Antigiardial activity of Ocimum basilicum essential oil.
de Almeida, Igor; Alviano, Daniela Sales; Vieira, Danielle Pereira; Alves, Péricles Barreto; Blank, Arie Fitzgerald; Lopes, Angela Hampshire C S; Alviano, Celuta Sales; Rosa, Maria do Socorro S
2007-07-01
In this study, we investigated the effects of Ocimum basilicum essential oil on Giardia lamblia and on the modulation of the interaction of these parasites by peritoneal mouse macrophage. The essential oil (2 mg/ml) and its purified substances demonstrated antigiardial activity. Linalool (300 microg/ml), however, was able to kill 100% parasites after 1 h of incubation, which demonstrates its high antigiardial potential. Pretreatment of peritoneal mouse macrophages with 2 mg/ml essential oil dilution reduced in 79% the association index between these macrophages and G. lamblia, with a concomitant increase by 153% on nitric oxide production by the G. lamblia-ingested macrophages. The protein profiles and proteolitic activity of these parasite trophozoites, previously treated or not with 2 mg/ml essential oil or with the purified fractions, were also determined. After 1 and 2 h of incubation, proteins of lysates and culture supernatants revealed significant differences in bands patterns when compared to controls. Besides, the proteolitic activity, mainly of cysteine proteases, was clearly inhibited by the essential oil (2 mg/ml) and the purified linalool (300 microg/ml). These results suggest that, with G. lamblia, the essential oil from O. basilicum and its purified compounds, specially linalool, have a potent antimicrobial activity.
Proteolytic activity of selected moulds in the first fermentation of black-seeded soysauce
NASA Astrophysics Data System (ADS)
Yulifianti, R.; Ginting, E.
2018-01-01
Black-seeded soybean is preferred as normally it has higher protein content and would give black colour to the soyfiltrate for making soysauce. Mould is usually used in the first fermentation of soysauce making to prepare koji with high soluble protein as a media for the subsequent bacteria fermentation in brine solution. Black-seeded soybean of Detam 1 variety was used as the soysauce ingredient. The trial was a randomized complete design with four replicates. The treatments were (1) Rhizopus oligosporus starter (in flour form) with two day-fermentation, (2) similar R. oligosporus starter with three-day fermentation, (3) Aspergillus sojae (pure culture) with three-day fermentation and (4) A. sojae in flour form with three-day fermentation. The black-seeded soybean had 100-grain weight of 11.7 g, high protein content of 42.5% dw and fat content of 14.9% dw. Koji prepared using R. oligosporus starter with two-day fermentation had the lowest protein content (48.9% dw). Both starters of A. sojae culture and flour gave the highest soluble protein content of koji (41.0-41.5% dw), followed by R. oligosporus starter with 3 day-fermentation (35.2% dw). Whilst the lowest value was noted in koji prepared using R. oligosporus starter with two day-fermentation (30.8% dw). This suggests that both A. sojae starters had similar proteolitic activity and higher than that of R. oligosporus starter. In terms of practical application and maintenance of the mould viability by soysauce processors, the use of A. sojae flour starter with three-day fermentation is suggested.
Palatsi, J; Viñas, M; Guivernau, M; Fernandez, B; Flotats, X
2011-02-01
Fresh pig/cattle slaughterhouse waste mixtures, with different lipid-protein ratios, were characterized and their anaerobic biodegradability assessed in batch tests. The resultant methane potentials were high (270-300 L(CH4) kg(-1)(COD)) making them interesting substrates for the anaerobic digestion process. However, when increasing substrate concentrations in consecutive batch tests, up to 15 g(COD) kg(-1), a clear inhibitory process was monitored. Despite the reported severe inhibition, related to lipid content, the system was able to recover activity and successfully degrade the substrate. Furthermore, 16SrRNA gene-based DGGE results showed an enrichment of specialized microbial populations, such as β-oxidizing/proteolitic bacteria (Syntrophomonas sp., Coprothermobacter sp. and Anaerobaculum sp.), and syntrophic methanogens (Methanosarcina sp.). Consequently, the lipid concentration of substrate and the structure of the microbial community are the main limiting factors for a successful anaerobic treatment of fresh slaughterhouse waste. Copyright © 2010 Elsevier Ltd. All rights reserved.
Carolina Rojas, O; León-Cachón, Rafael B R; Pérez-Maya, Antonio Alí; Aguirre-Garza, Marcelino; Moreno-Treviño, María G; González, Gloria M
2015-05-01
Chromoblastomycosis is a chronic granulomatous disease caused frequently by fungi of the Fonsecaea genus. The objective of this study was the phenotypic and molecular identification of F. pedrosoi strains isolated from chromoblastomycosis patients in Mexico and Venezuela. Ten strains were included in this study. For phenotypic identification, we used macroscopic and microscopic morphologies, carbohydrate assimilation test, urea hydrolysis, cixcloheximide tolerance, proteolitic activity and the thermotolerance test. The antifungal activity of five drugs was evaluated against the isolates. Molecular identification was performed by sequencing the internal transcribed spacer (ITS) ribosomal DNA regions of the isolated strains. The physiological analysis and morphological features were variable and the precise identification was not possible. All isolates were susceptible to itraconazole, terbinafine, voriconazole and posaconazole. Amphotericin B was the least effective drug. The alignment of the 559-nucleotide ITS sequences from our strains compared with sequences of GenBank revealed high homology with F. pedrosoi (EU285266.1). In this study, all patients were from rural areas, six from Mexico and four from Venezuela. Ten isolates were identified by phenotypic and molecular analysis, using ITS sequence and demonstrated that nine isolates from Mexico and Venezuela were 100% homologous and one isolate showed a small genetic distance. © 2015 Blackwell Verlag GmbH.
Ghahremani, Enayat; Mardani, Mahnaz; Rezapour, Sadegh
2015-03-01
Lactic acid bacteria (LAB) with proteolitic activity are used as aromatic and antibacterial substances, cholesterol reduces, bile salt hydrolyses, and probiotic. The aims of this project were to isolate and identify natural LAB flora involved in traditional fermentation in cheeses of Khoramabad city and also to survey their probiotic potential. In order to achieve this goal, LAB were isolated and characterized using phenotypic and genotypic methods (PCR-sequencing); in the next stage, they were analyzed lowering cholesterol medium, hydrolysis of the bile, resistance to bile-resistant PH acidic stomach. At the end of the study, 88 cocci and 3 bacill were found: 58 Enterococcus faecium, 16 Enterococcus hirae, 5 Lactococcus lactis, 3 Lactobacillus plantarum, and 9 undetermined. The probiotic results of the bacteria had effects on the reduction of cholesterol, resistance to stomach acid, had relative antibacterial effects, and some strains had effects on hydrolyzing the bile. For further identification, the PCR method and the application of 16s-DNA-ITS genes and its sequencing were found useful. This study showed that lactic acid bacteria in the traditional cheese of the Khorramabad city have relative probiotic effect and that these lactic acid bacteria in fermented milk are suitable.
Kurihara, Takamasa; Min, Jun Zhe; Hirata, Asuka; Toyo'oka, Toshimasa; Inagaki, Shinsuke
2009-05-01
Rapid, selective and sensitive determination of N-linked oligosaccharides in glycoproteins (ovalbumin, ribonuclease B and fetuin) was performed by ultra-performance liquid chromatography (UPLC) with fluorescence (FL) and electrospray ionization time-of-flight mass spectrometry (ESI-TOF-MS). The asparaginyl-oligosaccharide moiety was first liberated from each glycoprotein by pronase E (a proteolitic enzyme). The oligosaccharide fractions separated by gel-permeation chromatography were labeled with 1-pyrenesulfonyl chloride (PSC, a fluorescence reagent), separated by UPLC in a short run time, and then detected by FL and TOF-MS. The PSC-labeled oligosaccharides were selectively identified from the FL detection and then sensitively determined by ESI-TOF-MS. As the results, 15, eight and four kinds of N-linked oligosaccharides were detected from ovalbumin, ribonuclease B and fetuin, respectively. Because the present method is rapid (within 9 min), selective and sensitive (approximate 60 fmol, S/N = 5), the determination of N-linked oligosaccharides in various glycoproteins seems to be possible.
NASA Astrophysics Data System (ADS)
Ningrum, R. A.; Santoso, A.; Herawati, N.
2017-05-01
Human interferon alpha2a (hIFNα2a) is a therapeutic protein that used in cancer and hepatitis B/C therapy. The main problem of using hIFNα-2a is its short elimination half life due to its low molecular weight. Development of higher molecular weight protein by albumin fusion technology is a rational strategy to solve the problem. In our previous research we constructed an open reading frame (ORF) encoding hIFNα2a-human serum albumin (HSA) fusion protein that expressed in Pichia pastoris (P. pastoris) protease deficient strain SMD1168. This research was performed to overproduce, purify and characterize the fusion protein. To overproduce the protein, cultivation was performed in buffered complex medium containing glyserol (BMGY) for 24 h and protein overproduction was applied in buffered complex medium containing methanol (BMMY) for 48 hours at 30°C. The fusion protein was purified by blue sepharose affinity chromatography. Molecular weight characterization by SDS PAGE corresponds with its theoretical size, 85 kDa. Western blot analysis demonstrated that the fusion protein was recognized by anti hIFNα2 and anti HSA monoclonal antibody as well. Amino acid sequence of the fusion protein was determined by LC MS/MS2 mass spectrometry with trypsin as proteolitic enzyme. There were three fragments that identified as hIFNα2a and seven fragments that identified as HSA. Total identified amino acids were 150 residues with 20% coverage from total residues. To conclude, hIFNα2a-HSA fusion protein was overproduced, purified and characterized. Characterization based on molecular weight, antibody recognition and amino acid sequence confirmed that the fusion protein has correct identity as theoretically thought.
McCoy, G D
1980-03-01
The aniline hydroxylase activity of microsomes isolated from hamster liver can be differentiated kinetically into high affinity (low K(m), form I) and low affinity (high K(m), form II) forms. Microsomes isolated from uninduced animals contain slightly more form I activity. The activity of the low affinity form (form II) is preferentially enhanced by Aroclor or 3-methylcholanthrene treatment, while phenobarbital treatment increases the activity of both forms. Chronic ethanol consumption results in enhancement of only the high affinity form (form I).
Kanetsuna, Fuminori; Carbonell, Luis M.
1966-01-01
Kanetsuna, Fuminori (Instituto Venezolano de Investigaciones Cientificas, Caracas, Venezuela), and Luis M. Carbonell. Enzymes in glycolysis and the citric acid cycle in the yeast and mycelial forms of Paracoccidioides brasiliensis. J. Bacteriol. 92:1315–1320. 1966.—Enzymatic activities in glycolysis, the hexose monophosphate shunt, and the citric acid cycle in cell-free extracts of the yeast and mycelial forms of Paracoccidioides brasiliensis were examined comparatively. Both forms have the enzymes of these pathways. Activities of glucose-6-phosphate dehydrogenase and malic dehydrogenase of the mycelial form were higher than those of the yeast form. Another 15 enzymatic activities of the mycelial form were lower than those of the yeast form. The activity of glyceraldehyde-3-phosphate dehydrogenase showed the most marked difference between the two forms, its activity in the mycelial form being about 20% of that in the yeast form. PMID:5924267
Taboada-Serrano, Patricia; Tsouris, Constantino; Contescu, Cristian I; McFarlane, Joanna
2013-10-08
The present invention provides magnetically responsive activated carbon, and a method of forming magnetically responsive activated carbon. The method of forming magnetically responsive activated carbon typically includes providing activated carbon in a solution containing ions of ferrite forming elements, wherein at least one of the ferrite forming elements has an oxidation state of +3 and at least a second of the ferrite forming elements has an oxidation state of +2, and increasing pH of the solution to precipitate particles of ferrite that bond to the activated carbon, wherein the activated carbon having the ferrite particles bonded thereto have a positive magnetic susceptibility. The present invention also provides a method of filtering waste water using magnetic activated carbon.
The Built Environment and Active Travel: Evidence from Nanjing, China.
Feng, Jianxi
2016-03-08
An established relationship exists between the built environment and active travel. Nevertheless, the literature examining the impacts of different components of the built environment is limited. In addition, most existing studies are based on data from cities in the U.S. and Western Europe. The situation in Chinese cities remains largely unknown. Based on data from Nanjing, China, this study explicitly examines the influences of two components of the built environment--the neighborhood form and street form--on residents' active travel. Binary logistic regression analyses examined the effects of the neighborhood form and street form on subsistence, maintenance and discretionary travel, respectively. For each travel purpose, three models are explored: a model with only socio-demographics, a model with variables of the neighborhood form and a complete model with all variables. The model fit indicator, Nagelkerke's ρ², increased by 0.024 when neighborhood form variables are included and increased by 0.070 when street form variables are taken into account. A similar situation can be found in the models of maintenance activities and discretionary activities. Regarding specific variables, very limited significant impacts of the neighborhood form variables are observed, while almost all of the characteristics of the street form show significant influences on active transport. In Nanjing, street form factors have a more profound influence on active travel than neighborhood form factors. The focal point of the land use regulations and policy of local governments should shift from the neighborhood form to the street form to maximize the effects of policy interventions.
Tokunaga, Hiroko; Maeda, Junpei; Arakawa, Tsutomu; Tokunaga, Masao
2017-06-01
Effects of a water-miscible organic solvent, methanol, on the structure and activity of halophilic β-lactamase derived from Chromohalobacter sp.560 (HaBla), were investigated by means of circular dichroism (CD) measurement and enzymatic activity determination. Beta-lactamase activity was enhanced about 1.2-fold in the presence of 10-20% methanol. CD measurement of HaBla revealed different structures depending on the methanol concentration: native-like active form (Form I) in 10-20% methanol and methanol-induced inactive form at higher concentration (Form II in 40-60% and Form III in 75-80% methanol). Incubation of HaBla with 40% methanol led to the complete loss of activity within ~80 min accompanied by the formation of Form II, whose activity was recovered promptly up to ~80% of full activity upon dilution of the methanol concentration to 10%. In addition, when the protein concentration was sufficiently high (e.g., 0.7 mg/ml), HaBla activity of Form III in 75% methanol could be recovered in the same way (with slightly slower recovery rate), upon dilution of the methanol concentration. In contrast, non-halophilic β-lactamase from Escherichia coli K12 strain MG1655 (EcBla) was irreversibly denatured in the presence of 40% methanol. HaBla showed remarkable ability to renature from the methanol-induced inactive states.
The Built Environment and Active Travel: Evidence from Nanjing, China
Feng, Jianxi
2016-01-01
Background: An established relationship exists between the built environment and active travel. Nevertheless, the literature examining the impacts of different components of the built environment is limited. In addition, most existing studies are based on data from cities in the U.S. and Western Europe. The situation in Chinese cities remains largely unknown. Based on data from Nanjing, China, this study explicitly examines the influences of two components of the built environment—the neighborhood form and street form—on residents’ active travel. Methods: Binary logistic regression analyses examined the effects of the neighborhood form and street form on subsistence, maintenance and discretionary travel, respectively. For each travel purpose, three models are explored: a model with only socio-demographics, a model with variables of the neighborhood form and a complete model with all variables. Results: The model fit indicator, Nagelkerke’s ρ2, increased by 0.024 when neighborhood form variables are included and increased by 0.070 when street form variables are taken into account. A similar situation can be found in the models of maintenance activities and discretionary activities. Regarding specific variables, very limited significant impacts of the neighborhood form variables are observed, while almost all of the characteristics of the street form show significant influences on active transport. Conclusions: In Nanjing, street form factors have a more profound influence on active travel than neighborhood form factors. The focal point of the land use regulations and policy of local governments should shift from the neighborhood form to the street form to maximize the effects of policy interventions. PMID:27005645
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Lombraña, M; Suárez, P; San Juan, F
2005-09-01
alpha-Amylase activity has been shown for the first time in a non-digestive tissue from Mytilus galloprovincialis. alpha-amylase from mussel mantle tissue has been purified by affinity chromatography on insoluble starch, followed by gel-filtration chromatography on Superdex-200. The chromatographic and electrophoretic behaviour of M. galloprovincialis alpha-amylase and stability characteristics suggest two forms of this enzyme: one form forming stable aggregates (form I) and a monomeric form (form II) that is more abundant, active and unstable. Both forms show an inverse quantitative variation. Purified form II was highly unstable and the molecular mass was estimated to be 66 kDa by sodium dodecyl sulphate (SDS)-gel electrophoresis. Maximum activity was noted at pH 6.5 and 35 degrees C.
Loschiavo, F; Giarrizzo, S
1977-01-01
L Forms derived from strains of coagulase positive Staphylococcus aureus, have, on the whole, preserved their DNAsic, haemolitic and coagulastic activities. L. forms showed high resistence to antibiotics acting on the bacterial cell-wall. The sensibility to other antibiotics was, roughly, analogous for the L forms as well as for the bacterial strains ones, with the exception of the clortetraciclin and the diidrostreptomicin, ehich proved to be comparatively more active on the L forms.
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Intracellular proteolysis of pancreatic zymogens.
Gorelick, F. S.; Modlin, I. M.; Leach, S. D.; Carangelo, R.; Katz, M.
1992-01-01
Activation of pancreatic digestive zymogens within the pancreatic acinar cell may be an early event in the development of pancreatitis. To detect such activation, an immunoblot assay has been developed that measures the relative amounts of inactive zymogens and their respective active enzyme forms. Using this assay, high doses of cholecystokinin or carbachol were found to stimulate the intracellular conversion of at least three zymogens (procarboxypeptidase A1, procarboxypeptidase B, and chymotrypsinogen 2) to their active forms. Thus, this conversion may be a generalized phenomenon of pancreatic zymogens. The conversion is detected within ten minutes of treatment and is not associated with changes in acinar cell morphology; it has been predicted that the lysosomal thiol protease, cathepsin B, may initiate this conversion. Small amounts of cathepsin B are found in the secretory pathway, and cathepsin B can activate trypsinogen in vitro; however, exposure of acini to a thiol protease inhibitor (E64) did not block this conversion. Conversion was inhibited by the serine protease inhibitor, benzamidine, and by raising the intracellular pH, using chloroquine or monensin. This limited proteolytic conversion appears to require a low pH compartment and a serine protease activity. After long periods of treatment (60 minutes), the amounts of the active enzyme forms began to decrease; this observation suggested that the active enzyme forms were being degraded. Treatment of acini with E64 reduced this late decrease in active enzyme forms, suggesting that thiol proteases, including lysosomal hydrolases, may be involved in the degradation of the active enzyme forms. These findings indicate that pathways for zymogen activation as well as degradation of active enzyme forms are present within the pancreatic acinar cell. Images FIG. 1 FIG. 6 PMID:1340058
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Peeters, Elien; Hooyberghs, Geert; Robijns, Stijn; Waldrant, Kai; De Weerdt, Ami; Delattin, Nicolas; Liebens, Veerle; Kucharíková, Soňa; Tournu, Hélène; Verstraeten, Natalie; Dovgan, Barbara; Girandon, Lenart; Fröhlich, Mirjam; De Brucker, Katrijn; Michiels, Jan; Cammue, Bruno P. A.; Thevissen, Karin; Vanderleyden, Jozef; Van der Eycken, Erik
2016-01-01
We previously synthesized several series of compounds, based on the 5-aryl-2-aminoimidazole scaffold, that showed activity preventing the formation of Salmonella enterica serovar Typhimurium and Pseudomonas aeruginosa biofilms. Here, we further studied the activity spectrum of a number of the most active N1- and 2N-substituted 5-aryl-2-aminoimidazoles against a broad panel of biofilms formed by monospecies and mixed species of bacteria and fungi. An N1-substituted compound showed very strong activity against the biofilms formed by Gram-negative and Gram-positive bacteria and the fungus Candida albicans but was previously shown to be toxic against various eukaryotic cell lines. In contrast, 2N-substituted compounds were nontoxic and active against biofilms formed by Gram-negative bacteria and C. albicans but had reduced activity against biofilms formed by Gram-positive bacteria. In an attempt to develop nontoxic compounds with potent activity against biofilms formed by Gram-positive bacteria for application in antibiofilm coatings for medical implants, we synthesized novel compounds with substituents at both the N1 and 2N positions and tested these compounds for antibiofilm activity and toxicity. Interestingly, most of these N1-,2N-disubstituted 5-aryl-2-aminoimidazoles showed very strong activity against biofilms formed by Gram-positive bacteria and C. albicans in various setups with biofilms formed by monospecies and mixed species but lost activity against biofilms formed by Gram-negative bacteria. In light of application of these compounds as anti-infective coatings on orthopedic implants, toxicity against two bone cell lines and the functionality of these cells were tested. The N1-,2N-disubstituted 5-aryl-2-aminoimidazoles in general did not affect the viability of bone cells and even induced calcium deposition. This indicates that modulating the substitution pattern on positions N1 and 2N of the 5-aryl-2-aminoimidazole scaffold allows fine-tuning of both the antibiofilm activity spectrum and toxicity. PMID:27550355
Multi-layer coatings for bipolar rechargeable batteries with enhanced terminal voltage
Farmer, Joseph C.; Kaschmitter, James; Pierce, Steve
2017-06-06
A method for producing a multi-layer bipolar coated cell according to one embodiment includes applying a first active cathode material above a substrate to form a first cathode; applying a first solid-phase ionically-conductive electrolyte material above the first cathode to form a first electrode separation layer; applying a first active anode material above the first electrode separation layer to form a first anode; applying an electrically conductive barrier layer above the first anode; applying a second active cathode material above the anode material to form a second cathode; applying a second solid-phase ionically-conductive electrolyte material above the second cathode to form a second electrode separation layer; applying a second active anode material above the second electrode separation layer to form a second anode; and applying a metal material above the second anode to form a metal coating section. In another embodiment, the anode is formed prior to the cathode. Cells are also disclosed.
Back contact buffer layer for thin-film solar cells
Compaan, Alvin D.; Plotnikov, Victor V.
2014-09-09
A photovoltaic cell structure is disclosed that includes a buffer/passivation layer at a CdTe/Back contact interface. The buffer/passivation layer is formed from the same material that forms the n-type semiconductor active layer. In one embodiment, the buffer layer and the n-type semiconductor active layer are formed from cadmium sulfide (CdS). A method of forming a photovoltaic cell includes the step of forming the semiconductor active layers and the buffer/passivation layer within the same deposition chamber and using the same material source.
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Core-6 fucose and the oligomerization of the 1918 pandemic influenza viral neuraminidase
DOE Office of Scientific and Technical Information (OSTI.GOV)
Wu, Zhengliang L., E-mail: Leon.wu@bio-techne.com; Zhou, Hui; Ethen, Cheryl M.
The 1918 H1N1 influenza virus was responsible for one of the most deadly pandemics in human history. Yet to date, the structure component responsible for its virulence is still a mystery. In order to search for such a component, the neuraminidase (NA) antigen of the virus was expressed, which led to the discovery of an active form (tetramer) and an inactive form (dimer and monomer) of the protein due to different glycosylation. In this report, the N-glycans from both forms were released and characterized by mass spectrometry. It was found that the glycans from the active form had 26% core-6more » fucosylated, while the glycans from the inactive form had 82% core-6 fucosylated. Even more surprisingly, the stalk region of the active form was almost completely devoid of core-6-linked fucose. These findings were further supported by the results obtained from in vitro incorporation of azido fucose and {sup 3}H-labeled fucose using core-6 fucosyltransferase, FUT8. In addition, the incorporation of fucose did not change the enzymatic activity of the active form, implying that core-6 fucose is not directly involved in the enzymatic activity. It is postulated that core-6 fucose prohibits the oligomerization and subsequent activation of the enzyme. - Graphical abstract: Proposed mechanism for how core-fucose prohibits the tetramerization of the 1918 pandemic viral neuraminidase. Only the cross section of the stalk region with two N-linked glycans are depicted for clarity. (A) Carbohydrate–carbohydrate interaction on non-fucosylated monomer allows tetramerization. (B) Core-fucosylation disrupts the interaction and prevents the tetramerization. - Highlights: • Expressed 1918 pandemic influenza viral neuraminidase has inactive and active forms. • The inactive form contains high level of core-6 fucose, while the active form lacks such modification. • Core fucose could interfere the oligomerization of the neuraminidase and thus its activation. • This discovery may explain why 1918 pandemic influenza caused higher death rate among young population.« less
Yamada, Tsuyoshi; Kanda, Yutaka; Takayama, Makoto; Hashimoto, Akitoshi; Sugihara, Tsutomu; Satoh-Kubota, Ai; Suzuki-Takanami, Eri; Yano, Keiichi; Iida, Shigeru; Satoh, Mitsuo
2016-01-01
The structure of the N-linked oligosaccharides attached to antithrombin (AT) has been shown to affect its anticoagulant activity and pharmacokinetics. Human AT has biantennary complex-type oligosaccharides with the unique feature of lacking a core fucose, which affects its biological activities by changing its heparin-binding affinity. In human plasma, AT circulates as a mixture of the α-form bearing four oligosaccharides and the β-form lacking an oligosaccharide at Asn135. However, it remains unclear how the immature high-mannose-type oligosaccharides produced by mammalian cells affect biological activities of AT. Here, we succeeded in directly comparing the activities between the high-mannose and complex types. Interestingly, although there were no substantial differences in thrombin inhibitory activity, the high-mannose type showed higher heparin-binding affinity. The anticoagulant activities were increased by heparin and correlated with the heparin-binding affinity, resulting in the strongest anticoagulant activity being displayed in the β-form with the high-mannose type. In pharmacokinetic profiling, the high-mannose type showed a much shorter plasma half-life than the complex type. The β-form was found to have a prolonged plasma half-life compared with the α-form for the high-mannose type; conversely, the α-form showed a longer half-life than the β-form for the complex-type. The present study highlights that AT physiological activities are strictly controlled not only by a core fucose at the reducing end but also by the high-mannose-type structures at the nonreducing end. The β-form with the immature high-mannose type appears to function as a more potent anticoagulant than the AT typically found in human plasma, once it emerges in the blood. PMID:26747427
Yamada, Tsuyoshi; Kanda, Yutaka; Takayama, Makoto; Hashimoto, Akitoshi; Sugihara, Tsutomu; Satoh-Kubota, Ai; Suzuki-Takanami, Eri; Yano, Keiichi; Iida, Shigeru; Satoh, Mitsuo
2016-05-01
The structure of the N-linked oligosaccharides attached to antithrombin (AT) has been shown to affect its anticoagulant activity and pharmacokinetics. Human AT has biantennary complex-type oligosaccharides with the unique feature of lacking a core fucose, which affects its biological activities by changing its heparin-binding affinity. In human plasma, AT circulates as a mixture of the α-form bearing four oligosaccharides and the β-form lacking an oligosaccharide at Asn135. However, it remains unclear how the immature high-mannose-type oligosaccharides produced by mammalian cells affect biological activities of AT. Here, we succeeded in directly comparing the activities between the high-mannose and complex types. Interestingly, although there were no substantial differences in thrombin inhibitory activity, the high-mannose type showed higher heparin-binding affinity. The anticoagulant activities were increased by heparin and correlated with the heparin-binding affinity, resulting in the strongest anticoagulant activity being displayed in the β-form with the high-mannose type. In pharmacokinetic profiling, the high-mannose type showed a much shorter plasma half-life than the complex type. The β-form was found to have a prolonged plasma half-life compared with the α-form for the high-mannose type; conversely, the α-form showed a longer half-life than the β-form for the complex-type. The present study highlights that AT physiological activities are strictly controlled not only by a core fucose at the reducing end but also by the high-mannose-type structures at the nonreducing end. The β-form with the immature high-mannose type appears to function as a more potent anticoagulant than the AT typically found in human plasma, once it emerges in the blood. © The Author 2016. Published by Oxford University Press.
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The properties of B-form monoamine oxidase in mitochondria from monkey platelet.
Obata, Toshio; Aomine, Masahiro
The present study was examined the effect of the properties of monkey platelet monoamine oxidase (MAO) based on inhibitor sensitivity. Monkey platelet showed a high MAO activity with beta-phenylethylamine (beta-PEA) as substrate and a very low A-form MAO activity with 5 hydroxytryptamine (5-HT) as substrate. Moreover, monkey platelet MAO was sensitive to the drugs deprenyl as B-form MAO inhibitor and less sensitive to clorgyline and harmaline as A form MAO inhibitor with beta-PEA as the B-form MAO substrate. B-form MAO from monkey platelet was more stable against heat treatment at 55 degrees C than B-form MAO in brain. After digestion with trypsin at 37 degrees C for 4 hrs, it was found that MAO from platelet was inhibited about 70% with beta-PEA as substrate with brain. The tricyclic antidepressant imipramine and nortriptyline inhibited B-form MAO activity more potency than B-form MAO in brain. However, when the noncyclic antidepressant nomifensine was used, monkey platelet B-form MAO activities were less potently inhibited. All these reagents were noncompetitive inhibitors of B form MAO in monkey platelet. The present studies demonstrated that monkey platelet MAO is a single of B-form MAO and sensitive to tricyclic antidepressants.
USDA-ARS?s Scientific Manuscript database
The endogenous cellulase gene (CfEG3a) of Coptotermes formosanus, an economically important pest termite, was cloned and overexpressed in both native form (nCfEG) and C-terminal His-tagged form (tCfEG) in E.coli. Both forms of recombinant cellulases showed hydrolytic activity on cellulosic substrate...
76 FR 27079 - Agency Information Collection Activities: Declaration of Unaccompanied Articles
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Selective Suppression of Endothelial Cell Apoptosis by the High Molecular Weight Form of Adiponectin
Kobayashi, Hideki; Ouchi, Noriyuki; Kihara, Shinji; Walsh, Kenneth; Kumada, Masahiro; Abe, Yuki; Funahashi, Tohru; Matsuzawa, Yuji
2015-01-01
Adiponectin is an adipocyte-derived, antiatherogenic protein that is present in serum as three isoforms. Total adiponectin levels are decreased in obese or diabetic humans or animal models. This study was designed to elucidate the relative isoform distribution of adiponectin in human disease states and identify the active form of adiponectin toward vascular endothelial cells. The percentage of high molecular weight form (HMW) per total adiponectin was significantly lower in patients with coronary artery disease than control subjects, whereas the hexamer form was similar and the trimer form was significantly higher. During weight reduction in obese subjects, the HMW form increased and the trimer and hexamer forms decreased. Recombinant adiponectin dose-dependently suppressed apoptosis and caspase-3 activity in human umbilical vein endothelial cells (HUVECs). Transduction with dominant-negative AMP-activated protein kinase (AMPK) abolished the suppressive effect of adiponectin on HUVECs. Gel filtration chromatography was used to separate the adiponectin isoforms, and the antiapoptotic effect toward HUVECs was only observed with the HMW form. These data suggest that HMW adiponectin specifically confers the vascular-protective activities of this adipocytokine. PMID:14752031
Nanoporous carbon tunable resistor/transistor and methods of production thereof
Biener, Juergen; Baumann, Theodore F; Dasgupta, Subho; Hahn, Horst
2014-04-22
In one embodiment, a tunable resistor/transistor includes a porous material that is electrically coupled between a source electrode and a drain electrode, wherein the porous material acts as an active channel, an electrolyte solution saturating the active channel, the electrolyte solution being adapted for altering an electrical resistance of the active channel based on an applied electrochemical potential, wherein the active channel comprises nanoporous carbon arranged in a three-dimensional structure. In another embodiment, a method for forming the tunable resistor/transistor includes forming a source electrode, forming a drain electrode, and forming a monolithic nanoporous carbon material that acts as an active channel and selectively couples the source electrode to the drain electrode electrically. In any embodiment, the electrolyte solution saturating the nanoporous carbon active channel is adapted for altering an electrical resistance of the nanoporous carbon active channel based on an applied electrochemical potential.
Napper, A D; Bennett, S P; Borowski, M; Holdridge, M B; Leonard, M J; Rogers, E E; Duan, Y; Laursen, R A; Reinhold, B; Shames, S L
1994-01-01
A mixture of ananain (EC 3.4.22.31) and comosain purified from crude pineapple stem extract was found to contain numerous closely related enzyme forms. Chromatographic separation of the major enzyme forms was achieved after treatment of the mixture with thiol-modifying reagents: reversible modification with 2-hydroxyethyl disulphide provided enzyme for kinetic studies, and irreversible alkylation with bromotrifluoroacetone or iodoacetamide gave enzyme for structural analyses by 19F-n.m.r. and electrospray mass spectrometry respectively. Structural and kinetic analyses revealed comosain to be closely related to stem bromelain (EC 3.4.22.32), whereas ananain differed markedly from both comosain and stem bromelain. Nevertheless, differences were seen between comosain and stem bromelain in amino acid composition and kinetic specificity towards the epoxide inhibitor E-64. Differences between five isolatable alternative forms of ananain were characterized by amidolytic activity, thiol stoichiometry and accurate mass determinations. Three of the enzyme forms displayed ananain-like amidolytic activity, whereas the other two forms were inactive. Thiol-stoichiometry determinations revealed that the active enzyme forms contained one free thiol, whereas the inactive forms lacked the reactive thiol required for enzyme activity. M.s. provided direct evidence for oxidation of the active-site thiol to the corresponding sulphinic acid. Images Figure 3 Figure 4 PMID:8053898
Is there a core neural network in empathy? An fMRI based quantitative meta-analysis.
Fan, Yan; Duncan, Niall W; de Greck, Moritz; Northoff, Georg
2011-01-01
Whilst recent neuroimaging studies have identified a series of different brain regions as being involved in empathy, it remains unclear concerning the activation consistence of these brain regions and their specific functional roles. Using MKDA, a whole-brain based quantitative meta-analysis of recent fMRI studies of empathy was performed. This analysis identified the dACC-aMCC-SMA and bilateral anterior insula as being consistently activated in empathy. Hypothesizing that what are here termed affective-perceptual and cognitive-evaluative forms of empathy might be characterized by different activity patterns, the neural activations in these forms of empathy were compared. The dorsal aMCC was demonstrated to be recruited more frequently in the cognitive-evaluative form of empathy, whilst the right anterior insula was found to be involved in the affective-perceptual form of empathy only. The left anterior insula was active in both forms of empathy. It was concluded that the dACC-aMCC-SMA and bilateral insula can be considered as forming a core network in empathy, and that cognitive-evaluative and affective-perceptual empathy can be distinguished at the level of regional activation. Copyright © 2010 Elsevier Ltd. All rights reserved.
Durzyńska, Julia; Philippou, Anastassios; Brisson, Becky K.; Nguyen-McCarty, Michelle
2013-01-01
IGF-I is a key regulator of muscle development and growth. The pre-pro-peptide produced by the Igf1gene undergoes several posttranslational processing steps to result in a secreted mature protein, which is thought to be the obligate ligand for the IGF-I receptor (IGF-IR). The goals of this study were to determine what forms of IGF-I exist in skeletal muscle, and whether the mature IGF-I protein was the only form able to activate the IGF-IR. We measured the proportion of IGF-I species in murine skeletal muscle and found that the predominant forms were nonglycosylated pro-IGF-I and glycosylated pro-IGF-I, which retained the C-terminal E peptide extension, instead of mature IGF-I. These forms were validated using samples subjected to viral expression of IGF-I combined with furin and glycosidase digestion. To determine whether the larger molecular weight IGF-I forms were also ligands for the IGF-IR, we generated each specific form through transient transfection of 3T3 cells and used the enriched media to perform kinase receptor activation assays. Compared with mature IGF-I, nonglycosylated pro-IGF-I had similar ability to activate the IGF-IR, whereas glycosylation of pro-IGF-I significantly reduced receptor activation. Thus, it is important to understand not only the quantity, but also the proportion of IGF-I forms produced, to evaluate the true biological activity of this growth factor. PMID:23407451
Wegener, Gerhard; Macho, Claudia; Schlöder, Paul; Kamp, Günter; Ando, Osamu
2010-11-15
Trehalase (EC 3.2.1.28) hydrolyzes the main haemolymph sugar of insects, trehalose, into the essential cellular substrate glucose. Trehalase in locust flight muscle is bound to membranes that appear in the microsomal fraction upon tissue fractionation, but the exact location in vivo has remained elusive. Trehalase has been proposed to be regulated by a novel type of activity control that is based on the reversible transformation of a latent (inactive) form into an overt (active) form. Most trehalase activity from saline-injected controls was membrane-bound (95%) and comprised an overt form (∼25%) and a latent form (75%). Latent trehalase could be assayed only after the integrity of membranes had been destroyed. Trehazolin, a potent tight-binding inhibitor of trehalase, is confined to the extracellular space and has been used as a tool to gather information on the relationship between latent and overt trehalase. Trehazolin was injected into the haemolymph of locusts, and the trehalase activity of the flight muscle was determined at different times over a 30-day period. Total trehalase activity in locust flight muscle was markedly inhibited during the first half of the interval, but reappeared during the second half. Inhibition of the overt form preceded inhibition of the latent form, and the time course suggested a reversible precursor-product relation (cycling) between the two forms. The results support the working hypothesis that trehalase functions as an ectoenzyme, the activity of which is regulated by reversible transformation of latent into overt trehalase.
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2011-08-24
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Activated carbon fibers and engineered forms from renewable resources
Baker, Frederick S
2013-02-19
A method of producing activated carbon fibers (ACFs) includes the steps of providing a natural carbonaceous precursor fiber material, blending the carbonaceous precursor material with a chemical activation agent to form chemical agent-impregnated precursor fibers, spinning the chemical agent-impregnated precursor material into fibers, and thermally treating the chemical agent-impregnated precursor fibers. The carbonaceous precursor material is both carbonized and activated to form ACFs in a single step. The method produces ACFs exclusive of a step to isolate an intermediate carbon fiber.
Activated carbon fibers and engineered forms from renewable resources
Baker, Frederick S.
2010-06-01
A method of producing activated carbon fibers (ACFs) includes the steps of providing a natural carbonaceous precursor fiber material, blending the carbonaceous precursor material with a chemical activation agent to form chemical agent-impregnated precursor fibers, spinning the chemical agent-impregnated precursor material into fibers, and thermally treating the chemical agent-impregnated precursor fibers. The carbonaceous precursor material is both carbonized and activated to form ACFs in a single step. The method produces ACFs exclusive of a step to isolate an intermediate carbon fiber.
Code of Federal Regulations, 2010 CFR
2010-01-01
... Approval for Enterprise Products—Instructions and Notice of New Activity Form ER02JY09.000 ER02JY09.001... 12 Banks and Banking 7 2010-01-01 2010-01-01 false Prior Approval for Enterprise Products-Instructions and Notice of New Activity Form Appendix to Part 1253 Banks and Banking FEDERAL HOUSING FINANCE...
Brahimi-Adouane, Sabrina; Bachet, Jean-Baptiste; Tabone-Eglinger, Séverine; Subra, Frédéric; Capron, Claude; Blay, Jean-Yves; Emile, Jean-François
2013-06-01
Gain of function mutations of KIT are frequent in some human tumors, and are sensible to tyrosine kinase inhibitors. In most tumors, oncogenic mutations are heterozygous, however most in vitro data of KIT activation have been obtained with hemizygous mutation. This study aimed to investigate the maturation and activation of wild-type (WT) and mutant (M) forms of KIT in hemizygous and heterozygous conditions. WT and two types of exon 11 deletions M forms of human KIT were expressed in NIH3T3 cell lines. Membrane expression of KIT was quantified by flow cytometry. Quantification of glycosylated forms of KIT and phosphorylated forms of AKT and ERK were performed by western blot. Simultaneous activation of WT KIT and treatment with endoplasmic reticulum (ER) inhibitors, tunicamycin or brefeldin A induced a complete inhibition of membrane expression of the 145 kDa form of KIT. By contrast activation or ER inhibitors alone, only partly inhibited this form. ER inhibitors also inhibited KIT activation-dependent phosphorylation of AKT and ERK1/2. Brefeldin A induced a complete down regulation of the 145 kDa form in hemizygous M, and induced an intra-cellular accumulation of the 125 kDa form in WT but not in hemizygous M. Heterozygous cells had glycosylation and response to ER inhibitors patterns more similar to WT than to hemizygous M. Phosphorylated AKT was reduced in hemizygous cells in comparison to WT KIT cells and heterozygous cells, and in the presence of brefeldin A in all cell lines. Effects of ER inhibitors are significantly different in hemizygous and heterozygous mutants. Differences in intra-cellular trafficking of KIT forms result in differences in downstream signaling pathways, and activation of PI3K/AKT pathway appears to be tied to the presence of the mature 145 kDa form of KIT at the membrane surface. Copyright © 2012 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
Activity associated with coronal mass ejections at solar minimum - SMM observations from 1984-1986
NASA Technical Reports Server (NTRS)
St. Cyr, O. C.; Webb, D. F.
1991-01-01
Seventy-three coronal mass ejections (CMEs) observed by the coronagraph aboard SMM between 1984 and 1986 were examined in order to determine the distribution of various forms of solar activity that were spatially and temporally associated with mass ejections during solar minimum phase. For each coronal mass ejection a speed was measured, and the departure time of the transient from the lower corona estimated. Other forms of solar activity that appeared within 45 deg longitude and 30 deg latitude of the mass ejection and within +/-90 min of its extrapolated departure time were explored. The statistical results of the analysis of these 73 CMEs are presented, and it is found that slightly less than half of them were infrequently associated with other forms of solar activity. It is suggested that the distribution of the various forms of activity related to CMEs does not change at different phases of the solar cycle. For those CMEs with associations, it is found that eruptive prominences and soft X-rays were the most likely forms of activity to accompany the appearance of mass ejections.
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2012-12-14
...-0001] Agency Information Collection Activities: Petition for Alien Fiance(e), Form Number I-129F... information collection. (2) Title of the Form/Collection: Petition for Alien Fiance(e). (3) Agency form number... petition for an alien fiance(e), spouse, or his/her children. (5) An estimate of the total number of...
cAMP-CRP acts as a key regulator for the viable but non-culturable state in Escherichia coli.
Nosho, Kazuki; Fukushima, Hiroko; Asai, Takehiro; Nishio, Masahiro; Takamaru, Reiko; Kobayashi-Kirschvink, Koseki Joseph; Ogawa, Tetsuhiro; Hidaka, Makoto; Masaki, Haruhiko
2018-03-01
A variety of bacteria, including Escherichia coli, are known to enter the viable but non-culturable (VBNC) state under various stress conditions. During this state, cells lose colony-forming activities on conventional agar plates while retaining signs of viability. Diverse environmental stresses including starvation induce the VBNC state. However, little is known about the genetic mechanism inducing this state. Here, we aimed to reveal the genetic determinants of the VBNC state of E. coli. We hypothesized that the VBNC state is a process wherein specific gene products important for colony formation are depleted during the extended period of stress conditions. If so, higher expression of these genes would maintain colony-forming activities, thereby restraining cells from entering the VBNC state. From an E. coli plasmid-encoded ORF library, we identified genes that were responsible for maintaining high colony-forming activities after exposure to starvation condition. Among these, cpdA encoding cAMP phosphodiesterase exhibited higher performance in the maintenance of colony-forming activities. As cpdA overexpression decreases intracellular cAMP, cAMP or its complex with cAMP-receptor protein (CRP) may negatively regulate colony-forming activities under stress conditions. We confirmed this using deletion mutants lacking adenylate cyclase or CRP. These mutants fully maintained colony-forming activities even after a long period of starvation, while wild-type cells lost most of this activity. Thus, we concluded that the lack of cAMP-CRP effectively retains high colony-forming activities, indicating that cAMP-CRP acts as a positive regulator necessary for the induction of the VBNC state in E. coli.
76 FR 11254 - Agency Information Collection Activities: Declaration of Unaccompanied Articles
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2011-03-01
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Atterwill, C K; Reid, J; Athayde, C M
1985-05-01
The effect of thyroid status on the postnatal development of the two molecular forms of Na+,K+-ATPase, distinguished kinetically on the basis of their ouabain sensitivity, was examined in rat brain. Hypothyroidism induced by PTU from day 1 postnatally significantly reduced the Na+,K+-ATPase activity in cerebellum (22-30 days) but not forebrain, whereas hyperthyroidism (T4 treatment from day 1) had no effect. The hypothyroidism-induced reduction in cerebellum was reflected by a 20-45% reduction in the activity of the alpha(+) form of Na+,K+-ATPase (high ouabain affinity) against control brains compared to a 60-70% reduction in the activity of the alpha form (low ouabain affinity). These results show that neonatally induced hypothyroidism leads to a selectively greater impairment of the ontogenesis of the activity of cerebellar alpha form of Na+,K+-ATPase. This may possibly reflect a retarded development of a selective cerebellar cell population containing predominantly the alpha form of the enzyme.
Xu, Xiang; Zhao, Jingyue; Xu, Zhen; Peng, Baozhen; Huang, Qiuhua; Arnold, Eddy; Ding, Jianping
2004-08-06
Isocitrate dehydrogenases (IDHs) catalyze the oxidative decarboxylation of isocitrate to alpha-ketoglutarate, and regulation of the enzymatic activity of IDHs is crucial for their biological functions. Bacterial IDHs are reversibly regulated by phosphorylation of a strictly conserved serine residue at the active site. Eukaryotic NADP-dependent IDHs (NADP-IDHs) have been shown to have diverse important biological functions; however, their regulatory mechanism remains unclear. Structural studies of human cytosolic NADP-IDH (HcIDH) in complex with NADP and in complex with NADP, isocitrate, and Ca2+ reveal three biologically relevant conformational states of the enzyme that differ substantially in the structure of the active site and in the overall structure. A structural segment at the active site that forms a conserved alpha-helix in all known NADP-IDH structures assumes a loop conformation in the open, inactive form of HcIDH; a partially unraveled alpha-helix in the semi-open, intermediate form; and an alpha-helix in the closed, active form. The side chain of Asp279 of this segment occupies the isocitrate-binding site and forms hydrogen bonds with Ser94 (the equivalent of the phosphorylation site in bacterial IDHs) in the inactive form and chelates the metal ion in the active form. The structural data led us to propose a novel self-regulatory mechanism for HcIDH that mimics the phosphorylation mechanism used by the bacterial homologs, consistent with biochemical and biological data. This mechanism might be applicable to other eukaryotic NADP-IDHs. The results also provide insights into the recognition and specificity of substrate and cofactor by eukaryotic NADP-IDHs.
Smoking Discriminately Changes the Serum Active and Non-Active Forms of Vitamin B12.
Shekoohi, Niloofar; Javanbakht, Mohammad Hassan; Sohrabi, Marjan; Zarei, Mahnaz; Mohammadi, Hamed; Djalali, Mahmoud
2017-06-01
Smoking may modify the appetite, and consequently affect nutrient intake and serum micronutrients. The effect of smoking on vitamin B12 status has been considered in several studies. The research proposed that organic nitrites, nitro oxide, cyanides, and isocyanides of cigarette smoke interfere with vitamin B12 metabolism, and convert it to inactive forms. This research was carried out to determine the serum level of active and inactive forms of vitamin B12 in male smokers in comparison with male nonsmokers. This is a case-control study, in which the participants were 85 male smokers and 85 male nonsmokers. The serum levels of total and active form of vitamin B12 were measured. Dietary intake was recorded by a quantitative food frequency questionnaire and one-day 24-hour dietary recall method. Independent two sample T test was used to compare quantitative variables between the case and control groups. The serum level of total vitamin B12 was not significantly different between two groups, but serum level of active form of vitamin B12 in the smoking group was significantly lower than non-smoking group (P<0.001). This is one of the first studies that evaluated the serum level of active form of vitamin B12 in smokers in the Iranian community. The results of this study identified that serum level of total vitamin B12 might be not different between smoking and non-smoking people, but the function of this vitamin is disturbed in the body of smokers through the reduction of serum level of active form of vitamin B12.
Orzołek, Aleksandra; Wysocki, Paweł; Strzeżek, Jerzy; Kordan, Władysław
2013-03-01
The antioxidant system in semen is composed of enzymes, low-molecular weight antioxidants and seminal plasma proteins. Loss of enzymatic activity of superoxide dismutase (SOD) during semen preservation may cause insufficient antioxidant defense of boar spermatozoa. The aim of this study was to isolate and characterize SOD molecular forms from spermatozoa and to describe changes in SOD activity in boar sperm during preservation at 16°C. Sperm extracts were prepared from fresh or diluted semen and used for SOD purification or activity measurement. Ion-exchange chromatography and gel filtration was used to purify SOD molecular forms. BTS, Dilu Cell, M III and Vitasem were used as diluents for 5-day storage of semen at +16°C. The molecular form of SOD released from spermatozoa after cold shock and homogenization had a molecular weight of approximately 67kDa. The activity of the SOD form was the highest at pH 10 within the temperature range between 20 and 45°C. The enzymatic activity of form released after cold shock was inhibited by H2O2 and diethyldithiocarbamate (DDC; by 65 and 40%, respectively). The SOD form released by homogenization was inhibited by H2O2 and DDC (40%). The molecular form released after urea treatment was a 30kDa protein with maximum activity at 20°C and pH 10. Enzymatic activity of this form was inhibited by H2O2 by 35%, DDC by 80% and 2-mercaptoethanol by 15%. The antigenic determinants of SOD isolated from boar seminal plasma and spermatozoa were similar to each other. Susceptibility of spermatozoa to cold shock increased during storage, but the differences between extenders were statistically non-significant. Copyright © 2013 Society for Biology of Reproduction & the Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn. Published by Elsevier Urban & Partner Sp. z o.o. All rights reserved.
1988-04-01
limitations -- taxes V . How to fill out required forms -- AF form 1401 Petty Cash/Refund Voucher -- AF form 2539 NAP Disbursement Request 4N VI. General...AND ACTIVITY CODES ACTIVITY ACTIVITY CODES TITLE CODES TITLE " C2 Lanes Food and Beverage J3 Outside School Program C3 Lanes Pro Shop J4 Comb Fed... Food and Beverage 05 Course Maintenance L z Education E Outdoor Recreation LI Education Services Program El Marina (On-Base) Facility M = Chaplain
Sebastião, E; Gobbi, S; Chodzko-Zajko, W; Schwingel, A; Papini, C B; Nakamura, P M; Netto, A V; Kokubun, E
2012-11-01
To explore issues associated with measuring physical activity using the International Physical Activity Questionnaire (IPAQ)-long form in adults living in a mid-sized Brazilian city. A stratified random sampling procedure was used to select a representative sample of adults living in Rio Claro. This yielded 1572 participants who were interviewed using the IPAQ-long form. The data were analysed using standard statistical procedures. Overall, 83% of men and 89% of women reported at least 150 min of combined moderate and/or vigorous physical activity per week. Reliable values of leisure and transportation-related physical activity were observed for both males and females. With regard to the household and work-related physical activity domains, both males and females reported unusually high levels of participation. The IPAQ-long form appears to overestimate levels of physical activity for both males and females, suggesting that the instrument has problems in measuring levels of physical activity in Brazilian adults. Accordingly, caution is warranted before using IPAQ data to support public policy decisions related to physical activity. Copyright © 2012 The Royal Society for Public Health. Published by Elsevier Ltd. All rights reserved.
76 FR 71057 - Agency Information Collection Activities: Crew's Effects Declaration
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2011-11-16
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78 FR 28820 - Commission Information Collection Activities (FERC Form 80); Comment Request; Revision
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2013-05-16
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2013-04-25
... Information Collection Activities (FERC Form 80); Comment Request; Extension AGENCY: Federal Energy Regulatory... automatic notification of activity in this docket or in viewing/downloading comments and issuances in this... jurisdiction to recognize the resources that are affected by their activities and to play a role in protecting...
Nora, Anni; Renvall, Hanna; Kim, Jeong-Young; Service, Elisabet; Salmelin, Riitta
2015-01-01
Temporal and frontal activations have been implicated in learning of novel word forms, but their specific roles remain poorly understood. The present magnetoencephalography (MEG) study examines the roles of these areas in processing newly-established word form representations. The cortical effects related to acquiring new phonological word forms during incidental learning were localized. Participants listened to and repeated back new word form stimuli that adhered to native phonology (Finnish pseudowords) or were foreign (Korean words), with a subset of the stimuli recurring four times. Subsequently, a modified 1-back task and a recognition task addressed whether the activations modulated by learning were related to planning for overt articulation, while parametrically added noise probed reliance on developing memory representations during effortful perception. Learning resulted in decreased left superior temporal and increased bilateral frontal premotor activation for familiar compared to new items. The left temporal learning effect persisted in all tasks and was strongest when stimuli were embedded in intermediate noise. In the noisy conditions, native phonotactics evoked overall enhanced left temporal activation. In contrast, the frontal learning effects were present only in conditions requiring overt repetition and were more pronounced for the foreign language. The results indicate a functional dissociation between temporal and frontal activations in learning new phonological word forms: the left superior temporal responses reflect activation of newly-established word-form representations, also during degraded sensory input, whereas the frontal premotor effects are related to planning for articulation and are not preserved in noise. PMID:25961571
Nora, Anni; Renvall, Hanna; Kim, Jeong-Young; Service, Elisabet; Salmelin, Riitta
2015-01-01
Temporal and frontal activations have been implicated in learning of novel word forms, but their specific roles remain poorly understood. The present magnetoencephalography (MEG) study examines the roles of these areas in processing newly-established word form representations. The cortical effects related to acquiring new phonological word forms during incidental learning were localized. Participants listened to and repeated back new word form stimuli that adhered to native phonology (Finnish pseudowords) or were foreign (Korean words), with a subset of the stimuli recurring four times. Subsequently, a modified 1-back task and a recognition task addressed whether the activations modulated by learning were related to planning for overt articulation, while parametrically added noise probed reliance on developing memory representations during effortful perception. Learning resulted in decreased left superior temporal and increased bilateral frontal premotor activation for familiar compared to new items. The left temporal learning effect persisted in all tasks and was strongest when stimuli were embedded in intermediate noise. In the noisy conditions, native phonotactics evoked overall enhanced left temporal activation. In contrast, the frontal learning effects were present only in conditions requiring overt repetition and were more pronounced for the foreign language. The results indicate a functional dissociation between temporal and frontal activations in learning new phonological word forms: the left superior temporal responses reflect activation of newly-established word-form representations, also during degraded sensory input, whereas the frontal premotor effects are related to planning for articulation and are not preserved in noise.
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2012-02-22
... Activities: Comment Request for the Nonferrous Metals Surveys (30 Forms) AGENCY: U.S. Geological Survey (USGS... revision of the currently approved paperwork requirements for the Nonferrous Metals Surveys. This... Control Number: 1028-0053. Form Number: Various (30 forms). Title: Nonferrous Metals Surveys. Type of...
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... Activities; Proposed Collection; Comment Request; Company-Run Annual Stress Test Reporting Template and... Federal savings associations titled, ``Company-Run Annual Stress Test Reporting Template and Documentation... Web site under Tools and Forms ( http://www.occ.gov/tools-forms/forms/bank-operations/stress-test...
78 FR 56241 - Agency Information Collection Activities:
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2013-09-12
...-0095] Agency Information Collection Activities: Notice of Appeal or Motion, Form I-290B; Revision of a... Collection (1) Type of Information Collection Request: Revision of a Currently Approved Collection. (2) Title of the Form/Collection: Notice of Appeal or Motion. (3) Agency form number, if any, and the...
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2011-06-08
... Advance Notification Form: Tourist and Other Non-Governmental Activities in the Antarctic Treaty Area... of Information Collection: Advance Notification Form: Tourist and Other Non-Governmental Activities... proposed information collection is necessary for the proper performance of our functions. Evaluate the...
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2010-02-01
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2011-06-20
... Collection (Disability Benefits Questionnaires--Group 3) Activity Under OMB Review AGENCY: Veterans Benefits... Questionnaire, VA Form 21-0960C-5. b. Headaches (Including Migraine Headaches), Disability Benefits Questionnaire, VA Form 21-0960C-8. c. Multiple Sclerosis (MS), Disability Benefits Questionnaire, VA Form 21...
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77 FR 6137 - Agency Information Collection Activities: Guam-CNMI Visa Waiver Agreement
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2012-02-07
... Activities: Guam-CNMI Visa Waiver Agreement AGENCY: U.S. Customs and Border Protection, Department of... comment on an information collection requirement concerning the Guam-CNMI Visa Waiver Agreement (CBP Form... information collection: Title: Guam-CNMI Visa Waiver Agreement. OMB Number: 1651-0126. Form Number: CBP Form I...
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2011-01-24
... automated collection techniques or other forms of information technology. A Comment to OMB is most effective.... NHTSA-2010-0023] Reports, Forms and Recordkeeping Requirements; Agency Information Collection Activity... announces that the Information Collection Request (ICR) abstracted below has been forwarded to the Office of...
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2013-12-09
... Activities: Canadian Border Boat Landing Permit (CBP Form I-68) AGENCY: U.S. Customs and Border Protection... information collection requirement concerning the Canadian Border Boat Landing Permit (Form I-68). This... soliciting comments concerning the following information collection: Title: Canadian Border Boat Landing...
77 FR 22068 - Proposed Information Collection (Financial Status Report) Activity: Comment Request
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2012-04-12
... (Financial Status Report) Activity: Comment Request AGENCY: Office of Management, Department of Veterans... techniques or the use of other forms of information technology. Title: Financial Status Report, VA Form 5655...: Claimants complete VA Form 5655 to report their financial status. VA uses the data collected to determine...
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2013-10-22
... DEPARTMENT OF HOMELAND SECURITY U.S. Citizenship and Immigration Services [OMB Control Number 1615-0043] Agency Information Collection Activities: Application for Temporary Protected Status, Form I-821... Currently Approved Collection. (2) Title of the Form/Collection: Application for Temporary Protected Status...
Erdner, Anette; Magnusson, Annabella
2012-10-01
It is well known that people with severe mental illness often suffer from constant fatigue, insomnia, and somatic complaints that are too often overlooked. In addition, these persons die earlier in life than others in the population. The purpose of this study was to investigate patients' descriptions of activities and the importance of these activities for their health. Eight persons living in their own home were interviewed about both their views about exercise and their exercising activities. Two themes emerged: Getting Control over One's Life and The Need for Contact with Family & Friends. All of the informants were aware of the importance of physical activity to feel good. The informants described three different forms of activities: daily activities in the home, activities in a rehabilitation centre, and various forms of jogging. These different forms of activity were important to the informants since they reduced their anxiety and stress.
Satagopan, Sriram; Chan, Sum; Perry, L. Jeanne; Tabita, F. Robert
2014-01-01
The first x-ray crystal structure has been solved for an activated transition-state analog-bound form II ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). This enzyme, from Rhodopseudomonas palustris, assembles as a unique hexamer with three pairs of catalytic large subunit homodimers around a central 3-fold symmetry axis. This oligomer arrangement is unique among all known Rubisco structures, including the form II homolog from Rhodospirillum rubrum. The presence of a transition-state analog in the active site locked the activated enzyme in a “closed” conformation and revealed the positions of critical active site residues during catalysis. Functional roles of two form II-specific residues (Ile165 and Met331) near the active site were examined via site-directed mutagenesis. Substitutions at these residues affect function but not the ability of the enzyme to assemble. Random mutagenesis and suppressor selection in a Rubisco deletion strain of Rhodobacter capsulatus identified a residue in the amino terminus of one subunit (Ala47) that compensated for a negative change near the active site of a neighboring subunit. In addition, substitution of the native carboxyl-terminal sequence with the last few dissimilar residues from the related R. rubrum homolog increased the enzyme's kcat for carboxylation. However, replacement of a longer carboxyl-terminal sequence with termini from either a form III or a form I enzyme, which varied both in length and sequence, resulted in complete loss of function. From these studies, it is evident that a number of subtle interactions near the active site and the carboxyl terminus account for functional differences between the different forms of Rubiscos found in nature. PMID:24942737
Satagopan, Sriram; Chan, Sum; Perry, L Jeanne; Tabita, F Robert
2014-08-01
The first x-ray crystal structure has been solved for an activated transition-state analog-bound form II ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). This enzyme, from Rhodopseudomonas palustris, assembles as a unique hexamer with three pairs of catalytic large subunit homodimers around a central 3-fold symmetry axis. This oligomer arrangement is unique among all known Rubisco structures, including the form II homolog from Rhodospirillum rubrum. The presence of a transition-state analog in the active site locked the activated enzyme in a "closed" conformation and revealed the positions of critical active site residues during catalysis. Functional roles of two form II-specific residues (Ile(165) and Met(331)) near the active site were examined via site-directed mutagenesis. Substitutions at these residues affect function but not the ability of the enzyme to assemble. Random mutagenesis and suppressor selection in a Rubisco deletion strain of Rhodobacter capsulatus identified a residue in the amino terminus of one subunit (Ala(47)) that compensated for a negative change near the active site of a neighboring subunit. In addition, substitution of the native carboxyl-terminal sequence with the last few dissimilar residues from the related R. rubrum homolog increased the enzyme's kcat for carboxylation. However, replacement of a longer carboxyl-terminal sequence with termini from either a form III or a form I enzyme, which varied both in length and sequence, resulted in complete loss of function. From these studies, it is evident that a number of subtle interactions near the active site and the carboxyl terminus account for functional differences between the different forms of Rubiscos found in nature. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.
This data collection form is divided into two parts: Child Activity Diary and Food Survey. The Child Activity Diary collects information on the child's activities at home over the 48-hr monitoring period. The diary is divided into four time periods over the 48-hr monitoring inter...
This data collection form is divided into two parts: Child Activity Diary and Food Survey. The Child Activity Diary collects information on the child's activities at home over the 48-hr monitoring period. The diary is divided into five time periods over the 48-hr monitoring inter...
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2013-07-30
... Information Collection Activities (FERC Form 80); Comment Request; Extension AGENCY: Federal Energy Regulatory... Office of Information and Regulatory Affairs: [email protected] . Attention: Federal Energy... receiving automatic notification of activity in this docket or in viewing/downloading comments and issuances...
Effect of chemical form of selenium on tissue glutathione peroxidase activity in developing rats
NASA Technical Reports Server (NTRS)
Lane, Helen W.; Strength, Ralph; Johnson, Janet; White, Marguerite T.
1991-01-01
The hypothesis that the stage of development of rats may affect the availability of various forms of selenium for the activity of glutathione peroxidase (GSHPx) in the rat was experimentally investigated. One experiment evaluated the availability of selenium as selenite or selenomethionine for GSPHx activity during three developmental states in rats: fetus and 7-day old and 14-day old nursing pups. In all tissues studied, GSHPx activity was highest in the 14-day-old pups whose dams were in the selenomethionine group. Rat pups given intraperitoneal selenite had higher liver and kidney GSHPx activity than pups given the same amount of selenium as intraperitoneal selenomethionine. In a second experiment, all dams were fed the same basal diet and pups were weaned to diets containing one of two levels of selenium and one of three forms of selenium (selenite, selenomethionine, or selenocystine). The results also supported the hypothesis these dietary forms of selenium are differentially available for GSHPx activity.
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2013-03-15
... Activities: Declaration for Free Entry of Unaccompanied Articles AGENCY: U.S. Customs and Border Protection...: Declaration for Free Entry of Unaccompanied Articles (Form 3299). This is a proposed extension of an... Unaccompanied Articles. OMB Number: 1651-0014. Form Number: Form 3299. Abstract: 19 U.S.C. 1498 provides that...
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2013-01-25
...-0113] Agency Information Collection Activities: InfoPass System, No Form Number; Extension, Without... Change, of a Currently Approved Collection. (2) Title of the Form/Collection: InfoPass System. (3) Agency...: Primary: Individuals or households. The InfoPass system allows an applicant or petitioner to schedule an...
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2013-01-25
...-0046] Agency Information Collection Activities: Inter-Agency Alien Witness and Informant Record, Form I... collection. (2) Title of the Form/Collection: Inter-Agency Alien Witness and Informant Record. (3) Agency...- 854 is used by law enforcement agencies to bring alien witnesses and informants to the United States...
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2011-08-09
... Activities: Form G-884, Extension of an Existing Information Collection; Comment Request ACTION: 30-Day Notice of Information Collection Under Review: Form G- 884, Request for the Return of Original Document(s... information technology, e.g., permitting electronic submission of responses. Overview of This Information...
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2012-02-28
... Collection Activities: Form G-28, Revision of a Currently Approved Information Collection; Comment Request ACTION: 30-Day Notice of Information Collection Under Review: Form G- 28, Notice of Entry of Appearance... technology, e.g., permitting electronic submission of responses. Overview of This information collection: (1...
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2011-05-11
... Collection Activity (Veteran Suicide Prevention Online Quantitative Surveys) Under OMB Review AGENCY... techniques or the use of other forms of information technology. Titles a. Veterans Online Survey, VA Form 10-0513. b. Veterans Family Online Survey, VA Form 10-0513a. c. Veterans Primary Care Provider Online...
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2011-08-22
... Activities: Petition for Remission or Mitigation of Forfeitures and Penalties Incurred AGENCY: U.S. Customs...: Petition for Remission or Mitigation of Forfeitures and Penalties Incurred (Form 4609). This is a proposed... Mitigation of Forfeitures and Penalties Incurred. OMB Number: 1651-0100. Form Number: CBP Form 4609. Abstract...
77 FR 19304 - Agency Information Collection Activities: Guam-CNMI Visa Waiver Information
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2012-03-30
... Activities: Guam-CNMI Visa Waiver Information AGENCY: U.S. Customs and Border Protection (CBP), Department of... requirement concerning the Guam-CNMI Visa Waiver Information (CBP Form I-736). This request for comment is...: Title: Guam-CNMI Visa Waiver Information. OMB Number: 1651-0109. Form Number: CBP Form I-736. Abstract...
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... Activities: Visa Waiver Program Carrier Agreement (CBP Form I-775) AGENCY: U.S. Customs and Border Protection... information collection requirement concerning the Visa Waiver Program Carrier Agreement (CBP Form I-775). This... concerning the following information collection: Title: Visa Waiver Program Carrier Agreement. OMB Number...
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2010-02-01
... Activities: Visa Waiver Program Carrier Agreement (Form I-775) AGENCY: U.S. Customs and Border Protection... and Budget (OMB) for review and approval in accordance with the Paperwork Reduction Act: Visa Waiver... other technological techniques or other forms of information. Title: Visa Waiver Program Carrier...
78 FR 6849 - Agency Information Collection (Verification of VA Benefits) Activity Under OMB Review
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2013-01-31
... (Verification of VA Benefits) Activity Under OMB Review AGENCY: Veterans Benefits Administration, Department of... ``OMB Control No. 2900-0406.'' SUPPLEMENTARY INFORMATION: Title: Verification of VA Benefits, VA Form 26... eliminate unlimited versions of lender- designed forms. The form also informs the lender whether or not the...
Blintsov, A N; Gussakovskaya, M A
2004-10-01
An original modification of the standard ELISA procedure for differential determination of different forms of abscisic acid (ABA) is proposed. It is shown that endogenous forms of ABA may be quantitatively determined in plant tissues subjected to minimal treatment, without purification of the hormones and their chemical modification. The modification has been approved when analyzing changes in the content of different ABA forms in plant tissues differing in physiological activity. Quantitative differential determination of changes in the content of different ABA forms has been performed in ovaries of Triticum aestivum L. and Taraxacum officinale Web. in the period of activity of the ovule (from the moment of its activation to the beginning of division). It is shown that, despite the different types of reproduction in the species studied (amphimixis and apomixis), the time course of changes in the content of different forms of ABA in ovaries is similar, which is suggestive of a correlation between the activity of endogenous hormonal system and chronology of main events (e.g., the beginning of endospermogenesis) of the reproductive cycle.
Physical Conditioning through Water Exercises.
ERIC Educational Resources Information Center
Conrad, C. Carson
This document describes activities in an aquatic program designed for an individual in sound health. Instructions for performing each activity are given in step-by-step outline form. The activities are arranged under the following categories: standing water drills; pool-side standing drills; gutter holding drills; bobbing (various forms);…
76 FR 34293 - Proposed Collection; Comment Request for Form 8582
Federal Register 2010, 2011, 2012, 2013, 2014
2011-06-13
... 8582, Passive Activity Loss Limitations. DATES: Written comments should be received on or before August... INFORMATION: Title: Passive Activity Loss Limitations. OMB Number: 1545-1008. Form Number: 8582. Abstract: Under Internal Revenue Code section 469, losses from passive activities, to the extent that they exceed...
Membrane Treatment of Aqueous Film Forming Foam (AFFF) Wastes for Recovery of Its Active Ingredients
1980-10-01
T ME1MBRANE TREATMENT OF AQUEOUS FILM FORMING FOAM~ (AFFF) WASTES FOR RECOVERY OFI Fts ACTIVE INGREDIENTS FINAL REPORT October 1980 by Edward S. K...OF THIS PAGEOPMn Date AVntr* d)__ ---- Ultrafiltration (UF) and Reverse Osmosis (RO) treatment of Aqueous Film Forming Foam (AFFF) solutions was...of Aqueous Film Forming Foam (AFFF) solutions was investigated to determine the feasibility of employing membrane processes to separate and recover
Nicolaus Taurellus on forms and elements.
Blank, Andreas
2014-12-01
This article examines the conception of elements in the natural philosophy of Nicolaus Taurellus (1547-1606) and explores the theological motivation that stands behind this conception. By some of his early modern readers, Taurellus may have been understood as a proponent of material atoms. By contrast, I argue that considerations concerning the substantiality of the ultimate constituents of composites led Taurellus to an immaterialist ontology, according to which elements are immaterial forms that possess active and passive potencies as well as motion and extension. In Taurellus's view, immaterialism about elements provides support for the theological doctrine of creation ex nihilo. As he argues, the ontology of immaterial forms helps to explicate a sense in which creatures are substances, not accidents of the divine substance. In particular, he maintains that immaterial forms stand in suitable relations of ontological dependence to God: creation dependence (since forms would not exist without the divine act of creation), but neither subsistence dependence (since forms continue to exist without continued divine agency) nor activity dependence (since forms are active without requiring divine concurrence).
ERIC Educational Resources Information Center
Marsden, Emma; Chen, Hsin-Ying
2011-01-01
This study aimed to isolate the effects of the two input activities in Processing Instruction: referential activities, which force learners to focus on a form and its meaning, and affective activities, which contain exemplars of the target form and require learners to process sentence meaning. One hundred and twenty 12-year-old Taiwanese learners…
Hall, Edward Thomas; Gray, Shirley; Sproule, John
2016-01-01
The activities and behaviours of a female head coach of a national rugby union team were recorded in both training and competition, across a whole rugby season, using the newly developed Rugby Coach Activities and Behaviours Instrument (RCABI). The instrument incorporates 24 categories of behaviour, embedded within three forms of activity (training form (TF), playing form (PF) and competitive match) and seven sub-activity types. In contrast to traditional drill-based coaching, 58.5% of the training time was found to have been spent in PF activities. Moreover, the proportion of PF activities increased to a peak average of 83.8% in proximity to the team's annual international championship. Uniquely, one of the coach's most prolific behaviours was conferring with associates (23.3%), highlighting the importance of interactions with assistant coaches, medical staff and others in shaping the coaching process. Additionally, the frequencies of key behaviours such as questioning and praise were found to vary between the different activity forms and types, raising questions about previous conceptions of effective coaching practice. The findings are discussed in the light of the Game Sense philosophy and the role of the head coach.
Underground waste barrier structure
Saha, Anuj J.; Grant, David C.
1988-01-01
Disclosed is an underground waste barrier structure that consists of waste material, a first container formed of activated carbonaceous material enclosing the waste material, a second container formed of zeolite enclosing the first container, and clay covering the second container. The underground waste barrier structure is constructed by forming a recessed area within the earth, lining the recessed area with a layer of clay, lining the clay with a layer of zeolite, lining the zeolite with a layer of activated carbonaceous material, placing the waste material within the lined recessed area, forming a ceiling over the waste material of a layer of activated carbonaceous material, a layer of zeolite, and a layer of clay, the layers in the ceiling cojoining with the respective layers forming the walls of the structure, and finally, covering the ceiling with earth.
Form-Focused Discovery Activities in English Classes
ERIC Educational Resources Information Center
Ogeyik, Muhlise Cosgun
2011-01-01
Form-focused discovery activities allow language learners to grasp various aspects of a target language by contributing implicit knowledge by using discovered explicit knowledge. Moreover, such activities can assist learners to perceive and discover the features of their language input. In foreign language teaching environments, they can be used…
Russian: An Active Introduction.
ERIC Educational Resources Information Center
De La Cruz, Nina
The Active Introduction is one of the modules in an array of materials used in Russian training for beginners at the Foreign Service Institute. It is essentially a catalog of sentences relating to typical daily activities which can be combined to form different communication sequences in dialog form. Students learn to speak Russian through…
Structure of the active form of human origin recognition complex and its ATPase motor module
DOE Office of Scientific and Technical Information (OSTI.GOV)
Tocilj, Ante; On, Kin Fan; Yuan, Zuanning
Binding of the Origin Recognition Complex (ORC) to origins of replication marks the first step in the initiation of replication of the genome in all eukaryotic cells. Here, we report the structure of the active form of human ORC determined by X-ray crystallography and cryo-electron microscopy. The complex is composed of an ORC1/4/5 motor module lobe in an organization reminiscent of the DNA polymerase clamp loader complexes. A second lobe contains the ORC2/3 subunits. The complex is organized as a double-layered shallow corkscrew, with the AAA+ and AAA+-like domains forming one layer, and the winged-helix domains (WHDs) forming a topmore » layer. CDC6 fits easily between ORC1 and ORC2, completing the ring and the DNA-binding channel, forming an additional ATP hydrolysis site. Analysis of the ATPase activity of the complex provides a basis for understanding ORC activity as well as molecular defects observed in Meier-Gorlin Syndrome mutations.« less
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O'Connor, Donna; Larkin, Paul; Williams, A Mark
2018-01-01
We used systematic observation tools to explore the structure (i.e., activity and inactivity) and sequencing (i.e., the types of activities used) of football coaching sessions in Australia following the implementation of a new National Curriculum. Youth soccer coaches (n = 34), coaching within the Skill Acquisition (U11-U13 n = 19) and Game Training (U14-U17 n = 15) phases of the Football Federation Australia National Curriculum participated. Participants were filmed during a regular coaching session, with systematic observation of the session undertaken to provide a detailed analysis of the practice activities and coach behaviours. Findings indicated a session comprised of Playing Form activities (40.9%), Training Form activities (22.3%), inactivity (31%), and transitions between activities (5.8%). Coaches prescribed more Training Form activities (e.g., individual (5.4%) and drills (15.1%)) early in the session and progressed to Playing Form activities (i.e., small-sided games (15.3%) then larger games (24.8%)) later in the session. Most inactivity reflected the players listening to the coach - either in a team huddle (9.9%) or frozen on the spot during an activity (16.5%). In addition, coaches generally spent over 3 min communicating to players prior to explaining and introducing an activity regardless of when in the session the activity was scheduled.
Normal form from biological motion despite impaired ventral stream function.
Gilaie-Dotan, S; Bentin, S; Harel, M; Rees, G; Saygin, A P
2011-04-01
We explored the extent to which biological motion perception depends on ventral stream integration by studying LG, an unusual case of developmental visual agnosia. LG has significant ventral stream processing deficits but no discernable structural cortical abnormality. LG's intermediate visual areas and object-sensitive regions exhibit abnormal activation during visual object perception, in contrast to area V5/MT+ which responds normally to visual motion (Gilaie-Dotan, Perry, Bonneh, Malach, & Bentin, 2009). Here, in three studies we used point light displays, which require visual integration, in adaptive threshold experiments to examine LG's ability to detect form from biological and non-biological motion cues. LG's ability to detect and discriminate form from biological motion was similar to healthy controls. In contrast, he was significantly deficient in processing form from non-biological motion. Thus, LG can rely on biological motion cues to perceive human forms, but is considerably impaired in extracting form from non-biological motion. Finally, we found that while LG viewed biological motion, activity in a network of brain regions associated with processing biological motion was functionally correlated with his V5/MT+ activity, indicating that normal inputs from V5/MT+ might suffice to activate his action perception system. These results indicate that processing of biologically moving form can dissociate from other form processing in the ventral pathway. Furthermore, the present results indicate that integrative ventral stream processing is necessary for uncompromised processing of non-biological form from motion. Copyright © 2011 Elsevier Ltd. All rights reserved.
New Forms of Student Activism: Lobbying, Trusteeing, and Collective Bargaining.
ERIC Educational Resources Information Center
Beeler, Kent D.
1979-01-01
This article focuses on three new forms of student activism: lobbying, trusteeing, and collective bargaining. Related aspects of student involvement in the political, legal, and consumer areas are discussed briefly. (Author)
Dolfi, Sonia C.; Yang, Zhihong; Lee, Mao-Jung; Guan, Fei; Hong, Jungil; Yang, Chung S.
2013-01-01
Tocopherols are the major source of dietary vitamin E. In this study, the growth inhibitory effects of different forms of tocopherols, tocopheryl phosphates (TP) and tocopherol quinones (TQ) on human colon cancer HCT116 and HT29 cells were investigated. δ-T was more active than γ-T in inhibiting colon cancer cell growth, decreasing cancer cell colony formation and inducing apoptosis; however α-T was rather ineffective. Similarly, the rate of cellular uptake also followed the ranking order δ-T > γ-T ≫ α-T. TP and TQ generally had higher inhibitory activities than their parent compounds. Interestingly, the γ-forms of TP and TQ were more active than the δ-forms in inhibiting cancer cell growth; whereas the α-forms were the least effective. The potencies of γ-TQ and δ-TQ (showing IC50 of ~0.8 and ~2 μM on HCT116 cells after a 72-h incubation, respectively) were >100 and >20 fold higher, respectively, than those of their parent tocopherols. Induction of cancer cell apoptosis by δ-T, γ-TP and γ-TQ was characterized by the cleavage of caspase 3 and PARP1 and DNA fragmentation. These studies demonstrated the higher growth inhibitory activity of δ-T than γ-T, the even higher activities of the γ-forms of TP and TQ, and the ineffectiveness of the α-forms of tocopherol and their metabolites against colon cancer cells. PMID:23898832
Dolfi, Sonia C; Yang, Zhihong; Lee, Mao-Jung; Guan, Fei; Hong, Jungil; Yang, Chung S
2013-09-11
Tocopherols are the major source of dietary vitamin E. In this study, the growth inhibitory effects of different forms of tocopherols (T), tocopheryl phosphates (TP), and tocopherol quinones (TQ) on human colon cancer HCT116 and HT29 cells were investigated. δ-T was more active than γ-T in inhibiting colon cancer cell growth, decreasing cancer cell colony formation, and inducing apoptosis; however, α-T was rather ineffective. Similarly, the rate of cellular uptake also followed the ranking order δ-T > γ-T ≫ α-T. TP and TQ generally had higher inhibitory activities than their parent compounds. Interestingly, the γ forms of TP and TQ were more active than the δ forms in inhibiting cancer cell growth, whereas the α forms were the least effective. The potencies of γ-TQ and δ-TQ (showing IC50 values of ∼0.8 and ∼2 μM on HCT116 cells after a 72 h incubation, respectively) were greater than 100-fold and greater than 20-fold higher, respectively, than those of their parent tocopherols. Induction of cancer cell apoptosis by δ-T, γ-TP, and γ-TQ was characterized by the cleavage of caspase 3 and PARP1 and DNA fragmentation. These studies demonstrated the higher growth inhibitory activity of δ-T than γ-T, the even higher activities of the γ forms of TP and TQ, and the ineffectiveness of the α forms of tocopherol and their metabolites against colon cancer cells.
ERIC Educational Resources Information Center
Redchenko, Nadezhda N.
2016-01-01
The authors of this article suggest a thesis about the purpose of teaching a foreign language--it is student's communicative activities, i.e. learning a foreign language in practice. The teacher's task is to encourage activities of every student and to create situations to develop their creative activities in a learning process. New information…
Sun, S; Karsdal, M A; Bay-Jensen, A C; Sørensen, M G; Zheng, Q; Dziegiel, M H; Maksymowych, W P; Henriksen, K
2013-10-01
Cathepsin K plays essential roles in bone resorption and is intensely investigated as a therapeutic target for the treatment of osteoporosis. Hence an assessment of the active form of cathepsin K may provide important biological information in metabolic bone diseases, such as osteoporosis or ankylosing spondylitis. Presently there are no robust assays for the assessment of active cathepsin K in serum, and therefore an ELISA specifically detecting the N-terminal of the active form of cathepsin K was developed. The assay was technically robust, with a lowest limit of detection (LOD) of 0.085 ng/mL. The average intra- and inter-assay CV% were 6.60% and 8.56% respectively. The dilution recovery and spike recovery tests in human serum were within 100±20% within the range of the assay. A comparison of latent and active cathepsin K confirmed specificity towards the active form. Quantification of the levels of active cathepsin K in supernatants of purified human osteoclasts compared to corresponding macrophages showed a 30-fold induction (p<0.001). In contrast, in serum samples from osteoporotic women on estrogen or bisphosphonate therapy and from ankylosing spondylitis patients no clinically relevant differences were observed. In summary, we have developed a robust and sensitive assay specifically detecting the active form of cathepsin K; however, while it monitors osteoclasts with high specificity in vitro, it appears that circulating levels of active cathepsin K do not reflect bone changes under these circumstances. Copyright © 2013 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.
Rulli, Karen; Yugawa, Takashi; Hanson, Charlotte; Thompson, Delores; Ruscetti, Sandra; Nishigaki, Kazuo
2004-01-01
The erythroleukemia-inducing Friend spleen focus-forming virus (SFFV) encodes a unique envelope protein, gp55, which interacts with the erythropoietin (Epo) receptor complex, causing proliferation and differentiation of erythroid cells in the absence of Epo. Susceptibility to SFFV-induced erythroleukemia is conferred by the Fv-2 gene, which encodes a short form of the receptor tyrosine kinase Stk/Ron (sf-Stk) only in susceptible strains of mice. We recently demonstrated that sf-Stk becomes activated by forming a strong interaction with SFFV gp55. To examine the biological consequences of activated sf-Stk on erythroid cell growth, we prepared retroviral vectors which express sf-Stk, either in conjunction with gp55 or alone in a constitutively activated mutant form, and tested them for their ability to induce Epo-independent erythroid colonies ex vivo and disease in mice. Our data indicate that both gp55-activated sf-Stk and the constitutively activated mutant of sf-Stk induce erythroid cells from Fv-2-susceptible and Fv-2-resistant (sf-Stk null) mice to form Epo-independent colonies. Mutational analysis of sf-Stk indicated that a functional kinase domain and 8 of its 12 tyrosine residues are required for the induction of Epo-independent colonies. Further studies demonstrated that coexpression of SFFV gp55 with sf-Stk significantly extends the half-life of the kinase. When injected into Fv-2-resistant mice, neither the gp55-activated sf-Stk nor the constitutively activated mutant caused erythroleukemia. Surprisingly, both Fv-2-susceptible and -resistant mice injected with the gp55-sf-Stk vector developed clinical signs not previously associated with SFFV-induced disease. We conclude that sf-Stk, activated by either point mutation or interaction with SFFV gp55, is sufficient to induce Epo-independent erythroid colonies from both Fv-2-susceptible and -resistant mice but is unable to cause erythroleukemia in Fv-2-resistant mice. PMID:15078939
Large-scale patterns formed by solar active regions during the ascending phase of cycle 21
NASA Astrophysics Data System (ADS)
Gaizauskas, V.; Harvey, K. L.; Harvey, J. W.; Zwaan, C.
1983-02-01
Synoptic maps of photospheric magnetic fields prepared at the Kitt Peak National Observatory are used in investigating large-scale patterns in the spatial and temporal distribution of solar active regions for 27 solar rotations between 1977 and 1979. The active regions are found to be distributed in 'complexes of activity' (Bumba and Howard, 1965). With the working definition of a complex of activity based on continuity and proximity of the constituent active regions, the phenomenology of complexes is explored. It is found that complexes of activity form within one month and that they are typically maintained for 3 to 6 solar rotations by fresh injections of magnetic flux. During the active lifetime of a complex of activity, the total magnetic flux in the complex remains steady to within a factor of 2. The magnetic polarities are closely balanced, and each complex rotates about the sun at its own special, constant rate. In certain cases, the complexes form two diverging branches.
Education Technologies in Addressing the Problem of Forming the Socially Active Individual
ERIC Educational Resources Information Center
Popova, Irina N.
2016-01-01
The article is devoted to the analysis of technological support of the educational process in solving the problem of forming the socially active individual. The authors studied the value of the category "social activity" and analyzed educational technologies that have an impact on its formation. The obtained results gave the possibility…
Utsumi, J; Yamazaki, S; Kawaguchi, K; Kimura, S; Shimizu, H
1989-10-05
Human interferon-beta 1 is extremely stable is a low ionic strength solution of pH 2 such as 10 mM HCl at 37 degrees C. However, the presence of 0.15 M NaCl led to a remarkable loss of antiviral activity. The molecular-sieve high-performance liquid chromatography revealed that, whereas completely active human interferon-beta 1 eluted as a 25 kDa species (monomeric form), the inactivated preparation eluted primarily as a 90 kDa species (oligomeric form). The specific activity (units per mg protein) of the oligomeric form was approx. 10% of that of the monomeric form. This observation shows that oligomeric human interferon-beta 1 is apparently in an inactive form. When the oligomeric eluate was resolved by polyacrylamide gel containing sodium dodecyl sulphate (SDS), it appeared to be monomeric under non-reducing conditions. Monomerization of the oligomeric human interferon-beta 1 by treatment with 1% SDS, fully regenerated its antiviral activity. These results suggest that the inactivation of the human interferon-beta 1 preparation was caused by its oligomerization via hydrophobic interactions without the formation of intermolecular disulphide bonds. These oligomers can be dissociated by SDS to restore biological activity.
A mechanism for the activation of the Na/H exchanger NHE-1 by cytoplasmic acidification and mitogens
Lacroix, Jérôme; Poët, Mallorie; Maehrel, Céline; Counillon, Laurent
2004-01-01
Eukaryotic cells constantly have to fight against internal acidification. In mammals, this task is mainly performed by the ubiquitously expressed electroneutral Na+/H+ exchanger NHE-1, which activates in a cooperative manner when cells become acidic. Despite its biological importance, the mechanism of this activation is still poorly understood, the most commonly accepted hypothesis being the existence of a proton-sensor site on the internal face of the transporter. This work uncovers mutations that lead to a nonallosteric form of the exchanger and demonstrates that NHE-1 activation is best described by a Monod–Wyman–Changeux concerted mechanism for a dimeric transporter. During intracellular acidification, a low-affinity form of NHE-1 is converted into a form possessing a higher affinity for intracellular protons, with no requirement for an additional proton-sensor site on the protein. This new mechanism also explains the activation of the exchanger by growth signals, which shift the equilibrium towards the high-affinity form. PMID:14710192
Freeman, Spencer A; McLeod, Sarah J; Dukowski, Janet; Austin, Pamela; Lee, Crystal C Y; Millen-Martin, Brandie; Kubes, Paul; McCafferty, Donna-Marie; Gold, Michael R; Roskelley, Calvin D
2010-06-01
The Rap1 GTPase is a master regulator of cell adhesion, polarity, and migration. We show that both blocking Rap1 activation and expressing a constitutively active form of Rap1 reduced the ability of B16F1 melanoma cells to extravasate from the microvasculature and form metastatic lesions in the lungs. This correlated with a decreased ability of the tumor cells to undergo transendothelial migration (TEM) in vitro and form dynamic, F-actin-rich pseudopodia that penetrate capillary endothelial walls in vivo. Using multiple tumor cell lines, we show that the inability to form these membrane protrusions, which likely promote TEM and extravasation, can be explained by altered adhesion dynamics and impaired cell polarization that result when Rap1 activation or cycling is perturbed. Thus, targeting Rap1 could be a useful approach for reducing the metastatic dissemination of tumor cells that undergo active TEM. Copyright 2010 AACR.
Simulation-Based Validation of the p53 Transcriptional Activity with Hybrid Functional Petri Net.
Doi, Atsushi; Nagasaki, Masao; Matsuno, Hiroshi; Miyano, Satoru
2011-01-01
MDM2 and p19ARF are essential proteins in cancer pathways forming a complex with protein p53 to control the transcriptional activity of protein p53. It is confirmed that protein p53 loses its transcriptional activity by forming the functional dimer with protein MDM2. However, it is still unclear that protein p53 keeps its transcriptional activity when it forms the trimer with proteins MDM2 and p19ARF. We have observed mutual behaviors among genes p53, MDM2, p19ARF and their products on a computational model with hybrid functional Petri net (HFPN) which is constructed based on information described in the literature. The simulation results suggested that protein p53 should have the transcriptional activity in the forms of the trimer of proteins p53, MDM2, and p19ARF. This paper also discusses the advantages of HFPN based modeling method in terms of pathway description for simulations.
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...] Agency Information Collection Activities: Proposed Collection; Comments Requested; Strategic Planning... Form/Collection: Strategic Planning Environmental Assessment Outreach. (3) Agency form number, if any... Strategic Planning Office at ATF will [[Page 32138
Potentiometric/turbidometric titration of antiperspirant actives.
Johnston, Clifford T; Hem, Stanley L; Guenin, Eric; Mattai, Jairajh; Afflito, John
2003-01-01
A titration procedure that simultaneously monitors the pH and turbidity of an antiperspirant solution during neutralization with sodium hydroxide was developed to characterize antiperspirant actives. Aluminum chloride, aluminum chlorohydrate (ACH), and aluminum zirconium glycine complex (AZG) gave distinctive pH/turbidity profiles. The activated forms of aluminum chlorohydrate (ACH') and aluminum zirconium glycine complex (AZG') produced more turbidity than the non-activated forms. On an equimolar basis, AZG' produced more turbidity than any of the antiperspirant actives tested.
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... ID No. BOEM-2011-0010] BOEMRE Information Collection Activity; 1010-0141, Subpart D, Oil and Gas... to oil and gas drilling operations, and related forms. DATES: Submit written comments by June 14..., Subpart D, Oil and Gas Drilling Operations. BOEMRE Form(s): MMS-123, MMS-123S, MMS-124, MMS-125, MMS-133...
Pore-forming activity of clostridial binary toxins.
Knapp, O; Benz, R; Popoff, M R
2016-03-01
Clostridial binary toxins (Clostridium perfringens Iota toxin, Clostridium difficile transferase, Clostridium spiroforme toxin, Clostridium botulinum C2 toxin) as Bacillus binary toxins, including Bacillus anthracis toxins consist of two independent proteins, one being the binding component which mediates the internalization into cell of the intracellularly active component. Clostridial binary toxins induce actin cytoskeleton disorganization through mono-ADP-ribosylation of globular actin and are responsible for enteric diseases. Clostridial and Bacillus binary toxins share structurally and functionally related binding components which recognize specific cell receptors, oligomerize, form pores in endocytic vesicle membrane, and mediate the transport of the enzymatic component into the cytosol. Binding components retain the global structure of pore-forming toxins (PFTs) from the cholesterol-dependent cytotoxin family such as perfringolysin. However, their pore-forming activity notably that of clostridial binding components is more related to that of heptameric PFT family including aerolysin and C. perfringens epsilon toxin. This review focuses upon pore-forming activity of clostridial binary toxins compared to other related PFTs. This article is part of a Special Issue entitled: Pore-Forming Toxins edited by Mauro Dalla Serra and Franco Gambale. Copyright © 2015 Elsevier B.V. All rights reserved.
Ohta, Kazumasa; Takahashi, Chifumi; Tosuji, Hiroaki
2009-08-01
The activity of acetylcholinesterase (AchE) increases rapidly after the gastrula stage of sea urchin development. In this report, changes in activity and in the molecular differentiation of AchE were investigated. AchE activity increased slightly during gastrulation and rose sharply thereafter, and was dependent on new RNA synthesis. No activity of butyrylcholinesterase was found. Morphogenesis in sea urchin embryos was inhibited by the AchE inhibitor eserine, which specifically inhibited arm rod formation but not body rod formation. Spicule formation and enzyme activity in cultured micromeres were inhibited by eserine in a dose-dependent manner. During gastrulation, two molecular forms of AchE were detected with polyacrylamide gel electrophoresis. The appearance of an additional band on the gel was consistent with the occurrence of a remarkable increase in the enzyme activity. This additional band appeared as a larger molecular form in Anthocidaris crassispina, Hemicentrotus pulcherrimus, Stomopneustes variolaris, and Strongylocentrotus nudus, and as a smaller form in Clypeaster japonicus and Temnopleurus hardwicki. These results suggest that the change in the molecular form of AchE induced a change in enzymatic activity that in turn may play a role in spicule elongation in sea urchin embryos.
Percolated microstructures for multi-modal transport enhancement in porous active materials
McKay, Ian Salmon; Yang, Sungwoo; Wang, Evelyn N.; Kim, Hyunho
2018-03-13
A method of forming a composite material for use in multi-modal transport includes providing three-dimensional graphene having hollow channels, enabling a polymer to wick into the hollow channels of the three-dimensional graphene, curing the polymer to form a cured three-dimensional graphene, adding an active material to the cured three-dimensional graphene to form a composite material, and removing the polymer from within the hollow channels. A composite material formed according to the method is also provided.
Formation of mutagens in cooked foods. I. Beef.
Spingarn, N E; Weisburger, J H
1979-09-01
Mutagens detectable by Salmonella typhimurium TA98, after activation by liver S-9 fraction, are formed when meat is cooked by frying, broiling and boiling. High levels of mutagenic activity are formed rapidly when frying, or more slowly during broiling. Formation of mutagens in boiled beef stock requires several days under reflux, but shows a strong concentration dependence. Time curves suggest that a period exists during which mutagens are not readily formed; however, after this period mutagen production is rapid. Hamburgers from commercial franchises were frequently mutagenically active.
Walking through Architectural Spaces: The Impact of Interior Forms on Human Brain Dynamics
Banaei, Maryam; Hatami, Javad; Yazdanfar, Abbas; Gramann, Klaus
2017-01-01
Neuroarchitecture uses neuroscientific tools to better understand architectural design and its impact on human perception and subjective experience. The form or shape of the built environment is fundamental to architectural design, but not many studies have shown the impact of different forms on the inhabitants’ emotions. This study investigated the neurophysiological correlates of different interior forms on the perceivers’ affective state and the accompanying brain activity. To understand the impact of naturalistic three-dimensional (3D) architectural forms, it is essential to perceive forms from different perspectives. We computed clusters of form features extracted from pictures of residential interiors and constructed exemplary 3D room models based on and representing different formal clusters. To investigate human brain activity during 3D perception of architectural spaces, we used a mobile brain/body imaging (MoBI) approach recording the electroencephalogram (EEG) of participants while they naturally walk through different interior forms in virtual reality (VR). The results revealed a strong impact of curvature geometries on activity in the anterior cingulate cortex (ACC). Theta band activity in ACC correlated with specific feature types (rs (14) = 0.525, p = 0.037) and geometry (rs (14) = −0.579, p = 0.019), providing evidence for a role of this structure in processing architectural features beyond their emotional impact. The posterior cingulate cortex and the occipital lobe were involved in the perception of different room perspectives during the stroll through the rooms. This study sheds new light on the use of mobile EEG and VR in architectural studies and provides the opportunity to study human brain dynamics in participants that actively explore and realistically experience architectural spaces. PMID:29033807
Walking through Architectural Spaces: The Impact of Interior Forms on Human Brain Dynamics.
Banaei, Maryam; Hatami, Javad; Yazdanfar, Abbas; Gramann, Klaus
2017-01-01
Neuroarchitecture uses neuroscientific tools to better understand architectural design and its impact on human perception and subjective experience. The form or shape of the built environment is fundamental to architectural design, but not many studies have shown the impact of different forms on the inhabitants' emotions. This study investigated the neurophysiological correlates of different interior forms on the perceivers' affective state and the accompanying brain activity. To understand the impact of naturalistic three-dimensional (3D) architectural forms, it is essential to perceive forms from different perspectives. We computed clusters of form features extracted from pictures of residential interiors and constructed exemplary 3D room models based on and representing different formal clusters. To investigate human brain activity during 3D perception of architectural spaces, we used a mobile brain/body imaging (MoBI) approach recording the electroencephalogram (EEG) of participants while they naturally walk through different interior forms in virtual reality (VR). The results revealed a strong impact of curvature geometries on activity in the anterior cingulate cortex (ACC). Theta band activity in ACC correlated with specific feature types ( r s (14) = 0.525, p = 0.037) and geometry ( r s (14) = -0.579, p = 0.019), providing evidence for a role of this structure in processing architectural features beyond their emotional impact. The posterior cingulate cortex and the occipital lobe were involved in the perception of different room perspectives during the stroll through the rooms. This study sheds new light on the use of mobile EEG and VR in architectural studies and provides the opportunity to study human brain dynamics in participants that actively explore and realistically experience architectural spaces.
Kromin, A A; Dvoenko, E E; Zenina, O Yu
2016-07-01
Reflection of the state of hunger in impulse activity of nose wing muscles and upper esophageal sphincter muscles was studied in chronic experiments on rabbits subjected to 24-h food deprivation in the absence of locomotion and during search behavior. In the absence of apparent behavioral activity, including sniffing, alai nasi muscles of hungry rabbits constantly generated bursts of action potentials synchronous with breathing, while upper esophageal sphincter muscles exhibited regular aperiodic low-amplitude impulse activity of tonic type. Latent form of food motivation was reflected in the structure of temporal organization of impulse activity of alai nasi muscles in the form of bimodal distribution of interpulse intervals and in temporal structure of impulse activity of upper esophageal sphincter muscles in the form of monomodal distribution. The latent form of food motivation was manifested in the structure of temporal organization of periods of the action potentials burst-like rhythm, generated by alai nasi muscles, in the form of monomodal distribution, characterized by a high degree of dispersion of respiratory cycle periods. In the absence of physical activity hungry animals sporadically exhibited sniffing activity, manifested in the change from the burst-like impulse activity of alai nasi muscles to the single-burst activity type with bimodal distribution of interpulse intervals and monomodal distribution of the burst-like action potentials rhythm periods, the maximum of which was shifted towards lower values, which was the cause of increased respiratory rate. At the same time, the monomodal temporal structure of impulse activity of the upper esophageal sphincter muscles was not changed. With increasing food motivation in the process of search behavior temporal structure of periods of the burst-like action potentials rhythm, generated by alai nasi muscles, became similar to that observed during sniffing, not accompanied by animal's locomotion, which is typical for the increased respiratory rhythm frequency. Increased hunger motivation was reflected in the temporal structure of impulse activity of upper esophageal sphincter muscles in the form of a shift to lower values of the maximum of monomodal distribution of interpulse intervals on the histogram, resulting in higher impulse activity frequency. The simultaneous increase in the frequency of action potentials bursts generation by alai nasi muscles and regular impulse activity of upper esophageal sphincter muscles is a reliable criterion for enhanced food motivation during search behavior in rabbits.
Characterization of the methemoglobin forming metabolites of benzocaine and lidocaine.
Hartman, Neil; Zhou, Hongfei; Mao, Jinzhe; Mans, Daniel; Boyne, Michael; Patel, Vikram; Colatsky, Thomas
2017-05-01
1. Topical anesthesia with benzocaine or lidocaine occasionally causes methemoglobinemia, an uncommon but potentially fatal disorder where the blood has a reduced ability to transport oxygen. Previous in vitro studies using human whole blood have shown that benzocaine causes more methemoglobin (MetHb) formation than lidocaine, and that both compounds require metabolic transformation to form the MetHb producing species. In the current investigation, the active species of benzocaine forming the MetHb was investigated. 2. HPLC analysis of benzocaine samples incubated with human hepatic S9 showed the formation of a peak with the same UV spectrum and retention time as benzocaine hydroxylamine (BenzNOH). To confirm the activity of BenzNOH, MetHb production following exposure to the compound was determined in whole human blood using an Avoximeter 4000 CO-oximeter. 3. BenzNOH produced MetHb in a concentration dependent manner without the need for metabolic activation. Benzocaine in the presence of metabolic activation required a concentration of 500 μM to produce a similar degree of MetHb formation as 20 μM BenzNOH without activation. Previous work suggested that two metabolites of lidocaine may also form MetHb; N-hydroxyxylidine and 4-hydroxyxylidine. Of these two metabolites 4-hydroxyxylidine produced the most MetHb in whole blood in vitro in the absence of metabolic activation, however BenzNOH produced up to 14.2 times more MetHb than 4-hydroxyxylidine at a similar concentration. 4. These results suggest that the ability of benzocaine to form MetHb is likely to be mediated through its hydroxylamine metabolite and that this metabolite is inherently more active than the potentially MetHb-forming metabolites of lidocaine.
Kim, Paul Y.; Vu, Trang T.; Leslie, Beverly A.; Stafford, Alan R.; Fredenburgh, James C.; Weitz, Jeffrey I.
2014-01-01
Fibrin (Fn) clots formed from γ′-fibrinogen (γ′-Fg), a variant with an elongated γ-chain, are resistant to lysis when compared with clots formed from the predominant γA-Fg, a finding previously attributed to differences in clot structure due to delayed thrombin-mediated fibrinopeptide (FP) B release or impaired cross-linking by factor XIIIa. We investigated whether slower lysis of γ′-Fn reflects delayed plasminogen (Pg) binding and/or activation by tissue plasminogen activator (tPA), reduced plasmin-mediated proteolysis of γ′-Fn, and/or altered cross-linking. Clots formed from γ′-Fg lysed more slowly than those formed from γA-Fg when lysis was initiated with tPA/Pg when FPA and FPB were both released, but not when lysis was initiated with plasmin, or when only FPA was released. Pg bound to γ′-Fn with an association rate constant 22% lower than that to γA-Fn, and the lag time for initiation of Pg activation by tPA was longer with γ′-Fn than with γA-Fn. Once initiated, however, Pg activation kinetics were similar. Factor XIIIa had similar effects on clots formed from both Fg isoforms. Therefore, slower lysis of γ′-Fn clots reflects delayed FPB release, which results in delayed binding and activation of Pg. When clots were formed from Fg mixtures containing more than 20% γ′-Fg, the upper limit of the normal level, the delay in lysis was magnified. These data suggest that circulating levels of γ′-Fg modulate the susceptibility of clots to lysis by slowing Pg activation by tPA and provide another example of the intimate connections between coagulation and fibrinolysis. PMID:25128532
Anatomical Substrates of Visual and Auditory Miniature Second-language Learning
Newman-Norlund, Roger D.; Frey, Scott H.; Petitto, Laura-Ann; Grafton, Scott T.
2007-01-01
Longitudinal changes in brain activity during second language (L2) acquisition of a miniature finite-state grammar, named Wernickese, were identified with functional magnetic resonance imaging (fMRI). Participants learned either a visual sign language form or an auditory-verbal form to equivalent proficiency levels. Brain activity during sentence comprehension while hearing/viewing stimuli was assessed at low, medium, and high levels of proficiency in three separate fMRI sessions. Activation in the left inferior frontal gyrus (Broca’s area) correlated positively with improving L2 proficiency, whereas activity in the right-hemisphere (RH) homologue was negatively correlated for both auditory and visual forms of the language. Activity in sequence learning areas including the premotor cortex and putamen also correlated with L2 proficiency. Modality-specific differences in the blood oxygenation level-dependent signal accompanying L2 acquisition were localized to the planum temporale (PT). Participants learning the auditory form exhibited decreasing reliance on bilateral PT sites across sessions. In the visual form, bilateral PT sites increased in activity between Session 1 and Session 2, then decreased in left PT activity from Session 2 to Session 3. Comparison of L2 laterality (as compared to L1 laterality) in auditory and visual groups failed to demonstrate greater RH lateralization for the visual versus auditory L2. These data establish a common role for Broca’s area in language acquisition irrespective of the perceptual form of the language and suggest that L2s are processed similar to first languages even when learned after the ‘‘critical period.’’ The right frontal cortex was not preferentially recruited by visual language after accounting for phonetic/structural complexity and performance. PMID:17129186
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Priming a perspective in Spanish monolingual children: the use of syntactic alternatives.
Gámez, Perla B; Shimpi, Priya M; Waterfall, Heidi R; Huttenlocher, Janellen
2009-03-01
We used a syntactic priming paradigm to show priming effects for active and passive forms in monolingual Spanish-speaking four- and five-year-olds. In a baseline experiment, we examined children's use of the fue-passive form and found it was virtually non-existent in their speech, although they produced important elements of the form. Children used a more frequent Spanish passive form, the subjectless/se-passive. In a priming experiment, we presented children with drawings described using either active or fue-passive sentences. Children then described novel drawings. Priming was induced for active and passive forms; however, children did not produce the fue-passive provided for them. Instead, children used the subjectless/se-passive and what we term the function-passive, which like the fue-passive, emphasize the patient of the action. We argue that children's use of different passive forms suggests they are sensitive to experimenter's input as it relates to scene interpretation and to syntax.
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..., Case Assistance Form; (Form DHS-7001), Online Ombudsman Form DHS-7001 AGENCY: Office of the Citizenship... System'' to ``Online Ombudsman Form DHS-7001''. The instructions have been updated to reflect the... Services Ombudsman--001 Virtual Ombudsman System (March 2010) to reflect the name change to Online...
Massive Star Formation Viewed through Extragalactic-Tinted Glasses
NASA Astrophysics Data System (ADS)
Willis, Sarah; Marengo, M.; Smith, H. A.; Allen, L.
2014-01-01
Massive Galactic star forming regions are the local analogs to the luminous star forming regions that dominate the emission from star forming galaxies. Their proximity to us enables the characterization of the full range of stellar masses that form in these more massive environments, improving our understanding of star formation tracers used in extragalactic studies. We have surveyed a sample of massive star forming regions with a range of morphologies and luminosities to probe the star formation activity in a variety of environments. We have used Spitzer IRAC and deep ground based J, H, Ks observations to characterize the Young Stellar Object (YSO) content of 6 massive star forming regions. These YSOs provide insight into the rate and efficiency of star formation within these regions, and enable comparison with nearby, low mass star forming regions as well as extreme cases of Galactic star formation including ‘mini-starburst’ regions. In addition, we have conducted an in-depth analysis of NGC 6334 to investigate how the star formation activity varies within an individual star forming region, using Herschel data in the far-infrared to probe the earliest stages of the ongoing star formation activity.
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2010-11-02
...-89D), Coastal Structures Form; FEMA Form 086-0-27E (formerly FEMA Form 81-89E), Alluvial Fan Flooding... Fan Flooding Form. Abstract: The certification forms are designed to assist requesters in gathering...
NASA Astrophysics Data System (ADS)
Badave, Kirti; Patil, Yogesh; Gonnade, Rajesh; Srinivas, Darbha; Dasgupta, Rajan; Khan, Ayesha; Rane, Sandhya
2011-12-01
Compound 1 [1-imino (acetyl hydrazino)-Vitamin K 3], displays valence tautomerically related electronic isomers as Form I and Form II. Form I exhibits 2D packing fragment with 1D ribbon chains of N-H⋯O hydrogen bonds and shows EPR silent features. While Form II is EPR active and exhibits biradical nature with double quantum transitions at g = 2.0040. 1H NMR of compound 2, [1-imino (hydrazino carboxylate)-Vitamin K 3] and Form II exhibit π delocalization via resonance assisted H-bonding [RAHB] effect compared to Form I. Molecular interactions in Form I and II are visualized by DSC. The electronic structures of compounds 1 and 2 have been correlated to their API values by measuring anticancer activities, mitochondrial potentials and DNA shearing patterns. Form II and compound 2 indicate mitochondria mediated apoptosis (˜75% cell death) while Form I causes 35% cell death.
Rojnik, Matija; Jevnikar, Zala R; Doljak, Bojan; Turk, Samo; Zidar, Nace; Kos, Janko
2012-10-01
Cathepsin H is a unique member of the cysteine cathepsins that acts primarily as an aminopeptidase. Like other cysteine cathepsins, it is synthesized as an inactive precursor and activated by proteolytic removal of its propeptide. Here we demonstrate that, in human cells, the processing of the propeptide is an autocatalytic, multistep process proceeding from an inactive 41kDa pro-form, through a 30kDa intermediate form, to the 28kDa mature form. Tyr87P and Gly90P were identified as the two major endopeptidase cleavage sites, converting the 30kDa form into the mature 28kDa form. The level of processing differs significantly in different human cell lines. In monocyte-derived macrophages U937 and prostate cancer cells PC-3, the 28kDa form is predominant, whereas in osteoblasts HOS the processing from the 30kDa form to the 28kDa form is significantly lower. The aminopeptidase activity of the enzyme and its subcellular localization are independent of the product, however the 30kDa form was not secreted in HOS cells. The activity of the resulting cathepsin H in U937 cells was significantly lower than that in HOS cells, presumably due to the high levels of endogenous cysteine protease inhibitor cystatin F present specifically in this cell line. These results provide an insight into the dependence of human cathepsin H processing and regulation on cell type. Copyright © 2012 Elsevier GmbH. All rights reserved.
Carroni, Marta; Franke, Kamila B; Maurer, Michael; Jäger, Jasmin; Hantke, Ingo; Gloge, Felix; Linder, Daniela; Gremer, Sebastian; Turgay, Kürşad; Bukau, Bernd; Mogk, Axel
2017-11-22
Ring-forming AAA+ chaperones exert ATP-fueled substrate unfolding by threading through a central pore. This activity is potentially harmful requiring mechanisms for tight repression and substrate-specific activation. The AAA+ chaperone ClpC with the peptidase ClpP forms a bacterial protease essential to virulence and stress resistance. The adaptor MecA activates ClpC by targeting substrates and stimulating ClpC ATPase activity. We show how ClpC is repressed in its ground state by determining ClpC cryo-EM structures with and without MecA. ClpC forms large two-helical assemblies that associate via head-to-head contacts between coiled-coil middle domains (MDs). MecA converts this resting state to an active planar ring structure by binding to MD interaction sites. Loss of ClpC repression in MD mutants causes constitutive activation and severe cellular toxicity. These findings unravel an unexpected regulatory concept executed by coiled-coil MDs to tightly control AAA+ chaperone activity.
ERIC Educational Resources Information Center
Schwendimann, Beat A.; Linn, Marcia C.
2016-01-01
Concept map activities often lack a subsequent revision step that facilitates knowledge integration. This study compares two collaborative critique activities using a Knowledge Integration Map (KIM), a form of concept map. Four classes of high school biology students (n?=?81) using an online inquiry-based learning unit on evolution were assigned…
Barnard, J P; Pedersen, P L
1994-08-15
In the presence of glucose and ample oxygen, insect form African trypanosomes release pyruvate more than 100-fold more slowly than do bloodstream forms. This rate decrease could not be accounted for simply by an increased mitochondrial pyruvate oxidation rate as inhibiting mitochondrial respiration increases pyruvate efflux to rates only 2-3% of that observed for bloodstream form trypanosomes. Alternatively, decreased pyruvate efflux from insect form trypanosomes could not be accounted for by decreased pyruvate transporter activity, which, surprisingly, was nearly as high in insect form trypanosomes as reported by us earlier for bloodstream forms (J.P. Barnard, B. Reynafarje, and P.L. Pedersen (1993) J. Biol. Chem. 268, 3654-3661). Rather, the low pyruvate efflux rate appears to be due primarily to reduced levels of the enzyme pyruvate kinase, which, in contrast to conclusions of an earlier study, is readily detected in insect form trypanosomes in the absence of added activators at an activity level about 4% of that found in bloodstream forms. Insect form pyruvate kinase seems to be located in the cytosol and exhibits kinetic profiles and constants nearly identical to those reported by us earlier for the bloodstream form enzyme (J.P. Barnard, and P.L. Pedersen (1988) Mol. Biochem. Parasitol. 31, 141-148). It is suggested that the reduced levels of pyruvate kinase, and hence the reduced pyruvate efflux rates, in insect form trypanosomes result from down regulation of the gene encoding the cytosolic enzyme.
Infrared light sources with semimetal electron injection
Kurtz, Steven R.; Biefeld, Robert M.; Allerman, Andrew A.
1999-01-01
An infrared light source is disclosed that comprises a layered semiconductor active region having a semimetal region and at least one quantum-well layer. The semimetal region, formed at an interface between a GaAsSb or GalnSb layer and an InAsSb layer, provides electrons and holes to the quantum-well layer to generate infrared light at a predetermined wavelength in the range of 2-6 .mu.m. Embodiments of the invention can be formed as electrically-activated light-emitting diodes (LEDs) or lasers, and as optically-pumped lasers. Since the active region is unipolar, multiple active regions can be stacked to form a broadband or multiple-wavelength infrared light source.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Xu, Wu; Canfield, Nathan L.; Zhang, Ji-Guang
Methods for making composite anodes, such as macroporous composite anodes, are disclosed. Embodiments of the methods may include forming a tape from a slurry including a substrate metal precursor, an anode active material, a pore-forming agent, a binder, and a solvent. A laminated structure may be prepared from the tape and sintered to produce a porous structure, such as a macroporous structure. The macroporous structure may be heated to reduce a substrate metal precursor and/or anode active material. Macroporous composite anodes formed by some embodiments of the disclosed methods comprise a porous metal and an anode active material, wherein themore » anode active material is both externally and internally incorporated throughout and on the surface of the macroporous structure.« less
Devi, V. Kalpana; Baskar, R.; Varalakshmi, P.
1993-01-01
The effect of Musa paradisiaca stem kernel juice was investigated in experimental urolithiatic rats. Stone forming rats exhibited a significant elevation in the activities of two oxalate synthesizing enzymes - Glycollic acid oxidase and Lactate dehydrogenase. Deposition and excretion of stone forming constituents in kidney and urine were also increased in these rats. The enzyme activities and the level of crystalline components were lowered with the extract treatment. The extract also reduced the activities of urinary alkaline phosphatase, lactate dehydrogenase, r-glutamyl transferase, inorganic pyrophosphatase and β-glucuronidase in calculogenic rats. No appreciable changes were noticed with leucine amino peptidase activity in treated rats. PMID:22556626
International physical activity questionnaire: reliability and validity of the Turkish version.
Saglam, Melda; Arikan, Hulya; Savci, Sema; Inal-Ince, Deniz; Bosnak-Guclu, Meral; Karabulut, Erdem; Tokgozoglu, Lale
2010-08-01
Physical inactivity is a global problem which is related to many chronic health disorders. Physical activity scales which allow cross-cultural comparisons have been developed. The goal was to assess the reliability and validity of a Turkish version of the International Physical Activity Questionnaire (IPAQ). 1,097 university students (721 women, 376 men; ages 18-32) volunteered. Short and long forms of the IPAQ gave good agreement and comparable 1-wk. test-retest reliabilities. Caltrac accelerometer data were compared with IPAQ scores in 80 participants with good agreement for short and long forms. Turkish versions of the IPAQ short and long forms are reliable and valid in assessment of physical activity.
An activated form of ADAM10 is tumor selective and regulates cancer stem-like cells and tumor growth
Saha, Nayanendu; Eissman, Moritz F.; Xu, Kai; Llerena, Carmen; Kusebauch, Ulrike; Ding, Bi-Sen; Cao, Zhongwei; Rafii, Shahin; Ernst, Matthias; Scott, Andrew M.; Nikolov, Dimitar B.; Lackmann, Martin
2016-01-01
The transmembrane metalloprotease ADAM10 sheds a range of cell surface proteins, including ligands and receptors of the Notch, Eph, and erbB families, thereby activating signaling pathways critical for tumor initiation and maintenance. ADAM10 is thus a promising therapeutic target. Although widely expressed, its activity is normally tightly regulated. We now report prevalence of an active form of ADAM10 in tumors compared with normal tissues, in mouse models and humans, identified by our conformation-specific antibody mAb 8C7. Structure/function experiments indicate mAb 8C7 binds an active conformation dependent on disulfide isomerization and oxidative conditions, common in tumors. Moreover, this active ADAM10 form marks cancer stem-like cells with active Notch signaling, known to mediate chemoresistance. Importantly, specific targeting of active ADAM10 with 8C7 inhibits Notch activity and tumor growth in mouse models, particularly regrowth after chemotherapy. Our results indicate targeted inhibition of active ADAM10 as a potential therapy for ADAM10-dependent tumor development and drug resistance. PMID:27503072
Atapattu, Lakmali; Saha, Nayanendu; Chheang, Chanly; Eissman, Moritz F; Xu, Kai; Vail, Mary E; Hii, Linda; Llerena, Carmen; Liu, Zhanqi; Horvay, Katja; Abud, Helen E; Kusebauch, Ulrike; Moritz, Robert L; Ding, Bi-Sen; Cao, Zhongwei; Rafii, Shahin; Ernst, Matthias; Scott, Andrew M; Nikolov, Dimitar B; Lackmann, Martin; Janes, Peter W
2016-08-22
The transmembrane metalloprotease ADAM10 sheds a range of cell surface proteins, including ligands and receptors of the Notch, Eph, and erbB families, thereby activating signaling pathways critical for tumor initiation and maintenance. ADAM10 is thus a promising therapeutic target. Although widely expressed, its activity is normally tightly regulated. We now report prevalence of an active form of ADAM10 in tumors compared with normal tissues, in mouse models and humans, identified by our conformation-specific antibody mAb 8C7. Structure/function experiments indicate mAb 8C7 binds an active conformation dependent on disulfide isomerization and oxidative conditions, common in tumors. Moreover, this active ADAM10 form marks cancer stem-like cells with active Notch signaling, known to mediate chemoresistance. Importantly, specific targeting of active ADAM10 with 8C7 inhibits Notch activity and tumor growth in mouse models, particularly regrowth after chemotherapy. Our results indicate targeted inhibition of active ADAM10 as a potential therapy for ADAM10-dependent tumor development and drug resistance. © 2016 Atapattu et al.
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Signal Transduction by a Fungal NOD-Like Receptor Based on Propagation of a Prion Amyloid Fold
Daskalov, Asen; Habenstein, Birgit; Martinez, Denis; Debets, Alfons J. M.; Sabaté, Raimon; Loquet, Antoine; Saupe, Sven J.
2015-01-01
In the fungus Podospora anserina, the [Het-s] prion induces programmed cell death by activating the HET-S pore-forming protein. The HET-s β-solenoid prion fold serves as a template for converting the HET-S prion-forming domain into the same fold. This conversion, in turn, activates the HET-S pore-forming domain. The gene immediately adjacent to het-S encodes NWD2, a Nod-like receptor (NLR) with an N-terminal motif similar to the elementary repeat unit of the β-solenoid fold. NLRs are immune receptors controlling cell death and host defense processes in animals, plants and fungi. We have proposed that, analogously to [Het-s], NWD2 can activate the HET-S pore-forming protein by converting its prion-forming region into the β-solenoid fold. Here, we analyze the ability of NWD2 to induce formation of the β-solenoid prion fold. We show that artificial NWD2 variants induce formation of the [Het-s] prion, specifically in presence of their cognate ligands. The N-terminal motif is responsible for this prion induction, and mutations predicted to affect the β-solenoid fold abolish templating activity. In vitro, the N-terminal motif assembles into infectious prion amyloids that display a structure resembling the β-solenoid fold. In vivo, the assembled form of the NWD2 N-terminal region activates the HET-S pore-forming protein. This study documenting the role of the β-solenoid fold in fungal NLR function further highlights the general importance of amyloid and prion-like signaling in immunity-related cell fate pathways. PMID:25671553
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PROCESS FOR THE PRODUCTION OF AN ACTIVATED FORM OF UO$sub 2$
Polissar, M.J.
1957-09-24
A process for producing a highly active form of UO/sub 2/ characterized both by rapid oxidation in air and by rapid chlorination with CCl/sub 4/ vapor at an elevated temperature is reported. In accordance with the process, commercial UO/sub 2/, is subjected to a series of oxidation-reduction operations to produce a form of UC/sub 2/ of enhanced reactivity. By treatimg commercial UO/sub 2/ at a temperature between 335 and 485 deg C with methane, then briefly with an oxygen containing gas and followimg this by a second treatment with a methane containing gas, the original relatively stable charge of UO/sub 2/ will be transformed into an active form of UO/sub 2/.
Gasdermins: Effectors of Pyroptosis.
Kovacs, Stephen B; Miao, Edward A
2017-09-01
Pyroptosis is a form of lytic programmed cell death initiated by inflammasomes, which detect cytosolic contamination or perturbation. This drives activation of caspase-1 or caspase-11/4/5, which cleave gasdermin D, separating its N-terminal pore-forming domain (PFD) from the C-terminal repressor domain (RD). The PFD oligomerizes to form large pores in the membrane that drive swelling and membrane rupture. Gasdermin D is one of six (in humans) gasdermin family members; several other gasdermins have also been shown to form pores that cause pyroptosis after cleavage to activate their PFDs. One of these, gasdermin E, is activated by caspase-3 cleavage. We review our current understanding of pyroptosis as well as current knowledge of the gasdermin family. Copyright © 2017 Elsevier Ltd. All rights reserved.
Yan, Kelly; Merritt, Hanne; Crawford, Kenneth; Pardee, Gwynn; Cheng, Jan Marie; Widger, Stephania; Hekmat-Nejad, Mohammad; Zaror, Isabel; Sim, Janet
2015-06-01
Extracellular signal-regulated kinase 2 (ERK2) is a serine/threonine protein kinase involved in many cellular programs, such as cell proliferation, differentiation, motility and programed cell-death. It is therefore considered an important target in the treatment of cancer. In an effort to support biochemical screening and small molecule drug discovery, we established a robust system to generate both inactive and active forms of ERK2 using insect expression system. We report here, for the first time, that inactive ERK2 can be expressed and purified with 100% homogeneity in the unphosphorylated form using insect system. This resulted in a significant 20-fold yield improvement compared to that previously reported using bacterial expression system. We also report a newly developed system to generate active ERK2 in insect cells through in vivo co-expression with a constitutively active MEK1 (S218D S222D). Isolated active ERK2 was confirmed to be doubly phosphorylated at the correct sites, T185 and Y187, in the activation loop of ERK2. Both ERK2 forms, inactive and active, were well characterized by biochemical activity assay for their kinase function. Inactive and active ERK2 were the two key reagents that enabled successful high through-put biochemical assay screen and structural drug discovery studies. Copyright © 2015 Elsevier Inc. All rights reserved.
Structure of the active form of human origin recognition complex and its ATPase motor module
Tocilj, Ante; On, Kin Fan; Yuan, Zuanning; Sun, Jingchuan; Elkayam, Elad; Li, Huilin; Stillman, Bruce; Joshua-Tor, Leemor
2017-01-01
Binding of the Origin Recognition Complex (ORC) to origins of replication marks the first step in the initiation of replication of the genome in all eukaryotic cells. Here, we report the structure of the active form of human ORC determined by X-ray crystallography and cryo-electron microscopy. The complex is composed of an ORC1/4/5 motor module lobe in an organization reminiscent of the DNA polymerase clamp loader complexes. A second lobe contains the ORC2/3 subunits. The complex is organized as a double-layered shallow corkscrew, with the AAA+ and AAA+-like domains forming one layer, and the winged-helix domains (WHDs) forming a top layer. CDC6 fits easily between ORC1 and ORC2, completing the ring and the DNA-binding channel, forming an additional ATP hydrolysis site. Analysis of the ATPase activity of the complex provides a basis for understanding ORC activity as well as molecular defects observed in Meier-Gorlin Syndrome mutations. DOI: http://dx.doi.org/10.7554/eLife.20818.001 PMID:28112645
Porcine CD38 exhibits prominent secondary NAD(+) cyclase activity.
Ting, Kai Yiu; Leung, Christina F P; Graeff, Richard M; Lee, Hon Cheung; Hao, Quan; Kotaka, Masayo
2016-03-01
Cyclic ADP-ribose (cADPR) mobilizes intracellular Ca(2+) stores and activates Ca(2+) influx to regulate a wide range of physiological processes. It is one of the products produced from the catalysis of NAD(+) by the multifunctional CD38/ADP-ribosyl cyclase superfamily. After elimination of the nicotinamide ring by the enzyme, the reaction intermediate of NAD(+) can either be hydrolyzed to form linear ADPR or cyclized to form cADPR. We have previously shown that human CD38 exhibits a higher preference towards the hydrolysis of NAD(+) to form linear ADPR while Aplysia ADP-ribosyl cyclase prefers cyclizing NAD(+) to form cADPR. In this study, we characterized the enzymatic properties of porcine CD38 and revealed that it has a prominent secondary NAD(+) cyclase activity producing cADPR. We also determined the X-ray crystallographic structures of porcine CD38 and were able to observe conformational flexibility at the base of the active site of the enzyme which allow the NAD(+) reaction intermediate to adopt conformations resulting in both hydrolysis and cyclization forming linear ADPR and cADPR respectively. © 2016 The Protein Society.
A novel proteolytic processing of prolysyl oxidase.
Atsawasuwan, Phimon; Mochida, Yoshiyuki; Katafuchi, Michitsuna; Tokutomi, Kentaro; Mocanu, Viorel; Parker, Carol E; Yamauchi, Mitsuo
2011-01-01
Lysyl oxidase (LOX) is an amine oxidase that is critical for the stability of connective tissues. The secreted proLOX is enzymatically quiescent and is activated through proteolytic cleavage between residues Gly(162) and Asp(163) (residue numbers according to the mouse LOX) by bone morphogenetic protein (BMP)-1 gene products. Here we report a novel processing of proLOX identified in vitro and in vivo. Two forms of mature LOX were identified and characterized by their immunoreactivity to specific antibodies, amine oxidase activity, and mass spectrometry. One form was identified as a well-characterized BMP-1 processed LOX protein. Another was found to be a truncated form of LOX resulting from the cleavage at the carboxy terminus of Arg(192). The truncated form of LOX still appeared to retain amine oxidase activity. The results from the proLOX gene deletion and mutation experiments indicated that the processing occurs independent of the cleavage of proLOX by BMP-1 gene products and likely requires the presence of LOX propeptide. These results indicate that proLOX could be processed by two different mechanisms producing two forms of active LOX.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Mendelsohn, M.E.; Yan Zhu; O'Neill, S.
Thrombin plays a critical role in platelet activation, hemostasis, and thrombosis. Cellular activation by thrombin leads to the phosphorylation of multiple proteins, most of which are unidentified. The authors have characterized several 29-kDa proteins that are rapidly phosphorylated following exposure of intact human platelets to thrombin. A murine monoclonal antibody raised to an unidentified estrogen receptor-related 29-kDa protein selectively recognized these proteins as well as a more basic, unphosphorylated 27-kDa protein. Cellular activation by thrombin led to a marked shift in the proportion of protein from the 27-kDa unphosphorylated form to the 29-kDa phosphoprotein species. Using this antibody, they isolatedmore » and sequenced a human cDNA clone encoding a protein that was identical to the mammalian 27-kDa heat shock protein (HSP27), a protein of uncertain function that is known to be phosphorylated to several forms and to be transcriptionally induced by estrogen. The 29-kDa proteins were confirmed to be phosphorylated forms of HSP27 by immunoprecipitation studies. Thus, the estrogen receptor-related protein is HSP27, and the three major 20-kDa proteins phosphorylated in thrombin-activated platelets are forms of HSP27. These data suggest a role for HSP27 in the signal transduction events of platelet activation.« less
Johansson, T; Nyman, P O
1993-01-01
The basidiomycete Trametes versicolor is a white-rot fungus and a potent degrader of lignin. The development of extracellular enzyme activities in the fungal culture under physiological conditions of secondary metabolism was investigated. Using the culture medium as starting material a large number of peroxidase forms were purified by the use of chromatographic techniques. Sixteen forms of lignin peroxidase and five forms of manganese(II) peroxidase were separated and the majority of these enzymes was characterized with respect to isoelectric point, molecular mass, and specific enzyme activity. The manganese(II) peroxidases showed a lower isoelectric point (pI 3.2-2.9) and a slightly higher molecular mass (44-45 kDa) than the lignin peroxidases (pI 3.7-3.1, and 41-43 kDa). Specific enzyme activities for the forms of lignin peroxidase, using veratryl alcohol as the substrate, were found to differ considerably. Certain differences in the specific enzyme activity were also observed among the forms of manganese(II) peroxidase. A multitude of peroxidase forms has previously been encountered in another white-rot fungus, Phanerochaete chrysosporium. The discovery that it also occurs in T. versicolor would suggest that this multiplicity could be a common feature among white-rot fungi and may be essential for the biodegradation of lignin.
Understanding the internal states of others by listening to action verbs.
Di Cesare, G; Fasano, F; Errante, A; Marchi, M; Rizzolatti, G
2016-08-01
The internal state of others can be understood observing their actions or listening to their voice. While the neural bases of action style (vitality forms) have been investigated, there is no information on how we recognize others' internal state by listening to their speech. Here, using fMRI technique, we investigated the neural correlates of auditory vitality forms while participants listened to action verbs in three different conditions: human voice pronouncing the verbs in a rude and gentle way, robot voice pronouncing the same verbs without vitality forms, and a scrambled version of the same verbs pronounced by human voice. In agreement with previous studies on vitality forms encoding, we found specific activation of the central part of insula during listening to human voice conveying specific vitality forms. In addition, when listening both to human and robot voices there was an activation of the posterior part of the left inferior frontal gyrus and of the parieto-premotor circuit typically described to be activated during observation and execution of arm actions. Finally, the superior temporal gyrus was activated bilaterally in all three conditions. We conclude that, the central part of insula is a key region for vitality forms processing allowing the understanding of the vitality forms regardless of the modality by which they are conveyed. Copyright © 2016. Published by Elsevier Ltd.
Di Giorgio, C; Delmas, F; Ollivier, E; Elias, R; Balansard, G; Timon-David, P
2004-01-01
Harmane, harmine, and harmaline were investigated for their in vitro antileishmanial activity toward parasites of the species Leishmania infantum. Harmane and Harmine displayed a moderate antiproliferative activity toward human monocytes and exerted a weak antileishmanial activity toward both the promastigote and the amastigote forms of the parasite. Their mechanism of action on the promastigote form of the parasite involved interactions with DNA metabolism leading to an accumulation of parasites in the S-G(2)M phases of the cell-cycle. Harmaline, at the contrary, was deprived from toxicity toward human cells and Leishmania promastigotes, however it exerted a strong antileishmanial activity toward the intracellular amastigote form of the parasite. This property was shown to partly result from the capacity of the molecule to prevent parasite internalization within macrophages by inhibiting Leishmania PKC activity.
Leishmanicidal activity of nine novel flavonoids from Delphinium staphisagria.
Ramírez-Macías, Inmaculada; Marín, Clotilde; Díaz, Jesús G; Rosales, María José; Gutiérrez-Sánchez, Ramón; Sánchez-Moreno, Manuel
2012-01-01
To evaluate the in vitro leishmanicidal activity of nine flavonoid derivatives from Delphinium staphisagria against L. infantum and L. braziliensis. The in vitro activity of compounds 1-9 was assayed on extracellular promastigote and axenic amastigote forms and on intracellular amastigote forms of the parasites. Infectivity and cytotoxicity tests were carried on J774.2 macrophage cells using Glucantime as the reference drug. The mechanisms of action were analysed performing metabolite excretion and transmission electronic microscope ultrastructural alteration studies. Nine flavonoids showed leishmanicidal activity against promastigote as well as amastigote forms of Leishmania infantum and L. braziliensis. These compounds were nontoxic to mammalian cells and were effective at similar concentrations up to or lower than that of the reference drug (Glucantime). The results showed that 2(″)-acetylpetiolaroside (compound 8) was clearly the most active. This study has demonstrated that flavonoid derivatives are active against L. infantum and L. braziliensis.
Activation of human acid sphingomyelinase through modification or deletion of C-terminal cysteine.
Qiu, Huawei; Edmunds, Tim; Baker-Malcolm, Jennifer; Karey, Kenneth P; Estes, Scott; Schwarz, Cordula; Hughes, Heather; Van Patten, Scott M
2003-08-29
One form of Niemann-Pick disease is caused by a deficiency in the enzymatic activity of acid sphingomyelinase. During efforts to develop an enzyme replacement therapy based on a recombinant form of human acid sphingomyelinase (rhASM), purified preparations of the recombinant enzyme were found to have substantially increased specific activity if cell harvest media were stored for several weeks at -20 degrees C prior to purification. This increase in activity was found to correlate with the loss of the single free thiol on rhASM, suggesting the involvement of a cysteine residue. It was demonstrated that a variety of chemical modifications of the free cysteine on rhASM all result in substantial activation of the enzyme, and the modified cysteine responsible for this activation was shown to be the C-terminal residue (Cys629). Activation was also achieved by copper-promoted dimerization of rhASM (via cysteine) and by C-terminal truncation using carboxypeptidase Y. The role of the C-terminal cysteine in activation was confirmed by creating mutant forms of rhASM in which this residue was either deleted or replaced by a serine, with both forms having substantially higher specific activity than wild-type rhASM. These results indicate that purified rhASM can be activated in vitro by loss of the free thiol on the C-terminal cysteine via chemical modification, dimerization, or deletion of this amino acid residue. This method of activation is similar to the cysteine switch mechanism described previously for matrix metalloproteinases and could represent a means of posttranslational regulation of ASM activity in vivo.
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ERIC Educational Resources Information Center
Reinhardt, Claus H.; Rosen, Evelyne N.
2012-01-01
Many studies have demonstrated a superiority of active learning forms compared with traditional lecture. However, there is still debate as to what degree structuring is necessary with regard to high exam outcomes. Seventy-five students from a premedical school were randomly attributed to an active lecture group, a cooperative group, or a…
Effects of Three Forms of Reading-Based Output Activity on L2 Vocabulary Learning
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Rassaei, Ehsan
2017-01-01
The current study investigated the effects of three forms of output activity on EFL learners' recognition and recall of second language (L2) vocabulary. To this end, three groups of learners of English as a foreign language (EFL) were instructed to employ the following three output activities after reading two narrative texts: (1) summarizing the…
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Kiesner, Eileen
In this colorfully illustrated kindergarten activity, students read (and re-read) "My Blue Suitcase" (Sharon Katz), as an introduction to traveling. The book uses all of the basic forms of transportation and forms the transportation lesson outline. The activity gives the students the task of learning about each mode of transportation:…
Okamura, Keisuke; Okuda, Tetsu; Shirai, Kazuyuki; Urata, Hidenori
2018-01-01
The aim of the present study was to establish a convenient clinically applicable assay method for chymase-dependent angiotensin II forming activity of circulating mononuclear leukocytes (CML), which was potentially a marker of tissue chymase activity. Using this method, association between CML chymase activity and clinical parameters was determined. Cardiovascular outpatients (n = 170) without taking antihypertensive medication were recruited. An ELISA for chymase-dependent angiotensin II-forming activity in CML was established using Nma /Dnp-modified angiotensin I. Logistic regression analysis revealed that age and male gender were significant independent determinants of the increased CML chymase activity. After adjustment by age and gender, the CML chymase activity was positively correlated with systolic blood pressure, pulse rate, and the brain natriuretic peptide level. The relation between blood pressure and CML chymase activity suggests that it might reflect that increased tissue chymase activity contributes to systemic high blood pressure and heart rate because plasma chymase is inactive due to inhibitory plasma inhibitors.
Sheehan, S M; Switzer, R L
1990-01-01
Western immunoblots and assays of Bacillus subtilis extracts showed that intracellular serine protease 1 is produced in a form larger than previously reported, appears not to have undergone N-terminal processing, and is active in the presence or absence of calcium. No evidence for an inactive precursor form of the protease was found. Images FIG. 1 PMID:2104610
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CASPASE-9 CARD:CORE DOMAIN INTERACTIONS REQUIRE A PROPERLY-FORMED ACTIVE SITE
Huber, Kristen L.; Serrano, Banyuhay P.; Hardy, Jeanne A.
2018-01-01
Caspase-9 is a critical factor in the initiation of apoptosis, and as a result is tightly regulated by a number of mechanisms. Caspase-9 contains a Caspase Activation and Recruitment Domain (CARD), which enables caspase-9 to form a tight interaction with the apoptosome, a heptameric activating platform. The caspase-9 CARD has been thought to be principally involved in recruitment to the apoptosome, but its roles outside this interaction have yet to be uncovered. In this work we show that the CARD is involved in physical interactions with the catalytic core of caspase-9 in the absence of the apoptosome; this interaction requires a properly formed caspase-9 active site. The active sites of caspases are composed of four extremely mobile loops. When the active-site loops are not properly ordered, the CARD and core domains of caspase-9 do not interact and behave independently, like loosely tethered beads. When the active-site loop bundle is properly ordered, the CARD domain interacts with the catalytic core, forming a single folding unit. Together these findings provide mechanistic insight into a new level of caspase-9 regulation, prompting speculation that the CARD may also play a role in the recruitment or recognition of substrate. PMID:29500231
Balthasar, Andrea J R; Huber, Walter; Weis, Susanne
2011-09-02
Homonym processing in German is of theoretical interest as homonyms specifically involve word form information. In a previous study (Weis et al., 2001), we found inferior parietal activation as a correlate of successfully finding a homonym from written stimuli. The present study tries to clarify the underlying mechanism and to examine to what extend the previous homonym effect is dependent on visual in contrast to auditory input modality. 18 healthy subjects were examined using an event-related functional magnetic resonance imaging paradigm. Participants had to find and articulate a homonym in relation to two spoken or written words. A semantic-lexical task - oral naming from two-word definitions - was used as a control condition. When comparing brain activation for solved homonym trials to both brain activation for unsolved homonyms and solved definition trials we obtained two activations patterns, which characterised both auditory and visual processing. Semantic-lexical processing was related to bilateral inferior frontal activation, whereas left inferior parietal activation was associated with finding the correct homonym. As the inferior parietal activation during successful access to the word form of a homonym was independent of input modality, it might be the substrate of access to word form knowledge. Copyright © 2011 Elsevier B.V. All rights reserved.
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Kim, Chaeeun; Park, Jun-Cheol; Choi, Sun Young; Kim, Yonghun; Seo, Seung-Young; Park, Tae-Eon; Kwon, Se-Hun; Cho, Byungjin; Ahn, Ji-Hoon
2018-04-01
2D layered materials with sensitive surfaces are promising materials for use in chemical sensing devices, owing to their extremely large surface-to-volume ratios. However, most chemical sensors based on 2D materials are used in the form of laterally defined active channels, in which the active area is limited to the actual device dimensions. Therefore, a novel approach for fabricating self-formed active-channel devices is proposed based on 2D semiconductor materials with very large surface areas, and their potential gas sensing ability is examined. First, the vertical growth phenomenon of SnS 2 nanocrystals is investigated with large surface area via metal-assisted growth using prepatterned metal electrodes, and then self-formed active-channel devices are suggested without additional pattering through the selective synthesis of SnS 2 nanosheets on prepatterned metal electrodes. The self-formed active-channel device exhibits extremely high response values (>2000% at 10 ppm) for NO 2 along with excellent NO 2 selectivity. Moreover, the NO 2 gas response of the gas sensing device with vertically self-formed SnS 2 nanosheets is more than two orders of magnitude higher than that of a similar exfoliated SnS 2 -based device. These results indicate that the facile device fabrication method would be applicable to various systems in which surface area plays an important role. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Mechanism of the Anticoagulant Activity of Thrombin Mutant W215A/E217A
DOE Office of Scientific and Technical Information (OSTI.GOV)
Gandhi, Prafull S.; Page, Michael J.; Chen, Zhiwei
2009-09-15
The thrombin mutant W215A/E217A (WE) is a potent anticoagulant both in vitro and in vivo. Previous x-ray structural studies have shown that WE assumes a partially collapsed conformation that is similar to the inactive E* form, which explains its drastically reduced activity toward substrate. Whether this collapsed conformation is genuine, rather than the result of crystal packing or the mutation introduced in the critical 215-217 {beta}-strand, and whether binding of thrombomodulin to exosite I can allosterically shift the E* form to the active E form to restore activity toward protein C are issues of considerable mechanistic importance to improve themore » design of an anticoagulant thrombin mutant for therapeutic applications. Here we present four crystal structures of WE in the human and murine forms that confirm the collapsed conformation reported previously under different experimental conditions and crystal packing. We also present structures of human and murine WE bound to exosite I with a fragment of the platelet receptor PAR1, which is unable to shift WE to the E form. These structural findings, along with kinetic and calorimetry data, indicate that WE is strongly stabilized in the E* form and explain why binding of ligands to exosite I has only a modest effect on the E*-E equilibrium for this mutant. The E* {yields} E transition requires the combined binding of thrombomodulin and protein C and restores activity of the mutant WE in the anticoagulant pathway.« less
Bermingham, Emma N.; Hesketh, John E.; Sinclair, Bruce R.; Koolaard, John P.; Roy, Nicole C.
2014-01-01
Selenium may play a beneficial role in multi-factorial illnesses with genetic and environmental linkages via epigenetic regulation in part via glutathione peroxidase (GPx) activity. A meta-analysis was undertaken to quantify the effects of dietary selenium supplementation on the activity of overall GPx activity in different tissues and animal species and to compare the effectiveness of different forms of dietary selenium. GPx activity response was affected by both the dose and form of selenium (p < 0.001). There were differences between tissues on the effects of selenium supplementation on GPx activity (p < 0.001); however, there was no evidence in the data of differences between animal species (p = 0.95). The interactions between dose and tissue, animal species and form were significant (p < 0.001). Tissues particularly sensitive to changes in selenium supply include red blood cells, kidney and muscle. The meta-analysis identified that for animal species selenium-enriched foods were more effective than selenomethionine at increasing GPx activity. PMID:25268836
[Active euthanasia in Colombia and assisted suicide in California].
Julesz, Máté
2016-01-31
The institution of active euthanasia has been legal in Colombia since 2015. In California, the regulation on physician-assisted suicide will come into effect on January 1, 2016. The legal institution of active euthanasia is not accepted under the law of the United States of America, however, physician-assisted suicide is accepted in an increasing number of member states. The related regulation in Oregon is imitated in other member states. In South America, Colombia is not the first country to legalize active euthanasia: active euthanasia has been legal in Uruguay since 1932. The North American legal tradition markedly differs from the South American one and both are incompatible with the Central European rule of law. In Hungary and in most European Union countries, solely the passive form of euthanasia is legal. In the Benelux countries, the active form of euthanasia is legal because the supranational law of the European Union does not prohibit it. Notwithstanding, European Union law does not prescribe legalization of either the active form of euthanasia, or the physician-assisted suicide.
Acosta de Pérez, O C; Koscinczuk, P; Teibler, P; Sánchez Negrette, M; Ruiz, R; Maruñak, S; Bogarín, G
1998-08-01
Hemorrhagic, oedema-forming activities and histopathological alterations in the mouse footpad induced by Bothrops and Crotalus snake venoms from Argentina. Hemorrhagic and oedema-forming activities of various Bothrops and Crotalus snake venoms from Argentina were studied, together with histological alterations in the mouse footpad. The highest oedema-forming activity was found in the venom of B. jararaca, followed by B. jararacussu, B. neuwiedii diporus, B. alternatus, and Crotalus durissus terrificus. Regarding hemorrhage, the highest activity was found in the venom of B. neuwiedii diporus, followed by B. jararacussu, B. alternatus, and B. jararaca. No hemorrhage was observed after injection of Crotalus durissus terrificus venom, in agreement with histological observations of injected footpads. Histological analysis revealed a conspicuous inflammatory reaction in the injected footpad, characterized by oedema and an inflammatory infiltrate rich in polymorphonuclear leucocytes. Necrotic blood vessels and dilated lymphatic vessels were observed after injection of B. jararaca and B. jararacussu venoms, and myonecrosis was evident in tissue of mice injected with B. alternatus and B. neuwiedii diporus.
An ELISA method detecting the active form of suPAR.
Zhou, Xiaolei; Xu, Mingming; Huang, Hailong; Mazar, Andrew; Iqbal, Zafar; Yuan, Cai; Huang, Mingdong
2016-11-01
Urokinase plasminogen activator receptor (uPAR) exists in a number of formats in human plasma, including soluble uPAR (suPAR) and uPAR fragments. We developed an ELISA method to detect specifically the active form suPAR, which binds to its natural ligand uPA. The intra CV and inter CV of this ELISA assay is 8.5% and 9.6% respectively, and the assay can recover 99.74% of added recombinant suPAR from 10% plasma. This assay is quite sensitive, capable of detecting down to 15pg/ml of suPAR, and can measure suPAR concentrations in the range of 0.031-8ng/ml with high linear relationship. Plasma samples from pregnant women were also measured for the active form of suPAR with this assay, giving an averaged level of 1.39ng/ml, slightly higher than the level of pooled plasma from healthy donors (0.96ng/ml). This study demonstrates the feasibility to measure the active form of suPAR, which will likely have value in clinical applications. Copyright © 2016. Published by Elsevier B.V.
How vertebrate and invertebrate visual pigments differ in their mechanism of photoactivation
Nakagawa, Masashi; Iwasa, Tatsuo; Kikkawa, Satoshi; Tsuda, Motoyuki; Ebrey, Thomas G.
1999-01-01
In vertebrate visual pigments, a glutamic acid serves as a negative counterion to the positively charged chromophore, a protonated Schiff base of retinal. When photoisomerization leads to the Schiff base deprotonating, the anionic glutamic acid becomes protonated, forming a neutral species that activates the visual cascade. We show that in octopus rhodopsin, the glutamic acid has no anionic counterpart. Thus, the “counterion” is already neutral, so no protonated form of an initially anionic group needs to be created to activate. This helps to explain another observation—that the active photoproduct of octopus rhodopsin can be formed without its Schiff base deprotonating. In this sense, the mechanism of light activation of octopus rhodopsin is simpler than for vertebrates, because it eliminates one of the steps required for vertebrate rhodopsins to achieve their activating state. PMID:10339563
Chen, Lei; Zhu, Yu; Yang, Danling; Zou, Rongfeng; Wu, Junchen; Tian, He
2014-01-01
In this report, antibacterial peptides1-3 were prepared with a spiropyran fluorescence probe. The probe exhibits a change in fluorescence when isomerized from a colorless spiro-form (spiropyran, Sp) to a colored open-form (merocyanine, Mc) under different chemical environments, which can be used to study the mechanism of antimicrobial activity. Peptides 1-3 exhibit a marked decrease in antimicrobial activity with increasing alkyl chain length. This is likely due to the Sp-Mc isomers in different polar environments forming different aggregate sizes in TBS, as demonstrated by time-dependent dynamic light scattering (DLS). Moreover, peptides 1-3 exhibited low cytotoxicity and hemolytic activity. These probe-modified peptides may provide a novel approach to study the effect of structural changes on antibacterial activity, thus facilitating the design of new antimicrobial agents to combat bacterial infection. PMID:25358905
Spatially resolved thermal desorption/ionization coupled with mass spectrometry
DOE Office of Scientific and Technical Information (OSTI.GOV)
Jesse, Stephen; Van Berkel, Gary J; Ovchinnikova, Olga S
2013-02-26
A system and method for sub-micron analysis of a chemical composition of a specimen are described. The method includes providing a specimen for evaluation and a thermal desorption probe, thermally desorbing an analyte from a target site of said specimen using the thermally active tip to form a gaseous analyte, ionizing the gaseous analyte to form an ionized analyte, and analyzing a chemical composition of the ionized analyte. The thermally desorbing step can include heating said thermally active tip to above 200.degree. C., and positioning the target site and the thermally active tip such that the heating step forms themore » gaseous analyte. The thermal desorption probe can include a thermally active tip extending from a cantilever body and an apex of the thermally active tip can have a radius of 250 nm or less.« less
Evidence for a latent form of protein phosphatase 1 associated with cardiac myofibrils.
Schlender, K K; Wang, W; Wilson, S E
1989-02-28
Detergent-purified myofibrils from bovine heart contained very little spontaneously active protein phosphatase 1 activity. Phosphatase 1, extracted from the myofibrils by freeze-thawing in the presence of 500 mM KCl, was markedly activated by cobalt/trypsin treatment. Myofibril phosphatase 1 was separated from phosphatase 2A by chromatography on heparin-Sepharose. The phosphatase 1 was isolated in a latent form. Pretreatment with trypsin released free catalytic subunit and increased activity about 25-fold. Addition of cobalt with the trypsin increased activity another 2-fold. The latent myofibril phosphatase 1 did not appear to be the same as previously characterized forms of protein phosphatase 1. We suggest that cardiac myofibril phosphatase 1 contains a unique inhibitory subunit which directs the enzyme to the myofibril and regulates the dephosphorylation of myofibril phosphoproteins.
Expanded RNA-binding activities of mammalian Argonaute 2
Tan, Grace S.; Garchow, Barry G.; Liu, Xuhang; Yeung, Jennifer; Morris, John P.; Cuellar, Trinna L.; McManus, Michael T.; Kiriakidou, Marianthi
2009-01-01
Mammalian Argonaute 2 (Ago2) protein associates with microRNAs (miRNAs) or small interfering RNAs (siRNAs) forming RNA-induced silencing complexes (RISCs/miRNPs). In the present work, we characterize the RNA-binding and nucleolytic activity of recombinant mouse Ago2. Our studies show that recombinant mouse Ago2 binds efficiently to miRNAs forming active RISC. Surprisingly, we find that recombinant mouse Ago2 forms active RISC using pre-miRNAs or long unstructured single stranded RNAs as guides. Furthermore, we demonstrate that, in vivo, endogenous human Ago2 binds directly to pre-miRNAs independently of Dicer, and that Ago2:pre-miRNA complexes are found both in the cytoplasm and in the nucleus of human cells. PMID:19808937
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Fathipour, Vahid; Khaki, Zohreh; Nassiri, Seyyed Mehdi
2018-01-01
Recently, matrix metalloproteinases (MMP), in particular the gelatinases MMP-2 and MMP-9, generally considered as tumor markers for clinical applications. A longitudinal 2-years follow-up survey was performed on dogs with cutaneous tumor. Serum samples were obtained from 22 dogs with different cutaneous tumors and 22 health dogs at the time of surgery and one month, three months and one year after surgery. Gelatin zymography, hematological and biochemical assessment were performed for all serum samples. The serum alkaline phosphatase activity in dogs with malignant tumors was significantly higher than that in dogs with benign tumors and control cases. Latent forms of MMP-2 and MMP-9 were detected in all of the tumor cases. Gelatin zymography showed active form of MMP-9 in 12 cases (three benign and nine malignant tumors) and active form of MMP-2 in one fibrosarcoma case. Serum activity of active-MMP-9 and total MMP-9 was significantly higher in dogs with cutaneous tumors than those in controls. Tumor cases had higher serum activity of active-MMP-9 rather than controls. MMPs and alkaline phosphatase activities in serum were decreased significantly after surgery. Only one case with perianal gland adenoma showed recurrence of tumor four months after surgery in which active form of MMP-9 had identified one month before recurrence. According to the findings, it will be useful to measure ALP, MMP-2 and MMP-9 activities in the serum of dogs with cutaneous tumor for determination of tumor behavior before surgical treatment.
Pozo, P; Valenzuela, M A; Melej, C; Zaldívar, M; Puente, J; Martínez, B; Gamonal, J
2005-06-01
The aim of this work was to improve the assessment of the periodontal disease status through measurements of extracellular matrix metalloproteinases (MMPs) and their tissular inhibitors (TIMPs) in the gingival crevicular fluid from patients diagnosed with chronic periodontitis. Gingival crevicular fluid samples from patients (n = 13) were taken from 60 sites initially, and from 51 and 41 sites, respectively, 3 and 6 months after scaling and root planing. Gingival crevicular fluid samples were also taken from healthy subjects (n = 11, 24 sites). The presence of MMP-9 and MMP-8 was assessed by zymography and immunowestern blotting, respectively. The actual MMP activity (gelatinase and collagenase) was measured using the fluorogenic substrate assay. TIMP-1 and -2 levels were measured by immunodot blot. The fluorogenic substrate assay determinations showed higher MMP activity in sites with probing depth > or = 4 mm, with significant reduction post-treatment. Gelatinase activity followed by zymography consisted mainly of MMP-9. A different pattern of MMP-8 in control and patient sites was found. Controls only showed species of a partially active form (69 kDa), whereas patient sites showed a high frequency of the active form (56 kDa), and in some cases the latent form (85 kDa) was also observed. The active form reduced its frequency in sites with probing depth > or = 4 mm. TIMP-1 and -2 levels in patients were significantly lower than in controls, and after treatment the recovery of TIMP-1 level similar to control was observed. Significant correlations between the severity of the periodontal disease and the actual MMP activity, the active form of MMP-8 and the low level of both TIMP-1 and TIMP-2 were found.
Stabilization of the E* Form Turns Thrombin into an Anticoagulant
DOE Office of Scientific and Technical Information (OSTI.GOV)
Bah, Alaji; Carrell, Christopher J.; Chen, Zhiwei
2009-07-31
Previous studies have shown that deletion of nine residues in the autolysis loop of thrombin produces a mutant with an anticoagulant propensity of potential clinical relevance, but the molecular origin of the effect has remained unresolved. The x-ray crystal structure of this mutant solved in the free form at 1.55 {angstrom} resolution reveals an inactive conformation that is practically identical (root mean square deviation of 0.154 {angstrom}) to the recently identified E* form. The side chain of Trp215 collapses into the active site by shifting >10 {angstrom} from its position in the active E form, and the oxyanion hole ismore » disrupted by a flip of the Glu192-Gly193 peptide bond. This finding confirms the existence of the inactive form E* in essentially the same incarnation as first identified in the structure of the thrombin mutant D102N. In addition, it demonstrates that the anticoagulant profile often caused by a mutation of the thrombin scaffold finds its likely molecular origin in the stabilization of the inactive E* form that is selectively shifted to the active E form upon thrombomodulin and protein C binding.« less
Process for forming a homogeneous oxide solid phase of catalytically active material
Perry, Dale L.; Russo, Richard E.; Mao, Xianglei
1995-01-01
A process is disclosed for forming a homogeneous oxide solid phase reaction product of catalytically active material comprising one or more alkali metals, one or more alkaline earth metals, and one or more Group VIII transition metals. The process comprises reacting together one or more alkali metal oxides and/or salts, one or more alkaline earth metal oxides and/or salts, one or more Group VIII transition metal oxides and/or salts, capable of forming a catalytically active reaction product, in the optional presence of an additional source of oxygen, using a laser beam to ablate from a target such metal compound reactants in the form of a vapor in a deposition chamber, resulting in the deposition, on a heated substrate in the chamber, of the desired oxide phase reaction product. The resulting product may be formed in variable, but reproducible, stoichiometric ratios. The homogeneous oxide solid phase product is useful as a catalyst, and can be produced in many physical forms, including thin films, particulate forms, coatings on catalyst support structures, and coatings on structures used in reaction apparatus in which the reaction product of the invention will serve as a catalyst.
Manhart, Carol M.; Ni, Xiaodan; White, Martin A.; Ortega, Joaquin; Surtees, Jennifer A.
2017-01-01
Crossing over between homologs is initiated in meiotic prophase by the formation of DNA double-strand breaks that occur throughout the genome. In the major interference-responsive crossover pathway in baker’s yeast, these breaks are resected to form 3' single-strand tails that participate in a homology search, ultimately forming double Holliday junctions (dHJs) that primarily include both homologs. These dHJs are resolved by endonuclease activity to form exclusively crossovers, which are critical for proper homolog segregation in Meiosis I. Recent genetic, biochemical, and molecular studies in yeast are consistent with the hypothesis of Mlh1-Mlh3 DNA mismatch repair complex acting as the major endonuclease activity that resolves dHJs into crossovers. However, the mechanism by which the Mlh1-Mlh3 endonuclease is activated is unknown. Here, we provide evidence that Mlh1-Mlh3 does not behave like a structure-specific endonuclease but forms polymers required to generate nicks in DNA. This conclusion is supported by DNA binding studies performed with different-sized substrates that contain or lack polymerization barriers and endonuclease assays performed with varying ratios of endonuclease-deficient and endonuclease-proficient Mlh1-Mlh3. In addition, Mlh1-Mlh3 can generate religatable double-strand breaks and form an active nucleoprotein complex that can nick DNA substrates in trans. Together these observations argue that Mlh1-Mlh3 may not act like a canonical, RuvC-like Holliday junction resolvase and support a novel model in which Mlh1-Mlh3 is loaded onto DNA to form an activated polymer that cleaves DNA. PMID:28453523
Manhart, Carol M; Ni, Xiaodan; White, Martin A; Ortega, Joaquin; Surtees, Jennifer A; Alani, Eric
2017-04-01
Crossing over between homologs is initiated in meiotic prophase by the formation of DNA double-strand breaks that occur throughout the genome. In the major interference-responsive crossover pathway in baker's yeast, these breaks are resected to form 3' single-strand tails that participate in a homology search, ultimately forming double Holliday junctions (dHJs) that primarily include both homologs. These dHJs are resolved by endonuclease activity to form exclusively crossovers, which are critical for proper homolog segregation in Meiosis I. Recent genetic, biochemical, and molecular studies in yeast are consistent with the hypothesis of Mlh1-Mlh3 DNA mismatch repair complex acting as the major endonuclease activity that resolves dHJs into crossovers. However, the mechanism by which the Mlh1-Mlh3 endonuclease is activated is unknown. Here, we provide evidence that Mlh1-Mlh3 does not behave like a structure-specific endonuclease but forms polymers required to generate nicks in DNA. This conclusion is supported by DNA binding studies performed with different-sized substrates that contain or lack polymerization barriers and endonuclease assays performed with varying ratios of endonuclease-deficient and endonuclease-proficient Mlh1-Mlh3. In addition, Mlh1-Mlh3 can generate religatable double-strand breaks and form an active nucleoprotein complex that can nick DNA substrates in trans. Together these observations argue that Mlh1-Mlh3 may not act like a canonical, RuvC-like Holliday junction resolvase and support a novel model in which Mlh1-Mlh3 is loaded onto DNA to form an activated polymer that cleaves DNA.
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Ticehurst, Martyn David; Marziano, Ivan
2015-06-01
This review seeks to offer a broad perspective that encompasses an understanding of the drug product attributes affected by active pharmaceutical ingredient (API) physical properties, their link to solid form selection and the role of particle engineering. While the crucial role of active pharmaceutical ingredient (API) solid form selection is universally acknowledged in the pharmaceutical industry, the value of increasing effort to understanding the link between solid form, API physical properties and drug product formulation and manufacture is now also being recognised. A truly holistic strategy for drug product development should focus on connecting solid form selection, particle engineering and formulation design to both exploit opportunities to access simpler manufacturing operations and prevent failures. Modelling and predictive tools that assist in establishing these links early in product development are discussed. In addition, the potential for differences between the ingoing API physical properties and those in the final product caused by drug product processing is considered. The focus of this review is on oral solid dosage forms and dry powder inhaler products for lung delivery. © 2015 Royal Pharmaceutical Society.
Disposable, Autonomic, Energy-Converting Ion Channel Sensor Materials
2018-07-02
variant forms well-defined pores _____________________________ 26 4.2. Another pore-forming peptide, Ceratotoxin A, displays alamethicin-like activity ...bilayer recordings to examine the activity of these compounds on the single-pore level. We plan to use modified CtxA for targeted cell killing...strongly dependent on entropy of activation . Tethering is one strategy towards achieving this goal. A manuscript regarding this work is currently in
Monolithically integrated solid state laser and waveguide using spin-on glass
Ashby, C.I.H.; Hohimer, J.P.; Neal, D.R.; Vawter, G.A.
1995-10-31
A monolithically integrated photonic circuit is disclosed combining a semiconductor source of excitation light with an optically active waveguide formed on the substrate. The optically active waveguide is preferably formed of a spin-on glass to which are added optically active materials which can enable lasing action, optical amplification, optical loss, or frequency conversion in the waveguide, depending upon the added material. 4 figs.
Michelle J. Serapiglia; Rakesh Minocha; Subhash C. Minocha
2008-01-01
We analyzed effects of nitrogen availability and form on growth rates, concentrations of polyamines and inorganic ions and glutamine synthetase activity in in-vitro-cultured red spruce (Picea rubens Sarg.) cells. Growth rates, concentrations of polyamines and glutamine synthetase activity declined when either the amount of nitrate or the total amount...
Drijvers, Linda; Mulder, Kimberley; Ernestus, Mirjam
2016-02-01
Reduced forms like yeshay for yesterday often occur in conversations. Previous behavioral research reported a processing advantage for full over reduced forms. The present study investigated whether this processing advantage is reflected in a modulation of alpha (8-12Hz) and gamma (30+Hz) band activity. In three electrophysiological experiments, participants listened to full and reduced forms in isolation (Experiment 1), sentence-final position (Experiment 2), or mid-sentence position (Experiment 3). Alpha power was larger in response to reduced forms than to full forms, but only in Experiments 1 and 2. We interpret these increases in alpha power as reflections of higher auditory cognitive load. In all experiments, gamma power only increased in response to full forms, which we interpret as showing that lexical activation spreads more quickly through the semantic network for full than for reduced forms. These results confirm a processing advantage for full forms, especially in non-medial sentence position. Copyright © 2016 Elsevier Inc. All rights reserved.
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Ostrakhovich, E A; Ilich-Stoianovich, O; Afanas'ev, I B
2001-01-01
Infrared pulse laser therapy was studied for its impact on the production of active forms of oxygen and nitrogen by neutrophils from patients with rheumatoid arthritis (RA). The authors determined the non-activated and PMA-activated production of superoxide anion-radical, peroxynitrite, peripheral neurophilic NAD.PH-oxidase and superoxide dismutase activities, and the red blood cell concentrations of reduced glutathione. Before therapy, non-activation RA neurophilic production of superoxide was much higher than in donors. Laser therapy made this parameter normal. Similarly, neutrophilic peroxynitrite production (defined by dihydrorhodamine oxidation) in RA patients was 1.7 times higher than the normal values. IF-laser therapy decreased peroxynitrite production to the values observed in donors. It is important that the therapy caused increased SOD activity (that was lower in RA patients prior to therapy) up to apparently control values. Thus, IF-laser therapy has a certain antioxidative effect by increasing SOD activity in RA patients' blood cells and reducing the production of highly reactive oxygen and nitrogen forms.
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CTEPP DATA COLLECTION FORM 04: PARENT PRE-MONITORING QUESTIONNAIRE
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Expressing our internal states and understanding those of others.
Di Cesare, Giuseppe; Di Dio, Cinzia; Marchi, Massimo; Rizzolatti, Giacomo
2015-08-18
Vitality form is a term that describes the style with which motor actions are performed (e.g., rude, gentle, etc.). They represent one characterizing element of conscious and unconscious bodily communication. Despite their importance in interpersonal behavior, vitality forms have been, until now, virtually neglected in neuroscience. Here, using the functional MRI (fMRI) technique, we investigated the neural correlates of vitality forms in three different tasks: action observation, imagination, and execution. Conjunction analysis showed that, in all three tasks, there is a common, consistent activation of the dorsocentral sector of the insula. In addition, a common activation of the parietofrontal network, typically active during arm movements production, planning, and observation, was also found. We conclude that the dorsocentral part of the insula is a key element of the system that modulates the cortical motor activity, allowing individuals to express their internal states through action vitality forms. Recent monkey anatomical data show that the dorsocentral sector of the insula is, indeed, connected with the cortical circuit involved in the control of arm movements.
Ray, D S; Hines, J C
1995-01-01
Both prokaryotic and eukaryotic cells contain multiple forms of ribonuclease H, a ribonuclease that specifically degrades the RNA strand of RNA-DNA hybrids and which has been implicated in the processing of initiator RNAs and in the removal of RNA primers from Okazaki fragments. The Crithidia fasciculata RNH1 gene encodes an RNase H and was shown to be a single-copy gene in this diploid trypanosomatid. The RNH1 gene has been disrupted by targeted gene disruption using hygromycin or G418 drug-resistance cassettes. Major active forms of RNase H (38 and 45 kDa) were observed on activity gels of extracts of wild-type cells or cells in which one allele of RNH1 was disrupted. Both the 38 and 45 kDa activities were absent in extracts of cells in which both alleles of RNH1 were disrupted indicating that both forms of the C.fasciculata RNase H are encoded by the RNH1 gene. Images PMID:7630731
Weaver, J; Zhan, H; Pollack, S
1993-01-01
Iron first entering the reticulocyte is bound to ATP in the low MW cytosolic pool; some is also 'loosely bound' to haemoglobin, coeluting with haemoglobin from a molecular sieve column though not incorporated into haem. When haemolysate is mixed with ATP-Fe in vitro a similar high MW iron-containing complex is formed: the ATP-Fe interacts with a non-haemoglobin constituent of the haemolysate to form a high MW ATP-Fe complex in which the ratio of ATP:Fe (originally 6:1) is reversed, so that the complex contains more iron than ATP. The high MW ATP-Fe complex is formed even when ATP is in 150-fold molar excess and is formed without detectable hydrolysis of the ATP. The activity of haemolysate in forming the high MW ATP-Fe complex is not diminished by dialysis; all of the activity is recovered in the haemoglobin-containing fraction obtained from an Ultrogel AcA 44 column. The activity does not derive from haemoglobin since 85% of the activity is removed when haemoglobin is purified from haemolysate with DEAE-Sephadex. The chelatable iron pool of the cell probably includes both the high MW ATP-Fe complex and low MW ATP-Fe. Shunting of ATP-Fe to a high MW aggregate reduces the amount of iron present in the highly reactive low MW form and thus probably serves to limit the formation of cell damaging radicals.
Khoroshilova, Natalia; Popescu, Codrina; Münck, Eckard; Beinert, Helmut; Kiley, Patricia J.
1997-01-01
The transcription factor FNR (fumarate nitrate reduction) requires the presence of an iron-sulfur (Fe-S) cluster for its function as a global transcription regulator in Escherichia coli when oxygen becomes scarce. To define the oxidation state and type of Fe-S cluster present in the active form of FNR, we have studied anaerobically purified FNR with Mössbauer spectroscopy. Our data showed that this form of FNR contained a [4Fe-4S]2+ cluster (δ = 0.45 mm/s; ΔEQ = 1.22 mm/s) and that the [4Fe-4S]2+ cluster was rapidly destroyed on exposure of FNR to air. Under these conditions, the yellow–green active form of FNR turned deep red; analysis of sulfide indicated that 70% of the labile sulfide was still present, suggesting that the Fe-S cluster had been converted into a different form. Little [3Fe-4S] cluster was, however, detected by EPR. According to Mössbauer spectroscopy, the [4Fe-4S]2+ cluster was converted in about 60% yield to a [2Fe-2S]2+ cluster (δ = 0.28 mm/s; ΔEQ = 0.58 mm/s) following 17 min of exposure to air. The [2Fe-2S]2+ cluster form of FNR was much more stable to oxygen, but was unable to sustain biological activity (e.g., DNA binding). However, DNA binding and the absorption spectrum characteristic of the [4Fe-4S]2+ cluster could be largely restored from the [2Fe-2S]2+ form when Cys, Fe, DTT, and the NifS protein were added. It has yet to be determined whether the form of FNR containing the [2Fe-2S]2+ cluster has any biological significance, e.g., as an in vivo intermediate that is more rapidly converted to the active form than the apoprotein. PMID:9177174
Oroszlán, Gábor; Dani, Ráhel; Szilágyi, András; Závodszky, Péter; Thiel, Steffen; Gál, Péter; Dobó, József
2017-01-01
Serine proteases (SPs) are typically synthesized as precursors, termed proenzymes or zymogens, and the fully active form is produced via limited proteolysis by another protease or by autoactivation. The lectin pathway of the complement system is initiated by mannose-binding lectin (MBL)-associated SPs (MASP)-1, and MASP-2, which are known to be present as proenzymes in blood. The third SP of the lectin pathway, MASP-3, was recently shown to be the major activator, and the exclusive "resting blood" activator of profactor D, producing factor D, the initiator protease of the alternative pathway. Because only activated MASP-3 is capable of carrying out this cleavage, it was presumed that a significant fraction of MASP-3 must be present in the active form in resting blood. Here, we aimed to detect active MASP-3 in the blood by a more direct technique and to quantitate the active to zymogen ratio. First, MASPs were partially purified (enriched) from human plasma samples by affinity chromatography using immobilized MBL in the presence of inhibitors. Using this MASP pool, only the zymogen form of MASP-1 was detected by Western blot, whereas over 70% MASP-3 was in an activated form in the same samples. Furthermore, the active to zymogen ratio of MASP-3 showed little individual variation. It is enigmatic how MASP-3, which is not able to autoactivate, is present mostly as an active enzyme, whereas MASP-1, which has a potent autoactivation capability, is predominantly proenzymic in resting blood. In an attempt to explain this phenomenon, we modeled the basal level fluid-phase activation of lectin pathway proteases and their subsequent inactivation by C1 inhibitor and antithrombin using available and newly determined kinetic constants. The model can explain extensive MASP-3 activation only if we assume efficient intracomplex activation of MASP-3 by zymogen MASP-1. On the other hand, the model is in good agreement with the fact that MASP-1 and -2 are predominantly proenzymic and some of them is present in the form of inactive serpin-protease complexes. As an alternative hypothesis, MASP-3 activation by proprotein convertases is also discussed.
Gullies Old and New Near the Argyre Region
2015-01-28
This image from NASA Mars Reconnaissance Orbiter shows several seemingly active gullies and their associated fans near the Argyre region. As gullies form on slopes, sediment can become deposited to form lobe-shaped fans. Gullies can form with flowing water, but don't have to: sometimes they can be formed by the dry flow of sand and dust grains. This HiRISE image shows several seemingly active gullies and their associated fans. These particular gullies have many fine channels that are resolved nicely with a high resolution image. http://photojournal.jpl.nasa.gov/catalog/PIA19286
Espinosa, Ivette; Báez, Michel; Lobo, Evelyn; Martínez, Siomara; Gottschalk, Marcelo
2016-01-01
The aim of this study was to investigate the capacity of Streptococcus suis strains to form biofilms and to evaluate the antimicrobial activity of Penicillin G and N-acetylcystein (NAC) on both S. suis sessile and planktonic forms. Only non-typeable isolates of S. suis were correlated with a greater biofilm formation capacity. The MCI of Penicillin G and NAC required for inhibiting biofilm growth were higher than the required concentration for inhibiting planktonic growth. The combinations of NAC and Penicillin G showed a strong synergistic activity that inhibited biofilm formation and disrupted the pre-formed biofilm of S. suis.
Titov, V N; Oshchepkova, E V; Dmitriev, V A; Gushchina, O V; Shiriaeva, Iu K; Iashin, A Ia
2012-04-01
During millions years in all animals allantoine (oxidized by uricase uric acid) was catabolite of purines and ascorbic acid was an acceptor of active forms of oxygen. The proximal tubules of nephron reabsorbed the trace amounts of uric acid Then during phylogenesis the primates had a mutation of ascorbic acid gen minus. Later on occurred a second spontaneous mutation and uricase gen minus and uric acid became catabolites of purines. In absence of ascorbic acid synthesis ions of urates became a major capturers of active forms of oxygen and all uric acid as before underwent the reabsorption. Later the carriers were formed which began in epithelium of proximal tubules to secrete all uric acid into urine. At every incident of "littering" of intercellular medium with endogenic flogogens (impairment of biologic function of endoecology) under compensatory development of biologic reaction of inflammation the need in inactivation of active forms of oxygen increases. Hence later on in phylogenesis one more stage was formed--post secretory reabsorption of uric acid In the biologic reaction of inflammation epithelium of proximal tubules initiates retentional hyperiricosuria. The general antioxidant activity of human blood plasma in 60% is presented by urates' ions. The excretion of uric acid includes 4 stages: filtration, full reabsorption, secretion and post secretory reabsorption. In phylogenesis these stages formed in sequence. The mild hyperiricosuria is most frequently considered as a non-specific indicator of activation of biologic reaction of inflammation. The productive hyperiricosuria develops more infrequently under surplus of meat food and cytolysis syndrome (intensification of cell loss in vivo). Under concentration of uric acid more than 400 mkmol/l part of urates circulates in intercellular medium in the form of crystals. The microcrystals of uric acid (biologic "litter") initiate the syndrome of systemic inflammatory response as an endogenic flogogen--initiator of inflammation. The uric acid in the form of ion-capturers of active forms of oxygen is involved into in the formation of syndrome of compensatory anti-inflammatory defense. It may be assumed that simultaneously with post-secretory reabsorption of ions of urates in proximal tubules of nephron occurs intensification of philogenetically late post-secretory reabsorption of ions of sodium and activation of of biologic reaction of hydrodynamic and hydraulic pressure in local pool of intravascular medium i.e. arterial tension. The uric acid simultaneously participates in realization of biologic function of endoecology and adaptation, biologic reactions of excretion, inflammation and arterial tension.
Non-plaque-forming virions of Modified Vaccinia virus Ankara express viral genes.
Lülf, Anna-Theresa; Freudenstein, Astrid; Marr, Lisa; Sutter, Gerd; Volz, Asisa
2016-12-01
In cell culture infections with vaccinia virus the number of counted virus particles is substantially higher than the number of plaques obtained by titration. We found that standard vaccine preparations of recombinant Modified Vaccinia virus Ankara produce only about 20-30% plaque-forming virions in fully permissive cell cultures. To evaluate the biological activity of the non-plaque-forming particles, we generated recombinant viruses expressing fluorescent reporter proteins under transcriptional control of specific viral early and late promoters. Live cell imaging and automated counting by fluorescent microscopy indicated that virtually all virus particles can enter cells and switch on viral gene expression. Although most of the non-plaque-forming infections are arrested at the level of viral early gene expression, we detected activation of late viral transcription in 10-20% of single infected cells. Thus, non-plaque-forming particles are biologically active, and likely contribute to the immunogenicity of vaccinia virus vaccines. Copyright © 2016 Elsevier Inc. All rights reserved.
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Activation of methane by transition metal-substituted aluminophosphate molecular sieves
Iton, Lennox E.; Maroni, Victor A.
1991-01-01
Aluminophosphate molecular sieves substituted with cobalt, manganese or iron and having the AlPO.sub.4 -34 or AlPO.sub.4 -5, or related AlPO.sub.4 structure activate methane starting at approximately 350.degree. C. Between 400.degree. and 500.degree. C. and at methane pressures .ltoreq.1 atmosphere the rate of methane conversion increases steadily with typical conversion efficiencies at 500.degree. C. approaching 50% and selectivity to the production of C.sub.2+ hydrocarbons approaching 100%. The activation mechanism is based on reduction of the transition metal(III) form of the molecular sieve to the transition metal(II) form with accompanying oxidative dehydrogenation of the methane. Reoxidation of the - transition metal(II) form to the transition metal(III) form can be done either chemically (e.g., using O.sub.2) or electrochemically.
NASA Astrophysics Data System (ADS)
Zimakova, G. A.; Solonina, V. A.; Zelig, M. P.
2017-01-01
The article describes the experimental research of cement stone. Cement stone forming involves highly dispersive mineral additive - ground ash. It is stated that the substitution of some part of cement with activated ash leaves cement strength high. This is possible due to the activity of ash in structure forming processes. Activation of ash provides the increase in its puzzolanic activity, complete hydration processes. it is stated that ash grinding leads to a selective crystallization hydrated neoformations. Their morthology is different on outer and inner surfaces of ash spheres. The usage of ash can provide cement economy on condition that rheological characteristics of concrete stay constant. Besides, the usage of ash will improve physical and mechanic characteristics of cement stone and concrete.
78 FR 118 - Agency Information Collection Activities: Submission for OMB Review; Comment Request
Federal Register 2010, 2011, 2012, 2013, 2014
2013-01-02
... form number: NA Forms 13022, 13064, 13068. Type of review: Regular. Affected public: Former Federal... nature of the request. The NA Form 13022, Returned Request Form, is used to request additional information about the former Federal employee. The NA Form 13064, Reply to Request Involving Relief Agencies...
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2013-10-31
... Safety Testing of Sunlamp Products'' Form FDA 3632 ``Guide for Preparing Product Reports on Lasers and Products Containing Lasers'' Form FDA 3633''General Variance Request'' Form FDA 3634 ``Television Products Annual Report'' Form FDA 3635 ``Laser Light Show Notification'' Form FDA 3636 ``Guide for Preparing...
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2013-06-12
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2010-05-13
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75 FR 10025 - Proposed Collection; Comment Request for Forms 8038, 8038-G, and 8038-GC
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2010-03-04
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78 FR 1220 - Agency Information Collection Activities: Guarantee of Payment
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2013-01-08
... and approval in accordance with the Paperwork Reduction Act: Guarantee of Payment (CBP Form I-510... forms of information. Title: Guarantee of Payment. OMB Number: 1651-0127. Form Number: CBP Form I-510... master of the vessel or aircraft on CBP Form I-510, Guarantee of Payment. No vessel or aircraft can be...
Federal Register 2010, 2011, 2012, 2013, 2014
2011-07-20
... and avoid confusion for applicants. The currently approved forms are available online at http://www.../Upland Game Hunt Application). FWS Form 3-2357 (Migratory Bird Hunt Application). FWS Form 3-2358... seasons, as determined by State or Federal regulations. FWS Form 3-2359 (Big Game Harvest Report). FWS...
Sakai, T; Kawakatsu, H; Fujita, M; Yano, J; Owada, M K
1998-02-01
In previous work, we established a new monoclonal antibody that specifically recognizes the active form of c-Src tyrosine kinase (Kawakatsu et al, 1996). To determine whether c-Src is active in colorectal tumorigenesis, we examined the expression of an active form of c-Src in human normal mucosa, hyperplastic polyps, adenomas, and adenocarcinomas. The tissue distribution of the active form of c-Src was studied by immunohistochemistry using this antibody, termed Clone 28. Among 66 cases of adenoma tested, 61 (92%) showed positive staining (adenoma with mild atypia, 3 of 3; adenoma with moderate atypia, 38 of 42; adenoma with severe atypia, 20 of 21). In contrast to the frequent and intense staining in adenomas, adenocarcinoma showed weak staining with less frequency in 4 of 16 (25%) cases. The number of specimens with positive staining in well- and moderately differentiated adenocarcinomas was limited to an early stage. The active form of c-Src mainly localized to the nuclear membrane and the perinuclear region. These results provide the first direct evidence that the activation of c-Src appears to be an early event in colonic carcinogenesis in situ. The findings of the present study thus allow us to propose a molecular mechanism involving c-Src activation in the process of malignant transformation of the human colonic neoplastic cells.
ERIC Educational Resources Information Center
Reinke, Karen; Fernandes, Myra; Schwindt, Graeme; O'Craven, Kathleen; Grady, Cheryl L.
2008-01-01
The functional specificity of the brain region known as the Visual Word Form Area (VWFA) was examined using fMRI. We explored whether this area serves a general role in processing symbolic stimuli, rather than being selective for the processing of words. Brain activity was measured during a visual 1-back task to English words, meaningful symbols…
Legaz, Sophie; Exposito, Jean-Yves; Borel, Agnès; Candusso, Marie-Pierre; Megy, Simon; Montserret, Roland; Lahaye, Vincent; Terzian, Christophe; Verrier, Bernard
2015-09-01
Gal4/UAS system is a powerful tool for the analysis of numerous biological processes. Gal4 is a large yeast transcription factor that activates genes including UAS sequences in their promoter. Here, we have synthesized a minimal form of Gal4 DNA sequence coding for the binding and dimerization regions, but also part of the transcriptional activation domain. This truncated Gal4 protein was expressed as inclusion bodies in Escherichia coli. A structured and active form of this recombinant protein was purified and used to cover poly(lactic acid) (PLA) nanoparticles. In cellulo, these Gal4-vehicles were able to activate the expression of a Green Fluorescent Protein (GFP) gene under the control of UAS sequences, demonstrating that the decorated Gal4 variant can be delivery into cells where it still retains its transcription factor capacities. Thus, we have produced in E. coli and purified a short active form of Gal4 that retains its functions at the surface of PLA-nanoparticles in cellular assay. These decorated Gal4-nanoparticles will be useful to decipher their tissue distribution and their potential after ingestion or injection in UAS-GFP recombinant animal models. Copyright © 2015 Elsevier Inc. All rights reserved.
Effect of the chelation of metal cation on the antioxidant activity of chondroitin sulfates.
Ajisaka, Katsumi; Oyanagi, Yutaka; Miyazaki, Tatsuo; Suzuki, Yasuhiro
2016-06-01
The antioxidant potencies of chondroitin sulfates (CSs) from shark cartilage, salmon cartilage, bovine trachea, and porcine intestinal mucosa were compared by three representative methods for the measurement of the antioxidant activity; DPPH radical scavenging activity, superoxide radical scavenging activity, and hydroxyl radical scavenging activity. CSs from salmon cartilage and bovine trachea showed higher potency in comparison with CSs from shark cartilage and porcine intestinal mucosa. Next, CS from salmon cartilage chelating with Ca(2+), Mg(2+), Mn(2+), or Zn(2+) were prepared, and their antioxidant potencies were compared. CS chelating with Ca(2+) or Mg(2+) ions showed rather decreased DPPH radical scavenging activity in comparison with CS of H(+) form. In contrast, CS chelating with Ca(2+) or Mg(2+) ion showed remarkably enhanced superoxide radical scavenging activity than CS of H(+) or Na(+) form. Moreover, CS chelating with divalent metal ions, Ca(2+), Mg(2+), Mn(2+), or Zn(2+), showed noticeably higher hydroxyl radical scavenging activity than CS of H(+) or Na(+) form. The present results revealed that the scavenging activities of, at least, superoxide radical and hydroxyl radical were enhanced by the chelation with divalent metal ions.
Fujimaki, N; Miyauchi, S; Pütz, B; Sasaki, Y; Takino, R; Sakai, K; Tamada, T
1999-01-01
Functional magnetic resonance imaging was used to investigate neural activity during the judgment of visual stimuli in two groups of experiments using seven and five normal subjects. The subjects were given tasks designed differentially to involve orthographic (more generally, visual form), phonological, and lexico-semantic processes. These tasks included the judgments of whether a line was horizontal, whether a pseudocharacter or pseudocharacter string included a horizontal line, whether a Japanese katakana (phonogram) character or character string included a certain vowel, or whether a character string was meaningful (noun or verb) or meaningless. Neural activity related to the visual form process was commonly observed during judgments of both single real-characters and single pseudocharacters in lateral extrastriate visual cortex, the posterior ventral or medial occipito-temporal area, and the posterior inferior temporal area of both hemispheres. In contrast, left-lateralized activation was observed in the latter two areas during judgments of real- and pseudo-character strings. These results show that there is no katakana "word form center" whose activity is specific to real words. Activation related to the phonological process was observed, in Broca's area, the insula, the supramarginal gyrus, and the posterior superior temporal area, with greater activation in the left hemisphere. These activation foci for visual form and phonological processes of katakana also were reported for the English alphabet in previous studies. The present activation showed no additional areas for contrasts of noun judgment with other conditions and was similar between noun and verb judgment tasks, suggesting two possibilities: no strong semantic activation was produced, or the semantic process shared activation foci with the phonological process.
76 FR 71433 - Proposed Agency Information Collection Activities; Comment Request
Federal Register 2010, 2011, 2012, 2013, 2014
2011-11-17
... per Total annual Form Respondent universe responses response burden hours Form FRA F 6180.117 100 County (Regional) 2,750 forms...... 4 minutes........ 183 hours. Medical Examiners. Respondent Universe...
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75 FR 24959 - Proposed Information Collection Activity: Comment Request
Federal Register 2010, 2011, 2012, 2013, 2014
2010-05-06
... create or save millions of jobs. The ACF-196-477 TANF ARRA Financial Reporting form will Collect... Information Collection Activity: Comment Request Proposed Projects: Title: TANF ARRA Tribal Financial Reporting ACF-196T-477 form under Public Law 102-477 OMB No.: New Collection. Description: The ACF-196T-477...
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... Collection Activities; Proposed Collection; Comments Requested: Reporting and Recordkeeping for Digital... for digital certificates. (3) Agency form number, if any, and the applicable component of the.... Abstract: Persons use these forms to apply for DEA-issued digital certificates to order Schedule I and II...
Compulsory "Foreign Language Activities" in Japanese Primary Schools
ERIC Educational Resources Information Center
Hashimoto, Kayoko
2011-01-01
From 2011, the new curriculum for introducing English to Japanese primary schools will be fully implemented in the form of "foreign language activities". This innovation forms part of the government's plan to cultivate "Japanese with English abilities", a development based on the awareness, particularly in the business sector,…
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2011-03-18
... (Certification of School Attendance or Termination) Activity: Comment Request AGENCY: Veterans Benefits... through the use of automated collection techniques or the use of other forms of information technology. Title: Certification of School Attendance or Termination, VA Forms 21-8960 and 21-8960-1. OMB Control...
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2013-04-26
... OFFICE OF GOVERNMENT ETHICS Agency Information Collection Activities; Submission for OMB Review; Proposed Collection; Comment Request for a Modified OGE Form 201 Ethics in Government Act Access Form AGENCY: Office of Government Ethics (OGE). ACTION: Notice of request for agency and public comments...
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2013-09-13
... OFFICE OF GOVERNMENT ETHICS Agency Information Collection Activities; Submission for OMB Review; Proposed Collection; Comment Request for a Modified OGE Form 201 Ethics in Government Act Access Form AGENCY: Office of Government Ethics (OGE). ACTION: Notice. SUMMARY: After publication of this second...
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2010-03-02
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78 FR 52235 - Proposed Collection; Comment Request for Form 8825
Federal Register 2010, 2011, 2012, 2013, 2014
2013-08-22
... Form 8825, Rental Real Estate Income and Expenses of a Partnership or an S Corporation. DATES: [email protected] . SUPPLEMENTARY INFORMATION: Title: Rental Real Estate Income and Expenses of a... rental real estate activities, including net income or loss from rental real estate activities that flow...
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2013-04-12
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2013-12-24
...-0018] Agency Information Collection Activities: Application for Permission To Reapply for Admission... Collection. (2) Title of the Form/Collection: Application for Permission to Reapply for Admission into the... on Form I-212 is used by USCIS to adjudicate applications filed by aliens requesting consent to...
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2012-07-24
...-0018] Agency Information Collection Activities: Application for Permission To Reapply for Admission... the Form/Collection: Application for Permission To Reapply for Admission Into the United States After...: Individuals or households. USCIS uses the information provided on Form I-212 to adjudicate applications filed...
Cultural Dance: An Opportunity To Encourage Physical Activity and Health in Communities.
ERIC Educational Resources Information Center
Jain, Sheila; Brown, David R.
2001-01-01
Evaluated the literature related to dance forms having sociocultural or traditional/ceremonial influences (cultural dance). Research documented the benefits of dance- related cultural activity interventions among various subgroups of the population and the role and importance of cultural dance forms within diverse communities. The scant available…
78 FR 75576 - Agency Information Collection Activities: Importation Bond Structure
Federal Register 2010, 2011, 2012, 2013, 2014
2013-12-12
... Activities: Importation Bond Structure AGENCY: U.S. Customs and Border Protection (CBP), Department of... requirement concerning the Importation Bond Structure. This request for comment is being made pursuant to the...: Title: Importation Bond Structure. OMB Number: 1651-0050. Form Number: CBP Forms 301 and 5297. Abstract...
77 FR 11494 - Proposed Collection; Comment Request
Federal Register 2010, 2011, 2012, 2013, 2014
2012-02-27
... Defense Pentagon, Washington, DC 20301-4000 or call (703) 588-0877. Title, Associated Form, and OMB... Certified to Have Performed Active Duty with the Armed Forces of the United States, DD Form 2168, OMB... the United States by certain groups shall be considered Active Duty service, and (2) to award members...
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2012-10-02
...: 2120-0018. Title: Certification Procedures for Products and Parts. Form Numbers: FAA Forms 8110-12... Activities: Requests for Comments; Clearance of Renewed Approval of Information Collection: Certification Procedures for Products and Parts AGENCY: Federal Aviation Administration (FAA), DOT. ACTION: Notice and...
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2013-01-28
... Title: Certification Procedures for Products and Parts Form Numbers: FAA Forms 8110-12, 8130-1, 8130-6... Activities: Requests for Comments; Clearance of Renewed Approval of Information Collection: Certification Procedures for Products and Parts AGENCY: Federal Aviation Administration (FAA), DOT. ACTION: Notice and...
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2010-09-27
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76 FR 72934 - Proposed Information Collection Activity; Comment Request
Federal Register 2010, 2011, 2012, 2013, 2014
2011-11-28
... Information Collection Activity; Comment Request Title: ACF-OGM-SF-PPR-Form B--Program Indicators. OMB No. New Collection. Description The Office of Grants Management (OGM), in the Administration for Children and... collect this data OGM has developed a form from the basic template of the OMB-approved reporting format of...
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2011-02-08
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76 FR 60134 - Agency Information Collection (Child Care Subsidy) Activity Under OMB Review
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2011-09-28
... Care Subsidy) Activity Under OMB Review AGENCY: Human Resources and Administration, Department of....gov . Please refer to ``OMB Control No. 2900-0717.'' SUPPLEMENTARY INFORMATION: Titles a. Child Care Subsidy Application Form, VA Form 0730a. b. Child Care Provider Information (For the Child Care Subsidy...
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2013-08-26
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2013-07-31
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2010-10-07
... (Application for Reimbursement of National Test Fee) Activity: Comment Request AGENCY: Veterans Benefits.... Title: Application for Reimbursement of National Test Fee, VA Form 22-0810. OMB Control Number: 2900..., and eligible dependents complete VA Form 22-0810 to request reimbursement of national test fees. VA...
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2010-07-06
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2012-11-30
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2011-10-05
... DEPARTMENT OF HEALTH AND HUMAN SERVICES Food and Drug Administration [Docket No. FDA-2011-N-0708] Agency Information Collection Activities; Proposed Collection; Comment Request; FDA Form 3728, Animal...: The Food and Drug Administration (FDA) is announcing an opportunity for public comment on the proposed...
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2013-03-19
... Collection Activities: Comment Request for the Mine, Development, and Mineral Exploration Supplement (1 Form..., Development, and Mineral Exploration Supplement. This collection consists of one form and this notice provides... with domestic production, exploration, and mine development data for nonfuel mineral commodities. This...
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2013-12-03
... Information Collection Activities: Reinstatement, Without Change; Comment Request ACTION: 30-Day Notice of Information Collection for Review; Form No. I- 333, Obligor Change of Address; OMB Control No. 1653-0042. The... appropriate automated, electronic, mechanical, or other technological collection techniques or other forms of...
75 FR 9022 - Submission for OMB Review; Comment Request
Federal Register 2010, 2011, 2012, 2013, 2014
2010-02-26
... Review: Extension of a currently approved collection. Title: Passive Activity Credit Limitations. Form: 8582-CR. Description: Under section 469, credits from passive activities, to the extent they do not exceed the tax attributable to net passive income, are not allowed. Form 8582-CR is used to figure the...
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2011-07-12
...] Agency Information Collection Activities: Proposed Collection Comments Requested Race and National Origin... collection. (2) Title of the Form/Collection: Race and National Origin Identification. (3) Agency form number.... Other: none. Need for Collection: The information collection is used to maintain Race and National...
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2011-06-21
... INFORMATION: OMB Control Number: 2120-0042. Title: Aircraft Registration. Form Numbers: FAA Forms 8050-1, 8050.../or revocation of registration. The information collected is used by the FAA to register an aircraft... Activities: Requests for Comments; Clearance of Renewed Approval of Information Collection: Aircraft...
C-H bond activation of hydrocarbons by an imidozirconocene complex.
Hoyt, Helen M; Michael, Forrest E; Bergman, Robert G
2004-02-04
Monomeric imidozirconocene complexes of the type Cp2(L)Zr=NCMe3 (Cp = cyclopentadienyl, L = Lewis base) have been shown to activate the carbon-hydrogen bonds of benzene, but not the C-H bonds of saturated hydrocarbons. To our knowledge, this singularly important class of C-H activation reactions has heretofore not been observed in imidometallocene systems. The M=NR bond formed on heating the racemic ethylenebis(tetrahydro)indenyl methyl tert-butyl amide complex, however, cleanly and quantitatively activates a wide range of n-alkane, alkene, and arene C-H bonds. Mechanistic experiments support the proposal of intramolecular elimination of methane followed by a concerted addition of the hydrocarbon C-H bond. Products formed by activation of sp2 C-H bonds are generally more thermodynamically stable than those formed by activation of sp3 C-H bonds, and those resulting from reaction at primary C-H bonds are preferred over secondary sp3 C-H activation products. There is also evidence that thermodynamic selectivity among C-H bonds is sterically rather than electronically controlled.
Comparative analysis of activator-Eσ54 complexes formed with nucleotide-metal fluoride analogues
Burrows, Patricia C.; Joly, Nicolas; Nixon, B. Tracy; Buck, Martin
2009-01-01
Bacterial RNA polymerase (RNAP) containing the major variant σ54 factor forms open promoter complexes in a reaction in which specialized activator proteins hydrolyse ATP. Here we probe binding interactions between σ54-RNAP (Eσ54) and the ATPases associated with various cellular activities (AAA+) domain of the Escherichia coli activator protein, PspF, using nucleotide-metal fluoride (BeF and AlF) analogues representing ground and transition states of ATP, which allow complexes (that are otherwise too transient with ATP) to be captured. We show that the organization and functionality of the ADP–BeF- and ADP–AlF-dependent complexes greatly overlap. Our data support an activation pathway in which the initial ATP-dependent binding of the activator to the Eσ54 closed complex results in the re-organization of Eσ54 with respect to the transcription start-site. However, the nucleotide-dependent binding interactions between the activator and the Eσ54 closed complex are in themselves insufficient for forming open promoter complexes when linear double-stranded DNA is present in the initial closed complex. PMID:19553192
Large-scale filament formation inhibits the activity of CTP synthetase
Barry, Rachael M; Bitbol, Anne-Florence; Lorestani, Alexander; Charles, Emeric J; Habrian, Chris H; Hansen, Jesse M; Li, Hsin-Jung; Baldwin, Enoch P; Wingreen, Ned S; Kollman, Justin M; Gitai, Zemer
2014-01-01
CTP Synthetase (CtpS) is a universally conserved and essential metabolic enzyme. While many enzymes form small oligomers, CtpS forms large-scale filamentous structures of unknown function in prokaryotes and eukaryotes. By simultaneously monitoring CtpS polymerization and enzymatic activity, we show that polymerization inhibits activity, and CtpS's product, CTP, induces assembly. To understand how assembly inhibits activity, we used electron microscopy to define the structure of CtpS polymers. This structure suggests that polymerization sterically hinders a conformational change necessary for CtpS activity. Structure-guided mutagenesis and mathematical modeling further indicate that coupling activity to polymerization promotes cooperative catalytic regulation. This previously uncharacterized regulatory mechanism is important for cellular function since a mutant that disrupts CtpS polymerization disrupts E. coli growth and metabolic regulation without reducing CTP levels. We propose that regulation by large-scale polymerization enables ultrasensitive control of enzymatic activity while storing an enzyme subpopulation in a conformationally restricted form that is readily activatable. DOI: http://dx.doi.org/10.7554/eLife.03638.001 PMID:25030911
Processing of metacaspase 2 from Trypanosoma brucei (TbMCA2) broadens its substrate specificity.
Gilio, Joyce M; Marcondes, Marcelo F; Ferrari, Débora; Juliano, Maria A; Juliano, Luiz; Oliveira, Vitor; Machado, Maurício F M
2017-04-01
Metacaspases are members of the cysteine peptidase family and may be implicated in programmed cell death in plants and lower eukaryotes. These proteases exhibit calcium-dependent activity and specificity for arginine residues at P 1 . In contrast to caspases, they do not require processing or dimerization for activity. Indeed, unprocessed metacaspase-2 of Trypanosoma brucei (TbMCA2) is active; however, it has been shown that cleavages at Lys 55 and Lys 268 increase TbMCA2 hydrolytic activity on synthetic substrates. The processed TbMCA2 comprises 3 polypeptide chains that remain attached by non-covalent bonds. Replacement of Lys 55 and Lys 268 with Gly via site-directed mutagenesis results in non-processed but enzymatically active mutant, TbMCA2 K55/268G. To investigate the importance of this processing for the activity and specificity of TbMCA2, we performed activity assays comparing the non-processed mutant (TbMCA2 K55/268G) with the processed TbMCA2 form. Significant differences between TbMCA2 WT (processed form) and TbMCA2 K55/268G (non-processed form) were observed. Specifically, we verified that although non-processed TbMCA2 is active when assayed with small synthetic substrates, the TbMCA2 form does not exhibit hydrolytic activity on large substrates such as azocasein, while processed TbMCA2 is able to readily digest this protein. Such differences can be relevant for understanding the physiological regulation and function of TbMCA2. Copyright © 2017 Elsevier B.V. All rights reserved.
Spolarics, Z; Bond, J S
1989-11-01
Arginase activity is elevated in livers of diabetic animals compared to controls and there is evidence that this is due in part to increased specific activity (activity/mg arginase protein). To investigate the molecular basis of this increased activity, the physicochemical and kinetic properties of hepatic arginase from diabetic and control mice were compared. Two types of arginase subunits with molecular weights of 35,000 and 38,000 were found in both the diabetic and control animals and the subunits in these animals had similar, multiple ionic forms. Kinetic parameters of purified preparations of arginase for arginine (apparent Km and Vmax values) and the thermal stability of these preparations from diabetics and controls were also similar. Furthermore, no difference was found in the distribution of arginase activity among different subcellular liver fractions. Separation of basic and acidic oligomeric forms of arginase by fast-protein liquid chromatography resulted in a slightly different distribution of activity among the forms in the normal and diabetic group. The apparent Km values for Mn2+ of the basic form of the enzyme were 25 and 33 microM for the enzyme from normal and diabetic animals, respectively; for acidic forms, for which two apparent Km values were measured, the values were 8 and 197 microM for arginase from controls and 35 and 537 microM from diabetics. These results indicate that in diabetes, while no marked changes in the physicochemical characteristics of arginase are obvious, some changes are found in the interaction of arginase with its cofactor Mn.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Aoyama, Toshifumi; Yamano, Shigeru; Gelboin, H.V.
Twelve forms of human hepatic cytochrome P450 were expressed in hepatoma cells by means of recombinant vaccinia viruses. The expressed P450s were analyzed for their abilities to activate the potent hepatocarcinogen aflatoxin B{sub 1} to metabolites having mutagenic or DNA-binding properties. Five forms, P450s IA2, IIA3, IIB7, IIIA3, and IIIA4, activated aflatoxin B{sub 1} to mutagenic metabolites as assessed by the production of His revertants of Salmonella typhimurium in the Ames test. The same P450s catalyzed conversion of aflatoxin B{sub 1} to DNA-bound derivatives as judged by an in situ assay in which the radiolabeled carcinogen was incubated with cellsmore » expressing the individual P450 forms. Seven other human P450s, IIC8, IIC9, IID6, IIE1, IIF1, and IIIA5, and IVB1, did not significantly activate aflatoxin B{sub 1} as measured by both the Ames test and the DNA-binding assay. Moreover, polyclonal anti-rat liver P450 antibodies that crossreact with individual human P450s IA2, IIA3, IIIA3, and IIIA4 each inhibited aflatoxin B{sub 1} activation catalyzed by human liver S-9 extracts. Inhibition ranged from as low as 10% with antibody against IIA3 to as high as 65% with antibody against IIIA3 and IIIA4. These results establish that metabolic activation of aflatoxin B{sub 1} in human liver involves the contribution of multiple forms of P450.« less
Dixit, Anshuman; Verkhivker, Gennady M.
2009-01-01
Structural and functional studies of the ABL and EGFR kinase domains have recently suggested a common mechanism of activation by cancer-causing mutations. However, dynamics and mechanistic aspects of kinase activation by cancer mutations that stimulate conformational transitions and thermodynamic stabilization of the constitutively active kinase form remain elusive. We present a large-scale computational investigation of activation mechanisms in the ABL and EGFR kinase domains by a panel of clinically important cancer mutants ABL-T315I, ABL-L387M, EGFR-T790M, and EGFR-L858R. We have also simulated the activating effect of the gatekeeper mutation on conformational dynamics and allosteric interactions in functional states of the ABL-SH2-SH3 regulatory complexes. A comprehensive analysis was conducted using a hierarchy of computational approaches that included homology modeling, molecular dynamics simulations, protein stability analysis, targeted molecular dynamics, and molecular docking. Collectively, the results of this study have revealed thermodynamic and mechanistic catalysts of kinase activation by major cancer-causing mutations in the ABL and EGFR kinase domains. By using multiple crystallographic states of ABL and EGFR, computer simulations have allowed one to map dynamics of conformational fluctuations and transitions in the normal (wild-type) and oncogenic kinase forms. A proposed multi-stage mechanistic model of activation involves a series of cooperative transitions between different conformational states, including assembly of the hydrophobic spine, the formation of the Src-like intermediate structure, and a cooperative breakage and formation of characteristic salt bridges, which signify transition to the active kinase form. We suggest that molecular mechanisms of activation by cancer mutations could mimic the activation process of the normal kinase, yet exploiting conserved structural catalysts to accelerate a conformational transition and the enhanced stabilization of the active kinase form. The results of this study reconcile current experimental data with insights from theoretical approaches, pointing to general mechanistic aspects of activating transitions in protein kinases. PMID:19714203
78 FR 39832 - Proposed Information Collection Activity: Proposed Collection; Comment Request
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... techniques or the use of other forms of information technology. Titles: a. Application for Health Benefits, VA Form 10-10EZ. b. Health Benefits Renewal Form, VA Form 10-10EZR. c. VA Form 10-10HS. OMB Control... determine the Veteran's eligibility for medical benefits. b. Veterans currently enrolled in VA health care...
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Tajeddin, Zia; Daraee, Dina
2013-01-01
The present study investigated the effect of form-focused and non-form-focused tasks on EFL learners' vocabulary learning through written input. The form-focused task aimed to draw students' attention to the word itself through word recognition activities. Non-form-focused tasks were divided into (a) the comprehension question task, which required…
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The purpose of this SOP is to define the steps for creating a scannable form and defining the database associated with the activated form. It applies to forms developed for the Arizona NHEXAS project and the "Border" study. Keywords: data; scannable forms.
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NASA Astrophysics Data System (ADS)
Gryzunova, N. N.; Vikarchuk, A. A.; Gryzunov, A. M.; Denisova, A. G.
2017-10-01
The morphology of the electrolytic copper single crystals formed under the mechanical activation of a cathode is described. Pentagonal pyramids and conical microcrystals with high growth steps are shown to form during electrocrystallization under these conditions. It is experimentally found that microcrystals grow on disclination defects, in particular, at the sites of termination of twin growth boundaries, and mechanical activation causes the formation of such defects.
Inhibitory effect of chalcone derivatives on recombinant human aldose reductase.
Iwata, S; Nagata, N; Omae, A; Yamaguchi, S; Okada, Y; Shibata, S; Okuyama, T
1999-03-01
More than fifty chalcone derivatives were synthesized to examine structure-activity relationships against human aldose reductase. Certain 2',4'-dihydroxychalcone derivatives inhibited human aldose reductase activities, and 2',4',2, 4-tetrahydroxychalcone and 2',4',2-trihydroxychalcone showed potent inhibitory activity with IC50 values of 7.4x10(-9) M and 1.6x10(-7) M, respectively. On the other hand, cis-form chalcones, which were isomerized from the original trans-forms by irradiation of daylight in methanol solution, promoted the activity of human aldose reductase.
Trypanocidal activity of extracts from Brazilian Atlantic Rain Forest plant species.
Pizzolatti, M G; Koga, A H; Grisard, E C; Steindel, M
2003-01-01
The trypanocidal activity of crude hydro alcoholic extracts and several fractions of 13 plants from Brazilian Atlantic Rain Forest were tested in vitro against epimastigote and trypomastigote forms of Trypanosoma cruzi, the etiological agent of Chagas disease. Crude ethanol extracts with promising in vitro activity (DL50 between 5-10 microg/ml) against epimastigotes were fractionated by solvent partition and further tested against bloodstream form of the parasite. Activity against bloodstream parasites was observed in both dichloromethane and hexane fractions of Polygala sabulosa and P. paniculata.
Formation of alcohol conversion catalysts
Wachs, Israel E.; Cai, Yeping
2001-01-01
The method of the present invention involves a composition containing an intimate mixture of (a) metal oxide support particles and (b) a catalytically active metal oxide from Groups VA, VIA, or VIIA, its method of manufacture, and its method of use for converting alcohols to aldehydes. During the conversion process, catalytically active metal oxide from the discrete catalytic metal oxide particles migrates to the oxide support particles and forms a monolayer of catalytically active metal oxide on the oxide support particle to form a catalyst composition having a higher specific activity than the admixed particle composition.
Method for making electro-fluidic connections in microfluidic devices
Frye-Mason, Gregory C.; Martinez, David; Manginell, Ronald P.; Heller, Edwin J.; Chanchani, Rajen
2004-08-10
A method for forming electro-fluidic interconnections in microfluidic devices comprises forming an electrical connection between matching bond pads on a die containing an active electrical element and a microfluidic substrate and forming a fluidic seal ring that circumscribes the active electrical element and a fluidic feedthrough. Preferably, the electrical connection and the seal ring are formed in a single bonding step. The simple method is particularly useful for chemical microanalytical systems wherein a plurality of microanalytical components, such as a chemical preconcentrator, a gas chromatography column, and a surface acoustic wave detector, are fluidically interconnected on a hybrid microfluidic substrate having electrical connection to external support electronics.
Marine Omega-3 Phospholipids: Metabolism and Biological Activities
Burri, Lena; Hoem, Nils; Banni, Sebastiano; Berge, Kjetil
2012-01-01
The biological activities of omega-3 fatty acids (n-3 FAs) have been under extensive study for several decades. However, not much attention has been paid to differences of dietary forms, such as triglycerides (TGs) versus ethyl esters or phospholipids (PLs). New innovative marine raw materials, like krill and fish by-products, present n-3 FAs mainly in the PL form. With their increasing availability, new evidence has emerged on n-3 PL biological activities and differences to n-3 TGs. In this review, we describe the recently discovered nutritional properties of n-3 PLs on different parameters of metabolic syndrome and highlight their different metabolic bioavailability in comparison to other dietary forms of n-3 FAs. PMID:23203133
Efficient renaturation of inclusion body proteins denatured by SDS.
He, Chuan; Ohnishi, Kouhei
2017-09-02
Inclusion bodies are often formed when the foreign protein is over expressed in Escherichia coli. Since proteins in inclusion bodies are inactive, denaturing and refolding of inclusion body proteins are necessary to obtain the active form. Instead of the conventional denaturants, urea and guanidine hydrochloride, a strong anionic detergent SDS was used to solubilize C-terminal His-tag form of ulvan lyase in the inclusion bodies. Solution containing SDS-solubilized enzyme were kept on ice to precipitate SDS, followed by SDS-KCl insoluble crystal formation to remove SDS completely. After removing the precipitate by centrifugation, the supernatant was applied to Ni-NTA column to purify His-tagged ulvan lyase. The purified protein showed a dimeric form and ulvan lyase activity, demonstrating that SDS-denatured protein was renatured and recovered enzyme activity. This simple method could be useful for refolding other inclusion body proteins. Copyright © 2017 Elsevier Inc. All rights reserved.
Cloud, Veronica; Chan, Yuen-Ling; Grubb, Jennifer; Budke, Brian; Bishop, Douglas K.
2014-01-01
Meiotic recombination in budding yeast requires two RecA-related proteins, Rad51 and Dmc1, both of which form filaments on DNA capable of directing homology search and catalyzing formation of homologous joint molecules (JMs) and strand exchange. Using a separation-of-function mutant form of Rad51, that retains filament-forming but not JM forming activity, we show that the JM activity of Rad51 is fully dispensable for meiotic recombination. The corresponding mutation in Dmc1 causes a profound recombination defect, demonstrating Dmc1’s JM activity alone is responsible for meiotic recombination. We further provide biochemical evidence that Rad51 acts with Mei5-Sae3 as a Dmc1 accessory factor. Thus, Rad51 is a multifunctional protein that catalyzes recombination directly in mitosis and indirectly, via Dmc1, during meiosis. PMID:22955832
Rad51 is an accessory factor for Dmc1-mediated joint molecule formation during meiosis.
Cloud, Veronica; Chan, Yuen-Ling; Grubb, Jennifer; Budke, Brian; Bishop, Douglas K
2012-09-07
Meiotic recombination in budding yeast requires two RecA-related proteins, Rad51 and Dmc1, both of which form filaments on DNA capable of directing homology search and catalyzing formation of homologous joint molecules (JMs) and strand exchange. With use of a separation-of-function mutant form of Rad51 that retains filament-forming but not JM-forming activity, we show that the JM activity of Rad51 is fully dispensable for meiotic recombination. The corresponding mutation in Dmc1 causes a profound recombination defect, demonstrating Dmc1's JM activity alone is responsible for meiotic recombination. We further provide biochemical evidence that Rad51 acts with Mei5-Sae3 as a Dmc1 accessory factor. Thus, Rad51 is a multifunctional protein that catalyzes recombination directly in mitosis and indirectly, via Dmc1, during meiosis.
Interfacial activity in alkaline flooding enhanced oil recovery
DOE Office of Scientific and Technical Information (OSTI.GOV)
Chan, M.K.
1981-01-01
The ionization of long-chained organic acids in the crude oil to form soaps was shown to be primarily responsible for the lowering of oil-water interfacial tension at alkaline pH. These active acids can be concentrated by silica gel chromatography into a minor polar fraction. An equilibrium chemical model was proposed based on 2 competing reactions: the ionization of acids to form active anions, and the formation of undissociated soap between acid anions and sodium ions. It correlates the interfacial activity with the interfacial concentration of active acid anions which is expressed in terms of the concentrations of the chemical speciesmore » in the system. The model successfully predicts the observed oil-alkaline solution interfacial phenomenon, including its dependence on pH, alkali and salt concentrations, type of acid present and type of soap formed. Flooding at different alkali concentrations to activate different acid species present in the crude was shown to give better recovery than flooding at a single high alkali concentration. Treating the crude oil with a dilute solution of mineral acids liberates additional free active acids and yields better interfacial activity during subsequent alkali contact.« less
Typology of social space in Kauman Kampong Semarang
NASA Astrophysics Data System (ADS)
Endrianto Pandelaki, Edward; Suprapti, Atiek; Wahyu Firmandhani, Satriya
2017-12-01
Social space is various forms of spaces used by community in conducting social interaction. These kinds of spaces should be given more attention since they serve as catalyst in the implementation of good social cohesion in community. The effort includes giving concern toward their various implemented form. This concern will give benefits in the creation of resilient built environment. Kauman Kampong in Semarang, is an old urban kampong and still exist up until now. During its development, the inhabitant live and conduct their activities in good shape. Therefore this kampong is an appropriate place to learn and explore social spaces which is formed and utilized by the community who conduct their activities in this kampong. The aim of this research is to find out forms and typology of social space in Kauman Kampong in Semarang. Qualitative method is used in this research since the nature of this research is explorative. There are various social activities in Kauman Kampong in Semarang, such as religious, trading, and other social interaction, which have formed various social spaces. These social spaces have their own physical characteristics and with various intensity of activities. Based on collected data in field survey, the typology of social spaces that could be inferred are: permanent, temporary, and dynamic social space.
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Active Inference, Epistemic Value, and Vicarious Trial and Error
ERIC Educational Resources Information Center
Pezzulo, Giovanni; Cartoni, Emilio; Rigoli, Francesco; io-Lopez, Léo; Friston, Karl
2016-01-01
Balancing habitual and deliberate forms of choice entails a comparison of their respective merits--the former being faster but inflexible, and the latter slower but more versatile. Here, we show that arbitration between these two forms of control can be derived from first principles within an Active Inference scheme. We illustrate our arguments…
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Process for forming sulfuric acid
Lu, Wen-Tong P.
1981-01-01
An improved electrode is disclosed for the anode in a sulfur cycle hydrogen generation process where sulfur dioxie is oxidized to form sulfuric acid at the anode. The active compound in the electrode is palladium, palladium oxide, an alloy of palladium, or a mixture thereof. The active compound may be deposited on a porous, stable, conductive substrate.
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Expressing our internal states and understanding those of others
Di Cesare, Giuseppe; Di Dio, Cinzia; Marchi, Massimo; Rizzolatti, Giacomo
2015-01-01
Vitality form is a term that describes the style with which motor actions are performed (e.g., rude, gentle, etc.). They represent one characterizing element of conscious and unconscious bodily communication. Despite their importance in interpersonal behavior, vitality forms have been, until now, virtually neglected in neuroscience. Here, using the functional MRI (fMRI) technique, we investigated the neural correlates of vitality forms in three different tasks: action observation, imagination, and execution. Conjunction analysis showed that, in all three tasks, there is a common, consistent activation of the dorsocentral sector of the insula. In addition, a common activation of the parietofrontal network, typically active during arm movements production, planning, and observation, was also found. We conclude that the dorsocentral part of the insula is a key element of the system that modulates the cortical motor activity, allowing individuals to express their internal states through action vitality forms. Recent monkey anatomical data show that the dorsocentral sector of the insula is, indeed, connected with the cortical circuit involved in the control of arm movements. PMID:26243879
Amyloid-β oligomer Aβ*56 induces specific alterations of tau phosphorylation and neuronal signaling
Amar, Fatou; Sherman, Mathew A.; Rush, Travis; Larson, Megan; Boyle, Gabriel; Chang, Liu; Götz, Jürgen; Buisson, Alain; Lesné, Sylvain E.
2018-01-01
Oligomeric forms of amyloid-forming proteins are believed to be the principal initiating bioactive species in many neurodegenerative disorders, including Alzheimer’s disease (AD). Amyloid-β (Aβ) oligomers are implicated in pathological modification and aggregation of the microtubule-associated protein tau. To investigate the specific molecular pathways activated by different assemblies, we isolated various forms of Aβ from Tg2576 mice. We found that the Aβ*56, which is linked with preclinical AD, interacted with NMDA receptors (NMDARs) in primary cortical neurons, increased NMDAR-dependent Ca2+ influx and, consequently, increased intracellular calcium concentrations and the activation of Ca2+-dependent calmodulin kinase IIα (CaMKIIα). In neurons in mice and in culture, activated CaMKIIα induced increased phosphorylation and missorting of tau, which is associated with AD pathology. In contrast, exposure of cultured primary cortical neurons to other oligomeric Aβ forms (dimers and trimers) did not trigger these effects. Our results indicate that distinct Aβ assemblies activate neuronal signaling pathways in a selective manner, and that dissecting the molecular events caused by each may inform more effective therapeutic strategies. PMID:28487416
A novel proteolytic processing of prolysyl oxidase
Atsawasuwan, Phimon; Mochida, Yoshiyuki; Katafuchi, Michitsuna; Tokutomi, Kentaro; Mocanu, Viorel; Parker, Carol E.; Yamauchi, Mitsuo
2012-01-01
Lysyl oxidase (LOX) is an amine oxidase that is critical for the stability of connective tissues. The secreted proLOX is enzymatically quiescent and is activated through proteolytic cleavage between residue Gly162 and Asp163 (residue numbers according to the mouse LOX) by bone morphogenetic protein (BMP)-1 gene products. Here we report a novel processing of proLOX identified in vitro and in vivo. Two forms of mature LOX were identified and characterized by their immunoreactivity to specific antibodies, amine oxidase activity and mass spectrometry. One form was identified as a well characterized BMP-1 processed LOX protein. Another was found to be a truncated form of LOX (tLOX) resulting from the cleavage at the carboxy terminus of Arg192. The tLOX still appeared to retain amine oxidase activity. The results from the proLOX gene deletion and mutation experiments indicated that the processing occurs independent of the cleavage of proLOX by BMP-1 gene products and likely requires the presence of LOX propeptide. These results indicate that proLOX could be processed by two different mechanisms producing two forms of active LOX. PMID:21591931
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Quantal concept of T-cell activation: adhesion domains as immunological synapses
NASA Astrophysics Data System (ADS)
Sackmann, Erich
2011-06-01
Adhesion micro-domains (ADs) formed during encounters of lymphocytes with antigen-presenting cells (APC) mediate the genetic expression of quanta of cytokines interleukin-2 (IL-2). The IL-2-induced activation of IL-2 receptors promotes the stepwise progression of the T-cells through the cell cycle, hence their name, immunological synapses. The ADs form short-lived reaction centres controlling the recruitment of activators of the biochemical pathway (the kinases Lck and ZAP) while preventing the access of inhibitors (phosphatase CD45) through steric repulsion forces. CD45 acts as the generator of adhesion domains and, through its role as a spacer protein, also as the promoter of the reaction. In a second phase of T-cell-APC encounters, long-lived global reaction spaces (called supramolecular activation complexes (SMAC)) form by talin-mediated binding of the T-cell integrin (LFA-1) to the counter-receptor ICAM-1, resulting in the formation of ring-like tight adhesion zones (peripheral SMAC). The ADs move to the centre of the intercellular adhesion zone forming the central SMAC, which serve in the recycling of the AD. We propose that cell stimulation is triggered by integrating the effect evoked by the short-lived adhesion domains. Similar global reaction platforms are formed by killer cells to destruct APC. We present a testable mechanical model showing that global reaction spaces (SMAC or dome-like contacts between cytotoxic cells and APC) form by self-organization through delayed activation of the integrin-binding affinity and stabilization of the adhesion zones by F-actin recruitment. The mechanical stability and the polarization of the adhering T-cells are mediated by microtubule-actin cross-talk.
Heterosubunit composition and crystal structures of a novel bacterial M16B metallopeptidase.
Maruyama, Yukie; Chuma, Asako; Mikami, Bunzo; Hashimoto, Wataru; Murata, Kousaku
2011-03-18
Three subfamilies of metallopeptidase family M16 enzymes--M16A, M16B, and M16C--are widely distributed among eukaryotes and prokaryotes. SPH2681, a periplasmic M16B protein found in Sphingomonas sp. strain A1, contains an HXXEH motif essential for Zn(2+) binding and catalytic activity. SPH2682 is another member of M16B, which lacks the metal-binding motif but conserves an active-site R/Y pair commonly found in the C-terminal half of M16 enzymes. Two genes coding for SPH2681 and SPH2682 assemble into a single operon in the bacterial genome. This study determined SPH2681 to be constitutively expressed in strain A1 cells grown on different carbon sources, suggesting a more general cellular function. SPH2681 and SPH2681/SPH2682 were overexpressed in Escherichia coli, purified, and characterized. SPH2681 was found to associate with SPH2682, forming a heterosubunit enzyme with peptidase activity, while SPH2681 alone exhibited no enzymatic activity. X-ray crystallography of the SPH2681/SPH2682 complex revealed two conformations (open and closed heterodimeric forms) within the same crystal. Compared with the closed form, the open form contains two subunits rotated away from each other by approximately 8°, increasing the distance between the zinc ion and active-site residues by up to 8 Å. In addition, many hydrogen bonds are formed or broken on change between the conformations of the heterodimers, suggesting that subunit dynamics is a prerequisite for catalysis. To our knowledge, this is the first report on both conformational forms of the same M16 peptidase, providing a unique insight into the general proteolytic mechanism of M16 proteases. Copyright © 2011 Elsevier Ltd. All rights reserved.
Miltefosine is effective against Candida albicans and Fusarium oxysporum nail biofilms in vitro.
Machado Vila, Taissa Vieira; Sousa Quintanilha, Natália; Rozental, Sonia
2015-11-01
Onychomycosis is a fungal nail infection that represents ∼50 % of all nail disease cases worldwide. Clinical treatment with standard antifungals frequently requires long-term systemic therapy to avoid chronic disease. Onychomycosis caused by non-dermatophyte moulds, such as Fusarium spp., and yeasts, such as Candida spp., is particularly difficult to treat, possibly due to the formation of drug-resistant fungal biofilms on affected areas. Here, we show that the alkylphospholipid miltefosine, used clinically against leishmaniasis and cutaneous breast metastases, has potent activity against biofilms of Fusarium oxysporum and Candida albicans formed on human nail fragments in vitro. Miltefosine activity was compared with that of commercially available antifungals in the treatment of biofilms at two distinct developmental phases: formation and maturation (pre-formed biofilms). Drug activity towards biofilms formed on nail fragments and on microplate surfaces (microdilution assays) was evaluated using XTT [2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide] assays, and drug effects on fingernail biofilms were analysed by scanning electron microscopy (SEM). For F. oxysporum, miltefosine at 8 μg ml- 1 inhibited biofilm formation by 93%, whilst 256 μg ml- 1 reduced the metabolic activity of pre-formed nail biofilms by 93%. Treatment with miltefosine at 1000 μg ml- 1 inhibited biofilm formation by 89% and reduced the metabolic activity of pre-formed C. albicans biofilms by 99%. SEM analyses of biofilms formed on fingernail fragments showed a clear reduction in biofilm biomass after miltefosine treatment, in agreement with XTT results. Our results show that miltefosine has potential as a therapeutic agent against onychomycosis and should be considered for in vivo efficacy studies, especially in topical formulations for refractory disease treatment.
Jóźwik, Artur; Marchewka, Joanna; Strzałkowska, Nina; Horbańczuk, Jarosław Olav; Szumacher-Strabel, Małgorzata; Cieślak, Adam; Lipińska-Palka, Paulina; Józefiak, Damian; Kamińska, Agnieszka; Atanasov, Atanas G
2018-05-11
The aim of the study was to estimate the influence of the different levels of Cu, Zn, and Mn nanoparticles on the activity of aminopeptidases in turkey. An experiment was carried out on 144 turkey hen Hybrid Converter. The birds were divided into groups given standard- and nanoparticle-supplementation of different level of copper (Cu 20, 10, 2 mg/kg), zinc (Zn 100, 50, 10 ppm), and manganese (Mn 100, 50, 10 ppm), covering respectively 100%, 50%, and 10% of the physiological demands for those minerals in the diet. The activity of aminopeptidases (alanyl: AlaAP, leucyl: LeuAP and arginyl: ArgAP) after supplementation of minerals was determined in the breast and thigh turkey muscle. The strongest effect of interaction among minerals supplementation form and dose on the activity levels of the aminopeptidases in thigh muscle was observed for nano-Cu already at the lowest dose of 2 mg/kg. In this dose (covering 10% of the birds’ demand) nano form of supplementation significantly increased the activity of Ala-, Leu-, and ArgAP (877, 201, and 719, respectively), compared to standard form of supplementation (461, 90.5, and 576, respectively). In turn, in breast muscle, after supplementation covering 10% of the demand with the nano-Cu, nano-Zn, and nano-Mn compared to the standard form, we did not observe any significant difference in the activity levels of any of the investigated aminopeptidases, except for AlaAP under Zn supplementation. Supplementation with the 20 mg/kg of Nano-Cu (100% of demand) and with 10 mg/kg of Nano-Cu (50% of demand) inhibited the activity of all of the three aminopeptidases in thigh muscle. Supplementation of the minerals in nano form into the diet, especially of Cu and Zn in the dose covering 10% of the demand is relevant to maintain homeostasis in turkey muscles, as indicated by the activity of the aminopeptidases.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Gill, D.M.; Coburn, J.
1987-10-06
The authors have clarified relationships between cholera toxin, cholera toxin substrates, a membrane protein S that is required for toxin activity, and a soluble protein CF that is needed for the function of S. The toxin has little intrinsic ability to catalyze ADP-ribosylations unless it encounters the active form of the S protein, which is S liganded to GTP or to a GTP analogue. In the presence of CF, S x GTP forms readily, though reversibly, but a more permanent active species, S-guanosine 5'-O-(3-thiotriphosphate) (S x GTP..gamma..S), forms over a period of 10-15 min at 37/sup 0/C. Both guanosine 5'-O-(2-thiodiphosphate)more » and GTP block this quasi-permanent activation. Some S x GTP..gamma..S forms in membranes that are exposed to CF alone and then to GTP..gamma..S, with a wash in between, and it is possible that CF facilitates a G nucleotide exchange. S x GTP..gamma..S dissolved by nonionic detergents persists in solution and can be used to support the ADP-ribosylation of nucleotide-free substrates. In this circumstance, added guanyl nucleotides have no further effect. This active form of S is unstable, especially when heated, but the thermal inactivation above 45/sup 0/C is decreased by GTP..gamma..S. Active S is required equally for the ADP-ribosylation of all of cholera toxin's protein substrates, regardless of whether they bind GTP or not. They suggest that active S interacts directly with the enzymic A/sub 1/ fragments of cholera toxin and not with any toxin substrate. The activation and activity of S are independent of the state, or even the presence, of adenylate cyclase and seem to be involved with the cyclase system only via cholera toxin. S is apparently not related by function to certain other GTP binding proteins, including p21/sup ras/, and appears to be a new GTP binding protein whose physiologic role remains to be identified.« less
Nishigaki, Kazuo; Thompson, Delores; Hanson, Charlotte; Yugawa, Takashi; Ruscetti, Sandra
2001-01-01
The Friend spleen focus-forming virus (SFFV) encodes a unique envelope glycoprotein, gp55, which allows erythroid cells to proliferate and differentiate in the absence of erythropoietin (Epo). SFFV gp55 has been shown to interact with the Epo receptor complex, causing constitutive activation of various signal-transducing molecules. When injected into adult mice, SFFV induces a rapid erythroleukemia, with susceptibility being determined by the host gene Fv-2, which was recently shown to be identical to the gene encoding the receptor tyrosine kinase Stk/Ron. Susceptible, but not resistant, mice encode not only full-length Stk but also a truncated form of the kinase, sf-Stk, which may mediate the biological effects of SFFV infection. To determine whether expression of SFFV gp55 leads to the activation of sf-Stk, we expressed sf-Stk, with or without SFFV gp55, in hematopoietic cells expressing the Epo receptor. Our data indicate that sf-Stk interacts with SFFV gp55 as well as gp55P, the biologically active form of the viral glycoprotein, forming disulfide-linked complexes. This covalent interaction, as well as noncovalent interactions with SFFV gp55, results in constitutive tyrosine phosphorylation of sf-Stk and its association with multiple tyrosine-phosphorylated signal-transducing molecules. In contrast, neither Epo stimulation in the absence of SFFV gp55 expression nor expression of a mutant of SFFV that cannot interact with sf-Stk was able to induce tyrosine phosphorylation of sf-Stk or its association with any signal-transducing molecules. Covalent interaction of sf-Stk with SFFV gp55 and constitutive tyrosine phosphorylation of sf-Stk can also be detected in an erythroleukemia cell line derived from an SFFV-infected mouse. Our results suggest that SFFV gp55 may mediate its biological effects in vivo by interacting with and activating a truncated form of the receptor tyrosine kinase Stk. PMID:11483734
Olsen, Rikke K J; Andresen, Brage S; Christensen, Ernst; Bross, Peter; Skovby, Flemming; Gregersen, Niels
2003-07-01
Mutations in electron transfer flavoprotein (ETF) and its dehydrogenase (ETFDH) are the molecular basis of multiple acyl-CoA dehydrogenation deficiency (MADD), an autosomal recessively inherited and clinically heterogeneous disease that has been divided into three clinical forms: a neonatal-onset form with congenital anomalies (type I), a neonatal-onset form without congenital anomalies (type II), and a late-onset form (type III). To examine whether these different clinical forms could be explained by different ETF/ETFDH mutations that result in different levels of residual ETF/ETFDH enzyme activity, we have investigated the molecular genetic basis for disease development in nine patients representing the phenotypic spectrum of MADD. We report the genomic structures of the ETFA, ETFB, and ETFDH genes and the identification and characterization of seven novel and three previously reported disease-causing mutations. Our molecular genetic investigations of these nine patients are consistent with three clinical forms of MADD showing a clear relationship between the nature of the mutations and the severity of disease. Interestingly, our data suggest that homozygosity for two null mutations causes fetal development of congenital anomalies resulting in a type I disease phenotype. Even minute amounts of residual ETF/ETFDH activity seem to be sufficient to prevent embryonic development of congenital anomalies giving rise to type II disease. Overexpression studies of an ETFB-D128N missense mutation identified in a patient with type III disease showed that the residual activity of the mutant enzyme could be rescued up to 59% of that of wild-type activity when ETFB-D128N-transformed E. coli cells were grown at low temperature. This indicates that the effect of the ETF/ETFDH genotype in patients with milder forms of MADD, in whom residual enzyme activity allows modulation of the enzymatic phenotype, may be influenced by environmental factors like cellular temperature. Copyright 2003 Wiley-Liss, Inc.
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...; 086-0-27C, Coastal Analysis Form; 086-0-27D, Coastal Structures Form; 086-0-27E, Alluvial Fan Flooding... Analysis Form; 086-0-27D, Coastal Structures Form; 086-0-27E, Alluvial Fan Flooding Form. Abstract: The...-0-27E, Alluvial Fan 150 1 150 1 150 46.65 6,998 Flooding Form. Total 1,500 4,950 17,700 835,839...
Titov, V N
2014-02-01
In liver statins inhibit synthesis of specific pool of cholesterol which is formed de novo by hepatocytes for monolayer of polar lipids at the surface of forming lipoproteins of very low density. The statins, decreasing content of non-esterified cholesterol in monolayer, activate hydrolysis of triglycerides in lipoproteins of very low density, formation of lipoproteins of low density and their absorption by cells through apoB-100 receptors. The statins, activating absorption of lipoproteins of low density, restore functional action of essential polyenoic fatty acids. The essential polyenoic fatty acids, fibrates and glitazones form in cells effective oleic version of metabolism when mitochondrions predominantly oxidize oleic fatty acid. The statins, non-activating oxidation in peroxisomes and inhibiting activity of stearil-KoA-desaturase, form in cells less effective palmitic variant of metabolism of fatty acids under oxidation of palmitic fatty acid in mitochondrions. The fatty acids are not enough under hydrolysis of exogenous triglycerides to synthesize optimal amount of ATP. The fatty acids accumulated in adipocytes are to be used. This is the cause of formation by statins the resistance to insulin. Functionally, lipoproteins of very low density and lipoproteins of low density are phylogenetically different. The former ones transfer fatty acids to cells in the form of triglycerides and the latter ones--in the form of ethers with alcohol cholesterol. The statins normalize absorption of essential polyenoic fatty acids by cells which manifest a physiological action named a pleotropic one.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Tully, D.B.; Cidlowski, J.A.
1989-03-07
Sucrose density gradient shift assays were used to study the interactions of human glucocorticoid receptors (GR) with small DNA fragments either containing or lacking glucocorticoid response element (GRE) DNA consensus sequences. When crude cytoplasmic extracts containing ({sup 3}H)triamcinolone acetonide (({sup 3}H)TA) labeled GR were incubated with unlabeled DNA under conditions of DNA excess, a GRE-containing DNA fragment obtained from the 5' long terminal repeat of mouse mammary tumor virus (MMTV LTR) formed a stable 12-16S complex with activated, but not nonactivated, ({sup 3}H)TA receptor. By contrast, if the cytosols were treated with calf thymus DNA-cellulose to deplete non-GR-DNA-binding proteins priormore » to heat activation, a smaller 7-10S complex was formed with the MMTV LTR DNA fragment. Activated ({sup 3}H)TA receptor from DNA-cellulose pretreated cytosols also interacted with two similarly sized fragments from pBR322 DNA. Stability of the complexes formed between GR and these three DNA fragments was strongly affected by even moderate alterations in either the salt concentration or the pH of the gradient buffer. Under all conditions tested, the complex formed with the MMTV LTR DNA fragment was more stable than the complexes formed with either of the pBR322 DNA fragments. Together these observations indicate that the formation of stable complexes between activated GR and isolated DNA fragments requires the presence of GRE consensus sequences in the DNA.« less
75 FR 8407 - Agency Information Collection Activities: Proposed Collection; Comment Request
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... Request, NA Form 6006, which will be used by NARA employees, on-site contractors, volunteers, Foundation...: Identification Card Request. OMB number: 3095-0057. Agency form number: NA Form 6006. Type of review: Regular...
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A pore-forming protein implements VLR-activated complement cytotoxicity in lamprey.
Wu, Fenfang; Feng, Bo; Ren, Yong; Wu, Di; Chen, Yue; Huang, Shengfeng; Chen, Shangwu; Xu, Anlong
2017-01-01
Lamprey is a basal vertebrate with a unique adaptive immune system, which uses variable lymphocyte receptors (VLRs) for antigen recognition. Our previous study has shown that lamprey possessed a distinctive complement pathway activated by VLR. In this study, we identified a natterin family member-lamprey pore-forming protein (LPFP) with a jacalin-like lectin domain and an aerolysin-like pore-forming domain. LPFP had a high affinity with mannan and could form oligomer in the presence of mannan. LPFP could deposit on the surface of target cells, form pore-like complex resembling a wheel with hub and spokes, and mediate powerful cytotoxicity on target cells. These pore-forming proteins along with VLRs and complement molecules were essential for the specific cytotoxicity against exogenous pathogens and tumor cells. This unique cytotoxicity implemented by LPFP might emerge before or in parallel with the IgG-based classical complement lytic pathway completed by polyC9.
Paraoxonase activity and dementia.
Wehr, Hanna; Bednarska-Makaruk, Małgorzata; Graban, Ałła; Lipczyńska-Łojkowska, Wanda; Rodo, Maria; Bochyńska, Anna; Ryglewicz, Danuta
2009-08-15
Paraoxonase activity, homocysteine level and lipids were determined in 120 patients with dementia (51 with Alzheimer disease, 28 with dementia of vascular origin, 41 with mixed dementia), 45 with mild cognitive impairment and in 61 age and sex matched controls without dementia. Paraoxonase activity was decreased in Alzheimer disease and in mixed dementia as compared with control group. In the same forms of dementia homocysteine levels were increased. In Alzheimer disease paraoxonase activity was negatively correlated with homocysteine levels. Minimental State Examination results showed positive correlation with paraoxonase activity. The results suggest an important role of oxidative stress in the development of the forms of dementia with prevailing neurodegeneration.
Cost analysis in a CMHC: determining the cost of staff time.
Haring, A; Eckert, C
1979-06-01
The program evaluation and research unit of a community mental health center developed and field-tested a survey form to measure how employees spend their time. The form is divided into direct patient care activities, which include interviewing and testing, conducting therapy, and prescribing medications, and administrative or support activities, which include filling out charts, attending meetings, and training staff. All staff record daily, for one week, the hours and minutes they spend in each activity. Using that data as a base, the evaluation unit can determine the percentage of time staff spend in each activity and the cost of each activity based on staff members' paychecks.
Modeling Large-Scale Networks Using Virtual Machines and Physical Appliances
2014-01-27
downloaded and run locally. The lab solution couldn’t be based on ActiveX because the military Report Documentation Page Form ApprovedOMB No. 0704-0188...unclassified b. ABSTRACT unclassified c. THIS PAGE unclassified Standard Form 298 (Rev. 8-98) Prescribed by ANSI Std Z39-18 disallowed ActiveX support on...its systems, which made running an RDP client over ActiveX not possible. The challenges the SEI encountered in delivering the instruction were
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...-0-2A, National Fire Academy (NFA) Long-Term Evaluation Student/Trainee; FEMA Form 078-0-2, NFA Long... collection; OMB No. 1660-0039; FEMA Form 078-0-2A (Presently FEMA Form 95-59), NFA Long- Term Evaluation Student/Trainee; FEMA Form 078-0-2 (Presently FEMA Form 95-58), NFA Long-Term Evaluation Supervisors...
The purpose of this SOP is to define the steps for creating a scannable form and defining the database associated with the activated form. It applies to forms developed for the Arizona NHEXAS project and the Border study. Keywords: data; scannable forms.
The U.S.-Mexico Border...
21 CFR 1305.11 - Procedure for obtaining DEA Forms 222.
Code of Federal Regulations, 2011 CFR
2011-04-01
... 21 Food and Drugs 9 2011-04-01 2011-04-01 false Procedure for obtaining DEA Forms 222. 1305.11... I AND II CONTROLLED SUBSTANCES DEA Form 222 § 1305.11 Procedure for obtaining DEA Forms 222. (a) DEA..., which is based on the business activity of the registrant, will be imposed on the number of DEA Forms...
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... information collection; OMB No. 1660-0100; FEMA Forms 119-25-1 (replaces FEMA Form 75-5), General Admissions Application; 119-25-2 (replaces FEMA Form 75-5a), General Admissions Application Short-Form; 119-25-3 (replaces FEMA Form 75-3), Student Stipend Agreement; and 119-25-4 (replaces FEMA For 75-3a), Student...
Swearing, Euphemisms, and Linguistic Relativity
Bowers, Jeffrey S.; Pleydell-Pearce, Christopher W.
2011-01-01
Participants read aloud swear words, euphemisms of the swear words, and neutral stimuli while their autonomic activity was measured by electrodermal activity. The key finding was that autonomic responses to swear words were larger than to euphemisms and neutral stimuli. It is argued that the heightened response to swear words reflects a form of verbal conditioning in which the phonological form of the word is directly associated with an affective response. Euphemisms are effective because they replace the trigger (the offending word form) by another word form that expresses a similar idea. That is, word forms exert some control on affect and cognition in turn. We relate these findings to the linguistic relativity hypothesis, and suggest a simple mechanistic account of how language may influence thinking in this context. PMID:21799832
Sellés-Marchart, Susana; Casado-Vela, Juan; Bru-Martínez, Roque
2006-02-15
Polyphenol oxidase (PPO) has been extracted from both soluble and particulate fractions of loquat fruit (Eriobotrya japonica Lindl. cv. Algerie). The soluble PPO (20% of total activity) was partially purified 3.3-fold after ammonium sulfate fractionation being in its active state. The particulate PPO fraction (80% of total activity) was purified to homogeneity in a latent form being activable by sodium dodecyl sulfate (SDS). The enzyme was purified 40.0-fold with a total yield of 15.3% after extraction by phase partitioning in Triton X-114 followed by three chromatographic steps. The molecular weight was estimated to be about 59.2 and 61.2 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and gel filtration chromatography, respectively, indicating that latent PPO is a monomer. Latent PPO catalyzed the oxidation of chlorogenic acid (CA) at a rate 50-fold faster than that of 4-tert-butylcatechol (TBC) but the soluble active counterpart only twice. Both PPOs exhibited similar Km values for TBC but Km for CA was 5-fold higher for the latent than for the active soluble PPO. Other kinetic characteristics, including sensitivity to inhibitors, substrate specificity, thermal stability, temperature, and pH profiles, were quite different between both PPOs. These results provide strong evidences that the soluble active and the particulate latent are different forms of PPO in loquat fruit flesh. The results suggest that the major PPO form for the oxidation of CA, leading to enzymatic browning under physiological conditions, is the latent one.
1992-01-01
We analyzed the production of Torpedo marmorata acetylcholinesterase (AChE) in transfected COS cells. We report that the presence of an aspartic acid at position 397, homologous to that observed in other cholinesterases and related enzymes (Krejci, E., N. Duval, A. Chatonnet, P. Vincens, and J. Massoulie. 1991. Proc. Natl. Acad. Sci. USA. 88:6647-6651), is necessary for catalytic activity. The presence of an asparagine in the previously reported cDNA sequence (Sikorav, J.L., E. Krejci, and J. Massoulie. 1987. EMBO (Eur. Mol. Biol. Organ.) J. 6:1865-1873) was most likely due to a cloning error (codon AAC instead of GAC). We expressed the T and H subunits of Torpedo AChE, which differ in their COOH-terminal region and correspond respectively to the collagen-tailed asymmetric forms and to glycophosphatidylinositol-anchored dimers of Torpedo electric organs, as well as a truncated T subunit (T delta), lacking most of the COOH- terminal peptide. The transfected cells synthesized similar amounts of AChE immunoreactive protein at 37 degrees and 27 degrees C. However AChE activity was only produced at 27 degrees C and, even at this temperature, only a small proportion of the protein was active. We analyzed the molecular forms of active AChE produced at 27 degrees C. The H polypeptides generated glycophosphatidylinositol-anchored dimers, resembling the corresponding natural AChE form. The cells also released non-amphiphilic dimers G2na. The T polypeptides generated a series of active forms which are not produced in Torpedo electric organs: G1a, G2a, G4a, and G4na cellular forms and G2a and G4na secreted forms. The amphiphilic forms appeared to correspond to type II forms (Bon, S., J. P. Toutant, K. Meflah, and J. Massoulie. 1988. J. Neurochem. 51:776- 785; Bon, S., J. P. Toutant, K. Meflah, and J. Massoulie. 1988. J. Neurochem. 51:786-794), which are abundant in the nervous tissue and muscles of higher vertebrates (Bon, S., T. L. Rosenberry, and J. Massoulie. 1991. Cell. Mol. Neurobiol. 11:157-172). The H and T catalytic subunits are thus sufficient to account for all types of known AChE forms. The truncated T delta subunit yielded only non- amphiphilic monomers, demonstrating the importance of the T COOH- terminal peptide in the formation of oligomers, and in the hydrophobic character of type II forms. PMID:1639848
Active superconducting devices formed of thin films
Martens, Jon S.; Beyer, James B.; Nordman, James E.; Hohenwarter, Gert K. G.
1991-05-28
Active superconducting devices are formed of thin films of superconductor which include a main conduction channel which has an active weak link region. The weak link region is composed of an array of links of thin film superconductor spaced from one another by voids and selected in size and thickness such that magnetic flux can propagate across the weak link region when it is superconducting. Magnetic flux applied to the weak link region will propagate across the array of links causing localized loss of superconductivity in the links and changing the effective resistance across the links. The magnetic flux can be applied from a control line formed of a superconducting film deposited coplanar with the main conduction channel and weak link region on a substrate. The devices can be formed of any type to superconductor but are particularly well suited to the high temperature superconductors since the devices can be entirely formed from coplanar films with no overlying regions. The devices can be utilized for a variety of electrical components, including switching circuits, amplifiers, oscillators and modulators, and are well suited to microwave frequency applications.
Radjainia, Mazdak; Venugopal, Hariprasad; Desfosses, Ambroise; Phillips, Amy J; Yewdall, N Amy; Hampton, Mark B; Gerrard, Juliet A; Mitra, Alok K
2015-05-05
Peroxiredoxins (Prxs) are a ubiquitous class of thiol-dependent peroxidases that play an important role in the protection and response of cells to oxidative stress. The catalytic unit of typical 2-Cys Prxs are homodimers, which can self-associate to form complex assemblies that are hypothesized to have signaling and chaperone activity. Mitochondrial Prx3 forms dodecameric toroids, which can further stack to form filaments, the so-called high-molecular-weight (HMW) form that has putative holdase activity. We used single-particle analysis and helical processing of electron cryomicroscopy images of human Prx3 filaments induced by low pH to generate a ∼7-Å resolution 3D structure of the HMW form, the first such structure for a 2-Cys Prx. The pseudo-atomic model reveals interactions that promote the stacking of the toroids and shows that unlike previously reported data, the structure can accommodate a partially folded C terminus. The HMW filament lumen displays hydrophobic patches, which we hypothesize bestow holdase activity. Copyright © 2015 Elsevier Ltd. All rights reserved.
Julesz, Máté
2014-08-10
The passive form of euthanasia is legalized almost in every civilized country. Its active form is not a generally accepted legal institution. In Europe, active euthanasia is legalized only in The Netherlands, Belgium, Luxembourg and Switzerland. In Australia, the Act on the Rights of the Terminally Ill of 1995 legalized the institution of assisted suicide, which is not identical to active euthanasia. The difference lies in the fact that legalized active euthanasia means that the author of a murder is not punishable (under certain circumstances), whilst assisted suicide is not about murder, rather about suicide. In the first case, the patient is killed on his or her request by someone else. In the second case, the patient himself or herself executes the act of self-killing (by the assistance of a healthcare worker). In Australia, the institution of assisted suicide was repealed in 1997. Assisted suicide is legal in four USA member states: in Vermont, Washington, Montana and Oregon. In Uruguay, the active form of euthanasia has been legal since 1932.
NASA Astrophysics Data System (ADS)
Huang, Haiping
2017-03-01
To understand the collective spiking activity in neuronal populations, it is essential to reveal basic circuit variables responsible for these emergent functional states. Here, I develop a mean field theory for the population coupling recently proposed in the studies of the visual cortex of mouse and monkey, relating the individual neuron activity to the population activity, and extend the original form to the second order, relating neuron-pair’s activity to the population activity, to explain the high order correlations observed in the neural data. I test the computational framework on the salamander retinal data and the cortical spiking data of behaving rats. For the retinal data, the original form of population coupling and its advanced form can explain a significant fraction of two-cell correlations and three-cell correlations, respectively. For the cortical data, the performance becomes much better, and the second order population coupling reveals non-local effects in local cortical circuits.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Wang Lei; Sasai, Ken; Akagi, Tsuyoshi
2008-08-29
The AKT pathway is frequently activated in glioblastoma, and as such, inhibitors of this pathway could prove very useful as anti-glioblastoma therapies. Here we established immortalized astrocytes expressing Renilla luciferase as well as those expressing both an active form of AKT and firefly luciferase. Since both luciferase activities represent the numbers of corresponding cell lines, novel inhibitors of the AKT pathway can be identified by treating co-cultures containing the two types of luciferase-expressing cells with individual compounds. Indeed, such a screening system succeeded in identifying fumitremorgin C as an efficient inhibitor of the AKT pathway, which was further confirmed bymore » the ability of fumitremorgin C to selectively inhibit the growth of immortalized astrocytes expressing an active form of AKT. The present study proposes a broadly applicable approach for identifying therapeutic agents that target the pathways and/or molecules responsible for cancer development.« less
DOE Office of Scientific and Technical Information (OSTI.GOV)
Hagvall, Lina; Baron, Jens Malte; Boerje, Anna
2008-12-01
Contact sensitization is caused by low molecular weight compounds which penetrate the skin and bind to protein. In many cases, these compounds are activated to reactive species, either by autoxidation on exposure to air or by metabolic activation in the skin. Geraniol, a widely used fragrance chemical, is considered to be a weak allergen, although its chemical structure does not indicate it to be a contact sensitizer. We have shown that geraniol autoxidizes and forms allergenic oxidation products. In the literature, it is suggested but not shown that geraniol could be metabolically activated to geranial. Previously, a skin-like CYP cocktailmore » consisting of cutaneous CYP isoenzymes, was developed as a model system to study cutaneous metabolism. In the present study, we used this system to investigate CYP-mediated activation of geraniol. In incubations with the skin-like CYP cocktail, geranial, neral, 2,3-epoxygeraniol, 6,7-epoxygeraniol and 6,7-epoxygeranial were identified. Geranial was the main metabolite formed followed by 6,7-epoxygeraniol. The allergenic activities of the identified metabolites were determined in the murine local lymph node assay (LLNA). Geranial, neral and 6,7-epoxygeraniol were shown to be moderate sensitizers, and 6,7-epoxygeranial a strong sensitizer. Of the isoenzymes studied, CYP2B6, CYP1A1 and CYP3A5 showed high activities. It is likely that CYP1A1 and CYP3A5 are mainly responsible for the metabolic activation of geraniol in the skin, as they are expressed constitutively at significantly higher levels than CYP2B6. Thus, geraniol is activated through both autoxidation and metabolism. The allergens geranial and neral are formed via both oxidation mechanisms, thereby playing a large role in the sensitization to geraniol.« less
Hagvall, Lina; Baron, Jens Malte; Börje, Anna; Weidolf, Lars; Merk, Hans; Karlberg, Ann-Therese
2008-12-01
Contact sensitization is caused by low molecular weight compounds which penetrate the skin and bind to protein. In many cases, these compounds are activated to reactive species, either by autoxidation on exposure to air or by metabolic activation in the skin. Geraniol, a widely used fragrance chemical, is considered to be a weak allergen, although its chemical structure does not indicate it to be a contact sensitizer. We have shown that geraniol autoxidizes and forms allergenic oxidation products. In the literature, it is suggested but not shown that geraniol could be metabolically activated to geranial. Previously, a skin-like CYP cocktail consisting of cutaneous CYP isoenzymes, was developed as a model system to study cutaneous metabolism. In the present study, we used this system to investigate CYP-mediated activation of geraniol. In incubations with the skin-like CYP cocktail, geranial, neral, 2,3-epoxygeraniol, 6,7-epoxygeraniol and 6,7-epoxygeranial were identified. Geranial was the main metabolite formed followed by 6,7-epoxygeraniol. The allergenic activities of the identified metabolites were determined in the murine local lymph node assay (LLNA). Geranial, neral and 6,7-epoxygeraniol were shown to be moderate sensitizers, and 6,7-epoxygeranial a strong sensitizer. Of the isoenzymes studied, CYP2B6, CYP1A1 and CYP3A5 showed high activities. It is likely that CYP1A1 and CYP3A5 are mainly responsible for the metabolic activation of geraniol in the skin, as they are expressed constitutively at significantly higher levels than CYP2B6. Thus, geraniol is activated through both autoxidation and metabolism. The allergens geranial and neral are formed via both oxidation mechanisms, thereby playing a large role in the sensitization to geraniol.
Updated Liquid Secondary Waste Grout Formulation and Preliminary Waste Form Qualification
DOE Office of Scientific and Technical Information (OSTI.GOV)
Saslow, Sarah A.; Um, Wooyong; Russell, Renee L.
This report describes the results from liquid secondary waste grout (LSWG) formulation and cementitious waste form qualification tests performed by Pacific Northwest National Laboratory (PNNL) for Washington River Protection Solutions, LLC (WRPS). New formulations for preparing a cementitious waste form from a high-sulfate liquid secondary waste stream simulant, developed for Effluent Management Facility (EMF) process condensates merged with low activity waste (LAW) caustic scrubber, and the release of key constituents (e.g. 99Tc and 129I) from these monoliths were evaluated. This work supports a technology development program to address the technology needs for Hanford Site Effluent Treatment Facility (ETF) liquid secondarymore » waste (LSW) solidification and supports future Direct Feed Low-Activity Waste (DFLAW) operations. High-priority activities included simulant development, LSWG formulation, and waste form qualification. The work contained within this report relates to waste form development and testing and does not directly support the 2017 integrated disposal facility (IDF) performance assessment (PA). However, this work contains valuable information for use in PA maintenance past FY17, and for future waste form development efforts. The provided data should be used by (i) cementitious waste form scientists to further understanding of cementitious dissolution behavior, (ii) IDF PA modelers who use quantified constituent leachability, effective diffusivity, and partitioning coefficients to advance PA modeling efforts, and (iii) the U.S. Department of Energy (DOE) contractors and decision makers as they assess the IDF PA program. The results obtained help fill existing data gaps, support final selection of a LSWG waste form, and improve the technical defensibility of long-term waste form performance estimates.« less
[Consideration of drug absorption in customizing drug therapy].
Walter-Sack, I; Haefeli, W E
2000-09-01
The rate and extent of drug absorption from the small intestine are related to the release of the active ingredient from a dosage form, its solubility in the liquid phase of gastrointestinal contents, and the transport of the dissolved compound or the intact dosage form from the stomach into the duodenum. With pharmaceutical preparations releasing the active compound within the stomach, and enteric-coated "micro"-formulations (micropellets), gastric emptying is possible during the interdigestive and the digestive period. Potential differences of drug absorption between fasting administration and intake during the digestive period are unpredictable, because they are related to the release characteristics of the dosage form. However, larger enteric-coated preparations like tablets can leave the stomach only with a phase 3 contraction of fasting motility; intake during the digestive period will result in gastric retention of this type of dosage form until all food has left the stomach and fasting motility is restored. Consequently the onset of drug absorption is delayed. This interaction between food and large enteric-coated dosage forms is predictable from pyloric function in relation to the gastric motility. As it occurs regularly, it can be taken into account when prescribing enteric-coated dosage forms. If concomitant intake of food and enteric-coated drugs is unavoidable, but a rapid onset of drug absorption is necessary, micropellets are the dosage form of choice. When the therapeutic effect is insufficient, drug dosage form and timing of drug administration should be checked before prescribing a different active compound.
Wu, Xiaoyang; Katz, Evan; Valle, Maria Cecilia Della; Mascioli, Kirsten; Flanagan, John J; Castelli, Jeffrey P; Schiffmann, Raphael; Boudes, Pol; Lockhart, David J; Valenzano, Kenneth J; Benjamin, Elfrida R
2011-01-01
Fabry disease is caused by mutations in the gene (GLA) that encodes α-galactosidase A (α-Gal A). The iminosugar AT1001 (GR181413A, migalastat hydrochloride, 1-deoxygalactonojirimycin) is a pharmacological chaperone that selectively binds and stabilizes α-Gal A, increasing total cellular levels and activity for some mutant forms (defined as “responsive”). In this study, we developed a cell-based assay in cultured HEK-293 cells to identify mutant forms of α-Gal A that are responsive to AT1001. Concentration-dependent increases in α-Gal A activity in response to AT1001 were shown for 49 (60%) of 81 mutant forms. The responses of α-Gal A mutant forms were generally consistent with the responses observed in male Fabry patient-derived lymphoblasts. Importantly, the HEK-293 cell responses of 19 α-Gal A mutant forms to a clinically achievable concentration of AT1001 (10 µM) were generally consistent with observed increases in α-Gal A activity in peripheral blood mononuclear cells from male Fabry patients orally administered AT1001 during Phase 2 clinical studies. This indicates that the cell-based responses can identify mutant forms of α-Gal A that are likely to respond to AT1001 in vivo. Thus, the HEK-293 cell-based assay may be a useful aid in the identification of Fabry patients with AT1001-responsive mutant forms. Hum Mutat 32:1–13, 2011. © 2011 Wiley-Liss, Inc. PMID:21598360
Perlstein, Deborah L.; Ge, Jie; Ortigosa, Allison D.; Robblee, John H.; Zhang, Zhen; Huang, Mingxia; Stubbe, JoAnne
2015-01-01
The class I ribonucleotide reductases (RNRs) are composed of two homodimeric subunits: R1 and R2. R2 houses a diferric-tyrosyl radical (Y•) cofactor. Saccharomyces cerevisiae has two R2s: Y2 (β2) and Y4 (β′2). Y4 is an unusual R2 because three residues required for iron binding have been mutated. While the heterodimer (ββ′) is thought to be the active form, several rnr4Δ strains are viable. To resolve this paradox, N-terminally epitope-tagged β and β′ were expressed in E. coli or integrated into the yeast genome. In vitro exchange studies reveal that when apo-(His6)-β2 (Hisβ2) is mixed with β′2, apo-Hisββ′ forms quantitatively within 2 min. In contrast, holo-ββ′ fails to exchange with apo-Hisβ2 to form holo-Hisββ and β′2. Isolation of genomically encoded tagged β or β′ from yeast extracts gave a 1:1 complex of β and β′, suggesting that ββ′ is the active form. The catalytic activity, protein concentrations, and Y• content of the rnr4Δ and wild type (wt) strains were compared to clarify the role of β′ in vivo. The Y• content of rnr4Δ is 15-fold less than that of wt, consistent with the observed low activity of rnr4Δ extracts (<0.01 nmol min−1 mg−1) versus wt (0.06 ± 0.01 nmol min−1 mg−1). FLAGβ2 isolated from the rnr4Δ strain has a specific activity of 2 nmol min−1 mg−1, similar to that of reconstituted apo-Hisβ2 (10 nmol min−1 mg−1), but significantly less than holo-Hisββ′ (~2000 nmol min−1 mg−1). These studies together demonstrate that β′ plays a crucial role in cluster assembly in vitro and in vivo and that the active form of the yeast R2 is ββ′. PMID:16285741
75 FR 57480 - Agency Information Collection Activities: Customs Declaration (Form 6059B)
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Molitor, Christian; Mauracher, Stephan Gerhard; Rompel, Annette, E-mail: annette.rompel@univie.ac.at
2015-05-22
Latent and active aurone synthase purified from petals of C. grandiflora (cgAUS1) were crystallized. The crystal quality of recombinantly expressed latent cgAUS1 was significantly improved by co-crystallization with the polyoxotungstate Na{sub 6}[TeW{sub 6}O{sub 24}] within the liquid–liquid phase-separation zone. Aurone synthase (AUS), a member of a novel group of plant polyphenol oxidases (PPOs), catalyzes the oxidative conversion of chalcones to aurones. Two active cgAUS1 (41.6 kDa) forms that differed in the level of phosphorylation or sulfation as well as the latent precursor form (58.9 kDa) were purified from the petals of Coreopsis grandiflora. The differing active cgAUS1 forms and themore » latent cgAUS1 as well as recombinantly expressed latent cgAUS1 were crystallized, resulting in six different crystal forms. The active forms crystallized in space groups P2{sub 1}2{sub 1}2{sub 1} and P12{sub 1}1 and diffracted to ∼1.65 Å resolution. Co-crystallization of active cgAUS1 with 1,4-resorcinol led to crystals belonging to space group P3{sub 1}21. The crystals of latent cgAUS1 belonged to space group P12{sub 1}1 and diffracted to 2.50 Å resolution. Co-crystallization of recombinantly expressed pro-AUS with the hexatungstotellurate(VI) salt Na{sub 6}[TeW{sub 6}O{sub 24}] within the liquid–liquid phase separation zone significantly improved the quality of the crystals compared with crystals obtained without hexatungstotellurate(VI)« less
Celedón, Gloria; González, Gustavo; Gulppi, Felipe; Pazos, Fabiola; Lanio, María E; Alvarez, Carlos; Calderón, Cristian; Montecinos, Rodrigo; Lissi, Eduardo
2013-12-01
Sticholysin II (St II) is a haemolytic toxin isolated from the sea anemone Stichodactyla helianthus. The high haemolytic activity of this toxin is strongly dependent on the red cell status and the macromolecule conformation. In the present communication we evaluate the effect of human serum albumin on St II haemolytic activity and its capacity to form pores in the bilayer of synthetic liposomes. St II retains its pore forming capacity in the presence of large concentrations (up to 500 μM) of human serum albumin. This effect is observed both in its capacity to produce red blood cells haemolysis and to generate functional pores in liposomes. In particular, the capacity of the toxin to lyse red blood cells increases in the presence of human serum albumin (HSA). Regarding the rate of the pore forming process, it is moderately decreased in liposomes and in red blood cells, in spite of an almost total coverage of the interface by albumin. All the data obtained in red cells and model membranes show that St II remains lytically active even in the presence of high HSA concentrations. This stubbornness can explain why the toxin is able to exert its haemolytic activity on membranes immersed in complex plasma matrixes such as those present in living organisms.
Janciauskiene, S
2001-03-26
Serine proteinase inhibitors (Serpins) are irreversible suicide inhibitors of proteases that regulate diverse physiological processes such as coagulation, fibrinolysis, complement activation, angiogenesis, apoptosis, inflammation, neoplasia and viral pathogenesis. The molecular structure and physical properties of serpins permit these proteins to adopt a number of variant conformations under physiological conditions including the native inhibitory form and several inactive, non-inhibitory forms, such as complexes with protease or other ligands, cleaved, polymerised and oxidised. Alterations of a serpin which affect its structure and/or secretion and thus reduce its functional levels may result in pathology. Serpin dysfunction has been implicated in thrombosis, emphysema, liver cirrhosis, immune hypersensitivity and mental disorders. The loss of inhibitory activity of serpins necessarily results in an imbalance between proteases and their inhibitors, but it may also have other physiological effects through the generation of abnormal concentrations of modified, non-inhibitory forms of serpins. Although these forms of inhibitory serpins are detected in tissues and fluids recovered from inflammatory sites, the important questions of which conditions result in generation of different molecular forms of serpins, what biological function these forms have, and which of them are directly linked to pathologies and/or may be useful markers for characterisation of disease states, remain to be answered. Elucidation of the biological activities of non-inhibitory forms of serpins may provide useful insights into the pathogenesis of diseases and suggest new therapeutic strategies.
Spezzati, Giulia; Su, Yaqiong; Hofmann, Jan P.; ...
2017-09-07
Ceria-supported Pd is a promising heterogeneous catalyst for CO oxidation relevant to environmental cleanup reactions. Pd loaded onto a nanorod form of ceria exposing predominantly (111) facets is already active at 50 °C. Here we report a combination of CO-FTIR spectroscopy and theoretical calculations that allows assigning different forms of Pd on the CeO 2(111) surface during reaction conditions. Single Pd atoms stabilized in the form of PdO and PdO 2 in a CO/O 2 atmosphere participate in a catalytic cycle involving very low activation barriers for CO oxidation. In conclusion, the presence of single Pd atoms on the Pd/CeOmore » 2-nanorod, corroborated by aberration-corrected TEM and CO-FTIR spectroscopy, is considered pivotal to its high CO oxidation activity.« less
Tocopherols in cancer: an update
Gupta, Soumyasri Das; Suh, Nanjoo
2016-01-01
Tocopherols exist in four forms designated as α, β, δ and γ. Due to their strong antioxidant properties, tocopherols have been suggested to reduce the risk of cancer. Cancer prevention studies with tocopherols have mostly utilized α-tocopherol. Large scale clinical trials with α-tocopherol provided inconsistent results regarding the cancer preventive activities of tocopherols. This review summarizes our current understanding of the anti-cancer activities of different forms of tocopherols based on follow up of the clinical trials, recent epidemiological evidences and experimental studies using in vitro and in vivo models. The experimental data provide strong evidence in support of the anti-cancer activities of δ-tocopherol,γ-tocopherol and the natural tocopherol mixture rich in γ-tocopherol, γ-TmT, over α-tocopherol. Such outcomes emphasize the need for detailed investigation into the cancer preventive activities of different forms of tocopherols to provide a strong rationale for intervention studies in the future. PMID:26751721
DOE Office of Scientific and Technical Information (OSTI.GOV)
Spezzati, Giulia; Su, Yaqiong; Hofmann, Jan P.
Ceria-supported Pd is a promising heterogeneous catalyst for CO oxidation relevant to environmental cleanup reactions. Pd loaded onto a nanorod form of ceria exposing predominantly (111) facets is already active at 50 °C. Here we report a combination of CO-FTIR spectroscopy and theoretical calculations that allows assigning different forms of Pd on the CeO 2(111) surface during reaction conditions. Single Pd atoms stabilized in the form of PdO and PdO 2 in a CO/O 2 atmosphere participate in a catalytic cycle involving very low activation barriers for CO oxidation. In conclusion, the presence of single Pd atoms on the Pd/CeOmore » 2-nanorod, corroborated by aberration-corrected TEM and CO-FTIR spectroscopy, is considered pivotal to its high CO oxidation activity.« less
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2012-03-13
...-Day Notice of Information Collection Under Review: Form I- 829, Petition by Entrepreneur to Remove... existing information collection. (2) Title of the Form/Collection: Petition by Entrepreneur to Remove... households. This form is used by a conditional resident alien entrepreneur who obtained such status through a...
Federal Register 2010, 2011, 2012, 2013, 2014
2011-03-08
... ACTION: 30-Day Notice of Information Collection Under Review: Form I- 829, Petition by Entrepreneur to... the Form/Collection: Petition by Entrepreneur to Remove Conditions. (3) Agency form number, if any... conditional resident alien entrepreneur who obtained such status through a qualifying investment, to apply to...
Federal Register 2010, 2011, 2012, 2013, 2014
2010-11-30
... ACTION: 60-Day notice of information collection under review: Form I- 829, Petition by Entrepreneur to... existing information collection. (2) Title of the Form/Collection: Petition by Entrepreneur to Remove... households. This form is used by a conditional resident alien entrepreneur who obtained such status through a...
Federal Register 2010, 2011, 2012, 2013, 2014
2012-06-07
... ACTION: 60-Day Notice of Information Collection Under Review: Form I- 829, Petition by Entrepreneur to...: Extension of an existing information collection. (2) Title of the Form/Collection: Petition by Entrepreneur...: Individuals and households. This form is used by a conditional resident alien entrepreneur who obtained such...
Federal Register 2010, 2011, 2012, 2013, 2014
2011-06-08
... Management Sub-grant; FEMA Form 078-0-2 (previously FEMA Form 90-32), Principal Advisor's Report. SUMMARY...), Request for Fire Management Sub-grant; FEMA Form 078-0-2 (previously FEMA Form 90-32), Principal Advisor's... DEPARTMENT OF HOMELAND SECURITY Federal Emergency Management Agency [Docket ID: FEMA-2011-0003...
Federal Register 2010, 2011, 2012, 2013, 2014
2011-03-31
... DEPARTMENT OF HOMELAND SECURITY United States Immigration and Customs Enforcement Agency... and Local Agency Assessment; OMB Control No. 1653-0040. The Department of Homeland Security, U.S... Security sponsoring the collection: Form 70-003, Form 70-004, Form 75-001 and Form 75-002; U.S. Immigration...
La forme en -l en macedonien (The Form Suffixed -1 in Macedonian).
ERIC Educational Resources Information Center
Hristova, Doreana
The Macedonian verb form corresponding to the form ending in "-l" in French is examined, focusing on the active past participle, which represents the past indeterminate or non-testimonial tense. Special attention is paid to aspectual, modal, temporal, and prosodic values, and all examples are drawn from the two languages. (MSE)
Federal Register 2010, 2011, 2012, 2013, 2014
2013-10-02
... (Statement of Dependency of Parent(s)) Activity Under OMB Review AGENCY: Veterans Benefits Administration... INFORMATION: Title: Statement of Dependency of Parent(s), VA Form 21-509. OMB Control Number: 2900-0089. Type...) for support complete VA Form 21-509 to report income and dependency information. Surviving parents of...
Federal Register 2010, 2011, 2012, 2013, 2014
2013-12-18
... Collection Activities: Proposed Collection; Comments Requested: Reports of Suspicious Orders or Theft/Loss of... Suspicious Orders or Theft/Loss of Listed Chemicals/Machines. (3) Agency form number, if any, and the... suspicious orders and thefts is provided in writing on an as needed basis and does not occur using a form...
Federal Register 2010, 2011, 2012, 2013, 2014
2012-06-27
... of Lapse--Government Life Insurance) Activities Under OMB Review AGENCY: Veterans Benefits...--Government Life Insurance, VA Form 29-389. b. Application for Reinstatement, VA Form 29-389-1. OMB Control...-389 and 29-389-1 are used to inform claimants that their government life insurance has lapsed or will...
Federal Register 2010, 2011, 2012, 2013, 2014
2011-07-25
... Activities: Arrival and Departure Record (Forms I-94 and I-94W) and Electronic System for Travel... I-94W (Nonimmigrant Visa Waiver Arrival/Departure), and the Electronic System for Travel... points: (1) Evaluate whether the proposed collection of information is necessary for the proper...
ERIC Educational Resources Information Center
Safadi, Rafi; Safadi, Ekhlass; Meidav, Meir
2017-01-01
This study compared students' learning in troubleshooting and problem solving activities. The troubleshooting activities provided students with solutions to conceptual problems in the form of refutation texts; namely, solutions that portray common misconceptions, refute them, and then present the accepted scientific ideas. They required students…
ERIC Educational Resources Information Center
Çekmez, Erdem
2016-01-01
This study investigates the effectiveness of a teaching activity that aimed to convey the meaning of indeterminate forms to a group of undergraduate students who were enrolled in an elementary mathematics education programme. The study reports the implementation sequence of the activity and students' experiences in the classroom. To assess the…
Development of immunoassays for human urokinase
NASA Technical Reports Server (NTRS)
Atassi, M. Zouhair
1988-01-01
Radioimmune assays (RIA) and enzyme linked immune assays for measurement of pro-urokinase and the two active forms of the enzyme were developed. Polyclonal and monoclonal antibodies, with desired specificities against preselected synthetic regions of urokinase (UK), were obtained by immunization with the respective synthetic peptides and used to develop RIA for zymogen and the two activated forms of UK.
30 CFR 580.12 - What must I include in my application or notification?
Code of Federal Regulations, 2012 CFR
2012-07-01
... permit application form (Form BOEM-0134) at least 30 days before the startup date for activities in the... type, location, purpose, and dates of activity; and environmental and other information. A... type of research and a brief description of how you will conduct it; (4) A map, plat, or chart, that...
30 CFR 580.12 - What must I include in my application or notification?
Code of Federal Regulations, 2014 CFR
2014-07-01
... permit application form (Form BOEM-0134) at least 30 days before the startup date for activities in the... type, location, purpose, and dates of activity; and environmental and other information. A... type of research and a brief description of how you will conduct it; (4) A map, plat, or chart, that...
30 CFR 580.12 - What must I include in my application or notification?
Code of Federal Regulations, 2013 CFR
2013-07-01
... permit application form (Form BOEM-0134) at least 30 days before the startup date for activities in the... type, location, purpose, and dates of activity; and environmental and other information. A... type of research and a brief description of how you will conduct it; (4) A map, plat, or chart, that...
Code of Federal Regulations, 2010 CFR
2010-01-01
... PERSONNEL MANAGEMENT (CONTINUED) CIVIL SERVICE REGULATIONS (CONTINUED) POLITICAL ACTIVITIES OF FEDERAL... the payroll employees who would process or administer such forms. However, the employee may mail his or her direct deposit form to his or her agency payroll office. (b) [Reserved] [59 FR 48769, Sept. 23...
Federal Register 2010, 2011, 2012, 2013, 2014
2010-12-16
... (Application for Reimbursement of National Test Fee) Activity Under OMB Review AGENCY: Veterans Benefits... Reimbursement of National Test Fee, VA Form 22-0810. OMB Control Number: 2900-0706. Type of Review: Extension of... complete VA Form 22-0810 to request reimbursement of national test fees. VA will use the data collected to...
Federal Register 2010, 2011, 2012, 2013, 2014
2013-03-01
... DEPARTMENT OF THE INTERIOR U.S. Geological Survey [USGS-GX13LR000F60100] Agency Information Collection Activities: Comment Request for the Production Estimate (2 Forms) AGENCY: U.S. Geological Survey... Officer, U.S. Geological Survey, 12201 Sunrise Valley Drive, Mail Stop 807, Reston, VA 20192 (mail); 703...
Federal Register 2010, 2011, 2012, 2013, 2014
2011-07-22
... Purchase and Contract of Sale) Activity Under OMB Review AGENCY: Veterans Benefits Administration... Management and Budget (OMB) for review and comment. The PRA submission describes the nature of the... Purchase and Contract of Sale, VA Form 26-6705. b. Credit Statement of Prospective Purchaser, VA Form 26...
Treatability of Aqueous Film-Forming Foams Used for Fire Fighting.
BIODETERIORATION, *FIRE EXTINGUISHING AGENTS, SURFACE ACTIVE SUBSTANCES, FLUORINATED HYDROCARBONS, FOAM , ACTIVATED SLUDGE PROCESS, ACTIVATED CARBON, TOXICITY, WASTE DISPOSAL, TABLES(DATA), ADSORPTION.