplet size, and it affects the status of droplet separation. The droplet separation behavior was investigated by analyzing the velocity vector around the phase boundary line. The shape and separation time of a droplet are determined by the contact angle of each surface. The speed of droplet separation increases as the difference in contact angle increases between the hydrophobic surface and hydrophilic surface. The separation status and the separation time of a droplet are also determined by the change of the droplet size. As the size of the droplet decreases, the effect of surface tension decreases, and the separation time of the droplet also decreases. On the other hand, as the droplet becomes larger, the effect of surface tension increases and the time required for the droplet to separate also increases.

  1. Quantum Nanostructures by Droplet Epitaxy

    Directory of Open Access Journals (Sweden)

    Somsak Panyakeow

    2009-02-01

    Full Text Available Droplet epitaxy is an alternative growth technique for several quantum nanostructures. Indium droplets are distributed randomly on GaAs substrates at low temperatures (120-350'C. Under background pressure of group V elements, Arsenic and Phosphorous, InAs and InP nanostructures are created. Quantum rings with isotropic shape are obtained at low temperature range. When the growth thickness is increased, quantum rings are transformed to quantum dot rings. At high temperature range, anisotropic strain gives rise to quantum rings with square holes and non-uniform ring stripe. Regrowth of quantum dots on these anisotropic quantum rings, Quadra-Quantum Dots (QQDs could be realized. Potential applications of these quantum nanostructures are also discussed.

  2. Shape-Shifting Droplet Networks.

    Science.gov (United States)

    Zhang, T; Wan, Duanduan; Schwarz, J M; Bowick, M J

    2016-03-11

    We consider a three-dimensional network of aqueous droplets joined by single lipid bilayers to form a cohesive, tissuelike material. The droplets in these networks can be programed to have distinct osmolarities so that osmotic gradients generate internal stresses via local fluid flows to cause the network to change shape. We discover, using molecular dynamics simulations, a reversible folding-unfolding process by adding an osmotic interaction with the surrounding environment which necessarily evolves dynamically as the shape of the network changes. This discovery is the next important step towards osmotic robotics in this system. We also explore analytically and numerically how the networks become faceted via buckling and how quasi-one-dimensional networks become three dimensional.

  3. The Effect of pH and High-Pressure Homogenization on Droplet Size

    Directory of Open Access Journals (Sweden)

    Ah Pis Yong

    2017-12-01

    Full Text Available The aims of this study are to revisit the effect of high pressure on homogenization and the influence of pH on the emulsion droplet sizes. The high-pressure homogenization (HPH involves two stages of processing, where the first stage involves in blending the coarse emulsion by a blender, and the second stage requires disruption of the coarse emulsion into smaller droplets by a high-pressure homogenizer. The pressure range in this review is in between 10-500 MPa. The homogenised droplet sizes can be reduced by increasing the homogenization recirculation, and there is a threshold point beyond that by applying pressure only, the size cannot be further reduced. Normally, homogenised emulsions are classified by their degree of kinetic stability. Dispersed phase present in the form of droplets while continuous phase also known as suspended droplets. With a proper homogenization recirculation and pressure, a more kinetically stable emulsion can be produced. The side effects of increasing homogenization pressure are that it can cause overprocessing of the emulsion droplets where the droplet sizes become larger rather than the expected smaller size. This can cause kinetic instability in the emulsion. The droplet size is usually measured by dynamic light scattering or by laser light scattering technique. The type of samples used in this reviews are such as chocolate and vanilla based powders; mean droplet sizes samples; basil oil; tomato; lupin protein; oil; skim milk, soymilk; coconut milk; tomato homogenate; corn; egg-yolk, rapeseed and sunflower; Poly(4-vinylpyridine/silica; and Complex 1 until complex 4 approaches from author case study. A relationship is developed between emulsion size and pH. Results clearly show that lower pH offers smaller droplet of emulsion and the opposite occurs when the pH is increased.

  4. Oxidation of lipid and protein in horse mackerel (Trachurus trachurus) mince and washed minces during processing and storage

    DEFF Research Database (Denmark)

    Eymard, Sylvie; Baron, Caroline; Jacobsen, Charlotte

    2009-01-01

    : M1, M2 and M3, with one, two and three washing steps, respectively. The different products were characterised (i.e. lipid content, protein, water, iron, fatty acid profile and tocopherol content) and analysed for protein and lipid oxidation in order to investigate the impact of the washing steps...... was followed by determination of protein solubility, protein thiol groups and protein carbonyl groups using colorimetric methods as well as western blotting for protein carbonyl groups. Lipid and protein oxidation markers indicated that both lipid and protein oxidation took place during processing...

  5. [Study of the reineta protein modifications (Brama australis), put under freezing and storage to -18 degrees C and -30 degrees C].

    Science.gov (United States)

    Abugoch, Lilian; Quitral, Vilma; Larraín, M Angélica; Vinagre, Julia; Kriukov, Andrei; Chávez, Gloria

    2006-12-01

    The objective of the present work was to study functional and thermal properties of reineta (Brama australis) frozen meat, analysed by water retention capacity (WRC), gel forming capacity (GFC), texture, emulsifying capacity and differential scanning calorimetry (DSC). For this study, reineta fillets were obtained and extracted by the same conditions, and cutted, packaged, frozen and stored at -18 degrees C and -30 degrees C for 7 months. The results obtained, showed that there were no signifficant differences in the responses to thermal treatment for all the specimens. For samples frozen at -18 degrees C and -30 degrees C, the protein contents were 23.5 + 0.0 and 25.4 + 1.0%, respectively. The WRC values were 0.45 + 0.1 and 1.59 +/- 0.0 g water/g protein, respectively. The gel forming capacity was only present in the fresh samples, whereas the frozen stored ones only form protein aggregates. The emulsifying capacity was between 960 and 1400 g oil / g protein, and the storage time increased this value. The miosin denaturation temperature (Td) and denaturation enthalpy (?H), obtained by DSC, fluctuated between 39.2 +/- 0.5 to 44.8 +/- 0.8 degrees C and 1.12 +/- 0.3 to 0.52 +/- 0.2 J/g, respectively. The actina values were between 71.0 +/- 0.6 to 75.3 +/- 0.5 degrees C and between 0.5 +/- 0.1 to 0.7 +/- 0.1 J/g. Cooperativity decreased as the storage time increased. This is showing a certain degree of protein displacement. The values found by thermal analyses showed a direct relationship with the functional properties, both decreasing with storage time.

  6. Can a droplet break up under flow without elongating? Fragmentation of smectic monodisperse droplets

    Science.gov (United States)

    Courbin, L.; Engl, W.; Panizza, P.

    2004-06-01

    We study the fragmentation under shear flow of smectic monodisperse droplets at high volume fraction. Using small angle light scattering and optical microscopy, we reveal the existence of a break-up mechanism for which the droplets burst into daughter droplets of the same size. Surprisingly, this fragmentation process, which is strain controlled and occurs homogeneously in the cell, does not require any transient elongation of the droplets. Systematic experiments as a function of the initial droplet size and the applied shear rate show that the rupture is triggered by an instability of the inner droplet structure.

  7. Impinging Water Droplets on Inclined Glass Surfaces

    Energy Technology Data Exchange (ETDEWEB)

    Armijo, Kenneth Miguel [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Lance, Blake [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Ho, Clifford K. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

    2017-09-01

    Multiphase computational models and tests of falling water droplets on inclined glass surfaces were developed to investigate the physics of impingement and potential of these droplets to self-clean glass surfaces for photovoltaic modules and heliostats. A multiphase volume-of-fluid model was developed in ANSYS Fluent to simulate the impinging droplets. The simulations considered different droplet sizes (1 mm and 3 mm), tilt angles (0°, 10°, and 45°), droplet velocities (1 m/s and 3 m/s), and wetting characteristics (wetting=47° contact angle and non-wetting = 93° contact angle). Results showed that the spread factor (maximum droplet diameter during impact divided by the initial droplet diameter) decreased with increasing inclination angle due to the reduced normal force on the surface. The hydrophilic surface yielded greater spread factors than the hydrophobic surface in all cases. With regard to impact forces, the greater surface tilt angles yielded lower normal forces, but higher shear forces. Experiments showed that the experimentally observed spread factor (maximum droplet diameter during impact divided by the initial droplet diameter) was significantly larger than the simulated spread factor. Observed spread factors were on the order of 5 - 6 for droplet velocities of ~3 m/s, whereas the simulated spread factors were on the order of 2. Droplets were observed to be mobile following impact only for the cases with 45° tilt angle, which matched the simulations. An interesting phenomenon that was observed was that shortly after being released from the nozzle, the water droplet oscillated (like a trampoline) due to the "snapback" caused by the surface tension of the water droplet being released from the nozzle. This oscillation impacted the velocity immediately after the release. Future work should evaluate the impact of parameters such as tilt angle and surface wettability on the impact of particle/soiling uptake and removal to investigate ways that

  8. [Micro-droplet characterization and its application for amino acid detection in droplet microfluidic system].

    Science.gov (United States)

    Yuan, Huiling; Dong, Libing; Tu, Ran; Du, Wenbin; Ji, Shiru; Wang, Qinhong

    2014-01-01

    Recently, the droplet microfluidic system attracts interests due to its high throughput and low cost to detect and screen. The picoliter micro-droplets from droplet microfluidics are uniform with respect to the size and shape, and could be used as monodispensed micro-reactors for encapsulation and detection of single cell or its metabolites. Therefore, it is indispensable to characterize micro-droplet and its application from droplet microfluidic system. We first constructed the custom-designed droplet microfluidic system for generating micro-droplets, and then used the micro-droplets to encapsulate important amino acids such as glutamic acid, phenylalanine, tryptophan or tyrosine to test the droplets' properties, including the stability, diffusivity and bio-compatibility for investigating its application for amino acid detection and sorting. The custom-designed droplet microfluidic system could generate the uniformed micro-droplets with a controllable size between 20 to 50 microm. The micro-droplets could be stable for more than 20 h without cross-contamination or fusion each other. The throughput of detection and sorting of the system is about 600 micro-droplets per minute. This study provides a high-throughput platform for the analysis and screening of amino acid-producing microorganisms.

  9. Proteomic analysis of lettuce seed germination and thermoinhibition by sampling of individual seeds at germination and removal of storage proteins by polyethylene glycol fractionation.

    Science.gov (United States)

    Wang, Wei-Qing; Song, Bin-Yan; Deng, Zhi-Jun; Wang, Yue; Liu, Shu-Jun; Møller, Ian Max; Song, Song-Quan

    2015-04-01

    Germination and thermoinhibition in lettuce (Lactuca sativa 'Jianyexianfeng No. 1') seeds were investigated by a proteomic comparison among dry seeds, germinated seeds at 15°C, at 15°C after imbibition at 25°C for 48 h, or at 25°C in KNO3 (all sampled individually at germination), and ungerminated seeds at 25°C, a thermoinhibitory temperature. Before two-dimensional gel electrophoresis analysis, storage proteins (greater than 50% of total extractable protein) were removed by polyethylene glycol precipitation, which significantly improved the detection of less abundant proteins on two-dimensional gels. A total of 108 protein spots were identified to change more than 2-fold (Pseeds than in ungerminated 25°C seeds. Gene expression of 12 of those proteins correlated well with the protein accumulation. Methionine metabolism, ethylene production, lipid mobilization, cell elongation, and detoxification of aldehydes were revealed to be potentially related to lettuce seed germination and thermoinhibition. Accumulation of three proteins and expression of five genes participating in the mevalonate (MVA) pathway of isoprenoid biosynthesis correlated positively with seed germinability. Inhibition of this pathway by lovastatin delayed seed germination and increased the sensitivity of germination to abscisic acid. MVA pathway-derived products, cytokinins, partially reversed the lovastatin inhibition of germination and released seed thermoinhibition at 25°C. We conclude that the MVA pathway for isoprenoid biosynthesis is involved in lettuce seed germination and thermoinhibition. © 2015 American Society of Plant Biologists. All Rights Reserved.

  10. Systems, methods, and products for graphically illustrating and controlling a droplet actuator

    Science.gov (United States)

    Paik, Philip Y. (Inventor); Pollack, Michael G. (Inventor); Sturmer, Ryan A. (Inventor); Smith, Gregory F. (Inventor); Brafford, Keith R. (Inventor); Pamula, Vamsee K. (Inventor)

    2010-01-01

    Systems for controlling a droplet microactuator are provided. According to one embodiment, a system is provided and includes a controller, a droplet microactuator electronically coupled to the controller, and a display device displaying a user interface electronically coupled to the controller, wherein the system is programmed and configured to permit a user to effect a droplet manipulation by interacting with the user interface. According to another embodiment, a system is provided and includes a processor, a display device electronically coupled to the processor, and software loaded and/or stored in a storage device electronically coupled to the controller, a memory device electronically coupled to the controller, and/or the controller and programmed to display an interactive map of a droplet microactuator. According to yet another embodiment, a system is provided and includes a controller, a droplet microactuator electronically coupled to the controller, a display device displaying a user interface electronically coupled to the controller, and software for executing a protocol loaded and/or stored in a storage device electronically coupled to the controller, a memory device electronically coupled to the controller, and/or the controller.

  11. Experimental test of liquid droplet radiator performance

    Science.gov (United States)

    Mattick, A. T.; Simon, M. A.

    The liquid droplet radiator (LDR) is a heat rejection system for space power systems wherein an array of heated liquid droplets radiates energy directly to space. The use of submillimeter droplets provides large radiating area-to-mass ratio, resulting in radiator systems which are several times lighter than conventional solid surface radiators. An experiment is described in which the power radiated by an array of 2300 streams of silicone oil droplets is measured to test a previously developed theory of the LDR radiation process. This system would be capable of rejecting several kW of heat in space. Furthermore, it would be suitable as a modular unit of an LDR designed for 100-kW power levels. The experiment provided confirmation of the theoretical dependence of droplet array emissivity on optical depth. It also demonstrated the ability to create an array of more than 1000 droplet streams having a divergence less than 1 degree.

  12. A parameterization of cloud droplet nucleation

    International Nuclear Information System (INIS)

    Ghan, S.J.; Chuang, C.; Penner, J.E.

    1993-01-01

    Droplet nucleation is a fundamental cloud process. The number of aerosols activated to form cloud droplets influences not only the number of aerosols scavenged by clouds but also the size of the cloud droplets. Cloud droplet size influences the cloud albedo and the conversion of cloud water to precipitation. Global aerosol models are presently being developed with the intention of coupling with global atmospheric circulation models to evaluate the influence of aerosols and aerosol-cloud interactions on climate. If these and other coupled models are to address issues of aerosol-cloud interactions, the droplet nucleation process must be adequately represented. Here we introduce a droplet nucleation parametrization that offers certain advantages over the popular Twomey (1959) parameterization

  13. Water Entry by a Train of Droplets

    Science.gov (United States)

    Ohl, Claus-Dieter; Huang, Xin; Chan, Chon U.; Frommhold, Philipp Erhard; Lippert, Alexander

    2014-11-01

    The impact of single droplets on a deep pool is a well-studied phenomenon which reveals reach fluid mechanics. Lesser studied is the impact of a train of droplet and the accompanied formation of largely elongated cavities, in particular for well controlled droplets. The droplets with diameters of 20-40 μm and velocities of approx. 20 m/s are generated with a piezo-actuated nozzle at rates of 200-300 kHz. Individual droplets are selected by electric charging and deflection and the impact is visualized with stroboscopic photography and high-speed videos. We study in particular the formation and shape of the cavity as by varying the number of droplets from one to 64. The cavities reach centimetres in length with lateral diameters of the order of 100 of micrometres.

  14. Molecular cloning and characterization of Polygalacturonase-Inhibiting Protein and Cinnamoyl-Coa Reductase genes and their association with fruit storage conditions in blueberry (Vaccinium corymbosum)

    KAUST Repository

    Khraiwesh, Basel

    2013-05-13

    Blueberry is a widely grown and easily perishable fruit crop. An efficient post-harvest handling is critical, and for that purpose gene technology methods have been part of ongoing programmes to improve crops with high food values such as blueberry. Here we report the isolation, cloning, characterization and differential expression levels of two cDNAs encoding Polygalacturonase-Inhibitor Protein (PGIP) and Cinnamoyl-Coa Reductase (CCR) from blueberry fruits in relation to various storage conditions. The open reading frame of PGIP and CCR encodes a polypeptide of 329 and 347 amino acids, respectively. To assess changes in the expression of blueberry PGIP and CCR after harvest, a storage trial was initiated. The northern blots hybridization showed a clear differential expression level of PGIP and CCR between freshly harvested and stored fruits as well as between fruits stored under various storage conditions. Although the prospects of exploiting such a strategy for crop improvement are limited, the results provide further insight into the control of the quality over the storage period at the molecular level.

  15. Molecular cloning and characterization of Polygalacturonase-Inhibiting Protein and Cinnamoyl-Coa Reductase genes and their association with fruit storage conditions in blueberry (Vaccinium corymbosum)

    KAUST Repository

    Khraiwesh, Basel; Harb, Jamil; Qudeimat, Enas

    2013-01-01

    Blueberry is a widely grown and easily perishable fruit crop. An efficient post-harvest handling is critical, and for that purpose gene technology methods have been part of ongoing programmes to improve crops with high food values such as blueberry. Here we report the isolation, cloning, characterization and differential expression levels of two cDNAs encoding Polygalacturonase-Inhibitor Protein (PGIP) and Cinnamoyl-Coa Reductase (CCR) from blueberry fruits in relation to various storage conditions. The open reading frame of PGIP and CCR encodes a polypeptide of 329 and 347 amino acids, respectively. To assess changes in the expression of blueberry PGIP and CCR after harvest, a storage trial was initiated. The northern blots hybridization showed a clear differential expression level of PGIP and CCR between freshly harvested and stored fruits as well as between fruits stored under various storage conditions. Although the prospects of exploiting such a strategy for crop improvement are limited, the results provide further insight into the control of the quality over the storage period at the molecular level.

  16. Heteroaggregation of lipid droplets coated with sodium caseinate and lactoferrin.

    Science.gov (United States)

    de Figueiredo Furtado, Guilherme; Michelon, Mariano; de Oliveira, Davi Rocha Bernardes; da Cunha, Rosiane Lopes

    2016-11-01

    Formation and characterization of droplet heteroaggregates were investigated by mixing two emulsions previously stabilized by proteins oppositely charged. Emulsions were composed of 5vol.% of sunflower oil and 95vol.% of sodium caseinate or lactoferrin aqueous dispersions. They were produced using ultrasound with fixed power (300W) and sonication time (6min). Different volume ratios (0-100%) of sodium caseinate-stabilized emulsion (droplet diameter around 1.75μm) to lactoferrin-stabilized emulsion (droplet diameter around 1.55μm) were mixed under conditions that both proteins showed opposite charges (pH7). Influence of ionic strength (0-400mM NaCl) on the heteroaggregates stability was also evaluated. Creaming stability, zeta potential, microstructure, mean particle diameter and rheological properties of the heteroaggregates were measured. These properties depended on the volume ratio (0-100%) of sodium caseinate to lactoferrin-stabilized emulsion (C:L) and the ionic strength. In the absence of salt, different zeta potential values were obtained, rheological properties (viscosity and elastic moduli) were improved and the largest heteroaggregates were formed at higher content of lactoferrin-stabilized emulsion (60-80%). The system containing 40 and 60vol.% of sodium caseinate and lactoferrin stabilized emulsion, respectively, presented good stability against phase separation besides showing enhanced rheological and size properties due to extensive droplets aggregation. Phase separation was observed only in the absence of sodium caseinate, demonstrating the higher susceptibility of lactoferrin to NaCl. The heteroaggregates produced may be useful functional agents for texture modification and controlled release since different rheological properties and sizes can be achieved depending on protein concentrations. Copyright © 2016 Elsevier Ltd. All rights reserved.

  17. Identification and characterization of finger millet OPAQUE2 transcription factor gene under different nitrogen inputs for understanding their role during accumulation of prolamin seed storage protein.

    Science.gov (United States)

    Gaur, Vikram Singh; Kumar, Lallan; Gupta, Supriya; Jaiswal, J P; Pandey, Dinesh; Kumar, Anil

    2018-03-01

    In this study, we report the isolation and characterization of the mRNA encoding OPAQUE2 (O2) like TF of finger millet (FM) ( Eleusine coracana) ( EcO2 ). Full-length EcO2 mRNA was isolated using conserved primers designed by aligning O2 mRNAs of different cereals followed by 3' and 5' RACE (Rapid Amplification of cDNA Ends). The assembled full-length EcO2 mRNA was found to contain an ORF of 1248-nt coding the 416 amino acids O2 protein. Domain analysis revealed the presence of the BLZ and bZIP-C domains which is a characteristic feature of O2 proteins. Phylogenetic analysis of EcO2 protein with other bZIP proteins identified using finger millet transcriptome data and O2 proteins of other cereals showed that EcO2 shared high sequence similarity with barley BLZ1 protein. Transcripts of EcO2 were detected in root, stem, leaves, and seed development stages. Furthermore, to investigate nitrogen responsiveness and the role of EcO2 in regulating seed storage protein gene expression, the expression profiles of EcO2 along with an α-prolamin gene were studied during the seed development stages of two FM genotypes (GE-3885 and GE-1437) differing in grain protein content (13.8 and 6.2%, respectively) grown under increasing nitrogen inputs. Compared to GE-1437, the EcO2 was relatively highly expressed during the S2 stage of seed development which further increased as nitrogen input was increased. The Ecα - prolamin gene was strongly induced in the high protein genotype (GE-3885) at all nitrogen inputs. These results indicate the presence of nitrogen responsiveness regulatory elements which might play an important role in accumulating protein in FM genotypes through modulating EcO2 expression by sensing plant nitrogen status.

  18. Preventing droplet deformation during dielectrophoretic centering of a compound emulsion droplet

    Science.gov (United States)

    Randall, Greg; Blue, Brent

    2012-11-01

    Compound droplets, or droplets-within-droplets, are traditionally key components in applications ranging from drug delivery to the food industry. Presently, millimeter-sized compound droplets are precursors for shell targets in inertial fusion energy work. However, a key constraint in target fabrication is a uniform shell wall thickness, which in turn requires a centered core droplet in the compound droplet precursor. Previously, Bei et al. (2009, 2010) have shown that compound droplets could be centered in a static fluid using an electric field of 0.7 kV/cm at 20 MHz. Randall et al. (2012) developed a process to center the core of a moving compound droplet, though the ~kV/cm field induced small (fluid mechanics and interfacial rheology perspective and we discuss the effective interfacial charge from an emulsifier and its impact on centering. Work funded by General Atomics Internal R&D.

  19. Evaporation of nanofluid droplet on heated surface

    Directory of Open Access Journals (Sweden)

    Yeung Chan Kim

    2015-04-01

    Full Text Available In this study, an experiment on the evaporation of nanofluid sessile droplet on a heated surface was conducted. A nanofluid of 0.5% volumetric concentration mixed with 80-nm-sized CuO powder and pure water were used for experiment. Droplet was applied to the heated surface, and images of the evaporation process were obtained. The recorded images were analyzed to find the volume, diameter, and contact angle of the droplet. In addition, the evaporative heat transfer coefficient was calculated from experimental result. The results of this study are summarized as follows: the base diameter of the droplet was maintained stably during the evaporation. The measured temperature of the droplet was increased rapidly for a very short time, then maintained constantly. The nanofluid droplet was evaporated faster than the pure water droplet under the experimental conditions of the same initial volume and temperature, and the average evaporative heat transfer coefficient of the nanofluid droplet was higher than that of pure water. We can consider the effects of the initial contact angle and thermal conductivity of nanofluid as the reason for this experimental result. However, the effect of surface roughness on the evaporative heat transfer of nanofluid droplet appeared unclear.

  20. Droplet size in a rectangular Venturi scrubber

    OpenAIRE

    Costa, M. A. M.; Henrique, P. R.; Gonçalves, J. A. S.; Coury, J.R.

    2004-01-01

    The Venturi scrubber is a device which uses liquid in the form of droplets to efficiently remove fine particulate matter from gaseous streams. Droplet size is of fundamental importance for the scrubber performance. In the present experimental study, a laser diffraction technique was used in order to measure droplet size in situ in a Venturi scrubber with a rectangular cross section. Droplet size distribution was measured as a function of gas velocity (58.3 to 74.9 m/s), liquid-to-gas ratio (0...

  1. Janus droplet as a catalytic micromotor

    Science.gov (United States)

    Shklyaev, Sergey

    2015-06-01

    Self-propulsion of a Janus droplet in a solution of surfactant, which reacts on a half of a drop surface, is studied theoretically. The droplet acts as a catalytic motor creating a concentration gradient, which generates its surface-tension-driven motion; the self-propulsion speed is rather high, 60 μ \\text{m/s} and more. This catalytic motor has several advantages over other micromotors: simple manufacturing, easily attained neutral buoyancy. In contrast to a single-fluid droplet, which demonstrates a self-propulsion as a result of symmetry breaking instability, for the Janus one no stability threshold exists; hence, the droplet radius can be scaled down to micrometers.

  2. Calculation and measurement of fog droplet size

    International Nuclear Information System (INIS)

    Laali, A.R.; Courant, J.J.; Kleitz, A.

    1991-01-01

    This paper outlines the elements involved in calculation and measurement of fog droplet size in steam turbines. The condensation calculations are performed for a 600 MW LP fossil fired, and for a 900 MW LP nuclear turbine. A simplified method based on classical condensation theory is used for these calculations. The fog droplet size measurement are carried out downstream of the last moving blades of these turbines in order to validate the program. The comparison between the results could lead to a better understanding of the condensation process in steam turbines. Some large droplet (re-entrained droplet) measurements are also taken using a microvideo probe

  3. Colliding droplets: a short film presentation

    International Nuclear Information System (INIS)

    Hendricks, C.D.

    1981-01-01

    A series of experiments were performed in which liquid droplets were caused to collide. Impact velocities to several meters per second and droplet diameters up to 600 micrometers were used. The impact parameters in the collisions vary from zero to greater than the sum of the droplet radii. Photographs of the collisions were taken with a high speed framing camera in order to study the impacts and subsequent behavior of the droplets. The experiments will be discussed and a short movie film presentation of some of the impacts will be shown

  4. Shock wave-induced evaporation of water droplets in a gas-droplet mixture 646

    NARCIS (Netherlands)

    Goossens, H.W.J.; Cleijne, J.W.; Smolders, H.J.; Dongen, van M.E.H.

    1988-01-01

    A model is presented for the droplet evaporation process induced by a shock wave propagating in a fog. The model is based on the existence of a quasi-steady wet bulb state of the droplets during evaporation. It is shown that for moderate shock strength, Ma = <2,=" and=" droplet=" radii=" in=" the="

  5. ORFer--retrieval of protein sequences and open reading frames from GenBank and storage into relational databases or text files.

    Science.gov (United States)

    Büssow, Konrad; Hoffmann, Steve; Sievert, Volker

    2002-12-19

    Functional genomics involves the parallel experimentation with large sets of proteins. This requires management of large sets of open reading frames as a prerequisite of the cloning and recombinant expression of these proteins. A Java program was developed for retrieval of protein and nucleic acid sequences and annotations from NCBI GenBank, using the XML sequence format. Annotations retrieved by ORFer include sequence name, organism and also the completeness of the sequence. The program has a graphical user interface, although it can be used in a non-interactive mode. For protein sequences, the program also extracts the open reading frame sequence, if available, and checks its correct translation. ORFer accepts user input in the form of single or lists of GenBank GI identifiers or accession numbers. It can be used to extract complete sets of open reading frames and protein sequences from any kind of GenBank sequence entry, including complete genomes or chromosomes. Sequences are either stored with their features in a relational database or can be exported as text files in Fasta or tabulator delimited format. The ORFer program is freely available at http://www.proteinstrukturfabrik.de/orfer. The ORFer program allows for fast retrieval of DNA sequences, protein sequences and their open reading frames and sequence annotations from GenBank. Furthermore, storage of sequences and features in a relational database is supported. Such a database can supplement a laboratory information system (LIMS) with appropriate sequence information.

  6. Wild blueberry polyphenol-protein food ingredients produced by three drying methods: Comparative physico-chemical properties, phytochemical content, and stability during storage.

    Science.gov (United States)

    Correia, Roberta; Grace, Mary H; Esposito, Debora; Lila, Mary Ann

    2017-11-15

    Particulate colloidal aggregate food ingredients were prepared by complexing wheat flour, chickpea flour, coconut flour and soy protein isolate with aqueous wild blueberry pomace extracts, then spray drying, freeze drying, or vacuum oven drying to prepare dry, flour-like matrices. Physico-chemical attributes, phytochemical content and stability during storage were compared. Eighteen anthocyanins peaks were identified for samples. Spray dried matrices produced with soy protein isolate had the highest concentration of polyphenols (156.2mg GAE/g) and anthocyanins (13.4mg/g) and the most potent DPPH scavenging activity (714.1μmolesTE/g). Spray dried blueberry polyphenols complexed with protein were protected from degradation during 16weeks at 4°C and 20°C. Soy protein isolate more efficiently captured and stabilized wild blueberry pomace phytochemicals than other protein sources. Overall, spray drying the blueberry extracts complexed with protein proved to be an environment-friendly strategy to produce stable functional ingredients with multiple applications for the food industry. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. Energy Storage.

    Science.gov (United States)

    Eaton, William W.

    Described are technological considerations affecting storage of energy, particularly electrical energy. The background and present status of energy storage by batteries, water storage, compressed air storage, flywheels, magnetic storage, hydrogen storage, and thermal storage are discussed followed by a review of development trends. Included are…

  8. Seipin is required for converting nascent to mature lipid droplets

    DEFF Research Database (Denmark)

    Wang, Huajin; Becuwe, Michel; Housden, Benjamin E

    2016-01-01

    How proteins control the biogenesis of cellular lipid droplets (LDs) is poorly understood. Using Drosophila and human cells, we show here that seipin, an ER protein implicated in LD biology, mediates a discrete step in LD formation-the conversion of small, nascent LDs to larger, mature LDs. Seipin...... leading to the giant LDs characteristic of seipin deficiency. Our studies identify a discrete step of LD formation, namely the conversion of nascent LDs to mature LDs, and define a molecular role for seipin in this process, most likely by acting at ER-LD contact sites to enable lipid transfer to nascent...

  9. Effect of storage temperature on survival and recovery of thermal and extrusion injured Escherichia coli K-12 in whey protein concentrate and corn meal.

    Science.gov (United States)

    Ukuku, Dike O; Mukhopadhyay, Sudarsan; Onwulata, Charles

    2013-01-01

    Previously, we reported inactivation of Escherichia coli populations in corn product (CP) and whey protein product (WPP) extruded at different temperatures. However, information on the effect of storage temperatures on injured bacterial populations was not addressed. In this study, the effect of storage temperatures on the survival and recovery of thermal death time (TDT) disks and extrusion injured E. coli populations in CP and WPP was investigated. CP and WPP inoculated with E. coli bacteria at 7.8 log(10) CFU/g were conveyed separately into the extruder with a series 6300 digital type T-35 twin screw volumetric feeder set at a speed of 600 rpm and extruded at 35°C, 55°C, 75°C, and 95°C, or thermally treated with TDT disks submerged into water bath set at 35°C, 55°C, 75°C, and 95°C for 120 s. Populations of surviving bacteria including injured cells in all treated samples were determined immediately and every day for 5 days, and up to 10 days for untreated samples during storage at 5°C, 10°C, and 23°C. TDT disks treatment at 35°C and 55°C did not cause significant changes in the population of the surviving bacteria including injured populations. Extrusion treatment at 35°C and 55°C led to significant (pagar plates. The results of this study showed that further inactivation of the injured populations occurred during storage at 5°C for 5 days suggesting the need for immediate storage of 75°C extruded CP and WPP at 5°C for at least 24 h to enhance their microbial safety.

  10. Shear-driven redistribution of surfactant affects enzyme activity in well-mixed femtoliter droplets.

    Science.gov (United States)

    Liu, Yu; Jung, Seung-Yong; Collier, C Patrick

    2009-06-15

    We developed a microfluidic platform for splitting well-mixed, femtoliter-volume droplets from larger water-in-oil plugs, where the sizes of the daughter droplets were not limited by channel width. These droplets were separated from mother plugs at a microfabricated T-junction, which enabled the study of how increased confinement affected enzyme kinetics in droplets 4-10 microm in diameter. Initial rates for enzyme catalysis in the mother plugs and the largest daughter drops were close to the average bulk rate, while the rates in smaller droplets decreased linearly with increasing surface to volume ratio. Rates in the smallest droplets decreased by a factor of 4 compared to the bulk rate. Traditional methods for detecting nonspecific adsorption at the water-oil interface were unable to detect evidence of enzyme adsorption, including pendant drop tensiometry, laser scanning confocal microscopy of drops containing labeled proteins in microemulsions, and epifluorescence microscopy of plugs and drops generated on-chip. We propose the slowing of enzyme reaction kinetics in the smaller droplets was the result of increased adsorption and inactivation of enzymes at the water-oil interface arising from transient interfacial shear stresses imparted on the daughter droplets as they split from the mother plugs and passed through the constricted opening of the T-junction. Such stresses are known to modulate the interfacial area and density of surfactant molecules that can passivate the interface. Bright field images of the splitting processes at the junction indicate that these stresses scaled with increasing surface to volume ratios of the droplets but were relatively insensitive to the average flow rate of plugs upstream of the junction.

  11. Effects of heat stress and probiotic supplementation on protein functionality and oxidative stability of ground chicken leg meat during display storage.

    Science.gov (United States)

    Kim, Hyun-Wook; Kim, Ji-Han; Yan, Feifei; Cheng, Heng-Wei; Brad Kim, Yuan H

    2017-12-01

    The present study aimed to evaluate the effects of heat stress and probiotic supplementation on protein functionality and oxidative stability of ground chicken leg during display storage. Two hundred and forty, 1-day-old male chicks (5 birds per pen) were subjected to four treatments in a 2 (thermoneutral condition at 21 °C and cyclic heat stress at 32-21-32 °C for 10 h day -1 ) × 2 (regular diet with 0 or 0.25 g kg -1 Bacillus subtilis) factorial design. Chickens were harvested at day 46, and pairs of whole legs were collected at 1 day postmortem. The chicken legs were deboned, ground, tray-packaged with oxygen-permeable film, and displayed for 3 days. Heat stress and probiotic supplementation had no impact on pH, water-holding capacity, color, protein functionality, lipid lipolysis and lipid/protein oxidation stability (P > 0.05). Display storage increased the pH and lipid oxidation of ground chicken legs (P chicken leg meat. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  12. Droplet Vaporization In A Levitating Acoustic Field

    Science.gov (United States)

    Ruff, G. A.; Liu, S.; Ciobanescu, I.

    2003-01-01

    Combustion experiments using arrays of droplets seek to provide a link between single droplet combustion phenomena and the behavior of complex spray combustion systems. Both single droplet and droplet array studies have been conducted in microgravity to better isolate the droplet interaction phenomena and eliminate or reduce the effects of buoyancy-induced convection. In most experiments involving droplet arrays, the droplets are supported on fibers to keep them stationary and close together before the combustion event. The presence of the fiber, however, disturbs the combustion process by introducing a source of heat transfer and asymmetry into the configuration. As the number of drops in a droplet array increases, supporting the drops on fibers becomes less practical because of the cumulative effect of the fibers on the combustion process. To eliminate the effect of the fiber, several researchers have conducted microgravity experiments using unsupported droplets. Jackson and Avedisian investigated single, unsupported drops while Nomura et al. studied droplet clouds formed by a condensation technique. The overall objective of this research is to extend the study of unsupported drops by investigating the combustion of well-characterized drop clusters in a microgravity environment. Direct experimental observations and measurements of the combustion of droplet clusters would provide unique experimental data for the verification and improvement of spray combustion models. In this work, the formation of drop clusters is precisely controlled using an acoustic levitation system so that dilute, as well as dense clusters can be created and stabilized before combustion in microgravity is begun. While the low-gravity test facility is being completed, tests have been conducted in 1-g to characterize the effect of the acoustic field on the vaporization of single and multiple droplets. This is important because in the combustion experiment, the droplets will be formed and

  13. Regulation of the endoplasmic reticulum calcium storage during the unfolded protein response--significance in tissue ischemia?

    DEFF Research Database (Denmark)

    Treiman, Marek

    2002-01-01

    for the protein folding pathway require Ca(2+) binding for their activity. A number of factors, including Ca(2+) depletion, may interfere with the folding pathway within the ER, with a potential for cell injury through an accumulation of malfolded protein aggregates. The Unfolded Protein Response involves...... a transcriptional upregulation of a number of the ER-resident folding helper proteins and becomes triggered when the folding pathway is blocked. To be effective, these upregulated proteins require a sufficient supply of Ca(2+) cofactor within the ER lumen. In tissue ischemia, where the availablity of this cofactor...

  14. Iron storage proteins are essential for the survival and pathogenesis of Mycobacterium tuberculosis in THP-1 macrophages and the guinea pig model of infection.

    Science.gov (United States)

    Reddy, P Vineel; Puri, Rupangi Verma; Khera, Aparna; Tyagi, Anil K

    2012-02-01

    Iron is one of the crucial elements required for the growth of Mycobacterium tuberculosis. However, excess free iron becomes toxic for the cells because it catalyzes the production of reactive oxygen radicals, leading to oxidative damage. Hence, it is essential for the pathogen to have the ability to store intracellular iron in an iron-rich environment and utilize it under iron depletion. M. tuberculosis has two iron storage proteins, namely BfrA (Rv1876; a bacterioferritin) and BfrB (Rv3841; a ferritin-like protein). However, the demonstration of biological significance requires the disruption of relevant genes and the evaluation of the resulting mutant for its ability to survive in the host and cause disease. In this study, we have disrupted bfrA and bfrB of M. tuberculosis and demonstrated that these genes are crucial for the storage and supply of iron for the growth of bacteria and to withstand oxidative stress in vitro. In addition, the bfrA bfrB double mutant (H37Rv ΔbfrA ΔbfrB) exhibited a marked reduction in its ability to survive inside human macrophages. Guinea pigs infected with H37Rv ΔbfrA ΔbfrB exhibited a marked diminution in the dissemination of the bacilli to spleen compared to that of the parental strain. Moreover, guinea pigs infected with H37Rv ΔbfrA ΔbfrB exhibited significantly reduced pathological damage in spleen and lungs compared to that of animals infected with the parental strain. Our study clearly demonstrates the importance of these iron storage proteins in the survival and pathogenesis of M. tuberculosis in the host and establishes them as attractive targets for the development of new inhibitors against mycobacterial infections.

  15. Slip of Spreading Viscoplastic Droplets.

    Science.gov (United States)

    Jalaal, Maziyar; Balmforth, Neil J; Stoeber, Boris

    2015-11-10

    The spreading of axisymmetric viscoplastic droplets extruded slowly on glass surfaces is studied experimentally using shadowgraphy and swept-field confocal microscopy. The microscopy furnishes vertical profiles of the radial velocity using particle image velocimetry (PIV) with neutrally buoyant tracers seeded in the fluid. Experiments were conducted for two complex fluids: aqueous solutions of Carbopol and xanthan gum. On untreated glass surfaces, PIV demonstrates that both fluids experience a significant amount of effective slip. The experiments were repeated on glass that had been treated to feature positive surface charges, thereby promoting adhesion between the negatively charged polymeric constituents of the fluids and the glass surface. The Carbopol and xanthan gum droplets spread more slowly on the treated surface and to a smaller radial distance. PIV demonstrated that this reduced spreading was associated with a substantial reduction in slip. For Carbopol, the effective slip could be eliminated entirely to within the precision of the PIV measurements; the reduction in slip was less effective for xanthan gum, with a weak slip velocity remaining noticeable.

  16. Experimental test of liquid droplet radiator performance

    International Nuclear Information System (INIS)

    Mattick, A.T.; Simon, M.A.

    1986-01-01

    This liquid droplet radiator (LDR) is evolving rapidly as a lightweight system for heat rejection in space power systems. By using recirculating free streams of submillimeter droplets to radiate waste energy directly to space, the LDR can potentially be an order of magnitude lighter than conventional radiator systems which radiate from solid surfaces. The LDR is also less vulnerable to micrometeoroid damage than are conventional radiators, and it has a low transport volume. Three major development issues of this new heat rejection system are the ability to direct the droplet streams with sufficient precision to avoid fluid loss, radiative performance of the array of droplet streams which comprise the radiating elements of the LDR, and the efficacy of the droplet stream collector, again with respect to fluid loss. This paper reports experimental results bearing on the first two issues - droplet aiming in a multikilowatt-sized system, and radiated power from a large droplet array. Parallel efforts on droplet collection and LDR system design are being pursued by several research groups

  17. Multicomponent droplet vaporization in a convecting environment

    International Nuclear Information System (INIS)

    Megaridis, C.M.; Sirignano, W.A.

    1990-01-01

    In this paper a parametric study of the fundamental exchange processes for energy, mass and momentum between the liquid and gas phases of multicomponent liquid vaporizing droplets is presented. The model, which examines an isolated, vaporizing, multicomponent droplet in an axisymmetric, convecting environment, considers the different volatilities of the liquid components, the alteration of the liquid-phase properties due to the spatial/temporal variations of the species concentrations and also the effects of multicomponent diffusion. In addition, the model accounts for variable thermophysical properties, surface blowing and droplet surface regression due to vaporization, transient droplet heating with internal liquid circulation, and finally droplet deceleration with respect to the free flow due to drag. The numerical calculation employs finite-difference techniques and an iterative solution procedure that provides time-varying spatially-resolved data for both phases. The effects of initial droplet composition, ambient temperature, initial Reynolds number (based on droplet diameter), and volatility differential between the two liquid components are investigated for a liquid droplet consisting of two components with very different volatilities. It is found that mixtures with higher concentration of the less volatile substance actually vaporize faster on account of intrinsically higher liquid heating rates

  18. Proteomic Analysis of Lettuce Seed Germination and Thermoinhibition by Sampling of Individual Seeds at Germination and Removal of Storage Proteins by Polyethylene Glycol Fractionation1

    Science.gov (United States)

    Song, Bin-Yan; Deng, Zhi-Jun; Wang, Yue; Liu, Shu-Jun; Møller, Ian Max; Song, Song-Quan

    2015-01-01

    Germination and thermoinhibition in lettuce (Lactuca sativa ‘Jianyexianfeng No. 1’) seeds were investigated by a proteomic comparison among dry seeds, germinated seeds at 15°C, at 15°C after imbibition at 25°C for 48 h, or at 25°C in KNO3 (all sampled individually at germination), and ungerminated seeds at 25°C, a thermoinhibitory temperature. Before two-dimensional gel electrophoresis analysis, storage proteins (greater than 50% of total extractable protein) were removed by polyethylene glycol precipitation, which significantly improved the detection of less abundant proteins on two-dimensional gels. A total of 108 protein spots were identified to change more than 2-fold (P lettuce seed germination and thermoinhibition. Accumulation of three proteins and expression of five genes participating in the mevalonate (MVA) pathway of isoprenoid biosynthesis correlated positively with seed germinability. Inhibition of this pathway by lovastatin delayed seed germination and increased the sensitivity of germination to abscisic acid. MVA pathway-derived products, cytokinins, partially reversed the lovastatin inhibition of germination and released seed thermoinhibition at 25°C. We conclude that the MVA pathway for isoprenoid biosynthesis is involved in lettuce seed germination and thermoinhibition. PMID:25736209

  19. Fast electric control of the droplet size in a microfluidic T-junction droplet generator

    Science.gov (United States)

    Shojaeian, Mostafa; Hardt, Steffen

    2018-05-01

    The effect of DC electric fields on the generation of droplets of water and xanthan gum solutions in sunflower oil at a microfluidic T-junction is experimentally studied. The electric field leads to a significant reduction of the droplet diameter, by about a factor of 2 in the case of water droplets. The droplet size can be tuned by varying the electric field strength, an effect that can be employed to produce a stream of droplets with a tailor-made size sequence. Compared to the case of purely hydrodynamic droplet production without electric fields, the electric control has about the same effect on the droplet size if the electric stress at the liquid/liquid interface is the same as the hydrodynamic stress.

  20. Droplet behavior analysis in consideration of droplet entrainment from liquid film in annular dispersed flow

    International Nuclear Information System (INIS)

    Matsuura, Keizo; Otake, Hiroshi; Kataoka, Isao; Serizawa, Akimi

    2000-01-01

    A method of droplet behavior simulation in an annular dispersed flow has been developed. In this method, both droplet deposition and entrainment from liquid film are considered. The Lagrangian method and stochastic model are used to analyze droplet diffusion and deposition behavior in a turbulent flow, and droplet entrainment from liquid film is calculated by an entrainment correlation. For the verification of this method, Gill's experiment is analyzed, in which the transition from annular flow with no entrainment to equilibrium annular dispersed flow was observed. Analysis results can also show the similar transition tendency. The experimental results of radial distribution of droplet mass flux are compared with analysis results. The agreement is good for low liquid flow rate, but entrainment rate must be adjusted for high liquid flow rate, in which gas turbulence is thought to be modified by high droplet density. In future work the effect of high droplet density on turbulence should be considered. (author)

  1. Preparation and nucleation of spherical metallic droplet

    Directory of Open Access Journals (Sweden)

    Bing-ge Zhao

    2015-03-01

    Full Text Available The preparation and solidification of metallic droplets attract more and more attention for their significance in both engineering and scientific fields. In this paper, the preparation and characterization of Sn-based alloy droplets using different methods such as atomization and consumable electrode direct current arc (CDCA technique are reviewed. The morphology and structure of these droplets were determined by optical microscopy, X-ray diffraction (XRD and scanning electron microscopy (SEM. The solidification behavior of single droplet was systematically studied by means of scanning calorimetry (DSC, and the nucleation kinetics was also calculated. In particular, the development of fast scanning calorimetry (FSC made it possible to investigate the evolution of undercooling under ultrafast but controllable heating and cooling conditions. The combination of CDCA technique and FSC measurements opens up a new door for quantitative studies on droplet solidification, which is accessible to demonstrate some theories by experiments.

  2. New models for droplet heating and evaporation

    KAUST Repository

    Sazhin, Sergei S.

    2013-02-01

    A brief summary of new models for droplet heating and evaporation, developed mainly at the Sir Harry Ricardo Laboratory of the University of Brighton during 2011-2012, is presented. These are hydrodynamic models for mono-component droplet heating and evaporation, taking into account the effects of the moving boundary due to evaporation, hydrodynamic models of multi-component droplet heating and evaporation, taking and not taking into account the effects of the moving boundary, new kinetic models of mono-component droplet heating and evaporation, and a model for mono-component droplet evaporation, based on molecular dynamics simulation. The results, predicted by the new models are compared with experimental data and the prehctions of the previously developed models where possible. © 2013 Asian Network for Scientific Information.

  3. Settling of fixed erythrocyte suspension droplets

    Science.gov (United States)

    Omenyi, S. N.; Snyder, R. S.

    1983-01-01

    It is pointed out that when particles behave collectively rather than individually, the fractionation of micron-size particles on the basis of size, density, and surface characteristics by centrifugation and electrophoresis is hindered. The formation and sedimentation of droplets containing particles represent an extreme example of collective behavior and pose a major problem for these separation methods when large quantities of particles need to be fractionated. Experiments are described that measure droplet sizes and settling rates for a variety of particles and droplets. Expressions relating the particle concentration in a drop to measurable quantities of the fluids and particles are developed. The number of particles in each droplet is then estimated, together with the effective droplet density. Red blood cells from different animals fixed in glutaraldehyde provide model particle groups.

  4. Droplet size in a rectangular Venturi scrubber

    Directory of Open Access Journals (Sweden)

    M. A. M. Costa

    2004-06-01

    Full Text Available The Venturi scrubber is a device which uses liquid in the form of droplets to efficiently remove fine particulate matter from gaseous streams. Droplet size is of fundamental importance for the scrubber performance. In the present experimental study, a laser diffraction technique was used in order to measure droplet size in situ in a Venturi scrubber with a rectangular cross section. Droplet size distribution was measured as a function of gas velocity (58.3 to 74.9 m/s, liquid-to-gas ratio (0.07 to 0.27 l/m³, and distance from liquid injection point (64 to 173 mm. It was found that all these variables significantly affect droplet size. The results were compared with the predictions from correlations found in the literature.

  5. Effects of Tannic Acid on Lipid and Protein Oxidation, Color, and Volatiles of Raw and Cooked Chicken Breast Meat during Storage

    Directory of Open Access Journals (Sweden)

    Marwan Al-Hijazeen

    2016-06-01

    Full Text Available The objective of this study was to determine the effect of tannic acid (TA on the oxidative stability and the quality characteristics of ground chicken breast meat. Five treatments including (1 control (none added, (2 2.5 ppm TA, (3 5 ppm TA, (4 10 ppm TA, and (5 5 ppm butylated hydroxyanisole (BHA were added to boneless, skinless ground chicken breast meat, and used for both raw and cooked meat studies. For the raw meat study, the ground chicken breast meat was packaged in oxygen-permeable bags and stored at 4 °C for 7 days. For the cooked study, raw ground meat samples were vacuum-packaged in oxygen-impermeable vacuum bags, cooked in-bag to the internal temperature of 75 °C, re-packaged in oxygen-permeable bags, and then stored. Both raw and cooked meats were analyzed for lipid and protein oxidation, color, and volatiles (cooked meat only at 0, 3, and 7 days of storage. Raw meats with 10 ppm of TA added had significantly (p ≤ 0.05 lower lipid and protein oxidation than other treatments during storage. In addition, TA at 10 ppm level maintained the highest color a*- and L*-values during storage. Cooked chicken breast meat with 5 and 10 ppm TA added produced significantly (p ≤ 0.05 lower amounts of off-odor volatiles than other treatments. Among the volatile compounds, the amount of hexanal increased rapidly during storage for cooked meat. However, meats with 5 and 10 ppm TA added showed the lowest amount of hexanal and other aldehydes related to lipid oxidation, indicating a strong antioxidant effect of TA in cooked chicken breast meat. Furthermore, the differences in aldehydes among the treatments were bigger in cooked than in raw meat, indicating that the antioxidant effect of TA in cooked meat was greater than that in raw meat. Therefore, TA at >5 ppm can be used as a good natural preservative in cooked chicken meat to maintain its quality during storage.

  6. Proteome approaches to characterize seed storage proteins related to ditelocentric chromosomes in common wheat (Triticum aestivum L.).

    Science.gov (United States)

    Islam, Nazrul; Woo, Sun-Hee; Tsujimoto, Hisashi; Kawasaki, Hiroshi; Hirano, Hisashi

    2002-09-01

    Changes in protein composition of wheat endosperm proteome were investigated in 39 ditelocentric chromosome lines of common wheat (Triticum aestivum L.) cv. Chinese Spring. Two-dimensional gel electrophoresis followed by Coomassie Brilliant Blue staining has resolved a total of 105 protein spots in a gel. Quantitative image analysis of protein spots was performed by PDQuest. Variations in protein spots between the euploid and the 39 ditelocentric lines were evaluated by spot number, appearance, disappearance and intensity. A specific spot present in all gels was taken as an internal standard, and the intensity of all other spots was calculated as the ratio of the internal standard. Out of the 1755 major spots detected in 39 ditelocentric lines, 1372 (78%) spots were found variable in different spot parameters: 147 (11%) disappeared, 978 (71%) up-regulated and 247 (18%) down-regulated. Correlation studies in changes in protein intensities among 24 protein spots across the ditelocentric lines were performed. High correlations in changes of protein intensities were observed among the proteins encoded by genes located in the homoeologous arms. Locations of structural genes controlling 26 spots were identified in 10 chromosomal arms. Multiple regulators of the same protein located at various chromosomal arms were also noticed. Identification of structural genes for most of the proteins was found difficult due to multiple regulators encoding the same protein. Two novel subunits (1B(Z,) 1BDz), the structure of which are very similar to the high molecular weight glutenin subunit 12, were identified, and the chromosome arm locations of these subunits were assigned.

  7. Quality of black beans as a function of long-term storage and moldy development: Chemical and functional properties of flour and isolated protein.

    Science.gov (United States)

    Ferreira, Cristiano Dietrich; Ziegler, Valmor; Lindemann, Igor da Silva; Hoffmann, Jessica Fernanda; Vanier, Nathan Levien; Oliveira, Maurício de

    2018-04-25

    The aim of this study was to evaluate the effects of moisture content and storage temperature on the percentage of moldy and fermented beans, mycotoxins levels, phenolic acids content, pasting properties of whole flour, as well as functional and thermal properties of protein isolates from black beans stored for 12 months. Beans stored under 14%/32 °C exhibited 16% of fermented grains, while at 17%/25 °C (42.3%) and 17%/32 °C (93.5%) of moldy plus fermented grains, named drastic conditions (DC). Mycotoxins were not present in grains from all storage conditions. Reduction of gallic, caffeic, and p-hydroxybenzoic acid contents, and increase of sinapic acid were observed in DC. Reduction of peak, final, and setback viscosities of bean flours in DC indicate the application in refrigerated and frozen products. The increase in foaming and reduction in foam degradation of the proteins highlights their use in beverages where the foam is an important factor. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Effects of immobilization and aerobic training on proteins related to intramuscular substrate storage and metabolism in young and older men

    DEFF Research Database (Denmark)

    Vigelsø Hansen, Andreas; Gram, Martin; Wiuff, Caroline

    2016-01-01

    by aerobic training in young and older men. METHODS: 17 young (23 ± 1 years, 24 ± 1 kg m(-2), and 20 ± 2% body fat) and 15 older men (68 ± 1 years; 27 ± 1 kg m(-2), and 29 ± 2% body fat) underwent 2 weeks' one leg immobilization followed by 6 weeks' cycle training. Biopsies were obtained from m. vastus...... lateralis just before immobilization (at inclusion), after immobilization, and the after 6 weeks' training. The biopsies were analyzed for muscle substrates; muscle perilipin protein (PLIN), glycogen synthase (GS), synaptosomal-associated protein of 23 kDa (SNAP23) protein content, and muscle 3-hydroxyacyl...... GS (74%) protein compared to the older men. Immobilization decreased and training restored HAD activity, GS and SNAP23 protein content in young and older men. CONCLUSION: Evidence of age-related metabolic inflexibility is presented, seen as body fat and IMTG accumulation. The question arises...

  9. Spreading of a granular droplet

    Science.gov (United States)

    Clement, Eric; Sanchez, Ivan; Raynaud, Franck; Lanuza, Jose; Andreotti, Bruno; Aranson, Igor

    2008-03-01

    The influence of controlled vibrations on the granular rheology is investigated in a specifically designed experiment in which a granular film spreads under the action of horizontal vibrations. A nonlinear diffusion equation is derived theoretically that describes the evolution of the deposit shape. A self-similar parabolic shape (the``granular droplet'') and a spreading dynamics are predicted that both agree quantitatively with the experimental results. The theoretical analysis is used to extract effective friction coefficients between the base and the granular layer under sustained and controlled vibrations. A shear thickening regime characteristic of dense granular flows is evidenced at low vibration energy, both for glass beads and natural sand. Conversely, shear thinning is observed at high agitation.

  10. Models of lipid droplets growth and fission in adipocyte cells

    Energy Technology Data Exchange (ETDEWEB)

    Boschi, Federico, E-mail: federico.boschi@univr.it [Department of Computer Science, University of Verona, Strada Le Grazie 15, 37134 Verona (Italy); Rizzatti, Vanni; Zamboni, Mauro [Department of Medicine, Geriatric Section, University of Verona, Piazzale Stefani 1, 37126 Verona (Italy); Sbarbati, Andrea [Department of Neurological and Movement Sciences, University of Verona, Strada Le Grazie 8, 37134 Verona (Italy)

    2015-08-15

    Lipid droplets (LD) are spherical cellular inclusion devoted to lipids storage. It is well known that excessive accumulation of lipids leads to several human worldwide diseases like obesity, type 2 diabetes, hepatic steatosis and atherosclerosis. LDs' size range from fraction to one hundred of micrometers in adipocytes and is related to the lipid content, but their growth is still a puzzling question. It has been suggested that LDs can grow in size due to the fusion process by which a larger LD is obtained by the merging of two smaller LDs, but these events seems to be rare and difficult to be observed. Many other processes are thought to be involved in the number and growth of LDs, like the de novo formation and the growth through additional neutral lipid deposition in pre-existing droplets. Moreover the number and size of LDs are influenced by the catabolism and the absorption or interaction with other organelles. The comprehension of these processes could help in the confinement of the pathologies related to lipid accumulation. In this study the LDs' size distribution, number and the total volume of immature (n=12), mature (n=12, 10-days differentiated) and lipolytic (n=12) 3T3-L1 adipocytes were considered. More than 11,000 LDs were measured in the 36 cells after Oil Red O staining. In a previous work Monte Carlo simulations were used to mimic the fusion process alone between LDs. We found that, considering the fusion as the only process acting on the LDs, the size distribution in mature adipocytes can be obtained with numerical simulation starting from the size distribution in immature cells provided a very high rate of fusion events. In this paper Monte Carlo simulations were developed to mimic the interaction between LDs taking into account many other processes in addition to fusion (de novo formation and the growth through additional neutral lipid deposition in pre-existing droplets) in order to reproduce the LDs growth and we also simulated the

  11. Models of lipid droplets growth and fission in adipocyte cells

    International Nuclear Information System (INIS)

    Boschi, Federico; Rizzatti, Vanni; Zamboni, Mauro; Sbarbati, Andrea

    2015-01-01

    Lipid droplets (LD) are spherical cellular inclusion devoted to lipids storage. It is well known that excessive accumulation of lipids leads to several human worldwide diseases like obesity, type 2 diabetes, hepatic steatosis and atherosclerosis. LDs' size range from fraction to one hundred of micrometers in adipocytes and is related to the lipid content, but their growth is still a puzzling question. It has been suggested that LDs can grow in size due to the fusion process by which a larger LD is obtained by the merging of two smaller LDs, but these events seems to be rare and difficult to be observed. Many other processes are thought to be involved in the number and growth of LDs, like the de novo formation and the growth through additional neutral lipid deposition in pre-existing droplets. Moreover the number and size of LDs are influenced by the catabolism and the absorption or interaction with other organelles. The comprehension of these processes could help in the confinement of the pathologies related to lipid accumulation. In this study the LDs' size distribution, number and the total volume of immature (n=12), mature (n=12, 10-days differentiated) and lipolytic (n=12) 3T3-L1 adipocytes were considered. More than 11,000 LDs were measured in the 36 cells after Oil Red O staining. In a previous work Monte Carlo simulations were used to mimic the fusion process alone between LDs. We found that, considering the fusion as the only process acting on the LDs, the size distribution in mature adipocytes can be obtained with numerical simulation starting from the size distribution in immature cells provided a very high rate of fusion events. In this paper Monte Carlo simulations were developed to mimic the interaction between LDs taking into account many other processes in addition to fusion (de novo formation and the growth through additional neutral lipid deposition in pre-existing droplets) in order to reproduce the LDs growth and we also simulated the

  12. A Conserved Role for Atlastin GTPases in Regulating Lipid Droplet Size

    Directory of Open Access Journals (Sweden)

    Robin W. Klemm

    2013-05-01

    Full Text Available Lipid droplets (LDs are the major fat storage organelles in eukaryotic cells, but how their size is regulated is unknown. Using genetic screens in C. elegans for LD morphology defects in intestinal cells, we found that mutations in atlastin, a GTPase required for homotypic fusion of endoplasmic reticulum (ER membranes, cause not only ER morphology defects, but also a reduction in LD size. Similar results were obtained after depletion of atlastin or expression of a dominant-negative mutant, whereas overexpression of atlastin had the opposite effect. Atlastin depletion in Drosophila fat bodies also reduced LD size and decreased triglycerides in whole animals, sensitizing them to starvation. In mammalian cells, co-overexpression of atlastin-1 and REEP1, a paralog of the ER tubule-shaping protein DP1/REEP5, generates large LDs. The effect of atlastin-1 on LD size correlates with its activity to promote membrane fusion in vitro. Our results indicate that atlastin-mediated fusion of ER membranes is important for LD size regulation.

  13. Experimental and computational analysis of a large protein network that controls fat storage reveals the design principles of a signaling network.

    Directory of Open Access Journals (Sweden)

    Bader Al-Anzi

    2015-05-01

    Full Text Available An approach combining genetic, proteomic, computational, and physiological analysis was used to define a protein network that regulates fat storage in budding yeast (Saccharomyces cerevisiae. A computational analysis of this network shows that it is not scale-free, and is best approximated by the Watts-Strogatz model, which generates "small-world" networks with high clustering and short path lengths. The network is also modular, containing energy level sensing proteins that connect to four output processes: autophagy, fatty acid synthesis, mRNA processing, and MAP kinase signaling. The importance of each protein to network function is dependent on its Katz centrality score, which is related both to the protein's position within a module and to the module's relationship to the network as a whole. The network is also divisible into subnetworks that span modular boundaries and regulate different aspects of fat metabolism. We used a combination of genetics and pharmacology to simultaneously block output from multiple network nodes. The phenotypic results of this blockage define patterns of communication among distant network nodes, and these patterns are consistent with the Watts-Strogatz model.

  14. Experimental and computational analysis of a large protein network that controls fat storage reveals the design principles of a signaling network.

    Science.gov (United States)

    Al-Anzi, Bader; Arpp, Patrick; Gerges, Sherif; Ormerod, Christopher; Olsman, Noah; Zinn, Kai

    2015-05-01

    An approach combining genetic, proteomic, computational, and physiological analysis was used to define a protein network that regulates fat storage in budding yeast (Saccharomyces cerevisiae). A computational analysis of this network shows that it is not scale-free, and is best approximated by the Watts-Strogatz model, which generates "small-world" networks with high clustering and short path lengths. The network is also modular, containing energy level sensing proteins that connect to four output processes: autophagy, fatty acid synthesis, mRNA processing, and MAP kinase signaling. The importance of each protein to network function is dependent on its Katz centrality score, which is related both to the protein's position within a module and to the module's relationship to the network as a whole. The network is also divisible into subnetworks that span modular boundaries and regulate different aspects of fat metabolism. We used a combination of genetics and pharmacology to simultaneously block output from multiple network nodes. The phenotypic results of this blockage define patterns of communication among distant network nodes, and these patterns are consistent with the Watts-Strogatz model.

  15. Internal flow inside droplets within a concentrated emulsion during droplet rearrangement

    Science.gov (United States)

    Leong, Chia Min; Gai, Ya; Tang, Sindy K. Y.

    2018-03-01

    Droplet microfluidics, in which each droplet serves as a micro-reactor, has found widespread use in high-throughput biochemical screening applications. These droplets are often concentrated at various steps to form a concentrated emulsion. As part of a serial interrogation and sorting process, such concentrated emulsions are typically injected into a tapered channel leading to a constriction that fits one drop at a time for the probing of droplet content in a serial manner. The flow physics inside the droplets under these flow conditions are not well understood but are critical for predicting and controlling the mixing of reagents inside the droplets as reactors. Here we investigate the flow field inside droplets of a concentrated emulsion flowing through a tapered microchannel using micro-particle image velocimetry. The confining geometry of the channel forces the number of rows of drops to reduce by one at specific and uniformly spaced streamwise locations, which are referred to as droplet rearrangement zones. Within each rearrangement zone, the phase-averaged velocity results show that the motion of the droplets involved in the rearrangement process, also known as a T1 event, creates vortical structures inside themselves and their adjacent droplets. These flow structures increase the circulation inside droplets up to 2.5 times the circulation in droplets at the constriction. The structures weaken outside of the rearrangement zones suggesting that the flow patterns created by the T1 process are transient. The time scale of circulation is approximately the same as the time scale of a T1 event. Outside of the rearrangement zones, flow patterns in the droplets are determined by the relative velocity between the continuous and disperse phases.

  16. Spontaneous droplet trampolining on rigid superhydrophobic surfaces

    Science.gov (United States)

    Schutzius, Thomas M.; Jung, Stefan; Maitra, Tanmoy; Graeber, Gustav; Köhme, Moritz; Poulikakos, Dimos

    2015-11-01

    Spontaneous removal of condensed matter from surfaces is exploited in nature and in a broad range of technologies to achieve self-cleaning, anti-icing and condensation control. But despite much progress, our understanding of the phenomena leading to such behaviour remains incomplete, which makes it challenging to rationally design surfaces that benefit from its manifestation. Here we show that water droplets resting on superhydrophobic textured surfaces in a low-pressure environment can self-remove through sudden spontaneous levitation and subsequent trampoline-like bouncing behaviour, in which sequential collisions with the surface accelerate the droplets. These collisions have restitution coefficients (ratios of relative speeds after and before collision) greater than unity despite complete rigidity of the surface, and thus seemingly violate the second law of thermodynamics. However, these restitution coefficients result from an overpressure beneath the droplet produced by fast droplet vaporization while substrate adhesion and surface texture restrict vapour flow. We also show that the high vaporization rates experienced by the droplets and the associated cooling can result in freezing from a supercooled state that triggers a sudden increase in vaporization, which in turn boosts the levitation process. This effect can spontaneously remove surface icing by lifting away icy drops the moment they freeze. Although these observations are relevant only to systems in a low-pressure environment, they show how surface texturing can produce droplet-surface interactions that prohibit liquid and freezing water-droplet retention on surfaces.

  17. Mass spectrometry of acoustically levitated droplets.

    Science.gov (United States)

    Westphall, Michael S; Jorabchi, Kaveh; Smith, Lloyd M

    2008-08-01

    Containerless sample handling techniques such as acoustic levitation offer potential advantages for mass spectrometry, by eliminating surfaces where undesired adsorption/desorption processes can occur. In addition, they provide a unique opportunity to study fundamental aspects of the ionization process as well as phenomena occurring at the air-droplet interface. Realizing these advantages is contingent, however, upon being able to effectively interface levitated droplets with a mass spectrometer, a challenging task that is addressed in this report. We have employed a newly developed charge and matrix-assisted laser desorption/ionization (CALDI) technique to obtain mass spectra from a 5-microL acoustically levitated droplet containing peptides and an ionic matrix. A four-ring electrostatic lens is used in conjunction with a corona needle to produce bursts of corona ions and to direct those ions toward the droplet, resulting in droplet charging. Analyte ions are produced from the droplet by a 337-nm laser pulse and detected by an atmospheric sampling mass spectrometer. The ion generation and extraction cycle is repeated at 20 Hz, the maximum operating frequency of the laser employed. It is shown in delayed ion extraction experiments that both positive and negative ions are produced, behavior similar to that observed for atmospheric pressure matrix-assisted laser absorption/ionization. No ion signal is observed in the absence of droplet charging. It is likely, although not yet proven, that the role of the droplet charging is to increase the strength of the electric field at the surface of the droplet, reducing charge recombination after ion desorption.

  18. Effects of immobilization and aerobic training on proteins related to intramuscular substrate storage and metabolism in young and older men.

    Science.gov (United States)

    Vigelsø, Andreas; Gram, Martin; Wiuff, Caroline; Hansen, Christina Neigaard; Prats, Clara; Dela, Flemming; Helge, Jørn Wulff

    2016-03-01

    Aging and inactivity lead to skeletal muscle metabolic inflexibility, but the underlying molecular mechanisms are not entirely elucidated. Therefore, we investigated how muscle lipid and glycogen stores and major regulatory proteins were affected by short-term immobilization followed by aerobic training in young and older men. 17 young (23 ± 1 years, 24 ± 1 kg m(-2), and 20 ± 2% body fat) and 15 older men (68 ± 1 years; 27 ± 1 kg m(-2), and 29 ± 2% body fat) underwent 2 weeks' one leg immobilization followed by 6 weeks' cycle training. Biopsies were obtained from m. vastus lateralis just before immobilization (at inclusion), after immobilization, and the after 6 weeks' training. The biopsies were analyzed for muscle substrates; muscle perilipin protein (PLIN), glycogen synthase (GS), synaptosomal-associated protein of 23 kDa (SNAP23) protein content, and muscle 3-hydroxyacyl-CoA dehydrogenase (HAD) activity The older men had higher intramuscular triglyceride (IMTG) (73 %) and Glycogen (16%) levels compared to the young men, and IMTG tended to increase with immobilization. PLIN2 and 3 protein content increased with immobilization in the older men only. The young men had higher GS (74%) protein compared to the older men. Immobilization decreased and training restored HAD activity, GS and SNAP23 protein content in young and older men. Evidence of age-related metabolic inflexibility is presented, seen as body fat and IMTG accumulation. The question arises as to whether IMTG accumulation in the older men is caused by or leading to the increase in PLIN2 and 3 protein content. Training decreased body fat and IMTG levels in both young and older men; hence, training should be prioritized to reduce the detrimental effect of aging on metabolism.

  19. Lipid Droplet Formation Is Dispensable for Endoplasmic Reticulum-associated Degradation*

    Science.gov (United States)

    Olzmann, James A.; Kopito, Ron R.

    2011-01-01

    Proteins that fail to fold or assemble in the endoplasmic reticulum (ER) are destroyed by cytoplasmic proteasomes through a process known as ER-associated degradation. Substrates of this pathway are initially sequestered within the ER lumen and must therefore be dislocated across the ER membrane to be degraded. It has been proposed that generation of bicellar structures during lipid droplet formation may provide an “escape hatch” through which misfolded proteins, toxins, and viruses can exit the ER. We have directly tested this hypothesis by exploiting yeast strains defective in lipid droplet formation. Our data demonstrate that lipid droplet formation is dispensable for the dislocation of a plant toxin and the degradation of both soluble and integral membrane glycoproteins. PMID:21693705

  20. Foam droplet separation for nanoparticle synthesis

    International Nuclear Information System (INIS)

    Tyree, Corey A.; Allen, Jonathan O.

    2008-01-01

    A novel approach to nanoparticle synthesis was developed whereby foam bubble bursting produced aerosol droplets, an approach patterned after the marine foam aerosol cycle. The droplets were dried to remove solvent, leaving nanometer-sized particles composed of precursor material. Nanoparticles composed of sodium chloride (mean diameter, D-bar p ∼ 100 nm), phosphotungstic acid (D-bar p ∼ 55 nm), and bovine insulin (D p ∼ 5-30 nm) were synthesized. Foam droplet separation can be carried out at ambient temperature and pressure. The 'soft' nature of the process makes it compatible with a wide range of materials

  1. Diffusion and evaporation of a liquid droplet

    Science.gov (United States)

    Shukla, K. N.

    1980-06-01

    The process of evaporation and diffusion of a spherical liquid droplet in an atmosphere of noncondensable gas is studied theoretically. An equation for the shrinkage of the radius of the droplet is derived on the basis of continuity and momentum equations. Further, a conjugate problem consisting of the energy and mass balance for the gaseous environment is formulated. An approximation of thin thermal and diffusion boundary-layers is introduced to simplify the analysis. Results are presented for methanol-nitrogen, ammonia-nitrogen, and sodium-argon systems. It has been observed that the droplet of highly viscous fluid exhibits rapid contraction.

  2. Electronically droplet energy harvesting using piezoelectric cantilevers

    KAUST Repository

    Al Ahmad, Mahmoud Al

    2012-01-01

    A report is presented on free falling droplet energy harvesting using piezoelectric cantilevers. The harvester incorporates a multimorph clamped-free cantilever which is composed of five layers of lead zirconate titanate piezoelectric thick films. During the impact, the droplet kinetic energy is transferred into the form of mechanical stress forcing the piezoelectric structure to vibrate. Experimental results show energy of 0.3 μJ per droplet. The scenario of moderate falling drop intensity, i.e. 230 drops per second, yields a total energy of 400 μJ. © 2012 The Institution of Engineering and Technology.

  3. Heat exchanges between droplets and atmosphere

    International Nuclear Information System (INIS)

    Yadigaroglu, Georges.

    1975-01-01

    Data necessary for calculating the droplet cooling in wet cooling systems are surveyed. This cooling obeys the laws of simultaneous heat and mass transfer. Exchanges with a solid sphere moving inside a surrounding fluid medium are first examined. The corrections needed for taking into account various secondary effects (circulation in the droplet, lack of sphericity, oscillations, etc...) are then dealt with. Some data necessary for calculating the trajectories of the droplets and their behavior in a cooling system are included (diameter distribution, limit velocities, decay thresholds, etc...). Finally, calculation methods applying to spray systems, as well as wet towers broadly outlined [fr

  4. Printed droplet microfluidics for on demand dispensing of picoliter droplets and cells.

    Science.gov (United States)

    Cole, Russell H; Tang, Shi-Yang; Siltanen, Christian A; Shahi, Payam; Zhang, Jesse Q; Poust, Sean; Gartner, Zev J; Abate, Adam R

    2017-08-15

    Although the elementary unit of biology is the cell, high-throughput methods for the microscale manipulation of cells and reagents are limited. The existing options either are slow, lack single-cell specificity, or use fluid volumes out of scale with those of cells. Here we present printed droplet microfluidics, a technology to dispense picoliter droplets and cells with deterministic control. The core technology is a fluorescence-activated droplet sorter coupled to a specialized substrate that together act as a picoliter droplet and single-cell printer, enabling high-throughput generation of intricate arrays of droplets, cells, and microparticles. Printed droplet microfluidics provides a programmable and robust technology to construct arrays of defined cell and reagent combinations and to integrate multiple measurement modalities together in a single assay.

  5. Modest hypoxia significantly reduces triglyceride content and lipid droplet size in 3T3-L1 adipocytes

    Energy Technology Data Exchange (ETDEWEB)

    Hashimoto, Takeshi, E-mail: thashimo@fc.ritsumei.ac.jp [Faculty of Sport and Health Science, Ritsumeikan University, 1-1-1 Nojihigashi, Kusatsu, Shiga 525-8577 (Japan); Yokokawa, Takumi; Endo, Yuriko [Faculty of Sport and Health Science, Ritsumeikan University, 1-1-1 Nojihigashi, Kusatsu, Shiga 525-8577 (Japan); Iwanaka, Nobumasa [Ritsumeikan Global Innovation Research Organization, Ritsumeikan University, 1-1-1 Nojihigashi, Kusatsu, Shiga 525-8577 (Japan); Higashida, Kazuhiko [Faculty of Sport and Health Science, Ritsumeikan University, 1-1-1 Nojihigashi, Kusatsu, Shiga 525-8577 (Japan); Faculty of Sport Science, Waseda University, 2-579-15 Mikajima, Tokorozawa, Saitama 359-1192 (Japan); Taguchi, Sadayoshi [Faculty of Sport and Health Science, Ritsumeikan University, 1-1-1 Nojihigashi, Kusatsu, Shiga 525-8577 (Japan)

    2013-10-11

    Highlights: •Long-term hypoxia decreased the size of LDs and lipid storage in 3T3-L1 adipocytes. •Long-term hypoxia increased basal lipolysis in 3T3-L1 adipocytes. •Hypoxia decreased lipid-associated proteins in 3T3-L1 adipocytes. •Hypoxia decreased basal glucose uptake and lipogenic proteins in 3T3-L1 adipocytes. •Hypoxia-mediated lipogenesis may be an attractive therapeutic target against obesity. -- Abstract: Background: A previous study has demonstrated that endurance training under hypoxia results in a greater reduction in body fat mass compared to exercise under normoxia. However, the cellular and molecular mechanisms that underlie this hypoxia-mediated reduction in fat mass remain uncertain. Here, we examine the effects of modest hypoxia on adipocyte function. Methods: Differentiated 3T3-L1 adipocytes were incubated at 5% O{sub 2} for 1 week (long-term hypoxia, HL) or one day (short-term hypoxia, HS) and compared with a normoxia control (NC). Results: HL, but not HS, resulted in a significant reduction in lipid droplet size and triglyceride content (by 50%) compared to NC (p < 0.01). As estimated by glycerol release, isoproterenol-induced lipolysis was significantly lowered by hypoxia, whereas the release of free fatty acids under the basal condition was prominently enhanced with HL compared to NC or HS (p < 0.01). Lipolysis-associated proteins, such as perilipin 1 and hormone-sensitive lipase, were unchanged, whereas adipose triglyceride lipase and its activator protein CGI-58 were decreased with HL in comparison to NC. Interestingly, such lipogenic proteins as fatty acid synthase, lipin-1, and peroxisome proliferator-activated receptor gamma were decreased. Furthermore, the uptake of glucose, the major precursor of 3-glycerol phosphate for triglyceride synthesis, was significantly reduced in HL compared to NC or HS (p < 0.01). Conclusion: We conclude that hypoxia has a direct impact on reducing the triglyceride content and lipid droplet size via

  6. Modest hypoxia significantly reduces triglyceride content and lipid droplet size in 3T3-L1 adipocytes

    International Nuclear Information System (INIS)

    Hashimoto, Takeshi; Yokokawa, Takumi; Endo, Yuriko; Iwanaka, Nobumasa; Higashida, Kazuhiko; Taguchi, Sadayoshi

    2013-01-01

    Highlights: •Long-term hypoxia decreased the size of LDs and lipid storage in 3T3-L1 adipocytes. •Long-term hypoxia increased basal lipolysis in 3T3-L1 adipocytes. •Hypoxia decreased lipid-associated proteins in 3T3-L1 adipocytes. •Hypoxia decreased basal glucose uptake and lipogenic proteins in 3T3-L1 adipocytes. •Hypoxia-mediated lipogenesis may be an attractive therapeutic target against obesity. -- Abstract: Background: A previous study has demonstrated that endurance training under hypoxia results in a greater reduction in body fat mass compared to exercise under normoxia. However, the cellular and molecular mechanisms that underlie this hypoxia-mediated reduction in fat mass remain uncertain. Here, we examine the effects of modest hypoxia on adipocyte function. Methods: Differentiated 3T3-L1 adipocytes were incubated at 5% O 2 for 1 week (long-term hypoxia, HL) or one day (short-term hypoxia, HS) and compared with a normoxia control (NC). Results: HL, but not HS, resulted in a significant reduction in lipid droplet size and triglyceride content (by 50%) compared to NC (p < 0.01). As estimated by glycerol release, isoproterenol-induced lipolysis was significantly lowered by hypoxia, whereas the release of free fatty acids under the basal condition was prominently enhanced with HL compared to NC or HS (p < 0.01). Lipolysis-associated proteins, such as perilipin 1 and hormone-sensitive lipase, were unchanged, whereas adipose triglyceride lipase and its activator protein CGI-58 were decreased with HL in comparison to NC. Interestingly, such lipogenic proteins as fatty acid synthase, lipin-1, and peroxisome proliferator-activated receptor gamma were decreased. Furthermore, the uptake of glucose, the major precursor of 3-glycerol phosphate for triglyceride synthesis, was significantly reduced in HL compared to NC or HS (p < 0.01). Conclusion: We conclude that hypoxia has a direct impact on reducing the triglyceride content and lipid droplet size via

  7. Increased serum C-reactive protein level is associated with increased storage lower urinary tract symptoms in men with benign prostatic hyperplasia.

    Directory of Open Access Journals (Sweden)

    Shun-Fa Hung

    Full Text Available OBJECTIVE: Chronic inflammation is considered as one of the contributing mechanisms of lower urinary tract symptoms (LUTS. Serum C-reactive protein (CRP level is the widely used biomarker of inflammatory status. This study investigated the association between serum CRP level in men with benign prostatic hyperplasia (BPH and lower urinary tract symptoms (LUTS before and after medical treatment. METHODS: A total of 853 men with BPH and LUTS were enrolled. All patients completed the International Prostate Symptoms Score (IPSS questionnaire and urological examinations. The parameters of uroflowmetry (maximum flow rate, Qmax; voided volume, VV, post-void residual (PVR, total prostate volume (TPV and transition zone index (TZI, serum prostate specific antigen (PSA, and serum CRP levels were obtained. All patients were treated with alpha-blocker or antimuscarinic agent based on the IPSS voiding to storage subscore ratio (IPSS-V/S. Correlation analyses were performed between serum CRP levels with age, IPSS, TPV, TZI, Qmax, PVR, VV, PSA and between baseline and post treatment. RESULTS: The mean age was 66.9 ± 11.6 years old and the mean serum CRP levels were 0.31 ± 0.43 mg/dL. Univariate analyses revealed serum CRP levels were significantly associated with age (p<0.001, PSA levels (p = 0.005 and VV (p = 0.017, but not significantly associated with TPV (p = 0.854 or PVR (p = 0.068. CRP levels were positively associated with urgency (p<0.001 and nocturia (p<0.001 subscore of IPSS, total IPSS (p = 0.008 and storage IPSS (p<0.001 and negatively associated with IPSS- V/S ratio (p = 0.014. Multivariate analyses revealed that serum CRP levels were significantly associated with age (p = 0.004 and storage IPSS subscore p<0.001. Patients with IPSS-V/S<1 and treated with tolterodine for 3 months had significant decrease of CRP levels after treatment. CONCLUSION: Serum CRP levels are associated with storage LUTS and sensory bladder disorders, suggesting chronic

  8. Janus droplets: liquid marbles coated with dielectric/semiconductor particles.

    Science.gov (United States)

    Bormashenko, Edward; Bormashenko, Yelena; Pogreb, Roman; Gendelman, Oleg

    2011-01-04

    The manufacturing of water droplets wrapped with two different powders, carbon black (semiconductor) and polytetrafluoroethylene (dielectric), is presented. Droplets composed of two hemispheres (Janus droplets) characterized by various physical and chemical properties are reported first. Watermelon-like striped liquid marbles are reported. Janus droplets remained stable on solid and liquid supports and could be activated with an electric field.

  9. Self-propelled oil droplets consuming "fuel" surfactant

    DEFF Research Database (Denmark)

    Toyota, Taro; Maru, Naoto; Hanczyc, Martin M

    2009-01-01

    A micrometer-sized oil droplet of 4-octylaniline containing 5 mol % of an amphiphilic catalyst exhibited a self-propelled motion, producing tiny oil droplets, in an aqueous dispersion of an amphiphilic precursor of 4-octylaniline. The tiny droplets on the surface of the self-propelled droplet wer...

  10. Electrostatic charging and control of droplets in microfluidic devices.

    Science.gov (United States)

    Zhou, Hongbo; Yao, Shuhuai

    2013-03-07

    Precharged droplets can facilitate manipulation and control of low-volume liquids in droplet-based microfluidics. In this paper, we demonstrate non-contact electrostatic charging of droplets by polarizing a neutral droplet and splitting it into two oppositely charged daughter droplets in a T-junction microchannel. We performed numerical simulation to analyze the non-contact charging process and proposed a new design with a notch at the T-junction in aid of droplet splitting for more efficient charging. We experimentally characterized the induced charge in droplets in microfabricated devices. The experimental results agreed well with the simulation. Finally, we demonstrated highly effective droplet manipulation in a path selection unit appending to the droplet charging. We expect our work could enable precision manipulation of droplets for more complex liquid handling in microfluidics and promote electric-force based manipulation in 'lab-on-a-chip' systems.

  11. Effects of droplet interactions on droplet transport at intermediate Reynolds numbers

    Science.gov (United States)

    Shuen, Jian-Shun

    1987-01-01

    Effects of droplet interactions on drag, evaporation, and combustion of a planar droplet array, oriented perpendicular to the approaching flow, are studied numerically. The three-dimensional Navier-Stokes equations, with variable thermophysical properties, are solved using finite-difference techniques. Parameters investigated include the droplet spacing, droplet Reynolds number, approaching stream oxygen concentration, and fuel type. Results are obtained for the Reynolds number range of 5 to 100, droplet spacings from 2 to 24 diameters, oxygen concentrations of 0.1 and 0.2, and methanol and n-butanol fuels. The calculations show that the gasification rates of interacting droplets decrease as the droplet spacings decrease. The reduction in gasification rates is significant only at small spacings and low Reynolds numbers. For the present array orientation, the effects of interactions on the gasification rates diminish rapidly for Reynolds numbers greater than 10 and spacings greater than 6 droplet diameters. The effects of adjacent droplets on drag are shown to be small.

  12. Multi-scale simulation of droplet-droplet interactions and coalescence

    CSIR Research Space (South Africa)

    Musehane, Ndivhuwo M

    2016-10-01

    Full Text Available Conference on Computational and Applied Mechanics Potchefstroom 3–5 October 2016 Multi-scale simulation of droplet-droplet interactions and coalescence 1,2Ndivhuwo M. Musehane?, 1Oliver F. Oxtoby and 2Daya B. Reddy 1. Aeronautic Systems, Council... topology changes that result when droplets interact. This work endeavours to eliminate the need to use empirical correlations based on phenomenological models by developing a multi-scale model that predicts the outcome of a collision between droplets from...

  13. Substrate curvature gradient drives rapid droplet motion.

    Science.gov (United States)

    Lv, Cunjing; Chen, Chao; Chuang, Yin-Chuan; Tseng, Fan-Gang; Yin, Yajun; Grey, Francois; Zheng, Quanshui

    2014-07-11

    Making small liquid droplets move spontaneously on solid surfaces is a key challenge in lab-on-chip and heat exchanger technologies. Here, we report that a substrate curvature gradient can accelerate micro- and nanodroplets to high speeds on both hydrophilic and hydrophobic substrates. Experiments for microscale water droplets on tapered surfaces show a maximum speed of 0.42  m/s, 2 orders of magnitude higher than with a wettability gradient. We show that the total free energy and driving force exerted on a droplet are determined by the substrate curvature and substrate curvature gradient, respectively. Using molecular dynamics simulations, we predict nanoscale droplets moving spontaneously at over 100  m/s on tapered surfaces.

  14. Strange particle production from quark matter droplets

    International Nuclear Information System (INIS)

    Werner, K.; Hladik, M.

    1995-01-01

    We recently introduced new methods to study ultrarelativistic nuclear scattering by providing a link between the string model approach and a thermal description. The string model is used to provide information about fluctuations in energy density. Regions of high energy density are considered to be quark matter droplets and treated macroscopically. At SPS energies, we find mainly medium size droplets---with energies up to few tens of Gev. A key issue is the microcanonical treatment of individual quark matter droplets. Each droplet hadronizes instantaneously according to the available n-body phase space. Due to the huge number of possible hadron configurations, special Monte Carlo techniques have been developed to calculate this disintegration. We present results concerning the production of strange particles from such a hadronization as compared to string decay. copyright 1995 American Institute of Physics

  15. Manipulation of microfluidic droplets by electrorheological fluid

    KAUST Repository

    Zhang, Menying; Gong, Xiuqing; Wen, Weijia

    2009-01-01

    Microfluidics, especially droplet microfluidics, attracts more and more researchers from diverse fields, because it requires fewer materials and less time, produces less waste and has the potential of highly integrated and computer

  16. Manipulation of microfluidic droplets by electrorheological fluid

    KAUST Repository

    Zhang, Menying

    2009-09-01

    Microfluidics, especially droplet microfluidics, attracts more and more researchers from diverse fields, because it requires fewer materials and less time, produces less waste and has the potential of highly integrated and computer-controlled reaction processes for chemistry and biology. Electrorheological fluid, especially giant electrorheological fluid (GERF), which is considered as a kind of smart material, has been applied to the microfluidic systems to achieve active and precise control of fluid by electrical signal. In this review article, we will introduce recent results of microfluidic droplet manipulation, GERF and some pertinent achievements by introducing GERF into microfluidic system: digital generation, manipulation of "smart droplets" and droplet manipulation by GERF. Once it is combined with real-time detection, integrated chip with multiple functions can be realized. © 2009 Wiley-VCH Verlag GmbH & Co. KGaA.

  17. Electronically droplet energy harvesting using piezoelectric cantilevers

    KAUST Repository

    Al Ahmad, Mahmoud Al; Jabbour, Ghassan E.

    2012-01-01

    A report is presented on free falling droplet energy harvesting using piezoelectric cantilevers. The harvester incorporates a multimorph clamped-free cantilever which is composed of five layers of lead zirconate titanate piezoelectric thick films

  18. Numerical modeling of a vaporizing multicomponent droplet

    Science.gov (United States)

    Megaridis, C. M.; Sirignano, W. A.

    The fundamental processes governing the energy, mass, and momentum exchange between the liquid and gas phases of vaporizing, multicomponent liquid droplets have been investigated. The axisymmetric configuration under consideration consists of an isolated multicomponent droplet vaporizing in a convective environment. The model considers different volatilities of the liquid components, variable liquid properties due to variation of the species concentrations, and non-Fickian multicomponent gaseous diffusion. The bicomponent droplet model was employed to examine the commonly used assumptions of unity Lewis number in the liquid phase and Fickian gaseous diffusion. It is found that the droplet drag coefficients, the vaporization rates, and the related transfer numbers are not influenced by the above assumptions in a significant way.

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

    Science.gov (United States)

    Wachowska, Urszula; Tańska, Małgorzata; Konopka, Iwona

    2016-06-16

    Modern agriculture relies on an integrated approach, where chemical treatment is reduced to a minimum and replaced by biological control that involves the use of active microorganisms. The effect of the antagonistic yeast-like fungus Aureobasidium pullulans on proteins and bioactive compounds (alkylresorcinols, sterols, tocols and carotenoids) in winter wheat grain and on the colonization of wheat kernels by fungal microbiota, mainly Fusarium spp. pathogens, was investigated. Biological treatment contributed to a slight increase contents of tocols, alkylresorcinols and sterols in grain. At the same time, the variation of wheat grain proteins was low and not significant. Application of A. pullulans enhanced the natural yeast colonization after six months of grain storage and inhibited growth of F. culmorum pathogens penetrating wheat kernel. This study demonstrated that an integrated approach of wheat grain protection with the use of the yeast-like fungus A. pullulans reduced kernel colonization by Fusarium spp. pathogens and increased the content of nutritionally beneficial phytochemicals in wheat grain without a loss of gluten proteins responsible for baking value. Copyright © 2016. Published by Elsevier B.V.

  20. The oviducal protein, heat-shock 70-kDa protein 8, improves the long-term survival of ram spermatozoa during storage at 17°C in a commercial extender.

    Science.gov (United States)

    Lloyd, R E; Fazeli, A; Watson, P F; Holt, W V

    2012-01-01

    Poor fertility rates are often observed when fresh ram semen stored in conventional extenders is used for cervical artificial insemination (AI). Heat-shock 70-kDa protein 8 (HSPA8), found within the oviduct, prolongs boar, ram and bull sperm survival at body temperatures in vitro. Here, we aimed to determine whether supplementing extenders (INRA-96 and RSD-1) with HSPA8 (4 µg mL⁻¹) would improve their performance in maintaining freshly collected ram sperm viability and sperm nuclear DNA integrity during storage over 48 h at 17°C. Sperm function was assessed at 1, 6, 24 and 48h and this experiment was repeated using 25 × 10⁶ and 800 × 10⁶ spermatozoa mL⁻¹. INRA96 supplemented with HSPA8 maintained sperm viability significantly better than INRA96 alone at both sperm concentrations. However, sperm nuclear DNA fragmentation (DF) increased significantly during storage using the higher sperm concentration, irrespective of the extender and the protein treatment used. Increasing levels of sperm nuclear DF over time could explain why poor fertility rates are often observed following cervical AI using stored ram semen. However, further research is required to ascertain whether supplementing the commercially available INRA96 extender with HSPA8 will improve fertility rates following cervical AI using stored ram semen.

  1. Faox enzymes inhibited Maillard reaction development during storage both in protein glucose model system and low lactose UHT milk

    NARCIS (Netherlands)

    Troise, A.D.; Dathan, N.A.; Fiore, A.; Roviello, G.; Fiore, Di A.; Caira, S.; Cuollo, M.; Simone, De G.; Fogliano, V.; Monti, S.M.

    2014-01-01

    Fructosamines, also known asAmadori products, are formed by the condensation of glucose with the amino group of amino acids or proteins. These compounds are precursors of advanced glycation end products (AGEs) that can be formed either endogenously during aging and diabetes, and exogenously in

  2. Maturity and storage influence on the apple (Malus domestica) allergen Mal d 3, a nonspecific lipid transfer protein

    NARCIS (Netherlands)

    Sancho, Ana I.; Foxall, Robert; Rigby, Neil M.; Browne, Thomas; Zuidmeer, Laurian; van Ree, Ronald; Waldron, Keith W.; Mills, E. N. Clare

    2006-01-01

    Consumption of apples can provoke severe allergic reactions, in susceptible individuals, due to the presence of the allergen Mal d 3, a nonspecific lipid transfer protein, found largely in the fruit skin. Levels of Mal d 3 were determined in peel as a function of apple cultivar, position of the

  3. Antioxidative Effect of Seaweed Extracts in Chilled Storage of Minced Atlantic Mackerel (Scomber scombrus): Effect on Lipid and Protein Oxidation

    DEFF Research Database (Denmark)

    Babakhani, Aria; Farvin, K. H Sabeena; Jacobsen, Charlotte

    2016-01-01

    In this study, antioxidant activity of absolute ethanol, 50 % ethanol and water extracts of two species of seaweeds namely, Fucus serratus and Polysiphonia fucoides were evaluated for their ability to retard lipid and protein oxidation in minced mackerel. Mackerel mince added with 0.5 g/kg of ext...

  4. Collisions of droplets on spherical particles

    Science.gov (United States)

    Charalampous, Georgios; Hardalupas, Yannis

    2017-10-01

    Head-on collisions between droplets and spherical particles are examined for water droplets in the diameter range between 170 μm and 280 μm and spherical particles in the diameter range between 500 μm and 2000 μm. The droplet velocities range between 6 m/s and 11 m/s, while the spherical particles are fixed in space. The Weber and Ohnesorge numbers and ratio of droplet to particle diameter were between 92 deposition and splashing regimes, a regime is observed in the intermediate region, where the droplet forms a stable crown, which does not breakup but propagates along the particle surface and passes around the particle. This regime is prevalent when the droplets collide on small particles. The characteristics of the collision at the onset of rim instability are also described in terms of the location of the film on the particle surface and the orientation and length of the ejected crown. Proper orthogonal decomposition identified that the first 2 modes are enough to capture the overall morphology of the crown at the splashing threshold.

  5. Droplet generating device for droplet-based μTAS using electro-conjugate fluid

    Science.gov (United States)

    Iijima, Y.; Takemura, K.; Edamura, K.

    2017-05-01

    Droplet-based μTAS, which carries out biochemical inspection and synthesis by handling samples as droplets on a single chip, has been attracting attentions in recent years. Although miniaturization of a chip is progressed, there are some problems in miniaturization of a whole system because of the necessity to connect syringe pumps to the chip. Thus, this study aims to realize a novel droplets generating device for droplet-based μTAS using electro-conjugate fluid (ECF). The ECF is a dielectric liquid generating a powerful flow when subjected to high DC voltage. The ECF flow generation allows us to realize a tiny hydraulic power source. Using the ECF flow, we can develop a droplet generating device for droplet-based μTAS by placing minute electrode pairs in flow channels. The device contains two channels filled with the ECF, which are dispersed and continuous phases meeting at a T-junction. When a sample in the dispersed phase is injected by the ECF flow to the continuous phase at T-junction, droplets are generated by shearing force between the two phases. We conducted droplet generating experiment and confirmed that droplets are successfully generated when the flow rate of the continuous phase is between 90 and 360 mm3 s-1, and the flow rate of the dispersed phase is between 10 and 40 mm3 s-1. We also confirmed that the droplet diameter and the droplet production rate are controllable by tuning the applied voltage to the electrode pairs.

  6. Distribution of neutral lipids in the lipid droplet core

    DEFF Research Database (Denmark)

    Chaban, Vitaly V; Khandelia, Himanshu

    2014-01-01

    Cholesteryl esters (CEs) are a form of cholesterol (CHOL) storage in the living cells, as opposed to free CHOL. CEs are major constituents of low density lipoprotein particles. Therefore, CEs are implicated in provoking atherosclerosis. Arranged into cytoplasmic lipid droplets (LDs), CEs are stored...... intracellularly. They can also be transported extracellularly by means of lipoproteins. In this work, large-scale molecular dynamics (MD) simulations are used to characterize the molecular structure of LDs containing various fractions (10-50 mol %) of cholesteryl oleate (CO) with respect to triolein (TO) fraction...... the phospholipid interface, resulting from the structuring of hydrophilic groups. This structuring slowly decays in the direction toward the LD center of mass. No sorting of TO and CO is detected, irrespective of the molar fractions simulated. The distribution of CO within the LDs is significant in determining...

  7. Dynamics and molecular determinants of cytoplasmic lipid droplet clustering and dispersion.

    Directory of Open Access Journals (Sweden)

    David J Orlicky

    Full Text Available Perilipin-1 (Plin1, a prominent cytoplasmic lipid droplet (CLD binding phosphoprotein and key physiological regulator of triglyceride storage and lipolysis in adipocytes, is thought to regulate the fragmentation and dispersion of CLD that occurs in response to β-adrenergic activation of adenylate cyclase. Here we investigate the dynamics and molecular determinants of these processes using cell lines stably expressing recombinant forms of Plin1 and/or other members of the perilipin family. Plin1 and a C-terminal CLD-binding fragment of Plin1 (Plin1CT induced formation of single dense CLD clusters near the microtubule organizing center, whereas neither an N-terminal CLD-binding fragment of Plin1, nor Plin2 or Plin3 induced clustering. Clustered CLD coated by Plin1, or Plin1CT, dispersed in response to isoproterenol, or other agents that activate adenylate cyclase, in a process inhibited by the protein kinase A inhibitor, H89, and blocked by microtubule disruption. Isoproterenol-stimulated phosphorylation of CLD-associated Plin1 on serine 492 preceded their dispersion, and live cell imaging showed that cluster dispersion involved initial fragmentation of tight clusters into multiple smaller clusters, which then fragmented into well-dispersed individual CLD. siRNA knockdown of the cortical actin binding protein, moesin, induced disaggregation of tight clusters into multiple smaller clusters, and inhibited the reaggregation of dispersed CLD into tight clusters. Together these data suggest that the clustering and dispersion processes involve a complex orchestration of phosphorylation-dependent, microtubule-dependent and independent, and microfilament dependent steps.

  8. Dynamics and molecular determinants of cytoplasmic lipid droplet clustering and dispersion.

    Science.gov (United States)

    Orlicky, David J; Monks, Jenifer; Stefanski, Adrianne L; McManaman, James L

    2013-01-01

    Perilipin-1 (Plin1), a prominent cytoplasmic lipid droplet (CLD) binding phosphoprotein and key physiological regulator of triglyceride storage and lipolysis in adipocytes, is thought to regulate the fragmentation and dispersion of CLD that occurs in response to β-adrenergic activation of adenylate cyclase. Here we investigate the dynamics and molecular determinants of these processes using cell lines stably expressing recombinant forms of Plin1 and/or other members of the perilipin family. Plin1 and a C-terminal CLD-binding fragment of Plin1 (Plin1CT) induced formation of single dense CLD clusters near the microtubule organizing center, whereas neither an N-terminal CLD-binding fragment of Plin1, nor Plin2 or Plin3 induced clustering. Clustered CLD coated by Plin1, or Plin1CT, dispersed in response to isoproterenol, or other agents that activate adenylate cyclase, in a process inhibited by the protein kinase A inhibitor, H89, and blocked by microtubule disruption. Isoproterenol-stimulated phosphorylation of CLD-associated Plin1 on serine 492 preceded their dispersion, and live cell imaging showed that cluster dispersion involved initial fragmentation of tight clusters into multiple smaller clusters, which then fragmented into well-dispersed individual CLD. siRNA knockdown of the cortical actin binding protein, moesin, induced disaggregation of tight clusters into multiple smaller clusters, and inhibited the reaggregation of dispersed CLD into tight clusters. Together these data suggest that the clustering and dispersion processes involve a complex orchestration of phosphorylation-dependent, microtubule-dependent and independent, and microfilament dependent steps.

  9. Droplet trapping and fast acoustic release in a multi-height device with steady-state flow.

    Science.gov (United States)

    Rambach, Richard W; Linder, Kevin; Heymann, Michael; Franke, Thomas

    2017-10-11

    We demonstrate a novel multilayer polydimethylsiloxane (PDMS) device for selective storage and release of single emulsion droplets. Drops are captured in a microchannel cavity and can be released on-demand through a triggered surface acoustic wave pulse. The surface acoustic wave (SAW) is excited by a tapered interdigital transducer (TIDT) deposited on a piezoelectric lithium niobate (LiNbO 3 ) substrate and inverts the pressure difference across the cavity trap to push a drop out of the trap and back into the main flow channel. Droplet capture and release does not require a flow rate change, flow interruption, flow inversion or valve action and can be achieved in as fast as 20 ms. This allows both on-demand droplet capture for analysis and monitoring over arbitrary time scales, and continuous device operation with a high droplet rate of 620 drops per s. We hence decouple long-term droplet interrogation from other operations on the chip. This will ease integration with other microfluidic droplet operations and functional components.

  10. Equilibrium Droplets on Deformable Substrates: Equilibrium Conditions.

    Science.gov (United States)

    Koursari, Nektaria; Ahmed, Gulraiz; Starov, Victor M

    2018-05-15

    Equilibrium conditions of droplets on deformable substrates are investigated, and it is proven using Jacobi's sufficient condition that the obtained solutions really provide equilibrium profiles of both the droplet and the deformed support. At the equilibrium, the excess free energy of the system should have a minimum value, which means that both necessary and sufficient conditions of the minimum should be fulfilled. Only in this case, the obtained profiles provide the minimum of the excess free energy. The necessary condition of the equilibrium means that the first variation of the excess free energy should vanish, and the second variation should be positive. Unfortunately, the mentioned two conditions are not the proof that the obtained profiles correspond to the minimum of the excess free energy and they could not be. It is necessary to check whether the sufficient condition of the equilibrium (Jacobi's condition) is satisfied. To the best of our knowledge Jacobi's condition has never been verified for any already published equilibrium profiles of both the droplet and the deformable substrate. A simple model of the equilibrium droplet on the deformable substrate is considered, and it is shown that the deduced profiles of the equilibrium droplet and deformable substrate satisfy the Jacobi's condition, that is, really provide the minimum to the excess free energy of the system. To simplify calculations, a simplified linear disjoining/conjoining pressure isotherm is adopted for the calculations. It is shown that both necessary and sufficient conditions for equilibrium are satisfied. For the first time, validity of the Jacobi's condition is verified. The latter proves that the developed model really provides (i) the minimum of the excess free energy of the system droplet/deformable substrate and (ii) equilibrium profiles of both the droplet and the deformable substrate.

  11. Muscle Lipid Metabolism: Role of Lipid Droplets and Perilipins

    Directory of Open Access Journals (Sweden)

    Pablo Esteban Morales

    2017-01-01

    Full Text Available Skeletal muscle is one of the main regulators of carbohydrate and lipid metabolism in our organism, and therefore, it is highly susceptible to changes in glucose and fatty acid (FA availability. Skeletal muscle is an extremely complex tissue: its metabolic capacity depends on the type of fibers it is made up of and the level of stimulation it undergoes, such as acute or chronic contraction. Obesity is often associated with increased FA levels, which leads to the accumulation of toxic lipid intermediates, oxidative stress, and autophagy in skeletal fibers. This lipotoxicity is one of the most common causes of insulin resistance (IR. In this scenario, the “isolation” of certain lipids in specific cell compartments, through the action of the specific lipid droplet, perilipin (PLIN family of proteins, is conceived as a lifeguard compensatory strategy. In this review, we summarize the cellular mechanism underlying lipid mobilization and metabolism inside skeletal muscle, focusing on the function of lipid droplets, the PLIN family of proteins, and how these entities are modified in exercise, obesity, and IR conditions.

  12. Biochemical Aspects of a Serine Protease from Caesalpinia echinata Lam. (Brazilwood Seeds: A Potential Tool to Access the Mobilization of Seed Storage Proteins

    Directory of Open Access Journals (Sweden)

    Priscila Praxedes-Garcia

    2012-01-01

    Full Text Available Several proteins have been isolated from seeds of leguminous, but this is the first report that a protease was obtained from seeds of Caesalpinia echinata Lam., a tree belonging to the Fabaceae family. This enzyme was purified to homogeneity by hydrophobic interaction and anion exchange chromatographies and gel filtration. This 61-kDa serine protease (CeSP hydrolyses H-D-prolyl-L-phenylalanyl-L-arginine-p-nitroanilide (Km 55.7 μM in an optimum pH of 7.1, and this activity is effectively retained until 50∘C. CeSP remained stable in the presence of kosmotropic anions (PO4 3−, SO4 2−, and CH3COO− or chaotropic cations (K+ and Na+. It is strongly inhibited by TLCK, a serine protease inhibitor, but not by E-64, EDTA or pepstatin A. The characteristics of the purified enzyme allowed us to classify it as a serine protease. The role of CeSP in the seeds cannot be assigned yet but is possible to infer that it is involved in the mobilization of seed storage proteins.

  13. Tritium storage

    International Nuclear Information System (INIS)

    Hircq, B.

    1990-01-01

    This document represents a synthesis relative to tritium storage. After indicating the main storage particularities as regards tritium, storages under gaseous and solid form are after examined before establishing choices as a function of the main criteria. Finally, tritium storage is discussed regarding tritium devices associated to Fusion Reactors and regarding smaller devices [fr

  14. Serotonin storage pools in basophil leukemia and mast cells: characterization of two types of serotonin binding protein and radioautographic analysis of the intracellular distribution of [3H]serotonin

    International Nuclear Information System (INIS)

    Tamir, H.; Theoharides, T.C.; Gershon, M.D.; Askenase, P.W.

    1982-01-01

    The binding of serotonin to protein(s) derived from rat basophil leukemia (RBL) cells and mast cells was studied. Two types of serotonin binding protein in RBL cells was found. These proteins differed from one another in molecular weight and eluted in separate peaks from sephadex G-200 columns. Peak I protein (KD = 1.9 x 10 -6 M) was a glycoprotein that bound to concanavalin A (Con A); Peak II protein (KD 1 = 4.5 x 10 - 8 M; KD 2 = 3.9 x 10 -6 M) did not bind to Con A. Moreover, binding of [ 3 H]serotonin to protein of Peak I was sensitive to inhibition by reserpine, while binding of [ 3 H]serotonin to protein of Peak II resisted inhibition by that drug. Other differences between the two types of binding protein were found, the most significant of which was the far more vigorous conditions of homogenization required to extract Peak I than Peak II protein. Electron microscope radioautographic analysis of the intracellular distribution of [ 3 H] serotonin taken up in vitro by RBL cells or in vivo by murine mast cells indicated that essentially all of the labeled amine was located in cytoplasmic granules.No evidence for a pool in the cytosol was found and all granules were capable of becoming labeled. The presence of two types of intracellular serotonin binding proteins in these cells may indicate that there are two intracellular storage compartments for the amine. Both may be intragranular, but Peak I protein may be associated with the granular membrane while Peak II protein may be more free within the granular core. Different storage proteins may help to explain the differential release of amines from mast cell granules

  15. Evaluation of droplet deposition in rod bundle

    International Nuclear Information System (INIS)

    Ji, W.; Gu, C.Y.; Anglart, H.

    1997-01-01

    Deposition model for droplets in gas droplet two-phase flow in rod bundle is developed in this work using the Lagrangian method. The model is evaluated in a 9-rod bundle geometry. The deposition coefficient in the bundle geometry are compared with that in round tube. The influences of the droplet size and gas mass flow rate on deposition coefficient are investigated. Furthermore, the droplet motion is studied in more detail by dividing the bundle channel into sub-channels. The results show that the overall deposition coefficient in the bundle geometry is close to that in the round tube with the diameter equal to the bundle hydraulic diameter. The calculated deposition coefficient is found to be higher for higher gas mass flux and smaller droplets. The study in the sub-channels show that the ratio between the local deposition coefficient for a sub-channel and the averaged value for the whole bundle is close to a constant value, deviations from the mean value for all the calculated cases being within the range of ±13%. (author)

  16. Thermocapillary droplet actuation on structured solid surfaces

    Science.gov (United States)

    Karapetsas, George; Chamakos, Nikolaos T.; Papathanasiou, Athanasios G.

    2017-11-01

    The present work investigates, through 2D and 3D finite element simulations, the thermocapillary-driven flow inside a droplet which resides on a non-uniformly heated patterned surface. We employ a recently proposed sharp-interface scheme capable of efficiently modelling the flow over complicate surfaces and consider a wide range of substrate wettabilities, i.e. from hydrophilic to super-hydrophobic surfaces. Our simulations indicate that due to the presence of the solid structures and the induced effect of contact angle hysteresis, inherently predicted by our model, a critical thermal gradient arises beyond which droplet migration is possible, in line with previous experimental observations. The migration velocity as well as the direction of motion depends on the combined action of the net mechanical force along the contact line and the thermocapillary induced flow at the liquid-air interface. We also show that through a proper control and design of the substrate wettability, the contact angle hysteresis and the induced flow field it is possible to manipulate the droplet dynamics, e.g. controlling its motion along a predefined track or entrapping by a wetting defect a droplet based on its size as well as providing appropriate conditions for enhanced mixing inside the droplet. Funding from the European Research Council under the Europeans Community's Seventh Framework Programme (FP7/2007-2013)/ERC Grant agreement no. [240710] is acknowledged.

  17. Structural Transitions in Cholesteric Liquid Crystal Droplets

    Energy Technology Data Exchange (ETDEWEB)

    Zhou, Ye; Bukusoglu, Emre; Martínez-González, José A.; Rahimi, Mohammad; Roberts, Tyler F.; Zhang, Rui; Wang, Xiaoguang; Abbott, Nicholas L.; de Pablo, Juan J.

    2016-07-26

    Confinement of cholesteric liquid crystals (ChLC) into droplets leads to a delicate interplay between elasticity, chirality, and surface energy. In this work, we rely on a combination of theory and experiments to understand the rich morphological behavior that arises from that balance. More specifically, a systematic study of micrometer-sized ChLC droplets is presented as a function of chirality and surface energy (or anchoring). With increasing chirality, a continuous transition is observed from a twisted bipolar structure to a radial spherical structure, all within a narrow range of chirality. During such a transition, a bent structure is predicted by simulations and confirmed by experimental observations. Simulations are also able to capture the dynamics of the quenching process observed in experiments. Consistent with published work, it is found that nanoparticles are attracted to defect regions on the surface of the droplets. For weak anchoring conditions at the nanoparticle surface, ChLC droplets adopt a morphology similar to that of the equilibrium helical phase observed for ChLCs in the bulk. As the anchoring strength increases, a planar bipolar structure arises, followed by a morphological transition to a bent structure. The influence of chirality and surface interactions are discussed in the context of the potential use of ChLC droplets as stimuli-responsive materials for reporting molecular adsorbates.

  18. Statistical steady states in turbulent droplet condensation

    Science.gov (United States)

    Bec, Jeremie; Krstulovic, Giorgio; Siewert, Christoph

    2017-11-01

    We investigate the general problem of turbulent condensation. Using direct numerical simulations we show that the fluctuations of the supersaturation field offer different conditions for the growth of droplets which evolve in time due to turbulent transport and mixing. This leads to propose a Lagrangian stochastic model consisting of a set of integro-differential equations for the joint evolution of the squared radius and the supersaturation along droplet trajectories. The model has two parameters fixed by the total amount of water and the thermodynamic properties, as well as the Lagrangian integral timescale of the turbulent supersaturation. The model reproduces very well the droplet size distributions obtained from direct numerical simulations and their time evolution. A noticeable result is that, after a stage where the squared radius simply diffuses, the system converges exponentially fast to a statistical steady state independent of the initial conditions. The main mechanism involved in this convergence is a loss of memory induced by a significant number of droplets undergoing a complete evaporation before growing again. The statistical steady state is characterised by an exponential tail in the droplet mass distribution.

  19. Snap evaporation of droplets on smooth topographies.

    Science.gov (United States)

    Wells, Gary G; Ruiz-Gutiérrez, Élfego; Le Lirzin, Youen; Nourry, Anthony; Orme, Bethany V; Pradas, Marc; Ledesma-Aguilar, Rodrigo

    2018-04-11

    Droplet evaporation on solid surfaces is important in many applications including printing, micro-patterning and cooling. While seemingly simple, the configuration of evaporating droplets on solids is difficult to predict and control. This is because evaporation typically proceeds as a "stick-slip" sequence-a combination of pinning and de-pinning events dominated by static friction or "pinning", caused by microscopic surface roughness. Here we show how smooth, pinning-free, solid surfaces of non-planar topography promote a different process called snap evaporation. During snap evaporation a droplet follows a reproducible sequence of configurations, consisting of a quasi-static phase-change controlled by mass diffusion interrupted by out-of-equilibrium snaps. Snaps are triggered by bifurcations of the equilibrium droplet shape mediated by the underlying non-planar solid. Because the evolution of droplets during snap evaporation is controlled by a smooth topography, and not by surface roughness, our ideas can inspire programmable surfaces that manage liquids in heat- and mass-transfer applications.

  20. Lipid Droplet Formation Is Dispensable for Endoplasmic Reticulum-associated Degradation*

    OpenAIRE

    Olzmann, James A.; Kopito, Ron R.

    2011-01-01

    Proteins that fail to fold or assemble in the endoplasmic reticulum (ER) are destroyed by cytoplasmic proteasomes through a process known as ER-associated degradation. Substrates of this pathway are initially sequestered within the ER lumen and must therefore be dislocated across the ER membrane to be degraded. It has been proposed that generation of bicellar structures during lipid droplet formation may provide an “escape hatch” through which misfolded proteins, toxins, and viruses can exit ...

  1. Magnetic fluid droplet in a harmonic electric field

    Energy Technology Data Exchange (ETDEWEB)

    Kvasov, D., E-mail: kvasovdmitry@gmail.com [Lomonosov Moscow State University, Moscow (Russian Federation); Naletova, V. [Lomonosov Moscow State University, Moscow (Russian Federation); Beketova, E.; Dikanskii, Yu. [North-Caucasus Federal University, Stavropol (Russian Federation)

    2017-06-01

    A magnetic fluid droplet immersed in oil in an applied harmonic electric field is studied experimentally and theoretically. It is shown that deformations of the droplet observed experimentally are not described by the well-known theory. New double-layer droplet model which describes experimental data well is proposed. - Highlights: • The magnetic fluid droplet in the oil in a harmonic electric field is studied. • The paradoxical flattening effect of the droplet is observed experimentally. • For explaining this effect the model of the double-layer droplet is proposed. • Numerical and experimental data coincide qualitatively and quantitatively.

  2. Designed pneumatic valve actuators for controlled droplet breakup and generation.

    Science.gov (United States)

    Choi, Jae-Hoon; Lee, Seung-Kon; Lim, Jong-Min; Yang, Seung-Man; Yi, Gi-Ra

    2010-02-21

    The dynamic breakup of emulsion droplets was demonstrated in double-layered microfluidic devices equipped with designed pneumatic actuators. Uniform emulsion droplets, produced by shearing at a T-junction, were broken into smaller droplets when they passed downstream through constrictions formed by a pneumatically actuated valve in the upper control layer. The valve-assisted droplet breakup was significantly affected by the shape and layout of the control valves on the emulsion flow channel. Interestingly, by actuating the pneumatic valve immediately above the T-junction, the sizes of the emulsion droplets were controlled precisely in a programmatic manner that produced arrays of uniform emulsion droplets in various sizes and dynamic patterns.

  3. A cascade of iron-containing proteins governs the genetic iron starvation response to promote iron uptake and inhibit iron storage in fission yeast.

    Directory of Open Access Journals (Sweden)

    Javier Encinar del Dedo

    2015-03-01

    Full Text Available Iron is an essential cofactor, but it is also toxic at high levels. In Schizosaccharomyces pombe, the sensor glutaredoxin Grx4 guides the activity of the repressors Php4 and Fep1 to mediate a complex transcriptional response to iron deprivation: activation of Php4 and inactivation of Fep1 leads to inhibition of iron usage/storage, and to promotion of iron import, respectively. However, the molecular events ruling the activity of this double-branched pathway remained elusive. We show here that Grx4 incorporates a glutathione-containing iron-sulfur cluster, alone or forming a heterodimer with the BolA-like protein Fra2. Our genetic study demonstrates that Grx4-Fra2, but not Fep1 nor Php4, participates not only in iron starvation signaling but also in iron-related aerobic metabolism. Iron-containing Grx4 binds and inactivates the Php4 repressor; upon iron deprivation, the cluster in Grx4 is probably disassembled, the proteins dissociate, and Php4 accumulates at the nucleus and represses iron consumption genes. Fep1 is also an iron-containing protein, and the tightly bound iron is required for transcriptional repression. Our data suggest that the cluster-containing Grx4-Fra2 heterodimer constitutively binds to Fep1, and upon iron deprivation the disassembly of the iron cluster between Grx4 and Fra2 promotes reverse metal transfer from Fep1 to Grx4-Fra2, and de-repression of iron-import genes. Our genetic and biochemical study demonstrates that the glutaredoxin Grx4 independently governs the Php4 and Fep1 repressors through metal transfer. Whereas iron loss from Grx4 seems to be sufficient to release Php4 and allow its nuclear accumulation, total or partial disassembly of the Grx4-Fra2 cluster actively participates in iron-containing Fep1 activation by sequestering its iron and decreasing its interaction with promoters.

  4. Use of egg white protein powder based films fortified with sage and lemon balm essential oils in the storage of lor cheese

    Directory of Open Access Journals (Sweden)

    Gökhan Kavas

    2016-03-01

    Full Text Available Edible film was produced by adding 3 % sorbitol (w/v to egg white protein powder (EWPP. The first group of lor cheese samples was coated with a film fortified by sage essential oil (SEO and the second group of samples was coated with films enriched by adding lemon balm essential oil (BEO at various concentrations [0.5 %, 1 %, 2 % (v/v]. The films were labeled as EWPPSEO(0.5, EWPPSEO(1, EWPPSEO(2, EWPPBEO(0.5, EWPPBEO(1, EWPPBEO(2 to indicate the type and the concentration of the additive. The third batch of the lor cheese samples was coated exclusively with non-fortified EWPP and the fourth batch was uncoated. All of the cheese samples were artificially contaminated with Escherichia coli O157:H7 (E. coli O157:H7, Listeria monocytogenes (L. monocytogenes and Staphylococcus aureus (S. aureus. Viable cell counts of these species, yeasts and moulds were determined after the cheese production. All the samples were stored at +4 °C. Their physicochemical and microbiological properties were examined on the 1st, 7th, 15th and 30th day of the storage. Thereat significant (P0.05. Physicochemical and antibacterial properties were more significant in SEO at all concentrations compared to BEO. However, the antifungal effect of BEO was higher than that of SEO. The antifungal effect of BEO was the same at 1 % (v/v and 2 % (v/v concentrations. E. coli O157:H7 was the most resistant microorganism to the essential oils while L. monocytogenes was the most sensitive. EWPP showed a bacteriostatic effect on the microorganisms and bactericidal effects were determined on the 30th day of the storage against L. monocytogenes and yeast-moulds.

  5. Predicting protein aggregation during storage in lyophilized solids using solid state amide hydrogen/deuterium exchange with mass spectrometric analysis (ssHDX-MS).

    Science.gov (United States)

    Moorthy, Balakrishnan S; Schultz, Steven G; Kim, Sherry G; Topp, Elizabeth M

    2014-06-02

    Solid state amide hydrogen/deuterium exchange with mass spectrometric analysis (ssHDX-MS) was used to assess the conformation of myoglobin (Mb) in lyophilized formulations, and the results correlated with the extent of aggregation during storage. Mb was colyophilized with sucrose (1:1 or 1:8 w/w), mannitol (1:1 w/w), or NaCl (1:1 w/w) or in the absence of excipients. Immediately after lyophilization, samples of each formulation were analyzed by ssHDX-MS and Fourier transform infrared spectroscopy (FTIR) to assess Mb conformation, and by dynamic light scattering (DLS) and size exclusion chromatography (SEC) to determine the extent of aggregation. The remaining samples were then placed on stability at 25 °C and 60% RH or 40 °C and 75% RH for up to 1 year, withdrawn at intervals, and analyzed for aggregate content by SEC and DLS. In ssHDX-MS of samples immediately after lyophilization (t = 0), Mb was less deuterated in solids containing sucrose (1:1 and 1:8 w/w) than in those containing mannitol (1:1 w/w), NaCl (1:1 w/w), or Mb alone. Deuterium uptake kinetics and peptide mass envelopes also indicated greater Mb structural perturbation in mannitol, NaCl, or Mb-alone samples at t = 0. The extent of deuterium incorporation and kinetic parameters related to rapidly and slowly exchanging amide pools (Nfast, Nslow), measured at t = 0, were highly correlated with the extent of aggregation on storage as measured by SEC. In contrast, the extent of aggregation was weakly correlated with FTIR band intensity and peak position measured at t = 0. The results support the use of ssHDX-MS as a formulation screening tool in developing lyophilized protein drug products.

  6. Droplet condensation in rapidly decaying pressure fields

    International Nuclear Information System (INIS)

    Peterson, P.F.; Bai, R.Y.; Schrock, V.E.; Hijikata, K.

    1992-01-01

    Certain promising schemes for cooling inertial confinement fusion reactors call for highly transient condensation in a rapidly decaying pressure field. After an initial period of condensation on a subcooled droplet, undesirable evaporation begins to occur. Recirculation within the droplet strongly impacts the character of this condensation-evaporation cycle, particularly when the recirculation time constant is of the order of the pressure decay time constant. Recirculation can augment the heat transfer, delay the onset of evaporation, and increase the maximum superheat inside the drop by as much as an order of magnitude. This numerical investigation identifies the most important parameters and physics characterizing transient, high heat flux droplet condensation. The results can be applied to conceptual designs of inertial confinement fusion reactors, where initial temperature differences on the order of 1,500 K decay to zero over time spans the order of tens of milliseconds

  7. Lattice-Boltzmann simulations of droplet evaporation

    KAUST Repository

    Ledesma-Aguilar, Rodrigo; Vella, Dominic; Yeomans, Julia M.

    2014-01-01

    © the Partner Organisations 2014. We study the utility and validity of lattice-Boltzmann (LB) simulations to explore droplet evaporation driven by a concentration gradient. Using a binary-fluid lattice-Boltzmann algorithm based on Cahn-Hilliard dynamics, we study the evaporation of planar films and 3D sessile droplets from smooth solid surfaces. Our results show that LB simulations accurately reproduce the classical regime of quasi-static dynamics. Beyond this limit, we show that the algorithm can be used to explore regimes where the evaporative and diffusive timescales are not widely separated, and to include the effect of boundaries of prescribed driving concentration. We illustrate the method by considering the evaporation of a droplet from a solid surface that is chemically patterned with hydrophilic and hydrophobic stripes. This journal is

  8. Lattice-Boltzmann simulations of droplet evaporation

    KAUST Repository

    Ledesma-Aguilar, Rodrigo

    2014-09-04

    © the Partner Organisations 2014. We study the utility and validity of lattice-Boltzmann (LB) simulations to explore droplet evaporation driven by a concentration gradient. Using a binary-fluid lattice-Boltzmann algorithm based on Cahn-Hilliard dynamics, we study the evaporation of planar films and 3D sessile droplets from smooth solid surfaces. Our results show that LB simulations accurately reproduce the classical regime of quasi-static dynamics. Beyond this limit, we show that the algorithm can be used to explore regimes where the evaporative and diffusive timescales are not widely separated, and to include the effect of boundaries of prescribed driving concentration. We illustrate the method by considering the evaporation of a droplet from a solid surface that is chemically patterned with hydrophilic and hydrophobic stripes. This journal is

  9. Droplet sizes, dynamics and deposition in vertical annular flow

    International Nuclear Information System (INIS)

    Lopes, J.C.B.; Dukler, A.E.

    1985-10-01

    The role of droplets in vertical upwards annular flow is investigated, focusing on the droplet size distributions, dynamics, and deposition phenomena. An experimental program was performed based on a new laser optical technique developed in these laboratories and implemented here for annular flow. This permitted the simultaneous measurement of droplet size, axial and radial velocity. The dependence of droplet size distributions on flow conditions is analyzed. The Upper-Log Normal function proves to be a good model for the size distribution. The mechanism controlling the maximum stable drop size was found to result from the interaction of the pressure fluctuations of the turbulent flow of the gas core with the droplet. The average axial droplet velocity showed a weak dependence on gas rates. This can be explained once the droplet size distribution and droplet size-velocity relationship are analyzed simultaneously. The surprising result from the droplet conditional analysis is that larger droplet travel faster than smaller ones. This dependence cannot be explained if the drag curves used do not take into account the high levels of turbulence present in the gas core in annular flow. If these are considered, then interesting new situations of multiplicity and stability of droplet terminal velocities are encountered. Also, the observed size-velocity relationship can be explained. A droplet deposition is formulated based on the particle inertia control. This permitted the calculation of rates of drop deposition directly from the droplet size and velocities data

  10. Interface-Resolving Simulation of Collision Efficiency of Cloud Droplets

    Science.gov (United States)

    Wang, Lian-Ping; Peng, Cheng; Rosa, Bodgan; Onishi, Ryo

    2017-11-01

    Small-scale air turbulence could enhance the geometric collision rate of cloud droplets while large-scale air turbulence could augment the diffusional growth of cloud droplets. Air turbulence could also enhance the collision efficiency of cloud droplets. Accurate simulation of collision efficiency, however, requires capture of the multi-scale droplet-turbulence and droplet-droplet interactions, which has only been partially achieved in the recent past using the hybrid direct numerical simulation (HDNS) approach. % where Stokes disturbance flow is assumed. The HDNS approach has two major drawbacks: (1) the short-range droplet-droplet interaction is not treated rigorously; (2) the finite-Reynolds number correction to the collision efficiency is not included. In this talk, using two independent numerical methods, we will develop an interface-resolved simulation approach in which the disturbance flows are directly resolved numerically, combined with a rigorous lubrication correction model for near-field droplet-droplet interaction. This multi-scale approach is first used to study the effect of finite flow Reynolds numbers on the droplet collision efficiency in still air. Our simulation results show a significant finite-Re effect on collision efficiency when the droplets are of similar sizes. Preliminary results on integrating this approach in a turbulent flow laden with droplets will also be presented. This work is partially supported by the National Science Foundation.

  11. Fluorescence detection system for microfluidic droplets

    Science.gov (United States)

    Chen, Binyu; Han, Xiaoming; Su, Zhen; Liu, Quanjun

    2018-05-01

    In microfluidic detection technology, because of the universality of optical methods in laboratory, optical detection is an attractive solution for microfluidic chip laboratory equipment. In addition, the equipment with high stability and low cost can be realized by integrating appropriate optical detection technology on the chip. This paper reports a detection system for microfluidic droplets. Photomultiplier tubes (PMT) is used as a detection device to improve the sensitivity of detection. This system improves the signal to noise ratio by software filtering and spatial filter. The fluorescence intensity is proportional to the concentration of the fluorescence and intensity of the laser. The fluorescence micro droplets of different concentrations can be distinguished by this system.

  12. Droplets bouncing on a standing wave field

    Science.gov (United States)

    Pucci, Giuseppe; Tambasco, Lucas; Harris, Daniel; Bush, John

    2017-11-01

    A liquid bath subject to a vertical vibration becomes unstable to standing surface waves at a critical vibrational acceleration known as the Faraday threshold. We examine the behavior of a millimetric droplet bouncing on the surface of a quasi-one-dimensional fluid channel above the Faraday threshold. We identify a sequence of bifurcations that occurs as the vibrational acceleration is increased progressively, ultimately leading to the erratic, diffusive motion of the droplet along the length of the channel. A simple theoretical model is presented. This work was supported by the US National Science Foundation through Grants CMMI-1333242 and DMS-1614043.

  13. CRP-Mediated Carbon Catabolite Regulation of Yersinia pestis Biofilm Formation Is Enhanced by the Carbon Storage Regulator Protein, CsrA.

    Directory of Open Access Journals (Sweden)

    Stephan P Willias

    Full Text Available The natural transmission of Yersinia pestis is reliant upon biofilm blockage of the flea vector. However, the environmentally-responsive adaptive regulators which facilitate Y. pestis biofilm production in accordance with the flea midgut milieu are not well understood. We seek to establish the impact of available carbon source metabolism and storage upon Y. pestis biofilm production. Our findings demonstrate that Y. pestis biofilm production is subject to carbon catabolite regulation in which the presence of glucose impairs biofilm production; whereas, the sole metabolism of alternate carbon sources promotes robust biofilm formation. This observation is facilitated by the cAMP receptor protein, CRP. In accordance with a stark growth defect, deletion of crp in both CO92 and KIM6+ Y. pestis strains significantly impaired biofilm production when solely utilizing alternate carbon sources. Media supplementation with cAMP, a small-molecule activator of CRP, did not significantly alter Y. pestis biofilm production. Furthermore, CRP did not alter mRNA abundance of previously-characterized hms biofilm synthesis and regulation factors. Therefore, our findings indicate CRP does not confer a direct stimulatory effect, but may indirectly promote Y. pestis biofilm production by facilitating the alternate carbon source expression profile. Additionally, we assessed the impact of the carbon storage regulator protein, CsrA, upon Y. pestis biofilm production. Contrary to what has been described for E. coli, Y. pestis biofilm formation was found to be enhanced by CsrA. Regardless of media composition and available carbon source, deletion of csrA significantly impaired Y. pestis biofilm production. CsrA was found to promote Y. pestis biofilm production independent of glycogen regulation. Loss of csrA did not significantly alter relative hmsH, hmsP, or hmsT mRNA abundance. However, deletion of hmsP in the csrA-deficient mutant enabled excessive biofilm production

  14. TRAJECTORY AND INCINERATION OF ROGUE DROPLETS IN A TURBULENT DIFFUSION FLAME

    Science.gov (United States)

    The trajectory and incineration efficiency of individual droplet streams of a fuel mixture injected into a swirling gas turbulent diffusion flame were measured as a function of droplet size, droplet velocity, interdroplet spacing, and droplet injection angle. Additional experimen...

  15. Evaluating the capabilities and uncertainties of droplet measurements for the fog droplet spectrometer (FM-100

    Directory of Open Access Journals (Sweden)

    J. K. Spiegel

    2012-09-01

    Full Text Available Droplet size spectra measurements are crucial to obtain a quantitative microphysical description of clouds and fog. However, cloud droplet size measurements are subject to various uncertainties. This work focuses on the error analysis of two key measurement uncertainties arising during cloud droplet size measurements with a conventional droplet size spectrometer (FM-100: first, we addressed the precision with which droplets can be sized with the FM-100 on the basis of the Mie theory. We deduced error assumptions and proposed a new method on how to correct measured size distributions for these errors by redistributing the measured droplet size distribution using a stochastic approach. Second, based on a literature study, we summarized corrections for particle losses during sampling with the FM-100. We applied both corrections to cloud droplet size spectra measured at the high alpine site Jungfraujoch for a temperature range from 0 °C to 11 °C. We showed that Mie scattering led to spikes in the droplet size distributions using the default sizing procedure, while the new stochastic approach reproduced the ambient size distribution adequately. A detailed analysis of the FM-100 sampling efficiency revealed that particle losses were typically below 10% for droplet diameters up to 10 μm. For larger droplets, particle losses can increase up to 90% for the largest droplets of 50 μm at ambient wind speeds below 4.4 m s−1 and even to >90% for larger angles between the instrument orientation and the wind vector (sampling angle at higher wind speeds. Comparisons of the FM-100 to other reference instruments revealed that the total liquid water content (LWC measured by the FM-100 was more sensitive to particle losses than to re-sizing based on Mie scattering, while the total number concentration was only marginally influenced by particle losses. Consequently, for further LWC measurements with the FM-100 we strongly recommend to consider (1 the

  16. Control of charged droplets using electrohydrodynamic repulsion for circular droplet patterning

    International Nuclear Information System (INIS)

    Kim, Bumjoo; Sung, Jungwoo; Lim, Geunbae; Nam, Hyoryung; Kim, Sung Jae; Joo, Sang W

    2011-01-01

    We report a novel method to form a circular pattern of monodisperse microdroplets using an electrohydrodynamic repulsion (EDR) mechanism. EDR is a phenomenon of electrostatical bounced microdroplets from an accumulated droplet on a bottom substrate. In addition to a regular EDR system, by placing a ring electrode between the capillary and ground substrate, two separate regions were created. A parameter study of two regions was carried out for droplet formation and falling velocity to control the radius of the generated droplets and the circular patterns independently. Based on energy conservation theory, our experimental results showed that the free-falling region exerted crucial influences on the sizes of the circular patterns

  17. Velocity and rotation measurements in acoustically levitated droplets

    Energy Technology Data Exchange (ETDEWEB)

    Saha, Abhishek [University of Central Florida, Orlando, FL 32816 (United States); Basu, Saptarshi [Indian Institute of Science, Bangalore 560012 (India); Kumar, Ranganathan, E-mail: ranganathan.kumar@ucf.edu [University of Central Florida, Orlando, FL 32816 (United States)

    2012-10-01

    The velocity scale inside an acoustically levitated droplet depends on the levitator and liquid properties. Using Particle Imaging Velocimetry (PIV), detailed velocity measurements have been made in a levitated droplet of different diameters and viscosity. The maximum velocity and rotation are normalized using frequency and amplitude of acoustic levitator, and droplet viscosity. The non-dimensional data are fitted for micrometer- and millimeter-sized droplets levitated in different levitators for different viscosity fluids. It is also shown that the rotational speed of nanosilica droplets at an advanced stage of vaporization compares well with that predicted by exponentially fitted parameters. -- Highlights: ► Demonstrates the importance of rotation in a levitated droplet that leads to controlled morphology. ► Provides detailed measurements of Particle Image Velocimetry inside levitated droplets. ► Shows variation of vortex strength with the droplet diameter and viscosity of the liquid.

  18. Velocity and rotation measurements in acoustically levitated droplets

    International Nuclear Information System (INIS)

    Saha, Abhishek; Basu, Saptarshi; Kumar, Ranganathan

    2012-01-01

    The velocity scale inside an acoustically levitated droplet depends on the levitator and liquid properties. Using Particle Imaging Velocimetry (PIV), detailed velocity measurements have been made in a levitated droplet of different diameters and viscosity. The maximum velocity and rotation are normalized using frequency and amplitude of acoustic levitator, and droplet viscosity. The non-dimensional data are fitted for micrometer- and millimeter-sized droplets levitated in different levitators for different viscosity fluids. It is also shown that the rotational speed of nanosilica droplets at an advanced stage of vaporization compares well with that predicted by exponentially fitted parameters. -- Highlights: ► Demonstrates the importance of rotation in a levitated droplet that leads to controlled morphology. ► Provides detailed measurements of Particle Image Velocimetry inside levitated droplets. ► Shows variation of vortex strength with the droplet diameter and viscosity of the liquid.

  19. Charge Transfer into Aqueous Droplets via Kilovolt Potentials

    Science.gov (United States)

    Hamlin, B. S.; Rosenberg, E. R.; Ristenpart, W. D.

    2012-11-01

    When an aqueous droplet immersed in an insulating oil contacts an electrified surface, the droplet acquires net charge. For sufficiently large field strengths, the charged droplet is driven back and forth electrophoretically between the electrodes, in essence ``bouncing'' between them. Although it is clear that the droplet acquires charge, the underlying mechanism controlling the charge transfer process has been unclear. Here we demonstrate that the chemical species present in the droplet strongly affect the charge transfer process into the drop. Using two independent charge measurement techniques, high speed video velocimetry and direct current measurement, we show that the charge acquired during contact is strongly influenced by the droplet pH. We also provide physical evidence that the electrodes undergo electroplating or corrosion for droplets with appropriate chemical species present. Together, the observations strongly suggest that electrochemical reactions govern the charge transfer process into the droplet.

  20. Encapsulation of emulsion droplets by organo–silica shells

    NARCIS (Netherlands)

    Zoldesi, C.; Steegstra, Patrick; Imhof, Arnout

    2007-01-01

    Surfactant-stabilized emulsion droplets were used as templates for the synthesis of hollow colloidal particles. Monodisperse silicone oil droplets were prepared by hydrolysis and polymerization of dimethyldiethoxysiloxane monomer, in the presence of surfactant: sodium dodecyl sulphate (SDS, anionic)

  1. Microfluidic droplet generator with controlled break-up mechanism

    KAUST Repository

    Gonzalez, David Conchouso

    2017-04-13

    Droplet generation devices and systems that parallelize droplet generation devices are provided. The droplet generation devices can include a symmetric block-and-break system and a tapered droplet generation zone. The symmetric block-and-break system can include a pair of break channels and a pair of bypass channels symmetrically arranged with respect to the dispersed-phase input channel and the output channel. The droplet generation devices can generate monodisperse droplets with a predefined volume over a range of flow rates, pressures, and fluid properties. The droplet generation devices are therefore capable of parallelization to achieve large-capacity droplet generation, e.g. greater than 1 L/hr, with small overall coefficients of variation.

  2. Thermophoretically driven water droplets on graphene and boron nitride surfaces

    Science.gov (United States)

    Rajegowda, Rakesh; Kannam, Sridhar Kumar; Hartkamp, Remco; Sathian, Sarith P.

    2018-05-01

    We investigate thermally driven water droplet transport on graphene and hexagonal boron nitride (h-BN) surfaces using molecular dynamics simulations. The two surfaces considered here have different wettabilities with a significant difference in the mode of droplet transport. The water droplet travels along a straighter path on the h-BN sheet than on graphene. The h-BN surface produced a higher driving force on the droplet than the graphene surface. The water droplet is found to move faster on h-BN surface compared to graphene surface. The instantaneous contact angle was monitored as a measure of droplet deformation during thermal transport. The characteristics of the droplet motion on both surfaces is determined through the moment scaling spectrum. The water droplet on h-BN surface showed the attributes of the super-diffusive process, whereas it was sub-diffusive on the graphene surface.

  3. A novel coarsening mechanism of droplets in immiscible fluid mixtures

    Science.gov (United States)

    Shimizu, Ryotaro; Tanaka, Hajime

    2015-06-01

    In our daily lives, after shaking a salad dressing, we see the coarsening of oil droplets suspended in vinegar. Such a demixing process is observed everywhere in nature and also of technological importance. For a case of high droplet density, domain coarsening proceeds with inter-droplet collisions and the resulting coalescence. This phenomenon has been explained primarily by the so-called Brownian-coagulation mechanism: stochastic thermal forces exerted by molecules induce random motion of individual droplets, causing accidental collisions and subsequent interface-tension-driven coalescence. Contrary to this, here we demonstrate that the droplet motion is not random, but hydrodynamically driven by the composition Marangoni force due to an interfacial tension gradient produced in each droplet as a consequence of composition correlation among droplets. This alters our physical understanding of droplet coarsening in immiscible liquid mixtures on a fundamental level.

  4. The Evaporation of Liquid Droplets in Highly Turbulent Gas Streams

    National Research Council Canada - National Science Library

    Gould, Richard

    1998-01-01

    Single acetone and heptane droplets were suspended from a hypodermic needle in turbulent airflow, and the Nusselt number was obtained from direct measurements of the droplet diameter and evaporation rate...

  5. Flow field induced particle accumulation inside droplets in rectangular channels.

    Science.gov (United States)

    Hein, Michael; Moskopp, Michael; Seemann, Ralf

    2015-07-07

    Particle concentration is a basic operation needed to perform washing steps or to improve subsequent analysis in many (bio)-chemical assays. In this article we present field free, hydrodynamic accumulation of particles and cells in droplets flowing within rectangular micro-channels. Depending on droplet velocity, particles either accumulate at the rear of the droplet or are dispersed over the entire droplet cross-section. We show that the observed particle accumulation behavior can be understood by a coupling of particle sedimentation to the internal flow field of the droplet. The changing accumulation patterns are explained by a qualitative change of the internal flow field. The topological change of the internal flow field, however, is explained by the evolution of the droplet shape with increasing droplet velocity altering the friction with the channel walls. In addition, we demonstrate that accumulated particles can be concentrated, removing excess dispersed phase by splitting the droplet at a simple channel junction.

  6. Sensitive and predictable separation of microfluidic droplets by size using in-line passive filter.

    Science.gov (United States)

    Ding, Ruihua; Ung, W Lloyd; Heyman, John A; Weitz, David A

    2017-01-01

    Active manipulation of droplets is crucial in droplet microfluidics. However, droplet polydispersity decreases the accuracy of active manipulation. We develop a microfluidic "droplet filter" that accurately separates droplets by size. The droplet filter has a sharp size cutoff and is capable of distinguishing droplets differing in volume by 20%. A simple model explains the behavior of the droplets as they pass through the filter. We show application of the filter in improving dielectric sorting efficiency.

  7. Droplet Traffic at a Simple Junction at Low Capillary Numbers

    Science.gov (United States)

    Engl, Wilfried; Roche, Matthieu; Colin, Annie; Panizza, Pascal; Ajdari, Armand

    2005-11-01

    We report that, when a train of confined droplets flowing through a channel reaches a junction, the droplets either are alternately distributed between the different outlets or all collect into the shortest one. We argue that this behavior is due to the hydrodynamic feedback of droplets in the different outlets on the selection process occurring at the junction. A “mean field” model, yielding semiquantitative results, offers a first guide to predict droplet traffic in branched networks.

  8. Micrometer-sized TPM emulsion droplets with surface-mobile binding groups

    Science.gov (United States)

    van der Wel, Casper; van de Stolpe, Guido L.; Verweij, Ruben W.; Kraft, Daniela J.

    2018-03-01

    Colloids coated with lipid membranes have been widely employed for fundamental studies of lipid membrane processes, biotechnological applications such as drug delivery and biosensing, and more recently, for self-assembly. The latter has been made possible by inserting DNA oligomers with covalently linked hydrophobic anchors into the membrane. The lateral mobility of the DNA linkers on micrometer-sized droplets and solid particles has opened the door to creating structures with unprecedented structural flexibility. Here, we investigate micro-emulsions of TPM (3-(trimethoxysilyl)propyl methacrylate) as a platform for lipid monolayers and further functionalization with proteins and DNA oligonucleotides. TPM droplets can be produced with a narrow size distribution and are polymerizable, thus providing supports for model lipid membranes with controlled size and curvature. With fluorescence recovery after photobleaching, we observed that droplet-attached lipids, NeutrAvidin proteins, as well as DNA oligonucleotides all show mobility on the surface. We explored the assembly of micron-sized particles on TPM-droplets by exploiting either avidin-biotin interactions or double-stranded DNA with complementary single-stranded end groups. While the single molecules are mobile, the particles that are attached to them are not. We propose that this is caused by the heterogeneous nature of emulsified TPM, which forms an oligomer network that limits the collective motion of linkers, but allows the surface mobility of individual molecules.

  9. Energy storage

    Science.gov (United States)

    Kaier, U.

    1981-04-01

    Developments in the area of energy storage are characterized, with respect to theory and laboratory, by an emergence of novel concepts and technologies for storing electric energy and heat. However, there are no new commercial devices on the market. New storage batteries as basis for a wider introduction of electric cars, and latent heat storage devices, as an aid for solar technology applications, with satisfactory performance standards are not yet commercially available. Devices for the intermediate storage of electric energy for solar electric-energy systems, and for satisfying peak-load current demands in the case of public utility companies are considered. In spite of many promising novel developments, there is yet no practical alternative to the lead-acid storage battery. Attention is given to central heat storage for systems transporting heat energy, small-scale heat storage installations, and large-scale technical energy-storage systems.

  10. Influence of calcium, magnesium, or potassium ions on the formation and stability of emulsions prepared using highly hydrolyzed whey proteins.

    Science.gov (United States)

    Ramkumar, C; Singh, H; Munro, P A; Singh, A M

    2000-05-01

    Oil-in-water emulsions (4 wt % soy oil) containing 4 wt % whey protein hydrolysate (WPH) (27% degree of hydrolysis) and different levels of calcium, magnesium, or potassium chloride were prepared in a two-stage homogenizer. Other emulsions containing 4 wt % WPH but including 0.35 wt % hydroxylated lecithin and different levels of the above minerals were similarly prepared. The formation and stability of these emulsions were determined by measuring oil droplet size distributions using laser light scattering and by confocal scanning laser microscopy and a gravity creaming test. Both lecithin-free and lecithin-containing emulsions showed no change in droplet size distributions with increasing concentration of potassium in the range 0-37.5 mM. In contrast, the diameter of emulsion droplets increased with increasing calcium or magnesium concentration >12.5 mM. Emulsions containing hydroxylated lecithin were more sensitive to the addition of calcium or magnesium than the lecithin-free emulsions. Storage of emulsions at 20 degrees C for 24 h further increased the diameter of droplets and resulted in extensive creaming in emulsions containing >25 mM calcium or magnesium. It appears that both flocculation and coalescence processes were involved in the destabilization of emulsions induced by the addition of divalent cations.

  11. Influence of film dimensions on film droplet formation.

    Science.gov (United States)

    Holmgren, Helene; Ljungström, Evert

    2012-02-01

    Aerosol particles may be generated from rupturing liquid films through a droplet formation mechanism. The present work was undertaken with the aim to throw some light on the influence of film dimensions on droplet formation with possible consequences for exhaled breath aerosol formation. The film droplet formation process was mimicked by using a purpose-built device, where fluid films were spanned across holes of known diameters. As the films burst, droplets were formed and the number and size distributions of the resulting droplets were determined. No general relation could be found between hole diameter and the number of droplets generated per unit surface area of fluid film. Averaged over all film sizes, a higher surface tension yielded higher concentrations of droplets. Surface tension did not influence the resulting droplet diameter, but it was found that smaller films generated smaller droplets. This study shows that small fluid films generate droplets as efficiently as large films, and that droplets may well be generated from films with diameters below 1 mm. This has implications for the formation of film droplets from reopening of closed airways because human terminal bronchioles are of similar dimensions. Thus, the results provide support for the earlier proposed mechanism where reopening of closed airways is one origin of exhaled particles.

  12. Dynamics of droplet breakup in a T-junction

    NARCIS (Netherlands)

    Hoang, D.A.; Portela, L.M.; Kleijn, C.R.; Kreutzer, M.T.; Van Steijn, V.

    2013-01-01

    The breakup of droplets due to creeping motion in a confined microchannel geometry is studied using three-dimensional numerical simulations. Analogously to unconfined droplets, there exist two distinct breakup phases: (i) a quasi-steady droplet deformation driven by the externally applied flow; and

  13. Both dioscorin, the tuber storage protein of yam (Dioscorea alata cv. Tainong No. 1), and its peptic hydrolysates exhibited angiotensin converting enzyme inhibitory activities.

    Science.gov (United States)

    Hsu, Feng-Lin; Lin, Yaw-Huei; Lee, Mei-Hsien; Lin, Chien-Liang; Hou, Wen-Chi

    2002-10-09

    Dioscorin, the tuber storage protein of yam (Dioscorea alata cv. Tainong No. 1), was purified to homogeneity by DE-52 ion-exchange chromatography. This purified dioscorin was shown by spectrophotometric methods to inhibit angiotensin converting enzyme (ACE) in a dose-dependent manner (12.5-750 microg, respectively, 20.83-62.5% inhibitions) using N-[3-(2-furyl)acryloyl]-Phe-Gly-Gly (FAPGG) as substrates. The 50% inhibition (IC(50)) of ACE activity was 6.404 microM dioscorin (250 microg corresponding to 7.81 nmol) compared to that of 0.00781 microM (0.0095 nmol) for captopril. The commercial bovine serum albumin and casein (bovine milk) showed less ACE inhibitory activity. The use of qualitative TLC also showed dioscorin as ACE inhibitors. Dioscorin showed mixed noncompetitive inhibitions against ACE; when 31.25 microg of dioscorin (0.8 microM) was added, the apparent inhibition constant (K(i)) was 2.738 microM. Pepsin was used for dioscorin hydrolysis at 37 degrees C for different times. It was found that the ACE inhibitory activity was increased from 51.32% to about 75% during 32 h hydrolysis. The smaller peptides were increased with increasing pepsin hydrolytic times. Dioscorin and its hydrolysates might be a potential for hypertension control when people consume yam tuber.

  14. Effects of oral administration of yam tuber storage protein, dioscorin, to BALB/c mice for 21-days on immune responses.

    Science.gov (United States)

    Liu, Yen-Wenn; Liu, Ju-Chi; Huang, Chun-Yao; Wang, Chung-Kwe; Shang, Huey-Fang; Hou, Wen-Chi

    2009-10-14

    Dioscorin, the tuber storage protein of yam, was reported to have immunomodulatory activity in RAW264.7 murine macrophage cell lines ( Food and Chemical Toxicology , 2007 , 45 , 2312 -2318 ). However, the immunomodulatory function of dioscorin after being ingested was not elucidated in vivo. Hence, BALB/c mice were given oral dioscorin (2.5 and 20 mg/kg/day) once a day for 21-days. Lymphocyte subpopulation changes in the peripheral blood and splenocytes, stimulation in phagocytosis of the polymorphonuclear cell (PMN) and monocytes, the natural killer (NK) cell cytotoxicity, the splenocyte proliferation, and cytokine secretions in the presence of PHA were determined. The number changes of Peyer's patches and secreted IgA (sIgA) in the feces were determined. Oral dioscorin for 21-days increased the subpopulation in natural killer cells (CD49(+)) and/or B cells (CD19(+)), elevated the phagocytosis of PMN (p dioscorin and significant difference to those of the control (p dioscorin exhibited systemic and mucosal immunomodulatory activities after being ingested in vivo.

  15. New models for droplet heating and evaporation

    KAUST Repository

    Sazhin, Sergei S.; Elwardani, Ahmed Elsaid; Gusev, Ivan G.; Xie, Jianfei; Shishkova, Irina N.; Cao, Bingyang; Snegirev, Alexander Yu.; Heikal, Morgan Raymond

    2013-01-01

    and evaporation, taking into account the effects of the moving boundary due to evaporation, hydrodynamic models of multi-component droplet heating and evaporation, taking and not taking into account the effects of the moving boundary, new kinetic models of mono

  16. Droplet Manipulations in Two Phase Flow Microfluidics

    NARCIS (Netherlands)

    Pit, Arjen; Duits, Michael H.G.; Mugele, Friedrich Gunther

    2015-01-01

    Even though droplet microfluidics has been developed since the early 1980s, the number of applications that have resulted in commercial products is still relatively small. This is partly due to an ongoing maturation and integration of existing methods, but possibly also because of the emergence of

  17. Droplet microfluidic platform for cell electrofusion

    NARCIS (Netherlands)

    Schoeman, R.M.

    2015-01-01

    In this thesis a lab on a chip platform is described which is capable of electrofusing cells in a picoliter droplet. The platform consist out of glass part containing recessed platinum electrodes plasma bonded to a PDMS slab containing microchannels. First the two cell populations are introduced

  18. Droplet bubbling evaporatively cools a blowfly.

    Science.gov (United States)

    Gomes, Guilherme; Köberle, Roland; Von Zuben, Claudio J; Andrade, Denis V

    2018-04-19

    Terrestrial animals often use evaporative cooling to lower body temperature. Evaporation can occur from humid body surfaces or from fluids interfaced to the environment through a number of different mechanisms, such as sweating or panting. In Diptera, some flies move tidally a droplet of fluid out and then back in the buccopharyngeal cavity for a repeated number of cycles before eventually ingesting it. This is referred to as the bubbling behaviour. The droplet fluid consists of a mix of liquids from the ingested food, enzymes from the salivary glands, and antimicrobials, associated to the crop organ system, with evidence pointing to a role in liquid meal dehydration. Herein, we demonstrate that the bubbling behaviour also serves as an effective thermoregulatory mechanism to lower body temperature by means of evaporative cooling. In the blowfly, Chrysomya megacephala, infrared imaging revealed that as the droplet is extruded, evaporation lowers the fluid´s temperature, which, upon its re-ingestion, lowers the blowfly's body temperature. This effect is most prominent at the cephalic region, less in the thorax, and then in the abdomen. Bubbling frequency increases with ambient temperature, while its cooling efficiency decreases at high air humidities. Heat transfer calculations show that droplet cooling depends on a special heat-exchange dynamic, which result in the exponential activation of the cooling effect.

  19. Moving droplets : The measurement of contact lines

    NARCIS (Netherlands)

    Poelma, C.; Franken, M.J.Z.; Kim, H.; Westerweel, J.

    2014-01-01

    Contact lines are the locations where a gas, liquid and a solid meet. From everyday experience we know that such contact lines can be mobile, for example in the case of a water droplet sliding over a glass surface. However, the continuum description of the flow towards or away from a contact line

  20. Interaction between liquid droplets and heated surface

    Energy Technology Data Exchange (ETDEWEB)

    Nigmatulin, B I [Research and Engineering Centre, LWR Nuclear Plants Safety, Elektrogorsk (Russian Federation); Vasiliev, N I [Research and Engineering Centre, LWR Nuclear Plants Safety, Elektrogorsk (Russian Federation); Guguchkin, V V [Research and Engineering Centre, LWR Nuclear Plants Safety, Elektrogorsk (Russian Federation)

    1993-06-01

    In this paper, experimental methods and investigation results of interaction between droplets of different liquids and a heated surface are presented. Wetted area, contact time period and transition boundary from wetted to non-wetted interaction regimes are experimentally evaluated. A simple connection of the wetted area value and contact time period with the heat removal efficiency is shown. (orig.)

  1. Deformable nematic droplets in a magnetic field

    NARCIS (Netherlands)

    Otten, R.H.J.; van der Schoot, P. P. A. M.

    2012-01-01

    We present a Frank-Oseen elasticity theory for the shape and structure of deformable nematic droplets with homeotropic surface anchoring in the presence of a magnetic field. Inspired by recent experimental observations, we focus on the case where the magnetic susceptibility is negative, and find

  2. Adjuvants for single droplet application of glyphosate

    DEFF Research Database (Denmark)

    Mathiassen, Solvejg Kopp; Kudsk, Per; Lund, Ivar

    2016-01-01

    Retention and biological activity of droplets of glyphosate deposited onto plant leaves using a Drop on Demand inkjet printer application system, was examined on pot-grown Brassica napus, Solanum nigrum, Chenopodium album, Silene noctiflora and Echinocloa crus-galli plants. Retention was measured...

  3. Droplet-model electric dipole moments

    International Nuclear Information System (INIS)

    Myers, W.D.; Swiatecki, W.J.

    1991-01-01

    Denisov's recent criticism of the droplet-model formula for the dipole moment of a deformed nucleus as derived by Dorso et al., it shown to be invalid. This helps to clarify the relation of theory to the measured dipole moments, as discussed in the review article by Aberg et al. (orig.)

  4. Hydrodynamic clustering of droplets in turbulence

    Science.gov (United States)

    Kunnen, Rudie; Yavuz, Altug; van Heijst, Gertjan; Clercx, Herman

    2017-11-01

    Small, inertial particles are known to cluster in turbulent flows: particles are centrifuged out of eddies and gather in the strain-dominated regions. This so-called preferential concentration is reflected in the radial distribution function (RDF; a quantitative measure of clustering). We study clustering of water droplets in a loudspeaker-driven turbulence chamber. We track the motion of droplets in 3D and calculate the RDF. At moderate scales (a few Kolmogorov lengths) we find the typical power-law scaling of preferential concentration in the RDF. However, at even smaller scales (a few droplet diameters), we encounter a hitherto unobserved additional clustering. We postulate that the additional clustering is due to hydrodynamic interactions, an effect which is typically disregarded in modeling. Using a perturbative expansion of inertial effects in a Stokes-flow description of two interacting spheres, we obtain an expression for the RDF which indeed includes the additional clustering. The additional clustering enhances the collision probability of droplets, which enhances their growth rate due to coalescence. The additional clustering is thus an essential effect in precipitation modeling.

  5. Modeling Droplet Heat and Mass Transfer during Spray Bar Pressure Control of the Multipurpose Hydrogen Test Bed (MHTB) Tank in Normal Gravity

    Science.gov (United States)

    Kartuzova, O.; Kassemi, M.

    2016-01-01

    A CFD model for simulating pressure control in cryogenic storage tanks through the injection of a subcooled liquid into the ullage is presented and applied to the 1g MHTB spray bar cooling experiments. An Eulerian-Lagrangian approach is utilized to track the spray droplets and capture the interaction between the discrete droplets and the continuous ullage phase. The spray model is coupled with the VOF model by performing particle tracking in the ullage, removing particles from the ullage when they reach the interface, and then adding their contributions to the liquid. A new model for calculating the droplet-ullage heat and mass transfer is developed. In this model, a droplet is allowed to warm up to the saturation temperature corresponding to the ullage vapor pressure, after which it evaporates while remaining at the saturation temperature. The droplet model is validated against the results of the MHTB spray-bar cooling experiments with 50% and 90% tank fill ratios. The predictions of the present T-sat based model are compared with those of a previously developed kinetic-based droplet mass transfer model. The predictions of the two models regarding the evolving tank pressure and temperature distributions, as well as the droplets' trajectories and temperatures, are examined and compared in detail. Finally, the ullage pressure and local vapor and liquid temperature evolutions are validated against the corresponding data provided by the MHTB spray bar mixing experiment.

  6. Operator formulation of the droplet model

    International Nuclear Information System (INIS)

    Lee, B.W.

    1987-01-01

    We study in detail the implications of the operator formulation of the droplet model. The picture of high-energy scattering that emerges from this model attributed the interaction between two colliding particles at high energies to an instantaneous, multiple exchange between two extended charge distributions. Thus the study of charge correlation functions becomes the most important problem in the droplet model. We find that in order for the elastic cross section to have a finite limit at infinite energy, the charge must be a conserved one. In quantum electrodynamics the charge in question is the electric charge. In hadronic physics, we conjecture, it is the baryonic charge. Various arguments for and implications of this hypothesis are presented. We study formal properties of the charge correlation functions that follow from microcausality, T, C, P invariances, and charge conservation. Perturbation expansion of the correlation functions is studied, and their cluster properties are deduced. A cluster expansion of the high-energy T matrix is developed, and the exponentiation of the interaction potential in this scheme is noted. The operator droplet model is put to the test of reproducing the high-energy limit of elastic scattering quantum electrodynamics found by Cheng and Wu in perturbation theory. We find that the droplet model reproduces exactly the results of Cheng and Wu as to the impact factor. In fact, the ''impact picture'' of Cheng and Wu is completely equivalent to the droplet model in the operator version. An appraisal is made of the possible limitation of the model. (author). 13 refs

  7. Engineering plant membranes using droplet interface bilayers.

    Science.gov (United States)

    Barlow, N E; Smpokou, E; Friddin, M S; Macey, R; Gould, I R; Turnbull, C; Flemming, A J; Brooks, N J; Ces, O; Barter, L M C

    2017-03-01

    Droplet interface bilayers (DIBs) have become widely recognised as a robust platform for constructing model membranes and are emerging as a key technology for the bottom-up assembly of synthetic cell-like and tissue-like structures. DIBs are formed when lipid-monolayer coated water droplets are brought together inside a well of oil, which is excluded from the interface as the DIB forms. The unique features of the system, compared to traditional approaches (e.g., supported lipid bilayers, black lipid membranes, and liposomes), is the ability to engineer multi-layered bilayer networks by connecting multiple droplets together in 3D, and the capability to impart bilayer asymmetry freely within these droplet architectures by supplying droplets with different lipids. Yet despite these achievements, one potential limitation of the technology is that DIBs formed from biologically relevant components have not been well studied. This could limit the reach of the platform to biological systems where bilayer composition and asymmetry are understood to play a key role. Herein, we address this issue by reporting the assembly of asymmetric DIBs designed to replicate the plasma membrane compositions of three different plant species; Arabidopsis thaliana , tobacco, and oats, by engineering vesicles with different amounts of plant phospholipids, sterols and cerebrosides for the first time. We show that vesicles made from our plant lipid formulations are stable and can be used to assemble asymmetric plant DIBs. We verify this using a bilayer permeation assay, from which we extract values for absolute effective bilayer permeation and bilayer stability. Our results confirm that stable DIBs can be assembled from our plant membrane mimics and could lead to new approaches for assembling model systems to study membrane translocation and to screen new agrochemicals in plants.

  8. Chemotactic droplet swimmers in complex geometries

    Science.gov (United States)

    Jin, Chenyu; Hokmabad, Babak V.; Baldwin, Kyle A.; Maass, Corinna C.

    2018-02-01

    Chemotaxis1 and auto-chemotaxis are key mechanisms in the dynamics of micro-organisms, e.g. in the acquisition of nutrients and in the communication between individuals, influencing the collective behaviour. However, chemical signalling and the natural environment of biological swimmers are generally complex, making them hard to access analytically. We present a well-controlled, tunable artificial model to study chemotaxis and autochemotaxis in complex geometries, using microfluidic assays of self-propelling oil droplets in an aqueous surfactant solution (Herminghaus et al 2014 Soft Matter 10 7008-22 Krüger et al 2016 Phys. Rev. Lett. 117). Droplets propel via interfacial Marangoni stresses powered by micellar solubilisation. Moreover, filled micelles act as a chemical repellent by diffusive phoretic gradient forces. We have studied these chemotactic effects in a series of microfluidic geometries, as published in Jin et al (2017 Proc. Natl Acad. Sci. 114 5089-94): first, droplets are guided along the shortest path through a maze by surfactant diffusing into the maze from the exit. Second, we let auto-chemotactic droplet swimmers pass through bifurcating microfluidic channels and record anticorrelations between the branch choices of consecutive droplets. We present an analytical Langevin model matching the experimental data. In a previously unpublished experiment, pillar arrays of variable sizes and shapes provide a convex wall interacting with the swimmer and, in the case of attachment, bending its trajectory and forcing it to revert to its own trail. We observe different behaviours based on the interplay of wall curvature and negative autochemotaxis, i.e. no attachment for highly curved interfaces, stable trapping at large pillars, and a narrow transition region where negative autochemotaxis makes the swimmers detach after a single orbit.

  9. An interface tracking model for droplet electrocoalescence.

    Energy Technology Data Exchange (ETDEWEB)

    Erickson, Lindsay Crowl

    2013-09-01

    This report describes an Early Career Laboratory Directed Research and Development (LDRD) project to develop an interface tracking model for droplet electrocoalescence. Many fluid-based technologies rely on electrical fields to control the motion of droplets, e.g. microfluidic devices for high-speed droplet sorting, solution separation for chemical detectors, and purification of biodiesel fuel. Precise control over droplets is crucial to these applications. However, electric fields can induce complex and unpredictable fluid dynamics. Recent experiments (Ristenpart et al. 2009) have demonstrated that oppositely charged droplets bounce rather than coalesce in the presence of strong electric fields. A transient aqueous bridge forms between approaching drops prior to pinch-off. This observation applies to many types of fluids, but neither theory nor experiments have been able to offer a satisfactory explanation. Analytic hydrodynamic approximations for interfaces become invalid near coalescence, and therefore detailed numerical simulations are necessary. This is a computationally challenging problem that involves tracking a moving interface and solving complex multi-physics and multi-scale dynamics, which are beyond the capabilities of most state-of-the-art simulations. An interface-tracking model for electro-coalescence can provide a new perspective to a variety of applications in which interfacial physics are coupled with electrodynamics, including electro-osmosis, fabrication of microelectronics, fuel atomization, oil dehydration, nuclear waste reprocessing and solution separation for chemical detectors. We present a conformal decomposition finite element (CDFEM) interface-tracking method for the electrohydrodynamics of two-phase flow to demonstrate electro-coalescence. CDFEM is a sharp interface method that decomposes elements along fluid-fluid boundaries and uses a level set function to represent the interface.

  10. Energy storage

    International Nuclear Information System (INIS)

    2012-01-01

    After having outlined the importance of energy storage in the present context, this document outlines that it is an answer to economic, environmental and technological issues. It proposes a brief overview of the various techniques of energy storage: under the form of chemical energy (hydrocarbons, biomass, hydrogen production), thermal energy (sensitive or latent heat storage), mechanical energy (potential energy by hydraulic or compressed air storage, kinetic energy with flywheels), electrochemical energy (in batteries), electric energy (super-capacitors, superconductor magnetic energy storage). Perspectives are briefly evoked

  11. Stability and in vitro digestibility of emulsions containing lecithin and whey proteins.

    Science.gov (United States)

    Mantovani, Raphaela Araujo; Cavallieri, Ângelo Luiz Fazani; Netto, Flavia Maria; Cunha, Rosiane Lopes

    2013-09-01

    The effect of pH and high-pressure homogenization on the properties of oil-in-water (O/W) emulsions stabilized by lecithin and/or whey proteins (WPI) was evaluated. For this purpose, emulsions were characterized by visual analysis, droplet size distribution, zeta potential, electrophoresis, rheological measurements and their response to in vitro digestion. Lecithin emulsions were stable even after 7 days of storage and WPI emulsions were unstable only at pH values close to the isoelectric point (pI) of proteins. Systems containing the mixture of lecithin and WPI showed high kinetic instability at pH 3, which was attributed to the electrostatic interaction between the emulsifiers oppositely charged at this pH value. At pH 5.5 and 7, the mixture led to reduction of the droplet size with enhanced emulsion stability compared to the systems with WPI or lecithin. The stability of WPI emulsions after the addition of lecithin, especially at pH 5.5, was associated with the increase of droplet surface charge density. The in vitro digestion evaluation showed that WPI emulsion was more stable against gastrointestinal conditions.

  12. Droplet Epitaxy Image Contrast in Mirror Electron Microscopy

    Science.gov (United States)

    Kennedy, S. M.; Zheng, C. X.; Jesson, D. E.

    2017-01-01

    Image simulation methods are applied to interpret mirror electron microscopy (MEM) images obtained from a movie of GaAs droplet epitaxy. Cylindrical symmetry of structures grown by droplet epitaxy is assumed in the simulations which reproduce the main features of the experimental MEM image contrast, demonstrating that droplet epitaxy can be studied in real-time. It is therefore confirmed that an inner ring forms at the droplet contact line and an outer ring (or skirt) occurs outside the droplet periphery. We believe that MEM combined with image simulations will be increasingly used to study the formation and growth of quantum structures.

  13. Hydrodynamics of Leidenfrost droplets in one-component fluids

    KAUST Repository

    Xu, Xinpeng; Qian, Tiezheng

    2013-01-01

    Using the dynamic van der Waals theory [Phys. Rev. E 75, 036304 (2007)], we numerically investigate the hydrodynamics of Leidenfrost droplets under gravity in two dimensions. Some recent theoretical predictions and experimental observations are confirmed in our simulations. A Leidenfrost droplet larger than a critical size is shown to be unstable and break up into smaller droplets due to the Rayleigh-Taylor instability of the bottom surface of the droplet. Our simulations demonstrate that an evaporating Leidenfrost droplet changes continuously from a puddle to a circular droplet, with the droplet shape controlled by its size in comparison with a few characteristic length scales. The geometry of the vapor layer under the droplet is found to mainly depend on the droplet size and is nearly independent of the substrate temperature, as reported in a recent experimental study [Phys. Rev. Lett. 109, 074301 (2012)]. Finally, our simulations demonstrate that a Leidenfrost droplet smaller than a characteristic size takes off from the hot substrate because the levitating force due to evaporation can no longer be balanced by the weight of the droplet, as observed in a recent experimental study [Phys. Rev. Lett. 109, 034501 (2012)].

  14. Hydrodynamics of Leidenfrost droplets in one-component fluids

    KAUST Repository

    Xu, Xinpeng

    2013-04-24

    Using the dynamic van der Waals theory [Phys. Rev. E 75, 036304 (2007)], we numerically investigate the hydrodynamics of Leidenfrost droplets under gravity in two dimensions. Some recent theoretical predictions and experimental observations are confirmed in our simulations. A Leidenfrost droplet larger than a critical size is shown to be unstable and break up into smaller droplets due to the Rayleigh-Taylor instability of the bottom surface of the droplet. Our simulations demonstrate that an evaporating Leidenfrost droplet changes continuously from a puddle to a circular droplet, with the droplet shape controlled by its size in comparison with a few characteristic length scales. The geometry of the vapor layer under the droplet is found to mainly depend on the droplet size and is nearly independent of the substrate temperature, as reported in a recent experimental study [Phys. Rev. Lett. 109, 074301 (2012)]. Finally, our simulations demonstrate that a Leidenfrost droplet smaller than a characteristic size takes off from the hot substrate because the levitating force due to evaporation can no longer be balanced by the weight of the droplet, as observed in a recent experimental study [Phys. Rev. Lett. 109, 034501 (2012)].

  15. Binary particle separation in droplet microfluidics using acoustophoresis

    Science.gov (United States)

    Fornell, Anna; Cushing, Kevin; Nilsson, Johan; Tenje, Maria

    2018-02-01

    We show a method for separation of two particle species with different acoustic contrasts originally encapsulated in the same droplet in a continuous two-phase system. This was realized by using bulk acoustic standing waves in a 380 μm wide silicon-glass microfluidic channel. Polystyrene particles (positive acoustic contrast particles) and in-house synthesized polydimethylsiloxane (PDMS) particles (negative acoustic contrast particles) were encapsulated inside water-in-oil droplets either individually or in a mixture. At acoustic actuation of the system at the fundamental resonance frequency, the polystyrene particles were moved to the center of the droplet (pressure node), while the PDMS particles were moved to the sides of the droplet (pressure anti-nodes). The acoustic particle manipulation step was combined in series with a trifurcation droplet splitter, and as the original droplet passed through the splitter and was divided into three daughter droplets, the polystyrene particles were directed into the center daughter droplet, while the PDMS particles were directed into the two side daughter droplets. The presented method expands the droplet microfluidics tool-box and offers new possibilities to perform binary particle separation in droplet microfluidic systems.

  16. Evaluation of droplet size distributions using univariate and multivariate approaches

    DEFF Research Database (Denmark)

    Gauno, M.H.; Larsen, C.C.; Vilhelmsen, T.

    2013-01-01

    of the distribution. The current study was aiming to compare univariate and multivariate approach in evaluating droplet size distributions. As a model system, the atomization of a coating solution from a two-fluid nozzle was investigated. The effect of three process parameters (concentration of ethyl cellulose...... in ethanol, atomizing air pressure, and flow rate of coating solution) on the droplet size and droplet size distribution using a full mixed factorial design was used. The droplet size produced by a two-fluid nozzle was measured by laser diffraction and reported as volume based size distribution....... Investigation of loading and score plots from principal component analysis (PCA) revealed additional information on the droplet size distributions and it was possible to identify univariate statistics (volume median droplet size), which were similar, however, originating from varying droplet size distributions...

  17. Supersonic laser-induced jetting of aluminum micro-droplets

    Energy Technology Data Exchange (ETDEWEB)

    Zenou, M. [Racah Institute of Physics and the Harvey M. Kruger Family Center for Nano-science and Nanotechnology, The Hebrew University of Jerusalem, 91904 Jerusalem (Israel); Additive Manufacturing Lab, Orbotech Ltd., P.O. Box 215, 81101 Yavne (Israel); Sa' ar, A. [Racah Institute of Physics and the Harvey M. Kruger Family Center for Nano-science and Nanotechnology, The Hebrew University of Jerusalem, 91904 Jerusalem (Israel); Kotler, Z. [Additive Manufacturing Lab, Orbotech Ltd., P.O. Box 215, 81101 Yavne (Israel)

    2015-05-04

    The droplet velocity and the incubation time of pure aluminum micro-droplets, printed using the method of sub-nanosecond laser induced forward transfer, have been measured indicating the formation of supersonic laser-induced jetting. The incubation time and the droplet velocity were extracted by measuring a transient electrical signal associated with droplet landing on the surface of the acceptor substrate. This technique has been exploited for studying small volume droplets, in the range of 10–100 femto-litters for which supersonic velocities were measured. The results suggest elastic propagation of the droplets across the donor-to-acceptor gap, a nonlinear deposition dynamics on the surface of the acceptor and overall efficient energy transfer from the laser beam to the droplets.

  18. Supersonic laser-induced jetting of aluminum micro-droplets

    International Nuclear Information System (INIS)

    Zenou, M.; Sa'ar, A.; Kotler, Z.

    2015-01-01

    The droplet velocity and the incubation time of pure aluminum micro-droplets, printed using the method of sub-nanosecond laser induced forward transfer, have been measured indicating the formation of supersonic laser-induced jetting. The incubation time and the droplet velocity were extracted by measuring a transient electrical signal associated with droplet landing on the surface of the acceptor substrate. This technique has been exploited for studying small volume droplets, in the range of 10–100 femto-litters for which supersonic velocities were measured. The results suggest elastic propagation of the droplets across the donor-to-acceptor gap, a nonlinear deposition dynamics on the surface of the acceptor and overall efficient energy transfer from the laser beam to the droplets

  19. Symptomatic lipid storage in carriers for the PNPLA2 gene

    NARCIS (Netherlands)

    Janssen, M.C.H.; Engelen, B.G.M. van; Kapusta, L.; Lammens, M.M.; Dijk, M.; Fischer, J.; Graaf, M. van der; Wevers, R.A.; Fahrleitner, M.; Zimmermann, R.; Morava, E.

    2013-01-01

    Neutral lipid storage disease comprises a heterogeneous group of inherited disorders characterized by severe accumulation of cytoplasmic triglyceride droplets in several tissues and neutrophils. A novel type of autosomal recessive lipid myopathy due to PNPLA2 mutations was recently described with

  20. Global rainbow refractometry for droplet temperature measurement

    International Nuclear Information System (INIS)

    Pascal Lemaitre; Emmanuel Porcheron; Amandine Nuboer; Philippe Brun; Pierre Cornet; Jeanne Malet; Jacques Vendel; Laurent Bouilloux; Gerard Grehan

    2005-01-01

    Full text of publication follows: In order to establish an accurate database to characterize the heat and mass transfers between a spray and the atmosphere with thermal-hydraulic conditions representative of a hypothetical nuclear reactor accident in the containment enclosure of a pressurized water reactor (PWR), the French Institut de Radioprotection et de Surete Nucleaire (IRSN) has developed the TOSQAN experimental facility. This experiment is highly instrumented with non-intrusive diagnostics allowing to measure droplet size and velocity and gas concentrations [1]. The aim of this work is to present the Global Rainbow Thermometry (GRT), which is an advanced non-intrusive optical diagnostic, developed to measure the mean temperature of a set of falling droplets, in a measurement volume of 1 cm 3 . The final paper will be divided in three parts. In the first one, we will explain the principle of the rainbow formation and how droplet temperature can be deduced from the rainbow analysis [2]. This part will be illustrated with the theoretical background on the rainbow and numerical simulations of the global rainbow. The second part will be devoted to present the global rainbow experimental set-up we have developed on optical table, its experimental qualification and finally its implementation on the TOSQAN facility [3]. Finally, we will present the temperature measurements achieved in TOSQAN for thermal-hydraulic conditions representative of a hypothetical nuclear reactor accident. These measurements are useful to characterize the heat and mass transfers between the spraying droplets and the air-steam mixture composing the atmosphere. This analysis will be exposed in a two companion papers. References: [1] E. Porcheron, P. Brun, P. Cornet, J. Malet, J. Vendel. Optical diagnostics applied for single and multi-phase flow characterization in the TOSQAN facility dedicated for thermal hydraulic containment studies. NURETH-10 Seoul, Korea, October 5-9, 2003. [2] P

  1. Global rainbow refractometry for droplet temperature measurement

    Energy Technology Data Exchange (ETDEWEB)

    Pascal Lemaitre; Emmanuel Porcheron; Amandine Nuboer; Philippe Brun; Pierre Cornet; Jeanne Malet; Jacques Vendel; Laurent Bouilloux [Institut de Radioprotection et de Surete Nucleaire DSU/SERAC, BP 68, 91192 Gif-sur-Yvette Cedex (France); Gerard Grehan [UMR 6614 CORIA, Laboratoire d' Electromagnetisme et Systemes Particulaires Site Universitaire du Madrillet, Avenue de l' universite BP 12, 76 801 Saint Etienne du Rouvray Cedex, (France)

    2005-07-01

    Full text of publication follows: In order to establish an accurate database to characterize the heat and mass transfers between a spray and the atmosphere with thermal-hydraulic conditions representative of a hypothetical nuclear reactor accident in the containment enclosure of a pressurized water reactor (PWR), the French Institut de Radioprotection et de Surete Nucleaire (IRSN) has developed the TOSQAN experimental facility. This experiment is highly instrumented with non-intrusive diagnostics allowing to measure droplet size and velocity and gas concentrations [1]. The aim of this work is to present the Global Rainbow Thermometry (GRT), which is an advanced non-intrusive optical diagnostic, developed to measure the mean temperature of a set of falling droplets, in a measurement volume of 1 cm{sup 3}. The final paper will be divided in three parts. In the first one, we will explain the principle of the rainbow formation and how droplet temperature can be deduced from the rainbow analysis [2]. This part will be illustrated with the theoretical background on the rainbow and numerical simulations of the global rainbow. The second part will be devoted to present the global rainbow experimental set-up we have developed on optical table, its experimental qualification and finally its implementation on the TOSQAN facility [3]. Finally, we will present the temperature measurements achieved in TOSQAN for thermal-hydraulic conditions representative of a hypothetical nuclear reactor accident. These measurements are useful to characterize the heat and mass transfers between the spraying droplets and the air-steam mixture composing the atmosphere. This analysis will be exposed in a two companion papers. References: [1] E. Porcheron, P. Brun, P. Cornet, J. Malet, J. Vendel. Optical diagnostics applied for single and multi-phase flow characterization in the TOSQAN facility dedicated for thermal hydraulic containment studies. NURETH-10 Seoul, Korea, October 5-9, 2003. [2] P

  2. An experimental study on suspended sodium droplet combustion (3)

    International Nuclear Information System (INIS)

    Sato, Kenji

    2005-03-01

    As part of studies for phenomenological investigation of sodium droplet burning behavior, in our previous experimental studies for suspended single sodium droplet, behavior of ignition process and succeeding combustion, ignition delay time, and droplet temperature history had been investigated. In this study, combustion experiments of suspended sodium droplet were performed in upward dry air flow by expanding the range of free-stream velocity U of air flow into 400 cm/s with initial droplet temperature Ti=300, 350, and 400degC and initial droplet diameter 4 mm at first. Then, the combustion experiments were also performed by changing the initial droplet diameter from 2.3 to 4.4 mm with Ti=350 and 400degC and U=100 cm/s. From the experimental results, the effects of free-stream velocity, initial droplet temperature, and initial droplet diameter on the ignition/burning behavior and ignition delay time were examined. The obtained results are as follows: (1) Ignition phenomena of suspended droplet were observed for all examined experimental conditions up to 400 cm/s. The orange emission observed at the moment of ignition occurs simultaneously over whole droplet surface except the top region of it. (2) The feature of the dependence of ignition delay time on the free-stream velocity is independent of the initial droplet temperature. With the increase of the free-stream velocity, up to 300 cm/s the ignition delay time decreases with decreasing dependency, and then the dependency increases more. (3) The ignition delay time increases with the increase of initial droplet diameter. The dependency increases as the initial droplet diameter increases. The ignition delay time extrapolated toward zero diameters from the obtained results becomes to be essentially zero. (author)

  3. Protein replacement therapy partially corrects the cholesterol-storage phenotype in a mouse model of Niemann-Pick type C2 disease.

    Directory of Open Access Journals (Sweden)

    Gitte Krogh Nielsen

    Full Text Available Niemann-Pick type C2 (NPC2 disease is a fatal autosomal recessive neurovisceral degenerative disorder characterized by late endosomal-lysosomal sequestration of low-density lipoprotein derived cholesterol. The breach in intracellular cholesterol homeostasis is caused by deficiency of functional NPC2, a soluble sterol binding protein targeted to the lysosomes by binding the mannose-6-phosphate receptor. As currently there is no effective treatment for the disorder, we have investigated the efficacy of NPC2 replacement therapy in a murine gene-trap model of NPC2-disease generated on the 129P2/OlaHsd genetic background. NPC2 was purified from bovine milk and its functional competence assured in NPC2-deficient fibroblasts using the specific cholesterol fluorescent probe filipin. For evaluation of phenotype correction in vivo, three-week-old NPC2(-/- mice received two weekly intravenous injections of 5 mg/kg NPC2 until trial termination 66 days later. Whereas the saline treated NPC2(-/- mice exhibited massive visceral cholesterol storage as compared to their wild-type littermates, administration of NPC2 caused a marked reduction in cholesterol build up. The histological findings, indicating an amelioration of the disease pathology in liver, spleen, and lungs, corroborated the biochemical results. Little or no difference in the overall cholesterol levels was observed in the kidneys, blood, cerebral cortex and hippocampus when comparing NPC2(-/- and wild type mice. However, cerebellum cholesterol was increased about two fold in NPC2(-/- mice compared with wild-type littermates. Weight gain performance was slightly improved as a result of the NPC2 treatment but significant motor coordination deficits were still observed. Accordingly, ultrastructural cerebellar abnormalities were detected in both saline treated and NPC2 treated NPC2(-/- animals 87 days post partum. Our data indicate that protein replacement may be a beneficial therapeutic approach in the

  4. Laser-induced fluorescence imaging of acetone inside evaporating and burning fuel droplets

    Science.gov (United States)

    Shringi, D. S.; Shaw, B. D.; Dwyer, H. A.

    2009-01-01

    Laser-induced fluorescence was used to visualize acetone fields inside individual droplets of pure acetone as well as droplets composed of methanol or 1-propanol initially mixed with acetone. Droplets were supported on a horizontal wire and two vaporization conditions were investigated: (1) slow evaporation in room air and (2) droplet combustion, which leads to substantially faster droplet surface regression rates. Acetone was preferentially gasified, causing its concentration in droplets to drop in time with resultant decreases in acetone fluorescence intensities. Slowly vaporizing droplets did not exhibit large spatial variations of fluorescence within droplets, indicating that these droplets were relatively well mixed. Ignition of droplets led to significant variations in fluorescence intensities within droplets, indicating that these droplets were not well mixed. Ignited droplets composed of mixtures of 1-propanol and acetone showed large time-varying changes in shapes for higher acetone concentrations, suggesting that bubble formation was occurring in these droplets.

  5. Distinct populations of hepatic stellate cells in the mouse liver have different capacities for retinoid and lipid storage.

    Directory of Open Access Journals (Sweden)

    Diana N D'Ambrosio

    Full Text Available Hepatic stellate cell (HSC lipid droplets are specialized organelles for the storage of retinoid, accounting for 50-60% of all retinoid present in the body. When HSCs activate, retinyl ester levels progressively decrease and the lipid droplets are lost. The objective of this study was to determine if the HSC population in a healthy, uninjured liver demonstrates heterogeneity in its capacity for retinoid and lipid storage in lipid droplets. To this end, we utilized two methods of HSC isolation, which leverage distinct properties of these cells, including their vitamin A content and collagen expression. HSCs were isolated either from wild type (WT mice in the C57BL/6 genetic background by flotation in a Nycodenz density gradient, followed by fluorescence activated cell sorting (FACS based on vitamin A autofluorescence, or from collagen-green fluorescent protein (GFP mice by FACS based on GFP expression from a GFP transgene driven by the collagen I promoter. We show that GFP-HSCs have: (i increased expression of typical markers of HSC activation; (ii decreased retinyl ester levels, accompanied by reduced expression of the enzyme needed for hepatic retinyl ester synthesis (LRAT; (iii decreased triglyceride levels; (iv increased expression of genes associated with lipid catabolism; and (v an increase in expression of the retinoid-catabolizing cytochrome, CYP2S1.Our observations suggest that the HSC population in a healthy, uninjured liver is heterogeneous. One subset of the total HSC population, which expresses early markers of HSC activation, may be "primed" and ready for rapid response to acute liver injury.

  6. Mechanism of supercooled droplet freezing on surfaces.

    Science.gov (United States)

    Jung, Stefan; Tiwari, Manish K; Doan, N Vuong; Poulikakos, Dimos

    2012-01-10

    Understanding ice formation from supercooled water on surfaces is a problem of fundamental importance and general utility. Superhydrophobic surfaces promise to have remarkable 'icephobicity' and low ice adhesion. Here we show that their icephobicity can be rendered ineffective by simple changes in environmental conditions. Through experiments, nucleation theory and heat transfer physics, we establish that humidity and/or the flow of a surrounding gas can fundamentally switch the ice crystallization mechanism, drastically affecting surface icephobicity. Evaporative cooling of the supercooled liquid can engender ice crystallization by homogeneous nucleation at the droplet-free surface as opposed to the expected heterogeneous nucleation at the substrate. The related interplay between droplet roll-off and rapid crystallization is also studied. Overall, we bring a novel perspective to icing and icephobicity, unveiling the strong influence of environmental conditions in addition to the accepted effects of the surface conditions and hydrophobicity.

  7. Magnetic droplet soliton nucleation in oblique fields

    Science.gov (United States)

    Mohseni, Morteza; Hamdi, M.; Yazdi, H. F.; Banuazizi, S. A. H.; Chung, S.; Sani, S. R.; Åkerman, Johan; Mohseni, Majid

    2018-05-01

    We study the auto-oscillating magnetodynamics in orthogonal spin-torque nano-oscillators (STNOs) as a function of the out-of-plane (OOP) magnetic-field angle. In perpendicular fields and at OOP field angles down to approximately 50°, we observe the nucleation of a droplet. However, for field angles below 50°, experiments indicate that the droplet gives way to propagating spin waves, in agreement with our micromagnetic simulations. Theoretical calculations show that the physical mechanism behind these observations is the sign changing of spin-wave nonlinearity (SWN) by angle. In addition, we show that the presence of a strong perpendicular magnetic anisotropy free layer in the system reverses the angular dependence of the SWN and dynamics in STNOs with respect to the known behavior determined for the in-plane magnetic anisotropy free layer. Our results are of fundamental interest in understanding the rich dynamics of nanoscale solitons and spin-wave dynamics in STNOs.

  8. Ballistic model to estimate microsprinkler droplet distribution

    Directory of Open Access Journals (Sweden)

    Conceição Marco Antônio Fonseca

    2003-01-01

    Full Text Available Experimental determination of microsprinkler droplets is difficult and time-consuming. This determination, however, could be achieved using ballistic models. The present study aimed to compare simulated and measured values of microsprinkler droplet diameters. Experimental measurements were made using the flour method, and simulations using a ballistic model adopted by the SIRIAS computational software. Drop diameters quantified in the experiment varied between 0.30 mm and 1.30 mm, while the simulated between 0.28 mm and 1.06 mm. The greatest differences between simulated and measured values were registered at the highest radial distance from the emitter. The model presented a performance classified as excellent for simulating microsprinkler drop distribution.

  9. Swimming droplets driven by a surface wave

    Science.gov (United States)

    Ebata, Hiroyuki; Sano, Masaki

    2015-02-01

    Self-propelling motion is ubiquitous for soft active objects such as crawling cells, active filaments, and liquid droplets moving on surfaces. Deformation and energy dissipation are required for self-propulsion of both living and non-living matter. From the perspective of physics, searching for universal laws of self-propelled motions in a dissipative environment is worthwhile, regardless of the objects' details. In this article, we propose a simple experimental system that demonstrates spontaneous migration of a droplet under uniform mechanical agitation. As we vary control parameters, spontaneous symmetry breaking occurs sequentially, and cascades of bifurcations of the motion arise. Equations describing deformable particles and hydrodynamic simulations successfully describe all of the observed motions. This system should enable us to improve our understanding of spontaneous motions of self-propelled objects.

  10. Lipid Structure in Triolein Lipid Droplets

    DEFF Research Database (Denmark)

    Chaban, Vitaly V; Khandelia, Himanshu

    2014-01-01

    of a mass of hydrophobic lipid esters coved by phospholipid monolayer. The small size and unique architecture of LDs makes it complicated to study LD structure by modern experimental methods. We discuss coarse-grained molecular dynamics (MD) simulations of LD formation in systems containing 1-palmitoyl-2...... to coarse-grained simulations, the presence of PE lipids at the interface has a little impact on distribution of components and on the overall LD structure. (4) The thickness of the lipid monolayer at the surface of the droplet is similar to the thickness of one leaflet of a bilayer. Computer simulations......Lipid droplets (LDs) are primary repositories of esterified fatty acids and sterols in animal cells. These organelles originate on the lumenal or cytoplasmic side of endoplasmic reticulum (ER) membrane and are released to the cytosol. In contrast to other intracellular organelles, LDs are composed...

  11. Neutron storage

    International Nuclear Information System (INIS)

    Strelkov, A.V.

    2004-01-01

    The report is devoted to neutron storage (NS) and describes the history of experiments on the NS development. Great attention is paid to ultracold neutron (UCN) storage. The experiments on the UCN generation, transport, spectroscopy, storage and detection are described. Experiments on searching the UCN electric-dipole moment and electric charge are continued. Possible using of UCN for studying the nanoparticles is discussed [ru

  12. Energy storage

    International Nuclear Information System (INIS)

    Odru, P.

    2010-01-01

    This book proposes a broad overview of the technologies developed in the domains of on-board electricity storage (batteries, super-capacitors, flywheels), stationary storage (hydraulic dams, compressed air, batteries and hydrogen), and heat storage (sensible, latent and sorption) together with their relative efficiency, their expected developments and what advantages they can offer. Eminent specialists of this domain have participated to the redaction of this book, all being members of the Tuck's Foundation 'IDees' think tank. (J.S.)

  13. Energy storage

    CERN Document Server

    Brunet, Yves

    2013-01-01

    Energy storage examines different applications such as electric power generation, transmission and distribution systems, pulsed systems, transportation, buildings and mobile applications. For each of these applications, proper energy storage technologies are foreseen, with their advantages, disadvantages and limits. As electricity cannot be stored cheaply in large quantities, energy has to be stored in another form (chemical, thermal, electromagnetic, mechanical) and then converted back into electric power and/or energy using conversion systems. Most of the storage technologies are examined: b

  14. Tritium storage

    International Nuclear Information System (INIS)

    Hircq, B.

    1989-01-01

    A general synthesis about tritium storage is achieved in this paper and a particular attention is given to practical application in the Fusion Technology Program. Tritium, storage under gaseous form and solid form are discussed (characteristics, advantages, disadvantages and equipments). The way of tritium storage is then discussed and a choice established as a function of a logic which takes into account the main working parameters

  15. Lithography-free nanofluidic concentrator based on droplets-on-demand system

    Science.gov (United States)

    Yu, Miao; Zhou, Hongbo; Yao, Shuhuai

    2013-11-01

    Biomarkers are usually low-abundance proteins in biofluids and below detection limit of conventional biosensors. Nanofluidic concentration devices allow efficient biomolecules trapping by utilizing ion concentration polarization near nanochannels. However, once the electric field is turned off, the electrokinetic concentration plug cannot maintain its concentration status and starts to diffuse. In order to maintain the high concentration and extract the concentrated sample for further analysis, a good approach is to encapsulate these plugs into water-in-oil droplets. Here we developed a nanofluidic concentrator based on droplet-on-demand generator to encapsulate concentrated sample in nL droplets. The lithography-free nanochannels were patterned by thermal cracking on the surface of PS Petri-dish. The resulting nanochannel arrays were 30 nm in depth. In combination with microchannels on PDMS, the micro-nano hybrid chip was developed. We used FITC solution to demonstrate that the chip significantly increased the sample concentration for more than 100 folds within 5 minutes. By tuning the pulsed pressure imposed by the solenoid valve connected to the concentration channel, the system can generate a desired volume of droplet with a target sample concentration at a prescribed time. This work was supported by the Research Grants Council of Hong Kong under General Research Fund (Grant No. 621110).

  16. Droplet-model predictions of charge moments

    International Nuclear Information System (INIS)

    Myers, W.D.

    1982-04-01

    The Droplet Model expressions for calculating various moments of the nuclear charge distribution are given. There are contributions to the moments from the size and shape of the system, from the internal redistribution induced by the Coulomb repulsion, and from the diffuseness of the surface. A case is made for the use of diffuse charge distributions generated by convolution as an alternative to Fermi-functions

  17. Distribution of droplet sizes for seed solution

    International Nuclear Information System (INIS)

    Marwah, R.K.; Dixit, N.S.; Venkataramani, N.; Rohatgi, V.K.

    In open cycle MHD power generation, power is generated by passing seeded hot combustion products of a fossil fuel through a magnetic field. Seeding is done with a salt which is readily ionizable, preferably in the form of an aqueous solution, such as potassium carbonate, potassium sulphate, etc. Methods of atomization and the theoretical drop size calculations are presented. Basic parameters necessary for droplet size determination and their measurement are also described. (K.B.)

  18. Development of an imaging system for single droplet characterization using a droplet generator.

    Science.gov (United States)

    Minov, S Vulgarakis; Cointault, F; Vangeyte, J; Pieters, J G; Hijazi, B; Nuyttens, D

    2012-01-01

    The spray droplets generated by agricultural nozzles play an important role in the application accuracy and efficiency of plant protection products. The limitations of the non-imaging techniques and the recent improvements in digital image acquisition and processing increased the interest in using high speed imaging techniques in pesticide spray characterisation. The goal of this study was to develop an imaging technique to evaluate the characteristics of a single spray droplet using a piezoelectric single droplet generator and a high speed imaging technique. Tests were done with different camera settings, lenses, diffusers and light sources. The experiments have shown the necessity for having a good image acquisition and processing system. Image analysis results contributed in selecting the optimal set-up for measuring droplet size and velocity which consisted of a high speed camera with a 6 micros exposure time, a microscope lens at a working distance of 43 cm resulting in a field of view of 1.0 cm x 0.8 cm and a Xenon light source without diffuser used as a backlight. For measuring macro-spray characteristics as the droplet trajectory, the spray angle and the spray shape, a Macro Video Zoom lens at a working distance of 14.3 cm with a bigger field of view of 7.5 cm x 9.5 cm in combination with a halogen spotlight with a diffuser and the high speed camera can be used.

  19. Characteristics of droplet motion in effervescent sprays

    Directory of Open Access Journals (Sweden)

    Jedelský Jan

    2014-03-01

    Full Text Available Time resolved droplet size and velocity measurement was made using Phase-Doppler anemometry in an effervescent spray at GLR of 6 % and operation pressure drops 21 – 52 kPa. The spray shows a size dependent variation of mean as well as fluctuating axial and radial velocities of droplets similarly for all operation regimes. Particles under 13 μm follow the gas flow, axially decelerated due to gas expansion. Velocity of medium sized particles is positively size correlated and larger particles keep high velocity, given them during discharge. Fluctuating radial velocity of small particles is larger than that of large particles while fluctuating axial velocity increases with size. Small particles thus reach a ratio of radial to axial velocity fluctuations ~ 0.6 but large particles only ~ 0.1, which indicates large transverse dispersion of small particles. Overall fluctuating velocity ratios smaller than 0.5 document an anisotropic character of the liquid mass fluctuations. Power spectral density (PSD of axial velocity fluctuations of large droplets is uniform up to 1 kHz, while PSD of smaller particles drops down with frequency for frequencies > 100 Hz. Large particles thus preserve the fluctuations imposed during discharge while the gas turbulence drops with frequency. Turbulence intensity reaches 14 to 21 % depending on pressure. Such high-turbulence character of the flow probably results from a heterogeneous gas–liquid mixture at the discharge.

  20. Characteristics of droplet motion in effervescent sprays

    Science.gov (United States)

    Jedelský, Jan; Zaremba, Matouš; Malý, Milan; Jícha, Miroslav

    2014-03-01

    Time resolved droplet size and velocity measurement was made using Phase-Doppler anemometry in an effervescent spray at GLR of 6 % and operation pressure drops 21 - 52 kPa. The spray shows a size dependent variation of mean as well as fluctuating axial and radial velocities of droplets similarly for all operation regimes. Particles under 13 μm follow the gas flow, axially decelerated due to gas expansion. Velocity of medium sized particles is positively size correlated and larger particles keep high velocity, given them during discharge. Fluctuating radial velocity of small particles is larger than that of large particles while fluctuating axial velocity increases with size. Small particles thus reach a ratio of radial to axial velocity fluctuations ~ 0.6 but large particles only ~ 0.1, which indicates large transverse dispersion of small particles. Overall fluctuating velocity ratios smaller than 0.5 document an anisotropic character of the liquid mass fluctuations. Power spectral density (PSD) of axial velocity fluctuations of large droplets is uniform up to 1 kHz, while PSD of smaller particles drops down with frequency for frequencies > 100 Hz. Large particles thus preserve the fluctuations imposed during discharge while the gas turbulence drops with frequency. Turbulence intensity reaches 14 to 21 % depending on pressure. Such high-turbulence character of the flow probably results from a heterogeneous gas-liquid mixture at the discharge.

  1. Water droplet evaporation from sticky superhydrophobic surfaces

    Science.gov (United States)

    Lee, Moonchan; Kim, Wuseok; Lee, Sanghee; Baek, Seunghyeon; Yong, Kijung; Jeon, Sangmin

    2017-07-01

    The evaporation dynamics of water from sticky superhydrophobic surfaces was investigated using a quartz crystal microresonator and an optical microscope. Anodic aluminum oxide (AAO) layers with different pore sizes were directly fabricated onto quartz crystal substrates and hydrophobized via chemical modification. The resulting AAO layers exhibited hydrophobic or superhydrophobic characteristics with strong adhesion to water due to the presence of sealed air pockets inside the nanopores. After placing a water droplet on the AAO membranes, variations in the resonance frequency and Q-factor were measured throughout the evaporation process, which were related to changes in mass and viscous damping, respectively. It was found that droplet evaporation from a sticky superhydrophobic surface followed a constant contact radius (CCR) mode in the early stage of evaporation and a combination of CCR and constant contact angle modes without a Cassie-Wenzel transition in the final stage. Furthermore, AAO membranes with larger pore sizes exhibited longer evaporation times, which were attributed to evaporative cooling at the droplet interface.

  2. Droplet ejection and sliding on a flapping film

    Directory of Open Access Journals (Sweden)

    Xi Chen

    2017-03-01

    Full Text Available Water recovery and subsequent reuse are required for human consumption as well as industrial, and agriculture applications. Moist air streams, such as cooling tower plumes and fog, represent opportunities for water harvesting. In this work, we investigate a flapping mechanism to increase droplet shedding on thin, hydrophobic films for two vibrational cases (e.g., ± 9 mm and 11 Hz; ± 2 mm and 100 Hz. Two main mechanisms removed water droplets from the flapping film: vibrational-induced coalescence/sliding and droplet ejection from the surface. Vibrations mobilized droplets on the flapping film, increasing the probability of coalescence with neighboring droplets leading to faster droplet growth. Droplet departure sizes of 1–2 mm were observed for flapping films, compared to 3–4 mm on stationary films, which solely relied on gravity for droplet removal. Additionally, flapping films exhibited lower percentage area coverage by water after a few seconds. The second removal mechanism, droplet ejection was analyzed with respect to surface wave formation and inertia. Smaller droplets (e.g., 1-mm diameter were ejected at a higher frequency which is associated with a higher acceleration. Kinetic energy of the water was the largest contributor to energy required to flap the film, and low energy inputs (i.e., 3.3 W/m2 were possible. Additionally, self-flapping films could enable novel water collection and condensation with minimal energy input.

  3. Electrostatic field and charge distribution in small charged dielectric droplets

    Science.gov (United States)

    Storozhev, V. B.

    2004-08-01

    The charge distribution in small dielectric droplets is calculated on the basis of continuum medium approximation. There are considered charged liquid spherical droplets of methanol in the range of nanometer sizes. The problem is solved by the following way. We find the free energy of some ion in dielectric droplet, which is a function of distribution of other ions in the droplet. The probability of location of the ion in some element of volume in the droplet is a function of its free energy in this element of volume. The same approach can be applied to other ions in the droplet. The obtained charge distribution differs considerably from the surface distribution. The curve of the charge distribution in the droplet as a function of radius has maximum near the surface. Relative concentration of charges in the vicinity of the center of the droplet does not equal to zero, and it is the higher, the less is the total charge of the droplet. According to the estimates the model is applicable if the droplet radius is larger than 10 nm.

  4. Electrostatic field and charge distribution in small charged dielectric droplets

    International Nuclear Information System (INIS)

    Storozhev, V.B.

    2004-01-01

    The charge distribution in small dielectric droplets is calculated on the basis of continuum medium approximation. There are considered charged liquid spherical droplets of methanol in the range of nanometer sizes. The problem is solved by the following way. We find the free energy of some ion in dielectric droplet, which is a function of distribution of other ions in the droplet. The probability of location of the ion in some element of volume in the droplet is a function of its free energy in this element of volume. The same approach can be applied to other ions in the droplet. The obtained charge distribution differs considerably from the surface distribution. The curve of the charge distribution in the droplet as a function of radius has maximum near the surface. Relative concentration of charges in the vicinity of the center of the droplet does not equal to zero, and it is the higher, the less is the total charge of the droplet. According to the estimates the model is applicable if the droplet radius is larger than 10 nm

  5. Relationship between humidity and influenza A viability in droplets and implications for influenza's seasonality.

    Directory of Open Access Journals (Sweden)

    Wan Yang

    Full Text Available Humidity has been associated with influenza's seasonality, but the mechanisms underlying the relationship remain unclear. There is no consistent explanation for influenza's transmission patterns that applies to both temperate and tropical regions. This study aimed to determine the relationship between ambient humidity and viability of the influenza A virus (IAV during transmission between hosts and to explain the mechanisms underlying it. We measured the viability of IAV in droplets consisting of various model media, chosen to isolate effects of salts and proteins found in respiratory fluid, and in human mucus, at relative humidities (RH ranging from 17% to 100%. In all media and mucus, viability was highest when RH was either close to 100% or below ∼50%. When RH decreased from 84% to 50%, the relationship between viability and RH depended on droplet composition: viability decreased in saline solutions, did not change significantly in solutions supplemented with proteins, and increased dramatically in mucus. Additionally, viral decay increased linearly with salt concentration in saline solutions but not when they were supplemented with proteins. There appear to be three regimes of IAV viability in droplets, defined by humidity: physiological conditions (∼100% RH with high viability, concentrated conditions (50% to near 100% RH with lower viability depending on the composition of media, and dry conditions (<50% RH with high viability. This paradigm could help resolve conflicting findings in the literature on the relationship between IAV viability in aerosols and humidity, and results in human mucus could help explain influenza's seasonality in different regions.

  6. In vitro characterization of perfluorocarbon droplets for focused ultrasound therapy

    Energy Technology Data Exchange (ETDEWEB)

    Schad, Kelly C; Hynynen, Kullervo, E-mail: khynynen@sri.utoronto.c [Imaging Research, Sunnybrook Health Sciences Centre, 2075 Bayview Ave, Toronto, Ontario M4N 3M5 (Canada); Department of Medical Biophysics, University of Toronto (Canada)

    2010-09-07

    Focused ultrasound therapy can be enhanced with microbubbles by thermal and cavitation effects. However, localization of treatment is difficult as bioeffects can occur outside of the target region. Spatial control of bubbles can be achieved by ultrasound-induced conversion of liquid perfluorocarbon droplets to gas bubbles. This study was undertaken to determine the acoustic parameters for bubble production by droplet conversion and how it depends on the acoustic conditions and droplet physical parameters. Lipid-encapsulated droplets containing dodecafluoropentane were manufactured with sizes ranging from 1.9 to 7.2 {mu}m in diameter and diluted to a concentration of 8 x 10{sup 6} droplets mL{sup -1}. The droplets were sonicated in vitro with a focused ultrasound transducer and varying frequency and exposure under flow conditions through an acoustically transparent vessel. The sonications were 10 ms in duration at frequencies of 0.578, 1.736 and 2.855 MHz. The pressure threshold for droplet conversion was measured with an active transducer operating in pulse-echo mode and simultaneous measurements of broadband acoustic emissions were performed with passive acoustic detection. The results show that droplets cannot be converted at low frequency without broadband emissions occurring. However, the pressure threshold for droplet conversion decreased with increasing frequency, exposure and droplet size. The pressure threshold for broadband emissions was independent of the droplet size and was 2.9, 4.4 and 5.3 MPa for 0.578, 1736 and 2.855 MHz, respectively. In summary, we have demonstrated that droplet conversion is feasible for clinically relevant sized droplets and acoustic exposures.

  7. In vitro characterization of perfluorocarbon droplets for focused ultrasound therapy

    International Nuclear Information System (INIS)

    Schad, Kelly C; Hynynen, Kullervo

    2010-01-01

    Focused ultrasound therapy can be enhanced with microbubbles by thermal and cavitation effects. However, localization of treatment is difficult as bioeffects can occur outside of the target region. Spatial control of bubbles can be achieved by ultrasound-induced conversion of liquid perfluorocarbon droplets to gas bubbles. This study was undertaken to determine the acoustic parameters for bubble production by droplet conversion and how it depends on the acoustic conditions and droplet physical parameters. Lipid-encapsulated droplets containing dodecafluoropentane were manufactured with sizes ranging from 1.9 to 7.2 μm in diameter and diluted to a concentration of 8 x 10 6 droplets mL -1 . The droplets were sonicated in vitro with a focused ultrasound transducer and varying frequency and exposure under flow conditions through an acoustically transparent vessel. The sonications were 10 ms in duration at frequencies of 0.578, 1.736 and 2.855 MHz. The pressure threshold for droplet conversion was measured with an active transducer operating in pulse-echo mode and simultaneous measurements of broadband acoustic emissions were performed with passive acoustic detection. The results show that droplets cannot be converted at low frequency without broadband emissions occurring. However, the pressure threshold for droplet conversion decreased with increasing frequency, exposure and droplet size. The pressure threshold for broadband emissions was independent of the droplet size and was 2.9, 4.4 and 5.3 MPa for 0.578, 1736 and 2.855 MHz, respectively. In summary, we have demonstrated that droplet conversion is feasible for clinically relevant sized droplets and acoustic exposures.

  8. In vitro characterization of perfluorocarbon droplets for focused ultrasound therapy

    Science.gov (United States)

    Schad, Kelly C.; Hynynen, Kullervo

    2010-09-01

    Focused ultrasound therapy can be enhanced with microbubbles by thermal and cavitation effects. However, localization of treatment is difficult as bioeffects can occur outside of the target region. Spatial control of bubbles can be achieved by ultrasound-induced conversion of liquid perfluorocarbon droplets to gas bubbles. This study was undertaken to determine the acoustic parameters for bubble production by droplet conversion and how it depends on the acoustic conditions and droplet physical parameters. Lipid-encapsulated droplets containing dodecafluoropentane were manufactured with sizes ranging from 1.9 to 7.2 µm in diameter and diluted to a concentration of 8 × 106 droplets mL-1. The droplets were sonicated in vitro with a focused ultrasound transducer and varying frequency and exposure under flow conditions through an acoustically transparent vessel. The sonications were 10 ms in duration at frequencies of 0.578, 1.736 and 2.855 MHz. The pressure threshold for droplet conversion was measured with an active transducer operating in pulse-echo mode and simultaneous measurements of broadband acoustic emissions were performed with passive acoustic detection. The results show that droplets cannot be converted at low frequency without broadband emissions occurring. However, the pressure threshold for droplet conversion decreased with increasing frequency, exposure and droplet size. The pressure threshold for broadband emissions was independent of the droplet size and was 2.9, 4.4 and 5.3 MPa for 0.578, 1736 and 2.855 MHz, respectively. In summary, we have demonstrated that droplet conversion is feasible for clinically relevant sized droplets and acoustic exposures.

  9. The effect of bound to dialdehudecellulose protein concentration on the activity of immobilized trypsin after γ-irradiation and in process of storage

    International Nuclear Information System (INIS)

    Belov, A.A.; Ryl'tsev, V.V.; Ignatyuk, T.E.; Filatov, V.N.

    1994-01-01

    It is found the complex effect of the bound enzyme concentration on the proteolytic activity of trypsin immobilized to dialdehydecellulose (preriodate oxidation) after γ-irradiation and in process of storage. It is shown the occurance of three stages of immobilized enzyme inactivation in process of immobilization and storage. The velocity of inactivation did not depend on bound trypsin concentration. The ratio of proteolytic activity of samples before and after γ-irradiation was increased with the increase of immobilized to carrier enzyme concentration and was not change (in range of experiment error) in process of storage. The results were compared with that of cryctlline trypsin

  10. Probe Storage

    NARCIS (Netherlands)

    Gemelli, Marcellino; Abelmann, Leon; Engelen, Johannes Bernardus Charles; Khatib, M.G.; Koelmans, W.W.; Zaboronski, Olog; Campardo, Giovanni; Tiziani, Federico; Laculo, Massimo

    2011-01-01

    This chapter gives an overview of probe-based data storage research over the last three decades, encompassing all aspects of a probe recording system. Following the division found in all mechanically addressed storage systems, the different subsystems (media, read/write heads, positioning, data

  11. Proteomic Analysis of Lipid Droplets from Arabidopsis Aging Leaves Brings New Insight into Their Biogenesis and Functions

    Directory of Open Access Journals (Sweden)

    Lysiane Brocard

    2017-05-01

    Full Text Available Lipid droplets (LDs are cell compartments specialized for oil storage. Although their role and biogenesis are relatively well documented in seeds, little is known about their composition, structure and function in senescing leaves where they also accumulate. Here, we used a label free quantitative mass spectrometry approach to define the LD proteome of aging Arabidopsis leaves. We found that its composition is highly different from that of seed/cotyledon and identified 28 proteins including 9 enzymes of the secondary metabolism pathways involved in plant defense response. With the exception of the TRIGALACTOSYLDIACYLGLYCEROL2 protein, we did not identify enzymes implicated in lipid metabolism, suggesting that growth of leaf LDs does not occur by local lipid synthesis but rather through contact sites with the endoplasmic reticulum (ER or other membranes. The two most abundant proteins of the leaf LDs are the CALEOSIN3 and the SMALL RUBBER PARTICLE1 (AtSRP1; both proteins have structural functions and participate in plant response to stress. CALEOSIN3 and AtSRP1 are part of larger protein families, yet no other members were enriched in the LD proteome suggesting a specific role of both proteins in aging leaves. We thus examined the function of AtSRP1 at this developmental stage and found that AtSRP1 modulates the expression of CALEOSIN3 in aging leaves. Furthermore, AtSRP1 overexpression induces the accumulation of triacylglycerol with an unusual composition compared to wild-type. We demonstrate that, although AtSRP1 expression is naturally increased in wild type senescing leaves, its overexpression in senescent transgenic lines induces an over-accumulation of LDs organized in clusters at restricted sites of the ER. Conversely, atsrp1 knock-down mutants displayed fewer but larger LDs. Together our results reveal that the abundancy of AtSRP1 regulates the neo-formation of LDs during senescence. Using electron tomography, we further provide evidence

  12. ABHD5/CGI-58, the Chanarin-Dorfman Syndrome Protein, Mobilises Lipid Stores for Hepatitis C Virus Production.

    Directory of Open Access Journals (Sweden)

    Gabrielle Vieyres

    2016-04-01

    Full Text Available Hepatitis C virus (HCV particles closely mimic human very-low-density lipoproteins (VLDL to evade humoral immunity and to facilitate cell entry. However, the principles that govern HCV association with VLDL components are poorly defined. Using an siRNA screen, we identified ABHD5 (α/β hydrolase domain containing protein 5, also known as CGI-58 as a new host factor promoting both virus assembly and release. ABHD5 associated with lipid droplets and triggered their hydrolysis. Importantly, ABHD5 Chanarin-Dorfman syndrome mutants responsible for a rare lipid storage disorder in humans were mislocalised, and unable to consume lipid droplets or support HCV production. Additional ABHD5 mutagenesis revealed a novel tribasic motif that does not influence subcellular localization but determines both ABHD5 lipolytic and proviral properties. These results indicate that HCV taps into the lipid droplet triglyceride reservoir usurping ABHD5 lipase cofactor function. They also suggest that the resulting lipid flux, normally devoted to VLDL synthesis, also participates in the assembly and release of the HCV lipo-viro-particle. Altogether, our study provides the first association between the Chanarin-Dorfman syndrome protein and an infectious disease and sheds light on the hepatic manifestations of this rare genetic disorder as well as on HCV morphogenesis.

  13. Murine Diacylglycerol Acyltransferase-2 (DGAT2) Can Catalyze Triacylglycerol Synthesis and Promote Lipid Droplet Formation Independent of Its Localization to the Endoplasmic Reticulum*

    Science.gov (United States)

    McFie, Pamela J.; Banman, Shanna L.; Kary, Steven; Stone, Scot J.

    2011-01-01

    Triacylglycerol (TG) is the major form of stored energy in eukaryotic organisms and is synthesized by two distinct acyl-CoA:diacylglycerol acyltransferase (DGAT) enzymes, DGAT1 and DGAT2. Both DGAT enzymes reside in the endoplasmic reticulum (ER), but DGAT2 also co-localizes with mitochondria and lipid droplets. In this report, we demonstrate that murine DGAT2 is part of a multimeric complex consisting of several DGAT2 subunits. We also identified the region of DGAT2 responsible for its localization to the ER. A DGAT2 mutant lacking both its transmembrane domains, although still associated with membranes, was absent from the ER and instead localized to mitochondria. Unexpectedly, this mutant was still active and capable of interacting with lipid droplets to promote TG storage. Additional experiments indicated that the ER targeting signal was present in the first transmembrane domain (TMD1) of DGAT2. When fused to a fluorescent reporter, TMD1, but not TMD2, was sufficient to target mCherry to the ER. Finally, the interaction of DGAT2 with lipid droplets was dependent on the C terminus of DGAT2. DGAT2 mutants, in which regions of the C terminus were either truncated or specific regions were deleted, failed to co-localize with lipid droplets when cells were oleate loaded to stimulate TG synthesis. Our findings demonstrate that DGAT2 is capable of catalyzing TG synthesis and promote its storage in cytosolic lipid droplets independent of its localization in the ER. PMID:21680734

  14. Phase rainbow refractometry for accurate droplet variation characterization.

    Science.gov (United States)

    Wu, Yingchun; Promvongsa, Jantarat; Saengkaew, Sawitree; Wu, Xuecheng; Chen, Jia; Gréhan, Gérard

    2016-10-15

    We developed a one-dimensional phase rainbow refractometer for the accurate trans-dimensional measurements of droplet size on the micrometer scale as well as the tiny droplet diameter variations at the nanoscale. The dependence of the phase shift of the rainbow ripple structures on the droplet variations is revealed. The phase-shifting rainbow image is recorded by a telecentric one-dimensional rainbow imaging system. Experiments on the evaporating monodispersed droplet stream show that the phase rainbow refractometer can measure the tiny droplet diameter changes down to tens of nanometers. This one-dimensional phase rainbow refractometer is capable of measuring the droplet refractive index and diameter, as well as variations.

  15. Printing microstructures in a polymer matrix using a ferrofluid droplet

    International Nuclear Information System (INIS)

    Abdel Fattah, Abdel Rahman; Ghosh, Suvojit; Puri, Ishwar K.

    2016-01-01

    We print complex curvilinear microstructures in an elastomer matrix using a ferrofluid droplet as the print head. A magnetic field moves the droplet along a prescribed path in liquid polydimethylsiloxane (PDMS). The droplet sheds magnetic nanoparticle (MNP) clusters in its wake, forming printed features. The PDMS is subsequently heated so that it crosslinks, which preserves the printed features in the elastomer matrix. The competition between magnetic and drag forces experienced by the ferrofluid droplet and its trailing MNPs highlight design criteria for successful printing, which are experimentally confirmed. The method promises new applications, such as flexible 3D circuitry. - Highlights: • Magnetically guided miscible ferrofluid droplets print 3D patterns in a polymer. • Printing mechanism depends on the dynamics between the fluid and magnetic forces. • Droplet size influences the width of the printed trail. • The Colloidal distribution of the ferrofluid is important for pattern integrity. • Particle trajectories and trails are simulated and validated through experiments.

  16. Stochastic growth of cloud droplets by collisions during settling

    Science.gov (United States)

    Madival, Deepak G.

    2018-04-01

    In the last stage of droplet growth in clouds which leads to drizzle formation, larger droplets begin to settle under gravity and collide and coalesce with smaller droplets in their path. In this article, we shall deal with the simplified problem of a large drop settling amidst a population of identical smaller droplets. We present an expression for the probability that a given large drop suffers a given number of collisions, for a general statistically homogeneous distribution of droplets. We hope that our approach will serve as a valuable tool in dealing with droplet distribution in real clouds, which has been found to deviate from the idealized Poisson distribution due to mechanisms such as inertial clustering.

  17. Printing microstructures in a polymer matrix using a ferrofluid droplet

    Energy Technology Data Exchange (ETDEWEB)

    Abdel Fattah, Abdel Rahman [Department of Mechanical Engineering, Hamilton, Ontario (Canada); Ghosh, Suvojit [Department of Engineering Physics, McMaster University, Hamilton, Ontario (Canada); Puri, Ishwar K. [Department of Mechanical Engineering, Hamilton, Ontario (Canada); Department of Engineering Physics, McMaster University, Hamilton, Ontario (Canada)

    2016-03-01

    We print complex curvilinear microstructures in an elastomer matrix using a ferrofluid droplet as the print head. A magnetic field moves the droplet along a prescribed path in liquid polydimethylsiloxane (PDMS). The droplet sheds magnetic nanoparticle (MNP) clusters in its wake, forming printed features. The PDMS is subsequently heated so that it crosslinks, which preserves the printed features in the elastomer matrix. The competition between magnetic and drag forces experienced by the ferrofluid droplet and its trailing MNPs highlight design criteria for successful printing, which are experimentally confirmed. The method promises new applications, such as flexible 3D circuitry. - Highlights: • Magnetically guided miscible ferrofluid droplets print 3D patterns in a polymer. • Printing mechanism depends on the dynamics between the fluid and magnetic forces. • Droplet size influences the width of the printed trail. • The Colloidal distribution of the ferrofluid is important for pattern integrity. • Particle trajectories and trails are simulated and validated through experiments.

  18. The phospholipase PNPLA7 functions as a lysophosphatidylcholine hydrolase and interacts with lipid droplets through its catalytic domain.

    Science.gov (United States)

    Heier, Christoph; Kien, Benedikt; Huang, Feifei; Eichmann, Thomas O; Xie, Hao; Zechner, Rudolf; Chang, Ping-An

    2017-11-17

    Mammalian patatin-like phospholipase domain-containing proteins (PNPLAs) are lipid-metabolizing enzymes with essential roles in energy metabolism, skin barrier development, and brain function. A detailed annotation of enzymatic activities and structure-function relationships remains an important prerequisite to understand PNPLA functions in (patho-)physiology, for example, in disorders such as neutral lipid storage disease, non-alcoholic fatty liver disease, and neurodegenerative syndromes. In this study, we characterized the structural features controlling the subcellular localization and enzymatic activity of PNPLA7, a poorly annotated phospholipase linked to insulin signaling and energy metabolism. We show that PNPLA7 is an endoplasmic reticulum (ER) transmembrane protein that specifically promotes hydrolysis of lysophosphatidylcholine in mammalian cells. We found that transmembrane and regulatory domains in the PNPLA7 N-terminal region cooperate to regulate ER targeting but are dispensable for substrate hydrolysis. Enzymatic activity is instead mediated by the C-terminal domain, which maintains full catalytic competence even in the absence of N-terminal regions. Upon elevated fatty acid flux, the catalytic domain targets cellular lipid droplets and promotes interactions of PNPLA7 with these organelles in response to increased cAMP levels. We conclude that PNPLA7 acts as an ER-anchored lysophosphatidylcholine hydrolase that is composed of specific functional domains mediating catalytic activity, subcellular positioning, and interactions with cellular organelles. Our study provides critical structural insights into an evolutionarily conserved class of phospholipid-metabolizing enzymes. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  19. Trace of a water droplet exerted by coulomb force. 2

    International Nuclear Information System (INIS)

    Sugita, Hideaki; Murakami, Takuro; Nakazawa, Takeshi; Nakasako, Makoto; Yoshimura, Takuma; Osarakawa, Toshihiro

    2002-01-01

    The movement of water droplets in the air-water separator is based on the principle of the electrostatic precipitator with positive and negative poles. The mechanism of separation is that the water droplets charged negative ions or electrons by corona discharge are collected on the positive pole by Coulomb force operating between the both poles. This paper describes the theoretical analyses that how the movement of a water droplet is affected by Coulomb force in the air-water separator. (author)

  20. Droplet-Wall/Film Impact in IC Engine Applications

    Science.gov (United States)

    2017-08-14

    Report: Droplet-Wall/ Film Impact in IC Engine Applications (ARO Topic 1.4.1 under ARO’s Dr. Ralph A. Anthenien) The views, opinions and/or findings...Participants: RPPR Final Report as of 12-Oct-2017 Agreement Number: W911NF-16-1-0449 Organization: Princeton University Title: Droplet-Wall/ Film Impact...droplets impacting a wet surface under various film thickness, which plays a critical role in controlling the efficiency of applications such as those