Science.gov

Sample records for partially irreversible inactivation

  1. Peroxidatic degradation of azide by catalase and irreversible enzyme inactivation.

    PubMed

    Lardinois, O M; Rouxhet, P G

    1996-12-01

    A study of the azide reaction with bovine liver catalase in presence of hydrogen peroxide has been performed, using conventional UV-visible spectrometry and activity measurements. Compound III and NO-ferrocatalase were the predominant forms of the enzyme observed in air and under nitrogen, respectively. A reaction scheme for peroxidatic degradation of azide by catalase is proposed. Accordingly, accumulation of Compound III is the main factor responsible for the reversible inhibition of 'catalatic' activity by azide, while formation of a complex between native catalase and azide has a negligible effect. Catalase is irreversibly inactivated by prolonged exposure to high levels of H2O2 and azide. The latter involves cleavage of the prosthetic group with liberation of the heme iron. Both in air and under nitrogen, generation of azidyl radicals seems to play a minor role in the irreversible inactivation process. PMID:8980644

  2. Partially-irreversible sorption of formaldehyde in five polymers

    NASA Astrophysics Data System (ADS)

    Ye, Wei; Cox, Steven S.; Zhao, Xiaomin; Frazier, Charles E.; Little, John C.

    2014-12-01

    Due to its environmental ubiquity and concern over its potential toxicity, the mass-transfer characteristics of formaldehyde are of critical importance to indoor air quality research. Previous studies have suggested that formaldehyde mass transfer in polymer is partially irreversible. In this study, mechanisms that could cause the observed irreversibility were investigated. Polycarbonate and four other polymeric matrices were selected and subjected to formaldehyde sorption/desorption cycles. Mass transfer of formaldehyde was partially irreversible in all cases, and three potential mechanisms were evaluated. First, attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) analysis was used to investigate possible formaldehyde polymerization on polymer surfaces. ATR-FTIR showed no detectable paraformaldehyde or formaldehyde on the film surfaces that had been exposed to formaldehyde and air. ATR-FTIR did detect aliphatic acids suggesting oxidation had occurred on film surfaces as a result of exposure to formaldehyde. However, additional study suggested that air is not the primary cause for irreversibility. Second, statistical physics theory was tested as a possible explanation. According to this theory, reversible and irreversible sorption could be taking place simultaneously. The irreversible fraction should be constant during sorption and the fraction could be determined by performing a complete sorption/desorption test. The sorption/desorption data was consistent with this theory. Third, chemisorption was considered as another possible cause for irreversibility. Extraction/fluorimetry testing of post-sorption and post-desorption polymer films showed measurable quantities of formaldehyde suggesting that some of the chemisorbed formaldehyde was reversible at the higher extraction temperature. Further quantitative study on chemical reaction products is needed.

  3. Partial inactivation of cytochrome c oxidase by nonpolar mercurial reagents

    SciTech Connect

    Mann, A.J.; Auer, H.E.

    1980-01-25

    Purified beef heart cytochrome c oxidase is inactivated to the extent of 35 to 50% by the nonpolar mercurial reagents mercuric chloride and ethylmercuric chloride. The inactivation is complete within 5 min. In titrations of activity, the plateau level of inactivation is attained at added ethylmercuric chloride:heme a ratios of about 1:1. Up to 3 mercury atoms/heme a are bound to the oxidase, although only the first of these affects its enzymatic activity. Incubation of the ethylmercury-modified oxidase with sulfhydryl compounds reverses the inactivation, with 2,3-dimercaptopropanol being most effective of the reagents tested. Spectrophotometric and polarographic assays of enzymatic activity show that K/sub m/ values for the native and the ethylmercury-modified enzymes are practically indistinguishable, and that the partial inactivation observed for the latter is reflected exclusively in a lower value of V/sub max/ compared to that of the native enzyme. Based on these results, we propose that ethylmercuric chloride reacts with a single crucial--SH group per heme a, and that electron transfer processes in the modified product are partially inhibited.

  4. Reversible, partial inactivation of plant betaine aldehyde dehydrogenase by betaine aldehyde: mechanism and possible physiological implications.

    PubMed

    Zárate-Romero, Andrés; Murillo-Melo, Darío S; Mújica-Jiménez, Carlos; Montiel, Carmina; Muñoz-Clares, Rosario A

    2016-04-01

    In plants, the last step in the biosynthesis of the osmoprotectant glycine betaine (GB) is the NAD(+)-dependent oxidation of betaine aldehyde (BAL) catalysed by some aldehyde dehydrogenase (ALDH) 10 enzymes that exhibit betaine aldehyde dehydrogenase (BADH) activity. Given the irreversibility of the reaction, the short-term regulation of these enzymes is of great physiological relevance to avoid adverse decreases in the NAD(+):NADH ratio. In the present study, we report that the Spinacia oleracea BADH (SoBADH) is reversibly and partially inactivated by BAL in the absence of NAD(+)in a time- and concentration-dependent mode. Crystallographic evidence indicates that the non-essential Cys(450)(SoBADH numbering) forms a thiohemiacetal with BAL, totally blocking the productive binding of the aldehyde. It is of interest that, in contrast to Cys(450), the catalytic cysteine (Cys(291)) did not react with BAL in the absence of NAD(+) The trimethylammonium group of BAL binds in the same position in the inactivating or productive modes. Accordingly, BAL does not inactivate the C(450)SSoBADH mutant and the degree of inactivation of the A(441)I and A(441)C mutants corresponds to their very different abilities to bind the trimethylammonium group. Cys(450)and the neighbouring residues that participate in stabilizing the thiohemiacetal are strictly conserved in plant ALDH10 enzymes with proven or predicted BADH activity, suggesting that inactivation by BAL is their common feature. Under osmotic stress conditions, this novel partial and reversible covalent regulatory mechanism may contribute to preventing NAD(+)exhaustion, while still permitting the synthesis of high amounts of GB and avoiding the accumulation of the toxic BAL.

  5. Irreversible APC(Cdh1) Inactivation Underlies the Point of No Return for Cell-Cycle Entry.

    PubMed

    Cappell, Steven D; Chung, Mingyu; Jaimovich, Ariel; Spencer, Sabrina L; Meyer, Tobias

    2016-06-30

    Proliferating cells must cross a point of no return before they replicate their DNA and divide. This commitment decision plays a fundamental role in cancer and degenerative diseases and has been proposed to be mediated by phosphorylation of retinoblastoma (Rb) protein. Here, we show that inactivation of the anaphase-promoting complex/cyclosome (APC(Cdh1)) has the necessary characteristics to be the point of no return for cell-cycle entry. Our study shows that APC(Cdh1) inactivation is a rapid, bistable switch initiated shortly before the start of DNA replication by cyclin E/Cdk2 and made irreversible by Emi1. Exposure to stress between Rb phosphorylation and APC(Cdh1) inactivation, but not after APC(Cdh1) inactivation, reverted cells to a mitogen-sensitive quiescent state, from which they can later re-enter the cell cycle. Thus, APC(Cdh1) inactivation is the commitment point when cells lose the ability to return to quiescence and decide to progress through the cell cycle. PMID:27368103

  6. Irreversible APC(Cdh1) Inactivation Underlies the Point of No Return for Cell-Cycle Entry.

    PubMed

    Cappell, Steven D; Chung, Mingyu; Jaimovich, Ariel; Spencer, Sabrina L; Meyer, Tobias

    2016-06-30

    Proliferating cells must cross a point of no return before they replicate their DNA and divide. This commitment decision plays a fundamental role in cancer and degenerative diseases and has been proposed to be mediated by phosphorylation of retinoblastoma (Rb) protein. Here, we show that inactivation of the anaphase-promoting complex/cyclosome (APC(Cdh1)) has the necessary characteristics to be the point of no return for cell-cycle entry. Our study shows that APC(Cdh1) inactivation is a rapid, bistable switch initiated shortly before the start of DNA replication by cyclin E/Cdk2 and made irreversible by Emi1. Exposure to stress between Rb phosphorylation and APC(Cdh1) inactivation, but not after APC(Cdh1) inactivation, reverted cells to a mitogen-sensitive quiescent state, from which they can later re-enter the cell cycle. Thus, APC(Cdh1) inactivation is the commitment point when cells lose the ability to return to quiescence and decide to progress through the cell cycle.

  7. Ca2+-induced activation and irreversible inactivation of chloride channels in the perfused plasmalemma of Nitellopsis obtusa.

    PubMed

    Kataev, A A; Zherelova, O M; Berestovsky, G N

    1984-12-01

    Experiments were carried out on the algal cells with removed tonoplast using both continuous intracellular perfusion and voltage clamp on plasmalemma. The transient plasmalemma current induced by depolarization disappeared upon perfusion with the Ca2+-chelating agent, EGTA, since the voltage-dependent calcium channels lost their ability to activate. Subsequent replacement of the perfusion medium containing EGTA by another with Ca2+ for clamped plasmalemma (-100 mV) induced an inward C1- current which showed both activation and inactivation. The maximal amplitude of the current at [C1-]in = 15 mmol/l (which is similar to that in native cells) was approximately twice that in electrically excited cell in vivo. The inactivation of C1 channels in the presence of internal Ca2+ was irreversible and had a time constant of 1-3 min. This supports our earlier suggestion (Lunevsky et al. 1983) that the inactivation of C1 channels in an intact cell (with a time constant of 1-3 s) is due to a decrease in Ca2+ concentration rather than to the activity of their own inactivation mechanism. The C1 channel selectivity sequence was following: C1- much greater than CH3SO-4 approximately equal to K+ much greater than SO2-4 (PK/PSO4 approximately 10). Activation of one half the channels occurs at a Ca2+ concentration of 2 X 10(-5) mol/l. Sr2+ also (though to a lesser extent) activated C1 channels but had to be present in a much more higher concentration than Ca2+. Mg2+ and Ba2+ appeared ineffective. Ca2+ activation did not, apparently, require participation of water-soluble intermediator including ATP. Thus, C1 channel functioning is controlled by Ca2+-, Sr2+-sensitive elements of the subplasmalemma cytoskeleton. PMID:6099298

  8. Irreversible UV inactivation of Cryptosporidium spp. despite the presence of UV repair genes.

    PubMed

    Rochelle, Paul A; Fallar, Daffodil; Marshall, Marilyn M; Montelone, Beth A; Upton, Steve J; Woods, Keith

    2004-01-01

    Ultraviolet light is being considered as a disinfectant by the water industry because it appears to be very effective for inactivating pathogens, including Cryptosporidium parvum. However, many organisms have mechanisms for repairing ultraviolet light-induced DNA damage, which may limit the utility of this disinfection technology. Inactivation of C. parvum was assessed by measuring infectivity in cells of the human ileocecal adenocarcinoma HCT-8 cell line, with an assay targeting a heat shock protein gene and using a reverse transcriptase polymerase chain reaction to detect infections. Oocysts of five different isolates displayed similar sensitivity to ultraviolet light. An average dosage of 7.6 mJ/cm2 resulted in 99.9% inactivation, providing the first evidence that multiple isolates of C. parvum are equally sensitive to ultraviolet disinfection. Irradiated oocysts were unable to regain pre-irradiation levels of infectivity, following exposure to a broad array of potential repair conditions, such as prolonged incubation, pre-infection excystation triggers, and post-ultraviolet holding periods. A combination of data-mining and sequencing was used to identify genes for all of the major components of a nucleotide excision repair complex in C. parvum and Cryptosporidium hominis. The average similarity between the two organisms for the various genes was 96.4% (range, 92-98%). Thus, while Cryptosporidum spp. may have the potential to repair ultraviolet light-induced damage, oocyst reactivation will not occur under the standard conditions used for storage and distribution of treated drinking water.

  9. Identification of SNAIL1 Peptide-Based Irreversible Lysine-Specific Demethylase 1-Selective Inactivators.

    PubMed

    Itoh, Yukihiro; Aihara, Keisuke; Mellini, Paolo; Tojo, Toshifumi; Ota, Yosuke; Tsumoto, Hiroki; Solomon, Viswas Raja; Zhan, Peng; Suzuki, Miki; Ogasawara, Daisuke; Shigenaga, Akira; Inokuma, Tsubasa; Nakagawa, Hidehiko; Miyata, Naoki; Mizukami, Tamio; Otaka, Akira; Suzuki, Takayoshi

    2016-02-25

    Inhibition of lysine-specific demethylase 1 (LSD1), a flavin-dependent histone demethylase, has recently emerged as a new strategy for treating cancer and other diseases. LSD1 interacts physically with SNAIL1, a member of the SNAIL/SCRATCH family of transcription factors. This study describes the discovery of SNAIL1 peptide-based inactivators of LSD1. We designed and prepared SNAIL1 peptides bearing a propargyl amine, hydrazine, or phenylcyclopropane moiety. Among them, peptide 3, bearing hydrazine, displayed the most potent LSD1-inhibitory activity in enzyme assays. Kinetic study and mass spectrometric analysis indicated that peptide 3 is a mechanism-based LSD1 inhibitor. Furthermore, peptides 37 and 38, which consist of cell-membrane-permeable oligoarginine conjugated with peptide 3, induced a dose-dependent increase of dimethylated Lys4 of histone H3 in HeLa cells, suggesting that they are likely to exhibit LSD1-inhibitory activity intracellularly. In addition, peptide 37 decreased the viability of HeLa cells. We believe this new approach for targeting LSD1 provides a basis for development of potent selective inhibitors and biological probes for LSD1. PMID:26700437

  10. Partial purification of the mu opioid receptor irreversibly labeled with (/sup 3/H)b-funaltrexamine

    SciTech Connect

    Liu-Chen, L.Y.; Phillips, C.A.; Tam, S.W.

    1986-03-01

    The mu opioid receptor in bovine striatal membranes was specifically and irreversibly labeled by incubation with 5 nM (/sup 3/H)..beta..-funaltrexamine (approx.-FNA) at 37/sup 0/C for 90 min in the presence of 100 mM NaCl. The specific and irreversible binding of (/sup 3/H)..beta..-FNA as defined by that blocked by 1 /sup +/M naloxone was about 60% of total irreversible binding. The specific irreversible binding was saturable, stereospecific, time-, temperature, and tissue-dependent. Mu opioid ligands were much more potent than delta or kappa ligands in inhibiting the specific irreversible labeling. SDS polyacrylamide gel electrophoresis of solubilized membranes in the presence of 2-mercaptoethanol yielded a major radiolabeled broad band of MW 68-97K daltons, characteristic of a glycoprotein band. This band was not observed in membranes labeled in the presence of excess unlabeled naloxone. The glycoprotein nature of the (/sup 3/H)..beta..-FNA-labeled opioid receptor was confirmed by its binding to a wheat germ agglutinin-Sepharose column and its elution with N-acetylglucosamine.

  11. Species Differences in the Oxidative Desulfurization of a Thiouracil-Based Irreversible Myeloperoxidase Inactivator by Flavin-Containing Monooxygenase Enzymes.

    PubMed

    Eng, Heather; Sharma, Raman; Wolford, Angela; Di, Li; Ruggeri, Roger B; Buckbinder, Leonard; Conn, Edward L; Dalvie, Deepak K; Kalgutkar, Amit S

    2016-08-01

    N1-Substituted-6-arylthiouracils, represented by compound 1 [6-(2,4-dimethoxyphenyl)-1-(2-hydroxyethyl)-2-thioxo-2,3-dihydropyrimidin-4(1H)-one], are a novel class of selective irreversible inhibitors of human myeloperoxidase. The present account is a summary of our in vitro studies on the facile oxidative desulfurization in compound 1 to a cyclic ether metabolite M1 [5-(2,4-dimethoxyphenyl)-2,3-dihydro-7H-oxazolo[3,2-a]pyrimidin-7-one] in NADPH-supplemented rats (t1/2 [half-life = mean ± S.D.] = 8.6 ± 0.4 minutes) and dog liver microsomes (t1/2 = 11.2 ± 0.4 minutes), but not in human liver microsomes (t1/2 > 120 minutes). The in vitro metabolic instability also manifested in moderate-to-high plasma clearances of the parent compound in rats and dogs with significant concentrations of M1 detected in circulation. Mild heat deactivation of liver microsomes or coincubation with the flavin-containing monooxygenase (FMO) inhibitor imipramine significantly diminished M1 formation. In contrast, oxidative metabolism of compound 1 to M1 was not inhibited by the pan cytochrome P450 inactivator 1-aminobenzotriazole. Incubations with recombinant FMO isoforms (FMO1, FMO3, and FMO5) revealed that FMO1 principally catalyzed the conversion of compound 1 to M1. FMO1 is not expressed in adult human liver, which rationalizes the species difference in oxidative desulfurization. Oxidation by FMO1 followed Michaelis-Menten kinetics with Michaelis-Menten constant, maximum rate of oxidative desulfurization, and intrinsic clearance values of 209 μM, 20.4 nmol/min/mg protein, and 82.7 μl/min/mg protein, respectively. Addition of excess glutathione essentially eliminated the conversion of compound 1 to M1 in NADPH-supplemented rat and dog liver microsomes, which suggests that the initial FMO1-mediated S-oxygenation of compound 1 yields a sulfenic acid intermediate capable of redox cycling to the parent compound in a glutathione-dependent fashion or undergoing further oxidation to a more

  12. Omi/HtrA2 catalytic cleavage of inhibitor of apoptosis (IAP) irreversibly inactivates IAPs and facilitates caspase activity in apoptosis.

    PubMed

    Yang, Qi-Heng; Church-Hajduk, Robin; Ren, Jinyu; Newton, Michelle L; Du, Chunying

    2003-06-15

    Omi/HtrA2 is a mitochondrial serine protease that is released into the cytosol during apoptosis to antagonize inhibitors of apoptosis (IAPs) and contribute to caspase-independent cell death. Here, we demonstrate that Omi/HtrA2 directly cleaves various IAPs in vitro, and the cleavage efficiency is determined by its IAP-binding motif, AVPS. Cleavage of IAPs such as c-IAP1 substantially reduces its ability to inhibit and ubiquitylate caspases. In contrast to the stoichiometric anti-IAP activity by Smac/DIABLO, Omi/HtrA2 cleavage of c-IAP1 is catalytic and irreversible, thereby more efficiently inactivating IAPs and promoting caspase activity. Elimination of endogenous Omi by RNA interference abolishes c-IAP1 cleavage and desensitizes cells to apoptosis induced by TRAIL. In addition, overexpression of cleavage-site mutant c-IAP1 makes cells more resistant to TRAIL-induced caspase activation. This IAP cleavage by Omi is independent of caspase. Taken together, these results indicate that unlike Smac/DIABLO, Omi/HtrA2's catalytic cleavage of IAPs is a key mechanism for it to irreversibly inactivate IAPs and promote apoptosis.

  13. On the involvement of intramolecular protein disulfide in the irreversible inactivation of 3-hydroxy-3-methylglutaryl-CoA reductase by diallyl disulfide.

    PubMed

    Omkumar, R V; Kadam, S M; Banerji, A; Ramasarma, T

    1993-06-24

    Treatment with diallyl disulfide, a constituent of garlic oil, irreversibly inactivated microsomal and a soluble 50 kDa form of HMG-CoA reductase. No radioactivity was found to be protein-bound on treating the soluble enzyme with [35S]diallyl disulfide, indicating the absence of the mixed disulfide of the type allyl-S-S-protein. SDS-PAGE and Western blot analyses of the diallyl-disulfide-treated protein showed no traces of the dimer of the type protein-S-S-protein, but clearly indicated BME-reversible increased mobility, as expected of an intramolecular protein disulfide. The sulfhydryl groups, as measured by alkylation with iodo[2-14C]acetic acid, were found to decrease in the diallyl-disulfide-treated enzyme protein. Tryptic peptide analysis also gave support for the possible presence of disulfide-containing peptides in such a protein. It appears that diallyl disulfide inactivated HMG-CoA reductase by forming an internal protein disulfide that became inaccessible for reduction by DTT, and thereby retaining the inactive state of the enzyme. PMID:8518292

  14. Active site directed irreversible inactivation of brewers' yeast pyruvate decarboxylase by the conjugated substrate analogue (E)-4-(4-chlorophenyl)-2-oxo-3-butenoic acid: development of a suicide substrate.

    PubMed

    Kuo, D J; Jordan, F

    1983-08-01

    (E)-4-(4-Chlorophenyl)-2-oxo-3-butenoic acid (CPB) was found to irreversibly inactivate brewers' yeast pyruvate decarboxylase (PDC, EC 4.1.1.1) in a biphasic, sigmoidal manner, as is found for the kinetic behavior of substrate. An expression was derived for two-site irreversible inhibition of allosteric enzymes, and the kinetic behavior of CPB fit the expression for two-site binding. The calculated Ki's of 0.7 mM and 0.3 mM for CPB were assigned to the catalytic site and the regulatory site, respectively. The presence of pyruvic acid at high concentrations protected PDC from inactivation, whereas low concentrations of pyruvic acid accelerated inactivation by CPB. Pyruvamide, a known allosteric activator of PDC, was found to enhance inactivation by CPB. The results can be explained if pyruvamide binds only to a regulatory site, but CPB and pyruvic acid compete for both the regulatory and the catalytic centers. [1-14C]CPB was found to lose 14CO2 concurrently with the inactivation of the enzyme. Therefore, CPB was being turned over by PDC, in addition to inactivating it. CPB can be labeled a suicide-type inactivator for PDC.

  15. Interactions of peptide triazole thiols with Env gp120 induce irreversible breakdown and inactivation of HIV-1 virions

    PubMed Central

    2013-01-01

    Background We examined the underlying mechanism of action of the peptide triazole thiol, KR13 that has been shown previously to specifically bind gp120, block cell receptor site interactions and potently inhibit HIV-1 infectivity. Results KR13, the sulfhydryl blocked KR13b and its parent non-sulfhydryl peptide triazole, HNG156, induced gp120 shedding but only KR13 induced p24 capsid protein release. The resulting virion post virolysis had an altered morphology, contained no gp120, but retained gp41 that bound to neutralizing gp41 antibodies. Remarkably, HIV-1 p24 release by KR13 was inhibited by enfuvirtide, which blocks formation of the gp41 6-helix bundle during membrane fusion, while no inhibition of p24 release occurred for enfuvirtide-resistant virus. KR13 thus appears to induce structural changes in gp41 normally associated with membrane fusion and cell entry. The HIV-1 p24 release induced by KR13 was observed in several clades of HIV-1 as well as in fully infectious HIV-1 virions. Conclusions The antiviral activity of KR13 and its ability to inactivate virions prior to target cell engagement suggest that peptide triazole thiols could be highly effective in inhibiting HIV transmission across mucosal barriers and provide a novel probe to understand biochemical signals within envelope that are involved in membrane fusion. PMID:24330857

  16. Quantum mechanics-based scoring rationalizes the irreversible inactivation of parasitic Schistosoma mansoni cysteine peptidase by vinyl sulfone inhibitors.

    PubMed

    Fanfrlík, Jindřich; Brahmkshatriya, Pathik S; Řezáč, Jan; Jílková, Adéla; Horn, Martin; Mareš, Michael; Hobza, Pavel; Lepšík, Martin

    2013-12-01

    The quantum mechanics (QM)-based scoring function that we previously developed for the description of noncovalent binding in protein-ligand complexes has been modified and extended to treat covalent binding of inhibitory ligands. The enhancements are (i) the description of the covalent bond breakage and formation using hybrid QM/semiempirical QM (QM/SQM) restrained optimizations and (ii) the addition of the new ΔG(cov)' term to the noncovalent score, describing the "free" energy difference between the covalent and noncovalent complexes. This enhanced QM-based scoring function is applied to a series of 20 vinyl sulfone-based inhibitory compounds inactivating the cysteine peptidase cathepsin B1 of the Schistosoma mansoni parasite (SmCB1). The available X-ray structure of the SmCB1 in complex with a potent vinyl sulfone inhibitor K11017 is used as a template to build the other covalently bound complexes and to model the derived noncovalent complexes. We present the correlation of the covalent score and its constituents with the experimental binding data. Four outliers are identified. They contain bulky R1' substituents structurally divergent from the template, which might induce larger protein rearrangements than could be accurately modeled. In summary, we propose a new computational approach and an optimal protocol for the rapid evaluation and prospective design of covalent inhibitors with a conserved binding mode. PMID:24195769

  17. Partial X chromosome trisomy with functional disomy of Xp due to failure of X inactivation

    SciTech Connect

    Gustashaw, K.M.; Zurcher, V.; Dickerman, L.H.; Stallard, R.; Willard, H.F.

    1994-10-15

    A 5-month-old girl with mild phenotypic abnormalities, developmental delay, and seizures was found to have the de novo karyotype 46,XX,-13,+der(13)t(X;13)(p21.2;p11.1). The partial trisomy of Xp21.2 {yields} pter was confirmed with fluorescence in situ hybridization, using an X chromosome painting probe and several cosmid and YAC probes for Xp sequences. Replication banding showed that one of the structurally normal X chromosomes was late-replicating, but that the Xp segment of the der(13) was early-replicating in all cells examined. Since segments of the X chromosome separated from the X inactivation center in Xq13.2 cannot undergo X inactivation, the result is functional disomy of distal Xp. As the loss of short arm material from chromosome 13 is not considered to be clinically significant, the genomic imbalance of Xp expressed in this patient most likely accounts for her abnormal phenotype. 39 refs., 5 figs., 1 tab.

  18. Pharmacokinetics and Disposition of the Thiouracil Derivative PF-06282999, an Orally Bioavailable, Irreversible Inactivator of Myeloperoxidase Enzyme, Across Animals and Humans.

    PubMed

    Dong, Jennifer Q; Varma, Manthena V; Wolford, Angela; Ryder, Tim; Di, Li; Feng, Bo; Terra, Steven G; Sagawa, Kazuko; Kalgutkar, Amit S

    2016-02-01

    The thiouracil derivative PF-06282999 [2-(6-(5-chloro-2-methoxyphenyl)-4-oxo-2-thioxo-3,4-dihydropyrimidin-1(2H)-yl)acetamide] is an irreversible inactivator of myeloperoxidase and is currently in clinical trials for the potential treatment of cardiovascular diseases. Concerns over idiosyncratic toxicity arising from bioactivation of the thiouracil motif to reactive species in the liver have been largely mitigated through the physicochemical (molecular weight, lipophilicity, and topological polar surface area) characteristics of PF-06282999, which generally favor elimination via nonmetabolic routes. To test this hypothesis, pharmacokinetics and disposition studies were initiated with PF-06282999 using animals and in vitro assays, with the ultimate goal of predicting human pharmacokinetics and elimination mechanisms. Consistent with its physicochemical properties, PF-06282999 was resistant to metabolic turnover from liver microsomes and hepatocytes from animals and humans and was devoid of cytochrome P450 inhibition. In vitro transport studies suggested moderate intestinal permeability and minimal transporter-mediated hepatobiliary disposition. PF-06282999 demonstrated moderate plasma protein binding across all of the species. Pharmacokinetics in preclinical species characterized by low to moderate plasma clearances, good oral bioavailability at 3- to 5-mg/kg doses, and renal clearance as the projected major clearance mechanism in humans. Human pharmacokinetic predictions using single-species scaling of dog and/or monkey pharmacokinetics were consistent with the parameters observed in the first-in-human study, conducted in healthy volunteers at a dose range of 20-200 mg PF-06282999. In summary, disposition characteristics of PF-06282999 were relatively similar across preclinical species and humans, with renal excretion of the unchanged parent emerging as the principal clearance mechanism in humans, which was anticipated based on its physicochemical properties and

  19. Partially irreversible paresis of the deep peroneal nerve caused by osteocartilaginous exostosis of the fibula without affecting the tibialis anterior muscle.

    PubMed

    Paprottka, Felix Julian; Machens, Hans-Günther; Lohmeyer, Jörn Andreas

    2012-08-01

    Dysfunction of the lower limb's muscles can cause severe impairment and immobilisation of the patient. As one of the leg's major motor and sensory nerves, the deep peroneal nerve (synonym: deep fibular nerve) plays a very important role in muscle innervation in the lower extremities. We report the case of a 19-year-old female patient, who suffered from a brace-like exostosis 6-cm underneath her left fibular head causing a partially irreversible paresis of her deep peroneal nerve. This nerve damage resulted in complete atrophy of her extensor digitorum longus and extensor hallucis longus muscle, and in painful sensory disturbance at her left shin and first web space. The tibialis anterior muscle stayed intact because its motor branch left the deep peroneal nerve proximal to the nerve lesion. Diagnosis was first verified 6 years after the onset of symptoms by a magnetic resonance imaging (MRI) scan of her complete left lower leg. Subsequently, the patient was operated on in our clinic, where a neurolysis was performed and the 4-cm-long osteocartilaginous exostosis was removed. Paralysis was already irreversible but sensibility returned completely after neurolysis. The presented case shows that an osteocartilaginous exostosis can be the cause for partial deep peroneal nerve paresis. If this disorder is diagnosed at an early stage, nerve damage is reversible. Typical for an exostosis is its first appearance during the juvenile growth phase.

  20. Partial inactivation of nucleus accumbens core decreases delay discounting in rats without affecting sensitivity to delay or magnitude.

    PubMed

    Moschak, Travis M; Mitchell, Suzanne H

    2014-07-15

    Increased preference for smaller, sooner rewards (delay discounting) is associated with several behavioral disorders, including ADHD and substance use disorders. However, delay discounting is a complex cognitive process and the relationship is unclear between the pathophysiology of the disorders and the component processes underlying delay discounting, including sensitivity to reinforcer delay and sensitivity to reinforcer magnitude. To investigate these processes, male Long Evans rats were trained in one of three tasks measuring sensitivity to delay, sensitivity to magnitude, or both (typical delay discounting task). After learning the task, animals were implanted with bilateral cannulae into either the nucleus accumbens core (AcbC) or the lateral orbitofrontal cortex (lOFC), both of which have been implicated in delay discounting. Upon recovering from the surgery, a baclofen/muscimol cocktail was infused to temporarily inactivate each of these two regions and task performance was assessed. Unlike previous studies showing that lesions of the AcbC increased delay discounting, partial inactivation of the AcbC decreased delay discounting, although it had no effects on the tasks independently assessing either sensitivity to delay or magnitude. The effects of AcbC inactivation were larger in animals that had low levels of delay discounting at baseline. Inactivation of the lOFC had no effects on behavior in any task. These findings suggest that the AcbC may act to promote impulsive choice in individuals with low impulsivity. Furthermore, the data suggest that the AcbC is able to modulate delay and magnitude sensitivity together, but not either of the two in isolation.

  1. X inactivation in Rett syndrome: A preliminary study showing partial preferential inactivation of paternal X with the M27{beta} probe

    SciTech Connect

    Camus, P.; Abbadi, N.; Gilgenkrantz, S.

    1994-04-15

    Rett syndrome (RS) is a severe progressive neurological disorder occurring exclusively in females. Most cases are sporadic. The few familial cases (less than 1%) cannot be explained by a simple mode of inheritance. Several hypotheses have been proposed: X-linked male lethal mutation, maternal uniparental disomy, fresh mutation on the X chromosome, involvement of mitochondrial DNA and differential inactivation with metabolic interference of X-borne alleles. The authors have examined the pattern of X inactivation in 10 affected girls who were selected according to the clinical criteria previously described and accepted by the French Rett Scientific Committee. The X inactivation pattern was studied by analysis of methylation at the hypervariable locus DXS255 with the M27{beta} probe. The results show a more-or-less skewed inactivation of paternal X in 8 Rett females, and 2 cases of symmetrical inactivation. In control girls, inactivation was symmetrical cases and the maternal X has been preferentially inactivated in the other 2 cases. In no case was a total skewed inactivation observed. Though there was clear evidence for a preferential paternal X inactivation that was statistically significant further studies are necessary to establish a relationship between X inactivation pattern and Rett syndrome.

  2. The inactivation of the sortilin gene leads to a partial disruption of prosaposin trafficking to the lysosomes

    SciTech Connect

    Zeng, Jibin; Racicott, Jesse; Morales, Carlos R.

    2009-11-01

    Lysosomes are intracellular organelles which contain enzymes and activator proteins involved in the digestion and recycling of a variety of cellular and extracellular substances. We have identified a novel sorting receptor, sortilin, which is involved in the lysosomal trafficking of the sphingolipid activator proteins, prosaposin and GM{sub 2}AP, and the soluble hydrolases cathepsin D, cathepsin H, and acid sphingomyelinase. Sortilin belongs to a growing family of receptors with homology to the yeast Vps10 protein, which acts as a lysosomal sorting receptor for carboxypeptidase Y. In this study we examined the effects of the sortilin gene inactivation in mice. The inactivation of this gene did not yield any noticeable lysosomal pathology. To determine the existence of an alternative receptor complementing the sorting function of sortilin, we quantified the concentration of prosaposin in the lysosomes of the nonciliated epithelial cells lining the efferent ducts. These cells were chosen because they express sortilin and have a large number of lysosomes containing prosaposin. In addition, the nonciliated cells are known to endocytose luminal prosaposin that is synthesized and secreted by Sertoli cells into the seminiferous luminal fluids. Consequently, the nonciliated cells are capable of targeting both exogenous and endogenous prosaposin to the lysosomes. Using electron microscope immunogold labeling and quantitative analysis, our results demonstrate that inactivation of the sortilin gene produces a significant decrease of prosaposin in the lysosomes. When luminal prosaposin was excluded from the efferent ducts, the level of prosaposin in lysosomes was even lower in the mutant mice. Nonetheless, a significant amount of prosaposin continues to reach the lysosomal compartment. These results strongly suggest the existence of an alternative receptor that complements the function of sortilin and explains the lack of lysosomal storage disorders in the sortilin

  3. Partially Neutralizing Potency against Emerging Genotype I Virus among Children Received Formalin-Inactivated Japanese Encephalitis Virus Vaccine

    PubMed Central

    Fan, Yi-Chin; Chen, Jo-Mei; Chiu, Hsien-Chung; Chen, Yi-Ying; Lin, Jen-Wei; Shih, Chen-Chang; Chen, Chih-Ming; Chang, Chao-Chin; Chang, Gwong-Jen J.; Chiou, Shyan-Song

    2012-01-01

    Background Genotype I (GI) Japanese encephalitis virus (JEV) that replaced GIII virus has become the dominant circulating virus in Asia. Currently, all registered live and inactivated JEV vaccines are derived from genotype III viruses. In Taiwan, the compulsory JEV vaccination policy recommends that children receives four doses of formalin-inactivated Nakayama (GIII) JEV vaccine. Methodology/Principal Findings To evaluate the influence of genotype replacement on the post-vaccination viral neutralizing ability by GIII and GI viruses, the small panel of vaccinated-children serum specimens was assembled, and the reciprocal 50% plaque-reduction neutralizing antibody titers (PRNT50) were measured against Nakayama vaccine strain, CJN GIII human brain isolate and TC2009-1 GI mosquito isolate. The seropositivity rate (PRNT50≥1∶10) and geometric mean titers (GMT) against the TC2009-1 virus were the lowest among the three viruses. The protective threshold against the CJN and TC2009-1 viruses could only be achieved when the GMT against Nakayama virus was ≥1∶20 or ≥1∶80, respectively. Using undiluted vaccinees' sera, the enhancement of JEV infection in K562 cells was observed in some low or non-neutralizing serum specimens. Conclusions/Significance Our preliminary study has shown that neutralizing antibodies, elicited by the mouse brain-derived and formalin-inactivated JEV Nakayama vaccine among a limited number of vaccinees, have reduced neutralizing capacity against circulating GI virus, but more detailed studies are needed to address the potential impact on the future vaccine policy. PMID:23029592

  4. Ribosome-inactivating proteins

    PubMed Central

    Walsh, Matthew J; Dodd, Jennifer E; Hautbergue, Guillaume M

    2013-01-01

    Ribosome-inactivating proteins (RIPs) were first isolated over a century ago and have been shown to be catalytic toxins that irreversibly inactivate protein synthesis. Elucidation of atomic structures and molecular mechanism has revealed these proteins to be a diverse group subdivided into two classes. RIPs have been shown to exhibit RNA N-glycosidase activity and depurinate the 28S rRNA of the eukaryotic 60S ribosomal subunit. In this review, we compare archetypal RIP family members with other potent toxins that abolish protein synthesis: the fungal ribotoxins which directly cleave the 28S rRNA and the newly discovered Burkholderia lethal factor 1 (BLF1). BLF1 presents additional challenges to the current classification system since, like the ribotoxins, it does not possess RNA N-glycosidase activity but does irreversibly inactivate ribosomes. We further discuss whether the RIP classification should be broadened to include toxins achieving irreversible ribosome inactivation with similar turnovers to RIPs, but through different enzymatic mechanisms. PMID:24071927

  5. Inactivation of Human Cytochrome P450 3A4 and 3A5 by Dronedarone and N-Desbutyl Dronedarone.

    PubMed

    Hong, Yanjun; Chia, Yvonne Mei Fen; Yeo, Ray Hng; Venkatesan, Gopalakrishnan; Koh, Siew Kwan; Chai, Christina Li Lin; Zhou, Lei; Kojodjojo, Pipin; Chan, Eric Chun Yong

    2016-01-01

    Dronedarone is an antiarrhythmic agent approved in 2009 for the treatment of atrial fibrillation. An in-house preliminary study demonstrated that dronedarone inhibits cytochrome P450 (CYP) 3A4 and 3A5 in a time-dependent manner. This study aimed to investigate the inactivation of CYP450 by dronedarone. We demonstrated for the first time that both dronedarone and its main metabolite N-desbutyl dronedarone (NDBD) inactivate CYP3A4 and CYP3A5 in a time-, concentration-, and NADPH-dependent manner. For the inactivation of CYP3A4, the inactivator concentration at the half-maximum rate of inactivation and inactivation rate constant at an infinite inactivator concentration are 0.87 µM and 0.039 minute(-1), respectively, for dronedarone, and 6.24 µM and 0.099 minute(-1), respectively, for NDBD. For CYP3A5 inactivation, the inactivator concentration at the half-maximum rate of inactivation and inactivation rate constant at an infinite inactivator concentration are 2.19 µM and 0.0056 minute(-1) for dronedarone and 5.45 µM and 0.056 minute(-1) for NDBD. The partition ratios for the inactivation of CYP3A4 and CYP3A5 by dronedarone are 51.1 and 32.2, and the partition ratios for the inactivation of CYP3A4 and CYP3A5 by NDBD are 35.3 and 36.6. Testosterone protected both CYP3A4 and CYP3A5 from inactivation by dronedarone and NDBD. Although the presence of Soret peak confirmed the formation of a quasi-irreversible metabolite-intermediate complex between dronedarone/NDBD and CYP3A4/CYP3A5, partial recovery of enzyme activity by potassium ferricyanide illuminated an alternative irreversible mechanism-based inactivation (MBI). MBI of CYP3A4 and CYP3A5 was further supported by the discovery of glutathione adducts derived from the quinone oxime intermediates of dronedarone and NDBD. In conclusion, dronedarone and NDBD inactivate CYP3A4 and CYP3A5 via unique dual mechanisms of MBI and formation of the metabolite-intermediate complex. Our novel findings contribute new knowledge for

  6. Irreversibility with quantum trajectories.

    PubMed

    Wisniacki, D A; Borondo, F; Benito, R M

    2005-10-01

    Irreversibility is an important issue for many quantum processes. Loschmidt echoes, originally introduced as a way to gauge sensitivity to perturbations in quantum mechanics, have turned out to be a useful tool for its investigation. Following the philosophy supporting this idea, and using quantum trajectories as defined in the causal interpretation of quantum mechanics due to Bohm, we introduce in this paper a more informative alternative measure for irreversibility. The method is applied to the Bunimovich stadium billiard, a paradigmatic example of chaotic system, that constitutes an excellent model for mesoscopic devices.

  7. Irreversible quantum baker map.

    PubMed

    Łoziński, Artur; Pakoński, Prot; Zyczkowski, Karol

    2002-12-01

    We propose a generalization of the model of classical baker map on the torus, in which the images of two parts of the phase space do overlap. This transformation is irreversible and cannot be quantized by means of a unitary Floquet operator. A corresponding quantum system is constructed as a completely positive map acting in the space of density matrices. We investigate spectral properties of this superoperator and their link with the increase of the entropy of initially pure states.

  8. Irreversible magnetic switch

    SciTech Connect

    Karnowsky, M.M.; Yost, F.G.

    1991-12-31

    This invention is comprised of an irreversible magnetic switch containing a ferromagnetic amorphous metal having a predetermined crystallization temperature in its inductor magnetic path. With the incorporation of such material, the magnetic properties after cooling from a high temperature excursion above its crystallization temperature are only a fraction of the original value. The difference is used to provide a safety feature in the magnetic switch.

  9. Inactivation of rabies virus by hydrogen peroxide.

    PubMed

    Abd-Elghaffar, Asmaa A; Ali, Amal E; Boseila, Abeer A; Amin, Magdy A

    2016-02-01

    Development of safe and protective vaccines against infectious pathogens remains a challenge. Inactivation of rabies virus is a critical step in the production of vaccines and other research reagents. Beta-propiolactone (βPL); the currently used inactivating agent for rabies virus is expensive and proved to be carcinogenic in animals. This study aimed to investigate the ability of hydrogen peroxide (H2O2) to irreversibly inactivate rabies virus without affecting its antigenicity and immunogenicity in pursuit of finding safe, effective and inexpensive alternative inactivating agents. H2O2 3% rapidly inactivated a Vero cell adapted fixed rabies virus strain designated as FRV/K within 2h of exposure without affecting its antigenicity or immunogenicity. No residual infectious virus was detected and the H2O2-inactivated vaccine proved to be safe and effective when compared with the same virus harvest inactivated with the classical inactivating agent βPL. Mice immunized with H2O2-inactivated rabies virus produced sufficient level of antibodies and were protected when challenged with lethal CVS virus. These findings reinforce the idea that H2O2 can replace βPL as inactivating agent for rabies virus to reduce time and cost of inactivation process. PMID:26731189

  10. Inactivation of rabies virus by hydrogen peroxide.

    PubMed

    Abd-Elghaffar, Asmaa A; Ali, Amal E; Boseila, Abeer A; Amin, Magdy A

    2016-02-01

    Development of safe and protective vaccines against infectious pathogens remains a challenge. Inactivation of rabies virus is a critical step in the production of vaccines and other research reagents. Beta-propiolactone (βPL); the currently used inactivating agent for rabies virus is expensive and proved to be carcinogenic in animals. This study aimed to investigate the ability of hydrogen peroxide (H2O2) to irreversibly inactivate rabies virus without affecting its antigenicity and immunogenicity in pursuit of finding safe, effective and inexpensive alternative inactivating agents. H2O2 3% rapidly inactivated a Vero cell adapted fixed rabies virus strain designated as FRV/K within 2h of exposure without affecting its antigenicity or immunogenicity. No residual infectious virus was detected and the H2O2-inactivated vaccine proved to be safe and effective when compared with the same virus harvest inactivated with the classical inactivating agent βPL. Mice immunized with H2O2-inactivated rabies virus produced sufficient level of antibodies and were protected when challenged with lethal CVS virus. These findings reinforce the idea that H2O2 can replace βPL as inactivating agent for rabies virus to reduce time and cost of inactivation process.

  11. Irreversible Simulated Tempering

    NASA Astrophysics Data System (ADS)

    Sakai, Yuji; Hukushima, Koji

    2016-10-01

    An extended ensemble Monte Carlo algorithm is proposed by introducing a violation of the detailed balance condition to the update scheme of the inverse temperature in simulated tempering. Our method, irreversible simulated tempering, is constructed based on the framework of the skew detailed balance condition. By applying this method to the ferromagnetic Ising model in two dimensions on a square lattice as a benchmark, the dynamical behavior of the inverse temperature and an autocorrelation function of the magnetization are studied numerically. It is found that the relaxation dynamics of the inverse temperature changes qualitatively from diffusive to ballistic by violating the detailed balance condition. Consequently, the autocorrelation time of magnetization is several times smaller than that for the conventional algorithm satisfying the detailed balance condition.

  12. Intrinsically irreversible heat engine

    DOEpatents

    Wheatley, John C.; Swift, Gregory W.; Migliori, Albert

    1984-01-01

    A class of heat engines based on an intrinsically irreversible heat transfer process is disclosed. In a typical embodiment the engine comprises a compressible fluid that is cyclically compressed and expanded while at the same time being driven in reciprocal motion by a positive displacement drive means. A second thermodynamic medium is maintained in imperfect thermal contact with the fluid and bears a broken thermodynamic symmetry with respect to the fluid. the second thermodynamic medium is a structure adapted to have a low fluid flow impedance with respect to the compressible fluid, and which is further adapted to be in only moderate thermal contact with the fluid. In operation, thermal energy is pumped along the second medium due to a phase lag between the cyclical heating and cooling of the fluid and the resulting heat conduction between the fluid and the medium. In a preferred embodiment the engine comprises an acoustical drive and a housing containing a gas which is driven at a resonant frequency so as to be maintained in a standing wave. Operation of the engine at acoustic frequencies improves the power density and coefficient of performance. The second thermodynamic medium can be coupled to suitable heat exchangers to utilize the engine as a simple refrigeration device having no mechanical moving parts. Alternatively, the engine is reversible in function so as to be utilizable as a prime mover by coupling it to suitable sources and sinks of heat.

  13. Intrinsically irreversible heat engine

    DOEpatents

    Wheatley, J.C.; Swift, G.W.; Migliori, A.

    1984-12-25

    A class of heat engines based on an intrinsically irreversible heat transfer process is disclosed. In a typical embodiment the engine comprises a compressible fluid that is cyclically compressed and expanded while at the same time being driven in reciprocal motion by a positive displacement drive means. A second thermodynamic medium is maintained in imperfect thermal contact with the fluid and bears a broken thermodynamic symmetry with respect to the fluid. The second thermodynamic medium is a structure adapted to have a low fluid flow impedance with respect to the compressible fluid, and which is further adapted to be in only moderate thermal contact with the fluid. In operation, thermal energy is pumped along the second medium due to a phase lag between the cyclical heating and cooling of the fluid and the resulting heat conduction between the fluid and the medium. In a preferred embodiment the engine comprises an acoustical drive and a housing containing a gas which is driven at a resonant frequency so as to be maintained in a standing wave. Operation of the engine at acoustic frequencies improves the power density and coefficient of performance. The second thermodynamic medium can be coupled to suitable heat exchangers to utilize the engine as a simple refrigeration device having no mechanical moving parts. Alternatively, the engine is reversible in function so as to be utilizable as a prime mover by coupling it to suitable sources and sinks of heat. 11 figs.

  14. Intrinsically irreversible heat engine

    DOEpatents

    Wheatley, J.C.; Swift, G.W.; Migliori, A.

    1984-01-01

    A class of heat engines based on an intrinsically irreversible heat transfer process is disclosed. In a typical embodiment the engine comprises a compressible fluid that is cyclically compressed and expanded while at the same time being driven in reciprocal motion by a positive displacement drive means. A second thermodynamic medium is maintained in imperfect thermal contact with the fluid and bears a broken thermodynamic symmetry with respect to the fluid. The second thermodynamic medium is a structure adapted to have a low fluid flow impedance with respect to the compressible fluid, and which is further adapted to be in only moderate thermal contact with the fluid. In operation, thermal energy is pumped along the second medium due to a phase lag between the cyclical heating and cooling of the fluid and the resulting heat conduction between the fluid and the medium. In a preferred embodiment the engine comprises an acoustical drive and a housing containing a gas which is driven at a resonant frequency so as to be maintained in a standing wave. Operation of the engine at acoustic frequencies improves the power density and coefficient of performance. The second thermodynamic medium can be coupled to suitable heat exchangers to utilize the engine as a simple refrigeration device having no mechanical moving parts. Alternatively, the engine is reversible in function so as to be utilizable as a prime mover by coupling it to suitable sources and sinks of heat.

  15. Cyanide inactivation of hydrogenase from Azotobacter vinelandii

    SciTech Connect

    Seefeldt, L.C.; Arp, D.J. )

    1989-06-01

    The effects of cyanide on membrane-associated and purified hydrogenase from Azotobacter vinelandii were characterized. Inactivation of hydrogenase by cyanide was dependent on the activity (oxidation) state of the enzyme. Active (reduced) hydrogenase showed no inactivation when treated with cyanide over several hours. Treatment of reversibly inactive (oxidized) states of both membrane-associated and purified hydrogenase, however, resulted in a time-dependent, irreversible loss of hydrogenase activity. The rate of cyanide inactivation was dependent on the cyanide concentration and was an apparent first-order process for purified enzyme (bimolecular rate constant, 23.1 M{sup {minus}1} min{sup {minus}1} for CN{sup {minus}}). The rate of inactivation decreased with decreasing pH. ({sup 14}C)cyanide remained associated with cyanide-inactivated hydrogenase after gel filtration chromatography, with a stoichiometry of 1.7 mol of cyanide bound per mol of inactive enzyme. The presence of saturating concentrations of CO had no effect on the rate or extent of cyanide inactivation of hydrogenases. The results indicate that cyanide can cause a time-dependent, irreversible inactivation of hydrogenase in the oxidized, activatable state but has no effect when hydrogenase is in the reduced, active state.

  16. Irreversible thermal denaturation of Torpedo californica acetylcholinesterase.

    PubMed Central

    Kreimer, D. I.; Shnyrov, V. L.; Villar, E.; Silman, I.; Weiner, L.

    1995-01-01

    Thermal denaturation of Torpedo californica acetylcholinesterase, a disulfide-linked homodimer with 537 amino acids in each subunit, was studied by differential scanning calorimetry. It displays a single calorimetric peak that is completely irreversible, the shape and temperature maximum depending on the scan rate. Thus, thermal denaturation of acetylcholinesterase is an irreversible process, under kinetic control, which is described well by the two-state kinetic scheme N-->D, with activation energy 131 +/- 8 kcal/mol. Analysis of the kinetics of denaturation in the thermal transition temperature range, by monitoring loss of enzymic activity, yields activation energy of 121 +/- 20 kcal/mol, similar to the value obtained by differential scanning calorimetry. Thermally denatured acetylcholinesterase displays spectroscopic characteristics typical of a molten globule state, similar to those of partially unfolded enzyme obtained by modification with thiol-specific reagents. Evidence is presented that the partially unfolded states produced by the two different treatments are thermodynamically favored relative to the native state. PMID:8563632

  17. Oxidative inactivation of glutamine synthetase from the cyanobacterium Anabaena variabilis.

    PubMed Central

    Martin, G; Haehnel, W; Böger, P

    1997-01-01

    In crude extracts of the cyanobacterium Anabaena variabilis, glutamine synthetase (GS) could be effectively inactivated by the addition of NADH. GS inactivation was completed within 30 min. Both the inactivated GS and the active enzyme were isolated. No difference between the two enzyme forms was seen in sodium dodecyl sulfate-gels, and only minor differences were detectable by UV spectra, which excludes modification by a nucleotide. Mass spectrometry revealed that the molecular masses of active and inactive GS are equal. While the Km values of the substrates were unchanged, the Vmax values of the inactive GS were lower, reflecting the inactivation factor in the crude extract. This result indicates that the active site was affected. From the crude extract, a fraction mediating GS inactivation could be enriched by ammonium sulfate precipitation and gel filtration. GS inactivation by this fraction required the presence of NAD(P)H, Fe3+, and oxygen. In the absence of the GS-inactivating fraction, GS could be inactivated by Fe2+ and H2O2. The GS-inactivating fraction produced Fe2+ and H2O2, using NADPH, Fe3+, and oxygen. Accordingly, the inactivating fraction was inhibited by catalase and EDTA. This GS-inactivating system of Anabaena is similar to that described for oxidative GS inactivation in Escherichia coli. We conclude that GS inactivation by NAD(P)H is caused by irreversible oxidative damage and is not due to a regulatory mechanism of nitrogen assimilation. PMID:9006027

  18. [Thermal inactivation of alpha-galactosidase from Penicillium canescens].

    PubMed

    Borzova, N V; Varbanets, L D

    2010-01-01

    The kinetics and mechanism of thermal inactivation of Penicillium canescens alpha-galactosidase in the temperature range of 55-65 degrees C have been studied. The kinetic scheme of alpha-galactosidase thermal inactivation was proposed which included the reversible dissociation of active hexamers into associating monomers and irreversible denaturation of monomers. The kinetic constants of thermal inactivation have been determined. The effect of enzyme concentration and purification efficiency has been investigated. A possibility of defence of protein molecule from thermal inactivation in the presence of BSA, glycerol, melibiose, raffinose and stachyose is shown.

  19. Differential inactivation of alfalfa nodule glutamine synthetases by tabtoxinine-. beta. -lactam. [Pseudomonas syringae

    SciTech Connect

    Knight, T.J.; Unkefer, P.J.

    1987-04-01

    The presence of the pathogen Pseudomonas syringae pv. tabaci within the rhizosphere of nodulated alfalfa plants results in an increase in N/sub 2/-fixation potential and growth, but a 40-50% decrease in nodule glutamine synthetase (GS) activity, as compared to nodulated control plants. Tabtoxinine-..beta..-Lactam an exocellular toxin produced by Pseudomonas syringae pv tabaci irreversibly inhibits glutamine synthetase. Partial purification of nodule GS by DEAE-cellulose chromatography reveals two enzyme forms are present (GS/sub n1/ and GS/sub n2/). In vitro inactivation of the two glutamine synthetases associated with the nodule indicates a differential sensitivity to T-..beta..-L. The nodule specific GS/sub n1/ is much less sensitive to T-..beta..-L than the GS/sub n2/ enzyme, which was found to coelute with the root enzyme (GS/sub r/). However, both GS/sub n1/ and GS/sub n2/ are rapidly inactivated by methionine sulfoximine, another irreversible inhibitor of GS.

  20. Kinetics of irreversible inhibition of yeast alcohol dehydrogenase during modification by o-phthaldehyde.

    PubMed

    Le, W P; Yan, S X; Huang, M Q; Zhang, Y X; Zhou, H M

    The kinetic theory of the substrate reaction during irreversible inhibition of enzyme activity described previously has been applied to a study on the kinetics of the course of inactivation of yeast alcohol dehydrogenase (YADH) by o-phthaldehyde (OPTA). The microscopic constants for the reaction of the inactivators with the free enzyme and with the enzyme-substrate complexes were determined. The inactivation is a monophasic pseudo-first-order reaction with OPTA. The apparent rate constant A is independent of the OPTA concentration, indicating that the inactivation is a noncomplexing inhibition. The marked protective effect of substrates on the inactivation of YADH by OPTA has been observed. This result suggests that the modification of the enzyme by OPTA may occur at the active site.

  1. Information symmetries in irreversible processes

    NASA Astrophysics Data System (ADS)

    Ellison, Christopher J.; Mahoney, John R.; James, Ryan G.; Crutchfield, James P.; Reichardt, Jörg

    2011-09-01

    We study dynamical reversibility in stationary stochastic processes from an information-theoretic perspective. Extending earlier work on the reversibility of Markov chains, we focus on finitary processes with arbitrarily long conditional correlations. In particular, we examine stationary processes represented or generated by edge-emitting, finite-state hidden Markov models. Surprisingly, we find pervasive temporal asymmetries in the statistics of such stationary processes. As a consequence, the computational resources necessary to generate a process in the forward and reverse temporal directions are generally not the same. In fact, an exhaustive survey indicates that most stationary processes are irreversible. We study the ensuing relations between model topology in different representations, the process's statistical properties, and its reversibility in detail. A process's temporal asymmetry is efficiently captured using two canonical unifilar representations of the generating model, the forward-time and reverse-time ɛ-machines. We analyze example irreversible processes whose ɛ-machine representations change size under time reversal, including one which has a finite number of recurrent causal states in one direction, but an infinite number in the opposite. From the forward-time and reverse-time ɛ-machines, we are able to construct a symmetrized, but nonunifilar, generator of a process—the bidirectional machine. Using the bidirectional machine, we show how to directly calculate a process's fundamental information properties, many of which are otherwise only poorly approximated via process samples. The tools we introduce and the insights we offer provide a better understanding of the many facets of reversibility and irreversibility in stochastic processes.

  2. Niche Applications of Irreversible Electroporation.

    PubMed

    Bhatia, Shivank S; Arya, Rahul; Narayanan, Govindarajan

    2015-09-01

    Irreversible electroporation (IRE) induces cell death by exposing it to high-voltage, low-energy DC current pulses. The mechanism of cell death and healing is a departure from the other existing technologies such as radiofrequency ablation, microwave ablation, and cryoablation. These thermal ablative technologies have several applications in oncology but have limitations that have also been established. IRE has shown promise to overcome some of these limitations and has enabled the use of an ablative technology in treating lesions close to the bile ducts and vasculature and in organs such as the pancreas. This review highlights some of the niche applications of IRE and the data so far.

  3. Irreversible gettering of thionyl chloride

    SciTech Connect

    LeRoy Whinnery; Steve Goods; George Buffleben; Tim Sheppodd

    1999-11-01

    The authors have successfully demonstrated the irreversible gettering of SOCl{sub 2} by ZnO/ASZMTEDA carbon over a modest temperature range. While thionyl chloride decomposition was slow below {minus}20 C, lower temperatures are expected to be less of a problem than at higher temperatures. The approximately 30 cc of thionyl chloride in a typical D-cell would require 50 g of ZnO and 107 g of ASZMTEDA carbon. Fortunately, since it is unlikely to happen at all, it is common practice to assume only one cell will fail (leak) in a given battery pack. So, one charge of getter can protect the whole battery pack. In summary, ZnO/ASZMTEDA carbon fulfills all of the requirements of an ideal getter including: irreversible binding or reaction with SOCl{sub 2}, high volumetric uptake capacity, high efficiency, non-volatile, air stable, insensitive to poisoning, non-toxic, cheap, non-corrosive, and the gettering product is not a liquid or oil that could block further flow or accessibility. Future work in this area includes incorporation of the ZnO and carbon into a structural open-celled porous monolith, as well as, gettering for other types of batteries (e.g., Li/MnO{sub 2}).

  4. Lyapunov decay in quantum irreversibility.

    PubMed

    García-Mata, Ignacio; Roncaglia, Augusto J; Wisniacki, Diego A

    2016-06-13

    The Loschmidt echo--also known as fidelity--is a very useful tool to study irreversibility in quantum mechanics due to perturbations or imperfections. Many different regimes, as a function of time and strength of the perturbation, have been identified. For chaotic systems, there is a range of perturbation strengths where the decay of the Loschmidt echo is perturbation independent, and given by the classical Lyapunov exponent. But observation of the Lyapunov decay depends strongly on the type of initial state upon which an average is carried out. This dependence can be removed by averaging the fidelity over the Haar measure, and the Lyapunov regime is recovered, as has been shown for quantum maps. In this work, we introduce an analogous quantity for systems with infinite dimensional Hilbert space, in particular the quantum stadium billiard, and we show clearly the universality of the Lyapunov regime. PMID:27140966

  5. [Circulatory survival of irreversible comas].

    PubMed

    Cartier, F; Chevet, D; Garré, M; Launois, B; Thomas, R; Le Pollès, R

    1975-01-18

    On the basis of a series of 53 cases of irreversible coma maintained in circulatory survival with the aim of removing the kidneys, the authors discuss the mode of treatment, with particular reference to the intravenous fluids used and the use of medications influencing the circulation. Fluid and electrolytes given must be adjusted hourly to ensure the exact replacement of urinary losses. Isoprotenerol is the only medication usually necessary. In the event of circulatory insufficiency, which is difficult to foresee and hence prevent, immediate volume expansion in a short a time as possible and isoprotenerol most frequently correct the situation (14 out of 17 cases). Thus effective circulation may be maintained until the kidneys are removed (48 out of 53 cases). 92 p.cent of the grafted kidneys functioned from the first day onwards. PMID:1093120

  6. Variational principles of irreversible processes

    NASA Astrophysics Data System (ADS)

    Ichiyanagi, Masakazu

    1994-07-01

    This article reviews developments of variational principles in the study of irreversible processes during the past three decades or so. The variational principles we consider here are related to entropy production. The purpose of this article is to explicate that we can formulate a variational principle which relates the transport coefficients to microscopic dynamics of fluctuations. The quantum variational principle restricts the nonequilibrium density matrix to a class conforming to the requirement demanded by the second law of thermodynamics. These are various kinds of variational principles according to different stages of a macroscopic system. The three stages are known, which are dynamical, kinetic, and thermodynamical stages. The relationships among these variational principles are discussed from the point of view of the contraction of information about irrelevant components. Nakano's variational principle has close similarity to the Lippmann-Schwinger theory of scattering, in which some incoming and outgoing disturbances have to be considered in a pair. It is also shown that the variational principle of Onsager's type can be reformulated in the form of Hamilton's principle if a generalization of Hamilton's principle proposed by Djukic and Vujanovic is used. A variational principle in the diagrammatic method is also reviewed, which utilizes the generalized Ward-Takahashi relations.

  7. Inactivation of Caliciviruses

    PubMed Central

    Nims, Raymond; Plavsic, Mark

    2013-01-01

    The Caliciviridae family of viruses contains clinically important human and animal pathogens, as well as vesivirus 2117, a known contaminant of biopharmaceutical manufacturing processes employing Chinese hamster cells. An extensive literature exists for inactivation of various animal caliciviruses, especially feline calicivirus and murine norovirus. The caliciviruses are susceptible to wet heat inactivation at temperatures in excess of 60 °C with contact times of 30 min or greater, to UV-C inactivation at fluence ≥30 mJ/cm2, to high pressure processing >200 MPa for >5 min at 4 °C, and to certain photodynamic inactivation approaches. The enteric caliciviruses (e.g.; noroviruses) display resistance to inactivation by low pH, while the non-enteric species (e.g.; feline calicivirus) are much more susceptible. The caliciviruses are inactivated by a variety of chemicals, including alcohols, oxidizing agents, aldehydes, and β-propiolactone. As with inactivation of viruses in general, inactivation of caliciviruses by the various approaches may be matrix-, temperature-, and/or contact time-dependent. The susceptibilities of the caliciviruses to the various physical and chemical inactivation approaches are generally similar to those displayed by other small, non-enveloped viruses, with the exception that the parvoviruses and circoviruses may require higher temperatures for inactivation, while these families appear to be more susceptible to UV-C inactivation than are the caliciviruses. PMID:24276023

  8. Ice-induced partial unfolding and aggregation of an integral membrane protein.

    PubMed

    Garber Cohen, Iona P; Castello, Pablo R; González Flecha, F Luis

    2010-11-01

    Although the deleterious effects of ice on water-soluble proteins are well established, little is known about the freeze stability of membrane proteins. Here we explore this issue through a combined kinetic and spectroscopic approach using micellar-purified plasma membrane calcium pump as a model. The ATPase activity of this protein significantly diminished after freezing using a slow-cooling procedure, with the decrease in the activity being an exponential function of the storage time at 253K, with t(½)=3.9±0.6h. On the contrary, no significant changes on enzyme activity were detected when a fast cooling procedure was performed. Regardless of the cooling rate, successive freeze-thaw cycles produced an exponential decrease in the Ca(2+)-ATPase activity, with the number of cycles at which the activity was reduced to half being 9.2±0.3 (fast cooling) and 3.7±0.2 (slow cooling). PAGE analysis showed that neither degradation nor formation of SDS-stable aggregates of the protein takes place during protein inactivation. Instead, the inactivation process was found to be associated with the irreversible partial unfolding of the polypeptide chain, as assessed by Trp fluorescence, far UV circular dichroism, and 1-anilino-naphtalene-8-sulfonate binding. This inactive protein undergoes, in a later stage, a further irreversible transformation leading to large aggregates.

  9. Blueberry polyphenol oxidase: Characterization and the kinetics of thermal and high pressure activation and inactivation.

    PubMed

    Terefe, Netsanet Shiferaw; Delon, Antoine; Buckow, Roman; Versteeg, Cornelis

    2015-12-01

    Partially purified blueberry polyphenol oxidase (PPO) in Mcllvaine buffer (pH=3.6, typical pH of blueberry juice) was subjected to processing at isothermal-isobaric conditions at temperatures from 30 to 80 °C and pressure from 0.1 to 700 MPa. High pressure processing at 30-50 °C at all pressures studied caused irreversible PPO activity increase with a maximum of 6.1 fold increase at 500 MPa and 30 °C. Treatments at mild pressure-mild temperature conditions (0.1-400 MPa, 60 °C) also caused up to 3 fold PPO activity increase. Initial activity increase followed by a decrease occurred at relatively high pressure-mild temperature (400-600 MPa, 60 °C) and mild pressure-high temperature (0.1-400 MPa, 70-80 °C) combinations. At temperatures higher than 76 °C, monotonic decrease in PPO activity occurred at 0.1 MPa and pressures higher than 500 MPa. The activation/inactivation kinetics of the enzyme was successfully modelled assuming consecutive reactions in series with activation followed by inactivation.

  10. Irritation of ocular tissue by irreversible hydrocolloids.

    PubMed

    Moergeli, J R; Fraleigh, E M; Ostrowski, J S; Pelleu, G B

    1985-08-01

    Two ophthalmic and two dental irreversible hydrocolloid materials were tested on rabbit conjunctivae to determine histologically their potential to irritate these tissues. Each of the four impression materials elicited nearly the same amount of inflammatory response. The differences between the response of the controls and the response to Ophthalmic Mold-Eye, Jelset Special Formula, and Kerr Alignate Type II were significant. These results indicate that certain dental irreversible hydrocolloids may be used for ocular prostheses but that they should be used with caution because of the inflammation caused by irreversible hydrocolloids.

  11. Factors in the inactivation of Encephalomyocarditis virus in aerosols.

    PubMed

    de Jong, J C; Harmsen, M; Trouwborst, T

    1975-07-01

    Encephalomyocarditis virus in aerosols is inactivated rapidly at relative humidities below 50%. In glycerol-water mixtures a similar decrease of infectivity occurs when the glycerol concentration exceeds 78% (wt/wt), corresponding to a relative humidity of 50%. The decay in aerosols does not involve oxygen or surface-dependent factors. Variation of temperature shows the inactivation to be a low-energy process with an activation enthalpy of 15 kcal per mol. The damage could be ascribed to dehydration of the virion, presumably proceeding to removal of structurally essential water molecules. This might trigger irreversible changes in the protein coat, resulting in disintegration of the virion.

  12. Nitric oxide and nitric oxide-generating agents induce a reversible inactivation of protein kinase C activity and phorbol ester binding.

    PubMed

    Gopalakrishna, R; Chen, Z H; Gundimeda, U

    1993-12-25

    Since S-nitrosylation of protein thiols is one of the cellular regulatory mechanisms induced by nitric oxide (NO), and since protein kinase C (PKC) has critical thiol residues which influence its kinase activity, we have determined whether NO could regulate this enzyme. Initial studies were carried out with purified PKC and the NO-generating agent S-nitrosocysteine. This agent decreased phosphotransferase activity of PKC in a Ca(2+)- and oxygen-dependent manner with an IC50 of 75 microM. Phorbol ester binding was affected partially only at higher concentrations (> 100 microM) of S-nitrosocysteine. This inactivation of PKC was blocked by the NO scavenger oxyhemoglobin or reversed by dithiothreitol. It is likely that NO initially induced an S-nitrosylation of vicinal thiols, which were then oxidized to form an intramolecular disulfide. Other NO-generating agents such as S-nitroso-N-acetylpenicillamine and sodium nitroprusside, as well as authentic NO gas, induced similar types of PKC modifications. In intact B16 melanoma cells treated with S-nitrosocysteine a rapid decrease in PKC activity in both cytosol and membrane was observed. Unlike in experiments with purified PKC, in intact cells treated with S-nitrosocysteine the phorbol ester binding also decreased to a rate equal to that of PKC activity. These modifications were readily reversed by treating the homogenates with dithiothreitol in test tubes or by removing the NO-generating source from intact cells. To determine whether the limited amounts of NO generated within the intact cells could induce this type of PKC modification, the macrophage cell line IC-21 was treated with lipopolysacharide and Ca2+ ionophore A23187 to induce the NO production. With an increase in generation of NO (3-12-h period) in these cells, a parallel and irreversible decrease in PKC activity and phorbol ester binding was observed. A specific inhibitor for NO synthase, NG-monomethyl-L-arginine, inhibited both the production of NO and PKC

  13. Irreversible thermodynamics of Poisson processes with reaction.

    PubMed

    Méndez, V; Fort, J

    1999-11-01

    A kinetic model is derived to study the successive movements of particles, described by a Poisson process, as well as their generation. The irreversible thermodynamics of this system is also studied from the kinetic model. This makes it possible to evaluate the differences between thermodynamical quantities computed exactly and up to second-order. Such differences determine the range of validity of the second-order approximation to extended irreversible thermodynamics.

  14. Irreversible thermodynamics of Poisson processes with reaction

    NASA Astrophysics Data System (ADS)

    Méndez, Vicenç; Fort, Joaquim

    1999-11-01

    A kinetic model is derived to study the successive movements of particles, described by a Poisson process, as well as their generation. The irreversible thermodynamics of this system is also studied from the kinetic model. This makes it possible to evaluate the differences between thermodynamical quantities computed exactly and up to second-order. Such differences determine the range of validity of the second-order approximation to extended irreversible thermodynamics.

  15. Heat devices in nonlinear irreversible thermodynamics

    NASA Astrophysics Data System (ADS)

    Izumida, Y.; Okuda, K.; Roco, J. M. M.; Hernández, A. Calvo

    2015-05-01

    We present results obtained by using nonlinear irreversible models for heat devices. In particular, we focus on the global performance characteristics, the maximum efficiency and the efficiency at maximum power regimes for heat engines, and the maximum coefficient of performance (COP) and the COP at maximum cooling power regimes for refrigerators. We analyze the key role played by the interplay between irreversibilities coming from heat leaks and internal dissipations. We also discuss the relationship between these results and those obtained by different models.

  16. Studies on the mechanism of activation and inactivation of pyruvate,phosphate dikinase. A possible regulatory role for the enzyme in the C4 dicarboxylic acid pathway of photosynthesis

    PubMed Central

    Hatch, M. D.; Slack, C. R.

    1969-01-01

    1. The activity of pyruvate,Pi dikinase in leaves of maize and Amaranthus palmeri rapidly falls on transferring illuminated plants to darkness. Illumination of dark-treated plants results in an immediate rapid increase in activity of the enzyme, the final activity reached being dependent on the intensity of the incident light. 2. Activation of the enzyme in extracts of dark-treated maize leaves after gel filtration on Sephadex G-25 requires a thiol and Pi. The Pi requirement for activation can be replaced by arsenate. Activation of the enzyme is inhibited by AMP and GMP and possibly also by ADP and ATP. Activation of the enzyme after gel filtration on Sephadex G-200 also requires a heat-labile component that is excluded by Sephadex G-25. 3. The active enzyme isolated from illuminated leaves is inactivated by ADP in the presence of a thiol, the rate of inactivation being very much faster in air than in an oxygen-free atmosphere. Reactivation of the ADP-inactivated enzyme requires a thiol, Pi and a component excluded by Sephadex G-25 but considerably retarded by Sephadex G-200. 4. The active enzyme is rapidly and irreversibly inactivated in the absence of a thiol. Inactivation is accelerated by both sodium diethyldithiocarbamate and tetraethylthiuram disulphide, and the enzyme inactivated by these reagents is completely reactivated by incubation with dithiothreitol. This reactivation does not require Pi. The inactive enzyme from dark-treated leaves is stabilized by diethyldithiocarbamate and can be partially activated by dithiothreitol alone; complete reactivation requires both dithiothreitol and Pi. 5. The enzyme activity is markedly inhibited by the thiol reagents p-chloromercuribenzoate, γ-(p-arsenophenyl)-n-butyrate and an equimolar mixture of arsenite and 2,3-dimercaptopropan-1-ol. 6. The processes of activation and inactivation observed in vitro are discussed in relation to the regulation of pyruvate,Pi dikinase activity in the leaf. PMID:5821721

  17. [Irreversible image compression in radiology. Current status].

    PubMed

    Pinto dos Santos, D; Jungmann, F; Friese, C; Düber, C; Mildenberger, P

    2013-03-01

    Due to increasing amounts of data in radiology methods for image compression appear both economically and technically interesting. Irreversible image compression allows markedly higher reduction of data volume in comparison with reversible compression algorithms but is, however, accompanied by a certain amount of mathematical and visual loss of information. Various national and international radiological societies have published recommendations for the use of irreversible image compression. The degree of acceptable compression varies across modalities and regions of interest.The DICOM standard supports JPEG, which achieves compression through tiling, DCT/DWT and quantization. Although mathematical loss due to rounding up errors and reduction of high frequency information occurs this results in relatively low visual degradation.It is still unclear where to implement irreversible compression in the radiological workflow as only few studies analyzed the impact of irreversible compression on specialized image postprocessing. As long as this is within the limits recommended by the German Radiological Society irreversible image compression could be implemented directly at the imaging modality as it would comply with § 28 of the roentgen act (RöV). PMID:23456043

  18. N-Alkoxyheterocycles As Irreversible Photooxidants†

    PubMed Central

    Wosinska, Zofia M.; Stump, Faye L.; Ranjan, Rajeev; Lorance, Edward D.; Finley, GeNita N.; Patel, Priya P.; Khawaja, Muzamil A.; Odom, Katie L.; Kramer, Wolfgang H.; Gould, Ian R.

    2015-01-01

    Irreversible photooxidation based on N–O bond fragmentation is demonstrated for N-methoxyheterocycles in both the singlet and triplet excited state manifolds. The energetic requirements for bond fragmentation are studied in detail. Bond fragmentation in the excited singlet manifold is possible for ππ* singlet states with energies significantly larger than the N–O bond dissociation energy of ca 55 kcal mol−1. For the nπ* triplet states, N–O bond fragmentation does not occur in the excited state for orbital overlap and energetic reasons. Irreversible photooxidation occurs in the singlet states by bond fragmentation followed by electron transfer. Irreversible photooxidation occurs in the triplet states via bimolecular electron transfer to the donor followed by bond fragmentation. Using these two sensitization schemes, donors can be irreversibly oxidized with oxidation potentials ranging from ca 1.6–2.2 V vs SCE. The corresponding N-ethylheterocycles are characterized as conventional reversible photooxidants in their triplet states. The utility of these sensitizers is demonstrated by irreversibly generating the guanosine radical cation in buffered aqueous solution. PMID:24354634

  19. Transiently Produced Hypochlorite Is Responsible for the Irreversible Inhibition of Chlorite Dismutase

    PubMed Central

    2014-01-01

    Chlorite dismutases (Clds) are heme b-containing prokaryotic oxidoreductases that catalyze the reduction of chlorite to chloride with the concomitant release of molecular oxygen. Over time, they are irreversibly inactivated. To elucidate the mechanism of inactivation and investigate the role of the postulated intermediate hypochlorite, the pentameric chlorite dismutase of “Candidatus Nitrospira defluvii” (NdCld) and two variants (having the conserved distal arginine 173 exchanged with alanine and lysine) were recombinantly produced in Escherichia coli. Exchange of the distal arginine boosts the extent of irreversible inactivation. In the presence of the hypochlorite traps methionine, monochlorodimedone, and 2-[6-(4-aminophenoxy)-3-oxo-3H-xanthen-9-yl]benzoic acid, the extent of chlorite degradation and release of molecular oxygen is significantly increased, whereas heme bleaching and oxidative modifications of the protein are suppressed. Among other modifications, hypochlorite-mediated formation of chlorinated tyrosines is demonstrated by mass spectrometry. The data obtained were analyzed with respect to the proposed reaction mechanism for chlorite degradation and its dependence on pH. We discuss the role of distal Arg173 by keeping hypochlorite in the reaction sphere for O–O bond formation. PMID:24754261

  20. Emergent irreversibility and entanglement spectrum statistics.

    PubMed

    Chamon, Claudio; Hamma, Alioscia; Mucciolo, Eduardo R

    2014-06-20

    We study the problem of irreversibility when the dynamical evolution of a many-body system is described by a stochastic quantum circuit. Such evolution is more general than a Hamiltonian one, and since energy levels are not well defined, the well-established connection between the statistical fluctuations of the energy spectrum and irreversibility cannot be made. We show that the entanglement spectrum provides a more general connection. Irreversibility is marked by a failure of a disentangling algorithm and is preceded by the appearance of Wigner-Dyson statistical fluctuations in the entanglement spectrum. This analysis can be done at the wave-function level and offers an alternative route to study quantum chaos and quantum integrability.

  1. Entropy production in irreversible processes with friction.

    PubMed

    Bizarro, João P S

    2008-08-01

    Established expressions for entropy production in irreversible processes are generalized to include friction explicitly, as a source of irreversibility in the interaction between a system and its surroundings. The net amount of heat delivered to the system does not come now only from the reservoir, but may have an additional component coming from the work done against friction forces and dissipated as heat. To avoid ambiguities in interpreting the different contributions to entropy increase, the latter is also written in terms of the heat directly exchanged between the system and surroundings and of the fraction of frictional work that is lost in the system. PMID:18850816

  2. Efficiency of Rectification: Reversible vs. Irreversible Regimes

    NASA Astrophysics Data System (ADS)

    Sokolov, I. M.

    2002-11-01

    Both man-made locomotive devices and molecular motors use gears to transform a reciprocating motion into a directed one. One of the most common gears is a rectifier, a mechanically irreversible appliance. The maximal energetic efficiency of an isothermic gear is bounded by unity, as a consequence of the Second Law. However, approaching this ideal efficiency does not imply approaching reversibility. We discuss what properties of a rectifier mostly influence the transduction efficiency and show that an appliance which locks under backward force is just the one which can approach the ideal efficiency either in the reversible or in the irreversible regime.

  3. Time and Irreversibility in AN Accelerating Universe

    NASA Astrophysics Data System (ADS)

    Romero, Gustavo E.; Pérez, Daniela

    It is a remarkable fact that all processes occurring in the observable universe are irreversible, whereas the equations through which the fundamental laws of physics are formulated are invariant under time reversal. The emergence of irreversibility from the fundamental laws has been a topic of consideration by physicists, astronomers and philosophers since Boltzmann's formulation of his famous "H" theorem. In this paper, we shall discuss some aspects of this problem and its connection with the dynamics of spacetime, within the framework of modern cosmology. We conclude that the existence of cosmological horizons allows a coupling of the global state of the universe with the local events determined through electromagnetic processes.

  4. Pilot Decision-Making in Irreversible Emergencies

    ERIC Educational Resources Information Center

    Winter, Scott R.

    2013-01-01

    The purpose of this study was to determine if a reflexive learning treatment utilizing select case studies could enhance the decision-making of pilots who encounter an irreversible emergency. Participants, who consisted of members of the subject university's professional pilot program, were divided into either a control or experimental group and…

  5. Absorption media for irreversibly gettering thionyl chloride

    DOEpatents

    Buffleben, George; Goods, Steven H.; Shepodd, Timothy; Wheeler, David R.; Whinnery, Jr., LeRoy

    2002-01-01

    Thionyl chloride is a hazardous and reactive chemical used as the liquid cathode in commercial primary batteries. Contrary to previous thinking, ASZM-TEDA.RTM. carbon (Calgon Corporation) reversibly absorbs thionyl chloride. Thus, several candidate materials were examined as irreversible getters for thionyl chloride. The capacity, rate and effect of temperature were also explored. A wide variety of likely materials were investigated through screening experiments focusing on the degree of heat generated by the reaction as well as the material absorption capacity and irreversibility, in order to help narrow the group of possible getter choices. More thorough, quantitative measurements were performed on promising materials. The best performing getter was a mixture of ZnO and ASZM-TEDA.RTM. carbon. In this example, the ZnO reacts with thionyl chloride to form ZnCl.sub.2 and SO.sub.2. The SO.sub.2 is then irreversibly gettered by ASZM-TEDA.RTM. carbon. This combination of ZnO and carbon has a high capacity, is irreversible and functions effectively above -20.degree. C.

  6. Markov Chain Monte Carlo and Irreversibility

    NASA Astrophysics Data System (ADS)

    Ottobre, Michela

    2016-06-01

    Markov Chain Monte Carlo (MCMC) methods are statistical methods designed to sample from a given measure π by constructing a Markov chain that has π as invariant measure and that converges to π. Most MCMC algorithms make use of chains that satisfy the detailed balance condition with respect to π; such chains are therefore reversible. On the other hand, recent work [18, 21, 28, 29] has stressed several advantages of using irreversible processes for sampling. Roughly speaking, irreversible diffusions converge to equilibrium faster (and lead to smaller asymptotic variance as well). In this paper we discuss some of the recent progress in the study of nonreversible MCMC methods. In particular: i) we explain some of the difficulties that arise in the analysis of nonreversible processes and we discuss some analytical methods to approach the study of continuous-time irreversible diffusions; ii) most of the rigorous results on irreversible diffusions are available for continuous-time processes; however, for computational purposes one needs to discretize such dynamics. It is well known that the resulting discretized chain will not, in general, retain all the good properties of the process that it is obtained from. In particular, if we want to preserve the invariance of the target measure, the chain might no longer be reversible. Therefore iii) we conclude by presenting an MCMC algorithm, the SOL-HMC algorithm [23], which results from a nonreversible discretization of a nonreversible dynamics.

  7. A new microscopic level of irreversibility

    SciTech Connect

    Prigogine, I.

    1987-01-01

    In this paper, the non-exponential decay is analyzed with the help of simple computer experiments performed by T. Petrosky, simulating classical radiation damping. The non-exponential decay is studied and shown to depend on the preparation of the system. However, whatever the initial preparation, the system reaches the decay predicted by classical radiation theory after a short time we call the Zeno's time. The similitude of Petrosky's results with computer experiments for the approach to equilibrium in many-body systems is emphasized. However, while there one deals with times which are multiple of the relaxation time, the irreversibility manifest in radiation theory occurs always over a much shorter time scale, the Zeno's time. In atomic systems, this would be a time order of 10/sup /minus/18/ seconds. These results are of great interest for the understanding of the microscopic mechanism of radiation. Let us consider a charged oscillator. In a first stage, this oscillator has to produce the field oscillators to which it may transfer energy through the usual resonance mechanism. Radiation appears therefore as a kind of non linear autocatalytic process, involving a self-organization mechanism. The behavior during the Zeno period can be explained easily in terms of subdynamics as introduced by the Brussel's group. We see that there is no transition from reversibility to irreversibility. Irreversible processes start at the very moment at which the system is prepared. It is important to stress that an unstable particle is itself the result of irreversible processes. As a result, an unstable particle (or an excited atomic state) can no more be described in terms of wave functions, as irreversible processes are not included in Schroedinger's equation. 14 refs., 3 figs.

  8. Thermal inactivation of enzymes and pathogens in biosamples for MS analysis.

    PubMed

    Ahnoff, Martin; Cazares, Lisa H; Sköld, Karl

    2015-01-01

    Protein denaturation is the common basis for enzyme inactivation and inactivation of pathogens, necessary for preservation and safe handling of biosamples for downstream analysis. While heat-stabilization technology has been used in proteomic and peptidomic research since its introduction in 2009, the advantages of using the technique for simultaneous pathogen inactivation have only recently been addressed. The time required for enzyme inactivation by heat (≈1 min) is short compared with chemical treatments, and inactivation is irreversible in contrast to freezing. Heat stabilization thus facilitates mass spectrometric studies of biomolecules with a fast conversion rate, and expands the chemical space of potential biomarkers to include more short-lived entities, such as phosphorylated proteins, in tissue samples as well as whole-blood (dried blood sample) samples. PMID:26295989

  9. Role of NADH oxidase in the oxidative inactivation of Streptococcus salivarius fructosyltransferase.

    PubMed Central

    Abbe, K; Takahashi-Abbe, S; Schoen, R A; Wittenberger, C L

    1986-01-01

    A cell-associated fructosyltransferase produced by Streptococcus salivarius was irreversibly inactivated in a time-dependent manner when resting or permeabilized cell suspensions were incubated with low concentrations (less than 1.0 microM) of copper. In addition to copper, the inactivation was dependent on oxygen and on a fermentable carbon source (endogenous intracellular polysaccharide or glucose). In starved, permeabilized cell suspensions, the fermentable carbon source could be replaced by NADH but not by NADPH or ATP. Of several other S. salivarius enzymes tested, only fructosyltransferase was inactivated under these conditions. The available evidence indicated that NADH oxidase is the enzyme responsible for fructosyltransferase inactivation. Results from oxygen radical scavenger studies implicated one or more species of oxygen radicals and hydrogen peroxide in the inactivation reaction. PMID:3759237

  10. Inactivated pepsin inhibits neutrophil activation by Fcgamma-receptor-dependent and independent stimuli.

    PubMed

    Kustiawan, Iwan; Derksen, Ninotska; Rispens, Theo

    2016-08-01

    Pepsin is widely used to produce F(ab')2 fragments of immunoglobulin G (IgG). In many cases, at least part of the pepsin will remain present in the F(ab')2 preparation, albeit in (irreversibly) inactivated form. Here we report on a potent immunomodulatory effect of irreversibly inactivated pepsin on activated human neutrophils. Degranulation, induced by coated IgG or via cytochalasin B/N-formyl-Met-Leu-Phe, was measured by quantifying elastase release, and was found to be inhibited in a dose-dependent manner by inactivated pepsin. Since a number of intravenous immunoglobulin (IVIg) products are also treated by limited digestion with pepsin, we investigated if pepsin would be present in quantities large enough to inhibit neutrophil activation. The amounts of pepsin detected in three different pepsin-treated IVIg products were found to be too low to induce an effect, at least in an in vitro setting. PMID:27368805

  11. Mechanism-Based Inactivation of Ammonia Monooxygenase in Nitrosomonas europaea by Allylsulfide

    PubMed Central

    Juliette, Lisa Y.; Hyman, Michael R.; Arp, Daniel J.

    1993-01-01

    Allylsulfide caused an irreversible inactivation of ammonia monooxygenase (AMO) activity (ammonia-dependent O2 uptake) in Nitrosomonas europaea. The hydroxylamine oxidoreductase activity (hydrazine-dependent O2 uptake) of cells was unaffected by allylsulfide. Anaerobic conditions or the presence of allylthiourea, a reversible noncompetitive AMO inhibitor, protected AMO from inactivation by allylsulfide. Ammonia did not protect AMO from inactivation by allylsulfide but instead increased the rate of inactivation. The inactivation of AMO followed pseudo-first-order kinetics, but the observed rates did not saturate with increasing allylsulfide concentrations. The time course of recovery of AMO-dependent nitrite production after complete inactivation by allylsulfide required de novo protein synthesis. Incubation of cells with allylsulfide prevented the 14C label from 14C2H2 (a suicide mechanism-based inactivator of AMO) from being incorporated into the 27-kDa polypeptide of AMO. Some compounds structurally related to allylsulfide were unable to inactivate AMO. We conclude that allylsulfide is a specific, mechanism-based inactivator of AMO in N. europaea. PMID:16349087

  12. Multiscale multifractal time irreversibility analysis of stock markets

    NASA Astrophysics Data System (ADS)

    Jiang, Chenguang; Shang, Pengjian; Shi, Wenbin

    2016-11-01

    Time irreversibility is one of the most important properties of nonstationary time series. Complex time series often demonstrate even multiscale time irreversibility, such that not only the original but also coarse-grained time series are asymmetric over a wide range of scales. We study the multiscale time irreversibility of time series. In this paper, we develop a method called multiscale multifractal time irreversibility analysis (MMRA), which allows us to extend the description of time irreversibility to include the dependence on the segment size and statistical moments. We test the effectiveness of MMRA in detecting multifractality and time irreversibility of time series generated from delayed Henon map and binomial multifractal model. Then we employ our method to the time irreversibility analysis of stock markets in different regions. We find that the emerging market has higher multifractality degree and time irreversibility compared with developed markets. In this sense, the MMRA method may provide new angles in assessing the evolution stage of stock markets.

  13. Irreversible enzyme inhibition kinetics and drug-drug interactions.

    PubMed

    Mohutsky, Michael; Hall, Stephen D

    2014-01-01

    This chapter describes the types of irreversible inhibition of drug-metabolizing enzymes and the methods commonly employed to quantify the irreversible inhibition and subsequently predict the extent and time course of clinically important drug-drug interactions.

  14. Structure of suicide-inactivated. beta. -hydroxydecanoyl-thioester dehydrase

    SciTech Connect

    Schwab, J.M.; Ho, C.K.; Li, W.B.; Townsend, C.A.; Salituro, G.M.

    1986-05-01

    ..beta..-Hydroxydecanoylthioester dehydrase, the key enzyme in biosynthesis of unsaturated fatty acids under anaerobic conditions, equilibrates thioesters of (R)-3-hydroxydecanoic acid, E-2-decenoic acid, and Z-3-decenoic acid. Dehydrase is irreversibly inactivated by the N-acetylcysteamine thioester of 3-decynoic acid (3-decynoyl-NAC), via dehydrase-catalyzed isomerization to 2,3-decadienoyl-NAC. To probe the relationship between normal catalysis and suicide inactivation, the structure of the inactivated enzyme has been studied. 3-(2-/sup 13/C)Decynoyl-NAC was synthesized and incubated with dehydrase. /sup 13/C NMR showed that attack of 2,3-decadienoyl-NAC by the active site histidine gives 3-histidinyl-3-decenoyl-NAC, which slowly rearranges to the more stable ..delta../sup 2/ isomer. Model histidine-allene adducts have been made and characterized. Analysis of NMR data show that the C=C configuration of the decenoyl moiety of enzyme-bound inactivator is E. The suggestion that the mechanism of dehydrase inactivation parallels its normal mechanism of action is supported these findings.

  15. Inactivation of Anopheles gambiae Glutathione Transferase ε2 by Epiphyllocoumarin

    PubMed Central

    Marimo, Patience; Hayeshi, Rose; Mukanganyama, Stanley

    2016-01-01

    Glutathione transferases (GSTs) are part of a major family of detoxifying enzymes that can catalyze the reductive dehydrochlorination of dichlorodiphenyltrichloroethane (DDT). The delta and epsilon classes of insect GSTs have been implicated in conferring resistance to this insecticide. In this study, the inactivation of Anopheles gambiae GSTε2 by epiphyllocoumarin (Tral 1) was investigated. Recombinant AgGSTε2 was expressed in Escherichia coli cells containing a pET3a-AGSTε2 plasmid and purified by affinity chromatography. Tral 1 was shown to inactivate GSTε2 both in a time-dependent manner and in a concentration-dependent manner. The half-life of GSTε2 in the presence of 25 μM ethacrynic acid (ETA) was 22 minutes and with Tral 1 was 30 minutes, indicating that Tral 1 was not as efficient as ETA as an inactivator. The inactivation parameters kinact and KI were found to be 0.020 ± 0.001 min−1 and 7.5 ± 2.1 μM, respectively, after 90 minutes of incubation. Inactivation of GSTε2 by Tral 1 implies that Tral 1 covalently binds to this enzyme in vitro and would be expected to exhibit time-dependent effects on the enzyme in vivo. Tral 1, therefore, would produce irreversible effects when used together with dichlorodiphenyltrichloroethane (DDT) in malaria control programmes where resistance is mediated by GSTs. PMID:26925266

  16. Inactivation of calcium current in bull-frog atrial myocytes.

    PubMed Central

    Campbell, D L; Giles, W R; Hume, J R; Shibata, E F

    1988-01-01

    1. A single-microelectrode technique has been used to study the voltage dependence and the kinetics of inactivation and reactivation of a tetrodotoxin-resistant inward current (ICa) in single cells from bull-frog atrium. 2. In most cases the kinetics of both inactivation and reactivation can be well described as a single-exponential process. 3. Several different observations indicate that inactivation of ICa in these cells is controlled by both voltage-dependent and current-dependent processes, as has been demonstrated previously in heart (Kass & Sanguinetti, 1984; Lee, Marban & Tsien, 1985) and in other tissues (Hagiwara & Byerly, 1981; Tsien, 1983; Eckert & Chad, 1984). 4. Evidence in favour of a voltage-dependent inactivation mechanism included: (a) In paired-pulse measurements of steady-state inactivation ('f infinity') a 'conventional' steady-state f infinity vs. membrane potential (Vm) relationship was obtained in the range of membrane potentials from -60 to 0 mV. (b) Increasing [Ca2+]o from 2.5 to 7.5 mM, which resulted in a 2-3-fold increase in ICa, did not produce any significant increase in the amount of inactivation. (c) Using a 'gapped' double-pulse protocol non-monotonic U-shaped inactivation relationships were obtained, i.e. positive to approximately +20 mV some removal of inactivation occurred. However, f never approached a value near 1.00 at very depolarized potentials; it reached a maximum between 0.5 and 0.6. (d) In constant [Ca2+]o and at fixed Vm, the kinetics of ICa inactivation were independent of peak size of ICa. This was demonstrated by: (i) varying the holding potential (-90 to -30 mV), (ii) using paired-pulse 'recovery' protocols, and (iii) partial block by La3+ (1-10 microM) and Cd2+ (0.1 mM). (e) Influx of Ca2+ ions was not an obligatory prerequisite for development of inactivation. In all ionic conditions (Ca2+, Sr2+, Ba2+, Na+-free and Ca2+-free Ringer solutions) currents displayed inactivation phenomena, although the extent and

  17. Catalase inactivation following photosensitization with tetrasulfonated metallophthalocyanines.

    PubMed

    Gantchev, T G; van Lier, J E

    1995-07-01

    Catalase (CAT) in solution or incorporated in erythrocytes and K562 leukemic cells is inactivated during photosensitization with tetrasulfonated metallophthalocyanines (MePcS4). The effect of added scavengers and D2O showed that both singlet oxygen and free radical species are involved in this process. Evidence was found that direct interactions of ground or excited-stated photosensitizer with CAT are not responsible for CAT inactivation. Specific techniques to probe early damage to the CAT structure involved optical and EPR spectroscopy, HPLC and polyacrylamide gel electrophoresis analyses. Different primary events of photosensitized protein damage included oxidation of cysteine residues as well as other amino acids, as demonstrated by the formation of carbon-centered free radicals and the loss of absorbance at lambda = 275 nm. In parallel, we detected degradation of the CAT heme groups, accompanied by release of Fe(II) ions in solution. These combined phenomena initiate cross-linkages between CAT subunits and subsequent degradation of the protein with formation of irreversible aggregates in solution. Phthalocyanine-mediated photoinactivation of cell-bound CAT results in loss of protection against accumulating H2O2, providing an additional pathway of phototoxicity. PMID:7638256

  18. Inactivation of prion infectivity by ionizing rays

    NASA Astrophysics Data System (ADS)

    Gominet, M.; Vadrot, C.; Austruy, G.; Darbord, J. C.

    2007-11-01

    Inactivation of prion deposits on medical devices or prion contamination in pharmaceutical raw materials is considered as impossible by using gamma irradiation. Early, the guideline WHO/CDS/CSR/APH/2000 has described irradiation as an ineffective process. But, in 2003, S. Miekka et al. noted radiation inactivation of prions in a particular application to purify human albumin, shown by the physical denaturation of the infectious protein (PrP). The aim of our study was to determine the inactivation of prions with a scrapie model (strain C506M3) by irradiating standardised preparations. Results: Gamma irradiation was partially effective, showing a 4-5 log reduction on exposure to 50 kGy. A characteristic effect-dose curve was not observed (25, 50 and 100 kGy), only an increase in the incubation period of the murine disease (229 days with 25 kGy to 290 days with 100 kGy) compared with 170 days without irradiation. Since the inactivation was not a total one, the observed effect is significant. It is proposed that further work be undertaken with the model to investigate the application of gamma radiation known levels of prion contamination.

  19. Impact of HIV-1 Membrane Cholesterol on Cell-Independent Lytic Inactivation and Cellular Infectivity.

    PubMed

    Kalyana Sundaram, Ramalingam Venkat; Li, Huiyuan; Bailey, Lauren; Rashad, Adel A; Aneja, Rachna; Weiss, Karl; Huynh, James; Bastian, Arangaserry Rosemary; Papazoglou, Elisabeth; Abrams, Cameron; Wrenn, Steven; Chaiken, Irwin

    2016-01-26

    Peptide triazole thiols (PTTs) have been found previously to bind to HIV-1 Env spike gp120 and cause irreversible virus inactivation by shedding gp120 and lytically releasing luminal capsid protein p24. Since the virions remain visually intact, lysis appears to occur via limited membrane destabilization. To better understand the PTT-triggered membrane transformation involved, we investigated the role of envelope cholesterol on p24 release by measuring the effect of cholesterol depletion using methyl beta-cyclodextrin (MβCD). An unexpected bell-shaped response of PTT-induced lysis to [MβCD] was observed, involving lysis enhancement at low [MβCD] vs loss of function at high [MβCD]. The impact of cholesterol depletion on PTT-induced lysis was reversed by adding exogenous cholesterol and other sterols that support membrane rafts, while sterols that do not support rafts induced only limited reversal. Cholesterol depletion appears to cause a reduced energy barrier to lysis as judged by decreased temperature dependence with MβCD. Enhancement/replenishment responses to [MβCD] also were observed for HIV-1 infectivity, consistent with a similar energy barrier effect in the membrane transformation of virus cell fusion. Overall, the results argue that cholesterol in the HIV-1 envelope is important for balancing virus stability and membrane transformation, and that partial depletion, while increasing infectivity, also makes the virus more fragile. The results also reinforce the argument that the lytic inactivation and infectivity processes are mechanistically related and that membrane transformations occurring during lysis can provide an experimental window to investigate membrane and protein factors important for HIV-1 cell entry.

  20. Mechanisms of inactivation of lipoxygenases by phenidone and BW755C.

    PubMed

    Cucurou, C; Battioni, J P; Thang, D C; Nam, N H; Mansuy, D

    1991-09-17

    Inhibition of soybean lipoxygenase (L-1) and potato 5-lipoxygenase (5-PLO) by the pyrazoline derivatives phenidone and BW755C only occurs after oxidation of these compounds by the peroxidase-like activity of the lipoxygenases. There is a clear relationship between this oxidation and the irreversible inactivation of L-1. The final product of phenidone oxidation by L-1, 4,5-didehydrophenidone, is not responsible of this inactivation, but the species derived from a one-electron oxidation of phenidone plays a key role in L-1 inactivation. In the absence of O2, inactivation of 1 mol of L-1 occurs after the oxidation of 34 mol of phenidone and the covalent binding of 0.8 mol of phenidone-derived metabolite(s) to L-1. In the presence of O2, inactivation of 1 mol of L-1 occurs already after oxidation of 11 mol of phenidone and only involves the covalent binding of 0.4 mol of phenidone-derived metabolite(s) to L-1. A mechanism is proposed for L-1 inactivation by phenidone, which involves the irreversible binding of a phenidone metabolite to the protein and the oxidation of an L-1 amino acid residue (in the presence of O2). PMID:1654081

  1. Unfolding and inactivation of proteins by counterions in protein-nanoparticles interaction.

    PubMed

    Ghosh, Goutam; Gaikwad, Pallavi S; Panicker, Lata; Nath, Bimalendu B; Mukhopadhyaya, Rita

    2016-09-01

    In this work, the structure and activity of proteins; such as, hen egg lysozyme (HEWL) and calf intestine alkaline phosphatase (CIAP); have been investigated after incubation with surface coated iron oxide nanoparticles (IONPs) in water. IONPs were coated with counterions bound charge-ligands and were named as the charge-ligand counterions iron oxide nanoparticles (CLC-IONPs). The coating was done with tri-lithium citrate (TLC) and tri-potassium citrate (TKC) to have negative surface charge of CLC-IONPs and Li(+) and K(+), respectively, as counterions. To have positive surface charge, IONPs were coated with cetylpyridinium chloride (CPC) and cetylpyridinium iodide (CPI) having Cl(-) and I(-), respectively, as counterions. The secondary structure of proteins was measured using far ultraviolet circular dichroism (CD) spectroscopy which showed that both proteins were irreversibly unfolded after incubation with CLC-IONPs. The unfolded proteins were seen to be functionally inactive, as confirmed through their activity assays, i.e., HEWL with Escherichia coli (E. coli) and CIAP with para-nitrophenyl phosphate (pNPP). Additionally, we have observed that monomeric hemoglobin (Hb) from radio-resistant insect Chironomus ramosus (ChHb) was also partially unfolded upon interaction with CLC-IONPs. This work clearly shows the role of counterions in protein inactivation via protein-nanoparticles interaction and, therefore, CLC-IONPs could be used for therapeutic purpose. PMID:27182654

  2. Structural changes associated with poliovirus inactivation in soil.

    PubMed Central

    Yeager, J G; O'Brien, R T

    1979-01-01

    The loss of infectivity of poliovirus in moist and dried soils was a result of irreversible damage to the virus particles. The damage included (i) dissociation of viral genomes and capsids and (ii) degradation of viral ribonucleic acid (RNA) in the soil environment. Under drying conditions, capsid components could not be recovered from the soils. Further studies in sterile soils indicated that, under moist conditions, the viral RNA was probably damaged before dissociation from the capsid. However, in sterile, dried soil, RNA genomes were recovered largely intact from the soil. These results suggest that polioviruses are inactivated by different mechanisms in moist and drying soils. PMID:231938

  3. Calcium-binding parameter of Bacillus amyloliquefaciens alpha-amylase determined by inactivation kinetics.

    PubMed Central

    Tanaka, Atsushi; Hoshino, Eiichi

    2002-01-01

    The irreversible thermal inactivation and the thermodynamics of calcium ion binding of Bacillus amyloliquefaciens alpha-amylase in the absence of substrates were studied. The enzyme inactivation on heating was apparently followed by first-order kinetics. The enzyme was stabilized with an increased concentration of calcium ion and thus the inactivation was highly dependent on the state of calcium binding. The activation parameter for the inactivation suggests an unfolding of the enzyme protein upon heating. Values of both the activation enthalpy and entropy were increased with a higher calcium ion concentration. An inactivation kinetic model is based on the assumption of a two-stage unfolding transition in which the bivalent ion dissociation occurs in the first step followed by the secondary structural unfolding. This simple kinetic model provides both a qualitative and quantitative interpretation of calcium ion binding to the enzyme and its effect on the inactivation properties. The specific approximations of the kinetic model were strictly followed in the analysis to calculate the apparent inactivation rate at each calcium ion concentration in terms of the calcium-binding parameters. The enthalpy and entropy changes for the calcium ion binding were calculated to be -149 kJ/mol and -360 J.mol(-1).K(-1) respectively and these values suggest a strong enthalpic affinity for the bivalent ion binding to the enzyme protein. The thermodynamical interpretation attempts to provide clear relations between the terms of an apparent inactivation rate and the calcium binding. PMID:12049626

  4. Intracranial nonthermal irreversible electroporation: in vivo analysis.

    PubMed

    Garcia, Paulo A; Rossmeisl, John H; Neal, Robert E; Ellis, Thomas L; Olson, John D; Henao-Guerrero, Natalia; Robertson, John; Davalos, Rafael V

    2010-07-01

    Nonthermal irreversible electroporation (NTIRE) is a new minimally invasive technique to treat cancer. It is unique because of its nonthermal mechanism of tumor ablation. Intracranial NTIRE procedures involve placing electrodes into the targeted area of the brain and delivering a series of short but intense electric pulses. The electric pulses induce irreversible structural changes in cell membranes, leading to cell death. We correlated NTIRE lesion volumes in normal brain tissue with electric field distributions from comprehensive numerical models. The electrical conductivity of brain tissue was extrapolated from the measured in vivo data and the numerical models. Using this, we present results on the electric field threshold necessary to induce NTIRE lesions (495-510 V/cm) in canine brain tissue using 90 50-mus pulses at 4 Hz. Furthermore, this preliminary study provides some of the necessary numerical tools for using NTIRE as a brain cancer treatment. We also computed the electrical conductivity of brain tissue from the in vivo data (0.12-0.30 S/m) and provide guidelines for treatment planning and execution. Knowledge of the dynamic electrical conductivity of the tissue and electric field that correlates to lesion volume is crucial to ensure predictable complete NTIRE treatment while minimizing damage to surrounding healthy tissue. PMID:20668843

  5. On phylogenetic tests of irreversible evolution.

    PubMed

    Goldberg, Emma E; Igić, Boris

    2008-11-01

    "Dollo's law" states that, following loss, a complex trait cannot reevolve in an identical manner. Although the law has previously fallen into disrepute, it has only recently been challenged with statistical phylogenetic methods. We employ simulation studies of an irreversible binary character to show that rejections of Dollo's law based on likelihood-ratio tests of transition rate constraints or on reconstructions of ancestral states are frequently incorrect. We identify two major causes of errors: incorrect assignment of root state frequencies, and neglect of the effect of the character state on rates of speciation and extinction. Our findings do not necessarily overturn the conclusions of phylogenetic studies claiming reversals, but we demonstrate devastating flaws in the methods that are the foundation of all such studies. Furthermore, we show that false rejections of Dollo's law can be reduced by the use of appropriate existing models and model selection procedures. More powerful tests of irreversibility require data beyond phylogenies and character states of extant taxa, and we highlight empirical work that incorporates additional information.

  6. Probabilistic Gompertz model of irreversible growth.

    PubMed

    Bardos, D C

    2005-05-01

    Characterizing organism growth within populations requires the application of well-studied individual size-at-age models, such as the deterministic Gompertz model, to populations of individuals whose characteristics, corresponding to model parameters, may be highly variable. A natural approach is to assign probability distributions to one or more model parameters. In some contexts, size-at-age data may be absent due to difficulties in ageing individuals, but size-increment data may instead be available (e.g., from tag-recapture experiments). A preliminary transformation to a size-increment model is then required. Gompertz models developed along the above lines have recently been applied to strongly heterogeneous abalone tag-recapture data. Although useful in modelling the early growth stages, these models yield size-increment distributions that allow negative growth, which is inappropriate in the case of mollusc shells and other accumulated biological structures (e.g., vertebrae) where growth is irreversible. Here we develop probabilistic Gompertz models where this difficulty is resolved by conditioning parameter distributions on size, allowing application to irreversible growth data. In the case of abalone growth, introduction of a growth-limiting biological length scale is then shown to yield realistic length-increment distributions.

  7. Irreversible heavy chain transfer to chondroitin.

    PubMed

    Lauer, Mark E; Hascall, Vincent C; Green, Dixy E; DeAngelis, Paul L; Calabro, Anthony

    2014-10-17

    We have recently demonstrated that the transfer of heavy chains (HCs) from inter-α-inhibitor, via the enzyme TSG-6 (tumor necrosis factor-stimulated gene 6), to hyaluronan (HA) oligosaccharides is an irreversible event in which subsequent swapping of HCs between HA molecules does not occur. We now describe our results of HC transfer experiments to chondroitin sulfate A, chemically desulfated chondroitin, chemoenzymatically synthesized chondroitin, unsulfated heparosan, heparan sulfate, and alginate. Of these potential HC acceptors, only chemically desulfated chondroitin and chemoenzymatically synthesized chondroitin were HC acceptors. The kinetics of HC transfer to chondroitin was similar to HA. At earlier time points, HCs were more widely distributed among the different sizes of chondroitin chains. As time progressed, the HCs migrated to lower molecular weight chains of chondroitin. Our interpretation is that TSG-6 swaps the HCs from the larger, reversible sites on chondroitin chains, which function as HC acceptors, onto smaller chondroitin chains, which function as irreversible HC acceptors. HCs transferred to smaller chondroitin chains were unable to be swapped off the smaller chondroitin chains and transferred to HA. HCs transferred to high molecular weight HA were unable to be swapped onto chondroitin. We also present data that although chondroitin was a HC acceptor, HA was the preferred acceptor when chondroitin and HA were in the same reaction mixture.

  8. Magnetic Irreversibility in VO2/Ni Bilayers

    NASA Astrophysics Data System (ADS)

    de La Venta, Jose; Lauzier, Josh; Sutton, Logan

    The temperature dependence of the coercivity and magnetization of VO2/Ni bilayers was studied. VO2 exhibits a well-known Structural Phase Transition (SPT) at 330-340 K, from a low temperature monoclinic (M) to a high temperature rutile (R) structure. The SPT of VO2 induces an inverse magnetoelastic effect that strongly modifies the coercivity and magnetization of the Ni films. In addition, the growth conditions allow tuning of the magnetic properties. Ni films deposited on top of VO2 (M) show an irreversible change in the coercivity after the first cycle through the high temperature phase, with a corresponding change in the surface morphology of VO2. On the other hand, the Ni films grown on top of VO2 (R) do not show this irreversibility. These results indicate that properties of magnetic films are strongly affected by the strain induced by materials that undergo SPT and that it is possible to control the magnetic properties by tuning the growth conditions.

  9. Irreversible sediment formation in green tea infusions.

    PubMed

    Xu, Yong-Quan; Chen, Gen-Sheng; Wang, Qiu-Shuang; Yuan, Hai-Bo; Feng, Chun-Hong; Yin, Jun-Feng

    2012-03-01

    The formation of irreversible tea sediment (IRS) and its chemical components in green tea infusions were investigated. The results showed that the amounts of IRS in the green tea infusions from various tea cultivars ranged from 0.10 to 1.47 mg/mL. The amount of IRS was influenced remarkably by the chemical components in the green tea infusion. Principal component analysis and regression analysis indicated that gallated catechins, Mn, Ca, caffeine, Na, and (-)-gallocatechin gallate (GCG) were the principal components. IRS (mg/mL) = -4.226 + 0.275 gallated catechins + 79.551 Na + 7.321 Mn + 21.055 Ca + 0.513 caffeine - 0.129 GCG (R2 = 0.697). The contents of the main chemical components in the reversible tea sediment (RTS) and IRS were markedly different, especially the minerals. Large amount of minerals participated in the formation of irreversible green tea sediment. The amount of IRS increased with the extraction temperature. PMID:22329921

  10. Nucleation of a new phase on a surface that is changing irreversibly with time.

    PubMed

    Sear, Richard P

    2014-02-01

    Nucleation of a new phase almost always starts at a surface. This surface is almost always assumed not to change with time. However, surfaces can roughen, partially dissolve, and change chemically with time. Each of these irreversible changes will change the nucleation rate at the surface, resulting in a time-dependent nucleation rate. Here we use a simple model to show that partial surface dissolution can qualitatively change the nucleation process in a way that is testable in experiment. The changing surface means that the nucleation rate is increasing with time. There is an initial period during which no nucleation occurs, followed by relatively rapid nucleation. PMID:25353480

  11. Photodynamic inactivation of the Na,K-ATPase occurs via different pathways.

    PubMed

    Killig, F; Stark, G; Apell, H J

    2004-08-01

    The photodynamic, i.e., the light-induced, inactivation of the Na,K-ATPase in the presence of the sensitizer rose bengal was studied under different conditions. The shape of inactivation curves of the enzyme activity was analyzed as well as partial reactions of the pump cycle. Both experimental approaches showed the existence of two different time constants of inactivation of the ion pump, which reflect two pathways of a photodynamic modification. This is supported by the following observations: (1) The amplitude of the initial fast decay of enzyme activity was enhanced in the presence of D2O and reduced in the presence of the singlet oxygen scavenger imidazole. (Similar results were found for the SR Ca-ATPase.) (2) Contrary to the fast enzyme inactivation the slow process shows an inverse dose-rate behavior. (3) Inactivation of the partial reactions of Na+ -binding and of K+-binding to the membrane domain of the Na,K-ATPase showed only a single time constant, which corresponded to the slower time constant of enzyme inactivation. In the presence of high concentrations of singlet oxygen the fast time constant dominated the inactivation of the ATP-induced partial reaction for which the cytoplasmic domains of the enzyme play an important role. The data support the conclusion that fast inactivation is due to modification of the cytoplasmic domains and slow inactivation due to modifications of the membrane domain of the ion pumps.

  12. Radiation inactivation study of aminopeptidase: probing the active site

    NASA Astrophysics Data System (ADS)

    Jamadar, V. K.; Jamdar, S. N.; Mohan, Hari; Dandekar, S. P.; Harikumar, P.

    2004-04-01

    Ionizing radiation inactivated purified chicken intestinal aminopeptidase in media saturated with gases in the order N 2O>N 2>air. The D 37 values in the above conditions were 281, 210 and 198 Gy, respectively. OH radical scavengers such as t-butanol and isopropanol effectively nullified the radiation-induced damage in N 2O. The radicals (SCN) 2•-, Br 2•- and I 2•- inactivated the enzyme, pointing to the involvement of aromatic amino acids and cysteine in its catalytic activity. The enzyme exhibited fluorescence emission at 340 nm which is characteristic of tryptophan. The radiation-induced loss of activity was accompanied by a decrease in the fluorescence of the enzyme suggesting a predominant influence on tryptophan residues. The enzyme inhibition was associated with a marked increase in the Km and a decrease in the Vmax and kcat values, suggesting an irreversible alteration in the catalytic site. The above observations were confirmed by pulse radiolysis studies.

  13. Inactivation of phospholipase A2 by naturally occurring biflavonoid, ochnaflavone.

    PubMed

    Chang, H W; Baek, S H; Chung, K W; Son, K H; Kim, H P; Kang, S S

    1994-11-30

    Ochnaflavone, a medicinal herb product isolated from Lonicera japonica, strongly inhibited rat platelet phospholipase A2 (IC50, about 3 microM). Inactivation was concentration and pH dependent (maximum inactivation occurred between pH 9.0 and 10.0). Ochnaflavone inhibited the enzyme by a noncompetitive manner, with the apparent Ki value of 3 x 10(-5) M. Reversibility was studied directly by dialysis method; the inhibition was irreversible. In addition, the inhibitory activity of ochnaflavone is rather specific against group II phospholipase A2 than group I phospholipase A2 (IC50, about 20 microM). Addition of excess Ca2+ concentration up to 8 mM did not antagonize the inhibitory activity of ochnaflavone. These results indicate that the inhibition of phospholipase A2 by ochnaflavone may result from direct interaction with the enzyme.

  14. Irreversible electroporation: state of the art

    PubMed Central

    Wagstaff, Peter GK; Buijs, Mara; van den Bos, Willemien; de Bruin, Daniel M; Zondervan, Patricia J; de la Rosette, Jean JMCH; Laguna Pes, M Pilar

    2016-01-01

    The field of focal ablative therapy for the treatment of cancer is characterized by abundance of thermal ablative techniques that provide a minimally invasive treatment option in selected tumors. However, the unselective destruction inflicted by thermal ablation modalities can result in damage to vital structures in the vicinity of the tumor. Furthermore, the efficacy of thermal ablation intensity can be impaired due to thermal sink caused by large blood vessels in the proximity of the tumor. Irreversible electroporation (IRE) is a novel ablation modality based on the principle of electroporation or electropermeabilization, in which electric pulses are used to create nanoscale defects in the cell membrane. In theory, IRE has the potential of overcoming the aforementioned limitations of thermal ablation techniques. This review provides a description of the principle of IRE, combined with an overview of in vivo research performed to date in the liver, pancreas, kidney, and prostate. PMID:27217767

  15. Exergetic sustainability evaluation of irreversible Carnot refrigerator

    NASA Astrophysics Data System (ADS)

    Açıkkalp, Emin

    2015-10-01

    Purpose of this paper is to assess irreversible refrigeration cycle by using exergetic sustainability index. In literature, there is no application of exergetic sustainability index for the refrigerators and, indeed, this index has not been derived for refrigerators. In this study, exergetic sustainability indicator is presented for the refrigeration cycle and its relationships with other thermodynamics parameters including COP, exergy efficiency, cooling load, exergy destruction, ecological function and work input are investigated. Calculations are conducted for endoreversible and reversible cycles and then results obtained from the ecological function are compared. It is found that exergy efficiency, exergetic sustainable index reduce 47.595% and 59.689% and rising at the COP is 99.888% is obtained for endoreversible cycle. Similarly, exergy efficiency and exergetic sustainability index reduce 90.163% and 93.711% and rising of the COP is equal to 99.362%.

  16. Irreversible electroporation: state of the art.

    PubMed

    Wagstaff, Peter Gk; Buijs, Mara; van den Bos, Willemien; de Bruin, Daniel M; Zondervan, Patricia J; de la Rosette, Jean Jmch; Laguna Pes, M Pilar

    2016-01-01

    The field of focal ablative therapy for the treatment of cancer is characterized by abundance of thermal ablative techniques that provide a minimally invasive treatment option in selected tumors. However, the unselective destruction inflicted by thermal ablation modalities can result in damage to vital structures in the vicinity of the tumor. Furthermore, the efficacy of thermal ablation intensity can be impaired due to thermal sink caused by large blood vessels in the proximity of the tumor. Irreversible electroporation (IRE) is a novel ablation modality based on the principle of electroporation or electropermeabilization, in which electric pulses are used to create nanoscale defects in the cell membrane. In theory, IRE has the potential of overcoming the aforementioned limitations of thermal ablation techniques. This review provides a description of the principle of IRE, combined with an overview of in vivo research performed to date in the liver, pancreas, kidney, and prostate.

  17. Performance of an irreversible quantum Carnot engine with spin 12.

    PubMed

    Wu, Feng; Chen, Lingen; Wu, Shuang; Sun, Fengrui; Wu, Chih

    2006-06-01

    The purpose of this paper is to investigate the effect of quantum properties of the working medium on the performance of an irreversible Carnot cycle with spin 12. The optimal relationship between the dimensionless power output P* versus the efficiency eta for the irreversible quantum Carnot engine with heat leakage and other irreversible losses is derived. Especially, the performances of the engine at low temperature limit and at high temperature limit are discussed.

  18. Heat-induced Irreversible Denaturation of the Camelid Single Domain VHH Antibody Is Governed by Chemical Modifications

    PubMed Central

    Akazawa-Ogawa, Yoko; Takashima, Mizuki; Lee, Young-Ho; Ikegami, Takahisa; Goto, Yuji; Uegaki, Koichi; Hagihara, Yoshihisa

    2014-01-01

    The variable domain of camelid heavy chain antibody (VHH) is highly heat-resistant and is therefore ideal for many applications. Although understanding the process of heat-induced irreversible denaturation is essential to improve the efficacy of VHH, its inactivation mechanism remains unclear. Here, we showed that chemical modifications predominantly governed the irreversible denaturation of VHH at high temperatures. After heat treatment, the activity of VHH was dependent only on the incubation time at 90 °C and was insensitive to the number of heating (90 °C)-cooling (20 °C) cycles, indicating a negligible role for folding/unfolding intermediates on permanent denaturation. The residual activity was independent of concentration; therefore, VHH lost its activity in a unimolecular manner, not by aggregation. A VHH mutant lacking Asn, which is susceptible to chemical modifications, had significantly higher heat resistance than did the wild-type protein, indicating the importance of chemical modifications to VHH denaturation. PMID:24739391

  19. Inactivation and reactivation of B. megatherium phage.

    PubMed

    NORTHROP, J H

    1955-11-20

    Preparation of Reversibly Inactivated (R.I.) Phage.- If B. megatherium phage (of any type, or in any stage of purification) is suspended in dilute salt solutions at pH 5-6, it is completely inactivated; i.e., it does not form plaques, or give rise to more phage when mixed with a sensitive organism (Northrop, 1954). The inactivation occurs when the phage is added to the dilute salt solution. If a suspension of the inactive phage in pH 7 peptone is titrated to pH 5 and allowed to stand, the activity gradually returns. The inactivation is therefore reversible. Properties of R.I. Phage.- The R.I. phage is adsorbed by sensitive cells at about the same rate as the active phage. It kills the cells, but no active phage is produced. The R.I. phage therefore has the properties of phage "ghosts" (Herriott, 1951) or of colicines (Gratia, 1925), or phage inactivated by ultraviolet light (Luria, 1947). The R.I. phage is sedimented in the centrifuge at the same rate as active phage. It is therefore about the same size as the active phage. The R.I. phage is most stable in pH 7, 5 per cent peptone, and may be kept in this solution for weeks at 0 degrees C. The rate of digestion of R.I. phage by trypsin, chymotrypsin, or desoxyribonuclease is about the same as that of active phage (Northrop, 1955 a). Effect of Various Substances on the Formation of R.I. Phage.- There is an equilibrium between R.I. phage and active phage. The R.I. form is the stable one in dilute salt solution, pH 5 to 6.5 and at low temperature (<20 degrees C.). At pH >6.5, in dilute salt solution, the R.I. phage changes to the active form. The cycle, active right harpoon over left harpoon inactive phage, may be repeated many times at 0 degrees C. by changing the pH of the solution back and forth between pH 7 and pH 6. Irreversible inactivation is caused by distilled water, some heavy metals, concentrated urea or quanidine solutions, and by l-arginine. Reversible inactivation is prevented by all salts tested (except

  20. Suicide inactivation of catechol 2,3-dioxygenase from Pseudomonas putida mt-2 by 3-halocatechols

    SciTech Connect

    Bartels, I.; Knackmuss, H.J.; Reineke, W.

    1984-03-01

    The inactivation of catechol 2,3-dioxygenase from Pseudomonas putida mt-2 by 3-chloro- and 3-fluorocatechol and the iron-chelating agent Tiron (catechol-3,5-disulfonate) was studied. Whereas inactivation by Tiron is an oxygen-independent and mostly reversible process, inactivation by the 3-halocatechols was only observed in the presence of oxygen and was largely irreversible. The rate constants for inactivation (K/sub 2/) were 1.62 x 10/sup -3/ sec/sup -1/ for 3-chlorocatechol and 2.38 x 10/sup -3/ sec/sup -1/ for 3-fluorocatechol. The inhibitor constants (K/sub i/) were 23 ..mu..M for 3-chlorocatechol and 17 ..mu..M for 3-fluorocatechol. The kinetic data for 3-fluorocatechol could only be obtained in the presence of 2-mercaptoethanol. Besides inactivated enzyme, some 2-hydroxyhexa-2,4-dienoic acid as the actual suicide product of meta-cleavage. A side product of 3-fluorocatechol cleavage is a yellow compound with the spectral characteristics of a 2-hydroxy-6-oxohexa-2,4-dienoci acid indicating 1,6-cleavage. Rates of inactivation by 3-fluorocatechol were reduced in the presence of superoxide dismutase, catalase, formate, and mannitol, which implies that superoxide anion, hydrogen peroxide, and hydroxyl radical exhibit additional inactivation. 64 references.

  1. Enhanced UV inactivation of adenoviruses under polychromatic UV lamps.

    PubMed

    Linden, Karl G; Thurston, Jeanette; Schaefer, Raymond; Malley, James P

    2007-12-01

    Adenovirus is recognized as the most UV-resistant waterborne pathogen of concern to public health microbiologists. The U.S. EPA has stipulated that a UV fluence (dose) of 186 mJ cm(-2) is required for 4-log inactivation credit in water treatment. However, all adenovirus inactivation data to date published in the peer-reviewed literature have been based on UV disinfection experiments using UV irradiation at 253.7 nm produced from a conventional low-pressure UV source. The work reported here presents inactivation data for adenovirus based on polychromatic UV sources and details the significant enhancement in inactivation achieved using these polychromatic sources. When full-spectrum, medium-pressure UV lamps were used, 4-log inactivation of adenovirus type 40 is achieved at a UV fluence of less than 60 mJ cm(-2) and a surface discharge pulsed UV source required a UV fluence of less than 40 mJ cm(-2). The action spectrum for adenovirus type 2 was also developed and partially explains the improved inactivation based on enhancements at wavelengths below 230 nm. Implications for water treatment, public health, and the future of UV regulations for virus disinfection are discussed. PMID:17933932

  2. Initial steps of inactivation at the K+ channel selectivity filter.

    PubMed

    Thomson, Andrew S; Heer, Florian T; Smith, Frank J; Hendron, Eunan; Bernèche, Simon; Rothberg, Brad S

    2014-04-29

    K(+) efflux through K(+) channels can be controlled by C-type inactivation, which is thought to arise from a conformational change near the channel's selectivity filter. Inactivation is modulated by ion binding near the selectivity filter; however, the molecular forces that initiate inactivation remain unclear. We probe these driving forces by electrophysiology and molecular simulation of MthK, a prototypical K(+) channel. Either Mg(2+) or Ca(2+) can reduce K(+) efflux through MthK channels. However, Ca(2+), but not Mg(2+), can enhance entry to the inactivated state. Molecular simulations illustrate that, in the MthK pore, Ca(2+) ions can partially dehydrate, enabling selective accessibility of Ca(2+) to a site at the entry to the selectivity filter. Ca(2+) binding at the site interacts with K(+) ions in the selectivity filter, facilitating a conformational change within the filter and subsequent inactivation. These results support an ionic mechanism that precedes changes in channel conformation to initiate inactivation.

  3. Guinea pig ductus arteriosus. II - Irreversible closure after birth.

    NASA Technical Reports Server (NTRS)

    Fay, F. S.; Cooke, P. H.

    1972-01-01

    To investigate the mechanism underlying irreversibility of ductal closure after birth, studies were undertaken to determine the exact time course for the onset of irreversible closure of the guinea pig ductus arteriosus. Parallel studies of the reactivity of ductal smooth muscle to oxygen and studies of the postpartum cellular changes within the vessel were also carried out.

  4. Conservation-dissipation formalism of irreversible thermodynamics

    NASA Astrophysics Data System (ADS)

    Zhu, Yi; Hong, Liu; Yang, Zaibao; Yong, Wen-An

    2015-06-01

    We propose a conservation-dissipation formalism (CDF) for coarse-grained descriptions of irreversible processes. This formalism is based on a stability criterion for non-equilibrium thermodynamics. The criterion ensures that non-equilibrium states tend to equilibrium in long time. As a systematic methodology, CDF provides a feasible procedure in choosing non-equilibrium state variables and determining their evolution equations. The equations derived in CDF have a unified elegant form. They are globally hyperbolic, allow a convenient definition of weak solutions, and are amenable to existing numerics. More importantly, CDF is a genuinely nonlinear formalism and works for systems far away from equilibrium. With this formalism, we formulate novel thermodynamics theories for heat conduction in rigid bodies and non-isothermal compressible Maxwell fluid flows as two typical examples. In these examples, the non-equilibrium variables are exactly the conjugate variables of the heat fluxes or stress tensors. The new theory generalizes Cattaneo's law or Maxwell's law in a regularized and nonlinear fashion.

  5. Simulations of kinetically irreversible protein aggregate structure.

    PubMed Central

    Patro, S Y; Przybycien, T M

    1994-01-01

    We have simulated the structure of kinetically irreversible protein aggregates in two-dimensional space using a lattice-based Monte-Carlo routine. Our model specifically accounts for the intermolecular interactions between hydrophobic and hydrophilic protein surfaces and a polar solvent. The simulations provide information about the aggregate density, the types of inter-monomer contacts and solvent content within the aggregates, the type and extent of solvent exposed perimeter, and the short- and long-range order all as a function of (i) the extent of monomer hydrophobic surface area and its distribution on the model protein surface and (ii) the magnitude of the hydrophobic-hydrophobic contact energy. An increase in the extent of monomer hydrophobic surface area resulted in increased aggregate densities with concomitant decreased system free energies. These effects are accompanied by increases in the number of hydrophobic-hydrophobic contacts and decreases in the solvent-exposed hydrophobic surface area of the aggregates. Grouping monomer hydrophobic surfaces in a single contiguous stretch resulted in lower aggregate densities and lower short range order. More favorable hydrophobic-hydrophobic contact energies produced structures with higher densities but the number of unfavorable protein-protein contacts was also observed to increase; greater configurational entropy produced the opposite effect. Properties predicted by our model are in good qualitative agreement with available experimental observations. Images FIGURE 6 FIGURE 13 PMID:8061184

  6. Order and disorder in irreversible decay processes.

    PubMed

    Nichols, Jonathan W; Flynn, Shane W; Green, Jason R

    2015-02-14

    Dynamical disorder motivates fluctuating rate coefficients in phenomenological, mass-action rate equations. The reaction order in these rate equations is the fixed exponent controlling the dependence of the rate on the number of species. Here, we clarify the relationship between these notions of (dis)order in irreversible decay, n A → B, n = 1, 2, 3, …, by extending a theoretical measure of fluctuations in the rate coefficient. The measure, Jn-Ln (2)≥0, is the magnitude of the inequality between Jn, the time-integrated square of the rate coefficient multiplied by the time interval of interest, and Ln (2), the square of the time-integrated rate coefficient. Applying the inequality to empirical models for non-exponential relaxation, we demonstrate that it quantifies the cumulative deviation in a rate coefficient from a constant, and so the degree of dynamical disorder. The equality is a bound satisfied by traditional kinetics where a single rate constant is sufficient. For these models, we show how increasing the reaction order can increase or decrease dynamical disorder and how, in either case, the inequality Jn-Ln (2)≥0 can indicate the ability to deduce the reaction order in dynamically disordered kinetics.

  7. Irreversible Electroporation for Colorectal Liver Metastases.

    PubMed

    Scheffer, Hester J; Melenhorst, Marleen C A M; Echenique, Ana M; Nielsen, Karin; van Tilborg, Aukje A J M; van den Bos, Willemien; Vroomen, Laurien G P H; van den Tol, Petrousjka M P; Meijerink, Martijn R

    2015-09-01

    Image-guided tumor ablation techniques have significantly broadened the treatment possibilities for primary and secondary hepatic malignancies. A new ablation technique, irreversible electroporation (IRE), was recently added to the treatment armamentarium. As opposed to thermal ablation, cell death with IRE is primarily induced using electrical energy: electrical pulses disrupt the cellular membrane integrity, resulting in cell death while sparing the extracellular matrix of sensitive structures such as the bile ducts, blood vessels, and bowel wall. The preservation of these structures makes IRE attractive for colorectal liver metastases (CRLM) that are unsuitable for resection and thermal ablation owing to their anatomical location. This review discusses different technical and practical issues of IRE for CRLM: the indications, patient preparations, procedural steps, and different "tricks of the trade" used to improve safety and efficacy of IRE. Imaging characteristics and early efficacy results are presented. Much is still unknown about the exact mechanism of cell death and about factors playing a crucial role in the extent of cell death. At this time, IRE for CRLM should only be reserved for small tumors that are truly unsuitable for resection or thermal ablation because of abutment of the portal triad or the venous pedicles.

  8. Irreversibility in a simple reversible model

    SciTech Connect

    Kumicak, Juraj

    2005-01-01

    This paper studies a parametrized family of familiar generalized Baker maps, viewed as simple models of time-reversible evolution. Mapping the unit square onto itself, the maps are partly contracting and partly expanding, but they preserve the global measure of the definition domain. They possess periodic orbits of any period, and all maps of the set have attractors with well defined structure. The explicit construction of the attractors is described and their structure is studied in detail. There is a precise sense in which one can speak about the absolute age of a state, regardless of whether the latter is applied to a single point, a set of points, or a distribution function. One can then view the whole trajectory as a set of past, present, and future states. This viewpoint is then applied to show that it is impossible to define a priori states with very large 'negative age'. Such states can be defined only a posteriori. This gives precise sense to irreversibility - or the 'arrow of time' - in these time-reversible maps, and is suggested as an explanation of the second law of thermodynamics also for some realistic physical systems.

  9. Recovery of prostacyclin synthesis in vascular smooth muscle cells following self-inactivation and requirement for growth factors

    SciTech Connect

    Bailey, J.M.; Hla, T.T.; Pash, J.M.

    1986-05-01

    The cyclooxygenase enzyme system is a prime example of a metabolic pathway that is regulated by self inactivation. This is believed to occur in part via the irreversible reaction of the endoperoxide intermediate species with the cyclooxygenase enzyme. This inactivation and recovery of activity is similar to the inactivation observed with aspirin which irreversibly acetylates the enzyme. Self inactivation was studied in cultured rat and bovine aorta smooth muscle cells. The production of the prostanoid PGI2 was demonstrated by incubation of a monolayer of cells with 12 ..mu..M C-14 labeled arachidonic acid. Products were analyzed by thin layer chromatography and identified by their comigration with authentic standards and confirmed by gas chromatography/mass spectrometry. Preincubation of the cells for 10 minutes with arachidonic acid at concentrations as low as 1 ..mu..g/mL inactivated the cells to a second challenge with radiolabeled arachidonic acid. Recovery from self inactivation took place over a three hour time period and was similar to the recovery observed with aspirin pretreatment. Recovery was inhibited by addition of 10 ..mu..g/mL cycloheximide to the medium indicating that it involves synthesis of cyclooxygenase protein. Epidermal growth factor was identified as a serum factor responsible for the rapid recovery of cyclooxygenase activity in rat and bovine aorta smooth muscle cells.

  10. Inactivation of Microorganisms

    NASA Astrophysics Data System (ADS)

    Alzamora, Stella Maris; Guerrero, Sandra N.; Schenk, Marcela; Raffellini, Silvia; López-Malo, Aurelio

    Minimal processing techniques for food preservation allow better retention of product flavor, texture, color, and nutrient content than comparable conventional treatments. A wide range of novel alternative physical factors have been intensely investigated in the last two decades. These physical factors can cause inactivation of microorganisms at ambient or sublethal temperatures (e.g., high hydrostatic pressure, pulsed electric fields, ultrasound, pulsed light, and ultraviolet light). These technologies have been reported to reduce microorganism population in foods while avoiding the deleterious effects of severe heating on quality. Among technologies, high-energy ultrasound (i.e., intensities higher than 1 W/cm2, frequencies between 18 and 100 kHz) has attracted considerable interest for food preservation applications (Mason et al., 1996; Povey and Mason, 1998).

  11. Thermal inactivation of microorganisms.

    PubMed

    Smelt, J P P M; Brul, S

    2014-01-01

    This paper serves as an overview of various aspects of thermal processing. Heat processing of foods has a long history and is still one of the most important preservation methods. To guarantee microbiological safety and stability, large safety margins are often applied in traditional heat processes. Because of the need for more fresh like foods, there is a need for milder preservation methods without compromising on safety and stability. The review deals with heat resistance data and mathematical models that describe heat inactivation. The effects of food composition are not yet fully clear and more knowledge of the cell physiology of the target microorganism could be of help in predicting the effects of food constituents. Finally, special attention has been paid to biological time temperature indicators to enable proper process calculations.

  12. Irreversible Electroporation in a Swine Lung Model

    SciTech Connect

    Dupuy, Damian E.; Aswad, Bassam; Ng, Thomas

    2011-04-15

    Purpose: This study was designed to evaluate the safety and tissue effects of IRE in a swine lung model. Methods: This study was approved by the institutional animal care committee. Nine anesthetized domestic swine underwent 15 percutaneous irreversible electroporation (IRE) lesion creations (6 with bipolar and 3 with 3-4 monopolar electrodes) under fluoroscopic guidance and with pancuronium neuromuscular blockade and EKG gating. IRE electrodes were placed into the central and middle third of the right mid and lower lobes in all animals. Postprocedure PA and lateral chest radiographs were obtained to evaluate for pneumothorax. Three animals were sacrificed at 2 weeks and six at 4 weeks. Animals underwent high-resolution CT scanning and PA and lateral radiographs 1 h before sacrifice. The treated lungs were removed en bloc, perfused with formalin, and sectioned. Gross pathologic and microscopic changes after standard hematoxylin and eosin staining were analyzed within the areas of IRE lesion creation. Results: No significant adverse events were identified. CT showed focal areas of spiculated high density ranging in greatest diameter from 1.1-2.2 cm. On gross inspection of the sectioned lung, focal areas of tan discoloration and increased density were palpated in the areas of IRE. Histological analysis revealed focal areas of diffuse alveolar damage with fibrosis and inflammatory infiltration that respected the boundaries of the interlobular septae. No pathological difference could be discerned between the 2- and 4-week time points. The bronchioles and blood vessels within the areas of IRE were intact and did not show signs of tissue injury. Conclusion: IRE creates focal areas of diffuse alveolar damage without creating damage to the bronchioles or blood vessels. Short-term safety in a swine model appears to be satisfactory.

  13. An update on irreversible electroporation of liver tumours.

    PubMed

    Yeung, Enoch S L; Chung, Max W Y; Wong, Keedon; Wong, Clement Y K; So, Enoch C T; Chan, Albert C Y

    2014-08-01

    OBJECTIVE. To investigate the clinical efficacy and safety of irreversible electroporation for ablation of liver tumour in humans. DATA SOURCES. The PubMed and MEDLINE databases were systematically searched. STUDY SELECTION. Clinical research published in English in the last 10 years until October 2013 that address clinical issues related to irreversible electroporation of human liver tumours were selected. "Liver tumor", "local ablative therapy", and "irreversible electroporation" were used as the search terms. DATA EXTRACTION AND SYNTHESIS. The data extracted for this review was analysed by the authors, with a focus on the clinical efficacy and the safety of irreversible electroporation. The complete response rates look promising, ranging from 72% to 100%, except in one study in a subgroup of liver tumours in which the complete response rate was only 50% that was likely due to the inclusion of larger-size tumours. In one study, the local recurrence rate at 12 months was approximately 40%. As for the safety of irreversible electroporation, there were only a few reported complications (cardiac arrhythmia, pneumothorax, and electrolyte disturbance) that were mostly transient and not serious. There was no reported mortality related to the use of irreversible electroporation. CONCLUSION. Irreversible electroporation is a potentially effective liver tumour ablative therapy that gives rise to only mild and transient side-effects. Further studies with better patient selection criteria and longer follow-up are needed to clarify its role as a first-line liver tumour treatment modality.

  14. Pilot study of irreversible electroporation for intracranial surgery.

    PubMed

    Garcia, Paulo A; Rossmeisl, John H; Robertson, John; Ellis, Thomas L; Davalos, Rafael V

    2009-01-01

    Irreversible electroporation (IRE) is a new minimally invasive technique to treat cancer using intense but short electric pulses. This technique is unique because of its non-thermal mechanism of tissue ablation. Furthermore it can be predicted with numerical models and can be confirmed with ultrasound and MRI. We present some preliminary results on the safety of using irreversible electroporation for canine brain surgery. We also present the electric field (460 V/cm - 560 V/cm) necessary for focal ablation of canine brain tissue and provide some guidelines for treatment planning and execution. This preliminary study is the first step towards using irreversible electroporation as a brain cancer treatment. PMID:19964170

  15. Complete inactivation of HIV-1 using photo-labeled non-nucleoside reverse transcriptase inhibitors.

    PubMed

    Rios, Adan; Quesada, Jorge; Anderson, Dallas; Goldstein, Allan; Fossum, Theresa; Colby-Germinario, Susan; Wainberg, Mark A

    2011-01-01

    We demonstrate that a photo-labeled derivative of the non-nucleoside reverse transcriptase inhibitor (NNRTI) dapivirine termed DAPY, when used together with exposure to ultraviolet light, was able to completely and irreversibly inactivate both HIV-1 RT activity as well as infectiousness in each of a T cell line and peripheral blood mononuclear cells. Control experiments using various concentrations of DAPY revealed that a combination of exposure to ultraviolet light together with use of the specific, high affinity photo-labeled compound was necessary for complete inactivation to occur. This method of HIV RT inactivation may have applicability toward preservation of an intact viral structure and warrants further investigation in regard to the potential of this approach to elicit a durable, broad protective immune response. PMID:20937333

  16. Fourteen. beta. -(bromoacetamido)morphine irreversibly labels. mu. opioid receptors in rat brain membranes

    SciTech Connect

    Bidlack, J.M.; Frey, D.K.; Seyed-Mozaffari, A.; Archer, S. )

    1989-05-16

    The binding properties of 14{beta}-(bromoacetamido)morphine (BAM) and the ability of BAM to irreversibly inhibit opioid binding to rat brain membranes were examined to characterize the affinity and selectivity of BAM as an irreversible affinity ligand for opioid receptors. BAM had the same receptor selectivity as morphine, with a 3-5-fold decrease in affinity for the different types of opioid receptors. When brain membranes were incubated with BAM, followed by extensive washing, opioid binding was restored to control levels. However, when membranes were incubated with dithiothreitol (DTT), followed by BAM, and subsequently washed, 90% of the 0.25 nM ({sup 3}H)(D-Ala{sup 2},(Me)Phe{sup 4},Gly(ol){sup 5})enkephalin (DAGO) binding was irreversibly inhibited as a result of the specific alkylation of a sulfhydryl group at the {mu} binding site. This inhibition was dependent on the concentrations of both DTT and BAM. The {mu} receptor specificity of BAM alkylation was demonstrated by the ability of BAM alkylated membranes to still bind the {delta}-selective peptide ({sup 3}H)(D-penicillamine{sup 2},D-penicillamine{sup 5})enkephalin (DPDPE) and (-)-({sup 3}H)bremazocine in the presence of {mu} and {delta} blockers, selective for {kappa} binding sites. Morphine and naloxone partially protected the binding site from alkylation with BAM, while ligands that did not bind to the {mu}s site did not afford protection. These studies have demonstrated that when a disulfide bond at or near {mu} opioid binding sites was reduced, BAM could then alkylate this site, resulting in the specific irreversible labeling of {mu} opioid receptors.

  17. Microscopic reversibility and macroscopic irreversibility: A lattice gas model

    NASA Astrophysics Data System (ADS)

    Pérez-Cárdenas, Fernando C.; Resca, Lorenzo; Pegg, Ian L.

    2016-09-01

    We present coarse-grained descriptions and computations of the time evolution of a lattice gas system of indistinguishable particles, whose microscopic laws of motion are exactly reversible, in order to investigate how or what kind of macroscopically irreversible behavior may eventually arise. With increasing coarse-graining and number of particles, relative fluctuations of entropy rapidly decrease and apparently irreversible behavior unfolds. Although that behavior becomes typical in those limits and within a certain range, it is never absolutely irreversible for any individual system with specific initial conditions. Irreversible behavior may arise in various ways. We illustrate one possibility by replacing detailed integer occupation numbers at lattice sites with particle probability densities that evolve diffusively.

  18. Dissipation and irreversibility for models of mechanochemical machines

    NASA Astrophysics Data System (ADS)

    Brown, Aidan; Sivak, David

    For biological systems to maintain order and achieve directed progress, they must overcome fluctuations so that reactions and processes proceed forwards more than they go in reverse. It is well known that some free energy dissipation is required to achieve irreversible forward progress, but the quantitative relationship between irreversibility and free energy dissipation is not well understood. Previous studies focused on either abstract calculations or detailed simulations that are difficult to generalize. We present results for mechanochemical models of molecular machines, exploring a range of model characteristics and behaviours. Our results describe how irreversibility and dissipation trade off in various situations, and how this trade-off can depend on details of the model. The irreversibility-dissipation trade-off points towards general principles of microscopic machine operation or process design. Our analysis identifies system parameters which can be controlled to bring performance to the Pareto frontier.

  19. Irreversibility of financial time series: A graph-theoretical approach

    NASA Astrophysics Data System (ADS)

    Flanagan, Ryan; Lacasa, Lucas

    2016-04-01

    The relation between time series irreversibility and entropy production has been recently investigated in thermodynamic systems operating away from equilibrium. In this work we explore this concept in the context of financial time series. We make use of visibility algorithms to quantify, in graph-theoretical terms, time irreversibility of 35 financial indices evolving over the period 1998-2012. We show that this metric is complementary to standard measures based on volatility and exploit it to both classify periods of financial stress and to rank companies accordingly. We then validate this approach by finding that a projection in principal components space of financial years, based on time irreversibility features, clusters together periods of financial stress from stable periods. Relations between irreversibility, efficiency and predictability are briefly discussed.

  20. CHLORINE INACTIVATION OF BACILLUS ENDOSPORES

    EPA Science Inventory

    The possibility of a bioterrorism event resulting in the release of Bacillus anthracis endospores into a drinking water distribution system necessitates research into means by which these endospores can be inactivated. This study was designed to determine the chlorine resistance...

  1. Influenza Vaccine, Inactivated or Recombinant

    MedlinePlus

    ... die from flu, and many more are hospitalized.Flu vaccine can:keep you from getting flu, make flu ... inactivated or recombinant influenza vaccine?A dose of flu vaccine is recommended every flu season. Children 6 months ...

  2. Extended irreversible thermodynamics and the quality of temperature and pressure

    NASA Astrophysics Data System (ADS)

    Bhalekar, Anil A.

    1999-08-01

    It is reiterated that without a Gibbs-Duhem equation no thermodynamic description ofirreversible and reversible processes exists. It is shown with the help of Gibbs-Duhem equation of extended irreversible thermodynamics that the physical contents of intensive quantities, the temperature and the pressure, do not change in going from reversible to irreversible processes. This confirms well with the earlier demonstrations of Eu and Garcia-Colin.

  3. Anisotropic shift of the irreversibility line by neutron irradiation

    SciTech Connect

    Sauerzopf, F.M.; Wiesinger, H.P.; Weber, H.W. ); Crabtree, G.W.; Frischherz, M.C.; Kirk, M.A. )

    1991-09-01

    The irreversibility line of high-{Tc} superconductors is shifted considerably by irradiating the material with fast neutrons. The anisotropic and non-monotonous shift is qualitatively explained by a simple model based on an interaction between three pinning mechanisms, the intrinsic pinning by the ab-planes, the weak pinning by the pre-irradiation defect structure, and strong pinning by neutron induced defect cascades. A correlation between the cascade density and the position of the irreversibility line is observed.

  4. Irreversible pulpitis and achieving profound anesthesia: Complexities and managements

    PubMed Central

    Modaresi, Jalil; Davoudi, Amin; Badrian, Hamid; Sabzian, Roya

    2016-01-01

    Dental pain management is one of the most critical aspects of modern dentistry. Irreversible pulpitis and further root canal therapy might cause an untolerated pain to the patients. The improvements in anesthetic agents and techniques were one of the advantages of studying nerve biology and stimulation. This article tried to overview of the nerve activities in inflammatory environments or induced pain. Furthermore, the proper advises, and supplementary techniques were reviewed for better pain management of irreversible pulpitis. PMID:26957681

  5. Calcium-mediated inactivation of the calcium conductance in cesium-loaded frog heart cells.

    PubMed

    Mentrard, D; Vassort, G; Fischmeister, R

    1984-01-01

    Ca current inactivation was investigated in frog atrial muscle under voltage-clamp conditions. To inhibit the outward currents, experiments were performed on Cs-loaded fibers and in 20 mM Cs (K-free) Ringer with 4-AP added. Inactivation, produced by a conditioning pulse, was measured by reducing the current during a subsequent test pulse. The extent of inactivation increased initially with prepulse amplitude and then decreased as the prepulse potential became progressively positive. Relative inactivation follows a U-shaped curve. When Sr was substituted for Ca, both the degree and the rate of inactivation decreased. Relative inactivation appeared to be linearly related to the amount of divalent cations (Ca and Sr) carried into the cell during the prepulse. Elevating Ca enhanced peak current and accelerated its decline. Elevating Mg decreased peak current and slowed its decline. An application of Na-free (LiCl) solution resulted in a somewhat smaller but faster inactivating current. Adrenaline increased and D600 decreased the maximal Ca conductance with little alteration in the inactivation rate; Co decreased both peak current and the rate of inactivation. Enhancement of the outward currents, reduced driving force, and intracellular surface charge screening do not adequately account for the above results. Evidence was considered that Ca entry mediates most of Ca current inactivation in frog atrial fibers. Removal from inactivation was also investigated in normal-Ca, Ca-rich, and Sr solutions. Recovery after partial inactivation by high depolarization was biphasic. Recovery was slowed by 10 Ca and accelerated by 1.8 Sr, whereas opposite effects have been shown on activation.

  6. Human PIEZO1: removing inactivation.

    PubMed

    Bae, Chilman; Gottlieb, Philip A; Sachs, Frederick

    2013-08-20

    PIEZO1 is an inactivating eukaryotic cation-selective mechanosensitive ion channel. Two sites have been located in the channel that when individually mutated lead to xerocytotic anemia by slowing inactivation. By introducing mutations at two sites, one associated with xerocytosis and the other artificial, we were able to remove inactivation. The double mutant (DhPIEZO1) has a substitution of arginine for methionine (M2225R) and lysine for arginine (R2456K). The loss of inactivation was accompanied by ∼30-mmHg shift of the activation curve to lower pressures and slower rates of deactivation. The slope sensitivity of gating was the same for wild-type and mutants, indicating that the dimensional changes between the closed and open state are unaffected by the mutations. The unitary channel conductance was unchanged by mutations, so these sites are not associated with pore. DhPIEZO1 was reversibly inhibited by the peptide GsMTx4 that acted as a gating modifier. The channel kinetics were solved using complex stimulus waveforms and the data fit to a three-state loop in detailed balance. The reaction had two pressure-dependent rates, closed to open and inactivated to closed. Pressure sensitivity of the opening rate with no sensitivity of the closing rate means that the energy barrier between them is located near the open state. Mutant cycle analysis of inactivation showed that the two sites interacted strongly, even though they are postulated to be on opposite sides of the membrane. PMID:23972840

  7. Mitochondrial aconitase reaction with nitric oxide, S-nitrosoglutathione, and peroxynitrite: mechanisms and relative contributions to aconitase inactivation.

    PubMed

    Tórtora, Verónica; Quijano, Celia; Freeman, Bruce; Radi, Rafael; Castro, Laura

    2007-04-01

    Using highly purified recombinant mitochondrial aconitase, we determined the kinetics and mechanisms of inactivation mediated by nitric oxide (*NO), nitrosoglutathione (GSNO), and peroxynitrite (ONOO(-)). High *NO concentrations are required to inhibit resting aconitase. Brief *NO exposures led to a reversible inhibition competitive with isocitrate (K(I)=35 microM). Subsequently, an irreversible inactivation (0.65 M(-1) s(-1)) was observed. Irreversible inactivation was mediated by GSNO also, both in the absence and in the presence of substrates (0.23 M(-1) s(-1)). Peroxynitrite reacted with the [4Fe-4S] cluster, yielding the inactive [3Fe-4S] enzyme (1.1 x 10(5) M(-1) s(-1)). Carbon dioxide enhanced ONOO(-)-dependent inactivation via reaction of CO(3)*(-) with the [4Fe-4S] cluster (3 x 10(8) M(-1) s(-1)). Peroxynitrite also induced m-aconitase tyrosine nitration but this reaction did not contribute to enzyme inactivation. Computational modeling of aconitase inactivation by O(2)*(-) and *NO revealed that, when NO is produced and readily consumed, measuring the amount of active aconitase remains a sensitive method to detect variations in O(2)*(-) production in cells but, when cells are exposed to high concentrations of NO, aconitase inactivation does not exclusively reflect changes in rates of O(2)*(-) production. In the latter case, extents of aconitase inactivation reflect the formation of secondary reactive species, specifically ONOO(-) and CO(3)*(-), which also mediate m-aconitase tyrosine nitration, a footprint of reactive *NO-derived species.

  8. Inactivation of mitochondrial ATPase by ultraviolet light

    SciTech Connect

    Chavez, E.; Cuellar, A.

    1984-05-01

    The present work describes experiments that show that far-ultraviolet irradiation induce the inhibition of ATPase activity in both membrane-bound and soluble F1. It was also found that ultraviolet light promotes the release of tightly bound adenine nucleotides from F1-ATPase. Experiments carried out with submitochondrial particles indicate that succinate partially protects against these effects of ultraviolet light. Titration of sulfhydryl groups in both irradiated submitochondrial particles and soluble F1-ATPase indicates that a conformational change induced by photochemical modifications of amino acid residues appears involved in the inactivation of the enzyme. Finally, experiments are described which show that the tyrosine residue located in the active site of F1-ATPase is modified by ultraviolet irradiation.

  9. Attribution of irreversible loss to anthropogenic climate change

    NASA Astrophysics Data System (ADS)

    Huggel, Christian; Bresch, David; Hansen, Gerrit; James, Rachel; Mechler, Reinhard; Stone, Dáithí; Wallimann-Helmer, Ivo

    2016-04-01

    The Paris Agreement (2015) under the UNFCCC has anchored loss and damage in a separate article which specifies that understanding and support should be enhanced in areas addressing loss and damage such as early warning, preparedness, insurance and resilience. Irreversible loss is a special category under loss and damage but there is still missing clarity over what irreversible loss actually includes. Many negative impacts of climate change may be handled or mitigated by existing risk management, reduction and absorption approaches. Irreversible loss, however, is thought to be insufficiently addressed by risk management. Therefore, countries potentially or actually affected by irreversible loss are calling for other measures such as compensation, which however is highly contested in international climate policy. In Paris (2015) a decision was adopted that loss and damage as defined in the respective article of the agreement does not involve compensation and liability. Nevertheless, it is likely that some sort of mechanism will eventually need to come into play for irreversible loss due to anthropogenic climate change, which might involve compensation, other forms of non-monetary reparation, or transformation. Furthermore, climate litigation has increasingly been attempted to address negative effects of climate change. In this context, attribution is important to understand the drivers of change, what counts as irreversible loss due to climate change, and, possibly, who or what is responsible. Here we approach this issue by applying a detection and attribution perspective on irreversible loss. We first analyze detected climate change impacts as assessed in the IPCC Fifth Assessment Report. We distinguish between irreversible loss in physical, biological and human systems, and accordingly identify the following candidates of irreversible loss in these systems: loss of glaciers and ice sheets, loss of subsurface ice (permafrost) and related loss of lake systems; loss

  10. Ribosome-inactivating proteins: from plant defense to tumor attack.

    PubMed

    de Virgilio, Maddalena; Lombardi, Alessio; Caliandro, Rocco; Fabbrini, Maria Serena

    2010-11-01

    Ribosome-inactivating proteins (RIPs) are EC3.2.32.22 N-glycosidases that recognize a universally conserved stem-loop structure in 23S/25S/28S rRNA, depurinating a single adenine (A4324 in rat) and irreversibly blocking protein translation, leading finally to cell death of intoxicated mammalian cells. Ricin, the plant RIP prototype that comprises a catalytic A subunit linked to a galactose-binding lectin B subunit to allow cell surface binding and toxin entry in most mammalian cells, shows a potency in the picomolar range. The most promising way to exploit plant RIPs as weapons against cancer cells is either by designing molecules in which the toxic domains are linked to selective tumor targeting domains or directly delivered as suicide genes for cancer gene therapy. Here, we will provide a comprehensive picture of plant RIPs and discuss successful designs and features of chimeric molecules having therapeutic potential. PMID:22069572

  11. Irreversible climate change due to carbon dioxide emissions.

    PubMed

    Solomon, Susan; Plattner, Gian-Kasper; Knutti, Reto; Friedlingstein, Pierre

    2009-02-10

    The severity of damaging human-induced climate change depends not only on the magnitude of the change but also on the potential for irreversibility. This paper shows that the climate change that takes place due to increases in carbon dioxide concentration is largely irreversible for 1,000 years after emissions stop. Following cessation of emissions, removal of atmospheric carbon dioxide decreases radiative forcing, but is largely compensated by slower loss of heat to the ocean, so that atmospheric temperatures do not drop significantly for at least 1,000 years. Among illustrative irreversible impacts that should be expected if atmospheric carbon dioxide concentrations increase from current levels near 385 parts per million by volume (ppmv) to a peak of 450-600 ppmv over the coming century are irreversible dry-season rainfall reductions in several regions comparable to those of the "dust bowl" era and inexorable sea level rise. Thermal expansion of the warming ocean provides a conservative lower limit to irreversible global average sea level rise of at least 0.4-1.0 m if 21st century CO(2) concentrations exceed 600 ppmv and 0.6-1.9 m for peak CO(2) concentrations exceeding approximately 1,000 ppmv. Additional contributions from glaciers and ice sheet contributions to future sea level rise are uncertain but may equal or exceed several meters over the next millennium or longer.

  12. Irreversible climate change due to carbon dioxide emissions.

    PubMed

    Solomon, Susan; Plattner, Gian-Kasper; Knutti, Reto; Friedlingstein, Pierre

    2009-02-10

    The severity of damaging human-induced climate change depends not only on the magnitude of the change but also on the potential for irreversibility. This paper shows that the climate change that takes place due to increases in carbon dioxide concentration is largely irreversible for 1,000 years after emissions stop. Following cessation of emissions, removal of atmospheric carbon dioxide decreases radiative forcing, but is largely compensated by slower loss of heat to the ocean, so that atmospheric temperatures do not drop significantly for at least 1,000 years. Among illustrative irreversible impacts that should be expected if atmospheric carbon dioxide concentrations increase from current levels near 385 parts per million by volume (ppmv) to a peak of 450-600 ppmv over the coming century are irreversible dry-season rainfall reductions in several regions comparable to those of the "dust bowl" era and inexorable sea level rise. Thermal expansion of the warming ocean provides a conservative lower limit to irreversible global average sea level rise of at least 0.4-1.0 m if 21st century CO(2) concentrations exceed 600 ppmv and 0.6-1.9 m for peak CO(2) concentrations exceeding approximately 1,000 ppmv. Additional contributions from glaciers and ice sheet contributions to future sea level rise are uncertain but may equal or exceed several meters over the next millennium or longer. PMID:19179281

  13. Heat inactivation kinetics of Hypocrea orientalis β-glucosidase with enhanced thermal stability by glucose.

    PubMed

    Xu, Xin-Qi; Shi, Yan; Wu, Xiao-Bing; Zhan, Xi-Lan; Zhou, Han-Tao; Chen, Qing-Xi

    2015-11-01

    Thermal inactivation kinetics of Hypocrea orientalis β-glucosidase and effect of glucose on thermostability of the enzyme have been determined in this paper. Kinetic studies showed that the thermal inactivation was irreversible and first-order reaction. The microscopic rate constants for inactivation of free enzyme and substrate-enzyme complex were both determined, which suggested that substrates can protect β-glucosidase against thermal deactivation effectively. On the other hand, glucose was found to protect β-glucosidase from heat inactivation to remain almost whole activity below 70°C at 20mM concentration, whereas the apparent inactivation rate of BG decreased to be 0.3×10(-3)s(-1) in the presence of 5mM glucose, smaller than that of sugar-free enzyme (1.91×10(-3)s(-1)). The intrinsic fluorescence spectra results showed that glucose also had stabilizing effect on the conformation of BG against thermal denaturation. Docking simulation depicted the interaction mode between glucose and active residues of the enzyme to produce stabilizing effect.

  14. Activation of a muscarinic receptor selectively inhibits a rapidly inactivated Ca2+ current in rat sympathetic neurons.

    PubMed Central

    Wanke, E; Ferroni, A; Malgaroli, A; Ambrosini, A; Pozzan, T; Meldolesi, J

    1987-01-01

    Sympathetic neurons dissociated from the superior cervical ganglion of 2-day-old rats were studied by whole-cell patch clamp and by fura-2 measurements of the cytosolic free Ca2+ concentration, [Ca2+]i. Step depolarizations in the presence of tetrodotoxin and hexamethonium triggered two Ca2+ currents that differed in the voltage dependence of activation and kinetics of inactivation. These currents resemble the L and N currents previously described in chicken sensory neurons [Nowycky, M. C., Fox, A. P. & Tsien, R. W. (1985) Nature (London) 316, 440-442]. Treatment with acetylcholine resulted in the rapid (within seconds), selective, and reversible inhibition of the rapidly inactivated, N-type current, whereas the long-lasting L-type current remained unaffected. The high sensitivity to blocker drugs (atropine, pirenzepine) indicated that this effect of acetylcholine was due to a muscarinic M1 receptor. Intracellular perfusion with nonhydrolyzable guanine nucleotide analogs or pretreatment of the neurons with pertussis toxin had profound effects on the Ca2+ current modulation. Guanosine 5'-[gamma-thio]triphosphate caused the disappearance of the N-type current (an effect akin to that of acetylcholine, but irreversible), whereas guanosine 5'-[beta-thio]diphosphate and pertussis toxin pretreatment prevented the acetylcholine-induced inhibition. In contrast, cAMP, applied intracellularly together with 3-isobutyl-1-methylxanthine, as well as activators and inhibitors of protein kinase C, were without effect. Acetylcholine caused shortening of action potentials in neurons treated with tetraethylammonium to partially block K+ channels. Moreover, when applied to neurons loaded with the fluorescent indicator fura-2, acetylcholine failed to appreciably modify [Ca2+]i at rest but caused a partial blunting of the initial [Ca2+]i peak induced by depolarization with high K+. This effect was blocked by muscarinic antagonists and pertussis toxin and was unaffected by protein kinase

  15. Mechanisms of endospore inactivation under high pressure.

    PubMed

    Reineke, Kai; Mathys, Alexander; Heinz, Volker; Knorr, Dietrich

    2013-06-01

    It is well known that spore germination and inactivation can be achieved within a broad temperature and pressure range. The existing literature, however, reports contradictory results concerning the effectiveness of different pressure-temperature combinations and the underlying inactivation mechanism(s). Much of the published kinetic data are prone to error as a result of unstable process conditions or an incomplete investigation of the entire inactivation pathway. Here, we review this field of research, and also discuss an inactivation mechanism of at least two steps and propose an inactivation model based on current data. Further, spore resistance properties and matrix interactions are linked to spore inactivation effectiveness.

  16. Epilepsy (partial)

    PubMed Central

    2011-01-01

    Introduction About 3% of people will be diagnosed with epilepsy during their lifetime, but about 70% of people with epilepsy eventually go into remission. Methods and outcomes We conducted a systematic review and aimed to answer the following clinical questions: What are the effects of starting antiepileptic drug treatment following a single seizure? What are the effects of drug monotherapy in people with partial epilepsy? What are the effects of additional drug treatments in people with drug-resistant partial epilepsy? What is the risk of relapse in people in remission when withdrawing antiepileptic drugs? What are the effects of behavioural and psychological treatments for people with epilepsy? What are the effects of surgery in people with drug-resistant temporal lobe epilepsy? We searched: Medline, Embase, The Cochrane Library, and other important databases up to July 2009 (Clinical Evidence reviews are updated periodically; please check our website for the most up-to-date version of this review). We included harms alerts from relevant organisations such as the US Food and Drug Administration (FDA) and the UK Medicines and Healthcare products Regulatory Agency (MHRA). Results We found 83 systematic reviews, RCTs, or observational studies that met our inclusion criteria. We performed a GRADE evaluation of the quality of evidence for interventions. Conclusions In this systematic review we present information relating to the effectiveness and safety of the following interventions: antiepileptic drugs after a single seizure; monotherapy for partial epilepsy using carbamazepine, gabapentin, lamotrigine, levetiracetam, phenobarbital, phenytoin, sodium valproate, or topiramate; addition of second-line drugs for drug-resistant partial epilepsy (allopurinol, eslicarbazepine, gabapentin, lacosamide, lamotrigine, levetiracetam, losigamone, oxcarbazepine, retigabine, tiagabine, topiramate, vigabatrin, or zonisamide); antiepileptic drug withdrawal for people with partial or

  17. Spectral line intensity irreversibility in circulatory plasma magnetization processes

    SciTech Connect

    Qu, Z. Q.; Dun, G. T.

    2012-01-23

    Spectral line intensity variation is found to be irreversible in circulatory plasma magnetization process by experiments described in this paper, i.e., the curves illustrating spectral line photon fluxes irradiated from a light source immerged in a magnetic field by increasing the magnetic induction cannot be reproduced by decreasing the magnetic induction within the errors. There are two plasma magnetization patterns found. One shows that the intensities are greater at the same magnetic inductions during the magnetic induction decreasing process after the increasing, and the other gives the opposite effect. This reveals that the magneto-induced excitation and de-excitation process is irreversible like ferromagnetic magnetization. But the two irreversible processes are very different in many aspects stated in the text.

  18. Irreversible aggregation of flexible chainlike walkers without adherence.

    PubMed

    Mashiko, Takashi

    2008-07-01

    The flexible chainlike walker (FCW) model is proposed as a minimal model of a deformable moving object and as an extension of the regular random-walk model. The many-body system of FCWs is studied by numerical simulations on a square lattice. It is shown that FCWs aggregate spontaneously and irreversibly where no adherence is assumed, in contrast to the established aggregation models, where adherence is indispensable for their occurrence and irreversibility. This type of aggregation is enabled by and demonstrates the significance of the deformability of moving objects.

  19. Entanglement irreversibility from quantum discord and quantum deficit.

    PubMed

    Cornelio, Marcio F; de Oliveira, Marcos C; Fanchini, Felipe F

    2011-07-01

    We relate the problem of irreversibility of entanglement with the recently defined measures of quantum correlation--quantum discord and one-way quantum deficit. We show that the entanglement of formation is always strictly larger than the coherent information and the entanglement cost is also larger in most cases. We prove irreversibility of entanglement under local operations and classical communication for a family of entangled states. This family is a generalization of the maximally correlated states for which we also give an analytic expression for the distillable entanglement, the relative entropy of entanglement, the distillable secret key, and the quantum discord.

  20. First Law of Thermodynamics; Irreversible and Reversible Processes

    NASA Astrophysics Data System (ADS)

    Craig, Norman C.; Gislason, Eric A.

    2002-02-01

    The experimental basis for the first law of thermodynamics and related operational definitions are reviewed. An alternative, adiabatic-work formulation of the first law is evaluated. New mathematical expressions for irreversible work in isothermal and adiabatic expansions and compressions of an ideal gas are presented. The work for these irreversible processes is compared graphically with the reversible limit. For the adiabatic case some numerical explorations are also done. An operational definition for reversible processes in purely first-law terms is presented and applied.

  1. A new method for making casts from irreversible hydrocolloid impressions.

    PubMed

    Steas, A

    1991-03-01

    This method of making casts from alginate (irreversible hydrocolloid) edentulous impressions is a departure from the usual procedures. It consists of recognizing the weaknesses of the materials being used and handling them to the best advantage to minimize deleterious effects. A proper dental stone mixed with an accelerator is painted over the entire anatomic surface of the impression. A base is added only after the first application of stone sets. This method protects against volume change of the irreversible hydrocolloid impression material and distortion of the unsupported portions of the impression.

  2. A dielectric barrier discharge terminally inactivates RNase A by oxidizing sulfur-containing amino acids and breaking structural disulfide bonds

    NASA Astrophysics Data System (ADS)

    Lackmann, J.-W.; Baldus, S.; Steinborn, E.; Edengeiser, E.; Kogelheide, F.; Langklotz, S.; Schneider, S.; Leichert, L. I. O.; Benedikt, J.; Awakowicz, P.; Bandow, J. E.

    2015-12-01

    RNases are among the most stable proteins in nature. They even refold spontaneously after heat inactivation, regaining full activity. Due to their stability and universal presence, they often pose a problem when experimenting with RNA. We investigated the capabilities of nonthermal atmospheric-pressure plasmas to inactivate RNase A and studied the inactivation mechanism on a molecular level. While prolonged heating above 90 °C is required for heat inactivating RNase A, direct plasma treatment with a dielectric barrier discharge (DBD) source caused permanent inactivation within minutes. Circular dichroism spectroscopy showed that DBD-treated RNase A unfolds rapidly. Raman spectroscopy indicated methionine modifications and formation of sulfonic acid. A mass spectrometry-based analysis of the protein modifications that occur during plasma treatment over time revealed that methionine sulfoxide formation coincides with protein inactivation. Chemical reduction of methionine sulfoxides partially restored RNase A activity confirming that sulfoxidation is causal and sufficient for RNase A inactivation. Continued plasma exposure led to over-oxidation of structural disulfide bonds. Using antibodies, disulfide bond over-oxidation was shown to be a general protein inactivation mechanism of the DBD. The antibody’s heavy and light chains linked by disulfide bonds dissociated after plasma exposure. Based on their ability to inactivate proteins by oxidation of sulfur-containing amino acids and over-oxidation of disulfide bonds, DBD devices present a viable option for inactivating undesired or hazardous proteins on heat or solvent-sensitive surfaces.

  3. Inactivation of enveloped virus by laser-driven protein aggregation

    NASA Astrophysics Data System (ADS)

    Tsen, Shaw-Wei D.; Chapa, Travis; Beatty, Wandy; Tsen, Kong-Thon; Yu, Dong; Achilefu, Samuel

    2012-12-01

    Ultrafast lasers in the visible and near-infrared range have emerged as a potential new method for pathogen reduction of blood products and pharmaceuticals. However, the mechanism of enveloped virus inactivation by this method is unknown. We report the inactivation as well as the molecular and structural effects caused by visible (425 nm) femtosecond laser irradiation on murine cytomegalovirus (MCMV), an enveloped, double-stranded DNA virus. Our results show that laser irradiation (1) caused a 5-log reduction in MCMV titer, (2) did not cause significant changes to the global structure of MCMV virions including membrane and capsid, as assessed by electron microscopy, (3) produced no evidence of double-strand breaks or crosslinking in MCMV genomic DNA, and (4) caused selective aggregation of viral capsid and tegument proteins. We propose a model in which ultrafast laser irradiation induces partial unfolding of viral proteins by disrupting hydrogen bonds and/or hydrophobic interactions, leading to aggregation of closely associated viral proteins and inactivation of the virus. These results provide new insight into the inactivation of enveloped viruses by visible femtosecond lasers at the molecular level, and help pave the way for the development of a new ultrafast laser technology for pathogen reduction.

  4. Thiocyanate and hydroxyl ions inactivate the scrapie agent.

    PubMed Central

    Prusiner, S B; Groth, D F; McKinley, M P; Cochran, S P; Bowman, K A; Kasper, K C

    1981-01-01

    To probe the macromolecular structure of the scrapie agent and explore conditions for monomerization, the stability of the agent in low concentrations of inorganic ions was determined. A reduction by a factor of 10(5) in scrapie titer was found on exposure of the agent to 1 M KSCN or 0.3 M NaOH. In addition to the inactivation by thiocyanate ions, other chaotropic ions such as guanidinium and trichloroacetate inactivate the scrapie agent. Removal of thiocyanate ions by dialysis or glass permeation chromatography prevented the reduction in scrapie agent infectivity. Addition of equimolar amounts of (NH4)2SO4, a nonchaotrope, to preparations containing 1 M KSCN also prevented the loss of scrapie infectivity. In contrast, neutralization of the alkali-treated fractions with HCl did not restore infectivity. Acidification of partially purified fractions did not cause inactivation of the agent but did result in precipitation of the infectious agent. Inactivation by relatively low concentrations of chaotropic ions is consistent with many observations, all of which suggest that the scrapie agent contains a protein component that is essential for the maintenance of infectivity. Thus, it is unlikely that the agent is composed only of a "naked" nucleic acid. Certainly, if the agent were a naked nucleic acid, its lability in alkali virtually eliminates the possibility that it is composed of a single-stranded molecule of DNA. PMID:6794034

  5. Inactivation of enveloped virus by laser-driven protein aggregation.

    PubMed

    Tsen, Shaw-Wei D; Chapa, Travis; Beatty, Wandy; Tsen, Kong-Thon; Yu, Dong; Achilefu, Samuel

    2012-12-01

    Ultrafast lasers in the visible and near-infrared range have emerged as a potential new method for pathogen reduction of blood products and pharmaceuticals. However, the mechanism of enveloped virus inactivation by this method is unknown. We report the inactivation as well as the molecular and structural effects caused by visible (425 nm) femtosecond laser irradiation on murine cytomegalovirus (MCMV), an enveloped, double-stranded DNA virus. Our results show that laser irradiation (1) caused a 5-log reduction in MCMV titer, (2) did not cause significant changes to the global structure of MCMV virions including membrane and capsid, as assessed by electron microscopy, (3) produced no evidence of double-strand breaks or crosslinking in MCMV genomic DNA, and (4) caused selective aggregation of viral capsid and tegument proteins. We propose a model in which ultrafast laser irradiation induces partial unfolding of viral proteins by disrupting hydrogen bonds and/or hydrophobic interactions, leading to aggregation of closely associated viral proteins and inactivation of the virus. These results provide new insight into the inactivation of enveloped viruses by visible femtosecond lasers at the molecular level, and help pave the way for the development of a new ultrafast laser technology for pathogen reduction. PMID:23224114

  6. INACTIVATION OF CRYPTOSPORIDIUM PARVUM OOCYSTS WITH OZONE

    EPA Science Inventory

    Ozone inactivation rates for Cryptosporidium parvum (C. parvum) oocysts were determined with an in-vitro excystation method based on excysted sporozoite counts. Results were consistent with published animal infectivity data for the same C. parvum strain. The inactivation kinetics...

  7. CRYPTOSPORIDIUM LOG INACTIVATION CALCULATION METHODS

    EPA Science Inventory

    Appendix O of the Surface Water Treatment Rule (SWTR) Guidance Manual introduces the CeffT10 (i.e., reaction zone outlet C value and T10 time) method for calculating ozone CT value and Giardia and virus log inactivation. The LT2ESWTR Pre-proposal Draft Regulatory Language for St...

  8. CRYPTOSPORIDIUM INACTIVATION AND REMOVAL RESEARCH

    EPA Science Inventory

    Bench- and pilot-scale tests were performed to assess the ability of conventional treatment, ozonation and chlorine dioxide to remove and inactivate Cryptosporidium oocysts. The impacts of coagulant type, coagulant dose, raw water quality, filter loading rates and filter media w...

  9. The Irreversible Process of University "Democratization": The Danish Case.

    ERIC Educational Resources Information Center

    Pedersen, Mogens N.

    1988-01-01

    The Danish experience with university democratization suggests that the process is irreversible and that its progress is determined by how the initial change was begun two decades ago. It is also proposed that government attempts to intervene and revoke traditional institutional autonomy threaten to invalidate the progress made. (Author/MSE)

  10. When an Adiabatic Irreversible Expansion or Compression Becomes Reversible

    ERIC Educational Resources Information Center

    Anacleto, Joaquim; Ferreira, J. M.; Soares, A. A.

    2009-01-01

    This paper aims to contribute to a better understanding of the concepts of a "reversible process" and "entropy". For this purpose, an adiabatic irreversible expansion or compression is analysed, by considering that an ideal gas is expanded (compressed), from an initial pressure P[subscript i] to a final pressure P[subscript f], by being placed in…

  11. Pressure-Volume Integral Expressions for Work in Irreversible Processes

    ERIC Educational Resources Information Center

    Gislason, Eric A.; Craig, Norman C.

    2007-01-01

    Different formulations of thermodynamic work "w" as a pressure-volume integral are examined for a piston moving against a gas in an irreversible process. Proper expressions are obtained using the instantaneous pressure of the gas on the piston as the integrand and also using certain external pressures as the integrand. There are two common yet…

  12. Effective Chemical Inactivation of Ebola Virus

    PubMed Central

    Haddock, Elaine; Feldmann, Friederike

    2016-01-01

    Reliable inactivation of specimens before removal from high-level biocontainment is crucial for safe operation. To evaluate efficacy of methods of chemical inactivation, we compared in vitro and in vivo approaches using Ebola virus as a surrogate pathogen. Consequently, we have established parameters and protocols leading to reliable and effective inactivation. PMID:27070504

  13. The Intrinsically Disordered Domain of the Antitoxin Phd Chaperones the Toxin Doc against Irreversible Inactivation and Misfolding*

    PubMed Central

    De Gieter, Steven; Konijnenberg, Albert; Talavera, Ariel; Butterer, Annika; Haesaerts, Sarah; De Greve, Henri; Sobott, Frank; Loris, Remy; Garcia-Pino, Abel

    2014-01-01

    The toxin Doc from the phd/doc toxin-antitoxin module targets the cellular translation machinery and is inhibited by its antitoxin partner Phd. Here we show that Phd also functions as a chaperone, keeping Doc in an active, correctly folded conformation. In the absence of Phd, Doc exists in a relatively expanded state that is prone to dimerization through domain swapping with its active site loop acting as hinge region. The domain-swapped dimer is not capable of arresting protein synthesis in vitro, whereas the Doc monomer is. Upon binding to Phd, Doc becomes more compact and is secured in its monomeric state with a neutralized active site. PMID:25326388

  14. Peptidyl inverse esters of p-methoxybenzoic acid: a novel class of potent inactivator of the serine proteases.

    PubMed Central

    Lynas, J; Walker, B

    1997-01-01

    A series of novel synthetic peptides, containing a C-terminal beta-amino alcohol linked to p-methoxybenzoic acid via an ester linkage, have been prepared and tested as inhibitors against typical members of the serine protease family. For example, the sequences Ac-Val-Pro-NH-CH-(CH2-C6H5)-CH2O-CO-C6H4-OCH3 (I) and Ac-Val-Pro-NH-CH-[CH-(CH3)2]-CH2O-CO-C6H4-OCH3 (II), which fulfil the known primary and secondary specificity requirements of chymotrypsin and elastase respectively, have been found to behave as exceptionally potent irreversible inactivators of their respective target protease. Thus I was found to inactivate chymotrypsin with an overall second-order rate constant (k2/Ki) of approx. 6.6x10(6) M-1. s-1, whereas II is an even more potent inactivator of human neutrophil elastase, exhibiting a second-order rate constant of inactivation of approx. 1.3x10(7) M-1.s-1. These values represent the largest rate constants ever reported for the inactivation of these proteases with synthetic peptide-based inactivators. On prolonged incubation in substrate-containing buffers, samples of the inactivated proteases were found to regain activity slowly. The first-order rate constants for the regeneration of enzymic activity from chymotrypsin and human neutrophil elastase inactivated by I and II respectively were determined to be approx. 5.8x10(-5) s-1 and approx. 4.3x10(-4) s-1. We believe that the most likely mechanism for the inactivation and regeneration of enzymic activity involves the formation and subsequent slow hydrolysis of long-lived acyl enzyme intermediates. PMID:9271079

  15. [The effect of charged local anesthetics on the inactivation of Ca2+-activated Cl-channels of characean algae].

    PubMed

    Kataev, A A; Zherelova, O M; Berestovskiĭ, G N

    1988-01-01

    Effects of local anesthetics (LA) and a number of organic cations on Ca2+-activated Cl-channels in plasmalemma of intracellularly perfused giant algae Nitellopsis obtusa were studied using voltage-clamp technique. It was shown earlier that Ca2+ ions cause irreversible inactivation of Cl-channels with a characteristic time equal to a few minutes, but not only activate Cl-channels. It has been found that amphiphilic cations (AC), including LA+, introduced intracellularly together with Ca2+ produced delayed action on the beginning of the inactivation process (approximately ten minutes) producing no effect on activation during this period. The time of delayed action was linearly dependent on the concentrations ratio alpha = [AC]/[Ca2+]. Procaine is the most effective agent in this respect, the time of its delayed action on the inactivation process being 20 min at alpha = 1. LA in the neural form, hydrophilic AC of tetraethylammonium, as well as LA+ from the outside had no effect on Cl-channels. Cl-channels inactivated "irreversibly" by Ca2+ ions may be restored after addition of AC in Ca2+-containing perfusion medium. PMID:2470412

  16. Kinetic and structural characterization of caspase-3 and caspase-8 inhibition by a novel class of irreversible inhibitors

    SciTech Connect

    Wang, Zhigang; Watt, William; Brooks, Nathan A.; Harris, Melissa S.; Urban, Jan; Boatman, Douglas; McMillan, Michael; Kahn, Michael; Heinrikson, Robert L.; Finzel, Barry C.; Wittwer, Arthur J.; Blinn, James; Kamtekar, Satwik; Tomasselli, Alfredo G.

    2010-09-17

    Because of their central role in programmed cell death, the caspases are attractive targets for developing new therapeutics against cancer and autoimmunity, myocardial infarction and ischemic damage, and neurodegenerative diseases. We chose to target caspase-3, an executioner caspase, and caspase-8, an initiator caspase, based on the vast amount of information linking their functions to diseases. Through a structure-based drug design approach, a number of novel {beta}-strand peptidomimetic compounds were synthesized. Kinetic studies of caspase-3 and caspase-8 inhibition were carried out with these urazole ring-containing irreversible peptidomimetics and a known irreversible caspase inhibitor, Z-VAD-fmk. Using a stopped-flow fluorescence assay, we were able to determine individual kinetic parameters of caspase-3 and caspase-8 inhibition by these inhibitors. Z-VAD-fmk and the peptidomimetic inhibitors inhibit caspase-3 and caspase-8 via a three-step kinetic mechanism. Inhibition of both caspase-3 and caspase-8 by Z-VAD-fmk and of caspase-3 by the peptidomimetic inhibitors proceeds via two rapid equilibrium steps followed by a relatively fast inactivation step. However, caspase-8 inhibition by the peptidomimetics goes through a rapid equilibrium step, a slow-binding reversible step, and an extremely slow inactivation step. The crystal structures of inhibitor complexes of caspases-3 and -8 validate the design of the inhibitors by illustrating in detail how they mimic peptide substrates. One of the caspase-8 structures also shows binding at a secondary, allosteric site, providing a possible route to the development of noncovalent small molecule modulators of caspase activity.

  17. Thiuram Disulfides as Pseudo-irreversible Inhibitors of the Lymphoid Tyrosine Phosphatase

    PubMed Central

    Kulkarni, Rhushikesh A.; Stanford, Stephanie M.; Vellore, Nadeem A.; Krishnamurthy, Divya; Bliss, Matthew R.; Baron, Riccardo; Bottini, Nunzio; Barrios, Amy M.

    2013-01-01

    We have screened a small library of thiuram disulfides for inhibition of lymphoid tyrosine phosphatase (LYP) activity. The parent thiuram disulfide, disulfiram, inhibited LYP activity in vitro and in Jurkat T cells whereas diethyldithiocarbamate failed to inihibit LYP at the concentrations tested. Compound 13, an N-(2-thioxothiazolidin-4-one) analog, was the most potent LYP inhibitor in this series, with an IC50 of 3 μM. Compound 13 was found to inhibit LYP pseudo-irreversibly, as evident by the time-dependence of inhibition with a Ki of 1.1 μM and a kinact of 0.004 s−1. The inhibition of LYP by compound 13 could not be reversed significantly by incubation with glutathione or by prolonged dialysis, but could be partially reversed by incubation with dithiothreitol. Compound 13 also inhibited LYP activity in Jurkat T cells. PMID:23873737

  18. Thiuram disulfides as pseudo-irreversible inhibitors of lymphoid tyrosine phosphatase.

    PubMed

    Kulkarni, Rhushikesh A; Stanford, Stephanie M; Vellore, Nadeem A; Krishnamurthy, Divya; Bliss, Matthew R; Baron, Riccardo; Bottini, Nunzio; Barrios, Amy M

    2013-09-01

    We screened a small library of thiuram disulfides for inhibition of lymphoid tyrosine phosphatase (LYP) activity. The parent thiuram disulfide, disulfiram, inhibited LYP activity in vitro and in Jurkat T cells, whereas diethyldithiocarbamate failed to inhibit LYP at the concentrations tested. Compound 13, an N-(2-thioxothiazolidin-4-one) analogue, was found to be the most potent LYP inhibitor in this series, with an IC50 value of 3 μM. Compound 13 inhibits LYP pseudo-irreversibly, as evidenced by the time-dependence of inhibition, with a K(i) value of 1.1 μM and a k(inact) value of 0.004 s⁻¹. The inhibition of LYP by compound 13 could not be reversed significantly by incubation with glutathione or by prolonged dialysis, but could be partially reversed by incubation with dithiothreitol. Compound 13 also inhibited LYP activity in Jurkat T cells.

  19. The Law of Self-Acting Machines and Irreversible Processes with Reversible Replicas

    NASA Astrophysics Data System (ADS)

    Valev, Pentcho

    2002-11-01

    Clausius and Kelvin saved Carnot theorem and developed the second law by assuming that Carnot machines can work in the absence of an operator and that all the irreversible processes have reversible replicas. The former assumption restored Carnot theorem as an experience of mankind whereas the latter generated "the law of ever increasing entropy". Both assumptions are wrong so it makes sense to return to Carnot theorem (or some equivalent) and test it experimentally. Two testable paradigms - the system performing two types of reversible work and the system in dynamical equilibrium - suggest that perpetuum mobile of the second kind in the presence of an operator is possible. The deviation from the second law prediction, expressed as difference between partial derivatives in a Maxwell relation, measures the degree of structural-functional evolution for the respective system.

  20. Random X-chromosome inactivation: skewing lessons for mice and men.

    PubMed

    Clerc, Philippe; Avner, Philip

    2006-06-01

    The mammalian X-chromosome exists in two flavors, active and inactive, in each cell of the adult female. This phenomenon originates from the process of random choice occurring early in development in a small number of progenitor cells in which the decision is made to inactivate either one or the other X chromosome on a cell-autonomous basis. Once made, this initial decision is irreversible, although exceptions exist in specific chromosomal territories and cell lineages. Recent findings implicate various factors, including non-coding RNAs and chromatin modification complexes, as effectors in the initiation and maintenance of X-chromosome inactivation. The functional redundancy of such factors almost certainly plays an important role in the stability of the inactive X. Studying skewing or bias opens an important opportunity for understanding facets of the random choice process.

  1. Caspase 3-Mediated Inactivation of Rac GTPases Promotes Drug-Induced Apoptosis in Human Lymphoma Cells

    PubMed Central

    Zhang, Baolin; Zhang, Yaqin; Shacter, Emily

    2003-01-01

    The Rac members of the Rho family GTPases control signaling pathways that regulate diverse cellular activities, including cytoskeletal organization, gene transcription, and cell transformation. Rac is implicated in apoptosis, but little is known about the mechanism by which it responds to apoptotic stimuli. Here we demonstrate that endogenous Rac GTPases are caspase 3 substrates that are cleaved in human lymphoma cells during drug-induced apoptosis. Cleavage of Rac1 occurs at two unconventional caspase 3 sites, VVGD11/G and VMVD47/G, and results in inactivation of the GTPase and effector functions of the protein (binding to the p21-activated protein kinase PAK1). Expression of caspase 3-resistant Rac1 mutants in the cells suppresses drug-induced apoptosis. Thus, proteolytic inactivation of Rac GTPases represents a novel, irreversible mechanism of Rac downregulation that allows maximal cell death following drug treatment. PMID:12897143

  2. BNNT-mediated irreversible electroporatio: its potential on cancer cells

    SciTech Connect

    Vittoria Raffa, Cristina Riggio, Michael W. Smith, Kevin C. Jordan, Wei Cao, Alfred Cuschieri

    2012-10-01

    Tissue ablation, i.e., the destruction of undesirable tissues, has become an important minimally invasive technique alternative to resection surgery for the treatment of tumours. Several methods for tissue ablation are based on thermal techniques using cold, e.g. cryosurgery [1] or heat, e.g. radiofrequency [2] or high-intensity focused ultrasound [3] or nanoparticle-mediated irradiation [4]. Alternatively, irreversible electroporation (IRE) has been proposed as non thermal technique for minimally invasive tissue ablation based on the use of electrical pulses. When the electric field is applied to a cell, a change in transmembrane potential is induced, which can cause biochemical and physiological changes of the cell. When the threshold value of the transmembrane potential is exceeded, the cell membrane becomes permeable, thus allowing entrance of molecules that otherwise cannot cross the membrane [5]. A further increase in the electric field intensity may cause irreversible membrane permeabilization and cell death. These pulses create irreversible defects (pores) in the cell membrane lipid bilayer, causing cell death through loss of cell homeostasis [6]. This is desirable in tumour ablation in order to produce large cell death, without the use of cytostatic drugs. A study of Davalos, Mir and Rubinsky showed that IRE can ablate substantial volumes of tissue without inducing a thermal effect and therefore serve as an independent and new tissue ablation modality; this opened the way to the use of IRE in surgery [7]. Their finding was subsequently confirmed in studies on cells [8], small animal models [9] and in large animal models in the liver [10] and the heart [11]. The most important finding in these papers is that irreversible electroporation produces precisely delineated ablation zones with cell scale resolution between ablated and non-ablated areas, without zones in which the extent of damage changes gradually as during thermal ablation. Furthermore, it is

  3. Inactivation of allergens and toxins.

    PubMed

    Morandini, Piero

    2010-11-30

    Plants are replete with thousands of proteins and small molecules, many of which are species-specific, poisonous or dangerous. Over time humans have learned to avoid dangerous plants or inactivate many toxic components in food plants, but there is still room for ameliorating food crops (and plants in general) in terms of their allergens and toxins content, especially in their edible parts. Inactivation at the genetic rather than physical or chemical level has many advantages and classical genetic approaches have resulted in significant reduction of toxin content. The capacity, offered by genetic engineering, of turning off (inactivating) specific genes has opened up the possibility of altering the plant content in a far more precise manner than previously available. Different levels of intervention (genes coding for toxins/allergens or for enzymes, transporters or regulators involved in their metabolism) are possible and there are several tools for inactivating genes, both direct (using chemical and physical mutagens, insertion of transposons and other genetic elements) and indirect (antisense RNA, RNA interference, microRNA, eventually leading to gene silencing). Each level/strategy has specific advantages and disadvantages (speed, costs, selectivity, stability, reversibility, frequency of desired genotype and regulatory regime). Paradigmatic examples from classical and transgenic approaches are discussed to emphasize the need to revise the present regulatory process. Reducing the content of natural toxins is a trade-off process: the lesser the content of natural toxins, the higher the susceptibility of a plant to pests and therefore the stronger the need to protect plants. As a consequence, more specific pesticides like Bt are needed to substitute for general pesticides.

  4. Selective and irreversible inhibitors of mosquito acetylcholinesterases for controlling malaria and other mosquito-borne diseases.

    PubMed

    Pang, Yuan-Ping; Ekström, Fredrik; Polsinelli, Gregory A; Gao, Yang; Rana, Sandeep; Hua, Duy H; Andersson, Björn; Andersson, Per Ola; Peng, Lei; Singh, Sanjay K; Mishra, Rajesh K; Zhu, Kun Yan; Fallon, Ann M; Ragsdale, David W; Brimijoin, Stephen

    2009-01-01

    New insecticides are urgently needed because resistance to current insecticides allows resurgence of disease-transmitting mosquitoes while concerns for human toxicity from current compounds are growing. We previously reported the finding of a free cysteine (Cys) residue at the entrance of the active site of acetylcholinesterase (AChE) in some insects but not in mammals, birds, and fish. These insects have two AChE genes (AP and AO), and only AP-AChE carries the Cys residue. Most of these insects are disease vectors such as the African malaria mosquito (Anopheles gambiae sensu stricto) or crop pests such as aphids. Recently we reported a Cys-targeting small molecule that irreversibly inhibited all AChE activity extracted from aphids while an identical exposure caused no effect on the human AChE. Full inhibition of AChE in aphids indicates that AP-AChE contributes most of the enzymatic activity and suggests that the Cys residue might serve as a target for developing better aphicides. It is therefore worth investigating whether the Cys-targeting strategy is applicable to mosquitocides. Herein, we report that, under conditions that spare the human AChE, a methanethiosulfonate-containing molecule at 6 microM irreversibly inhibited 95% of the AChE activity extracted from An. gambiae s. str. and >80% of the activity from the yellow fever mosquito (Aedes aegypti L.) or the northern house mosquito (Culex pipiens L.) that is a vector of St. Louis encephalitis. This type of inhibition is fast ( approximately 30 min) and due to conjugation of the inhibitor to the active-site Cys of mosquito AP-AChE, according to our observed reactivation of the methanethiosulfonate-inhibited AChE by 2-mercaptoethanol. We also note that our sulfhydryl agents partially and irreversibly inhibited the human AChE after prolonged exposure (>4 hr). This slow inhibition is due to partial enzyme denaturation by the inhibitor and/or micelles of the inhibitor, according to our studies using atomic force

  5. Hydrazine vapor inactivates Bacillus spores

    NASA Astrophysics Data System (ADS)

    Schubert, Wayne W.; Engler, Diane L.; Beaudet, Robert A.

    2016-05-01

    NASA policy restricts the total number of bacterial spores that can remain on a spacecraft traveling to any planetary body which might harbor life or have evidence of past life. Hydrazine, N2H4, is commonly used as a propellant on spacecraft. Hydrazine as a liquid is known to inactivate bacterial spores. We have now verified that hydrazine vapor also inactivates bacterial spores. After Bacillus atrophaeus ATCC 9372 spores deposited on stainless steel coupons were exposed to saturated hydrazine vapor in closed containers, the spores were recovered from the coupons, serially diluted, pour plated and the surviving bacterial colonies were counted. The exposure times required to reduce the spore population by a factor of ten, known as the D-value, were 4.70 ± 0.50 h at 25 °C and 2.85 ± 0.13 h at 35 °C. These inactivation rates are short enough to ensure that the bioburden of the surfaces and volumes would be negligible after prolonged exposure to hydrazine vapor. Thus, all the propellant tubing and internal tank surfaces exposed to hydrazine vapor do not contribute to the total spore count.

  6. Inactivation of Cephalosporins by Bacteroides

    PubMed Central

    Tally, Francis P.; O'Keefe, J. Paul; Sullivan, N. M.; Gorbach, Sherwood L.

    1979-01-01

    We investigated the relationship between β-lactamases of Bacteroides fragilis organisms and their resistance to cephalosporins. Timed killing curves were used to study the in vitro activity of three cephalosporins, cephalothin, cefazolin, and cefamandole, and a semisynthetic cephamycin, cefoxitin. Measurements of residual antibiotic concentrations in culture supernatants were made, and they were compared with the β-lactamase activity of the microorganism. A cephalosporin-susceptible strain was rapidly killed by cephalothin, cefazolin, cefamandole, and cefoxitin. Four cephalosporin-resistant strains were not killed by cephalothin, cefazolin, or cefamandole but were killed by cefoxitin. An inoculum effect was noted with cefazolin and not with cefoxitin. The resistant strains of Bacteroides inactivated the three cephalosporins, but there was no inactivation of cefoxitin. A constitutive β-lactamase was detected in all the isolates of the B. fragilis group that were resistant to the cephalosporins. There was no distinction of the species based on isoelectric focusing of the enzyme. These data suggest that inactivation by β-lactamase may be the mechanism for resistance of B. fragilis to the cephalosporins and would explain the enhanced in vitro activity of cefoxitin. PMID:525995

  7. Suicidal inactivation of methemoglobin by generation of thiyl radical: insight into NAC mediated protection in RBC.

    PubMed

    Balaji, S N; Trivedi, V

    2013-07-01

    N-acetyl-L-cysteine (NAC) improves antioxidant potentials of RBCs to provide protection against oxidative stress induced hemolysis. The antioxidant mechanism of NAC to reduce oxidative stress in RBC, studied through inactivation of pro-oxidant MetHb. NAC causes irreversible inactivation of the MetHb in an H2O2 dependent manner, and the inactivation follows the pseudo- first- order kinetics. The kinetic constants are ki = 8.5μM, kinact = 0.706 min(-1) and t1/2 = 0.9 min. Spectroscopic studies indicate that MetHb accepts NAC as a substrate and oxidizes through a single electron transfer mechanism to the NACox. The single e- oxidation product of NAC has been identified as the 5, 5'- dimethyl-1- pyrroline N- oxide (DMPO) adduct of the sulfur centered radical (a(N) = 15.2 G and a(H)=16.78 G). Binding studies indicate that NACox interacts at the heme moiety and NAC oxidation through MetHb is essential for NAC binding. Heme-NAC adduct dissociated from MetHb and identified (m/z 1011.19) as 2:1 ratio of NAC/heme in the adduct. TEMPO and PBN treatment reduces NAC binding to MetHb and protects against inactivation confirms the role of thiyl radical in the inactivation process. Furthermore, scavenging thiyl radicals by TEMPO abolish the protective effect of NAC in hemolysis. Current work highlights antioxidant mechanism of NAC through NAC thiyl radical generation, and MetHb inactivation to exhibit protection in RBC against oxidative stress induced hemolysis.

  8. Inactivation of tyrosine hydroxylase activity by ascorbate in vitro and in rat PC12 cells.

    PubMed

    Wilgus, H; Roskoski, R

    1988-10-01

    Tyrosine hydroxylase activity is reversibly modulated by the actions of a number of protein kinases and phosphoprotein phosphatases. A previous report from this laboratory showed that low-molecular-weight substances present in striatal extracts lead to an irreversible loss of tyrosine hydroxylase activity under cyclic AMP-dependent phosphorylation conditions. We report here that ascorbate is one agent that inactivates striatal tyrosine hydroxylase activity with an EC50 of 5.9 microM under phosphorylating conditions. Much higher concentrations (100 mM) fail to inactivate the enzyme under nonphosphorylating conditions. Isoascorbate (EC50, 11 microM) and dehydroascorbate (EC50, 970 microM) also inactivated tyrosine hydroxylase under phosphorylating but not under nonphosphorylating conditions. In contrast, ascorbate sulfate was inactive under phosphorylating conditions at concentrations up to 100 mM. Since the reduced compounds generate several reactive species in the presence of oxygen, the possible protecting effects of catalase, peroxidase, and superoxide dismutase were examined. None of these three enzymes, however, afforded any protection against inactivation. We also examined the effects of ascorbate and its congeners on the activity of tyrosine hydroxylase purified to near homogeneity from a rat pheochromocytoma. This purified enzyme was also inactivated by the same agents that inactivated the impure corpus striatal enzyme. Under conditions in which ascorbate almost completely abolished enzyme activity, we found no indication for significant proteolysis of the purified enzyme as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. We also found that pretreatment of PC12 cells in culture for 4 h with 1 mM ascorbate, dehydroascorbate, or isoascorbate (but not ascorbate sulfate) also decreased tyrosine hydroxylase activity 25-50%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:2901463

  9. Aftereffect in rocks caused by preexisting irreversible deformations

    SciTech Connect

    Stavrogin, A.N.; Shirkes, O.A.

    1987-05-01

    In this paper, rock specimens cut as cores of a diameter of 30 mm, 80 mm in length, were subjected to irreversible deformation in a high hydrostatic pressure chamber according to Karman's procedure. The types of rocks investigated were white Koelga marble, non-burst-hazardous (NBH) sandstone from Donets Basin, limestone from Estonslanets deposit and brown coal from Shurab coal deposit. Marble specimens were subjected to the most extensive studies. The aftereffect curves are shown for each type of rock studied. Aftereffect deformations of rocks are basically creep flows occurring under the effect of residual stresses introduced into the rock material on the course of its irreversible deformation by a high hydrostatic pressure, according to the authors. The physical nature of the residual stresses in the rocks and the mechanism of their creation are examined at the level of structural elements (grains or crystals).

  10. Flux pinning characteristics and irreversibility line in high temperature superconductors

    NASA Technical Reports Server (NTRS)

    Matsushita, T.; Ihara, N.; Kiuchi, M.

    1995-01-01

    The flux pinning properties in high temperature superconductors are strongly influenced by thermally activated flux motion. The scaling relation of the pinning force density and the irreversibility line in various high temperature superconductors are numerically analyzed in terms of the flux creep model. The effect of two factors, i.e., the flux pinning strength and the dimensionality of the material, on these properties are investigated. It is speculated that the irreversibility line in Bi-2212 superconductors is one order of magnitude smaller than that in Y-123, even if the flux pinning strength in Bi-2212 is improved up to the level of Y-123. It is concluded that these two factors are equally important in determination of the flux pinning characteristics at high temperatures.

  11. Flux pinning characteristics and irreversibility line in high temperature superconductors

    SciTech Connect

    Matsushita, T.; Ihara, N.; Kiuchi, M.

    1995-04-01

    The flux pinning properties in high temperature superconductors are strongly influenced by thermally activated flux motion. The scaling relation of the pinning force density and the irreversibility line in various high temperature superconductors are numerically analyzed in terms of the flux creep model. The effect of two factors, i.e., the flux pinning strength and the dimensionality of the material, on these properties are investigated. It is speculated that the irreversibility line in Bi-2212 superconductors is one order of magnitude smaller than that in Y-123, even if the flux pinning strength in Bi-2212 is improved up to the level of Y-123. It is concluded that these two factors are equally important in determination of the flux pinning characteristics at high temperatures.

  12. Prostaglandin E2 to diagnose between reversible and irreversible pulpitis.

    PubMed

    Petrini, M; Ferrante, M; Ciavarelli, L; Brunetti, L; Vacca, M; Spoto, G

    2012-01-01

    The aim of this work is to verify a correlation between the grade of inflammation and the concentration of PGE2 in human dental pulp. A total of 25 human dental pulps were examined by histological analysis and radioimmunologic dosage of PGE2. The pulps used in this experiment were from healthy and symptomatic teeth; the first ones were collected from teeth destined to be extracted for orthodontic reasons. An increase was observed of PGE2 in reversible pulpitis compared with healthy pulps and with the irreversible pulpitis and the clear decrease of these when NSAIDs are taken. This study demonstrates that PGE2 level is correlated to histological analysis thus allowing to distinguish symptomatic teeth in reversible and irreversible pulpitis. PMID:22507328

  13. Irreversible adsorption of phenolic compounds by activated carbons

    SciTech Connect

    Grant, T.M.; King, C.J.

    1988-12-01

    Studies were undertaken to determine the reasons why phenolic sorbates can be difficult to remove and recover from activated carbons. The chemical properties of the sorbate and the adsorbent surface, and the influences of changes in the adsorption and desorption conditions were investigated. Comparison of isotherms established after different contact times or at different temperatures indicated that phenolic compounds react on carbon surfaces. The reaction rate is a strong function of temperature. Regeneration of carbons by leaching with acetone recovered at least as much phenol as did regeneration with other solvents or with displacers. The physiochemical properties of adsorbents influences irreversible uptakes. Sorbates differed markedly in their tendencies to undergo irreversible adsorption. 64 refs., 47 figs., 32 tabs.

  14. A Comparison of Reversible Versus Irreversible Protein Glutathionylation

    PubMed Central

    Townsend, Danyelle M.; Lushchak, Volodymyr I.; Cooper, Arthur J.L.

    2015-01-01

    Glutathionylation is generally a reversible posttranslational modification that occurs to cysteine residues that have been exposed to reactive oxygen species (P-SSG). This cyclical process can regulate various clusters of proteins, including those involved in critical cellular signaling functions. However, certain conditions can favor the formation of dehydroamino acids, such as 2,3-didehydroalanine (2,3-dehydroalanine, DHA) and 2,3-didehydrobutyrine (2,3-dehydrobutyrine), which can act as Michael acceptors. In turn, these can form Michael adducts with glutathione (GSH), resulting in the formation of a stable thioether conjugate, an irreversible process referred to as nonreducible glutathionylation. This is predicted to be prevalent in nature, particularly in more slowly turning over proteins. Such nonreducible glutathionylation can be distinguished from the more facile cycling signaling processes and is predicted to be of gerontological, toxicological, pharmacological, and oncological relevance. Here, we compare reversible and irreversible glutathionylation. PMID:24974182

  15. Thermoeconomic analysis of an irreversible Stirling heat pump cycle

    NASA Astrophysics Data System (ADS)

    Lucia, U.; Gervino, G.

    2006-03-01

    In this paper an analysis of the Stirling cycle in thermoeconomic terms is developed using the entropy generation. In the thermoeconomic optimization of an irreversible Stirling heat pump cycle the F function has been introduced to evaluate the optimum for the higher and lower sources temperature ratio in the cycle: this ratio represents the value which optimizes the cycle itself. The variation of the function F is proportional to the variation of the entropy generation, the maxima and minima of F has been evaluated in a previous paper without giving the physical foundation of the method. We investigate the groundwork of this approach: to study the upper and lower limits of F function allows to determine the cycle stability and the optimization conditions. The optimization consists in the best COP at the least cost. The principle of maximum variation for the entropy generation becomes the analytic foundation of the optimization method in the thermoeconomic analysis for an irreversible Stirling heat pump cycle.

  16. Properties and kinetic analysis of UDP-glucose dehydrogenase from group A streptococci. Irreversible inhibition by UDP-chloroacetol.

    PubMed

    Campbell, R E; Sala, R F; van de Rijn, I; Tanner, M E

    1997-02-01

    UDP-glucuronic acid is used by many pathogenic bacteria in the construction of an antiphagocytic capsule that is required for virulence. The enzyme UDP-glucose dehydrogenase catalyzes the NAD+-dependent 2-fold oxidation of UDP-glucose and provides a source of the acid. In the present study the recombinant dehydrogenase from group A streptococci has been purified and found to be active as a monomer. The enzyme contains no chromophoric cofactors, and its activity is unaffected by the presence of EDTA or carbonyl-trapping reagents. Initial velocity and product inhibition kinetic patterns are consistent with a bi-uni-uni-bi ping-pong mechanism in which UDP-glucose is bound first and UDP-glucuronate is released last. UDP-xylose was found to be a competitive inhibitor (Ki, 2.7 microM) of the enzyme. The enzyme is irreversibly inactivated by uridine 5'-diphosphate-chloroacetol due to the alkylation of an active site cysteine thiol. The apparent second order rate constant for the inhibition (ki/Ki) was found to be 2 x 10(3) mM-1 min-1. Incubation with the truncated compound, chloroacetol phosphate, resulted in no detectable inactivation when tested under comparable conditions. This supports the notion that uridine 5'-diphosphate-chloroacetol is bound in the place of UDP-glucose and is not simply acting as a nonspecific alkylating agent.

  17. Variability of Irreversible Poleward Transport in the Lower Stratosphere

    NASA Technical Reports Server (NTRS)

    Olsen, Mark; Douglass, Anne; Newman, Paul; Nash, Eric; Witte, Jacquelyn; Ziemke, Jerry

    2011-01-01

    The ascent and descent of the Brewer-Dobson circulation plays a large role in determining the distributions of many constituents in the extratropical lower stratosphere. However, relatively fast, quasi-horizontal transport out of the tropics and polar regions also significantly contribute to determining these distributions. The tropical tape recorder signal assures that there must be outflow from the tropics into the extratropical lower stratosphere. The phase of the quasi-biennial oscillation (QBO) and state of the polar vortex are known to modulate the transport from the tropical and polar regions, respectively. In this study we examine multiple years of ozone distributions in the extratropical lower stratosphere observed by the Aura Microwave Limb Sounder (MLS) and the Aura High Resolution Dynamic Limb Sounder (HIRDLS). The distributions are compared with analyses of irreversible, meridional isentropic transport. We show that there is considerable year-to-year seasonal variability in the amount of irreversible transport from the tropics, which is related to both the phase of the QBO and the state of the polar vortex. The reversibility of the transport is consistent with the number of observed breaking waves. The variability of the atmospheric index of refraction in the lower stratosphere is shown to be significantly correlated with the wave breaking and amount of irreversible transport. Finally, we will show that the seasonal extratropical stratosphere to troposphere transport of ozone can be substantially modulated by the amount of irreversible meridional transport in the lower stratosphere and we investigate how observable these differences are in data of tropospheric ozone.

  18. Irreversibility and chaos: role of lubrication interactions in sheared suspensions.

    PubMed

    Metzger, Bloen; Pham, Phong; Butler, Jason E

    2013-05-01

    We investigate non-Brownian particles suspended in a periodic shear-flow using simulations. Following Metzger and Butler [Phys. Rev. E 82, 051406 (2010)], we show that the chaotic dynamics arising from lubrication interactions are too weak to generate an observable particle dispersion. The irreversibility observed in periodic flow is dominated by contact interactions. Nonetheless, we show that lubrication interactions must be included in the calculation to obtain results that agree with experiments.

  19. Thermodynamical estimation of the limit potentialities of irreversible binary distillation

    NASA Astrophysics Data System (ADS)

    Tsirlin, A. M.; Grigorevsky, I. N.

    2010-10-01

    The limit capacity of binary distillation is considered for conventional heat supply to the column bottom and heat removal from the refluxer and for heat supply and removal distributed over the column height. The form of operating line, appropriating to the minimum of irreversible loss for adjusted productivity and limited transfer coefficients, was found. The expression for possible capacity limit of the distillation column was obtained, depending on flow composition and heat and mass transfer coefficients.

  20. Irreversibility and complex network behavior of stream flow fluctuations

    NASA Astrophysics Data System (ADS)

    Serinaldi, Francesco; Kilsby, Chris G.

    2016-05-01

    Exploiting the duality between time series and networks, directed horizontal visibility graphs (DHVGs) are used to perform an unprecedented analysis of the dynamics of stream flow fluctuations with focus on time irreversibility and long range dependence. The analysis relies on a large quality-controlled data set consisting of 699 daily time series recorded in the continental United States (CONUS) that are not affected by human activity and primarily reflects meteorological conditions. DHVGs allow a clear visualization and quantification of time irreversibility of flow dynamics, which can be interpreted as a signature of nonlinearity, and long range dependence resulting from the interaction of atmospheric, surface and underground processes acting at multiple spatio-temporal scales. Irreversibility is explored by mapping the time series into ingoing, outgoing, and undirected graphs and comparing the corresponding degree distributions. Using surrogate data preserving up to the second order linear temporal dependence properties of the observed series, DHVGs highlight the additional complexity introduced by nonlinearity into flow fluctuation dynamics. We show that the degree distributions do not decay exponentially as expected, but tend to follow a subexponential behavior, even though sampling uncertainty does not allow a clear distinction between apparent or true power law decay. These results confirm that the complexity of stream flow dynamics goes beyond a linear representation involving for instance the combination of linear processes with short and long range dependence, and requires modeling strategies accounting for temporal asymmetry and nonlinearity.

  1. Anesthetic Efficacy in Irreversible Pulpitis: A Randomized Clinical Trial.

    PubMed

    Allegretti, Carlos E; Sampaio, Roberta M; Horliana, Anna C R T; Armonia, Paschoal L; Rocha, Rodney G; Tortamano, Isabel Peixoto

    2016-01-01

    Inferior alveolar nerve block has a high failure rate in the treatment of mandibular posterior teeth with irreversible pulpitis. The aim of this study was to compare the anesthetic efficacy of 4% articaine, 2% lidocaine and 2% mepivacaine, all in combination with 1:100,000 epinephrine, in patients with irreversible pulpitis of permanent mandibular molars during a pulpectomy procedure. Sixty-six volunteers from the Emergency Center of the School of Dentistry, University of São Paulo, randomly received 3.6 mL of local anesthetic as a conventional inferior alveolar nerve block (IANB). The subjective signal of lip numbness, pulpal anesthesia and absence of pain during the pulpectomy procedure were evaluated respectively, by questioning the patient, stimulation using an electric pulp tester and a verbal analogue scale. All patients reported the subjective signal of lip numbness. Regarding pulpal anesthesia success as measured with the pulp tester, the success rate was respectively 68.2% for mepivacaine, 63.6% for articaine and 63.6% for lidocaine. Regarding patients who reported no pain or mild pain during the pulpectomy, the success rate was, respectively 72.7% for mepivacaine, 63.6% for articaine and 54.5% for lidocaine. These differences were not statistically significant. Neither of the solutions resulted in 100% anesthetic success in patients with irreversible pulpitis of mandibular molars.

  2. Irreversible steps in the ferritin synthesis induction pathway.

    PubMed

    Goessling, L S; Mascotti, D P; Bhattacharyya-Pakrasi, M; Gang, H; Thach, R E

    1994-02-11

    The ability of cells to re-repress ferritin synthesis after removal of an inducing agent (iron or heme) was investigated. Re-repression was found to be a slow process, requiring approximately 4 (after iron removal) to 10 h (after heme removal) for completion. Desferrioxamine mesylate (Desferal) had only a slight effect on the rate of re-repression, whereas cycloheximide was strongly inhibitory, indicating that new protein synthesis is required for re-repression. Re-repression occurred at a slow but significant rate in the presence of both Desferal and cycloheximide. These results indicate that, in the absence of an iron chelator, the induction of ferritin synthesis is essentially irreversible. The kinetics of the previously reported covalent modification of IRE-binding protein (IRE-BP) were then examined, to see whether this phenomenon might account (at least in part) for the irreversibility of induction. It was found that the heme- or iron-dependent disappearance of 98-kDa IRE-BP occurred rapidly (within 1 h), and was equally rapidly reversed upon removal of heme after a 1-h exposure. By contrast, after a 4-h exposure to heme, little 98-kDa IRE-BP could be regenerated after heme removal. These results suggest that the slow, irreversible covalent modification of IRE-BP correlates closely over time with the induction of ferritin synthesis. The covalent modification of IRE-BP depends on cell growth rate, and is most readily detected in rapidly growing cells.

  3. Developing irreversible inhibitors of the protein kinase cysteinome

    PubMed Central

    Liu, Qingsong; Sabnis, Yogesh; Zhao, Zheng; Zhang, Tinghu; Buhrlage, Sara J.; Jones, Lyn H.; Gray, Nathanael S.

    2013-01-01

    Protein kinases are a large family of approximately 530 highly conserved enzymes that transfer a γ-phosphate group from ATP to a variety of amino acid residues such as tyrosine, serine and threonine which serves as a ubiquitous mechanism for cellular signal transduction. The clinical success of a number of kinase-directed drugs and the frequent observation of disease causing mutations in protein kinases suggest that a large number of kinases may represent therapeutically relevant targets. To-date the majority of clinical and preclinical kinase inhibitors are ATP-competitive, non-covalent inhibitors that achieve selectivity through recognition of unique features of particular protein kinases. Recently there has been renewed interest in the development of irreversible inhibitors that form covalent bonds with cysteine or other nucleophilic residues in the ATP-binding pocket. Irreversible kinase inhibitors have a number of potential advantages including prolonged pharmacodynamics, suitability for rational design, high potency and ability to validate pharmacological specificity through mutation of the reactive cysteine residue. Here we review recent efforts to develop cysteine-targeted irreversible protein kinase inhibitors and discuss their modes of recognizing the ATP-binding pocket and their biological activity profiles. In addition, we provided an informatics assessment of the potential ‘kinase-cysteinome’ and discuss strategies for the efficient development of new covalent inhibitors. PMID:23438744

  4. Irreversible Electroporation for Focal Ablation at the Porta Hepatis

    SciTech Connect

    Kasivisvanathan, Veeru; Thapar, Ankur Oskrochi, Youssof; Picard, John; Leen, Edward L. S.

    2012-12-15

    Patients with chemotherapy-refractory liver metastases who are not candidates for surgery may be treated with focal ablation techniques with established survival benefits. Irreversible electroporation is the newest of these and has the putative advantages of a nonthermal action, preventing damage to adjacent biliary structures and bowel. This report describes the use of irreversible electroporation in a 61-year-old man with a solitary chemoresistant liver metastasis unsuitable for radiofrequency ablation as a result of its proximity to the porta hepatis. At 3 months, tumor size was decreased on computed tomography from 28 Multiplication-Sign 19 to 20 Multiplication-Sign 17 mm, representing stable disease according to the response evaluation criteria in solid tumors. This corresponded to a decrease in tumor volume size from 5.25 to 3.16 cm{sup 3}. There were no early or late complications. Chemoresistant liver metastases in the proximity of the porta hepatis that are considered to be too high a risk for conventional surgery or thermal ablation may be considered for treatment by the novel ablation technique of irreversible electroporation.

  5. Noncomplementing Diploids from Bacillus Subtilis Protoplast Fusion: Relationship between Maintenance of Chromosomal Inactivation and Segregation Capacity

    PubMed Central

    Grandjean, V.; Hauck, Y.; Le-Derout, J.; Hirschbein, L.

    1996-01-01

    Fusions of Bacillus subtilis protoplasts from two genetically marked strains produce noncomplementing heterodiploid bacteria. These noncomplementing diploids (Ncds) carry both parental chromosomes, but only one is expressed. Fusion products of strains polymorphic for NotI restriction sites provide new physical evidence to support the conclusion that Ncds are not an artifact of cross feeding or cell adhesion. We show that reversible chromosomal inactivation can only account for the biparental trait of unstable Ncds. Two types of cells were recovered from the late progeny of unstable Ncds: Ncds with irreversible chromosome silencing (stable Ncds) and secondary recombinants that displayed a genomic mosaic NotI profile. Segregants from an unstable Ncd population gave rise to two viable haploid cell types. By contrast, stable Ncds segregated into a population of viable and inviable haploid cells. We propose that the latter are derived from irreversible chromosome silencing. Our results indicate that clonal populations of stable Ncds are heterogenous and suggest that segregation and inactivation are independent parameters. PMID:8913734

  6. Urease from Helicobacter pylori is inactivated by sulforaphane and other isothiocyanates

    PubMed Central

    Fahey, Jed W.; Stephenson, Katherine K.; Wade, Kristina L.; Talalay, Paul

    2013-01-01

    Infections by Helicobacter pylori are very common, causing gastroduodenal inflammation including peptic ulcers, and increasing the risk of gastric neoplasia. The isothiocyanate (ITC) sulforaphane [SF; 1-isothiocyanato-4-(methylsulfinyl)butane] derived from edible crucifers such as broccoli is potently bactericidal against Helicobacter, including antibiotic-resistant strains, suggesting a possible dietary therapy. Gastric H. pylori infections express high urease activity which generates ammonia, neutralizes gastric acidity, and promotes inflammation. The finding that SF inhibits (inactivates) urease (jack bean and Helicobacter) raised the issue of whether these properties might be functionally related. The rates of inactivation of urease activity depend on enzyme and SF concentrations and show first order kinetics. Treatment with SF results in time-dependent increases in the ultraviolet absorption of partially purified Helicobacter urease in the 280–340 nm region. This provides direct spectroscopic evidence for the formation of dithiocarbamates between the ITC group of SF and cysteine thiols of urease. The potencies of inactivation of Helicobacter urease by isothiocyanates structurally related to SF were surprisingly variable. Natural isothiocyanates closely related to SF, previously shown to be bactericidal (berteroin, hirsutin, phenethyl isothiocyanate, alyssin, and erucin), did not inactivate urease activity. Furthermore, SF is bactericidal against both urease positive and negative H. pylori strains. In contrast, some isothiocyanates such as benzoyl-ITC, are very potent urease inactivators, but are not bactericidal. The bactericidal effects of SF and other ITC against Helicobacter are therefore not obligatorily linked to urease inactivation, but may reduce the inflammatory component of Helicobacter infections. PMID:23583386

  7. Mechanism of inactivation of alanine racemase by beta, beta, beta-trifluoroalanine

    SciTech Connect

    Faraci, W.S.; Walsh, C.T.

    1989-01-24

    The alanine racemases are a group of PLP-dependent bacterial enzymes that catalyze the racemization of alanine, providing D-alanine for cell wall synthesis. Inactivation of the alanine racemases from the Gram-negative organism Salmonella typhimurium and Gram-positive organism Bacillus stearothermophilus with beta, beta, beta-trifluoroalanine has been studied. The inactivation occurs with the same rate constant as that for formation of a broad 460-490-nm chromophore. Loss of two fluoride ions per mole of inactivated enzyme and retention of (1-/sup 14/C)trifluoroalanine label accompany inhibition, suggesting a monofluoro enzyme adduct. Partial denaturation (1 M guanidine) leads to rapid return of the initial 420-nm chromophore, followed by a slower (t1/2 approximately 30 min-1 h) loss of the fluoride ion and /sup 14/CO/sub 2/ release. At this point, reduction by NaB/sub 3/H/sub 4/ and tryptic digestion yield a single radiolabeled peptide. Purification and sequencing of the peptide reveals that lysine-38 is covalently attached to the PLP cofactor. A mechanism for enzyme inactivation by trifluoroalanine is proposed and contrasted with earlier results on monohaloalanines, in which nucleophilic attack of released aminoacrylate on the PLP aldimine leads to enzyme inactivation. For trifluoroalanine inactivation, nucleophilic attack of lysine-38 on the electrophilic beta-difluoro-alpha, beta-unsaturated imine provides an alternative mode of inhibition for these enzymes.

  8. Urease from Helicobacter pylori is inactivated by sulforaphane and other isothiocyanates.

    PubMed

    Fahey, Jed W; Stephenson, Katherine K; Wade, Kristina L; Talalay, Paul

    2013-05-24

    Infections by Helicobacter pylori are very common, causing gastroduodenal inflammation including peptic ulcers, and increasing the risk of gastric neoplasia. The isothiocyanate (ITC) sulforaphane [SF; 1-isothiocyanato-4-(methylsulfinyl)butane] derived from edible crucifers such as broccoli is potently bactericidal against Helicobacter, including antibiotic-resistant strains, suggesting a possible dietary therapy. Gastric H. pylori infections express high urease activity which generates ammonia, neutralizes gastric acidity, and promotes inflammation. The finding that SF inhibits (inactivates) urease (jack bean and Helicobacter) raised the issue of whether these properties might be functionally related. The rates of inactivation of urease activity depend on enzyme and SF concentrations and show first order kinetics. Treatment with SF results in time-dependent increases in the ultraviolet absorption of partially purified Helicobacter urease in the 260-320 nm region. This provides direct spectroscopic evidence for the formation of dithiocarbamates between the ITC group of SF and cysteine thiols of urease. The potencies of inactivation of Helicobacter urease by isothiocyanates structurally related to SF were surprisingly variable. Natural isothiocyanates closely related to SF, previously shown to be bactericidal (berteroin, hirsutin, phenethyl isothiocyanate, alyssin, and erucin), did not inactivate urease activity. Furthermore, SF is bactericidal against both urease positive and negative H. pylori strains. In contrast, some isothiocyanates such as benzoyl-ITC, are very potent urease inactivators, but are not bactericidal. The bactericidal effects of SF and other ITC against Helicobacter are therefore not obligatorily linked to urease inactivation, but may reduce the inflammatory component of Helicobacter infections. PMID:23583386

  9. Meprin Metalloproteases Inactivate Interleukin 6*

    PubMed Central

    Keiffer, Timothy R.; Bond, Judith S.

    2014-01-01

    Meprins have been implicated in the pathogenesis of several inflammatory diseases, including inflammatory bowel disease, in which the cytokine IL-6 is a prominent effector molecule. Because IL-6 levels are elevated markedly in meprin α and α/β knockout mice in an experimental model of inflammatory bowel disease, the interaction between meprins and IL-6 was studied. The results demonstrate that rodent and human meprin A and B cleave IL-6 to a smaller product and, subsequently, are capable of extensive degradation of the cytokine. Analysis of the limited degradation product formed by meprin A indicated that three to five amino acids are removed from the C terminus of the cytokine. Meprin A and meprin B cleaved IL-6 with micromolar affinities (Km of 4.7 and 12.0 μm, respectively) and with high efficiencies (kcat/Km of 0.2 and 2.5 (m−1/s−1) × 106, respectively). These efficiency constants are among the highest for known meprin substrates. Madin-Darby canine kidney cells transiently transfected with meprin α or meprin β constructs also cleave exogenous IL-6. Both human and murine IL-6 cleaved by meprin A or B are inactivated, as demonstrated by their decreased capability to stimulate proliferation of B9 cells. These results are consistent with the proposition that one function of meprin metalloproteases is to modulate inflammation by inactivating IL-6. PMID:24474695

  10. A study of severance taxes on crude oil and natural gas: The irreversibility of taxation

    NASA Astrophysics Data System (ADS)

    Brandly, Mark L.

    This dissertation examines the institution of severance taxes. An explanation of the property rights allocations in the petroleum industry provides the foundation for discussing the incentive structure of the industry. This explanation concludes that the severance tax burden on the supply side of the industry is born by oil producers and royalty owners. A history of national and state severance taxes in the United States is provided. The literature on the justifications for severance taxes and the economic studies that are relevant to the issue of the tax effect on oil output is reviewed. This review shows that an important implication of severance taxes, the fact that the output effect of such taxes is at least partially irreversible, has been overlooked. A mathematical model is constructed that demonstrates the relationships between output, the sellers' price, the buyers' price, excess burden, the consumers' tax burden, the producers' tax burden, and the price elasticities of supply and demand. It is then demonstrated that the appropriate framework for analyzing severance taxes includes an upward sloping supply curve and a completely elastic demand curve. Another mathematical model shows the effect that a severance tax has on the output decision given different income tax situations. A review of the industry procedures for abandoning wells is followed by a theoretical argument that severance taxes are irreversible to some degree. When a well is abandoned, due to a severance tax, the well is plugged with cement. The costs of reentering such a well are large relative to the potential profits to be derived from such a decision. Eliminating the severance tax does not provide the incentive needed to reenter and produce an abandoned well. An empirical examination of the Kansas severance tax imposed in 1983 compares the present value of an abandoned well with the costs of reentering such a well. This comparison leads to the conclusion that, generally, a well that was

  11. Typical pure nonequilibrium steady states and irreversibility for quantum transport.

    PubMed

    Monnai, Takaaki; Yuasa, Kazuya

    2016-07-01

    It is known that each single typical pure state in an energy shell of a large isolated quantum system well represents a thermal equilibrium state of the system. We show that such typicality holds also for nonequilibrium steady states (NESS's). We consider a small quantum system coupled to multiple infinite reservoirs. In the long run, the total system reaches a unique NESS. We identify a large Hilbert space from which pure states of the system are to be sampled randomly and show that the typical pure states well describe the NESS. We also point out that the irreversible relaxation to the unique NESS is important to the typicality of the pure NESS's.

  12. Typical pure nonequilibrium steady states and irreversibility for quantum transport

    NASA Astrophysics Data System (ADS)

    Monnai, Takaaki; Yuasa, Kazuya

    2016-07-01

    It is known that each single typical pure state in an energy shell of a large isolated quantum system well represents a thermal equilibrium state of the system. We show that such typicality holds also for nonequilibrium steady states (NESS's). We consider a small quantum system coupled to multiple infinite reservoirs. In the long run, the total system reaches a unique NESS. We identify a large Hilbert space from which pure states of the system are to be sampled randomly and show that the typical pure states well describe the NESS. We also point out that the irreversible relaxation to the unique NESS is important to the typicality of the pure NESS's.

  13. Discovery of a selective irreversible BMX inhibitor for prostate cancer.

    PubMed

    Liu, Feiyang; Zhang, Xin; Weisberg, Ellen; Chen, Sen; Hur, Wooyoung; Wu, Hong; Zhao, Zheng; Wang, Wenchao; Mao, Mao; Cai, Changmeng; Simon, Nicholas I; Sanda, Takaomi; Wang, Jinhua; Look, A Thomas; Griffin, James D; Balk, Steven P; Liu, Qingsong; Gray, Nathanael S

    2013-07-19

    BMX is a member of the TEC family of nonreceptor tyrosine kinases. We have used structure-based drug design in conjunction with kinome profiling to develop a potent, selective, and irreversible BMX kinase inhibitor, BMX-IN-1, which covalently modifies Cys496. BMX-IN-1 inhibits the proliferation of Tel-BMX-transformed Ba/F3 cells at two digit nanomolar concentrations but requires single digit micromolar concentrations to inhibit the proliferation of prostate cancer cell lines. Using a combinatorial kinase inhibitor screening strategy, we discovered that the allosteric Akt inhibitor, MK2206, is able to potentiate BMX inhibitor's antiproliferation efficacy against prostate cancer cells. PMID:23594111

  14. Expedited Record Base Fabrication Using an Irreversible Hydrocolloid Cast.

    PubMed

    Oh, Won-suk; Park, Ju-mi

    2016-04-01

    The registration of a maxillomandibular relationship requires additional clinical and laboratory procedures when the mouth presents with loss of occlusal support. This procedure can be a challenge for a patient who needs urgent care or resides in a remote area. This article describes a procedure for expediting the mounting of a master cast for the fabrication of a maxillary immediate complete denture. The technique presented describes the use of a silicone record base made on an irreversible hydrocolloid cast generated from the final impression.

  15. Intrinsic irreversibility limits the efficiency of multidimensional molecular motors.

    PubMed

    Jack, M W; Tumlin, C

    2016-05-01

    We consider the efficiency limits of Brownian motors able to extract work from the temperature difference between reservoirs or from external thermodynamic forces. These systems can operate in a variety of modes, including as isothermal engines, heat engines, refrigerators, and heat pumps. We derive analytical results showing that certain classes of multidimensional Brownian motor, including the Smoluchowski-Feynman ratchet, are unable to attain perfect efficiency (Carnot efficiency for heat engines). This demonstrates the presence of intrinsic irreversibilities in their operating mechanism. We present numerical simulations showing that in some cases the loss process that limits efficiency is associated with vortices in the probability current.

  16. Microscopic time-reversibility and macroscopic irreversibility: Still a paradox

    SciTech Connect

    Posch, H.A.; Dellago, Ch.; Hoover, W.G.; Kum, O. |

    1995-09-13

    Microscopic time reversibility and macroscopic irreversibility are a paradoxical combination. This was first observed by J. Loschmidt in 1876 and was explained, for conservative systems, by L. Boltzmann the following year. Both these features are also present in modern simulations of classic many-body systems in steady nonequilibrium states. We illustrate them here for the simplest possible models, a continuous one-dimensional model of field-driven diffusion, the so-called driven Lorentz gas or Galton Board, and an ergodic time reversible dissipative map.

  17. Intrinsic irreversibility limits the efficiency of multidimensional molecular motors

    NASA Astrophysics Data System (ADS)

    Jack, M. W.; Tumlin, C.

    2016-05-01

    We consider the efficiency limits of Brownian motors able to extract work from the temperature difference between reservoirs or from external thermodynamic forces. These systems can operate in a variety of modes, including as isothermal engines, heat engines, refrigerators, and heat pumps. We derive analytical results showing that certain classes of multidimensional Brownian motor, including the Smoluchowski-Feynman ratchet, are unable to attain perfect efficiency (Carnot efficiency for heat engines). This demonstrates the presence of intrinsic irreversibilities in their operating mechanism. We present numerical simulations showing that in some cases the loss process that limits efficiency is associated with vortices in the probability current.

  18. Intrinsic irreversibility limits the efficiency of multidimensional molecular motors.

    PubMed

    Jack, M W; Tumlin, C

    2016-05-01

    We consider the efficiency limits of Brownian motors able to extract work from the temperature difference between reservoirs or from external thermodynamic forces. These systems can operate in a variety of modes, including as isothermal engines, heat engines, refrigerators, and heat pumps. We derive analytical results showing that certain classes of multidimensional Brownian motor, including the Smoluchowski-Feynman ratchet, are unable to attain perfect efficiency (Carnot efficiency for heat engines). This demonstrates the presence of intrinsic irreversibilities in their operating mechanism. We present numerical simulations showing that in some cases the loss process that limits efficiency is associated with vortices in the probability current. PMID:27300832

  19. Quantum Chaos, Irreversible Classical Dynamics, and Random Matrix Theory

    NASA Astrophysics Data System (ADS)

    Andreev, A. V.; Agam, O.; Simons, B. D.; Altshuler, B. L.

    1996-05-01

    The Bohigas-Giannoni-Schmit conjecture stating that the statistical spectral properties of systems which are chaotic in their classical limit coincide with random matrix theory (RMT) is proved. A new semiclassical field theory for individual chaotic systems is constructed in the framework of a nonlinear σ model. The low lying modes are shown to be associated with the Perron-Frobenius (PF) spectrum of the underlying irreversible classical dynamics. It is shown that the existence of a gap in the PF spectrum results in RMT behavior. Moreover, our formalism offers a way of calculating system specific corrections beyond RMT.

  20. Independent evolution of transcriptional inactivation on sex chromosomes in birds and mammals.

    PubMed

    Livernois, Alexandra M; Waters, Shafagh A; Deakin, Janine E; Marshall Graves, Jennifer A; Waters, Paul D

    2013-01-01

    X chromosome inactivation in eutherian mammals has been thought to be tightly controlled, as expected from a mechanism that compensates for the different dosage of X-borne genes in XX females and XY males. However, many X genes escape inactivation in humans, inactivation of the X in marsupials is partial, and the unrelated sex chromosomes of monotreme mammals have incomplete and gene-specific inactivation of X-linked genes. The bird ZW sex chromosome system represents a third independently evolved amniote sex chromosome system with dosage compensation, albeit partial and gene-specific, via an unknown mechanism (i.e. upregulation of the single Z in females, down regulation of one or both Zs in males, or a combination). We used RNA-fluorescent in situ hybridization (RNA-FISH) to demonstrate, on individual fibroblast cells, inactivation of 11 genes on the chicken Z and 28 genes on the X chromosomes of platypus. Each gene displayed a reproducible frequency of 1Z/1X-active and 2Z/2X-active cells in the homogametic sex. Our results indicate that the probability of inactivation is controlled on a gene-by-gene basis (or small domains) on the chicken Z and platypus X chromosomes. This regulatory mechanism must have been exapted independently to the non-homologous sex chromosomes in birds and mammals in response to an over-expressed Z or X in the homogametic sex, highlighting the universal importance that (at least partial) silencing plays in the evolution on amniote dosage compensation and, therefore, the differentiation of sex chromosomes. PMID:23874231

  1. Using silicone impression material and acrylic resin to fabricate remount casts for removable partial dentures.

    PubMed

    Hsu, Yung-tsung; Farmer, John B

    2005-06-01

    A technique is described for fabrication of a remount cast for a removable partial denture. This procedure consists of filling the occlusal/incisal third with acrylic resin and injecting polyvinylsiloxane impression material into the irreversible hydrocolloid impression. This technique provides a simple method for making a remount cast and enables the clinician to remove and easily place the partial denture on the cast during occlusal refinement procedures without damage to the removable partial denture or the remount cast.

  2. Engineered Covalent Inactivation of TFIIH-Kinase Reveals an Elongation Checkpoint and Results in Widespread mRNA Stabilization.

    PubMed

    Rodríguez-Molina, Juan B; Tseng, Sandra C; Simonett, Shane P; Taunton, Jack; Ansari, Aseem Z

    2016-08-01

    During transcription initiation, the TFIIH-kinase Kin28/Cdk7 marks RNA polymerase II (Pol II) by phosphorylating the C-terminal domain (CTD) of its largest subunit. Here we describe a structure-guided chemical approach to covalently and specifically inactivate Kin28 kinase activity in vivo. This method of irreversible inactivation recapitulates both the lethal phenotype and the key molecular signatures that result from genetically disrupting Kin28 function in vivo. Inactivating Kin28 impacts promoter release to differing degrees and reveals a "checkpoint" during the transition to productive elongation. While promoter-proximal pausing is not observed in budding yeast, inhibition of Kin28 attenuates elongation-licensing signals, resulting in Pol II accumulation at the +2 nucleosome and reduced transition to productive elongation. Furthermore, upon inhibition, global stabilization of mRNA masks different degrees of reduction in nascent transcription. This study resolves long-standing controversies on the role of Kin28 in transcription and provides a rational approach to irreversibly inhibit other kinases in vivo. PMID:27477907

  3. Inactivation of enzymes and oxidative modification of proteins by stimulated neutrophils.

    PubMed

    Oliver, C N

    1987-02-15

    Differentiated, stimulated HL-60 cells and freshly isolated, stimulated neutrophils inactivate glutamine synthetase (L-glutamate:ammonia ligase (ADP-forming), EC 6.3.1.2) either inside or outside of Escherichia coli. Stimulated neutrophils also inactivate at least four endogenous enzymes which are inactivated by mixed-function oxidation (MFO) systems in vitro (L. Fucci, C.N. Oliver, M.J. Coon, and E.R. Stadtman (1983) Proc. Natl. Acad. Sci. USA 80, 1521-1525). The inactivation of glutamine synthetase by stimulated neutrophils exhibits characteristics similar to those previously described using both enzymic and nonenzymic MFO systems (R.L. Levine, C.N. Oliver, R.M. Fulks, and E.R. Stadtman (1981) Proc. Natl. Acad. Sci. USA 78, 2120-2124). Although the reaction occurs in the absence of Fe(III), it is stimulated by added Fe (III). Inactivation required molecular oxygen and is partially inhibited by Mn(II), catalase, superoxide dismutase, and metal chelators, ethylenediaminetetraacetic acid and o-phenanthroline. Both the kinetics and the extent of glutamine synthetase inactivation differ when neutrophils are stimulated with phorbol esters compared with formylated peptides. Glutamine synthetase inactivation catalyzed by MFO systems is accompanied by the formation of protein carbonyl derivatives which form stable hydrazones when treated with 2,4-dinitrophenylhydrazine. Multiple carbonyl derivatives are formed in the soluble protein fraction of stimulated neutrophils and these derivatives collectively exhibit an absorbance spectrum similar to that of glutamine synthetase inactivated by liver microsomal cytochrome P-450 MFO system (K. Nakamura, C.N. Oliver, and E.R. Stadtman (1985) Arch. Biochem. Biophys. 240, 319-329).

  4. Cinnamic acid 4-hydroxylase mechanism-based inactivation by psoralen derivatives: cloning and characterization of a C4H from a psoralen producing plant-Ruta graveolens-exhibiting low sensitivity to psoralen inactivation.

    PubMed

    Gravot, Antoine; Larbat, Romain; Hehn, Alain; Lièvre, Karine; Gontier, Eric; Goergen, Jean Louis; Bourgaud, Frédéric

    2004-02-01

    Cinnamate 4-hydroxylase (C4H, EC 1.14.13.11) complete cDNA was cloned from the leaves of Ruta graveolens, a psoralen producing plant. The recombinant enzyme (classified CYP73A32) was expressed in Saccharomyces cerevisiae. Mechanism-based inactivation was investigated using various psoralen derivatives. Only psoralen and 8-methoxypsoralen were found to inactivate C4H. The inactivation was dependent on the presence of NADPH, time of pre-incubation, and inhibitor concentration. Inactivation stoichiometry was 0.9 (+/-0.2) for CYP73A1 and 1.1 (+/-0.2) for CYP73A32. SDS-PAGE analysis demonstrated that [3H]psoralen was irreversibly bound to the C4H apoprotein. K(i) and k(inact) for psoralen and 8-methoxypsoralen inactivation on the two C4H revealed a lower sensitivity for CYP73A32 compared to CYP73A1. Inactivation kinetics were also determined for CYP73A10, a C4H from another furocoumarin-producing plant, Petroselinum crispum. This enzyme was found to behave like CYP73A32, with a weak sensitivity to psoralen and 8-MOP inactivation. Cinnamic acid hydroxylation is a key step in the biosynthesis of phenylpropanoid compounds, psoralen derivatives included. Our results suggest a possible evolution of R. graveolens and P. crispum C4H that might tolerate substantial levels of psoralen derivatives in the cytoplasmic compartment without a depletive effect on C4H and the general phenylpropanoid metabolism.

  5. Inactivation and injury assessment of Escherichia coli during solar and photocatalytic disinfection in LDPE bags.

    PubMed

    Dunlop, P S M; Ciavola, M; Rizzo, L; Byrne, J A

    2011-11-01

    Solar disinfection (SODIS) of Escherichia coli suspensions in low-density polyethylene bag reactors was investigated as a low-cost disinfection method suitable for application in developing countries. The efficiency of a range of SODIS reactor configurations was examined (single skin (SS), double skin, black-backed single skin, silver-backed single skin (SBSS) and composite-backed single skin) using E. coli suspended in model and real surface water. Titanium dioxide was added to the reactors to improve the efficiency of the SODIS process. The effect of turbidity was also assessed. In addition to viable counts, E. coli injury was characterised through spread-plate analysis using selective and non-selective media. The optimal reactor configuration was determined to be the SBSS bag (t(50)=9.0min) demonstrating the importance of UVA photons, as opposed to infrared in the SODIS disinfection mechanism. Complete inactivation (6.5-log) was achieved in the presence of turbidity (50NTU) using the SBSS bag within 180min simulated solar exposure. The addition of titanium dioxide (0.025gL(-1)) significantly enhanced E. coli inactivation in the SS reactor, with 6-log inactivation observed within 90min simulated solar exposure. During the early stages of both SODIS and photocatalytic disinfection, injured E. coli were detected; however, irreversible injury was caused and re-growth was not observed. Experiments under solar conditions were undertaken with total inactivation (6.5-log) observed in the SS reactor within 240min, incomplete inactivation (4-log) was observed in SODIS bottles exposed to the same solar conditions.

  6. Inactivation of Giardia lamblia cysts with ozone.

    PubMed Central

    Wickramanayake, G B; Rubin, A J; Sproul, O J

    1984-01-01

    Giardia lamblia cysts were inactivated in water with ozone at pH 7.0 and 5 and 25 degrees C. The concentration-time products for 99% inactivation were 0.53 and 0.17 mg-min/liter at 5 and 25 degrees C, respectively. These products were significantly lower than those reported for chlorine. PMID:6497374

  7. Carnot's cycle for small systems: Irreversibility and cost of operations

    NASA Astrophysics Data System (ADS)

    Sekimoto, Ken; Takagi, Fumiko; Hondou, Tsuyoshi

    2000-12-01

    In the thermodynamic limit, the existence of a maximal efficiency of energy conversion attainable by a Carnot cycle consisting of quasistatic isothermal and adiabatic processes precludes the existence of a perpetual machine of the second kind, whose cycles yield positive work in an isothermal environment. We employ the recently developed framework of the energetics of stochastic processes (called ``stochastic energetics'') to reanalyze the Carnot cycle in detail, taking account of fluctuations, without taking the thermodynamic limit. We find that in this nonmacroscopic situation both processes of connection to and disconnection from heat baths and adiabatic processes that cause distortion of the energy distribution are sources of inevitable irreversibility within the cycle. Also, the so-called null-recurrence property of the cumulative efficiency of energy conversion over many cycles and the irreversible property of isolated, purely mechanical processes under external ``macroscopic'' operations are discussed in relation to the impossibility of a perpetual machine, or Maxwell's demon. This analysis may serve as the basis for the design and analysis of mesoscopic energy converters in the near future.

  8. Irreversible entropy model for damage diagnosis in resistors

    SciTech Connect

    Cuadras, Angel Crisóstomo, Javier; Ovejas, Victoria J.; Quilez, Marcos

    2015-10-28

    We propose a method to characterize electrical resistor damage based on entropy measurements. Irreversible entropy and the rate at which it is generated are more convenient parameters than resistance for describing damage because they are essentially positive in virtue of the second law of thermodynamics, whereas resistance may increase or decrease depending on the degradation mechanism. Commercial resistors were tested in order to characterize the damage induced by power surges. Resistors were biased with constant and pulsed voltage signals, leading to power dissipation in the range of 4–8 W, which is well above the 0.25 W nominal power to initiate failure. Entropy was inferred from the added power and temperature evolution. A model is proposed to understand the relationship among resistance, entropy, and damage. The power surge dissipates into heat (Joule effect) and damages the resistor. The results show a correlation between entropy generation rate and resistor failure. We conclude that damage can be conveniently assessed from irreversible entropy generation. Our results for resistors can be easily extrapolated to other systems or machines that can be modeled based on their resistance.

  9. Irreversible Langevin samplers and variance reduction: a large deviations approach

    NASA Astrophysics Data System (ADS)

    Rey-Bellet, Luc; Spiliopoulos, Konstantinos

    2015-07-01

    In order to sample from a given target distribution (often of Gibbs type), the Monte Carlo Markov chain method consists of constructing an ergodic Markov process whose invariant measure is the target distribution. By sampling the Markov process one can then compute, approximately, expectations of observables with respect to the target distribution. Often the Markov processes used in practice are time-reversible (i.e. they satisfy detailed balance), but our main goal here is to assess and quantify how the addition of a non-reversible part to the process can be used to improve the sampling properties. We focus on the diffusion setting (overdamped Langevin equations) where the drift consists of a gradient vector field as well as another drift which breaks the reversibility of the process but is chosen to preserve the Gibbs measure. In this paper we use the large deviation rate function for the empirical measure as a tool to analyze the speed of convergence to the invariant measure. We show that the addition of an irreversible drift leads to a larger rate function and it strictly improves the speed of convergence of ergodic average for (generic smooth) observables. We also deduce from this result that the asymptotic variance decreases under the addition of the irreversible drift and we give an explicit characterization of the observables whose variance is not reduced reduced, in terms of a nonlinear Poisson equation. Our theoretical results are illustrated and supplemented by numerical simulations.

  10. Evidence of irreversible CO2 intercalation in montmorillonite

    SciTech Connect

    Romanov, V

    2013-02-13

    Mitigation of the global climate change via sequestration of anthropogenic carbon dioxide (CO2) in geologic formations requires assessment of the reservoir storage capacity and cap rock seal integrity. The typical cap rock is shale or mudstone rich in clay minerals that may significantly affect the effectiveness of the CO2 trapping. Specific objectives of this study were to conduct experimental investigation into the processes associated with CO2 and H2O trapped in swelling clay, namely, Wyoming and Texas montmorillonite powder. Combined (same-sample) multi-technique data ? manometric sorption isotherm hysteresis, diffuse reflectance infrared spectroscopy ?trapped CO2? fingerprints, irreversible X-ray diffraction patterns for the clay interlayer in intermediate hydration state, and HF acid digestion resulting in formation of non-extractable F:CO2 adducts ? corroborate a hypothesis that carbon dioxide molecules can be irreversibly trapped via anomalous extreme confinement in the galleries associated with montmorillonite interlayer, which may result in formation of carbonates in the longer term. Validation on Arizona montmorillonite lumps substantiated the evidence that such processes may occur in natural clay deposits but possibly on a different scale and at a different rate.

  11. Irreversibility in physics stemming from unpredictable symbol-handling agents

    NASA Astrophysics Data System (ADS)

    Myers, John M.; Madjid, F. Hadi

    2016-05-01

    The basic equations of physics involve a time variable t and are invariant under the transformation t --> -t. This invariance at first sight appears to impose time reversibility as a principle of physics, in conflict with thermodynamics. But equations written on the blackboard are not the whole story in physics. In prior work we sharpened a distinction obscured in today's theoretical physics, the distinction between obtaining evidence from experiments on the laboratory bench and explaining that evidence in mathematical symbols on the blackboard. The sharp distinction rests on a proof within the mathematics of quantum theory that no amount of evidence, represented in quantum theory in terms of probabilities, can uniquely determine its explanation in terms of wave functions and linear operators. Building on the proof we show here a role in physics for unpredictable symbol-handling agents acting both at the blackboard and at the workbench, communicating back and forth by means of transmitted symbols. Because of their unpredictability, symbol-handling agents introduce a heretofore overlooked source of irreversibility into physics, even when the equations they write on the blackboard are invariant under t --> -t. Widening the scope of descriptions admissible to physics to include the agents and the symbols that link theory to experiments opens up a new source of time-irreversibility in physics.

  12. When an adiabatic irreversible expansion or compression becomes reversible

    NASA Astrophysics Data System (ADS)

    Anacleto, Joaquim; Ferreira, J. M.; Soares, A. A.

    2009-05-01

    This paper aims to contribute to a better understanding of the concepts of a reversible process and entropy. For this purpose, an adiabatic irreversible expansion or compression is analysed, by considering that an ideal gas is expanded (compressed), from an initial pressure Pi to a final pressure Pf, by being placed in contact with a set of N work reservoirs with pressures decreasing (increasing) in a geometric or arithmetic progression. The gas entropy change ΔS is evaluated and it is clearly shown that ΔS > 0 for any finite N, but as the number of work reservoirs goes to infinity the entropy change goes to zero, i.e. the process becomes reversible. Additionally, this work draws attention to the work reservoir concept, which is virtually ignored in the literature, and to its analogy with the commonly used heat reservoir concept. Finally, it complements and reinforces an earlier study dealing with irreversible cooling or heating so that the synergy created by the two studies is important from both theoretical and educational standpoints.

  13. From Maximum Entropy Models to Non-Stationarity and Irreversibility

    NASA Astrophysics Data System (ADS)

    Cofre, Rodrigo; Cessac, Bruno; Maldonado, Cesar

    The maximum entropy distribution can be obtained from a variational principle. This is important as a matter of principle and for the purpose of finding approximate solutions. One can exploit this fact to obtain relevant information about the underlying stochastic process. We report here in recent progress in three aspects to this approach.1- Biological systems are expected to show some degree of irreversibility in time. Based on the transfer matrix technique to find the spatio-temporal maximum entropy distribution, we build a framework to quantify the degree of irreversibility of any maximum entropy distribution.2- The maximum entropy solution is characterized by a functional called Gibbs free energy (solution of the variational principle). The Legendre transformation of this functional is the rate function, which controls the speed of convergence of empirical averages to their ergodic mean. We show how the correct description of this functional is determinant for a more rigorous characterization of first and higher order phase transitions.3- We assess the impact of a weak time-dependent external stimulus on the collective statistics of spiking neuronal networks. We show how to evaluate this impact on any higher order spatio-temporal correlation. RC supported by ERC advanced Grant ``Bridges'', BC: KEOPS ANR-CONICYT, Renvision and CM: CONICYT-FONDECYT No. 3140572.

  14. Fluctuations in the number of irreversibly adsorbed particles

    NASA Astrophysics Data System (ADS)

    Adamczyk, Zbigniew; Szyk-Warszyńska, Lilianna; Siwek, B.; Weroński, P.

    2000-12-01

    Fluctuations in the number of colloid particles adsorbed irreversibly under pure diffusion transport conditions were determined as a function of surface density and ionic strength of the suspension. The experiments were carried out for monodisperse polystyrene latex particles of micrometer size range adsorbing irreversibly at mica surface. The surface density of adsorbed particles at various areas was determined using the direct microscope observation method. A new experimental cell was used enabling in situ observations of particles adsorption under conditions of negligible gravity effects. It was found that the particle density fluctuations for high ionic strength were in a good agreement with the theoretical results derived from the random sequential adsorption (RSA) model. Also, the theoretical results stemming from the equilibrium scaled particle theory reflected the experimental data satisfactorily. For lower ionic strength a deviation from the hard sphere behavior was experimentally demonstrated. This effect due to the repulsive electrostatic interactions was interpreted in terms of the effective hard particle concept. The universal dependence of variance on particle density obtained in this way was found in a good agreement with the RSA model for all ionic strength. These results proved that fluctuations in particle density of monolayer formed under diffusional conditions differ fundamentally from these obtained under ballistic transport conditions.

  15. Molecular control of irreversible bistability during trypanosome developmental commitment

    PubMed Central

    Domingo-Sananes, Maria Rosa; Szöőr, Balazs; Ferguson, Michael A.J.

    2015-01-01

    The life cycle of Trypanosoma brucei involves developmental transitions that allow survival, proliferation, and transmission of these parasites. One of these, the differentiation of growth-arrested stumpy forms in the mammalian blood into insect-stage procyclic forms, can be induced synchronously in vitro with cis-aconitate. Here, we show that this transition is an irreversible bistable switch, and we map the point of commitment to differentiation after exposure to cis-aconitate. This irreversibility implies that positive feedback mechanisms operate to allow commitment (i.e., the establishment of “memory” of exposure to the differentiation signal). Using the reversible translational inhibitor cycloheximide, we show that this signal memory requires new protein synthesis. We further performed stable isotope labeling by amino acids in cell culture to analyze synchronized parasite populations, establishing the protein and phosphorylation profile of parasites pre- and postcommitment, thereby defining the “commitment proteome.” Functional interrogation of this data set identified Nek-related kinase as the first-discovered protein kinase controlling the initiation of differentiation to procyclic forms. PMID:26483558

  16. Linear Dimensional Stability of Irreversible Hydrocolloid Materials Over Time.

    PubMed

    Garrofé, Analía B; Ferrari, Beatriz A; Picca, Mariana; Kaplan, Andrea E

    2015-12-01

    The aim of this study was to evaluate the linear dimensional stability of different irreversible hydrocolloid materials over time. A metal mold was designed with custom trays made of thermoplastic sheets (Sabilex, sheets 0.125 mm thick). Perforations were made in order to improve retention of the material. Five impressions were taken with each of the following: Kromopan 100 (LASCOD) [AlKr], which has dimensional stability of 100 hours, and Phase Plus (ZHERMACK) [AlPh], which has dimensional stability of 48 hours. Standardized digital photographs were taken at different time intervals (0, 15, 30, 45, 60, 120 minutes; 12, 24 and 96 hours), using an "ad-hoc" device. The images were analyzed with software (UTHSCSA Image Tool) by measuring the distance between intersection of the lines previously made at the top of the mold. The results were analyzed by ANOVA for repeated measures. Initial and final values were (mean and standard deviation): AlKr: 16.44 (0.22) and 16.34 (0.11), AlPh: 16.40 (0.06) and 16.18 (0.06). Statistical evaluation showed significant effect of material and time factors. Under the conditions in this study, time significantly affects the linear dimensional stability of irreversible hydrocolloid materials. PMID:27095627

  17. Essays on oil price volatility and irreversible investment

    NASA Astrophysics Data System (ADS)

    Pastor, Daniel J.

    In chapter 1, we provide an extensive and systematic evaluation of the relative forecasting performance of several models for the volatility of daily spot crude oil prices. Empirical research over the past decades has uncovered significant gains in forecasting performance of Markov Switching GARCH models over GARCH models for the volatility of financial assets and crude oil futures. We find that, for spot oil price returns, non-switching models perform better in the short run, whereas switching models tend to do better at longer horizons. In chapter 2, I investigate the impact of volatility on firms' irreversible investment decisions using real options theory. Cost incurred in oil drilling is considered sunk cost, thus irreversible. I collect detailed data on onshore, development oil well drilling on the North Slope of Alaska from 2003 to 2014. Volatility is modeled by constructing GARCH, EGARCH, and GJR-GARCH forecasts based on monthly real oil prices, and realized volatility from 5-minute intraday returns of oil futures prices. Using a duration model, I show that oil price volatility generally has a negative relationship with the hazard rate of drilling an oil well both when aggregating all the fields, and in individual fields.

  18. Linear Dimensional Stability of Irreversible Hydrocolloid Materials Over Time.

    PubMed

    Garrofé, Analía B; Ferrari, Beatriz A; Picca, Mariana; Kaplan, Andrea E

    2015-12-01

    The aim of this study was to evaluate the linear dimensional stability of different irreversible hydrocolloid materials over time. A metal mold was designed with custom trays made of thermoplastic sheets (Sabilex, sheets 0.125 mm thick). Perforations were made in order to improve retention of the material. Five impressions were taken with each of the following: Kromopan 100 (LASCOD) [AlKr], which has dimensional stability of 100 hours, and Phase Plus (ZHERMACK) [AlPh], which has dimensional stability of 48 hours. Standardized digital photographs were taken at different time intervals (0, 15, 30, 45, 60, 120 minutes; 12, 24 and 96 hours), using an "ad-hoc" device. The images were analyzed with software (UTHSCSA Image Tool) by measuring the distance between intersection of the lines previously made at the top of the mold. The results were analyzed by ANOVA for repeated measures. Initial and final values were (mean and standard deviation): AlKr: 16.44 (0.22) and 16.34 (0.11), AlPh: 16.40 (0.06) and 16.18 (0.06). Statistical evaluation showed significant effect of material and time factors. Under the conditions in this study, time significantly affects the linear dimensional stability of irreversible hydrocolloid materials.

  19. Voter model with arbitrary degree dependence: clout, confidence and irreversibility

    NASA Astrophysics Data System (ADS)

    Fotouhi, Babak; Rabbat, Michael G.

    2014-03-01

    The voter model is widely used to model opinion dynamics in society. In this paper, we propose three modifications to incorporate heterogeneity into the model. We address the corresponding oversimplifications of the conventional voter model which are unrealistic. We first consider the voter model with popularity bias. The influence of each node on its neighbors depends on its degree. We find the consensus probabilities and expected consensus times for each of the states. We also find the fixation probability, which is the probability that a single node whose state differs from every other node imposes its state on the entire system. In addition, we find the expected fixation time. Then two other extensions to the model are proposed and the motivations behind them are discussed. The first one is confidence, where in addition to the states of neighbors, nodes take their own state into account at each update. We repeat the calculations for the augmented model and investigate the effects of adding confidence to the model. The second proposed extension is irreversibility, where one of the states is given the property that once nodes adopt it, they cannot switch back. This is motivated by applications where, agents take an irreversible action such as seeing a movie, purchasing a music album online, or buying a new product. The dynamics of densities, fixation times and consensus times are obtained.

  20. Evidence for the Degradation of Nicotinamide Adenine Dinucleotide Phosphate-Dependent Glutamate Dehydrogenase of Candida utilis During Rapid Enzyme Inactivation

    PubMed Central

    Hemmings, Brian A.

    1978-01-01

    The nicotinamide adenine dinucleotide phosphate-dependent glutamate dehydrogenase (NADP-GDH) from the food yeast Candida utilis was found to be rapidly inactivated when cultures were starved of a carbon source. The addition of glutamate or alanine to the starvation medium stimulated the rate of inactivation. Loss of enzyme activity was irreversible since the reappearance of enzyme activity, following the addition of glucose to carbon-starved cultures, was blocked by cycloheximide. A specific rabbit antibody was prepared against the NADP-GDH from C. utilis and used to quantitate the enzyme during inactivation promoted by carbon starvation. The amount of precipitable antigenic material paralleled the rapid decrease of enzyme activity observed after transition of cells from NH4+-glucose to glutamate medium. No additional small-molecular-weight protein was precipitated by the antibody as a result of the inactivation, suggesting that the enzyme is considerably altered during the primary steps of the inactivation process. Analysis by immunoprecipitation of the reappearance of enzyme activity after enzyme inactivation showed that increase of NADP-GDH activity was almost totally due to de novo synthesis, ruling out the possibility that enzyme activity modulation is achieved by reversible covalent modification. Enzyme degradation was also measured during steady-state growth and other changes in nitrogen and carbon status of the culture media. In all instances so far estimated, the enzyme was found to be very stable and not normally subject to high rates of degradation. Therefore, the possibility that inactivation was caused by a change in the ratio of synthesis to degradation can be excluded. Images PMID:24041

  1. Photodynamic Inactivation of Mammalian Viruses and Bacteriophages

    PubMed Central

    Costa, Liliana; Faustino, Maria Amparo F.; Neves, Maria Graça P. M. S.; Cunha, Ângela; Almeida, Adelaide

    2012-01-01

    Photodynamic inactivation (PDI) has been used to inactivate microorganisms through the use of photosensitizers. The inactivation of mammalian viruses and bacteriophages by photosensitization has been applied with success since the first decades of the last century. Due to the fact that mammalian viruses are known to pose a threat to public health and that bacteriophages are frequently used as models of mammalian viruses, it is important to know and understand the mechanisms and photodynamic procedures involved in their photoinactivation. The aim of this review is to (i) summarize the main approaches developed until now for the photodynamic inactivation of bacteriophages and mammalian viruses and, (ii) discuss and compare the present state of the art of mammalian viruses PDI with phage photoinactivation, with special focus on the most relevant mechanisms, molecular targets and factors affecting the viral inactivation process. PMID:22852040

  2. Photodynamic inactivation of mammalian viruses and bacteriophages.

    PubMed

    Costa, Liliana; Faustino, Maria Amparo F; Neves, Maria Graça P M S; Cunha, Angela; Almeida, Adelaide

    2012-07-01

    Photodynamic inactivation (PDI) has been used to inactivate microorganisms through the use of photosensitizers. The inactivation of mammalian viruses and bacteriophages by photosensitization has been applied with success since the first decades of the last century. Due to the fact that mammalian viruses are known to pose a threat to public health and that bacteriophages are frequently used as models of mammalian viruses, it is important to know and understand the mechanisms and photodynamic procedures involved in their photoinactivation. The aim of this review is to (i) summarize the main approaches developed until now for the photodynamic inactivation of bacteriophages and mammalian viruses and, (ii) discuss and compare the present state of the art of mammalian viruses PDI with phage photoinactivation, with special focus on the most relevant mechanisms, molecular targets and factors affecting the viral inactivation process.

  3. Inactivation of Ascaris suum eggs by ammonia.

    PubMed

    Pecson, Brian M; Nelson, Kara L

    2005-10-15

    Uncharged ammonia is known to cause inactivation of a number of wastewater pathogens, but its effect on Ascaris eggs has never been isolated or quantified. The objectives of this research were to determine the conditions under which ammonia inactivates eggs of the swine Ascaris species, Ascaris suum, and to quantify the impact of ammonia on the U.S. EPA's requirements for alkaline treatment to produce Class A sludge. Eggs were incubated in controlled, laboratory solutions such that the effects of ammonia concentration and speciation, pH, and temperature could be separated. With a 24-h incubation, the inactivation at all pH levels (range 7-11) was not statistically different in the absence of ammonia. The presence of ammonia (0-1000 ppm as N) significantly increased Ascaris egg inactivation at pH 9 and 11, and the ovicidal effect was directly related to the concentration of the uncharged NH3 species. Increasing temperatures (32-52 degrees C) caused increased inactivation at all pH levels and ammonia concentrations. The current EPA treatment requirements to produce Class A biosolids by alkaline treatment have temperature, pH, and time requirements, but do not account for the effectof differences in ammonia concentration on inactivation. To illustrate the potential savings in temperature and pH that could be achieved when accounting for ammonia inactivation, the combinations of ammonia concentration, temperature, and pH neededto achieve 99% inactivation after 72 h were determined. The presence of ammonia at concentrations encountered in sludges and feces (up to 8000 ppm as N) allowed for 99% egg inactivation to be achieved at temperatures up to 14 degrees C lower than ammonia-free controls. Thus, environmentally relevant concentrations of ammonia may significantly increase the rate of Ascaris egg inactivation during alkaline stabilization.

  4. Inactivation of peptidylglycine α-hydroxylating monooxygenase by cinnamic acid analogs.

    PubMed

    McIntyre, Neil R; Lowe, Edward W; Battistini, Matthew R; Leahy, James W; Merkler, David J

    2016-08-01

    Peptidylglycine α-amidating monooxygenase (PAM) is a bifunctional enzyme that catalyzes the final reaction in the maturation of α-amidated peptide hormones. Peptidylglycine α-hydroxylating monooxygenase (PHM) is the PAM domain responsible for the copper-, ascorbate- and O2-dependent hydroxylation of a glycine-extended peptide. Peptidylamidoglycolate lyase is the PAM domain responsible for the Zn(II)-dependent dealkylation of the α-hydroxyglycine-containing precursor to the final α-amidated peptide. We report herein that cinnamic acid and cinnamic acid analogs are inhibitors or inactivators of PHM. The inactivation chemistry exhibited by the cinnamates exhibits all the attributes of a suicide-substrate. However, we find no evidence for the formation of an irreversible linkage between cinnamate and PHM in the inactivated enzyme. Our data support the reversible formation of a Michael adduct between an active site nucleophile and cinnamate that leads to inactive enzyme. Our data are of significance given that cinnamates are found in foods, perfumes, cosmetics and pharmaceuticals.

  5. Structural Basis for the Inactivation of Thermus thermophilus Proline Dehydrogenase by N-Propargylglycine†‡

    PubMed Central

    White, Tommi A.; Johnson, William H.; Whitman, Christian P.; Tanner, John J.

    2009-01-01

    The flavoenzyme proline dehydrogenase catalyzes the first step of proline catabolism, the oxidation of proline to pyrroline-5-carboxylate. Here we report the first crystal structure of an irreversibly inactivated proline dehydrogenase. The 1.9 Å resolution structure of Thermus thermophilus proline dehydrogenase inactivated by the mechanism-based inhibitor N-propargylglycine shows that N5 of the flavin cofactor is covalently connected to the ε-amino group of Lys99 via a 3-carbon linkage, consistent with the mass spectral analysis of the inactivated enzyme. The isoalloxazine ring has a butterfly angle of 25°, which suggests that the flavin cofactor is reduced. Two mechanisms can account for these observations. In both, N-propargylglycine is oxidized to N-propargyliminoglycine. In one mechanism, this α,β-unsaturated iminium compound is attacked by the N5 atom of the now reduced flavin to produce a 1,4-addition product. Schiff base formation between Lys99 and the imine of the 1,4-addition product releases glycine and links the enzyme to the modified flavin. In the second mechanism, hydrolysis of N-propargyliminoglycine yields propynal and glycine. A 1,4-addition reaction with propynal coupled with Schiff base formation between Lys99 and the carbonyl group tethers the enzyme to the flavin via a 3-carbon chain. The presumed non-enzymatic hydrolysis of N-propargyliminoglycine and the subsequent rebinding of propynal to the enzyme make the latter mechanism less likely. PMID:18426222

  6. Structural Basis for Reversible and Irreversible Inhibition of Human Cathepsin L by their Respective dipeptidyl glyoxal and diazomethylketone Inhibitors

    SciTech Connect

    R Shenoy; J Sivaraman

    2011-12-31

    Cathepsin L plays a key role in many pathophysiological conditions including rheumatoid arthritis, tumor invasion and metastasis, bone resorption and remodeling. Here we report the crystal structures of two analogous dipeptidyl inhibitor complexes which inhibit human cathepsin L in reversible and irreversible modes, respectively. To-date, there are no crystal structure reports of complexes of proteases with their glyoxal inhibitors or complexes of cathepsin L and their diazomethylketone inhibitors. These two inhibitors - inhibitor 1, an {alpha}-keto-{beta}-aldehyde and inhibitor 2, a diazomethylketone, have different groups in the S1 subsite. Inhibitor 1 [Z-Phe-Tyr (OBut)-COCHO], with a Ki of 0.6 nM, is the most potent, reversible, synthetic peptidyl inhibitor of cathepsin L reported to-date. The structure of the inhibitor 1 complex was refined up to 2.2 {angstrom} resolution. The structure of the complex of the inhibitor 2 [Z-Phe-Tyr (t-Bu)-diazomethylketone], an irreversible inhibitor that can inactivate cathepsin L at {micro}M concentrations, was refined up to 1.76 {angstrom} resolution. These two inhibitors have substrate-like interactions with the active site cysteine (Cys25). Inhibitor 1 forms a tetrahedral hemithioacetal adduct, whereas the inhibitor 2 forms a thioester with Cys25. The inhibitor 1 {beta}-aldehyde group is shown to make a hydrogen bond with catalytic His163, whereas the ketone carbonyl oxygen of the inhibitor 2 interacts with the oxyanion hole. tert-Butyl groups of both inhibitors are found to make several non-polar contacts with S' subsite residues of cathepsin L. These studies, combined with other complex structures of cathepsin L, reveal the structural basis for their potency and selectivity.

  7. Overcoming inactivation of the lung surfactant by serum proteins: a potential role for fluorocarbons?

    PubMed

    Krafft, Marie Pierre

    2015-08-14

    In many pulmonary conditions serum proteins interfere with the normal adsorption of components of the lung surfactant to the surface of the alveoli, resulting in lung surfactant inactivation, with potentially serious untoward consequences. Here, we review the strategies that have recently been designed in order to counteract the biophysical mechanisms of inactivation of the surfactant. One approach includes protein analogues or peptides that mimic the native proteins responsible for innate resistance to inactivation. Another perspective uses water-soluble additives, such as electrolytes and hydrophilic polymers that are prone to enhance adsorption of phospholipids. An alternative, more recent approach consists of using fluorocarbons, that is, highly hydrophobic inert compounds that were investigated for partial liquid ventilation, that modify interfacial properties and can act as carriers of exogenous lung surfactant. The latter approach that allows fluidisation of phospholipid monolayers while maintaining capacity to reach near-zero surface tension definitely warrants further investigation.

  8. N- vs. C-Domain Selectivity of Catalytic Inactivation of Human Angiotensin Converting Enzyme by Lisinopril-Coupled Transition Metal Chelates

    PubMed Central

    Hocharoen, Lalintip; Joyner, Jeff C.; Cowan, J. A.

    2014-01-01

    The N- and C-terminal domains of human somatic Angiotensin I Converting Enzyme (sACE-1) demonstrate distinct physiological functions, with resulting interest in the development of domain-selective inhibitors for specific therapeutic applications. Herein, the activity of lisinopril-coupled transition metal chelates were tested for both reversible binding and irreversible catalytic inactivation of sACE-1. C/N domain binding selectivity ratios ranged from 1 to 350, while rates of irreversible catalytic inactivation of the N- and C-domains were found to be significantly greater for the N-domain, suggesting a more optimal orientation of the M-chelate-lisinopril complexes within the active site of the N-domain of sACE-1. Finally, the combined effect of binding selectivity and inactivation selectivity was assessed for each catalyst (double-filter selectivity factors), and several catalysts were found to cause domain-selective catalytic inactivation. The results of this study demonstrate the ability to optimize the target selectivity of catalytic metallopeptides through both binding and orientation factors (double-filter effect). PMID:24228790

  9. Removal of sodium inactivation and block of sodium channels by chloramine-T in crayfish and squid giant axons.

    PubMed Central

    Huang, J. M.; Tanguy, J.; Yeh, J. Z.

    1987-01-01

    Modification of sodium channels by chloramine-T was examined in voltage clamped internally perfused crayfish and squid giant axons using the double sucrose gap and axial wire technique, respectively. Freshly prepared chloramine-T solution exerted two major actions on sodium channels: (a) an irreversible removal of the fast Na inactivation, and (b) a reversible block of the Na current. Both effects were observed when chloramine-T was applied internally or externally (5-10 mM) to axons. The first effect was studied in crayfish axons. We found that the removal of the fast Na inactivation did not depend on the states of the channel since the channel could be modified by chloramine-T at holding potential (from -80 to -100 mV) or at depolarized potential of -30 mV. After removal of fast Na inactivation, the slow inactivation mechanism was still present, and more channels could undergo slow inactivation. This result indicates that in crayfish axons the transition through the fast inactivated state is not a prerequisite for the slow inactivation to occur. During chloramine-T treatment, a distinct blocking phase occurred, which recovered upon washing out the drug. This second effect of chloramine-T was studied in detail in squid axons. After 24 h, chloramine-T solution lost its ability to remove fast inactivation but retained its blocking action. After removal of the fast Na inactivation, both fresh and aged chloramine-T solutions blocked the Na currents with a similar potency and in a voltage-dependent manner, being more pronounced at lower depolarizing potentials. A similar voltage-dependent block was observed with aged chloramine-T solution in an axon with intact inactivation. In contrast to the action of the fresh solution, the aged chloramine-T solution was found to accelerate the decay of Na currents.These results suggest that chloramine-T solution contains at least two active molecular forms that act at different sites in the Na channel. PMID:2444276

  10. Inactivation of Coprinus cinereus peroxidase by 4-chloroaniline during turnover: comparison with horseradish peroxidase and bovine lactoperoxidase.

    PubMed

    Chang, H C; Holland, R D; Bumpus, J A; Churchwell, M I; Doerge, D R

    1999-12-15

    The peroxidase from Coprinus cinereus (CPX) catalyzed oxidative oligomerization of 4-chloroaniline (4-CA) forming several products: N-(4-chlorophenyl)-benzoquinone monoamine (dimer D), 4,4'-dichloroazobenzene (dimer E); 2-(4-chloroanilino)-N-(4-chlorophenyl)-benzoquinone (trimer F); 2-amino-5-chlorobenzoquinone-di-4-chloroanil (trimer G); 2-(4-chloroanilino)-5-hydroxybenzoquinone-di-4-chloroanil (tetramer H) and 2-amino-5-(-4-chlroanilino)-benzoquinone-di-4-chloroanil (tetramer 1). In the presence of 4-CA and H2O2, CPX was irreversibly inactivated within 10 min. Inactivation of CPX in the presence of H2O2 was a time-dependent, first-order process when the concentration of 4-CA was varied between 0 and 2.5 mM. The apparent dissociation constant (Ki) for CPX and 4-CA was 0.71 mM. The pseudo-first order rate constant for inactivation (k(inact)), was 1.15 x 10(-2) s(-1). Covalent incorporation of 20 mole 14C-4-CA per mole of inactivated CPX was observed. The partition ratio was about 2200 when either 4-CA or H2O2 was used as the limiting substrate. These results show that 4-CA is a metabolically activated inactivator (i.e. a suicide substrate). Unmodified heme and hydroxymethyl heme were isolated from native, 4-CA-inactivated and H2O2-incubated CPX. Inactivation resulted in significant losses in both heme contents. Analysis of tryptic peptides from 4-CA-inactivated CPX by MALDI-TOF/ MS and UV-VIS spectrophotometry suggested that trimer G and tetramer H were the major 4-CA derivatives that were covalently bound, including to a peptide (MGDAGF-SPDEVVDLLAAHSLASQEGLNSAIFR) containing the heme binding site. These studies show that heme destruction and covalent modification of the polypeptide chain are both important for the inactivation of CPX. These results were compared with similar studies on 4-CA-inactivated horseradish peroxidase (HRP) and bovine lactoperoxidase (LPO) during the oxidation of 4-CA.

  11. Non-thermal irreversible electroporation for deep intracranial disorders.

    PubMed

    Garcia, Paulo A; Neal, Robert E; Rossmeisl, John H; Davalos, Rafael V

    2010-01-01

    Non-thermal irreversible electroporation (N-TIRE) is a new minimally invasive technique to kill undesirable tissue. We build on our previous intracranial studies in order to evaluate the possibility of using N-TIRE for deep intracranial disorders. In this manuscript we describe a minimally invasive computed tomography (CT) guided N-TIRE procedure in white matter. In addition, we report the electric field threshold needed for white matter ablation (630 - 875 V/cm) using four sets of twenty 50 µs pulses at a voltage-to-distance ratio of 1000 V/cm. We also confirm the non-thermal aspect of the technique with real time temperature data measured at the electrode-tissue interface. PMID:21095962

  12. Rat liver regeneration following ablation with irreversible electroporation

    PubMed Central

    Bruinsma, Bote G.; Jaramillo, Maria; Yarmush, Martin L.

    2016-01-01

    During the past decade, irreversible electroporation (IRE) ablation has emerged as a promising tool for the treatment of multiple diseases including hepatic cancer. However, the mechanisms behind the tissue regeneration following IRE ablation have not been investigated. Our results indicate that IRE treatment immediately kills the cells at the treatment site preserving the extracellular architecture, in effect causing in vivo decellularization. Over the course of 4 weeks, progenitor cell differentiation, through YAP and notch pathways, together with hepatocyte expansion led to almost complete regeneration of the ablated liver leading to the formation of hepatocyte like cells at the ablated zone. We did not observe significant scarring or tumor formation at the regenerated areas 6 months post IRE. Our study suggests a new model to study the regeneration of liver when the naïve extracellular matrix is decellularized in vivo with completely preserved extracellular architecture. PMID:26819842

  13. Immersion disinfection of irreversible hydrocolloid impressions. Part 1: Microbiology.

    PubMed

    Schwartz, R S; Bradley, D V; Hilton, T J; Kruse, S K

    1994-01-01

    This study evaluated the effectiveness of four disinfectants for irreversible hydrocolloid impressions. Impressions were made of a sterile metal model of the maxillary arch that had been contaminated with one of the following bacteria: Staphylococcus aureus, Salmonella choleraesuis, Pseudomonas aeruginosa, Mycobacterium bovis, or Bacillus subtilis. The impressions were cultured before and after immersion in one of the following disinfectants: lodofive, OMC II, 0.525% sodium hypochlorite, or Alcide LD. Alcide LD achieved a 4-log10 (99.99%) or greater reduction in colony forming units for all five organisms plus mixed oral flora. Sodium hypochlorite achieved a 4-log10 reduction in three of the five organisms and mixed oral flora. Iodofive and OMC II were ineffective against all test organisms and mixed oral flora. PMID:7802909

  14. Immersion disinfection of irreversible hydrocolloid impressions. Part 1: Microbiology.

    PubMed

    Schwartz, R S; Bradley, D V; Hilton, T J; Kruse, S K

    1994-01-01

    This study evaluated the effectiveness of four disinfectants for irreversible hydrocolloid impressions. Impressions were made of a sterile metal model of the maxillary arch that had been contaminated with one of the following bacteria: Staphylococcus aureus, Salmonella choleraesuis, Pseudomonas aeruginosa, Mycobacterium bovis, or Bacillus subtilis. The impressions were cultured before and after immersion in one of the following disinfectants: lodofive, OMC II, 0.525% sodium hypochlorite, or Alcide LD. Alcide LD achieved a 4-log10 (99.99%) or greater reduction in colony forming units for all five organisms plus mixed oral flora. Sodium hypochlorite achieved a 4-log10 reduction in three of the five organisms and mixed oral flora. Iodofive and OMC II were ineffective against all test organisms and mixed oral flora.

  15. Ac irreversibility line of bismuth-based high temperature superconductors

    SciTech Connect

    Mehdaoui, A.; Beille, J.; Berling, D.; Loegel, B.; Noudem, J.G.; Tournier, R.

    1997-09-01

    We discuss the magnetic properties of lead doped Bi-2223 bulk samples obtained through combined magnetic melt texturing and hot pressing (MMTHP). The ac complex susceptibility measurements are achieved over a broad ac field range (1 Oe{lt}h{sub ac}{lt}100 Oe) and show highly anisotropic properties. The intergranular coupling is improved in the direction perpendicular to the applied stress and magnetic field direction, and an intragranular loss peak is observed for the first time. A comparison is made with other bismuth-based compounds and it is shown that the MMTHP process shifts the ac irreversibility line (ac IL) toward higher fields. It is also shown that all the ac IL{close_quote}s for quasi 2D bismuth-based compounds show a nearly quadratic temperature dependence and deviate therefore strongly from the linear behavior observed in quasi 3D compounds and expected from a critical state model.{copyright} {ital 1997 Materials Research Society.}

  16. Sub-kBT micro-electromechanical irreversible logic gate

    PubMed Central

    López-Suárez, M.; Neri, I.

    2016-01-01

    In modern computers, computation is performed by assembling together sets of logic gates. Popular gates like AND, OR and XOR, processing two logic inputs and yielding one logic output, are often addressed as irreversible logic gates, where the sole knowledge of the output logic value is not sufficient to infer the logic value of the two inputs. Such gates are usually believed to be bounded to dissipate a finite minimum amount of energy determined by the input–output information difference. Here we show that this is not necessarily the case, by presenting an experiment where a OR logic gate, realized with a micro-electromechanical cantilever, is operated with energy well below the expected limit, provided the operation is slow enough and frictional phenomena are properly addressed. PMID:27350333

  17. Irreversible electroporation for the treatment of pancreatic neuroendocrine tumors

    PubMed Central

    Papamichail, Michail; Ali, Amir; Pizanias, Michail; Peddu, Praveen; Karani, John

    2016-01-01

    Backgrounds/Aims Resection or enucleation is currently the treatment of choice for small pancreatic neuroendocrine tumors (NETs). Irreversible electroporation is a novel ablative method that is used for locally advanced pancreatic adenocarcinoma, but little data exists for its use for pancreatic NETs. We report an early experience of IRE for early pancreatic NETs. Methods Between April 2014 and March 2015, 3 patients with small (<2 cm) pancreatic NETs were treated with percutaneous IRE. Results There were no adverse effects during the procedure. Mean hospital stay was 2.6 days. All patients remained disease free on 12-19 months follow up. One patient developed recurrent pancreatitis with pseudocyst formation. Conclusions IRE for small tumors of the pancreas is practical and may offer advantages over other thermal ablative techniques, since it preserves vital structures such as blood vessels, bile and pancreatic ducts. Further data regarding the long term disease free interval is required to establish efficacy.

  18. Sub-kBT micro-electromechanical irreversible logic gate.

    PubMed

    López-Suárez, M; Neri, I; Gammaitoni, L

    2016-01-01

    In modern computers, computation is performed by assembling together sets of logic gates. Popular gates like AND, OR and XOR, processing two logic inputs and yielding one logic output, are often addressed as irreversible logic gates, where the sole knowledge of the output logic value is not sufficient to infer the logic value of the two inputs. Such gates are usually believed to be bounded to dissipate a finite minimum amount of energy determined by the input-output information difference. Here we show that this is not necessarily the case, by presenting an experiment where a OR logic gate, realized with a micro-electromechanical cantilever, is operated with energy well below the expected limit, provided the operation is slow enough and frictional phenomena are properly addressed. PMID:27350333

  19. Typical pure nonequilibrium steady states and irreversibility for quantum transport.

    PubMed

    Monnai, Takaaki; Yuasa, Kazuya

    2016-07-01

    It is known that each single typical pure state in an energy shell of a large isolated quantum system well represents a thermal equilibrium state of the system. We show that such typicality holds also for nonequilibrium steady states (NESS's). We consider a small quantum system coupled to multiple infinite reservoirs. In the long run, the total system reaches a unique NESS. We identify a large Hilbert space from which pure states of the system are to be sampled randomly and show that the typical pure states well describe the NESS. We also point out that the irreversible relaxation to the unique NESS is important to the typicality of the pure NESS's. PMID:27575115

  20. Irreversible electroporation for the treatment of pancreatic neuroendocrine tumors

    PubMed Central

    Papamichail, Michail; Ali, Amir; Pizanias, Michail; Peddu, Praveen; Karani, John

    2016-01-01

    Backgrounds/Aims Resection or enucleation is currently the treatment of choice for small pancreatic neuroendocrine tumors (NETs). Irreversible electroporation is a novel ablative method that is used for locally advanced pancreatic adenocarcinoma, but little data exists for its use for pancreatic NETs. We report an early experience of IRE for early pancreatic NETs. Methods Between April 2014 and March 2015, 3 patients with small (<2 cm) pancreatic NETs were treated with percutaneous IRE. Results There were no adverse effects during the procedure. Mean hospital stay was 2.6 days. All patients remained disease free on 12-19 months follow up. One patient developed recurrent pancreatitis with pseudocyst formation. Conclusions IRE for small tumors of the pancreas is practical and may offer advantages over other thermal ablative techniques, since it preserves vital structures such as blood vessels, bile and pancreatic ducts. Further data regarding the long term disease free interval is required to establish efficacy. PMID:27621748

  1. Focal Therapy of Prostate Cancer Using Irreversible Electroporation.

    PubMed

    Valerio, Massimo; Ahmed, Hashim U; Emberton, Mark

    2015-09-01

    Focal therapy is a novel strategy that attempts to enhance the therapeutic ratio of standard radical treatment in prostate cancer. Irreversible electroporation (IRE) has some inherent characteristics that may be ideal for focal therapy. Precise confined ablation in the treatment area obtained via nonthermal damage with potential for minimal toxicity to surrounding structures may lead to optimal treatment with improved preservation of continence and erectile function. Initial data of focal IRE of the prostate are encouraging although further assessment is awaited to confirm these findings using robust methodology. In this article, we provide a comprehensive step-by-step description of our technique to deliver focal IRE in selected men with localized prostate cancer located in a discrete area of the prostate.

  2. Sub-kBT micro-electromechanical irreversible logic gate

    NASA Astrophysics Data System (ADS)

    López-Suárez, M.; Neri, I.; Gammaitoni, L.

    2016-06-01

    In modern computers, computation is performed by assembling together sets of logic gates. Popular gates like AND, OR and XOR, processing two logic inputs and yielding one logic output, are often addressed as irreversible logic gates, where the sole knowledge of the output logic value is not sufficient to infer the logic value of the two inputs. Such gates are usually believed to be bounded to dissipate a finite minimum amount of energy determined by the input-output information difference. Here we show that this is not necessarily the case, by presenting an experiment where a OR logic gate, realized with a micro-electromechanical cantilever, is operated with energy well below the expected limit, provided the operation is slow enough and frictional phenomena are properly addressed.

  3. Consequences of Irreversibility in Fundamental Models of Transcription

    NASA Astrophysics Data System (ADS)

    Sevier, Stuart; Levine, Herbert

    2015-03-01

    The ability to watch biochemical events play out at the single-molecule level has led to the discovery that transcription occurs in a noisy, ``bursty'' manner. Recently, as the single-molecule lens is placed over a larger number of organisms and genes, relationships between mean expression and noise beyond the ``bursty'' paradigm have emerged. Through a master-equation formulation of transcription we have found that many powerful physical principles relating to irreversibility seem to play a central role in the newly uncovered trends. Specifically, the relationships between mean expression and noise appears to be a direct consequence of network currents. We discuss how emphasizing the underlying principles in the models can explain recent experimental data and lead to a generalized view of transcription.

  4. Irreversible degradation of quantum coherence under relativistic motion

    NASA Astrophysics Data System (ADS)

    Wang, Jieci; Tian, Zehua; Jing, Jiliang; Fan, Heng

    2016-06-01

    We study the dynamics of quantum coherence under Unruh thermal noise and seek under which condition the coherence can be frozen in a relativistic setting. We find that the frozen condition is either (i) the initial state is prepared as an incoherence state or (ii) the detectors have no interaction with the external field. That is to say, the decoherence of the detectors' quantum state is irreversible under the influence of thermal noise induced by Unruh radiation. It is shown that quantum coherence approaches zero only in the limit of an infinite acceleration, while quantum entanglement could reduce to zero for a finite acceleration. It is also demonstrated that the robustness of quantum coherence is better than entanglement under the influence of the atom-field interaction for an extremely large acceleration. Therefore, quantum coherence is more robust than entanglement in an accelerating system and the coherence-type quantum resources are more accessible for relativistic quantum information processing tasks.

  5. Advertising and Irreversible Opinion Spreading in Complex Social Networks

    NASA Astrophysics Data System (ADS)

    Candia, Julián

    Irreversible opinion spreading phenomena are studied on small-world and scale-free networks by means of the magnetic Eden model, a nonequilibrium kinetic model for the growth of binary mixtures in contact with a thermal bath. In this model, the opinion of an individual is affected by those of their acquaintances, but opinion changes (analogous to spin flips in an Ising-like model) are not allowed. We focus on the influence of advertising, which is represented by external magnetic fields. The interplay and competition between temperature and fields lead to order-disorder transitions, which are found to also depend on the link density and the topology of the complex network substrate. The effects of advertising campaigns with variable duration, as well as the best cost-effective strategies to achieve consensus within different scenarios, are also discussed.

  6. Large-cell Monte Carlo renormalization of irreversible growth processes

    NASA Technical Reports Server (NTRS)

    Nakanishi, H.; Family, F.

    1985-01-01

    Monte Carlo sampling is applied to a recently formulated direct-cell renormalization method for irreversible, disorderly growth processes. Large-cell Monte Carlo renormalization is carried out for various nonequilibrium problems based on the formulation dealing with relative probabilities. Specifically, the method is demonstrated by application to the 'true' self-avoiding walk and the Eden model of growing animals for d = 2, 3, and 4 and to the invasion percolation problem for d = 2 and 3. The results are asymptotically in agreement with expectations; however, unexpected complications arise, suggesting the possibility of crossovers, and in any case, demonstrating the danger of using small cells alone, because of the very slow convergence as the cell size b is extrapolated to infinity. The difficulty of applying the present method to the diffusion-limited-aggregation model, is commented on.

  7. A systematic comprehensive approach to management of irreversible facial paralysis.

    PubMed

    Douglas, Raymond S; Gausas, Roberta E

    2003-02-01

    Irreversible facial palsy (IFP) presents a multitude of problems arising from a paretic periorbital and facial complex, the solutions to which cross the spectrum of multiple specialties. The process of facial rehabilitation can be simplified by subdividing the face into functional units. These units consist of the brow complex, the periorbital complex, the midface complex, and the lower face/oral complex. Although all of these units are interrelated and influence each other, careful study of the deformity and symptoms of each unit yields a coherent approach and customized surgical plan. The following provides a complete evaluation method for the surgeon to review and customize an approach to the individual patient's needs and desires. Facial rehabilitation must be tailored to each individual, addressing both functional as well as aesthetic concerns for each facial unit.

  8. The Social Cost of Stochastic and Irreversible Climate Change

    NASA Astrophysics Data System (ADS)

    Cai, Y.; Judd, K. L.; Lontzek, T.

    2013-12-01

    Many scientists are worried about climate change triggering abrupt and irreversible events leading to significant and long-lasting damages. For example, a rapid release of methane from permafrost may lead to amplified global warming, and global warming may increase the frequency and severity of heavy rainfall or typhoon, destroying large cities and killing numerous people. Some elements of the climate system which might exhibit such a triggering effect are called tipping elements. There is great uncertainty about the impact of anthropogenic carbon and tipping elements on future economic wellbeing. Any rational policy choice must consider the great uncertainty about the magnitude and timing of global warming's impact on economic productivity. While the likelihood of tipping points may be a function of contemporaneous temperature, their effects are long lasting and might be independent of future temperatures. It is assumed that some of these tipping points might occur even in this century, but also that their duration and post-tipping impact are uncertain. A faithful representation of the possibility of tipping points for the calculation of social cost of carbon would require a fully stochastic formulation of irreversibility, and accounting for the deep layer of uncertainties regarding the duration of the tipping process and also its economic impact. We use DSICE, a DSGE extension of the DICE2007 model of William Nordhaus, which incorporates beliefs about the uncertain economic impact of possible climate tipping events and uses empirically plausible parameterizations of Epstein-Zin preferences to represent attitudes towards risk. We find that the uncertainty associated with anthropogenic climate change imply carbon taxes much higher than implied by deterministic models. This analysis indicates that the absence of uncertainty in DICE2007 and similar IAM models may result in substantial understatement of the potential benefits of policies to reduce GHG emissions.

  9. Two studies of nonlinear processes in irreversible thermodynamics

    SciTech Connect

    Kestin, J.

    1992-01-01

    The investigation dealt with two lines of research into two well-defined problems of engineering science: a study of two-phase flow and of nonelastic deformations in structural solids. Both topics fall into the broad field of nonlinear irreversible processes. The study of two-phase flow resulted in a complete topological analysis of the canonical mathematical model of one-dimensional flow of a mixture of two phases, now predominantly used in industry, especially the nuclear industry. The topological analysis is confronted with the practice of discretizing the analytic model for the purpose of formulating a numerical computer code. It is shown that in the presence of singular points in the phase space of the differential equations of the model there occur spurious numerical solutions. Such solutions may contain segments which are correct approximations to the exact trajectory, but always include branches which are totally false and misleading. The topological method allows the operator to formulate a subroutine which eliminates spurious solutions. The study of inelastic deformations, i.e., of irreversible processes in structural solids provided a consistent presentation of the local-state approximation erroneously called the method of local equilibrium in the literature. The key concept which impedes the correct use of thermodynamics in this field is the definition of a measurable entropy of a nonequilibrium state. The local state approximation solves this problem by assigning to a nonequilibrium state n the entropy of the accompanying equilibrium state e. The two states, n and e, are linked by having the same values of a specified set of extensive properties. The same principle settles the comparison between the intensive properties of n and e, which are different.

  10. Inactivation of ribulosebisphosphate carboxylase/oxygenase from Rhodospirillum rubrum and spinach with the new affinity label 2-bromo-1,5-dihydroxy-3-pentanone 1,5-bisphosphate

    SciTech Connect

    Donnelly, M.I.; Hartman, F.C.

    1981-11-16

    In an attempt to identify the active-site base believed to initiate catalysis by ribulosebisphosphate carboxylase, we have synthesized 2-bromo-1, 5-dihydroxy-3-pentanone 1,5-bisphosphate, a reactive analogue of a postulated intermediate of carboxylation. Although highly unstable, this compound can be shown to inactivate the carboxylases from both Rhodospirillum rubrum and spinach rapidly and irreversibly. Inactivation follows pseudo first-order kinetics, shows rate saturation and is greatly reduced by saturating amounts of the competitive inhibitor, 2-carboxyribitol 1,5-bisphosphate. The incorporation of reagent, quantified by reducing the modified carboxylases with (/sup 3/H)NaBH/sub 4/, shows that inactivation results from the modification of approximately one residue per catalytic subunit of the Rhodospirillum rubrum enzyme and less than one residue per protomeric unit of the spinach enzyme.

  11. GnRH Neuron-Specific Ablation of Gαq/11 Results in Only Partial Inactivation of the Neuroendocrine-Reproductive Axis in Both Male and Female Mice: In Vivo Evidence for Kiss1r-Coupled Gαq/11-Independent GnRH Secretion

    PubMed Central

    Navarro, Víctor M.; Ahow, Maryse; Pampillo, Macarena; Nash, Connor; Fayazi, Mehri; Calder, Michele; Elbert, Adrienne; Urbanski, Henryk F.; Wettschureck, Nina; Offermanns, Stefan; Carroll, Rona S.; Bhattacharya, Moshmi; Tobet, Stuart A.; Kaiser, Ursula B.

    2015-01-01

    The gonadotropin-releasing hormone (GnRH) is the master regulator of fertility and kisspeptin (KP) is a potent trigger of GnRH secretion from GnRH neurons. KP signals via KISS1R, a Gαq/11-coupled receptor, and mice bearing a global deletion of Kiss1r (Kiss1r−/−) or a GnRH neuron-specific deletion of Kiss1r (Kiss1rd/d) display hypogonadotropic hypogonadism and infertility. KISS1R also signals via β-arrestin, and in mice lacking β-arrestin-1 or -2, KP-triggered GnRH secretion is significantly diminished. Based on these findings, we hypothesized that ablation of Gαq/11 in GnRH neurons would diminish but not completely block KP-triggered GnRH secretion and that Gαq/11-independent GnRH secretion would be sufficient to maintain fertility. To test this, Gnaq (encodes Gαq) was selectively inactivated in the GnRH neurons of global Gna11 (encodes Gα11)-null mice by crossing Gnrh-Cre and Gnaqfl/fl;Gna11−/− mice. Experimental Gnaqfl/fl;Gna11−/−;Gnrh-Cre (Gnaqd/d) and control Gnaqfl/fl;Gna11−/− (Gnaqfl/fl) littermate mice were generated and subjected to reproductive profiling. This process revealed that testicular development and spermatogenesis, preputial separation, and anogenital distance in males and day of vaginal opening and of first estrus in females were significantly less affected in Gnaqd/d mice than in previously characterized Kiss1r−/− or Kiss1rd/d mice. Additionally, Gnaqd/d males were subfertile, and although Gnaqd/d females did not ovulate spontaneously, they responded efficiently to a single dose of gonadotropins. Finally, KP stimulation triggered a significant increase in gonadotropins and testosterone levels in Gnaqd/d mice. We therefore conclude that the milder reproductive phenotypes and maintained responsiveness to KP and gonadotropins reflect Gαq/11-independent GnRH secretion and activation of the neuroendocrine-reproductive axis in Gnaqd/d mice. SIGNIFICANCE STATEMENT The gonadotropin-releasing hormone (GnRH) is the master

  12. Second Generation Inactivated Eastern Equine Encephalitis Virus Vaccine Candidates Protect Mice against a Lethal Aerosol Challenge

    PubMed Central

    Honnold, Shelley P.; Bakken, Russell R.; Fisher, Diana; Lind, Cathleen M.; Cohen, Jeffrey W.; Eccleston, Lori T.; Spurgers, Kevin B.; Maheshwari, Radha K.; Glass, Pamela J.

    2014-01-01

    Currently, there are no FDA-licensed vaccines or therapeutics for eastern equine encephalitis virus (EEEV) for human use. We recently developed several methods to inactivate CVEV1219, a chimeric live-attenuated eastern equine encephalitis virus (EEEV). Dosage and schedule studies were conducted to evaluate the immunogenicity and protective efficacy of three potential second-generation inactivated EEEV (iEEEV) vaccine candidates in mice: formalin-inactivated CVEV1219 (fCVEV1219), INA-inactivated CVEV1219 (iCVEV1219) and gamma-irradiated CVEV1219 (gCVEV1219). Both fCVEV1219 and gCVEV1219 provided partial to complete protection against an aerosol challenge when administered by different routes and schedules at various doses, while iCVEV1219 was unable to provide substantial protection against an aerosol challenge by any route, dose, or schedule tested. When evaluating antibody responses, neutralizing antibody, not virus specific IgG or IgA, was the best correlate of protection. The results of these studies suggest that both fCVEV1219 and gCVEV1219 should be evaluated further and considered for advancement as potential second-generation inactivated vaccine candidates for EEEV. PMID:25116127

  13. Identifying and Inactivating Bacterial Spores

    NASA Technical Reports Server (NTRS)

    Newcombe, David; Dekas, Anne; Venkateswaran, Kasthuri

    2009-01-01

    Problems associated with, and new strategies for, inactivating resistant organisms like Bacillus canaveralius (found at Kennedy Space Center during a survey of three NASA cleanrooms) have been defined. Identifying the particular component of the spore that allows its heightened resistance can guide the development of sterilization procedures that are targeted to the specific molecules responsible for resistance, while avoiding using unduly harsh methods that jeopardize equipment. The key element of spore resistance is a multilayered protein shell that encases the spore called the spore coat. The coat of the best-studied spore-forming microbe, B. subtilis, consists of at least 45 proteins, most of which are poorly characterized. Several protective roles for the coat are well characterized including resistance to desiccation, large toxic molecules, ortho-phthalaldehyde, and ultraviolet (UV) radiation. One important long-term specific goal is an improved sterilization procedure that will enable NASA to meet planetary protection requirements without a terminal heat sterilization step. This would support the implementation of planetary protection policies for life-detection missions. Typically, hospitals and government agencies use biological indicators to ensure the quality control of sterilization processes. The spores of B. canaveralius that are more resistant to osmotic stress would serve as a better biological indicator for potential survival than those in use currently.

  14. Ribosome Inactivating Proteins from Rosaceae.

    PubMed

    Shang, Chenjing; Rougé, Pierre; Van Damme, Els J M

    2016-01-01

    Ribosome-inactivating proteins (RIPs) are widespread among higher plants of different taxonomic orders. In this study, we report on the RIP sequences found in the genome/transcriptome of several important Rosaceae species, including many economically important edible fruits such as apple, pear, peach, apricot, and strawberry. All RIP domains from Rosaceae share high sequence similarity with conserved residues in the catalytic site and the carbohydrate binding sites. The genomes of Malus domestica and Pyrus communis contain both type 1 and type 2 RIP sequences, whereas for Prunus mume, Prunus persica, Pyrus bretschneideri, and Pyrus communis a complex set of type 1 RIP sequences was retrieved. Heterologous expression and purification of the type 1 as well as the type 2 RIP from apple allowed to characterize the biological activity of the proteins. Both RIPs from Malus domestica can inhibit protein synthesis. Furthermore, molecular modelling suggests that RIPs from Rosaceae possess three-dimensional structures that are highly similar to the model proteins and can bind to RIP substrates. Screening of the recombinant type 2 RIP from apple on a glycan array revealed that this type 2 RIP interacts with terminal sialic acid residues. Our data suggest that the RIPs from Rosaceae are biologically active proteins. PMID:27556443

  15. Population Dynamics of Viral Inactivation

    NASA Astrophysics Data System (ADS)

    Freeman, Krista; Li, Dong; Behrens, Manja; Streletzky, Kiril; Olsson, Ulf; Evilevitch, Alex

    We have investigated the population dynamics of viral inactivation in vitrousing time-resolved cryo electron microscopy combined with light and X-ray scattering techniques. Using bacteriophage λ as a model system for pressurized double-stranded DNA viruses, we found that virions incubated with their cell receptor eject their genome in a stochastic triggering process. The triggering of DNA ejection occurs in a non synchronized manner after the receptor addition, resulting in an exponential decay of the number of genome-filled viruses with time. We have explored the characteristic time constant of this triggering process at different temperatures, salt conditions, and packaged genome lengths. Furthermore, using the temperature dependence we determined an activation energy for DNA ejections. The dependences of the time constant and activation energy on internal DNA pressure, affected by salt conditions and encapsidated genome length, suggest that the triggering process is directly dependent on the conformational state of the encapsidated DNA. The results of this work provide insight into how the in vivo kinetics of the spread of viral infection are influenced by intra- and extra cellular environmental conditions. This material is based upon work supported by the National Science Foundation Graduate Research Fellowship under Grant No. DGE-1252522.

  16. Ribosome Inactivating Proteins from Rosaceae.

    PubMed

    Shang, Chenjing; Rougé, Pierre; Van Damme, Els J M

    2016-01-01

    Ribosome-inactivating proteins (RIPs) are widespread among higher plants of different taxonomic orders. In this study, we report on the RIP sequences found in the genome/transcriptome of several important Rosaceae species, including many economically important edible fruits such as apple, pear, peach, apricot, and strawberry. All RIP domains from Rosaceae share high sequence similarity with conserved residues in the catalytic site and the carbohydrate binding sites. The genomes of Malus domestica and Pyrus communis contain both type 1 and type 2 RIP sequences, whereas for Prunus mume, Prunus persica, Pyrus bretschneideri, and Pyrus communis a complex set of type 1 RIP sequences was retrieved. Heterologous expression and purification of the type 1 as well as the type 2 RIP from apple allowed to characterize the biological activity of the proteins. Both RIPs from Malus domestica can inhibit protein synthesis. Furthermore, molecular modelling suggests that RIPs from Rosaceae possess three-dimensional structures that are highly similar to the model proteins and can bind to RIP substrates. Screening of the recombinant type 2 RIP from apple on a glycan array revealed that this type 2 RIP interacts with terminal sialic acid residues. Our data suggest that the RIPs from Rosaceae are biologically active proteins.

  17. Susceptibility of the early Earth to irreversible glaciation caused by carbon dioxide clouds.

    PubMed

    Caldeira, K; Kasting, J F

    1992-09-17

    Simple energy-balance climate models of the Budyko/Sellers type predict that a small (2-5%) decrease in solar output could result in runaway glaciation on the Earth. But solar fluxes 25-30% lower early in the Earth's history apparently did not lead to this result. One currently favoured explanation is that high partial pressures of carbon dioxide, caused by higher volcanic outgassing rates and/or slower rates of silicate weathering, created a large enough greenhouse effect to keep the planet warm. This does not resolve the problem of climate stability, however, because as we argue here, the oceans can freeze much more quickly than CO2 can accumulate in the atmosphere. Had such a transient global glaciation occurred in the distant past when solar luminosity was low, it might have been irreversible because of the formation of highly reflective CO2 clouds, similar to those encountered in climate simulations of early Mars. Our simulations of the early Earth, incorporating the possible formation of such clouds, suggest that the Earth might not be habitable today had it not been warm during the first part of its history.

  18. Percutaneous Irreversible Electroporation Lung Ablation: Preliminary Results in a Porcine Model

    SciTech Connect

    Deodhar, Ajita; Monette, Sebastien; Single, Gordon W.; Hamilton, William C.; Thornton, Raymond H.; Sofocleous, Constantinos T.; Maybody, Majid; Solomon, Stephen B.

    2011-12-15

    Objective: Irreversible electroporation (IRE) uses direct electrical pulses to create permanent 'pores' in cell membranes to cause cell death. In contrast to conventional modalities, IRE has a nonthermal mechanism of action. Our objective was to study the histopathological and imaging features of IRE in normal swine lung. Materials and Methods: Eleven female swine were studied for hyperacute (8 h), acute (24 h), subacute (96 h), and chronic (3 week) effects of IRE ablation in lung. Paired unipolar IRE applicators were placed under computed tomography (CT) guidance. Some applicators were deliberately positioned near bronchovascular structures. IRE pulse delivery was synchronized with the cardiac rhythm only when ablation was performed within 2 cm of the heart. Contrast-enhanced CT scan was performed immediately before and after IRE and at 1 and 3 weeks after IRE ablation. Representative tissue was stained with hematoxylin and eosin for histopathology. Results: Twenty-five ablations were created: ten hyperacute, four acute, and three subacute ablations showed alveolar edema and necrosis with necrosis of bronchial, bronchiolar, and vascular epithelium. Bronchovascular architecture was maintained. Chronic ablations showed bronchiolitis obliterans and alveolar interstitial fibrosis. Immediate post-procedure CT images showed linear or patchy density along the applicator tract. At 1 week, there was consolidation that resolved partially or completely by 3 weeks. Pneumothorax requiring chest tube developed in two animals; no significant cardiac arrhythmias were noted. Conclusion: Our preliminary porcine study demonstrates the nonthermal and extracellular matrix sparing mechanism of action of IRE. IRE is a potential alternative to thermal ablative modalities.

  19. Histological and finite element analysis of cell death due to irreversible electroporation.

    PubMed

    Long, G; Bakos, G; Shires, P K; Gritter, L; Crissman, J W; Harris, J L; Clymer, J W

    2014-12-01

    Irreversible electroporation (IRE) has been shown to be an effective method of killing cells locally. In contrast to radiofrequency ablation, the mechanism by which cells are thought to die via IRE is the creation of pores in cell membranes, without substantial increase in tissue temperature. To determine the degree to which cell death is non-thermal, we evaluated IRE in porcine hepatocytes in vivo. Using pulse widths of 10 µs, bursts of 3 kV square-wave pulses were applied through a custom probe to the liver of an anesthetized pig. Affected tissue was evaluated histologically via stainings of hematoxylin & eosin (H&E), nitroblue tetrazolium (NBT) to monitor cell respiration and TUNEL to gauge apoptosis. Temperature was measured during the application of electroporation, and heat transfer was modeled via finite element analysis. Cell death was calculated via Arrhenius kinetics. Four distinct zones were observed within the ring return electrode; heat-fixed tissue, coagulation, necrotic, and viable. The Arrhenius damage integral estimated complete cell death only in the first zone, where the temperature exceeded 70°C, and partial or no cell death in the other zones, where maximum temperature was approximately 45°C. Except for a limited area near the electrode tip, cell death in IRE is predominantly due to a non-thermal mechanism.

  20. Irreversible binding of an anticancer compound (BI-94) to plasma proteins

    PubMed Central

    Gautam, Nagsen; Thakare, Rhishikesh; Rana, Sandeep; Natarajan, Amarnath; Alnouti, Yazen

    2015-01-01

    1. We investigated the mechanisms responsible for the in vivo instability of a benzofurazan compound BI-94 (NSC228148) with potent anti-cancer activity. 2. BI-94 was stable in MeOH, water, and in various buffers at pHs 2.5–5, regardless of the buffer composition. In contrast, BI-94 was unstable in NaOH and at pHs 7–9, regardless of the buffer composition. BI-94 disappeared immediately after spiking into mice, rat, monkey, and human plasma. BI-94 stability in plasma can be only partially restored by acidifying it, which indicated other mechanisms in addition to pH for BI-94 instability in plasma. 3. BI-94 formed adducts with the trapping agents, glutathione (GSH) and N-acetylcysteine (NAC), in vivo and in vitro via nucleophilic aromatic substitution reaction. The kinetics of adduct formation showed that neutral or physiological pHs enhanced and accelerated GSH and NAC adduct formation with BI-94, whereas acidic pHs prevented it. Therefore, physiological pHs not only altered BI-94 chemical stability but also enhanced adduct formation with endogenous nucleophiles. In addition, adduct formation with human serum albumin-peptide 3 (HSA-T3) at the Cys34 position was demonstrated. 4. In conclusion, BI-94 was unstable at physiological conditions due to chemical instability and irreversible binding to plasma proteins. PMID:25869245

  1. Irreversible and reversible reactive chromatography: analytical solutions and moment analysis for rectangular pulse injections.

    PubMed

    Bibi, Sameena; Qamar, Shamsul; Seidel-Morgenstern, Andreas

    2015-03-13

    This work is concerned with the analysis of models for linear reactive chromatography describing irreversible A→B and reversible A↔B reactions. In contrast to previously published results rectangular reactant pulses are injected into initially empty or pre-equilibrated columns assuming both Dirichlet and Danckwerts boundary conditions. The models consist of two partial differential equations, accounting for convection, longitudinal dispersion and first order chemical reactions. Due to the effect of involved mechanisms on solute transport, analytical and numerical solutions of the models could be helpful to understand, design and optimize chromatographic reactors. The Laplace transformation is applied to solve the model equations analytically for linear adsorption isotherms. Statistical temporal moments are derived from solutions in the Laplace domain. Analytical results are compared with numerical predictions generated using a high-resolution finite volume scheme for two sets of boundary conditions. Several case studies are carried out to analyze reactive liquid chromatographic processes for a wide range of mass transfer and reaction kinetics. Good agreements in the results validate the correctness of the analytical solutions and accuracy of the proposed numerical algorithm.

  2. Susceptibility of the early Earth to irreversible glaciation caused by carbon dioxide clouds.

    PubMed

    Caldeira, K; Kasting, J F

    1992-09-17

    Simple energy-balance climate models of the Budyko/Sellers type predict that a small (2-5%) decrease in solar output could result in runaway glaciation on the Earth. But solar fluxes 25-30% lower early in the Earth's history apparently did not lead to this result. One currently favoured explanation is that high partial pressures of carbon dioxide, caused by higher volcanic outgassing rates and/or slower rates of silicate weathering, created a large enough greenhouse effect to keep the planet warm. This does not resolve the problem of climate stability, however, because as we argue here, the oceans can freeze much more quickly than CO2 can accumulate in the atmosphere. Had such a transient global glaciation occurred in the distant past when solar luminosity was low, it might have been irreversible because of the formation of highly reflective CO2 clouds, similar to those encountered in climate simulations of early Mars. Our simulations of the early Earth, incorporating the possible formation of such clouds, suggest that the Earth might not be habitable today had it not been warm during the first part of its history. PMID:11540934

  3. Microbial Inactivation by Ultrasound Assisted Supercritical Fluids

    NASA Astrophysics Data System (ADS)

    Benedito, Jose; Ortuño, Carmen; Castillo-Zamudio, Rosa Isela; Mulet, Antonio

    A method combining supercritical carbon dioxide (SC-CO2) and high power ultrasound (HPU) has been developed and tested for microbial/enzyme inactivation purposes, at different process conditions for both liquid and solid matrices. In culture media, using only SC-CO2, the inactivation rate of E. coli and S. cerevisiae increased with pressure and temperature; and the total inactivation (7-8 log-cycles) was attained after 25 and 140 min of SC-CO2 (350 bar, 36 °C) treatment, respectively. Using SC-CO2+HPU, the time for the total inactivation of both microorganisms was reduced to only 1-2 min, at any condition selected. The SC-CO2+HPU inactivation of both microorganisms was slower in juices (avg. 4.9 min) than in culture media (avg. 1.5 min). In solid samples (chicken, turkey ham and dry-cured pork cured ham) treated with SC-CO2 and SC-CO2+HPU, the inactivation rate of E. coli increased with temperature. The application of HPU to the SC-CO2 treatments accelerated the inactivation rate of E. coli and that effect was more pronounced in treatments with isotonic solution surrounding the solid food samples. The application of HPU enhanced the SC-CO2 inactivation mechanisms of microorganisms, generating a vigorous agitation that facilitated the CO2 solubilization and the mass transfer process. The cavitation generated by HPU could damage the cell walls accelerating the extraction of vital constituents and the microbial death. Thus, using the combined technique, reasonable industrial processing times and mild process conditions could be used which could result into a cost reduction and lead to the minimization in the food nutritional and organoleptic changes.

  4. SCAM analysis reveals a discrete region of the pore turret that modulates slow inactivation in Kv1.5.

    PubMed

    Eduljee, Cyrus; Claydon, Thomas W; Viswanathan, Vijay; Fedida, David; Kehl, Steven J

    2007-03-01

    In Kv1.5, protonation of histidine 463 in the S5-P linker (turret) increases the rate of depolarization-induced inactivation and decreases the peak current amplitude. In this study, we examined how amino acid substitutions that altered the physico-chemical properties of the side chain at position 463 affected slow inactivation and then used the substituted cysteine accessibility method (SCAM) to probe the turret region (E456-P468) to determine whether residue 463 was unique in its ability to modulate the macroscopic current. Substitutions at position 463 of small, neutral (H463G and H463A) or large, charged (H463R, H463K, and H463E) side groups accelerated inactivation and induced a dependency of the current amplitude on the external potassium concentration. When cysteine substitutions were made in the distal turret (T462C-P468C), modification with either the positively charged [2-(trimethylammonium)ethyl] methanethiosulfonate bromide (MTSET) or negatively charged sodium (2-sulfonatoethyl) methanethiosulfonate reagent irreversibly inhibited current. This inhibition could be antagonized either by the R487V mutation (homologous to T449V in Shaker) or by raising the external potassium concentration, suggesting that current inhibition by MTS reagents resulted from an enhancement of inactivation. These results imply that protonation of residue 463 does not modulate inactivation solely by an electrostatic interaction with residues near the pore mouth, as proposed by others, and that residue 463 is part of a group of residues within the Kv1.5 turret that can modulate P/C-type inactivation.

  5. Mycobacteria inactivation using Engineered Water Nanostructures (EWNS)

    PubMed Central

    Pyrgiotakis, Georgios; McDevitt, James; Gao, Ya; Branco, Alan; Eleftheriadou, Mary; Lemos, Bernardo; Nardell, Edward; Demokritou, Philip

    2015-01-01

    Airborne transmitted pathogens such as Mycobacterium tuberculosis (Mtb) cause serious, often fatal infectious disease with enormous global health implications. Due to their unique cell wall and slow growth, mycobacteria are among the most resilient microbial forms. Herein we evaluate the ability of an emerging, chemical-free, nanotechnology-based method to inactivate M. parafortuitum (Mtb surrogate). This method is based on the transformation of atmospheric water vapor into engineered water nano-structures (EWNS) via electrospray. We demonstrate that the EWNS can interact with and inactivate airborne mycobacteria, reducing their concentration levels significantly. Additionally, EWNS can inactivate M. parafortuitum on surfaces eight times faster than the control. The mechanism of mycobacteria inactivation was also investigated in this study. It was demonstrated that the EWNS effectively deliver the reactive oxygen species, encapsulated during the electrospray process, to the bacteria oxidizing their cell membrane resulting into inactivation. Overall, this is a method with the potential to become an effective intervention technology in the battle against airborne infections. From the Clinical Editor This study demonstrates the feasibility of mycobacterium inactivation in airborne form or on contact surfaces using electrospray activated water nano-structures. Given that the method is free of toxic chemicals, this might become an important tool in the prevention of mycobacterial infections, which are notoriously hard to treat. PMID:24632246

  6. Photodynamic-induced inactivation of Propionibacterium acnes

    NASA Astrophysics Data System (ADS)

    Koenig, Karsten; Teschke, M.; Eick, Stephen G.; Pfister, W.; Meyer, Herbert; Halbhuber, Karl-Juergen

    1998-05-01

    We report on photodynamically induced inactivation of the skin bacterium Propionibacterium acnes (P. acnes) using endogenous as well as exogenous photosensitizers and red light sources. P. acnes is involved in the pathogenesis of the skin disease acne vulgaris. The skin bacterium is able to synthesize the metal-free fluorescent porphyrins protoporphyrin IX (PP) and coproporphyrin (CP) as shown by in situ spectrally-resolved detection of natural autofluorescence of human skin and bacteria colonies. These naturally occurring intracellular porphyrins act as efficient endogenous photosensitizers. Inactivation of P. acnes suspensions was achieved by irradiation with He-Ne laser light in the red spectral region (632.8 nm). We monitored the photodynamically-induced death of single bacteria using a fluorescent viability kit in combination with confocal laser scanning microscopy. In addition, the photo-induced inactivation was calculated by CFU (colony forming units) determination. We found 633 nm-induced inactivation (60 mW, 0.12 cm2 exposure area, 1 hour irradiation) of 72% in the case of non-incubated bacteria based on the destructive effect of singlet oxygen produced by red light excited endogenous porphyrins and subsequent energy transfer to molecular oxygen. In order to achieve a nearly complete inactivation within one exposure procedure, the exogenous photosensitizer Methylene Blue (Mb) was added. Far red exposure of Mb-labeled bacteria using a krypton ion laser at 647 nm and 676 nm resulted in 99% inactivation.

  7. Azide protection of bacteroides superoxide dismutases from inactivation by hydrogen peroxide

    SciTech Connect

    Barkley, K.B.; Gregory, E.M.

    1986-05-01

    The anaerobes Bacteroides fragilis, B. distasonis and B. thetaiotaomicron produce an iron-containing superoxide dismutase (FeSOD). These FeSODs are reversibly inhibited by 1 mM azide (NaN/sub 3/) and are irreversibly inactivated upon incubation with hydrogen peroxide (H/sub 2/O/sub 2/). H/sub 2/O/sub 2/ inactivation of the enzyme likely depends on a Fenton type reaction with the production of hydroxyl radical (OH). Addition of NaN/sub 3/ to the enzyme solution decreased the rate of inactivation by H/sub 2/O/sub 2/. After 20 minutes incubation of purified B. distasonis FeSOD with 2.5 mM H/sub 2/O/sub 2/, 61% of the initial enzymatic activity remained when 1 mM NaN/sub 3/ was also present compared with 29% activity without NaN/sub 3/. Similar results were seen with FeSOD from B. fragilis and B. thetaiotaomicron. Metal analyses of the native, peroxidized, and NaN/sub 3/ protected samples are consistent with loss of Fe from the enzyme upon peroxidation, but retention of Fe and enzymatic activity in the NaN/sub 3/ protected sample. Protection of FeSOD activity from H/sub 2/O/sub 2/ inactivation was dependent on NaN/sub 3/ concentration. Anionic hydroxyl radical scavengers, such as urate and xanthine did not significantly protect the enzyme. The results are consistent with binding of azide to the active site either preventing entry of H/sub 2/O/sub 2/ or altering Fe redox potential, preventing OH production.

  8. Evidence consistent with the requirement of cresolase activity for suicide inactivation of tyrosinase.

    PubMed

    Land, Edward J; Ramsden, Christopher A; Riley, Patrick A; Stratford, Michael R L

    2008-11-01

    Tyrosinase is a mono-oxygenase with a dinuclear copper catalytic center which is able to catalyze both the ortho-hydroxylation of monophenols (cresolase activity) and the oxidation of catechols (catecholase activity) yielding ortho-quinone products. Tyrosinases appear to have arisen early in evolution and are widespread in living organisms where they are involved in several processes, including antibiosis, adhesion of molluscs, the hardening of the exoskeleton of insects, and pigmentation. Tyrosinase is the principal enzyme of melanin formation in vertebrates and is of clinical interest because of the possible utilization of its activity for targeted treatment of malignant melanoma. Tyrosinase is characterised by an irreversible inactivation that occurs during the oxidation of catechols. In a recent publication we proposed a mechanism to account for this feature based on the ortho-hydroxylation of catecholic substrates, during which process Cu(II) is reduced to Cu(0) which no longer binds to the enzyme and is eliminated (reductive elimination). Since this process is dependent on cresolase activity of tyrosinase, a strong prediction of the proposed inactivation mechanism is that it will not be exhibited by enzymes lacking cresolase activity. We show that the catechol oxidase readily extracted from bananas (Musa cavendishii) is devoid of cresolase activity and that the kinetics of catechol oxidation do not exhibit inactivation. We also show that a species with the molecular mass of the putative cresolase oxidation product is formed during tyrosinase oxidation of 4-methylcatechol. The results presented are entirely consistent with our proposed mechanism to account for suicide-inactivation of tyrosinase.

  9. In Situ TEM Study of Reversible and Irreversible Electroforming in Pt/Ti:NiO/Pt Heterostructures

    SciTech Connect

    D'Aquila, Kenneth; Liu, Yuzi; Iddir, Hakim; Petford-Long, Amanda K.

    2015-05-01

    Experimental verification of the microscopic origin of resistance switching in metal/oxide/metal heterostructures is needed for applications in non-volatile memory and neuromorphic computing. Numerous reports suggest that resistance switching in NiO is caused by local reduction of the oxide layer into nanoscale conducting filaments, but few reports have shown experimental evidence correlating electroforming with site-specific changes in composition. We have investigated the mechanisms of reversible and irreversible electroforming in 250–500 nm wide pillars patterned from a single Ta/Ti/Pt/Ti-doped NiO/Pt/Ta heterostructure and have shown that these can coexist within a single sample. We performed in situ transmission electron microscopy (TEM) electroform- ing and switching on each pillar to correlate the local electron transport behavior with microstructure and composition in each pillar. DFT calculations fitted to electron energy loss spectroscopy data showed that the Ti-doped NiO layer is partially reduced after reversible electroforming, with the formation of oxygen vacancies ordered into lines in the <110> direction. However, under the same probing conditions, adjacent pillars show irreversible electroforming caused by electromigration of metallic Ta to form a single bridge across the oxide layer. We propose that the different electroforming behaviors are related to microstructural variations across the sample and may lead to switching variability.

  10. Inertial effects during irreversible meniscus reconfiguration in angular pores

    NASA Astrophysics Data System (ADS)

    Ferrari, Andrea; Lunati, Ivan

    2014-12-01

    In porous media, the dynamics of the invading front between two immiscible fluids is often characterized by abrupt reconfigurations caused by local instabilities of the interface. As a prototype of these phenomena we consider the dynamics of a meniscus in a corner as it can be encountered in angular pores. We investigate this process in detail by means of direct numerical simulations that solve the Navier-Stokes equations in the pore space and employ the Volume of Fluid method (VOF) to track the evolution of the interface. We show that for a quasi-static displacement, the numerically calculated surface energy agrees well with the analytical solutions that we have derived for pores with circular and square cross sections. However, the spontaneous reconfigurations are irreversible and cannot be controlled by the injection rate: they are characterized by the amount of surface energy that is spontaneously released and transformed into kinetic energy. The resulting local velocities can be orders of magnitude larger than the injection velocity and they induce damped oscillations of the interface that possess their own time scales and depend only on fluid properties and pore geometry. In complex media (we consider a network of cubic pores) reconfigurations are so frequent and oscillations last long enough that increasing inertial effects leads to a different fluid distribution by influencing the selection of the next pore to be invaded. This calls into question simple pore-filling rules based only on capillary forces. Also, we demonstrate that inertial effects during irreversible reconfigurations can influence the work done by the external forces that is related to the pressure drop in Darcy's law. This suggests that these phenomena have to be considered when upscaling multiphase flow because local oscillations of the menisci affect macroscopic quantities and modify the constitutive relationships to be used in macro-scale models. These results can be extrapolated to other

  11. Evaluation of irreversible JPEG compression for a clinical ultrasound practice.

    PubMed

    Persons, Kenneth R; Hangiandreou, Nicholas J; Charboneau, Nicholas T; Charboneau, J; James, E; Douglas, Bruce R; Salmon, Ann P; Knudsen, John M; Erickson, Bradley J

    2002-03-01

    A prior ultrasound study indicated that images with low to moderate levels of JPEG and wavelet compression were acceptable for diagnostic purposes. The purpose of this study is to validate this prior finding using the Joint Photographic Experts Group (JPEG) baseline compression algorithm, at a compression ratio of approximately 10:1, on a sufficiently large number of grayscale and color ultrasound images to attain a statistically significant result. The practical goal of this study is to determine if it is feasible for radiologists to use irreversibly compressed images as an integral part of the day to day ultrasound practice (ie, perform primary diagnosis with, and store irreversibly compressed images in the ultrasound PACS archive). In this study, 5 Radiologists were asked to review 300 grayscale and color static ultrasound images selected from 4 major anatomic groups. Each image was compressed and decompressed using the JPEG baseline compression algorithm at a fixed quality factor resulting in an average compression ratio of approximately 9:1. The images were presented in pairs (original and compressed) in a blinded fashion on a PACS workstation in the ultrasound reading areas, and radiologists were asked to pick which image they preferred in terms of diagnostic utility and their degree of certainty (on a scale from 1 to 4). Of the 1499 total readings, 50.17% (95% confidence intervals at 47.6%, and 52.7%) indicated a preference for the original image in the pair, and 49.83% (95% confidence intervals at 47.3%, and 52.0%) indicated a preference for the compressed image. These findings led the authors to conclude that static color and gray-scale ultrasound images compressed with JPEG at approximately 9:1 are statistically indistinguishable from the originals for primary diagnostic purposes. Based on the authors laboratory experience with compression and the results of this and other prior studies, JPEG compression is now being applied to all ultrasound images in

  12. Physicochemical stability and inactivation of human and simian rotaviruses

    SciTech Connect

    Meng, Z.D.; Birch, C.; Heath, R.; Gust, I.

    1987-04-01

    The effects of various physical and chemical treatments on the stability of a human serotype 1 rotavirus and simian agent 11 (SA11) were compared by using a fluorescence focus assay. The infectivity of both strains was retained after storage at room temperature for 14 days, 4 degree C for 22 days, and -20 degree C for 32 days; lyophilization; and treatment at pH 3 to 11. Both viruses were inactivated at pH 12, as was the human virus at pH 2, although this pH resulted in only partial inactivation of SA11. The human virus also appeared to be more sensitive than SA11 to the action of ether and chloroform. The infectivity of both viruses was lost after UV irradiation for 15 min and after treatment with 8% formaldehyde for 5 min, 70% (vol/vol) ethanol for 30 min, and 2% lysol, 2% phenol, and 1% H/sub 2/O/sub 2/ for 1 h each.

  13. Virus inactivation studies using ion beams, electron and gamma irradiation

    NASA Astrophysics Data System (ADS)

    Smolko, Eduardo E.; Lombardo, Jorge H.

    2005-07-01

    Known methods of virus inactivation are based on the chemical action of some substances such as acetylethylenimine, betapropiolactone, glycidalaldehyde, formaldehyde, etc. In such a process, the viral suspension should be kept at room or higher temperatures for 24-48 h. Under these conditions, physical and chemical agents act to degrade the virus antigenic proteins. On the contrary with ionizing radiations at low temperatures, the treatment does not cause such degradation allowing the study of different viral functions. In this work, particle (α, d and ß) and γ irradiations were used for partial and total inactivation of Foot and Mouth Disease Virus (FMDV), Rauscher Leukemia Virus (RLV) and Herpes Simplex Virus (HSV). Obtention of the D37 dose from survival curves and the application of the target theory, permitted the determination of molecular weight of the nucleic acid genomes, EBR values and useful information for vaccine preparation. For RLV virus, a two target model of the RNA genome was deduced in accordance with biological information while from data from the literature and our own work on the structure of the scrapie prion, considering the molecular weight obtained by application of the theory, a new model for prion replication is presented, based on a trimer molecule.

  14. Vaccine-associated enhanced respiratory disease is influenced by hemagglutinin and neuraminidase in whole inactivated influenza virus vaccines

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Multiple subtypes and many antigenic variants of influenza A virus (IAV) co-circulate in swine in the USA, complicating effective use of commercial vaccines to control disease and transmission. Whole inactivated virus (WIV) vaccines may provide partial protection against IAV with substantial antigen...

  15. Irreversible inhibitors of the 3C protease of Coxsackie virus through templated assembly of protein-binding fragments

    NASA Astrophysics Data System (ADS)

    Becker, Daniel; Kaczmarska, Zuzanna; Arkona, Christoph; Schulz, Robert; Tauber, Carolin; Wolber, Gerhard; Hilgenfeld, Rolf; Coll, Miquel; Rademann, Jörg

    2016-09-01

    Small-molecule fragments binding to biomacromolecules can be starting points for the development of drugs, but are often difficult to detect due to low affinities. Here we present a strategy that identifies protein-binding fragments through their potential to induce the target-guided formation of covalently bound, irreversible enzyme inhibitors. A protein-binding nucleophile reacts reversibly with a bis-electrophilic warhead, thereby positioning the second electrophile in close proximity of the active site of a viral protease, resulting in the covalent de-activation of the enzyme. The concept is implemented for Coxsackie virus B3 3C protease, a pharmacological target against enteroviral infections. Using an aldehyde-epoxide as bis-electrophile, active fragment combinations are validated through measuring the protein inactivation rate and by detecting covalent protein modification in mass spectrometry. The structure of one enzyme-inhibitor complex is determined by X-ray crystallography. The presented warhead activation assay provides potent non-peptidic, broad-spectrum inhibitors of enteroviral proteases.

  16. Irreversible inhibitors of the 3C protease of Coxsackie virus through templated assembly of protein-binding fragments

    PubMed Central

    Becker, Daniel; Kaczmarska, Zuzanna; Arkona, Christoph; Schulz, Robert; Tauber, Carolin; Wolber, Gerhard; Hilgenfeld, Rolf; Coll, Miquel; Rademann, Jörg

    2016-01-01

    Small-molecule fragments binding to biomacromolecules can be starting points for the development of drugs, but are often difficult to detect due to low affinities. Here we present a strategy that identifies protein-binding fragments through their potential to induce the target-guided formation of covalently bound, irreversible enzyme inhibitors. A protein-binding nucleophile reacts reversibly with a bis-electrophilic warhead, thereby positioning the second electrophile in close proximity of the active site of a viral protease, resulting in the covalent de-activation of the enzyme. The concept is implemented for Coxsackie virus B3 3C protease, a pharmacological target against enteroviral infections. Using an aldehyde-epoxide as bis-electrophile, active fragment combinations are validated through measuring the protein inactivation rate and by detecting covalent protein modification in mass spectrometry. The structure of one enzyme–inhibitor complex is determined by X-ray crystallography. The presented warhead activation assay provides potent non-peptidic, broad-spectrum inhibitors of enteroviral proteases. PMID:27677239

  17. Obatoclax interacts synergistically with the irreversible proteasome inhibitor carfilzomib in GC- and ABC- DLBCL cells in vitro and in vivo

    PubMed Central

    Dasmahapatra, Girija; Lembersky, Dmitry; Son, Minkyeong P.; Patel, Hiral; Peterson, Derick; Attkisson, Elisa; Fisher, Richard I.; Friedberg, Jonathan W.; Dent, Paul; Grant, Steven

    2013-01-01

    Interactions between the the irreversible proteasome inhibitor carfilzomib (CFZ) and the pan-BH3 mimetic obatoclax (Obato) were examined in GC- and ABC-DLBCL cells. Co-treatment with minimally toxic concentrations of CFZ (i.e., 2–6 nM) and sub-toxic concentrations of obato (0.05–2.0μM) synergistically increased apoptosis in multiple DLBCL cell lines and increased lethality toward primary human DLBCL but not normal CD34+ cells. Synergistic interactions were associated with sharp increases in caspase-3 activation, PARP cleavage, phospho-JNK induction, up-regulation of Noxa, and AKT dephosphorylation. Combined treatment also diminished CFZ-mediated Mcl-1 up-regulation while immunoprecipitation analysis revealed reduced associations between Bak and Mcl-1/Bcl-xL, and Bim and Mcl-1. The CFZ/Obato regimen triggered translocation, conformational change and dimerization of Bax and activation of Bak. Genetic interruption of JNK and Noxa by shRNA knockdown, ectopic Mcl-1 expression, or enforced activation of AKT significantly attenuated CFZ/Obato-mediated apoptosis. Notably, co-administration of CFZ/Obato sharply increased apoptosis in multiple bortezomib-resistant DLBCL models. Finally, in vivo administration of CFZ and Obato to mice inoculated with SUDHL4 cells substantially suppressed tumor growth, activated JNK, inactivated AKT, and increased survival compared to the effects of single agent treatment. Together, these findings argue that a strategy combining CFZ and Obato warrants attention in DLBCL. PMID:22411899

  18. The Impact of Uncertainty and Irreversibility on Investments in Online Learning

    ERIC Educational Resources Information Center

    Oslington, Paul

    2004-01-01

    Uncertainty and irreversibility are central to online learning projects, but have been neglected in the existing educational cost-benefit analysis literature. This paper builds some simple illustrative models of the impact of irreversibility and uncertainty, and shows how different types of cost and demand uncertainty can have substantial impacts…

  19. Flux creep and irreversibility line in high-temperature oxide superconductors

    SciTech Connect

    Matsushita, T.; Fujiyoshi, T.; Toko, K.; Yamafuji, K. )

    1990-05-14

    The irreversibility line in high-temperature oxide superconductors is theoretically investigated from a viewpoint of dependence on the flux-pinning strength and a general relation between the effective pinning potential and the critical current density is derived. It is shown that the irreversibility magnetic field at 77 K in strongly pinned oxide superconductors is sufficiently high for application.

  20. A minimal dissipation type-based classification in irreversible thermodynamics and microeconomics

    NASA Astrophysics Data System (ADS)

    Tsirlin, A. M.; Kazakov, V.; Kolinko, N. A.

    2003-10-01

    We formulate the problem of finding classes of kinetic dependencies in irreversible thermodynamic and microeconomic systems for which minimal dissipation processes belong to the same type. We show that this problem is an inverse optimal control problem and solve it. The commonality of this problem in irreversible thermodynamics and microeconomics is emphasized.

  1. State-dependent inactivation of the Kv3 potassium channel.

    PubMed Central

    Marom, S; Levitan, I B

    1994-01-01

    Inactivation of Kv3 (Kv1.3) delayed rectifier potassium channels was studied in the Xenopus oocyte expression system. These channels inactivate slowly during a long depolarizing pulse. In addition, inactivation accumulates in response to a series of short depolarizing pulses (cumulative inactivation), although no significant inactivation occurs within each short pulse. The extent of cumulative inactivation does not depend on the voltage during the depolarizing pulse, but it does vary in a biphasic manner as a function of the interpulse duration. Furthermore, the rate of cumulative inactivation is influenced by changing the rate of deactivation. These data are consistent with a model in which Kv3 channel inactivation is a state-dependent and voltage-independent process. Macroscopic and single channel experiments indicate that inactivation can occur from a closed (silent) state before channel opening. That is, channels need not open to inactivate. The transition that leads to the inactivated state from the silent state is, in fact, severalfold faster then the observed inactivation of current during long depolarizing pulses. Long pulse-induced inactivation appears to be slow, because its rate is limited by the probability that channels are in the open state, rather than in the silent state from which they can inactivate. External potassium and external calcium ions alter the rates of cumulative and long pulse-induced inactivation, suggesting that antagonistic potassium and calcium binding steps are involved in the normal gating of the channel. PMID:7948675

  2. Irreversible Entropy Production in Two-Phase Mixing Layers

    NASA Technical Reports Server (NTRS)

    Okongo, Nora

    2003-01-01

    This report presents a study of dissipation (irreversible production of entropy) in three-dimensional, temporal mixing layers laden with evaporating liquid drops. The purpose of the study is to examine the effects of evaporating drops on the development of turbulent features in flows. Direct numerical simulations were performed to analyze transitional states of three mixing layers: one without drops, and two that included drops at different initial mass loadings. Without drops, the dissipation is essentially due to viscous effects. It was found that in the presence of drops, the largest contribution to dissipation was made by heating and evaporation of the drops, and that at large length scales, this contribution is positive (signifying that the drops reduce turbulence), while at small scales, this contribution is negative (the drops increase turbulence). The second largest contribution to dissipation was found to be associated with the chemical potential, which leads to an increase in turbulence at large scales and a decrease in turbulence at small scales. The next smaller contribution was found to be that of viscosity. The fact that viscosity effects are only third in order of magnitude in the dissipation is in sharp contrast to the situation for the mixing layer without the drops. The next smaller contribution - that of the drag and momentum of the vapor from the drops - was found to be negative at lower mass loading but to become positive at higher mass loading.

  3. Electrical conductivity changes during irreversible electroporation treatment of brain cancer.

    PubMed

    Garcia, Paulo A; Rossmeisl, John H; Davalos, Rafael V

    2011-01-01

    Irreversible electroporation (IRE) is a new minimally invasive technique to kill tumors and other undesirable tissue in a non-thermal manner. During an IRE treatment, a series of short and intense electric pulses are delivered to the region of interest to destabilize the cell membranes in the tissue and achieve spontaneous cell death. The alteration of the cellular membrane results in a dramatic increase in electrical conductivity during IRE as in other electroporation-based-therapies. In this study, we performed the planning and execution of an IRE brain cancer treatment using MRI reconstructions of the tumor and a multichannel array that served as a stereotactic fiducial and electrode guide. Using the tumor reconstructions within our numerical simulations, we developed equations relating the increase in tumor conductivity to calculated currents and volumes of tumor treated with IRE. We also correlated the experimental current measured during the procedure to an increase in tumor conductivity ranging between 3.42-3.67 times the baseline conductivity, confirming the physical phenomenon that has been detected in other tissues undergoing similar electroporation-based treatments. PMID:22254416

  4. Time-dependent, irreversible entropy production and geodynamics.

    PubMed

    Regenauer-Lieb, Klaus; Karrech, Ali; Chua, Hui Tong; Horowitz, Franklin G; Yuen, Dave

    2010-01-13

    We present an application of entropy production as an abstraction tool for complex processes in geodynamics. Geodynamic theories are generally based on the principle of maximum dissipation being equivalent to the maximum entropy production. This represents a restriction of the second law of thermodynamics to its upper bound. In this paper, starting from the equation of motion, the first law of thermodynamics and decomposition of the entropy into reversible and irreversible terms,(1) we come up with an entropy balance equation in an integral form. We propose that the extrema of this equation give upper and lower bounds that can be used to constrain geodynamics solutions. This procedure represents an extension of the classical limit analysis theory of continuum mechanics, which considers only stress and strain rates. The new approach, however, extends the analysis to temperature-dependent problems where thermal feedbacks can play a significant role. We apply the proposed procedure to a simple convective/conductive heat transfer problem such as in a planetary system. The results show that it is not necessary to have a detailed knowledge of the material parameters inside the planet to derive upper and lower bounds for self-driven heat transfer processes. The analysis can be refined by considering precise dissipation processes such as plasticity and viscous creep. PMID:19948557

  5. Percolation of heteronuclear dimers irreversibly deposited on square lattices

    NASA Astrophysics Data System (ADS)

    Gimenez, M. C.; Ramirez-Pastor, A. J.

    2016-09-01

    The percolation problem of irreversibly deposited heteronuclear dimers on square lattices is studied. A dimer is composed of two segments, and it occupies two adjacent adsorption sites. Each segment can be either a conductive segment (segment type A ) or a nonconductive segment (segment type B ). Three types of dimers are considered: A A , B B , and A B . The connectivity analysis is carried out by accounting only for the conductive segments (segments type A ). The model offers a simplified representation of the problem of percolation of defective (nonideal) particles, where the presence of defects in the system is simulated by introducing a mixture of conductive and nonconductive segments. Different cases were investigated, according to the sequence of deposition of the particles, the types of dimers involved in the process, and the degree of alignment of the deposited objects. By means of numerical simulations and finite-size scaling analysis, the complete phase diagram separating a percolating from a nonpercolating region was determined for each case. Finally, the consistency of our results was examined by comparing with previous data in the literature for linear k -mers (particles occupying k adjacent sites) with defects.

  6. Non-Equilibrium Critical Behavior: An Extended Irreversible Thermodynamics Approach

    NASA Astrophysics Data System (ADS)

    Hernández-Lemus, Enrique; García-Colín, Leopoldo S.

    2006-11-01

    Critical phenomena in non-equilibrium systems have been studied by means of a wide variety of theoretical and experimental approaches. Mode-coupling, renormalization group, complex Lie algebras and diagrammatic techniques are some of the usual theoretical tools. Experimental studies include light and inelastic neutron scattering, X-ray photon correlation spectroscopy, microwave interferometry and several other techniques. Nevertheless, no conclusive treatment has been developed from the basic principles of a thermodynamic theory of irreversible processes. We have developed a formalism in which we obtain correlation functions as field averages of the associated functions. By applying such formalism, we attempt to find out whether the resulting correlation functions will inherit the mathematical properties (integrability, generalized homogeneity, scaling laws) of its parent potentials, and we also use these correlation functions to study the behavior of macroscopic systems far from equilibrium, especially in the neighborhood of critical points or dynamic phase transitions. As a working example, we will consider the mono-critical behavior of a non-equilibrium binary fluid mixture close to its consolute point.

  7. Scaling Law for Irreversible Entropy Production in Critical Systems

    NASA Astrophysics Data System (ADS)

    Hoang, Danh-Tai; Prasanna Venkatesh, B.; Han, Seungju; Jo, Junghyo; Watanabe, Gentaro; Choi, Mahn-Soo

    2016-06-01

    We examine the Jarzynski equality for a quenching process across the critical point of second-order phase transitions, where absolute irreversibility and the effect of finite-sampling of the initial equilibrium distribution arise in a single setup with equal significance. We consider the Ising model as a prototypical example for spontaneous symmetry breaking and take into account the finite sampling issue by introducing a tolerance parameter. The initially ordered spins become disordered by quenching the ferromagnetic coupling constant. For a sudden quench, the deviation from the Jarzynski equality evaluated from the ideal ensemble average could, in principle, depend on the reduced coupling constant ε0 of the initial state and the system size L. We find that, instead of depending on ε0 and L separately, this deviation exhibits a scaling behavior through a universal combination of ε0 and L for a given tolerance parameter, inherited from the critical scaling laws of second-order phase transitions. A similar scaling law can be obtained for the finite-speed quench as well within the Kibble-Zurek mechanism.

  8. Scaling Law for Irreversible Entropy Production in Critical Systems

    PubMed Central

    Hoang, Danh-Tai; Prasanna Venkatesh, B.; Han, Seungju; Jo, Junghyo; Watanabe, Gentaro; Choi, Mahn-Soo

    2016-01-01

    We examine the Jarzynski equality for a quenching process across the critical point of second-order phase transitions, where absolute irreversibility and the effect of finite-sampling of the initial equilibrium distribution arise in a single setup with equal significance. We consider the Ising model as a prototypical example for spontaneous symmetry breaking and take into account the finite sampling issue by introducing a tolerance parameter. The initially ordered spins become disordered by quenching the ferromagnetic coupling constant. For a sudden quench, the deviation from the Jarzynski equality evaluated from the ideal ensemble average could, in principle, depend on the reduced coupling constant ε0 of the initial state and the system size L. We find that, instead of depending on ε0 and L separately, this deviation exhibits a scaling behavior through a universal combination of ε0 and L for a given tolerance parameter, inherited from the critical scaling laws of second-order phase transitions. A similar scaling law can be obtained for the finite-speed quench as well within the Kibble-Zurek mechanism. PMID:27277558

  9. Prochloraz causes irreversible masculinization of zebrafish (Danio rerio).

    PubMed

    Baumann, Lisa; Knörr, Susanne; Keiter, Susanne; Nagel, Tina; Segner, Helmut; Braunbeck, Thomas

    2015-11-01

    The aim of the present study was to investigate the persistence of endocrine effects by prochloraz, a fungicide known to have multiple effects on the endocrine system of vertebrates. Since discontinuous exposure is particularly relevant in aquatic ecosystems, an exposure scenario with an exposure phase and a subsequent recovery period was chosen to assess the potential for reversibility of effects by prochloraz on the sexual development of zebrafish (Danio rerio). Zebrafish were exposed to different concentrations of prochloraz (10-300 μg/L) until 60 days post hatch (dph), which includes the period of sexual differentiation. For the subsequent 40 days, fish were either held in clean water for depuration or under further continuous exposure. Histological investigations of the gonads revealed persistent effects on sexual differentiation. The sex ratio was skewed towards males and significantly more intersex individuals were found after exposure to prochloraz at 60 dph. No intersex fish, but masculinized sex ratios were still present after the depuration period, documenting that prochloraz irreversibly affects the sexual development of zebrafish. PMID:25163568

  10. Use of irreversible electroporation in unresectable pancreatic cancer

    PubMed Central

    2015-01-01

    Irreversible electroporation is a non-thermal injury ablative modality that has been in clinical use since 2008 in the treatment of locally advanced soft tissue tumors. It has been reported to be utilized intraoperatively, laparoscopically or percutaneously. The method of action of IRE relies on a high voltage (maximum 3,000 volts) small microsecond pulse lengths (70 to 90 microseconds) to induce cell membrane porosity which leads to slow/protracted cell death over time. One of the largest unmet needs in oncology that IRE has been utilized is in locally advanced (stage III) pancreatic cancer. Recent studies have demonstrated the safety and palliation with encouraging improvement in overall survival. Its inherent limitation still remains tissue heterogeneity and the unique settings based on tumor histology and prior induction therapy. There remains a high technical demand of the end-user and the more extensive knowledge transfer which makes the learning curve longer in order to achieve appropriate and safe utilization. PMID:26151062

  11. Irreversible electroporation of locally advanced pancreatic neck/body adenocarcinoma

    PubMed Central

    2015-01-01

    Objective Irreversible electroporation (IRE) of locally advanced pancreatic adenocarcinoma of the neck has been used to palliate appropriate stage 3 pancreatic cancers without evidence of metastasis and who have undergone appropriate induction therapy. Currently there has not been a standardized reported technique for pancreatic mid-body tumors for patient selection and intra-operative technique. Patients Subjects are patients with locally advanced pancreatic adenocarcinoma of the body/neck who have undergone appropriate induction chemotherapy for a reasonable duration. Main outcome measures Technique of open IRE of locally advanced pancreatic adenocarcinoma of the neck/body is described, with the emphasis on intra-operative ultrasound and intra-operative electroporation management. Results The technique of open IRE of the pancreatic neck/body with bracketing of the celiac axis and superior mesenteric artery with continuous intraoperative ultrasound imaging and consideration of intraoperative navigational system is described. Conclusions IRE of locally advanced pancreatic adenocarcinoma of the body/neck is feasible for appropriate patients with locally advanced unresectable pancreatic cancer. PMID:26029461

  12. Scaling Law for Irreversible Entropy Production in Critical Systems.

    PubMed

    Hoang, Danh-Tai; Prasanna Venkatesh, B; Han, Seungju; Jo, Junghyo; Watanabe, Gentaro; Choi, Mahn-Soo

    2016-01-01

    We examine the Jarzynski equality for a quenching process across the critical point of second-order phase transitions, where absolute irreversibility and the effect of finite-sampling of the initial equilibrium distribution arise in a single setup with equal significance. We consider the Ising model as a prototypical example for spontaneous symmetry breaking and take into account the finite sampling issue by introducing a tolerance parameter. The initially ordered spins become disordered by quenching the ferromagnetic coupling constant. For a sudden quench, the deviation from the Jarzynski equality evaluated from the ideal ensemble average could, in principle, depend on the reduced coupling constant ε0 of the initial state and the system size L. We find that, instead of depending on ε0 and L separately, this deviation exhibits a scaling behavior through a universal combination of ε0 and L for a given tolerance parameter, inherited from the critical scaling laws of second-order phase transitions. A similar scaling law can be obtained for the finite-speed quench as well within the Kibble-Zurek mechanism. PMID:27277558

  13. Distribution function approach to irreversible adsorption of interacting colloidal particles

    NASA Astrophysics Data System (ADS)

    Faraudo, Jordi; Bafaluy, Javier

    2000-01-01

    A statistical-mechanical description of the irreversible adsorption of interacting colloidal particles is developed. Our approach describes in a consistent way the interaction of particles from the bulk with adsorbed particles during the transport process towards the adsorbing surface. The macroscopic physical quantities corresponding to the actual process are expressed as averages over simpler auxiliary processes which proceed in the presence of a fixed number n of adsorbed particles. The adsorption rate verifies a generalized Langmuir equation, in which the kinetic resistance (the inverse of the kinetic coefficient) is expressed as the sum of a diffusional resistance and a resistance due to interaction with adsorbed particles during the transport process (blocking effect). Contrary to previous approaches, the blocking effect is not due to geometrical exclusion, instead it measures how the transport from the bulk is affected by the adsorbed particles. From the general expressions obtained, we have derived coverage expansions for the adsorption rate and the surface correlation function. The theory is applied to the case of colloidal particles interacting through DLVO potentials. This form of the kinetic coefficient is shown to be in agreement with recent experimental results, in which RSA fails.

  14. The nineteenth century conflict between mechanism and irreversibility

    NASA Astrophysics Data System (ADS)

    van Strien, Marij

    2013-08-01

    The reversibility problem (better known as the reversibility objection) is usually taken to be an internal problem in the kinetic theory of gases, namely the problem of how to account for the second law of thermodynamics within this theory. Historically, it is seen as an objection that was raised against Boltzmann's kinetic theory of gases, which led Boltzmann to a statistical approach to the kinetic theory, culminating in the development of statistical mechanics. In this paper, I show that in the late nineteenth century, the reversibility problem had a much broader significance-it was widely discussed and certainly not only as an objection to Boltzmann's kinetic theory of gases. In this period, there was a conflict between mechanism and irreversibility in physics which was tied up with central issues in philosophy of science such as materialism, empiricism and the need for mechanistic foundations of physical theories, as well as with concerns about the heat death of the universe. I discuss how this conflict was handled by the major physicists of the period, such as Maxwell, Kelvin, Duhem, Poincaré, Mach and Planck, as well as by a number of lesser-known authors.

  15. Irreversible opinion spreading on scale-free networks

    NASA Astrophysics Data System (ADS)

    Candia, Julián

    2007-02-01

    We study the dynamical and critical behavior of a model for irreversible opinion spreading on Barabási-Albert (BA) scale-free networks by performing extensive Monte Carlo simulations. The opinion spreading within an inhomogeneous society is investigated by means of the magnetic Eden model, a nonequilibrium kinetic model for the growth of binary mixtures in contact with a thermal bath. The deposition dynamics, which is studied as a function of the degree of the occupied sites, shows evidence for the leading role played by hubs in the growth process. Systems of finite size grow either ordered or disordered, depending on the temperature. By means of standard finite-size scaling procedures, the effective order-disorder phase transitions are found to persist in the thermodynamic limit. This critical behavior, however, is absent in related equilibrium spin systems such as the Ising model on BA scale-free networks, which in the thermodynamic limit only displays a ferromagnetic phase. The dependence of these results on the degree exponent is also discussed for the case of uncorrelated scale-free networks.

  16. Generalized model and optimum performance of an irreversible quantum Brayton engine with spin systems.

    PubMed

    Wu, Feng; Chen, Lingen; Sun, Fengrui; Wu, Chih; Li, Qing

    2006-01-01

    The purpose of this paper is to establish a model of an irreversible quantum Brayton engine using many noninteracting spin systems as the working substance and consisting of two irreversible adiabatic and two isomagnetic field processes. The time evolution of the total magnetic moment M is determined by solving the generalized quantum master equation of an open system in the Heisenberg picture. The time of two irreversible adiabatic processes is considered based on finite-rate evolution. The relationship between the power output P and the efficiency eta for the irreversible quantum Brayton engine with spin systems is derived. The optimally operating region (or criteria) for the engine is determined. The influences of these important parameters on the performances (P and eta) of the engine are discussed. The results obtained herein will be useful for the further understanding and the selection of the optimal operating conditions for an irreversible quantum Brayton engine with spin systems.

  17. Macroscopic irreversibility and microscopic paradox: A Constructal law analysis of atoms as open systems

    PubMed Central

    Lucia, Umberto

    2016-01-01

    The relation between macroscopic irreversibility and microscopic reversibility is a present unsolved problem. Constructal law is introduced to develop analytically the Einstein’s, Schrödinger’s, and Gibbs’ considerations on the interaction between particles and thermal radiation (photons). The result leads to consider the atoms and molecules as open systems in continuous interaction with flows of photons from their surroundings. The consequent result is that, in any atomic transition, the energy related to the microscopic irreversibility is negligible, while when a great number of atoms (of the order of Avogadro’s number) is considered, this energy related to irreversibility becomes so large that its order of magnitude must be taken into account. Consequently, macroscopic irreversibility results related to microscopic irreversibility by flows of photons and amount of atoms involved in the processes. PMID:27762333

  18. Reanalysis of experiments to quantify irreversibility of pesticide sorption-desorption in soil.

    PubMed

    Suddaby, Laura A; Beulke, Sabine; van Beinum, Wendy; Celis, Rafael; Koskinen, William C; Brown, Colin D

    2013-03-01

    Previously published research used an isotope-exchange technique to measure irreversibility of pesticide sorption-desorption in soil. Results indicated significant irreversibility (6-51%) in sorption in five pesticide-soil systems measured over 72 h. Here, we propose a three-site model to reanalyze the experimental data. The model adds a slow but reversible binding on nonequilibrium sorption sites in addition to instantaneously reversible sites and irreversible sites. The model was able to match experimental data very closely, but only if irreversible sorption was assumed to be absent. Observed asymmetry in the binding of (12)C- and (14)C-pesticide was explained on the basis of nonattainment of sorption equilibrium over the study period. Results suggest that irreversible sorption may be less significant than previously considered with important implications for understanding the fate of pesticides applied to soil.

  19. Succination of proteins by fumarate: mechanism of inactivation of glyceraldehyde-3-phosphate dehydrogenase in diabetes.

    PubMed

    Blatnik, Matthew; Thorpe, Suzanne R; Baynes, John W

    2008-04-01

    S-(2-succinyl)cysteine (2SC) is a chemical modification of proteins formed by a Michael addition reaction between the Krebs cycle intermediate, fumarate, and thiol groups in protein--a process known as succination of protein. Succination causes irreversible inactivation of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in vitro. GAPDH was immunoprecipitated from muscle of diabetic rats, then analyzed by ultra-performance liquid chromatography-electrospray ionization-mass spectroscopy. Succination of GAPDH was increased in muscle of diabetic rats, and the extent of succination correlated strongly with the decrease in specific activity of the enzyme. We propose that 2SC is a biomarker of mitochondrial and oxidative stress in diabetes and that succination of GAPDH and other thiol proteins may provide the chemical link between glucotoxicity and the pathogenesis of diabetic complications.

  20. Partial (focal) seizure

    MedlinePlus

    ... Jacksonian seizure; Seizure - partial (focal); Temporal lobe seizure; Epilepsy - partial seizures ... Abou-Khalil BW, Gallagher MJ, Macdonald RL. Epilepsies. In: Daroff ... Practice . 7th ed. Philadelphia, PA: Elsevier; 2016:chap 101. ...

  1. Partial tooth gear bearings

    NASA Technical Reports Server (NTRS)

    Vranish, John M. (Inventor)

    2010-01-01

    A partial gear bearing including an upper half, comprising peak partial teeth, and a lower, or bottom, half, comprising valley partial teeth. The upper half also has an integrated roller section between each of the peak partial teeth with a radius equal to the gear pitch radius of the radially outwardly extending peak partial teeth. Conversely, the lower half has an integrated roller section between each of the valley half teeth with a radius also equal to the gear pitch radius of the peak partial teeth. The valley partial teeth extend radially inwardly from its roller section. The peak and valley partial teeth are exactly out of phase with each other, as are the roller sections of the upper and lower halves. Essentially, the end roller bearing of the typical gear bearing has been integrated into the normal gear tooth pattern.

  2. Mechanism of Multivalent Nanoparticle Encounter with HIV-1 for Potency Enhancement of Peptide Triazole Virus Inactivation*

    PubMed Central

    Rosemary Bastian, Arangassery; Nangarlia, Aakansha; Bailey, Lauren D.; Holmes, Andrew; Kalyana Sundaram, R. Venkat; Ang, Charles; Moreira, Diogo R. M.; Freedman, Kevin; Duffy, Caitlin; Contarino, Mark; Abrams, Cameron; Root, Michael; Chaiken, Irwin

    2015-01-01

    Entry of HIV-1 into host cells remains a compelling yet elusive target for developing agents to prevent infection. A peptide triazole (PT) class of entry inhibitor has previously been shown to bind to HIV-1 gp120, suppress interactions of the Env protein at host cell receptor binding sites, inhibit cell infection, and cause envelope spike protein breakdown, including gp120 shedding and, for some variants, virus membrane lysis. We found that gold nanoparticle-conjugated forms of peptide triazoles (AuNP-PT) exhibit substantially more potent antiviral effects against HIV-1 than corresponding peptide triazoles alone. Here, we sought to reveal the mechanism of potency enhancement underlying nanoparticle conjugate function. We found that altering the physical properties of the nanoparticle conjugate, by increasing the AuNP diameter and/or the density of PT conjugated on the AuNP surface, enhanced potency of infection inhibition to impressive picomolar levels. Further, compared with unconjugated PT, AuNP-PT was less susceptible to reduction of antiviral potency when the density of PT-competent Env spikes on the virus was reduced by incorporating a peptide-resistant mutant gp120. We conclude that potency enhancement of virolytic activity and corresponding irreversible HIV-1 inactivation of PTs upon AuNP conjugation derives from multivalent contact between the nanoconjugates and metastable Env spikes on the HIV-1 virus. The findings reveal that multispike engagement can exploit the metastability built into virus the envelope to irreversibly inactivate HIV-1 and provide a conceptual platform to design nanoparticle-based antiviral agents for HIV-1 specifically and putatively for metastable enveloped viruses generally. PMID:25371202

  3. Kinetics of Hydrothermal Inactivation of Endotoxins ▿

    PubMed Central

    Li, Lixiong; Wilbur, Chris L.; Mintz, Kathryn L.

    2011-01-01

    A kinetic model was established for the inactivation of endotoxins in water at temperatures ranging from 210°C to 270°C and a pressure of 6.2 × 106 Pa. Data were generated using a bench scale continuous-flow reactor system to process feed water spiked with endotoxin standard (Escherichia coli O113:H10). Product water samples were collected and quantified by the Limulus amebocyte lysate assay. At 250°C, 5-log endotoxin inactivation was achieved in about 1 s of exposure, followed by a lower inactivation rate. This non-log-linear pattern is similar to reported trends in microbial survival curves. Predictions and parameters of several non-log-linear models are presented. In the fast-reaction zone (3- to 5-log reduction), the Arrhenius rate constant fits well at temperatures ranging from 120°C to 250°C on the basis of data from this work and the literature. Both biphasic and modified Weibull models are comparable to account for both the high and low rates of inactivation in terms of prediction accuracy and the number of parameters used. A unified representation of thermal resistance curves for a 3-log reduction and a 3 D value associated with endotoxin inactivation and microbial survival, respectively, is presented. PMID:21193667

  4. Physiology and Pathophysiology of Sodium Channel Inactivation.

    PubMed

    Ghovanloo, M-R; Aimar, K; Ghadiry-Tavi, R; Yu, A; Ruben, P C

    2016-01-01

    Voltage-gated sodium channels are present in different tissues within the human body, predominantly nerve, muscle, and heart. The sodium channel is composed of four similar domains, each containing six transmembrane segments. Each domain can be functionally organized into a voltage-sensing region and a pore region. The sodium channel may exist in resting, activated, fast inactivated, or slow inactivated states. Upon depolarization, when the channel opens, the fast inactivation gate is in its open state. Within the time frame of milliseconds, this gate closes and blocks the channel pore from conducting any more sodium ions. Repetitive or continuous stimulations of sodium channels result in a rate-dependent decrease of sodium current. This process may continue until the channel fully shuts down. This collapse is known as slow inactivation. This chapter reviews what is known to date regarding, sodium channel inactivation with a focus on various mutations within each NaV subtype and with clinical implications. PMID:27586293

  5. Chromophore assisted laser inactivation of cellular proteins

    NASA Astrophysics Data System (ADS)

    Jay, Daniel G.; Wang, F. S.; Chang, H. Y.; Sydor, A. M.; Liao, J. C.

    1997-05-01

    A molecular understanding of biology requires that we establish the in situ functions of the proteins in cellular processes. To address this, we developed chromophore- assisted laser inactivation (CALI) for probing the in vivo function of proteins. CALI inactivates specific proteins in living cells by using non-blocking antibodies conjugated with malachite green (MG) dye. MG absorbs 620 nm laser light (which is not absorbed by cells) to generate short lived free radicals with limited range of oxidative damage (15 angstroms) around the dye. This inactivates the bound protein without significantly affecting its neighbors. CALI has been applied to 40 proteins and achieved specific inactivation in almost all those tested. We have developed micro-CALI which uses a focused laser beam (10 micrometers ) to acutely inactivate specific proteins within cells. We have used this to address the molecular mechanisms of neuronal growth cone motility and has implicated a diversity of proteins (e.g. molecular motors, cytoskeletal, and signaling molecules) in discrete steps of growth cone motility. We hope that micro-CALI will be a useful research tool for addressing dynamic processes in biology and medicine.

  6. Functional inactivation of lymphocytes by methylene blue with visible light.

    PubMed

    Zhang, Bo; Cheng, Zhenzhen; Mo, Qin; Wang, Li; Wang, Xun; Wu, Xiaofei; Jia, Yao; Huang, Yuwen

    2015-10-01

    Transfusion of allogeneic white blood cells (WBCs) may cause adverse reactions in immunocompromised recipients, including transfusion-associated graft-versus-host disease (TA-GVHD), which is often fatal and incurable. In this study, the in vitro effect of methylene blue with visible light (MB + L) treatment on lymphocyte proliferation and cytokine production was measured to investigate whether MB + L can be used to prevent immune reactions that result from transfused lymphocytes. WBCs and 3 μM of MB were mixed and transferred into medical PVC bags, which were then exposed to visible light. Gamma irradiation was conducted as a parallel positive control. The cells without treatment were used as untreated group. All the groups were tested for the ability of cell proliferation and cytokine production upon stimulation. After incubation with mitogen phytohemagglutinin (PHA) or plate-bound anti-CD3 plus anti-CD28, the proliferation of MB + L/gamma-irradiation treated lymphocytes was significantly inhibited (P < 0.01) as compared to the untreated ones; the proliferation inhibitive rate of the MB + L group was even higher than that of gamma-irradiated cells (73.77% ± 28.75% vs. 44.72% ± 38.20%). MB + L treated cells incubated up to 7 days with PHA also showed no significant proliferation. The levels of TNF-α, IFN-γ, IL-6, IL-8, IL-10 and IL-1β present in the supernatant of MB + L treated lymphocytes upon stimulation were significantly lower than those of untreated lymphocytes. These results demonstrated that MB + L treatment functionally and irreversibly inactivated lymphocytes by inhibiting lymphocyte proliferation and the production of cytokines. MB + L treatment might be a promising method for the prevention of adverse immune responses caused by WBCs. PMID:26295729

  7. Structural basis for the coupling between activation and inactivation gates in K+ channels

    PubMed Central

    Cuello, Luis G.; Jogini, Vishwanath; Cortes, D. Marien.; Pan, Albert C; Gagnon, Dominique G.; Dalmas, Olivier; Cordero-Morales, Julio F.; Chakrapani, Sudha; Roux, Benoit; Perozo, Eduardo

    2011-01-01

    The coupled interplay between activation and inactivation gating is a functional hallmark of K+ channels1,2. This coupling has been experimentally demonstrated from ion interaction effects3,4, cysteine accessibility1 and is associated with a well-defined boundary of energetically coupled residues2. The structure of KcsA in its fully open conformation, as well as four other partial openings, richly illustrates the structural basis of activation-inactivation gating5. Here, we have identified the mechanistic principles by which movements on the inner bundle gate trigger conformational changes at the selectivity filter, leading to the non-conductive C-type inactivated state. Analysis of a series of KcsA open structures suggests that as a consequence of the hinge bending and rotation of TM2, the aromatic ring of Phe103 tilts towards residues Thr74 and Thr75 in the pore helix as well as Ile100 in the neighboring subunit. This allows the network of hydrogen bonds among residues W67, E71, and D80 to destabilize the selectivity filter6,7, facilitating entry to its non-conductive conformation. Mutations at position 103, affect gating kinetics in a size-dependent way: small side chain substitutions F103A and F103C severely impair inactivation kinetics, while larger side chains (F103W) have more subtle effects. This suggests that the allosteric coupling between the inner helical bundle and the selectivity filter might rely on straightforward mechanical deformation propagated through a network of steric contacts. Average interactions calculated from molecular dynamics simulations show favourable open state interaction-energies between Phe103 and surrounding residues. Similar interactions were probed in the Shaker K-channel where inactivation was impaired in the mutant I470A. We propose that side chain rearrangements at position 103 mechanically couple activation and inactivation in KcsA and a variety of other K channels. PMID:20613845

  8. Carvacrol suppresses high pressure high temperature inactivation of Bacillus cereus spores.

    PubMed

    Luu-Thi, Hue; Corthouts, Jorinde; Passaris, Ioannis; Grauwet, Tara; Aertsen, Abram; Hendrickx, Marc; Michiels, Chris W

    2015-03-16

    The inactivation of bacterial spores generally proceeds faster and at lower temperatures when heat treatments are conducted under high pressure, and high pressure high temperature (HPHT) processing is, therefore, receiving an increased interest from food processors. However, the mechanisms of spore inactivation by HPHT treatment are poorly understood, particularly at moderately elevated temperature. In the current work, we studied inactivation of the spores of Bacillus cereus F4430/73 by HPHT treatment for 5 min at 600MPa in the temperature range of 50-100°C, using temperature increments of 5°C. Additionally, we investigated the effect of the natural antimicrobial carvacrol on spore germination and inactivation under these conditions. Spore inactivation by HPHT was less than about 1 log unit at 50 to 70°C, but gradually increased at higher temperatures up to about 5 log units at 100°C. DPA release and loss of spore refractility in the spore population were higher at moderate (≤65°C) than at high (≥70°C) treatment temperatures, and we propose that moderate conditions induced the normal physiological pathway of spore germination resulting in fully hydrated spores, while at higher temperatures this pathway was suppressed and replaced by another mechanism of pressure-induced dipicolinic acid (DPA) release that results only in partial spore rehydration, probably because spore cortex hydrolysis is inhibited. Carvacrol strongly suppressed DPA release and spore rehydration during HPHT treatment at ≤65°C and also partly inhibited DPA release at ≥65°C. Concomitantly, HPHT spore inactivation was reduced by carvacrol at 65-90°C but unaffected at 95-100°C.

  9. Irreversible Collective Migration of Cyanobacteria in Eutrophic Conditions

    PubMed Central

    Dervaux, Julien; Mejean, Annick; Brunet, Philippe

    2015-01-01

    In response to natural or anthropocentric pollutions coupled to global climate changes, microorganisms from aquatic environments can suddenly accumulate on water surface. These dense suspensions, known as blooms, are harmful to ecosystems and signicantly degrade the quality of water resources. In order to determine the physico-chemical parameters involved in their formation and quantitatively predict their appearance, we successfully reproduced irreversible cyanobacterial blooms in vitro. By combining chemical, biochemical and hydrodynamic evidences, we identify a mechanism, unrelated to the presence of internal gas vesicles, allowing the sudden collective upward migration in test tubes of several cyanobacterial strains (Microcystis aeruginosa PCC 7005, Microcystis aeruginosa PCC 7806 and Synechocystis sp. PCC 6803). The final state consists in a foamy layer of biomass at the air-liquid interface, in which micro-organisms remain alive for weeks, the medium lying below being almost completely depleted of cyanobacteria. These "laboratory blooms" start with the aggregation of cells at high ionic force in cyanobacterial strains that produce anionic extracellular polymeric substances (EPS). Under appropriate conditions of nutrients and light intensity, the high photosynthetic activity within cell clusters leads the dissolved oxygen (DO) to supersaturate and to nucleate into bubbles. Trapped within the EPS, these bubbles grow until their buoyancy pulls the biomass towards the free surface. By investigating a wide range of spatially homogeneous environmental conditions (illumination, salinity, cell and nutrient concentration) we identify species-dependent thresholds and timescales for bloom formation. We conclude on the relevance of such results for cyanobacterial bloom formation in the environment and we propose an ecient method for biomass harvesting in bioreactors. PMID:25799424

  10. Cold inactivation and dissociation into dimers of Escherichia coli tryptophanase and its W330F mutant form.

    PubMed

    Erez, T; Gdalevsky GYa; Torchinsky, Y M; Phillips, R S; Parola, A H

    1998-05-19

    The kinetics and mechanism of reversible cold inactivation of the tetrameric enzyme tryptophanase have been studied. Cold inactivation is shown to occur slowly in the presence of K+ ions and much faster in their absence. The W330F mutant tryptophanase undergoes rapid cold inactivation even in the presence of K+ ions. In all cases the inactivation is accompanied by a decrease of the coenzyme 420-nm CD and absorption peaks and a shift of the latter peak to shorter wavelengths. The spectral changes and the NaBH4 test indicate that cooling of tryptophanase leads to breaking of the internal aldimine bond and release of the coenzyme. HPLC analysis showed that the ensuing apoenzyme dissociates into dimers. The dissociation depends on the nature and concentration of anions in the buffer solution. It readily occurs at low protein concentrations in the presence of salting-in anions Cl-, NO3- and I-, whereas salting-out anions, especially HPO4(2-), hinder the dissociation. K+ ions do not influence the dissociation of the apoenzyme, but partially protect holotryptophanase from cold inactivation. Thus, the two processes, cold inactivation of tryptophanase and dissociation of its apoform into dimers exhibit different dependencies on K+ ions and anions.

  11. Role of oxyradicals in the inactivation of catalase by ozone

    SciTech Connect

    Whiteside, C.; Hassan, H.M. )

    1988-01-01

    The antioxidant enzymes, catalase and superoxide dismutase, are inactivated upon exposure to ozone. In this study, the mechanism of this inactivation was examined using catalase as a model system. The data show that the inactivation of catalase is dependent on ozone concentration, time of exposure, and pH. Loss of catalase activity is accompanied with loss of the heme spectra. Tiron, desferal-Mn, trolox-c, and pyruvate protect the enzyme against ozone inactivation. SOD is less effective due to its inactivation by ozone. On the other hand, alcohols do not provide significant protection. The data suggest the possible involvement of superoxide radicals in the inactivation of catalase by ozone.

  12. Irreversible Electroporation Near the Heart: Ventricular Arrhythmias Can Be Prevented With ECG Synchronization

    PubMed Central

    Deodhar, Ajita; Dickfeld, Timm; Single, Gordon W.; Hamilton, William C.; Thornton, Raymond H.; Sofocleous, Constantinos T.; Maybody, Majid; Gónen, Mithat; Rubinsky, Boris; Solomon, Stephen B.

    2013-01-01

    OBJECTIVE Irreversible electroporation is a nonthermal ablative tool that uses direct electrical pulses to create irreversible membrane pores and cell death. The ablation zone is surrounded by a zone of reversibly increased permeability; either zone can cause cardiac arrhythmias. Our purpose was to establish a safety profile for the use of irreversible electroporation close to the heart. MATERIALS and METHODS The effect of unsynchronized and synchronized (with the R wave on ECG) irreversible electroporation in swine lung and myocardium was studied in 11 pigs. Twelve lead ECG recordings were analyzed by an electrophysiologist for the presence of arrhythmia. Ventricular arrhythmias were categorized as major events. Minor events included all other dysrhythmias or ECG changes. Cardiac and lung tissue was submitted for histopathologic analysis. Electrical field modeling was performed to predict the distance from the applicators over which cells show electroporation-induced increased permeability. RESULTS At less than or equal to 1.7 cm from the heart, fatal (major) events occurred with all unsynchronized irreversible electroporation. No major and three minor events were seen with synchronized irreversible electroporation. At more than 1.7 cm from the heart, two minor events occurred with only unsynchronized irreversible electroporation. Electrical field modeling correlates well with the clinical results, revealing increased cell membrane permeability up to 1.7 cm away from the applicators. Complete lung ablation without intervening live cells was seen. No myocardial injury was seen. CONCLUSION Unsynchronized irreversible electroporation close to the heart can cause fatal ventricular arrhythmias. Synchronizing irreversible electroporation pulse delivery with absolute refractory period avoids significant cardiac arrhythmias. PMID:21343484

  13. Mesoscale modeling of irreversible volume growth in powders of anisotropic crystals

    SciTech Connect

    Gee, R; Maiti, A; Fried, L

    2006-05-05

    Careful thermometric analysis (TMA) on powders of micron-sized triamino-trinitrobenzene (TATB) crystallites are shown to display irreversible growth in volume when subjected to repeated cycles of heating and cooling. Such behavior is counter-intuitive to typical materials response to simulated annealing cycles in atomic-scale molecular dynamics. However, through coarse-grained simulations using a mesoscale Hamiltonian we quantitatively reproduce irreversible growth behavior in such powdered material. We demonstrate that irreversible growth happens only in the presence of intrinsic crystalline anisotropy, and is mediated by particles much smaller than the average crystallite size.

  14. Dermal and mucosal reactions to an antimicrobial irreversible hydrocolloid impression material: a clinical report.

    PubMed

    Baker, Philip S; Plummer, Kevin D; Parr, Gregory R; Parker, M Harry

    2006-03-01

    As an adjunct to infection control in dental impression procedures, several manufacturers have incorporated disinfectants into irreversible hydrocolloid impression materials. However, these compounds have been shown to be tissue irritants and capable of producing allergic reactions. An anonymous, self-administered questionnaire was distributed to 56 second-year dental students who had used an irreversible hydrocolloid containing a quaternary ammonium compound as an antimicrobial (Jeltrate Plus) to make impressions during a summer preclinical occlusion course. Within the limitations of this report, the incorporation of a quaternary ammonium compound into an irreversible hydrocolloid impression material resulted in a greater incidence of dermal and mucosal irritation.

  15. A Fingerprint Encryption Scheme Based on Irreversible Function and Secure Authentication

    PubMed Central

    Yu, Jianping; Zhang, Peng; Wang, Shulan

    2015-01-01

    A fingerprint encryption scheme based on irreversible function has been designed in this paper. Since the fingerprint template includes almost the entire information of users' fingerprints, the personal authentication can be determined only by the fingerprint features. This paper proposes an irreversible transforming function (using the improved SHA1 algorithm) to transform the original minutiae which are extracted from the thinned fingerprint image. Then, Chinese remainder theorem is used to obtain the biokey from the integration of the transformed minutiae and the private key. The result shows that the scheme has better performance on security and efficiency comparing with other irreversible function schemes. PMID:25873989

  16. A fingerprint encryption scheme based on irreversible function and secure authentication.

    PubMed

    Yang, Yijun; Yu, Jianping; Zhang, Peng; Wang, Shulan

    2015-01-01

    A fingerprint encryption scheme based on irreversible function has been designed in this paper. Since the fingerprint template includes almost the entire information of users' fingerprints, the personal authentication can be determined only by the fingerprint features. This paper proposes an irreversible transforming function (using the improved SHA1 algorithm) to transform the original minutiae which are extracted from the thinned fingerprint image. Then, Chinese remainder theorem is used to obtain the biokey from the integration of the transformed minutiae and the private key. The result shows that the scheme has better performance on security and efficiency comparing with other irreversible function schemes. PMID:25873989

  17. A fingerprint encryption scheme based on irreversible function and secure authentication.

    PubMed

    Yang, Yijun; Yu, Jianping; Zhang, Peng; Wang, Shulan

    2015-01-01

    A fingerprint encryption scheme based on irreversible function has been designed in this paper. Since the fingerprint template includes almost the entire information of users' fingerprints, the personal authentication can be determined only by the fingerprint features. This paper proposes an irreversible transforming function (using the improved SHA1 algorithm) to transform the original minutiae which are extracted from the thinned fingerprint image. Then, Chinese remainder theorem is used to obtain the biokey from the integration of the transformed minutiae and the private key. The result shows that the scheme has better performance on security and efficiency comparing with other irreversible function schemes.

  18. A Fragment-Based Method to Discover Irreversible Covalent Inhibitors of Cysteine Proteases

    PubMed Central

    2015-01-01

    A novel fragment-based drug discovery approach is reported which irreversibly tethers drug-like fragments to catalytic cysteines. We attached an electrophile to 100 fragments without significant alterations in the reactivity of the electrophile. A mass spectrometry assay discovered three nonpeptidic inhibitors of the cysteine protease papain. The identified compounds display the characteristics of irreversible inhibitors. The irreversible tethering system also displays specificity: the three identified papain inhibitors did not covalently react with UbcH7, USP08, or GST-tagged human rhinovirus 3C protease. PMID:24870364

  19. Inflammation Induces Irreversible Biophysical Changes in Isolated Nucleus Pulposus Cells

    PubMed Central

    Maidhof, Robert; Jacobsen, Timothy; Papatheodorou, Angelos; Chahine, Nadeen O.

    2014-01-01

    Intervertebral disc degeneration is accompanied by elevated levels of inflammatory cytokines that have been implicated in disease etiology and matrix degradation. While the effects of inflammatory stimulation on disc cell metabolism have been well-studied, their effects on cell biophysical properties have not been investigated. The hypothesis of this study is that inflammatory stimulation alters the biomechanical properties of isolated disc cells and volume responses to step osmotic loading. Cells from the nucleus pulposus (NP) of bovine discs were isolated and treated with either lipopolysaccharide (LPS), an inflammatory ligand, or with the recombinant cytokine TNF-α for 24 hours. We measured cellular volume regulation responses to osmotic loading either immediately after stimulation or after a 1 week recovery period from the inflammatory stimuli. Cells from each group were tested under step osmotic loading and the transient volume-response was captured via time-lapse microscopy. Volume-responses were analyzed using mixture theory framework to investigate two biomechanical properties of the cell, the intracellular water content and the hydraulic permeability. Intracellular water content did not vary between treatment groups, but hydraulic permeability increased significantly with inflammatory treatment. In the 1 week recovery group, hydraulic permeability remained elevated relative to the untreated recovery control. Cell radius was also significantly increased both after 24 hours of treatment and after 1 week recovery. A significant linear correlation was observed between hydraulic permeability and cell radius in untreated cells at 24 hours and at 1-week recovery, though not in the inflammatory stimulated groups at either time point. This loss of correlation between cell size and hydraulic permeability suggests that regulation of volume change is disrupted irreversibly due to inflammatory stimulation. Inflammatory treated cells exhibited altered F

  20. PRKAR1A inactivation leads to increased proliferation and decreased apoptosis in human B lymphocytes.

    PubMed

    Robinson-White, Audrey J; Leitner, Wolfgang W; Aleem, Eiman; Kaldis, Philipp; Bossis, Ioannis; Stratakis, Constantine A

    2006-11-01

    The multiple neoplasia syndrome Carney complex (CNC) is caused by heterozygote mutations in the gene, which codes for the RIalpha regulatory subunit (PRKAR1A) of protein kinase A. Inactivation of PRKAR1A and the additional loss of the normal allele lead to tumors in CNC patients and increased cyclic AMP signaling in their cells, but the oncogenetic mechanisms in affected tissues remain unknown. Previous studies suggested that PRKAR1A down-regulation may lead to increased mitogen-activated protein kinase (MAPK) signaling. Here, we show that, in lymphocytes with PRKAR1A-inactivating mutations, there is increased extracellular signal-regulated kinase (ERK) 1/2 and B-raf phosphorylation and MAPK/ERK kinase 1/2 and c-Myc activation, whereas c-Raf-1 is inhibited. These changes are accompanied by increased cell cycle rates and decreased apoptosis that result in an overall net gain in proliferation and survival. In conclusion, inactivation of PRKAR1A leads to widespread changes in molecular pathways that control cell cycle and apoptosis. This is the first study to show that human cells with partially inactivated RIalpha levels have increased proliferation and survival, suggesting that loss of the normal allele in these cells is not necessary for these changes to occur. PMID:17079485

  1. Inactivation of F-specific bacteriophages during flocculation with polyaluminum chloride - a mechanistic study.

    PubMed

    Kreißel, Katja; Bösl, Monika; Hügler, Michael; Lipp, Pia; Franzreb, Matthias; Hambsch, Beate

    2014-03-15

    Bacteriophages are often used as surrogates for enteric viruses in spiking experiments to determine the efficiencies of virus removal of certain water treatment measures, like e.g. flocculation or filtration steps. Such spiking experiments with bacteriophages are indispensable if the natural virus concentrations in the raw water of water treatment plants are too low to allow the determination of elimination levels over several orders of magnitude. In order to obtain reliable results from such spiking tests, it is essential that bacteriophages behave comparable to viruses and remain stable during the experiments. To test this, the influence of flocculation parameters on the bacteriophages MS2, Qβ and phiX174 was examined. Notably, the F-specific phages MS2 and Qβ were found to be inactivated in flocculation processes with polyaluminum chloride (PACl). In contrast, other aluminum coagulants like AlCl3 or Al2(SO4)3 did not show a comparable effect on MS2 in this study. In experiments testing the influence of different PACl species on MS2 and Qβ inactivation during flocculation, it could be shown that cationic dissolved PACl species (Al13) interacted with the MS2 surface and hereby reduced the surviving phage fraction to c/c0 values below 1*10(-4) even at very low PACl concentrations of 7 μmol Al/L. Other inactivation mechanisms like the irreversible adsorption of phages to the floc structure or the damage of phage surfaces due to entrapment into the floc during coagulation and floc formation do not seem to contribute to the low surviving fraction found for both F-specific bacteriophages. Furthermore, no influence of phage agglomeration or pH drops during the flocculation process on phage inactivation could be observed. The somatic coliphage phiX174 in contrast did not show sensitivity to chemical stress and in accordance only slight interaction between Al13 and the phage surface was observed. Consequently, F-specific phages like MS2 should not be used as

  2. Inactivation of human interferon by body fluids

    NASA Technical Reports Server (NTRS)

    Cesario, T. C.; Mandell, A.; Tilles, J. G.

    1973-01-01

    Description of the effects of human feces, bile, saliva, serum, and cerebrospinal fluid on interferon activity. It is shown that crude interferon is inactivated by at least 50% more than with the control medium used, when incubated for 4 hr in vitro in the presence of serum, saliva, or cerebrospinal liquid, and by close to 100% when incubated with stool extract or bile.

  3. Inactivation of Bacillus atrophaeus by OH radicals

    NASA Astrophysics Data System (ADS)

    Ono, Ryo; Yonetamari, Kenta; Tokumitsu, Yusuke; Yonemori, Seiya; Yasuda, Hachiro; Mizuno, Akira

    2016-08-01

    The inactivation of Bacillus atrophaeus by OH radicals is measured. This study aims to evaluate the bactericidal effects of OH radicals produced by atmospheric-pressure nonthermal plasma widely used for plasma medicine; however, in this study, OH radicals are produced by vacuum ultraviolet (VUV) photolysis of water vapor instead of plasma to allow the production of OH radicals with almost no other reactive species. A 172 nm VUV light from a Xe2 excimer lamp irradiates a He–H2O mixture flowing in a quartz tube to photodissociate H2O to produce OH, H, O, HO2, H2O2, and O3. The produced reactive oxygen species (ROS) flow out of the quartz tube nozzle to the bacteria on an agar plate and cause inactivation. The inactivation by OH radicals among the six ROS is observed by properly setting the experimental conditions with the help of simulations calculating the ROS densities. A 30 s treatment with approximately 0.1 ppm OH radicals causes visible inactivation.

  4. Temperature Tolerance and Inactivation of Chikungunya Virus.

    PubMed

    Huang, Yan-Jang S; Hsu, Wei-Wen; Higgs, Stephen; Vanlandingham, Dana L

    2015-11-01

    In late 2013, chikungunya virus (CHIKV) was introduced to the New World and large outbreaks occurred in the Caribbean islands causing over a million suspected and over 20,000 laboratory-confirmed cases. Serological analysis is an essential component for the diagnosis of CHIKV infection together with virus isolation and detection of viral nucleic acid. Demonstrating virus neutralizing by serum antibodies in a plaque reduction neutralization test (PRNT) is the gold standard of all serological diagnostic assays. Prior to the testing, heat inactivation of serum at 56°C for 30 min is required for the inactivation of complement activity and adventitious viruses. The presence of adventitious contaminating viruses may interfere with the results by leading to a higher number of plaques on the monolayers and subsequent false-negative results. This procedure is widely accepted for the inactivation of flaviviruses and alphaviruses. In this study, the thermostability of CHIKV was evaluated. Heat inactivation at 56°C for 30 min was demonstrated to be insufficient for the complete removal of infectious CHIKV virions present in the samples. This thermotolerance of CHIKV could compromise the accuracy of serum tests, and therefore longer treatment for greater than 120 min is recommended.

  5. Inactivation of human norovirus using chemical sanitizers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The porcine gastric mucin binding magnetic bead (PGM-MB) assay was used to evaluate the ability of chlorine, chlorine dioxide, peroxyacetic acid, hydrogen peroxide, and trisodium phosphate to inactivate human norovirus within 10 percent stool filtrate. One min free chlorine treatments at concentrat...

  6. Inactivation of Bacillus atrophaeus by OH radicals

    NASA Astrophysics Data System (ADS)

    Ono, Ryo; Yonetamari, Kenta; Tokumitsu, Yusuke; Yonemori, Seiya; Yasuda, Hachiro; Mizuno, Akira

    2016-08-01

    The inactivation of Bacillus atrophaeus by OH radicals is measured. This study aims to evaluate the bactericidal effects of OH radicals produced by atmospheric-pressure nonthermal plasma widely used for plasma medicine; however, in this study, OH radicals are produced by vacuum ultraviolet (VUV) photolysis of water vapor instead of plasma to allow the production of OH radicals with almost no other reactive species. A 172 nm VUV light from a Xe2 excimer lamp irradiates a He-H2O mixture flowing in a quartz tube to photodissociate H2O to produce OH, H, O, HO2, H2O2, and O3. The produced reactive oxygen species (ROS) flow out of the quartz tube nozzle to the bacteria on an agar plate and cause inactivation. The inactivation by OH radicals among the six ROS is observed by properly setting the experimental conditions with the help of simulations calculating the ROS densities. A 30 s treatment with approximately 0.1 ppm OH radicals causes visible inactivation.

  7. Monochloramine inactivation of bacterial select agents.

    PubMed

    Rose, Laura J; Rice, Eugene W; Hodges, Lisa; Peterson, Alicia; Arduino, Matthew J

    2007-05-01

    Seven species of bacterial select agents were tested for susceptibility to monochloramine. Under test conditions, the monochloramine routinely maintained in potable water would reduce six of the species by 2 orders of magnitude within 4.2 h. Bacillus anthracis spores would require up to 3.5 days for the same inactivation with monochloramine.

  8. High Pressure Inactivation of HAV within Mussels

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The potential of hepatitis A virus (HAV) to be inactivated within Mediterranean mussels (Mytilus galloprovincialis) and blue mussels (Mytilus edulis) by high pressure processing was evaluated. HAV was bioaccumulated within mussels to approximately 6-log10 PFU by exposure of mussels to HAV-contamina...

  9. Irreversible entropy production in two-phase flows with evaporating drops

    NASA Technical Reports Server (NTRS)

    Bellan, J.; Okong'o, N. A.

    2002-01-01

    A derivation of the irreversible entropy production, that is the dissipation, in two-phase flows is presented for the purpose of examining the effect of evaporative-drop modulation of flows having turbulent features.

  10. The Effects of Internal and External Irreversibility of a Vapor Compression Refrigeration Cycle

    NASA Astrophysics Data System (ADS)

    Wang, Fu-Jen; Chiou, Jeng-Shing

    The concept of finite-time thermodynamics is employed to investigate the optimal refrigeration rate for an irreversible refrigeration cycle. The heat transfer between the system (internal) fluid and cooling (external) fluid takes place at the actual heat exchanger, which has the finite-size heat transfer area and the realistic heat transfer effectiveness. The internal irreversibility results from the compression process and the expansion process are also considered. The optimal refrigeration rate is calculated and expressed in terms of the irreversibility parameter (Ir), coefficient of performance (COP), the time ratio(γ) of heat transfer processes and the effectiveness of heat exchanger. The derived COP which consider both the external and internal irreversibility can thus be considered as the benchmark value for a practical refrigeration cycle, and the parametric study can provide the basis for both determination of optimal operating conditions and design of a practical refrigeration cycle.

  11. Inactivation of malonate semialdehyde decarboxylase by 3-halopropiolates: evidence for hydratase activity.

    PubMed

    Poelarends, Gerrit J; Serrano, Hector; Johnson, William H; Whitman, Christian P

    2005-07-01

    Malonate semialdehyde decarboxylase (MSAD) from Pseudomonas pavonaceae 170 catalyzes the metal ion-independent decarboxylation of malonate semialdehyde and represents one of three known enzymatic activities in the tautomerase superfamily. The characterized members of this superfamily are structurally homologous proteins that share a beta-alpha-beta fold and a catalytic amino-terminal proline. Sequence analysis, chemical labeling studies, site-directed mutagenesis, and NMR studies of MSAD identified Pro-1 as a key active site residue in which the amino group has a pKa value of 9.2. The available evidence suggests a mechanism involving polarization of the C-3 carbonyl group of malonate semialdehyde by the cationic Pro-1. A second critical active site residue, Arg-75, could assist in the reaction by placing the substrate's carboxylate group in a favorable conformation for decarboxylation. In addition to the decarboxylase activity, MSAD has a hydratase activity as demonstrated by the MSAD-catalyzed conversion of 2-oxo-3-pentynoate to acetopyruvate. In view of this activity, MSAD was incubated with 3-bromo- and 3-chloropropiolate, and the subsequent reactions were characterized. Both compounds result in the irreversible inactivation of MSAD, making them the first identified inhibitors of MSAD. Inactivation by 3-chloropropiolate occurs in a time- and concentration-dependent manner and is due to the covalent modification of Pro-1. The proposed mechanism for inactivation involves the initial hydration of the 3-halopropiolate followed by a rearrangement to an alkylating agent, either an acyl halide or a ketene. The results provide additional evidence for the hydratase activity of MSAD and further support for the hypothesis that MSAD and trans-3-chloroacrylic acid dehalogenase, the preceding enzyme in the trans-1,3-dichloropropene catabolic pathway, diverged from a common ancestor but conserved the necessary catalytic machinery for the conjugate addition of water.

  12. Mechanisms of fusicoccin action: kinetic modification and inactivation of K(+) channels in guard cells.

    PubMed

    Blatt, M R; Clint, G M

    1989-12-01

    Fusicoccin commonly is thought to promote secondary solute transport via an increase in electrical driving force which follows the enhancement of primary, "electrogenic" H(+) extrusion by the plant plasma membrane H(+)-ATPase. However, previous electrical studies ofVicia faba L. guard cells in FC (Blatt, 1988, Planta174, 187) demonstrated, in addition to a limited rise in pump current, appreciable declines in membrane conductance near and positive to the free-running membrane potential (V m). Much of the current at these potentials could have been carried by outward-rectifying K(+) channels which were progressively inactivated in FC. We have examined this possibility in electrical studies, using whole-cell currents measured under voltage clamp to quantitate steadystate and kinetic characteristics of the K(+) channels both before and during exposure to FC; channels block in tetraethylammonium chloride was exploited to assess changes in background 'leak' currents. The cells showed little evidence of primary pump activity, a fact which further simplified analyses. Under these conditions, outward-directed K(+) channel current contributed to charge balance maintainingV m, and adding 10 μM FC on average depolarized (positive-going)V m. Steady-state current-voltage relations revealed changes both in K(+) channel and in leak currents underlying the voltage response. Changes in the leak were variable, but on average the leak equilibrium potential was shifted (+)19 mV and leak conductance declined by 21% over 30-40 min in FC. Potassium currents were inactivated irreversibly and with halftimes (current maxima) of 6.2-10.7 min. Inactivation was voltage-dependent, so that the activation ("gating") potential for the current was shifted, positive-going, with time in FC. Channel gating kinetics, inferred from the macroscopic currents, were also affected; current rise at positive potentials accelerated 4.5-fold and more, but in a manner apparently independent of voltage and

  13. Functional inactivation of Rb sensitizes cancer cells to TSC2 inactivation induced cell Death

    PubMed Central

    Danos, Arpad M.; Liao, Yang; Li, Xuan; Du, Wei

    2012-01-01

    We showed previously that inactivation of TSC2 induces death in cancer cells lacking the Retinoblastoma (Rb) tumor suppressor under stress conditions, suggesting that inactivation of TSC2 can potentially be used as an approach to specifically kill cancers that have lost WT Rb. As Rb is often inactivated in cancers by overexpression of cyclin D1, loss of p16ink4a cdk inhibitor, or expression of viral oncoproteins, it will be interesting to determine if such functional inactivation of Rb would similarly sensitize cancer cells to TSC2 inactivation induced cell death. In addition, many cancers lack functional Pten, resulting in increased PI3K/Akt signaling that has been shown to modulate E2F-induced cell death. Therefore it will be interesting to test whether loss of Pten will affect TSC2 inactivation induced killing of Rb mutant cancer cells. Here, we show that overexpression of Cyclin D1 or the viral oncogene E1a sensitizes cancer cells to TSC2 knockdown induced cell death and growth inhibition. On the other hand, knockdown of p16ink4a sensitizes cancer cells to TSC2 knockdown induced cell death in a manner that is likely dependant on serum induction of Cyclin D1 to inactivate the Rb function. Additionally, we demonstrate that loss of Pten does not interfere with TSC2 knockdown induced cell death in Rb mutant cancer cells. Together, these results suggest that TSC2 is potentially a useful target for a large spectrum of cancer types with an inactivated Rb pathway. PMID:23022476

  14. Photo-inactivation of Bacillus endospores: inter-specific variability of inactivation efficiency.

    PubMed

    da Silva, Raquel N; Tomé, Augusto C; Tomé, João P C; Neves, Maria G P M S; Faustino, Maria A F; Cavaleiro, José A S; Oliveira, Anabela; Almeida, Adelaide; Cunha, Ângela

    2012-10-01

    The aims of this work were to (a) evaluate the susceptibility of endospores of Bacillus cereus, B. licheniformis, B. sphaericus and B. subtilis to photodynamic inactivation using a tricationic porphyrin as photosensitizer, (b) assess the efficiency of adsorption of the photosensitizer in endospore material as a determinant of the susceptibility of endospores of different Bacillus species to photo-inactivation, (c) determine the value of B. cereus as a model organism for studies of antimicrobial photodynamic inactivation of bacterial endospores. The results of irradiation experiments with endospores of four species of Bacillus showed that B. cereus was the only species for which efficient endospore photo-inactivation (> 3 log reduction) could be achieved. Endospores of B. licheniformis, B. sphaericus and B. subtilis were virtually resistant to photo-inactivation with tricationic porphyrin. The amount of porphyrin bound to endospore material was not significantly different between species, regardless of the presence of an exosporium or exosporium-like outer layer. The sensitivity of endospores to photodynamic inactivation with a tricationic porphyrin is highly variable among different species of the genus Bacillus. The presence of an exosporium in endospores of B. cereus and B. sphaericus, or an exosporium-like glycoprotein layer in endospores of B. subtilis, did not affect the amount of bound photosensitizer and did not explain the inter-species variability in susceptibility to photodynamic inactivation. The results imply that the use of B. cereus as a more amenable surrogate of the exosporium-producing B. anthracis must be carefully considered when testing new photosensitizers for their antimicrobial photo-inactivation properties.

  15. Timescales and Frequencies of Reversible and Irreversible Adhesion Events of Single Bacterial Cells.

    PubMed

    Hoffman, Michelle D; Zucker, Lauren I; Brown, Pamela J B; Kysela, David T; Brun, Yves V; Jacobson, Stephen C

    2015-12-15

    In the environment, most bacteria form surface-attached cell communities called biofilms. The attachment of single cells to surfaces involves an initial reversible stage typically mediated by surface structures such as flagella and pili, followed by a permanent adhesion stage usually mediated by polysaccharide adhesives. Here, we determine the absolute and relative timescales and frequencies of reversible and irreversible adhesion of single cells of the bacterium Caulobacter crescentus to a glass surface in a microfluidic device. We used fluorescence microscopy of C. crescentus expressing green fluorescent protein to track the swimming behavior of individual cells prior to adhesion, monitor the cell at the surface, and determine whether the cell reversibly or irreversibly adhered to the surface. A fluorescently labeled lectin that binds specifically to polar polysaccharides, termed holdfast, discriminated irreversible adhesion events from reversible adhesion events where no holdfast formed. In wild-type cells, the holdfast production time for irreversible adhesion events initiated by surface contact (23 s) was 30-times faster than the holdfast production time that occurs through developmental regulation (13 min). Irreversible adhesion events in wild-type cells (3.3 events/min) are 15-times more frequent than in pilus-minus mutant cells (0.2 events/min), indicating the pili are critical structures in the transition from reversible to irreversible surface-stimulated adhesion. In reversible adhesion events, the dwell time of cells at the surface before departing was the same for wild-type cells (12 s) and pilus-minus mutant cells (13 s), suggesting the pili do not play a significant role in reversible adhesion. Moreover, reversible adhesion events in wild-type cells (6.8 events/min) occur twice as frequently as irreversible adhesion events (3.3 events/min), demonstrating that most cells contact the surface multiple times before transitioning from reversible to

  16. Timescales and Frequencies of Reversible and Irreversible Adhesion Events of Single Bacterial Cells.

    PubMed

    Hoffman, Michelle D; Zucker, Lauren I; Brown, Pamela J B; Kysela, David T; Brun, Yves V; Jacobson, Stephen C

    2015-12-15

    In the environment, most bacteria form surface-attached cell communities called biofilms. The attachment of single cells to surfaces involves an initial reversible stage typically mediated by surface structures such as flagella and pili, followed by a permanent adhesion stage usually mediated by polysaccharide adhesives. Here, we determine the absolute and relative timescales and frequencies of reversible and irreversible adhesion of single cells of the bacterium Caulobacter crescentus to a glass surface in a microfluidic device. We used fluorescence microscopy of C. crescentus expressing green fluorescent protein to track the swimming behavior of individual cells prior to adhesion, monitor the cell at the surface, and determine whether the cell reversibly or irreversibly adhered to the surface. A fluorescently labeled lectin that binds specifically to polar polysaccharides, termed holdfast, discriminated irreversible adhesion events from reversible adhesion events where no holdfast formed. In wild-type cells, the holdfast production time for irreversible adhesion events initiated by surface contact (23 s) was 30-times faster than the holdfast production time that occurs through developmental regulation (13 min). Irreversible adhesion events in wild-type cells (3.3 events/min) are 15-times more frequent than in pilus-minus mutant cells (0.2 events/min), indicating the pili are critical structures in the transition from reversible to irreversible surface-stimulated adhesion. In reversible adhesion events, the dwell time of cells at the surface before departing was the same for wild-type cells (12 s) and pilus-minus mutant cells (13 s), suggesting the pili do not play a significant role in reversible adhesion. Moreover, reversible adhesion events in wild-type cells (6.8 events/min) occur twice as frequently as irreversible adhesion events (3.3 events/min), demonstrating that most cells contact the surface multiple times before transitioning from reversible to

  17. The Transient Inactivation of the Master Cell Cycle Phosphatase Cdc14 Causes Genomic Instability in Diploid Cells of Saccharomyces cerevisiae

    PubMed Central

    Quevedo, Oliver; Ramos-Pérez, Cristina; Petes, Thomas D.; Machín, Félix

    2015-01-01

    Genomic instability is a common feature found in cancer cells . Accordingly, many tumor suppressor genes identified in familiar cancer syndromes are involved in the maintenance of the stability of the genome during every cell division and are commonly referred to as caretakers. Inactivating mutations and epigenetic silencing of caretakers are thought to be the most important mechanisms that explain cancer-related genome instability. However, little is known of whether transient inactivation of caretaker proteins could trigger genome instability and, if so, what types of instability would occur. In this work, we show that a brief and reversible inactivation, during just one cell cycle, of the key phosphatase Cdc14 in the model organism Saccharomyces cerevisiae is enough to result in diploid cells with multiple gross chromosomal rearrangements and changes in ploidy. Interestingly, we observed that such transient loss yields a characteristic fingerprint whereby trisomies are often found in small-sized chromosomes, and gross chromosome rearrangements, often associated with concomitant loss of heterozygosity, are detected mainly on the ribosomal DNA-bearing chromosome XII. Taking into account the key role of Cdc14 in preventing anaphase bridges, resetting replication origins, and controlling spindle dynamics in a well-defined window within anaphase, we speculate that the transient loss of Cdc14 activity causes cells to go through a single mitotic catastrophe with irreversible consequences for the genome stability of the progeny. PMID:25971663

  18. The antibrowning agent sulfite inactivates Agaricus bisporus tyrosinase through covalent modification of the copper-B site.

    PubMed

    Kuijpers, Tomas F M; Gruppen, Harry; Sforza, Stefano; van Berkel, Willem J H; Vincken, Jean-Paul

    2013-12-01

    Sulfite salts are widely used as antibrowning agents in food processing. Nevertheless, the exact mechanism by which sulfite prevents enzymatic browning has remained unknown. Here, we show that sodium hydrogen sulfite (NaHSO3) irreversibly blocks the active site of tyrosinase from the edible mushroom Agaricus bisporus, and that the competitive inhibitors tropolone and kojic acid protect the enzyme from NaHSO3 inactivation. LC-MS analysis of pepsin digests of NaHSO3 -treated tyrosinase revealed two peptides showing a neutral loss corresponding to the mass of SO3 upon MS(2) fragmentation. These peptides were found to be homologous peptides containing two of the three histidine residues that form the copper-B-binding site of mushroom tyrosinase isoform PPO3 and mushroom tyrosinase isoform PPO4, which were both present in the tyrosinase preparation used. Peptides showing this neutral loss behavior were not found in the untreated control. Comparison of the effects of NaHSO3 on apo-tyrosinase and holo-tyrosinase indicated that inactivation is facilitated by the active site copper ions. These data provide compelling evidence that inactivation of mushroom tyrosinase by NaHSO3 occurs through covalent modification of a single amino-acid residue, probably via addition of HSO3(-) to one of the copper-coordinating histidines in the copper-B site of the enzyme.

  19. The antibrowning agent sulfite inactivates Agaricus bisporus tyrosinase through covalent modification of the copper-B site.

    PubMed

    Kuijpers, Tomas F M; Gruppen, Harry; Sforza, Stefano; van Berkel, Willem J H; Vincken, Jean-Paul

    2013-12-01

    Sulfite salts are widely used as antibrowning agents in food processing. Nevertheless, the exact mechanism by which sulfite prevents enzymatic browning has remained unknown. Here, we show that sodium hydrogen sulfite (NaHSO3) irreversibly blocks the active site of tyrosinase from the edible mushroom Agaricus bisporus, and that the competitive inhibitors tropolone and kojic acid protect the enzyme from NaHSO3 inactivation. LC-MS analysis of pepsin digests of NaHSO3 -treated tyrosinase revealed two peptides showing a neutral loss corresponding to the mass of SO3 upon MS(2) fragmentation. These peptides were found to be homologous peptides containing two of the three histidine residues that form the copper-B-binding site of mushroom tyrosinase isoform PPO3 and mushroom tyrosinase isoform PPO4, which were both present in the tyrosinase preparation used. Peptides showing this neutral loss behavior were not found in the untreated control. Comparison of the effects of NaHSO3 on apo-tyrosinase and holo-tyrosinase indicated that inactivation is facilitated by the active site copper ions. These data provide compelling evidence that inactivation of mushroom tyrosinase by NaHSO3 occurs through covalent modification of a single amino-acid residue, probably via addition of HSO3(-) to one of the copper-coordinating histidines in the copper-B site of the enzyme. PMID:24112416

  20. Influence of different types of effectors on the kinetic parameters of suicide inactivation of catalase by hydrogen peroxide.

    PubMed

    Ghadermarzi, M; Moosavi-Movahedi, A A

    1999-04-12

    The effects of cyanide and azide ions (class A), sodium-n-dodecyl sulphate (SDS) and 2-mercaptoethanol (class B), 3-aminotriazole (class C) and NADPH (class D) on the initial activity (ai), inactivation rate constant (ki) and the partition ratio (r) of bovine liver catalase reaction with its suicide substrate, hydrogen peroxide, were studied in 50 mM sodium phosphate buffer, pH 7.0 at 27 degrees C. The above kinetic parameters were determined by processing the progress curve data. In class A, which contains fast and reversible inhibitors of catalase, a proportional decrease in ai and ki was observed by inhibitors, so that the r remained constant. In class B, which contains slow and irreversible inactivators, a decrease in ai and constancy of ki and r were observed when catalase was incubated in the presence of such inactivators for a determined time. In class C, containing effector which can combine with intermediate compound I, ai was relatively unchanged but an increase in ki and a decrease in r were observed. In class D, containing effector which reduces compound I to ferricatalase, ai was not affected significantly but some decrease in ki was detected which was linked with an increase in r. These results demonstrate that different classes of effectors affect the determined kinetic parameters of catalase in various ways. Thus, determination of such parameters by simple kinetic experiments can be carried out for classification of the agents which have an effect on the kinetics of catalase. PMID:10209276

  1. Inactivation of Escherichia coli phage by pulsed electric field treatment and analysis of inactivation mechanism

    NASA Astrophysics Data System (ADS)

    Tanino, Takanori; Yoshida, Tomoki; Sakai, Kazuki; Ohshima, Takayuki

    2013-03-01

    Inactivation of bacteriophage by pulsed electric field (PEF) treatment, one of the effective procedures for bacteria nonthermal inactivation, was studied. Model phage particles Escherichia coli bacteriophages M13mp18 and λ phage, were successfully inactivated by PEF treatment. The survival ratios of both bacteriophages decreased depending on the PEF treatment time when applied peak voltage was 5 or 7 kV, and the survival ratios after 12 min PEF treatment were 10-4 - 10-5. Electrophoresis analyses of biological molecules of inactivated λ phage detected no degradation of total protein and genomic DNA. These results suggested that the factor of phage inactivation by PEF treatment was not based on the degradation of protein or DNA, but on the destruction of phage particle structure. Sensitivity of E. coli phage to PEF treatment was compared with that of E. coli cell. Phage and MV1184 cell were treated with same condition PEF at 5 kV, respectively. After 12 min treatment, the survival ration of λ phage and MV1184 were 4.0 × 10-5 and 1.7 × 10-3, respectively. The survival ratio of phage was lower than that of MV1184. E. coli cell is more tolerant to inactivation with PEF treatment than coli phage.

  2. Substance P and CGRP expression in dental pulps with irreversible pulpitis.

    PubMed

    Sattari, Mandana; Mozayeni, Mohammad Ali; Matloob, Arash; Mozayeni, Maryam; Javaheri, Homan H

    2010-08-01

    The purpose of this study was to compare substance P (SP) and calcitonin gene-related peptide (CGRP) expression in pulp tissue with clinically diagnosed symptomatic and asymptomatic irreversible pulpitis. Healthy pulps acted as controls. Five normal pulps and 40 with irreversible pulpitis (20 symptomatic and 20 asymptomatic) were obtained from 45 different patients. SP and CGRP expression was determined by competition binding assays using enzyme immunoassay. anova and Mann-Whitney tests were used to ascertain if there were statistically significant differences between the groups. The results showed that neuropeptides were found in all pulp samples. The highest and the lowest expressions for SP and CGRP were found in symptomatic irreversible pulpitis and healthy pulps groups, respectively. The differences between healthy pulps and the groups of pulps having irreversible pulpitis were significant (P < 0.001). Although Mann-Whitney's post-hoc tests showed statistically significant differences in CGRP expression between two pulpitis groups (P < 0.05), differences in SP expression between symptomatic and asymptomatic irreversible pulpitis groups were not significant. This study demonstrated that the expression of CGRP and SP is significantly higher in pulps with irreversible pulpitis compared with healthy pulps. PMID:20666750

  3. Inactivating effects of lignin-derived compounds released during lignocellulosic biomass pretreatment on the endo-glucanase catalyzed hydrolysis of carboxymethylcellulose: A study in continuous stirred ultrafiltration-membrane reactor.

    PubMed

    Cantarella, Maria; Mucciante, Claudia; Cantarella, Laura

    2014-03-01

    This study focusses on the reversible/irreversible damage that selected phenolic compounds, released during steam-explosion pretreatment, mandatory for cellulose accessibility, causes on both stability and activity of a commercial cellulase (half-life=173h) during carboxymethyl-cellulose hydrolysis. Long-term experiments performed in continuous stirred UF-membrane bioreactors, operating at steady-state regime, in controlled operational conditions, allowed evaluating the inactivation-constant in the phenol presence (kd1) and after its removal (kd2) from the reactor feed. p-Hydroxybenzoic acid (1 and 2g L(-1)) are the extreme limits in the inactivating effect with enzyme half-lives 99.02 and 14.15h, respectively. The inactivation reversibility was assessed for vanillic acid, p-hydroxybenzoic acid, syringaldehyde, p-coumaric acid, being kd1>kd2. p-Hydroxybenzaldehyde and protocatechuic acid irreversibly affected cellulase stability increasing its inactivation with kd2>kd1. p-Hydroxybenzaldehyde, 1g L(-1), syringaldehyde, and vanillin, at 2gL(-1), had similar kd1÷kd2. PMID:24486937

  4. Inactivating effects of lignin-derived compounds released during lignocellulosic biomass pretreatment on the endo-glucanase catalyzed hydrolysis of carboxymethylcellulose: A study in continuous stirred ultrafiltration-membrane reactor.

    PubMed

    Cantarella, Maria; Mucciante, Claudia; Cantarella, Laura

    2014-03-01

    This study focusses on the reversible/irreversible damage that selected phenolic compounds, released during steam-explosion pretreatment, mandatory for cellulose accessibility, causes on both stability and activity of a commercial cellulase (half-life=173h) during carboxymethyl-cellulose hydrolysis. Long-term experiments performed in continuous stirred UF-membrane bioreactors, operating at steady-state regime, in controlled operational conditions, allowed evaluating the inactivation-constant in the phenol presence (kd1) and after its removal (kd2) from the reactor feed. p-Hydroxybenzoic acid (1 and 2g L(-1)) are the extreme limits in the inactivating effect with enzyme half-lives 99.02 and 14.15h, respectively. The inactivation reversibility was assessed for vanillic acid, p-hydroxybenzoic acid, syringaldehyde, p-coumaric acid, being kd1>kd2. p-Hydroxybenzaldehyde and protocatechuic acid irreversibly affected cellulase stability increasing its inactivation with kd2>kd1. p-Hydroxybenzaldehyde, 1g L(-1), syringaldehyde, and vanillin, at 2gL(-1), had similar kd1÷kd2.

  5. Effects of Bacterial Inactivation Methods on Downstream Proteomic Analysis

    SciTech Connect

    Lin, Andy; Merkley, Eric D.; Clowers, Brian H.; Hutchison, Janine R.; Kreuzer, Helen W.

    2015-05-01

    Inactivation of pathogenic microbial samples is often necessary for the protection of researchers and to comply with local and federal regulations. By its nature, biological inactivation causes changes to microbial samples, potentially affecting observed experimental results. While inactivation induced damage to materials such as DNA has been evaluated, the effect of various inactivation strategies on proteomic data, to our knowledge, has not been discussed. To this end, we inactivated samples of Yersinia pestis and Escherichia coli by autoclave, ethanol, or irradiation treatment to determine how inactivation changes liquid chromatography tandem mass spectrometry data quality as well as apparent protein content of cells. Proteomic datasets obtained from aliquots of samples inactivated by different methods were highly similar, with Pearson correlation coefficients ranging from 0.822 to 0.985 and 0.816 to 0.985 for E. coli and Y. pestis, respectively, suggesting that inactivation had only slight impacts on the set of proteins identified. In addition, spectral quality metrics such as distributions of various database search algorithm scores remained constant across inactivation methods, indicating that inactivation does not appreciably degrade spectral quality. Though overall changes resulting from inactivation were small, there were detectable trends. For example, one-sided Fischer exact tests determined that periplasmic proteins decrease in observed abundance after sample inactivation by autoclaving (α = 1.71x10-2 for E. coli, α = 4.97x10-4 for Y. pestis) and irradiation (α = 9.43x10-7 for E. coli, α = 1.21x10-5 for Y. pestis) when compared to controls that were not inactivated. Based on our data, if sample inactivation is necessary, we recommend inactivation with ethanol treatment with secondary preference given to irradiation.

  6. Effects of bacterial inactivation methods on downstream proteomic analysis.

    PubMed

    Lin, Andy; Merkley, Eric D; Clowers, Brian H; Hutchison, Janine R; Kreuzer, Helen W

    2015-05-01

    Inactivation of pathogenic microbial samples is often necessary for the protection of researchers and to comply with local and federal regulations. By its nature, biological inactivation causes changes to microbial samples, potentially affecting observed experimental results. While inactivation-induced damage to materials such as DNA has been evaluated, the effect of various inactivation strategies on proteomic data, to our knowledge, has not been discussed. To this end, we inactivated samples of Yersinia pestis and Escherichia coli by autoclave, ethanol, or irradiation treatment to determine how inactivation changes liquid chromatography-tandem mass spectrometry data quality as well as apparent protein content of cells. Proteomic datasets obtained from aliquots of samples inactivated by different methods were highly similar, with Pearson correlation coefficients ranging from 0.822 to 0.985 and 0.816 to 0.985 for E. coli and Y. pestis, respectively, suggesting that inactivation had only slight impacts on the set of proteins identified. In addition, spectral quality metrics such as distributions of various database search algorithm scores remained constant across inactivation methods, indicating that inactivation does not appreciably degrade spectral quality. Though overall changes resulting from inactivation were small, there were detectable trends. For example, one-sided Fischer exact tests determined that periplasmic proteins decrease in observed abundance after sample inactivation by autoclaving (α=1.71×10(-2) for E. coli, α=4.97×10(-4) for Y. pestis) and irradiation (α=9.43×10(-7) for E. coli, α=1.21×10(-5) for Y. pestis) when compared to controls that were not inactivated. Based on our data, if sample inactivation is necessary, we recommend inactivation with ethanol treatment with secondary preference given to irradiation. PMID:25620019

  7. Time-dependent inactivation of chick-embryo prolyl 4-hydroxylase by coumalic acid. Evidence for a syncatalytic mechanism.

    PubMed Central

    Günzler, V; Hanauske-Abel, H M; Myllylä, R; Mohr, J; Kivirikko, K I

    1987-01-01

    From the structure-activity relationships of known competitive inhibitors, coumalic acid (2-oxo-1,2H-pyran-5-carboxylic acid) was deduced to be a potential syncatalytic inhibitor for chick-embryo prolyl 4-hydroxylase. The compound caused time-dependent inactivation, the reaction rate being first-order. The inactivation constant was 0.094 min-1, the Ki 17 mM and the bimolecular rate constant 0.09 M-1 X S-1. Human prolyl 4-hydroxylase and chick embryo lysyl hydroxylase were also inactivated, though to a lesser extent. Inactivation could be prevented by adding high concentrations of 2-oxoglutarate or its competitive analogues to the reaction mixture. In Lineweaver-Burk kinetics, coumalic acid displayed S-parabolic competitive inhibition with respect to 2-oxoglutarate. The inactivation reaction had cofactor requirements similar to those for the decarboxylation of 2-oxoglutarate. Enzymic activity was partially preserved in the absence of iron, but the rescue was incomplete, owing to decreased stability of the enzyme under this condition. Coumalic acid also decreased the electrophoretic mobility of the alpha-subunit, but the beta-subunit was not affected. Prolonged incubation of coumalic acid above pH 6.8 led to loss of its inactivating potency, owing to hydrolysis. It is concluded that the inactivation of prolyl 4-hydroxylase by coumalic acid is due to a syncatalytic mechanism. The data also suggest that the 2-oxoglutarate-binding site of the enzyme is located within the alpha-subunit. PMID:3036081

  8. Irreversible denaturation of maltodextrin glucosidase studied by differential scanning calorimetry, circular dichroism, and turbidity measurements.

    PubMed

    Goyal, Megha; Chaudhuri, Tapan K; Kuwajima, Kunihiro

    2014-01-01

    Thermal denaturation of Escherichia coli maltodextrin glucosidase was studied by differential scanning calorimetry, circular dichroism (230 nm), and UV-absorption measurements (340 nm), which were respectively used to monitor heat absorption, conformational unfolding, and the production of solution turbidity. The denaturation was irreversible, and the thermal transition recorded at scan rates of 0.5-1.5 K/min was significantly scan-rate dependent, indicating that the thermal denaturation was kinetically controlled. The absence of a protein-concentration effect on the thermal transition indicated that the denaturation was rate-limited by a mono-molecular process. From the analysis of the calorimetric thermograms, a one-step irreversible model well represented the thermal denaturation of the protein. The calorimetrically observed thermal transitions showed excellent coincidence with the turbidity transitions monitored by UV-absorption as well as with the unfolding transitions monitored by circular dichroism. The thermal denaturation of the protein was thus rate-limited by conformational unfolding, which was followed by a rapid irreversible formation of aggregates that produced the solution turbidity. It is thus important to note that the absence of the protein-concentration effect on the irreversible thermal denaturation does not necessarily means the absence of protein aggregation itself. The turbidity measurements together with differential scanning calorimetry in the irreversible thermal denaturation of the protein provided a very effective approach for understanding the mechanisms of the irreversible denaturation. The Arrhenius-equation parameters obtained from analysis of the thermal denaturation were compared with those of other proteins that have been reported to show the one-step irreversible thermal denaturation. Maltodextrin glucosidase had sufficiently high kinetic stability with a half-life of 68 days at a physiological temperature (37°C).

  9. [Inactivation of concentrated biomasses of Rickettsia prowazekii].

    PubMed

    Eremeeva, M E; Ignatovich, V F; Popov, V L; Balaeva, N M

    1989-07-01

    The methods used for the sparing inactivation of highly concentrated R. prowazekii biomass and for the decrease of its infectious activity are described. These methods are recommended for use in experiments in the field of molecular biology, as well as for disinfection of different materials contaminated with rickettsiae. As conditions for complete inactivation, incubation at 50 degrees C for 1 hour without chemical disinfectants, treatment with 0.5% phenol solution at 30 degrees C for 12 hours and with 0.1% formaldehyde solution at 4 degrees C for 24 hours have been selected. Treatment with 0.5% phenol solution at 36 degrees C for 1 hour or incubation at 45 degrees C without the use of disinfectants ensures an essential decrease in the infectivity of the material if the work with viable infective agents is necessary. Ultraviolet irradiation for 1.5 hours and exposure to the action of 0.1-0.5% sodium azide are less effective.

  10. Thermal inactivation of Alkhumra hemorrhagic fever virus.

    PubMed

    Madani, Tariq A; Abuelzein, El-Tayb M E; Azhar, Esam I; Al-Bar, Hussein M S

    2014-10-01

    The physico-chemical and biological characteristics of Alkhumra hemorrhagic fever virus (AHFV) are not yet known. The present study describes the thermal stability of this virus at different temperatures for different periods. The kinetics of thermal inactivation were studied, linear regressions were plotted, the Arrhenius equation was applied, and the activation energy was calculated accordingly. Titers of the residual virus were determined in median tissue culture infective dose (TCID50), and the rate of destruction of infectivity at various temperatures was determined. Infectivity of AHFV was completely lost upon heating for 3 minutes at 60 °C and for 30 min at 56 °C. However, the virus could maintain 33.2 % of its titer after heating for 60 min at 45 °C and 32 % of its titer after heating for 60 min at 50 °C. In conclusion, AHFV is thermo-labile, and its inactivation follows first-order kinetics. PMID:24906524

  11. Ixodes dammini: salivary anaphylatoxin inactivating activity.

    PubMed

    Ribeiro, J M; Spielman, A

    1986-10-01

    Saliva of the tick, Ixodes dammini, antagonizes anaphylatoxin, abolishing both the effects of anaphylatoxin on guinea pig ileum preparations regularly stimulated with histamine and the local edema caused by intradermal injection of anaphylatoxin into guinea pigs. Saliva of these ticks, however, did not modify polymorphonuclear leukocyte aggregation induced by anaphylatoxin. Bradykinin and lysil-bradykinin were inactivated, but angiotensin I, angiotensin II, and substance P were not affected. Amino acids were released rapidly following incubation of saliva with bradykinin, but slowly from des-arg-9-bradykinin. These results suggest the presence of a salivary carboxypeptidase with specificity for terminal basic amino acids. Such activity may inactivate anaphylatoxin and bradykinin at the site of tick attachment. PMID:3743719

  12. Recurrent inactivating RASA2 mutations in melanoma

    PubMed Central

    Arafeh, Rand; Qutob, Nouar; Emmanuel, Rafi; Keren-Paz, Alona; Madore, Jason; Elkahloun, Abdel; Wilmott, James S.; Gartner, Jared J.; Di Pizio, Antonella; Winograd-Katz, Sabina; Sindiri, Sivasish; Rotkopf, Ron; Dutton-Regester, Ken; Johansson, Peter; Pritchard, Antonia; Waddell, Nicola; Hill, Victoria K.; Lin, Jimmy C.; Hevroni, Yael; Rosenberg, Steven A.; Khan, Javed; Ben-Dor, Shifra; Niv, Masha Y.; Ulitsky, Igor; Mann, Graham J; Scolyer, Richard A.; Hayward, Nicholas K.; Samuels, Yardena

    2016-01-01

    Analysis of 501 melanoma exomes revealed RASA2, encoding a RasGAP, as a tumor-suppressor gene mutated in 5% of melanomas. Recurrent loss-of-function mutations in RASA2 were found to increase RAS activation, melanoma cell growth and migration. RASA2 expression was lost in ≥30% of human melanomas and was associated with reduced patient survival. These findings reveal RASA2 inactivation as a melanoma driver and highlight the importance of Ras GAPs in cancer. PMID:26502337

  13. Female meiotic sex chromosome inactivation in chicken.

    PubMed

    Schoenmakers, Sam; Wassenaar, Evelyne; Hoogerbrugge, Jos W; Laven, Joop S E; Grootegoed, J Anton; Baarends, Willy M

    2009-05-01

    During meiotic prophase in male mammals, the heterologous X and Y chromosomes remain largely unsynapsed, and meiotic sex chromosome inactivation (MSCI) leads to formation of the transcriptionally silenced XY body. In birds, the heterogametic sex is female, carrying Z and W chromosomes (ZW), whereas males have the homogametic ZZ constitution. During chicken oogenesis, the heterologous ZW pair reaches a state of complete heterologous synapsis, and this might enable maintenance of transcription of Z- and W chromosomal genes during meiotic prophase. Herein, we show that the ZW pair is transiently silenced, from early pachytene to early diplotene using immunocytochemistry and gene expression analyses. We propose that ZW inactivation is most likely achieved via spreading of heterochromatin from the W on the Z chromosome. Also, persistent meiotic DNA double-strand breaks (DSBs) may contribute to silencing of Z. Surprisingly, gammaH2AX, a marker of DSBs, and also the earliest histone modification that is associated with XY body formation in mammalian and marsupial spermatocytes, does not cover the ZW during the synapsed stage. However, when the ZW pair starts to desynapse, a second wave of gammaH2AX accumulates on the unsynapsed regions of Z, which also show a reappearance of the DSB repair protein RAD51. This indicates that repair of meiotic DSBs on the heterologous part of Z is postponed until late pachytene/diplotene, possibly to avoid recombination with regions on the heterologously synapsed W chromosome. Two days after entering diplotene, the Z looses gammaH2AX and shows reactivation. This is the first report of meiotic sex chromosome inactivation in a species with female heterogamety, providing evidence that this mechanism is not specific to spermatogenesis. It also indicates the presence of an evolutionary force that drives meiotic sex chromosome inactivation independent of the final achievement of synapsis. PMID:19461881

  14. Female Meiotic Sex Chromosome Inactivation in Chicken

    PubMed Central

    Schoenmakers, Sam; Wassenaar, Evelyne; Hoogerbrugge, Jos W.; Laven, Joop S. E.; Grootegoed, J. Anton; Baarends, Willy M.

    2009-01-01

    During meiotic prophase in male mammals, the heterologous X and Y chromosomes remain largely unsynapsed, and meiotic sex chromosome inactivation (MSCI) leads to formation of the transcriptionally silenced XY body. In birds, the heterogametic sex is female, carrying Z and W chromosomes (ZW), whereas males have the homogametic ZZ constitution. During chicken oogenesis, the heterologous ZW pair reaches a state of complete heterologous synapsis, and this might enable maintenance of transcription of Z- and W chromosomal genes during meiotic prophase. Herein, we show that the ZW pair is transiently silenced, from early pachytene to early diplotene using immunocytochemistry and gene expression analyses. We propose that ZW inactivation is most likely achieved via spreading of heterochromatin from the W on the Z chromosome. Also, persistent meiotic DNA double-strand breaks (DSBs) may contribute to silencing of Z. Surprisingly, γH2AX, a marker of DSBs, and also the earliest histone modification that is associated with XY body formation in mammalian and marsupial spermatocytes, does not cover the ZW during the synapsed stage. However, when the ZW pair starts to desynapse, a second wave of γH2AX accumulates on the unsynapsed regions of Z, which also show a reappearance of the DSB repair protein RAD51. This indicates that repair of meiotic DSBs on the heterologous part of Z is postponed until late pachytene/diplotene, possibly to avoid recombination with regions on the heterologously synapsed W chromosome. Two days after entering diplotene, the Z looses γH2AX and shows reactivation. This is the first report of meiotic sex chromosome inactivation in a species with female heterogamety, providing evidence that this mechanism is not specific to spermatogenesis. It also indicates the presence of an evolutionary force that drives meiotic sex chromosome inactivation independent of the final achievement of synapsis. PMID:19461881

  15. Inactivation of Anandamide Signaling: A Continuing Debate

    PubMed Central

    Khairy, Hesham; Houssen, Wael E.

    2010-01-01

    Since the first endocannabinoid anandamide was identified in 1992, extensive research has been conducted to characterize the elements of the tightly controlled endocannabinoid signaling system. While it was established that the activity of endocannabinoids are terminated by a two-step process that includes cellular uptake and degradation, there is still a continuing debate about the mechanistic role of these processes in inactivating anandamide signals.

  16. Hyaluronan decreases surfactant inactivation in vitro.

    PubMed

    Lu, Karen W; Goerke, Jon; Clements, John A; Taeusch, H William

    2005-02-01

    Hyaluronan (HA) is an anionic polymer and a constituent of alveolar fluid that can bind proteins, phospholipids, and water. Previous studies have established that nonionic polymers improve the surface activity of pulmonary surfactants by decreasing inactivation of surfactant. In this work, we investigate whether HA can also have beneficial effects when added to surfactants. We used a modified pulsating bubble surfactometer to measure mixtures of several commercially available pulmonary surfactants or native calf surfactant with and without serum inactivation. Surface properties such as equilibrium surface tension, minimum and maximum surface tensions on compression and expansion of a surface film, and degree of surface area reduction required to reach a surface tension of 10 mN/m were measured. In the presence of serum, addition of HA dramatically improved the surface activities of all four surfactants and in some cases in the absence of serum as well. These results indicate that HA reduces inactivation of surfactants caused by serum and add evidence that endogenous HAs may interact with alveolar surfactant under normal and abnormal conditions.

  17. Rapid inactivation of SARS-like coronaviruses.

    SciTech Connect

    Kapil, Sanjay; Oberst, R. D.; Bieker, Jill Marie; Tucker, Mark David; Souza, Caroline Ann; Williams, Cecelia Victoria

    2004-03-01

    Chemical disinfection and inactivation of viruses is largely understudied, but is very important especially in the case of highly infectious viruses. The purpose of this LDRD was to determine the efficacy of the Sandia National Laboratories developed decontamination formulations against Bovine Coronavirus (BCV) as a surrogate for the coronavirus that causes Severe Acute Respiratory Syndrome (SARS) in humans. The outbreak of SARS in late 2002 resulted from a highly infectious virus that was able to survive and remain infectious for extended periods. For this study, preliminary testing with Escherichia coli MS-2 (MS-2) and Escherichia coli T4 (T4) bacteriophages was conducted to develop virucidal methodology for verifying the inactivation after treatment with the test formulations following AOAC germicidal methodologies. After the determination of various experimental parameters (i.e. exposure, concentration) of the formulations, final testing was conducted on BCV. All experiments were conducted with various organic challenges (horse serum, bovine feces, compost) for results that more accurately represent field use condition. The MS-2 and T4 were slightly more resistant than BCV and required a 2 minute exposure while BCV was completely inactivated after a 1 minute exposure. These results were also consistent for the testing conducted in the presence of the various organic challenges indicating that the test formulations are highly effective for real world application.

  18. Developmental regulation of X-chromosome inactivation.

    PubMed

    Payer, Bernhard

    2016-08-01

    With the emergence of sex-determination by sex chromosomes, which differ in composition and number between males and females, appeared the need to equalize X-chromosomal gene dosage between the sexes. Mammals have devised the strategy of X-chromosome inactivation (XCI), in which one of the two X-chromosomes is rendered transcriptionally silent in females. In the mouse, the best-studied model organism with respect to XCI, this inactivation process occurs in different forms, imprinted and random, interspersed by periods of X-chromosome reactivation (XCR), which is needed to switch between the different modes of XCI. In this review, I describe the recent advances with respect to the developmental control of XCI and XCR and in particular their link to differentiation and pluripotency. Furthermore, I review the mechanisms, which influence the timing and choice, with which one of the two X-chromosomes is chosen for inactivation during random XCI. This has an impact on how females are mosaics with regard to which X-chromosome is active in different cells, which has implications on the severity of diseases caused by X-linked mutations.

  19. Kinetics of UV inactivation of wastewater bioflocs.

    PubMed

    Azimi, Y; Allen, D G; Farnood, R R

    2012-08-01

    Ultraviolet disinfection is a physical method of disinfecting secondary treated wastewaters. Bioflocs formed during secondary treatment harbor and protect microbes from exposure to ultraviolet (UV) light, and significantly decrease the efficiency of disinfection at high UV doses causing the tailing phenomena. However, the exact mechanism of tailing and the role of biofloc properties and treatment conditions are not widely understood. It is hypothesized that sludge bioflocs are composed of an easily disinfectable loose outer shell, and a physically stronger compact core inside that accounts for the tailing phenomena. Hydrodynamic shear stress was applied to the bioflocs to peel off the looser outer shell to isolate the cores. Biofloc and core samples were fractionated into narrow size distributions by sieving and their UV disinfection kinetics were determined and compared. The results showed that for bioflocs, the tailing level elevates as the biofloc size increases, showing greater resistance to disinfection. However, for the cores larger than 45μm, it was found that the UV inactivation curves overlap, and show very close to identical inactivation kinetics. Comparing bioflocs and cores of similar size fraction, it was found that in all cases cores were harder to disinfect with UV light, and showed a higher tailing level. This study suggests that physical structure of bioflocs plays a significant role in the UV inactivation kinetics. PMID:22608608

  20. Kinetics of UV inactivation of wastewater bioflocs.

    PubMed

    Azimi, Y; Allen, D G; Farnood, R R

    2012-08-01

    Ultraviolet disinfection is a physical method of disinfecting secondary treated wastewaters. Bioflocs formed during secondary treatment harbor and protect microbes from exposure to ultraviolet (UV) light, and significantly decrease the efficiency of disinfection at high UV doses causing the tailing phenomena. However, the exact mechanism of tailing and the role of biofloc properties and treatment conditions are not widely understood. It is hypothesized that sludge bioflocs are composed of an easily disinfectable loose outer shell, and a physically stronger compact core inside that accounts for the tailing phenomena. Hydrodynamic shear stress was applied to the bioflocs to peel off the looser outer shell to isolate the cores. Biofloc and core samples were fractionated into narrow size distributions by sieving and their UV disinfection kinetics were determined and compared. The results showed that for bioflocs, the tailing level elevates as the biofloc size increases, showing greater resistance to disinfection. However, for the cores larger than 45μm, it was found that the UV inactivation curves overlap, and show very close to identical inactivation kinetics. Comparing bioflocs and cores of similar size fraction, it was found that in all cases cores were harder to disinfect with UV light, and showed a higher tailing level. This study suggests that physical structure of bioflocs plays a significant role in the UV inactivation kinetics.

  1. Rabies virus inactivation by binary ethylenimine: new method for inactivated vaccine production.

    PubMed Central

    Larghi, O P; Nebel, A E

    1980-01-01

    The inactivation dynamics of rabies virus (PV strain) by binary ethylenimine, and the immunogenic properites and the stability of the vaccines prepared using this agent, were studied. Binary ethylenimine at a final concentration of 0.01 M was prepared wtih 2-bromoethylamine hydrobromide in alkaline solutions, either separately from or in suspensions of rabies virus propagated in BHK cells. The infectivity of virus suspensions containing more than 108 plaque-forming units per 0.1 ml was inactivated in 2 h when the inactivating agent was prepared before its addition to the suspensions, and in3 h when prepared directly in the suspensions. Liquid vaccines prepared in this manner and stored at different temperatures maintained potency for 1 month at 37 degrees C and for 6 months at 4 degrees C and 22 to 25 degrees C. Lyophilized vaccine maintained its potency for 6 months at the three temperatures. The inactivated vaccine mixed with aluminum or oil adjuvant at high dilutions protected guinea pigs against challenge. This safer procedure for rabies virus inactivation offers promise for the production of effective vaccines for the immunization of dogs and cattle. PMID:7358836

  2. Sodium channel inactivation in the crayfish giant axon. Must channels open before inactivating

    SciTech Connect

    Bean, B.P.

    1981-09-01

    Experiments on sodium channel inactivation kinetics were performed on voltage-clamped crayfish giant axons. The primary goals was to investigate whether channels must open before activating. Voltage-clamp artifacts were minimized by the use of low-sodium solutions and full series resistance compensation, and the spatial uniformity of the currents was checked with a closely spaced pair of electrodes used to measure local current densities. For membrane potentials between -40 and +40 mV, sodium currents decay to zero with a single exponential time-course. The time constant for decay is a steep function of membrane potential. The time-course of inactivation measured with the double-pulse method is very similar to the decay of current at the same potential. Steady-state inactivation curves measured with different test pulses are identical. The time-course of doubling pulse inactivation shows a lag that roughly correlates with the opening of sodium channels, but it is not strictly necessary for channels to open before inactivating. Measurements of the potential dependence of the integral of sodium conductance are also inconsistent with the simplest cases of models in which channels must open before activating.

  3. Determining the oligomer number of native GPCR using florescence correlation spectroscopy and drug-induced inactivation-reactivation.

    PubMed

    Teitler, Milt; Herrick-Davis, Katharine

    2014-01-01

    GPCRs are a major family of homologous proteins and are key mediators of the effects of numerous endogenous neurotransmitters, hormones, cytokines, therapeutic drugs, and drugs-of-abuse. Despite the enormous amount of research on the pharmacological and biochemical properties of GPCRs, there is surprisingly little information on GPCR dimer structure and function in primary cell culture or in vivo. We have used two novel approaches to develop methods to detect and study GPCR dimer function: FCS/PCH and "inactivation-reactivation". This review will focus on the data we have developed and our interpretations of those data. Using FCS/PCH 5-HT2C receptors have been detected directly and appear to exist as dimers, consistent with the inactivation-reactivation data on 5-HT7 and 5-HT2A receptors. Studies of the 5-HT7 and 5-HT2A serotonin receptors have revealed that binding of a pseudo-irreversible antagonist ("inactivator") to one of the orthosteric sites of a homodimer abolishes all receptor activity, and subsequent binding of a competitive antagonist to the orthosteric site of the second protomer releases the inactivator, allowing the receptor to return to an active state. This approach demonstrates allosteric crosstalk between protomers of native GPCR homodimers, indicating that GPCRs do exist and function as homodimers in both recombinant cells and rat primary astrocytes. This technique can be applied universally using intact recombinant or primary cells in culture, membrane homogenate preparations and, potentially, in vivo. This approach can be applied to heterodimers as well as homodimers and may aid in the development of novel drugs with heterodimer selectivity. PMID:25213307

  4. Irreversibility line in superconductor as line of constant shielding current density

    SciTech Connect

    Goemoery, F.; Takacs, S.; Holubar, T.

    1997-06-01

    The irreversibility of magnetic properties of superconductors is due to the existence of macroscopic shielding currents persisting for some period of time. The same currents offer nearly lossless electricity transport. Thus, the extent of magnetic irreversibility is directly proportional to the current-carrying capacity of a superconductor. Because the current-carrying capacity is an intrinsic property of the material, various experimental techniques should give the same irreversibility line corresponding to the same macroscopic shielding current density. Following this approach, the authors compared the irreversibility lines obtained from AC susceptibility measurement with those determined from quasistatic magnetization loops recorded with the help of a SQUID susceptometer. An additional parameter which has to be comparable is the electrical field characterizing the rate of change of the magnetic field. Fulfilling these conditions of equivalency, the authors found that it is possible to explain the irreversibility lines obtained by various techniques and at different conditions by the same physical model. They demonstrate that for the data, taken within two orders of magnitude for the current density and more than seven orders of magnitude for the electrical field, a consistent picture expressing all the observed features by the same model can be found. Measurements are presented from YBCO samples.

  5. Twisted partially pure spinors

    NASA Astrophysics Data System (ADS)

    Herrera, Rafael; Tellez, Ivan

    2016-08-01

    Motivated by the relationship between orthogonal complex structures and pure spinors, we define twisted partially pure spinors in order to characterize spinorially subspaces of Euclidean space endowed with a complex structure.

  6. “Slow” Voltage-Dependent Inactivation of CaV2.2 Calcium Channels Is Modulated by the PKC Activator Phorbol 12-Myristate 13-Acetate (PMA)

    PubMed Central

    Zhu, Lei; McDavid, Sarah; Currie, Kevin P. M.

    2015-01-01

    CaV2.2 (N-type) voltage-gated calcium channels (Ca2+ channels) play key roles in neurons and neuroendocrine cells including the control of cellular excitability, neurotransmitter / hormone secretion, and gene expression. Calcium entry is precisely controlled by channel gating properties including multiple forms of inactivation. “Fast” voltage-dependent inactivation is relatively well-characterized and occurs over the tens-to- hundreds of milliseconds timeframe. Superimposed on this is the molecularly distinct, but poorly understood process of “slow” voltage-dependent inactivation, which develops / recovers over seconds-to-minutes. Protein kinases can modulate “slow” inactivation of sodium channels, but little is known about if/how second messengers control “slow” inactivation of Ca2+ channels. We investigated this using recombinant CaV2.2 channels expressed in HEK293 cells and native CaV2 channels endogenously expressed in adrenal chromaffin cells. The PKC activator phorbol 12-myristate 13-acetate (PMA) dramatically prolonged recovery from “slow” inactivation, but an inactive control (4α-PMA) had no effect. This effect of PMA was prevented by calphostin C, which targets the C1-domain on PKC, but only partially reduced by inhibitors that target the catalytic domain of PKC. The subtype of the channel β-subunit altered the kinetics of inactivation but not the magnitude of slowing produced by PMA. Intracellular GDP-β-S reduced the effect of PMA suggesting a role for G proteins in modulating “slow” inactivation. We postulate that the kinetics of recovery from “slow” inactivation could provide a molecular memory of recent cellular activity and help control CaV2 channel availability, electrical excitability, and neurotransmission in the seconds-to-minutes timeframe. PMID:26222492

  7. Inactivation of the sodium channel. I. Sodium current experiments

    PubMed Central

    Bezanilla, F; Armstrong, CM

    1977-01-01

    Inactivation of sodium conductance has been studied in squid axons with voltage clamp techniques and with the enzyme pronase which selectively destroys inactivation. Comparison of the sodium current before and after pronase treatment shows a lag of several hundred microseconds in the onset of inactivation after depolarization. This lag can of several hundred microseconds in the onset of inactivation after polarization. This lag can also be demonstrated with double-pulse experiments. When the membrane potential is hyperpolarized to -140 mV before depolarization, both activation and inactivation are delayed. These findings suggest that inactivation occurs only after activation are delayed. These findings suggest that inactivation occurs only after activation; i.e. that the channels must open before they can inactivate. The time constant of inactivation measured with two pulses (τ(c)) is the same as the one measured from the decay of the sodium current during a single pulse (τ(h)). For large depolarizations, steady-state inactivation becomes more incomplete as voltage increases; but it is relatively complete and appears independent of voltage when determined with a two- pulse method. This result confirms the existence of a second open state for Na channels, as proposed by Chandler and Meves (1970. J. Physiol. [Lond.]. 211:653-678). The time constant of recovery from inactivation is voltage dependent and decreases as the membrane potential is made more negative. A model for Na channels is presented which has voltage-dependent transitions between the closed and open states, and a voltage-independent transition between the open and the inactivated state. In this model the voltage dependence of inactivation is a consequence of coupling to the activation process. PMID:591911

  8. High-Pressure Inactivation of Rotaviruses: Role of Treatment Temperature and Strain Diversity in Virus Inactivation.

    PubMed

    Araud, Elbashir; DiCaprio, Erin; Yang, Zhihong; Li, Xinhui; Lou, Fangfei; Hughes, John H; Chen, Haiqiang; Li, Jianrong

    2015-10-01

    Rotavirus (RV) is the major etiological agent of acute gastroenteritis in infants worldwide. Although high-pressure processing (HPP) is a popular method to inactivate enteric pathogens in food, the sensitivity of different virus strains within same species and serotype to HPP is variable. This study aimed to compare the barosensitivities of seven RV strains derived from four serotypes (serotype G1, strains Wa, Ku, and K8; serotype G2, strain S2; serotype G3, strains SA-11 and YO; and serotype G4, strain ST3) following high-pressure treatment. RV strains showed various responses to HPP based on the initial temperature and had different inactivation profiles. Ku, K8, S2, SA-11, YO, and ST3 showed enhanced inactivation at 4°C compared to 20°C. In contrast, strain Wa was not significantly impacted by the initial treatment temperature. Within serotype G1, strain Wa was significantly (P < 0.05) more resistant to HPP than strains Ku and K8. Overall, the resistance of the human RV strains to HPP at 4°C can be ranked as Wa > Ku = K8 > S2 > YO > ST3, and in terms of serotype the ranking is G1 > G2 > G3 > G4. In addition, pressure treatment of 400 MPa for 2 min was sufficient to eliminate the Wa strain, the most pressure-resistant RV, from oyster tissues. HPP disrupted virion structure but did not degrade viral protein or RNA, providing insight into the mechanism of viral inactivation by HPP. In conclusion, HPP is capable of inactivating RV at commercially acceptable pressures, and the efficacy of inactivation is strain dependent.

  9. High-Pressure Inactivation of Rotaviruses: Role of Treatment Temperature and Strain Diversity in Virus Inactivation

    PubMed Central

    Araud, Elbashir; DiCaprio, Erin; Yang, Zhihong; Li, Xinhui; Lou, Fangfei; Hughes, John H.; Chen, Haiqiang

    2015-01-01

    Rotavirus (RV) is the major etiological agent of acute gastroenteritis in infants worldwide. Although high-pressure processing (HPP) is a popular method to inactivate enteric pathogens in food, the sensitivity of different virus strains within same species and serotype to HPP is variable. This study aimed to compare the barosensitivities of seven RV strains derived from four serotypes (serotype G1, strains Wa, Ku, and K8; serotype G2, strain S2; serotype G3, strains SA-11 and YO; and serotype G4, strain ST3) following high-pressure treatment. RV strains showed various responses to HPP based on the initial temperature and had different inactivation profiles. Ku, K8, S2, SA-11, YO, and ST3 showed enhanced inactivation at 4°C compared to 20°C. In contrast, strain Wa was not significantly impacted by the initial treatment temperature. Within serotype G1, strain Wa was significantly (P < 0.05) more resistant to HPP than strains Ku and K8. Overall, the resistance of the human RV strains to HPP at 4°C can be ranked as Wa > Ku = K8 > S2 > YO > ST3, and in terms of serotype the ranking is G1 > G2 > G3 > G4. In addition, pressure treatment of 400 MPa for 2 min was sufficient to eliminate the Wa strain, the most pressure-resistant RV, from oyster tissues. HPP disrupted virion structure but did not degrade viral protein or RNA, providing insight into the mechanism of viral inactivation by HPP. In conclusion, HPP is capable of inactivating RV at commercially acceptable pressures, and the efficacy of inactivation is strain dependent. PMID:26187961

  10. Irreversibility in Ba sub 0. 625 K sub 0. 375 BiO sub 3

    SciTech Connect

    Shi, D. ); Ling, X.S. ); Xu, M. ); Fang, M.M. ); Luo, S.; Budnick, J.I. ); Dabrowski, B.; Hinks, D.G.; Richards, D.R.; Zheng, Y. )

    1991-02-01

    An irreversibility line has been identified in bulk Ba{sub 0.625}K{sub 0.375}BiO{sub 3}, which follows the relation 1{minus}{ital T}/{ital T}{sub {ital c}}{similar to}{ital H}{sup 2/3}. According to the flux-creep model, the magnetic irreversibility is caused by thermally activated flux creep and is closely connected with the field-induced resistive broadening. However, previous resistivity measurements showed that the Ba{sub 0.625}K{sub 0.375}BiO{sub 3} system did not exhibit resistive transition broadening. Thus, an alternative explanation based on the Josephson-coupling model is suggested to interpret the irreversibility observed in Ba{sub 0.625}K{sub 0.375}BiO{sub 3}.

  11. Mechanism for membrane electroporation irreversibility under high-intensity, ultrashort electrical pulse conditions.

    PubMed

    Joshi, R P; Schoenbach, K H

    2002-11-01

    An improved electroporation model is used to address membrane irreversibility under ultrashort electric pulse conditions. It is shown that membranes can survive a strong electric pulse and recover provided the pore distribution has a relatively large spread. If, however, the population consists predominantly of larger radii pores, then irreversibility can result. Physically, such a distribution could arise if pores at adjacent sites coalesce. The requirement of close proximity among the pore sites is more easily satisfied in smaller organelles than in outer cell membranes. Model predictions are in keeping with recent observations of cell damage to intracellular organelles (e.g., mitochondria), without irreversible shock at the outer membranes, by a nanosecond, high-intensity electric pulse. This mechanism also explains the greater damage from multiple electric shocks.

  12. Mechanism for membrane electroporation irreversibility under high-intensity, ultrashort electrical pulse conditions

    NASA Astrophysics Data System (ADS)

    Joshi, R. P.; Schoenbach, K. H.

    2002-11-01

    An improved electroporation model is used to address membrane irreversibility under ultrashort electric pulse conditions. It is shown that membranes can survive a strong electric pulse and recover provided the pore distribution has a relatively large spread. If, however, the population consists predominantly of larger radii pores, then irreversibility can result. Physically, such a distribution could arise if pores at adjacent sites coalesce. The requirement of close proximity among the pore sites is more easily satisfied in smaller organelles than in outer cell membranes. Model predictions are in keeping with recent observations of cell damage to intracellular organelles (e.g., mitochondria), without irreversible shock at the outer membranes, by a nanosecond, high-intensity electric pulse. This mechanism also explains the greater damage from multiple electric shocks.

  13. Inactivation of GAPDH as one mechanism of acrolein toxicity.

    PubMed

    Nakamura, Mizuho; Tomitori, Hideyuki; Suzuki, Takehiro; Sakamoto, Akihiko; Terui, Yusuke; Saiki, Ryotaro; Dohmae, Naoshi; Igarashi, Kazuei; Kashiwagi, Keiko

    2013-01-25

    We have recently reported that acrolein is more toxic than reactive oxygen species. Thus, the mechanism of cell toxicity by acrolein was studied using mouse mammary carcinoma FM3A cells. Acrolein-conjugated proteins were separated by gel electrophoresis with subsequent determination of their amino acid sequence, and it was found that glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was one of the major acrolein-conjugated proteins in cells. Acrolein interacted with cysteine-150 at the active site of GAPDH, and also with cysteine-282. When cells were treated with 8 μM acrolein, the activity of acrolein-conjugated GAPDH was greatly reduced, and the ATP content in cells was thus significantly reduced. In addition, it was shown that acrolein-conjugated GAPDH translocated to the nucleus, and the level of acetylated GAPDH and the number of TUNEL positive cells was increased, indicating that cell death is enhanced by acrolein-conjugated GAPDH. Inhibition of cell growth by acrolein was partially reversed when the cDNA encoding GAPDH was transformed into cells. These results indicate that inactivation of GAPDH is one mechanism that underlies cell toxicity caused by acrolein.

  14. Turnover of bacterial glutamine synthetase: oxidative inactivation precedes proteolysis.

    PubMed

    Levine, R L; Oliver, C N; Fulks, R M; Stadtman, E R

    1981-04-01

    We partially purified a preparation from Escherichia coli that proteolytically degrades the enzyme glutamine synthetase [L-glutamate:ammonia ligase (ADP-forming), EC 6.3.1.2]. The degradation is at least a two-step process. First, the glutamine synthetase undergoes an oxidative modification. This modification leads to loss of catalytic activity and also renders the protein susceptible to proteolytic attack in the second step. The oxidative step displays characteristics of a mixed-function oxidation, requiring both molecular oxygen and a reduced nucleotide. This step can also be catalyzed by a purified, mammalian cytochrome P-450 system, as well as by a model system consisting of ascorbic acid and oxygen. Catalase blocks this oxidative modification step. Thus, the overall process of proteolytic degradation can be observed only if care is taken to remove catalase activity from the extracts. The inactivation reaction is dependent on the state of adenylylation of the glutamine synthetase, suggesting that this a physiologically important reaction. If so, then mixed-function oxidases are now implicated in the process of intracellular protein turnover.

  15. Turnover of bacterial glutamine synthetase: oxidative inactivation precedes proteolysis.

    PubMed Central

    Levine, R L; Oliver, C N; Fulks, R M; Stadtman, E R

    1981-01-01

    We partially purified a preparation from Escherichia coli that proteolytically degrades the enzyme glutamine synthetase [L-glutamate:ammonia ligase (ADP-forming), EC 6.3.1.2]. The degradation is at least a two-step process. First, the glutamine synthetase undergoes an oxidative modification. This modification leads to loss of catalytic activity and also renders the protein susceptible to proteolytic attack in the second step. The oxidative step displays characteristics of a mixed-function oxidation, requiring both molecular oxygen and a reduced nucleotide. This step can also be catalyzed by a purified, mammalian cytochrome P-450 system, as well as by a model system consisting of ascorbic acid and oxygen. Catalase blocks this oxidative modification step. Thus, the overall process of proteolytic degradation can be observed only if care is taken to remove catalase activity from the extracts. The inactivation reaction is dependent on the state of adenylylation of the glutamine synthetase, suggesting that this a physiologically important reaction. If so, then mixed-function oxidases are now implicated in the process of intracellular protein turnover. Images PMID:6113590

  16. Irreversible binding of zomepirac to plasma protein in vitro and in vivo.

    PubMed Central

    Smith, P C; McDonagh, A F; Benet, L Z

    1986-01-01

    Zomepirac is a nonsteroidal anti-inflammatory drug recently withdrawn from use because of an unexplained high incidence of immunological reactions. It is metabolized in humans to a reactive, unstable acyl glucuronide which accumulates in plasma. Because of the similarity of zomepirac glucuronide to bilirubin glucuronide in structure and stability and the documented irreversible binding of bilirubin to albumin through its acyl glucuronide, we studied the reaction of zomepirac acyl glucuronide with albumin in vitro from pH 5 to 9 and in vivo in six healthy human volunteers who had received a single 100-mg oral dose of zomepirac. Irreversible binding of zomepirac to protein was determined by exhaustive washing of protein, followed by hydrolysis of bound zomepirac-protein adduct with base, extraction of the liberated drug, and chromatographic measurement. Irreversible binding was observed both in vitro and in vivo. The extent of binding in vitro was time- and pH-dependent. In vitro drug binding was also observed for the isomers of zomepirac glucuronide which were formed by intramolecular acyl migration. Irreversible binding in vivo correlated with overall exposure to zomepirac glucuronide when exposure was expressed as the area under the plasma concentration vs. time curve. When probenecid (500 mg, twice daily), which decreases the plasma clearance of zomepirac glucuronide, was administered concurrently with zomepirac, irreversible binding of zomepirac was increased. The nature of the zomepirac protein binding is probably covalent. Formation of irreversibly protein-bound zomepirac occurs via the acyl glucuronide as previously shown for bilirubin glucuronide, and the reaction may be general for other drugs that are metabolized to acyl glucuronides. PMID:3949982

  17. Detailed Modeling and Irreversible Transfer Process Analysis of a Multi-Element Thermoelectric Generator System

    NASA Astrophysics Data System (ADS)

    Xiao, Heng; Gou, Xiaolong; Yang, Suwen

    2011-05-01

    Thermoelectric (TE) power generation technology, due to its several advantages, is becoming a noteworthy research direction. Many researchers conduct their performance analysis and optimization of TE devices and related applications based on the generalized thermoelectric energy balance equations. These generalized TE equations involve the internal irreversibility of Joule heating inside the thermoelectric device and heat leakage through the thermoelectric couple leg. However, it is assumed that the thermoelectric generator (TEG) is thermally isolated from the surroundings except for the heat flows at the cold and hot junctions. Since the thermoelectric generator is a multi-element device in practice, being composed of many fundamental TE couple legs, the effect of heat transfer between the TE couple leg and the ambient environment is not negligible. In this paper, based on basic theories of thermoelectric power generation and thermal science, detailed modeling of a thermoelectric generator taking account of the phenomenon of energy loss from the TE couple leg is reported. The revised generalized thermoelectric energy balance equations considering the effect of heat transfer between the TE couple leg and the ambient environment have been derived. Furthermore, characteristics of a multi-element thermoelectric generator with irreversibility have been investigated on the basis of the new derived TE equations. In the present investigation, second-law-based thermodynamic analysis (exergy analysis) has been applied to the irreversible heat transfer process in particular. It is found that the existence of the irreversible heat convection process causes a large loss of heat exergy in the TEG system, and using thermoelectric generators for low-grade waste heat recovery has promising potential. The results of irreversibility analysis, especially irreversible effects on generator system performance, based on the system model established in detail have guiding significance for

  18. Eigenvalue analysis of an irreversible random walk with skew detailed balance conditions.

    PubMed

    Sakai, Yuji; Hukushima, Koji

    2016-04-01

    An irreversible Markov-chain Monte Carlo (MCMC) algorithm with skew detailed balance conditions originally proposed by Turitsyn et al. is extended to general discrete systems on the basis of the Metropolis-Hastings scheme. To evaluate the efficiency of our proposed method, the relaxation dynamics of the slowest mode and the asymptotic variance are studied analytically in a random walk on one dimension. It is found that the performance in irreversible MCMC methods violating the detailed balance condition is improved by appropriately choosing parameters in the algorithm. PMID:27176439

  19. An Irreversible Constitutive Law for Modeling the Delamination Process using Interface Elements

    NASA Technical Reports Server (NTRS)

    Goyal, Vinay K.; Johnson, Eric R.; Davila, Carlos G.; Jaunky, Navin; Ambur, Damodar (Technical Monitor)

    2002-01-01

    An irreversible constitutive law is postulated for the formulation of interface elements to predict initiation and progression of delamination in composite structures. An exponential function is used for the constitutive law such that it satisfies a multi-axial stress criterion for the onset of delamination, and satisfies a mixed mode fracture criterion for the progression of delamination. A damage parameter is included to prevent the restoration of the previous cohesive state between the interfacial surfaces. To demonstrate the irreversibility capability of the constitutive law, steady-state crack growth is simulated for quasi-static loading-unloading cycle of various fracture test specimens.

  20. An Irreversible Constitutive Law for Modeling the Delamination Process Using Interface Elements

    NASA Technical Reports Server (NTRS)

    Goyal, Vinay K.; Johnson, Eric R.; Davila, Carlos G.; Jaunky, Navin; Bushnell, Dennis M. (Technical Monitor)

    2002-01-01

    An irreversible constitutive law is postulated for the formulation of interface elements to predict initiation and progression of delamination in composite structures. An exponential function is used for the constitutive law such that it satisfies a multi-axial stress criterion for the onset of delamination, and satisfies a mixed mode fracture criterion for the progression of delamination. A damage parameter is included to prevent the restoration of the previous cohesive state between the interfacial surfaces. To demonstrate the irreversibility capability of the constitutive law, steady-state crack growth is simulated for quasi-static loading-unloading cycle of various fracture test specimens.

  1. Irreversibility in the microwave absorption of ceramic Y-Ba-Cu-O

    SciTech Connect

    Pakulis, E.J.

    1989-05-01

    We find that, for ceramic samples of YBa/sub 2/Cu/sub 3/O/sub x/, the microwave absorption in a dc magnetic field H and at a temperature T is different for field cooling than for zero field cooling, provided T is below a field-dependent irreversibility temperature. Since the microwaves probe only a layer at the surface of the sample, the irreversibility line in H-T space determined in this way is a property of the surface layer with a thickness given by the microwave skin depth. When the applied H is removed, we observe a remanent microwave absorption which decays logarithmically with time.

  2. Irreversibility and the Arrow of Time in a Quenched Quantum System.

    PubMed

    Batalhão, T B; Souza, A M; Sarthour, R S; Oliveira, I S; Paternostro, M; Lutz, E; Serra, R M

    2015-11-01

    Irreversibility is one of the most intriguing concepts in physics. While microscopic physical laws are perfectly reversible, macroscopic average behavior has a preferred direction of time. According to the second law of thermodynamics, this arrow of time is associated with a positive mean entropy production. Using a nuclear magnetic resonance setup, we measure the nonequilibrium entropy produced in an isolated spin-1/2 system following fast quenches of an external magnetic field. We experimentally demonstrate that it is equal to the entropic distance, expressed by the Kullback-Leibler divergence, between a microscopic process and its time reversal. Our result addresses the concept of irreversibility from a microscopic quantum standpoint. PMID:26588367

  3. The two-piston problem revisited: Generalization from reversible to irreversible expansion

    NASA Astrophysics Data System (ADS)

    Anacleto, Joaquim; Anacleto, Joaquim Alberto C.; Ferreira, J. M.

    2011-10-01

    We discuss the adiabatic two-piston problem for an ideal gas confined between two pistons of equal mass and extend recent work based on the reversible approximation. More realistic equations that account for the roles of the gas temperature and particle mass are applied to extend the analysis of the expansion of the gas from reversible to irreversible behavior to the limit of free expansion. The evolution of quantities, such as temperature, piston speed, adiabatic reversibility coefficients, and entropy, is obtained, and differences between the irreversible and the reversible solutions are investigated.

  4. The protein irreversible denaturation studied by means of the bending vibrational mode

    NASA Astrophysics Data System (ADS)

    Mallamace, Domenico; Corsaro, Carmelo; Vasi, Cirino; Vasi, Sebastiano; Dugo, Giacomo; Mallamace, Francesco

    2014-10-01

    We study by means of the infrared bending vibrational mode the microscopic mechanisms that are at the base of protein irreversible denaturation. In particular, we follow the thermal evolution of the Amide I and II vibrational modes of lysozyme residuals from ambient temperature toward the temperature of irreversible unfolding. Our results indicate that the thermal changes of the coupling, by means of the hydrogen bond, between hydration water molecules and the different chemical groups of the protein are the main microscopic mechanisms underlying the unfolding process.

  5. THE INACTIVATION OF TRYPSIN BY X-RAYS

    PubMed Central

    Clark, Harry; Northrop, John H.

    1925-01-01

    1. The inactivating effect of soft x-rays on trypsin in solutions of various degrees of concentration has been studied. 2. It has been found to run parallel with spontaneous heat inactivation. It is almost, if not entirely, confined to the free or active trypsin. 3. Under radiation of constant intensity, the inactivation follows the simple exponential law which indicates a monomolecular reaction. 4. Estimates have been made of the amount of ionization required to inactivate trypsin to half value in these experiments and in those of Hussey and Thompson, who employed the beta rays from Radium B and C. The close agreement corroborates the idea that the effect is a function of ionization only. 5. The nature of the process of inactivation is discussed; inactivation seems to result from electrical neutralization of the trypsin ion. PMID:19872235

  6. Inactivation kinetics of formaldehyde on N-acetyl-β-D-glucosaminidase from Nile tilapia (Oreochromis niloticus).

    PubMed

    Zhang, Wei-Ni; Bai, Ding-Ping; Lin, Xin-Yu; Chen, Qing-Xi; Huang, Xiao-Hong; Huang, Yi-Fan

    2014-04-01

    Formaldehyde is a widely used sanitizer in aquaculture in China, while the appropriate concentration is not available to be used effectively and without damage to tilapia much less to its reproductive function. N-acetyl-β-D-glucosaminidase (EC 3.2.1.52, NAGase), hydrolyzing the oligomers of N-acetyl-β-D-glucosamine into monomer, is proved to be correlated with reproduction of male animals. In this paper, NAGase from spermary of tilapia was chosen as the material to study the effects of formaldehyde on its activity in order to further investigate the effects of formaldehyde use on tilapia reproduction. The results showed the relationship between the residual enzyme activity and the concentration of formaldehyde was concentration dependent, and the IC50 value was estimated to be 3.2 ± 0.1 %. Appropriate concentration of formaldehyde leaded to competitive reversible inhibition on tilapia NAGase. Moreover, formaldehyde could reduce the thermal and pH stability of the enzyme. The inactivation kinetics of formaldehyde on the enzyme was studied using the kinetic method of substrate reaction. The inactivation model was setup, and the rate constants were determined. The results showed that the inactivation of formaldehyde on tilapia NAGase was a slow, reversible reaction with partially residual activity. The results will give some basis to determine the concentration of formaldehyde used in tilapia culture.

  7. Identification of lysine residue involved in inactivation of brain glutamate dehydrogenase isoproteins by o-phthalaldehyde.

    PubMed

    Ahn, J Y; Choi, S; Cho, S W

    1999-12-01

    Incubation of two types of glutamate dehydrogenase (GDH) isoproteins from bovine brain with o-phthalaldehyde resulted in a time-dependent loss of enzyme activity. The inactivation was partially prevented by preincubation of the GDH isoproteins with 2-oxoglutarate or NADH. Spectrophotometric studies indicated that the inactivation of GDH isoproteins with o-phthalaldehyde resulted in isoindole derivatives characterized by typical fluorescence emission spectra with a stoichiometry of one isoindole derivative per molecule of enzyme subunit. There were no differences between the two GDH isoproteins in sensitivities to inactivation by o-phthalaldehyde indicating that the microenvironmental structures of the GDH isoproteins are very similar to each other. Tryptic peptides of the isoproteins, modified with and without protection, identified a selective modification of one lysine as in the region containing the sequence L-Q-H-G-S-I-L-G-F-P-X-A-K for both GDH isoproteins. The symbol X indicates a position for which no phenylthiohydantoin-amino acid could be assigned. The missing residue, however, can be designated as an o-phthalaldehyde-labeled lysine since the sequences including the lysine residue in question have a complete identity with those of the other mammalian GDHs. Also, trypsin was unable to cleave the labeled peptide at this site. Both amino acid sequencing and compositional analysis identified Lys-306 as the site of o-phthalaldehyde binding within the brain GDH isoproteins. PMID:10607407

  8. Recurrent inactivating RASA2 mutations in melanoma.

    PubMed

    Arafeh, Rand; Qutob, Nouar; Emmanuel, Rafi; Keren-Paz, Alona; Madore, Jason; Elkahloun, Abdel; Wilmott, James S; Gartner, Jared J; Di Pizio, Antonella; Winograd-Katz, Sabina; Sindiri, Sivasish; Rotkopf, Ron; Dutton-Regester, Ken; Johansson, Peter; Pritchard, Antonia L; Waddell, Nicola; Hill, Victoria K; Lin, Jimmy C; Hevroni, Yael; Rosenberg, Steven A; Khan, Javed; Ben-Dor, Shifra; Niv, Masha Y; Ulitsky, Igor; Mann, Graham J; Scolyer, Richard A; Hayward, Nicholas K; Samuels, Yardena

    2015-12-01

    Analysis of 501 melanoma exomes identified RASA2, encoding a RasGAP, as a tumor-suppressor gene mutated in 5% of melanomas. Recurrent loss-of-function mutations in RASA2 were found to increase RAS activation, melanoma cell growth and migration. RASA2 expression was lost in ≥30% of human melanomas and was associated with reduced patient survival. These findings identify RASA2 inactivation as a melanoma driver and highlight the importance of RasGAPs in cancer. PMID:26502337

  9. Variance Components: Partialled vs. Common.

    ERIC Educational Resources Information Center

    Curtis, Ervin W.

    1985-01-01

    A new approach to partialling components is used. Like conventional partialling, this approach orthogonalizes variables by partitioning the scores or observations. Unlike conventional partialling, it yields a common component and two unique components. (Author/GDC)

  10. Methanol partial oxidation reformer

    DOEpatents

    Ahmed, S.; Kumar, R.; Krumpelt, M.

    1999-08-17

    A partial oxidation reformer is described comprising a longitudinally extending chamber having a methanol, water and an air inlet and an outlet. An igniter mechanism is near the inlets for igniting a mixture of methanol and air, while a partial oxidation catalyst in the chamber is spaced from the inlets and converts methanol and oxygen to carbon dioxide and hydrogen. Controlling the oxygen to methanol mole ratio provides continuous slightly exothermic partial oxidation reactions of methanol and air producing hydrogen gas. The liquid is preferably injected in droplets having diameters less than 100 micrometers. The reformer is useful in a propulsion system for a vehicle which supplies a hydrogen-containing gas to the negative electrode of a fuel cell. 7 figs.

  11. Methanol partial oxidation reformer

    DOEpatents

    Ahmed, Shabbir; Kumar, Romesh; Krumpelt, Michael

    1999-01-01

    A partial oxidation reformer comprising a longitudinally extending chamber having a methanol, water and an air inlet and an outlet. An igniter mechanism is near the inlets for igniting a mixture of methanol and air, while a partial oxidation catalyst in the chamber is spaced from the inlets and converts methanol and oxygen to carbon dioxide and hydrogen. Controlling the oxygen to methanol mole ratio provides continuous slightly exothermic partial oxidation reactions of methanol and air producing hydrogen gas. The liquid is preferably injected in droplets having diameters less than 100 micrometers. The reformer is useful in a propulsion system for a vehicle which supplies a hydrogen-containing gas to the negative electrode of a fuel cell.

  12. Methanol partial oxidation reformer

    DOEpatents

    Ahmed, Shabbir; Kumar, Romesh; Krumpelt, Michael

    2001-01-01

    A partial oxidation reformer comprising a longitudinally extending chamber having a methanol, water and an air inlet and an outlet. An igniter mechanism is near the inlets for igniting a mixture of methanol and air, while a partial oxidation catalyst in the chamber is spaced from the inlets and converts methanol and oxygen to carbon dioxide and hydrogen. Controlling the oxygen to methanol mole ratio provides continuous slightly exothermic partial oxidation reactions of methanol and air producing hydrogen gas. The liquid is preferably injected in droplets having diameters less than 100 micrometers. The reformer is useful in a propulsion system for a vehicle which supplies a hydrogen-containing gas to the negative electrode of a fuel cell.

  13. Methanol partial oxidation reformer

    DOEpatents

    Ahmed, S.; Kumar, R.; Krumpelt, M.

    1999-08-24

    A partial oxidation reformer is described comprising a longitudinally extending chamber having a methanol, water and an air inlet and an outlet. An igniter mechanism is near the inlets for igniting a mixture of methanol and air, while a partial oxidation catalyst in the chamber is spaced from the inlets and converts methanol and oxygen to carbon dioxide and hydrogen. Controlling the oxygen to methanol mole ratio provides continuous slightly exothermic partial oxidation reactions of methanol and air producing hydrogen gas. The liquid is preferably injected in droplets having diameters less than 100 micrometers. The reformer is useful in a propulsion system for a vehicle which supplies a hydrogen-containing gas to the negative electrode of a fuel cell. 7 figs.

  14. Oxygen partial pressure sensor

    DOEpatents

    Dees, D.W.

    1994-09-06

    A method for detecting oxygen partial pressure and an oxygen partial pressure sensor are provided. The method for measuring oxygen partial pressure includes contacting oxygen to a solid oxide electrolyte and measuring the subsequent change in electrical conductivity of the solid oxide electrolyte. A solid oxide electrolyte is utilized that contacts both a porous electrode and a nonporous electrode. The electrical conductivity of the solid oxide electrolyte is affected when oxygen from an exhaust stream permeates through the porous electrode to establish an equilibrium of oxygen anions in the electrolyte, thereby displacing electrons throughout the electrolyte to form an electron gradient. By adapting the two electrodes to sense a voltage potential between them, the change in electrolyte conductivity due to oxygen presence can be measured. 1 fig.

  15. Oxygen partial pressure sensor

    DOEpatents

    Dees, Dennis W.

    1994-01-01

    A method for detecting oxygen partial pressure and an oxygen partial pressure sensor are provided. The method for measuring oxygen partial pressure includes contacting oxygen to a solid oxide electrolyte and measuring the subsequent change in electrical conductivity of the solid oxide electrolyte. A solid oxide electrolyte is utilized that contacts both a porous electrode and a nonporous electrode. The electrical conductivity of the solid oxide electrolyte is affected when oxygen from an exhaust stream permeates through the porous electrode to establish an equilibrium of oxygen anions in the electrolyte, thereby displacing electrons throughout the electrolyte to form an electron gradient. By adapting the two electrodes to sense a voltage potential between them, the change in electrolyte conductivity due to oxygen presence can be measured.

  16. Inactivation of virus in solution by cold atmospheric pressure plasma: identification of chemical inactivation pathways

    NASA Astrophysics Data System (ADS)

    Aboubakr, Hamada A.; Gangal, Urvashi; Youssef, Mohammed M.; Goyal, Sagar M.; Bruggeman, Peter J.

    2016-05-01

    Cold atmospheric pressure plasma (CAP) inactivates bacteria and virus through in situ production of reactive oxygen and nitrogen species (RONS). While the bactericidal and virucidal efficiency of plasmas is well established, there is limited knowledge about the chemistry leading to the pathogen inactivation. This article describes a chemical analysis of the CAP reactive chemistry involved in the inactivation of feline calicivirus. We used a remote radio frequency CAP produced in varying gas mixtures leading to different plasma-induced chemistries. A study of the effects of selected scavengers complemented with positive control measurements of relevant RONS reveal two distinctive pathways based on singlet oxygen and peroxynitrous acid. The first mechanism is favored in the presence of oxygen and the second in the presence of air when a significant pH reduction is induced in the solution by the plasma. Additionally, smaller effects of the H2O2, O3 and \\text{NO}2- produced were also found. Identification of singlet oxygen-mediated 2-imidazolone/2-oxo-His (His  +14 Da)—an oxidative modification of His 262 comprising the capsid protein of feline calicivirus links the plasma induced singlet oxygen chemistry to viral inactivation.

  17. When X-inactivation meets pluripotency: an intimate rendezvous.

    PubMed

    Navarro, Pablo; Avner, Philip

    2009-06-01

    The integration of X-inactivation with development is a crucial aspect of this classical paradigm of epigenetic regulation. During early female mouse development, X-inactivation reprogramming occurs in pluripotent cells of the inner cell mass of the blastocyst and in pluripotent primordial germ cells. Here we discuss the developmental strategies which ensure the coupling of the regulation of X-inactivation to the acquisition of pluripotency through the regulation of the master of X-inactivation, the non-coding Xist gene, by the key factors which support pluripotency Nanog, Oct4 and Sox2.

  18. Sequential adsorption of an irreversibly adsorbed nonionic surfactant and an anionic surfactant at an oil/aqueous interface.

    PubMed

    Kirby, Stephanie M; Anna, Shelley L; Walker, Lynn M

    2015-04-14

    Aerosol-OT (AOT) and Tween 80 are two of the main surfactants in commercial dispersants used in response to oil spills. Understanding how multicomponent surfactant systems interact at oil/aqueous interfaces is crucial for improving both dispersant design and application efficacy. This is true of many multicomponent formulations; a lack of understanding of competition for the oil/water interface hinders formulation optimization. In this study, we have characterized the sequential adsorption behavior of AOT on squalane/aqueous interfaces that have been precoated with Tween 80. A microtensiometer is used to measure the dynamic interfacial tension of the system. Tween 80 either partially or completely irreversibly adsorbs to squalane/aqueous interfaces when rinsed with deionized water. These Tween 80 coated interfaces are then exposed to AOT. AOT adsorption increases with AOT concentration for all Tween 80 coverages, and the resulting steady-state interfacial tension values are interpreted using a Langmuir isotherm model. In the presence of 0.5 M NaCl, AOT adsorption significantly increases due to counterion charge screening of the negatively charged head groups. The presence of Tween 80 on the interface inhibits AOT adsorption, reducing the maximum surface coverage as compared to a clean interface. Tween 80 persists on the interface even after exposure to high concentrations of AOT.

  19. Irreversible transformation of ferromagnetic ordered stripe domains in single-shot infrared-pump/resonant-x-ray-scattering-probe experiments

    NASA Astrophysics Data System (ADS)

    Bergeard, Nicolas; Schaffert, Stefan; López-Flores, Víctor; Jaouen, Nicolas; Geilhufe, Jan; Günther, Christian M.; Schneider, Michael; Graves, Catherine; Wang, Tianhan; Wu, Benny; Scherz, Andreas; Baumier, Cédric; Delaunay, Renaud; Fortuna, Franck; Tortarolo, Marina; Tudu, Bharati; Krupin, Oleg; Minitti, Michael P.; Robinson, Joe; Schlotter, William F.; Turner, Joshua J.; Lüning, Jan; Eisebitt, Stefan; Boeglin, Christine

    2015-02-01

    The evolution of a magnetic domain structure upon excitation by an intense, femtosecond infrared (IR) laser pulse has been investigated using single-shot based time-resolved resonant x-ray scattering at the x-ray free electron laser LCLS. A well-ordered stripe domain pattern as present in a thin CoPd alloy film has been used as a prototype magnetic domain structure for this study. The fluence of the IR laser pump pulse was sufficient to lead to an almost complete quenching of the magnetization within the ultrafast demagnetization process taking place within the first few hundreds of femtoseconds following the IR laser pump pulse excitation. On longer time scales this excitation gave rise to subsequent irreversible transformations of the magnetic domain structure. Under our specific experimental conditions, it took about 2 ns before the magnetization started to recover. After about 5 ns the previously ordered stripe domain structure had evolved into a disordered labyrinth domain structure. Surprisingly, we observe after about 7 ns the occurrence of a partially ordered stripe domain structure reoriented into a novel direction. It is this domain structure in which the sample's magnetization stabilizes as revealed by scattering patterns recorded long after the initial pump-probe cycle. Using micromagnetic simulations we can explain this observation based on changes of the magnetic anisotropy going along with heat dissipation in the film.

  20. Sequential adsorption of an irreversibly adsorbed nonionic surfactant and an anionic surfactant at an oil/aqueous interface.

    PubMed

    Kirby, Stephanie M; Anna, Shelley L; Walker, Lynn M

    2015-04-14

    Aerosol-OT (AOT) and Tween 80 are two of the main surfactants in commercial dispersants used in response to oil spills. Understanding how multicomponent surfactant systems interact at oil/aqueous interfaces is crucial for improving both dispersant design and application efficacy. This is true of many multicomponent formulations; a lack of understanding of competition for the oil/water interface hinders formulation optimization. In this study, we have characterized the sequential adsorption behavior of AOT on squalane/aqueous interfaces that have been precoated with Tween 80. A microtensiometer is used to measure the dynamic interfacial tension of the system. Tween 80 either partially or completely irreversibly adsorbs to squalane/aqueous interfaces when rinsed with deionized water. These Tween 80 coated interfaces are then exposed to AOT. AOT adsorption increases with AOT concentration for all Tween 80 coverages, and the resulting steady-state interfacial tension values are interpreted using a Langmuir isotherm model. In the presence of 0.5 M NaCl, AOT adsorption significantly increases due to counterion charge screening of the negatively charged head groups. The presence of Tween 80 on the interface inhibits AOT adsorption, reducing the maximum surface coverage as compared to a clean interface. Tween 80 persists on the interface even after exposure to high concentrations of AOT. PMID:25798716

  1. MRI/DTI of the Brain Stem Reveals Reversible and Irreversible Disruption of the Baroreflex Neural Circuits: Clinical Implications.

    PubMed

    Su, Chia-Hao; Tsai, Ching-Yi; Chang, Alice Y W; Chan, Julie Y H; Chan, Samuel H H

    2016-01-01

    Baroreflex is the physiological mechanism for the maintenance of blood pressure and heart rate. Impairment of baroreflex is not a disease per se. However, depending on severity, the eventuality of baroreflex dysfunction varies from inconvenience in daily existence to curtailment of mobility to death. Despite universal acceptance, neuronal traffic within the contemporary neural circuits during the execution of baroreflex has never been visualized. By enhancing signal detection and fine-tuning the scanning parameters, we have successfully implemented tractographic analysis of the medulla oblongata in mice that allowed for visualization of connectivity between key brain stem nuclei in the baroreflex circuits. When viewed in conjunction with radiotelemetric analysis of the baroreflex, we found that under pathophysiological conditions when the disrupted connectivity between key nuclei in the baroreflex circuits was reversible, the associated disease condition (e.g. neurogenic hypertension) was amenable to remedial measures. Nevertheless, fatality ensues under pathological conditions (e.g. hepatic encephalopathy) when the connectivity between key substrates in the baroreflex circuits was irreversibly severed. MRI/DTI also prompted partial re-wiring of the contemporary circuit for baroreflex-mediated sympathetic vasomotor tone, and unearthed an explanation for the time lapse between brain death and the inevitable asystole signifying cardiac death that follows. PMID:27162554

  2. Irreversible change of electric conduction in ionic-liquid-gated (La,Sr)MnO3 thin films

    NASA Astrophysics Data System (ADS)

    Lee, Tae Kwon; Jung, Jong Hoon

    2016-10-01

    We have investigated the ionic-liquid-gating effect on electric conduction in (La0.8Sr0.2)MnO3(LSMO) thin films. The gating effect is significant for the LSMO thin films grown at low oxygen partial pressures. We observed that the channel resistance of LSMO was altered only for a positive gating voltage, not for a negative one, mainly through the changes of mobility rather than the carrier density. The increased sheet resistance at positive voltage does not return to the original value even after the removal of gating voltage as well as the application of a negative voltage. Through the Mn 2 p X-ray absorption, the increased resistance of LSMO after a positive voltage is found to be associated with the increase of the Mn3+ ions over Mn4+ ones. We proposed that oxygen vacancy and electrochemical reactions should play a role for the irreversible electric conduction in ionic-liquid-gated (La,Sr)MnO3 thin films.

  3. MRI/DTI of the Brain Stem Reveals Reversible and Irreversible Disruption of the Baroreflex Neural Circuits: Clinical Implications

    PubMed Central

    Su, Chia-Hao; Tsai, Ching-Yi; Chang, Alice Y.W.; Chan, Julie Y.H.; Chan, Samuel H.H.

    2016-01-01

    Baroreflex is the physiological mechanism for the maintenance of blood pressure and heart rate. Impairment of baroreflex is not a disease per se. However, depending on severity, the eventuality of baroreflex dysfunction varies from inconvenience in daily existence to curtailment of mobility to death. Despite universal acceptance, neuronal traffic within the contemporary neural circuits during the execution of baroreflex has never been visualized. By enhancing signal detection and fine-tuning the scanning parameters, we have successfully implemented tractographic analysis of the medulla oblongata in mice that allowed for visualization of connectivity between key brain stem nuclei in the baroreflex circuits. When viewed in conjunction with radiotelemetric analysis of the baroreflex, we found that under pathophysiological conditions when the disrupted connectivity between key nuclei in the baroreflex circuits was reversible, the associated disease condition (e.g. neurogenic hypertension) was amenable to remedial measures. Nevertheless, fatality ensues under pathological conditions (e.g. hepatic encephalopathy) when the connectivity between key substrates in the baroreflex circuits was irreversibly severed. MRI/DTI also prompted partial re-wiring of the contemporary circuit for baroreflex-mediated sympathetic vasomotor tone, and unearthed an explanation for the time lapse between brain death and the inevitable asystole signifying cardiac death that follows. PMID:27162554

  4. Dynamics of partial control.

    PubMed

    Sabuco, Juan; Sanjuán, Miguel A F; Yorke, James A

    2012-12-01

    Safe sets are a basic ingredient in the strategy of partial control of chaotic systems. Recently we have found an algorithm, the sculpting algorithm, which allows us to construct them, when they exist. Here we define another type of set, an asymptotic safe set, to which trajectories are attracted asymptotically when the partial control strategy is applied. We apply all these ideas to a specific example of a Duffing oscillator showing the geometry of these sets in phase space. The software for creating all the figures appearing in this paper is available as supplementary material. PMID:23278093

  5. Interference of the low-pH inactivated herpes simplex virus type 1 (HSV-1) strain HSZP with the early shutoff function of superinfecting HSV-1 strain KOS.

    PubMed

    Matis, J; Kúdelová, M; Rajcáni, J

    1999-03-01

    In former studies, we described that the HSZP strain of herpes simplex virus type 1 (HSV-1) was defective with respect to the early shutoff of host protein synthesis but was effective at interfering with the early shutoff function of the HSV-1 strain KOS, even when heat-inactivated or neutralized by antibody. However, the HSZP strain failed to interfere when inactivated with zinc ions or purified from cells treated with 2-deoxy-D-glucose. In this study, we provide evidence that the ability of the purified low-pH inactivated (citrate buffer, pH 3.0) and gel-filtered (Sephadex G-25) HSZP virions to adsorb host cells was not significantly affected. However, their ability to induce interference with the early shutoff function of the superinfecting HSV-1 strain KOS was restricted. In comparison with native virus, up to eight times more low-pH inactivated HSZP virions were needed to interfere with the shutoff by strain KOS. The interference was not due to exclusion of strain KOS by HSZP at the level of adsorption and/or penetration. The restriction was partially overcome by treatment of the cells with polyethylene glycol after adsorption of the low-pH inactivated HSZP virions. This observation indicates that the direct fusion of the virion envelope of low-pH inactivated HSZP with the plasma cell membrane was predominantly hampered.

  6. Survey of Impression Materials and Techniques in Fixed Partial Dentures among the Practitioners in India.

    PubMed

    Moldi, Arvind; Gala, Vimal; Puranik, Shivakumar; Karan, Smita; Deshpande, Sumit; Neela, Neelima

    2013-01-01

    Objective. Anecdotal evidence suggests that impression materials and techniques used in general dental practice for fixed partial dentures vary from those taught in dental schools. The aim of this survey was to integrate impression techniques evolved all over the years for fixed partial dentures and to know the techniques and materials which are used in the present day by the practitioners. Materials and Methods. A total of 1000 questionnaires were sent to various practitioners in India, out of which 807 questionnaires were filled. Results. The results showed that 84.8% of prosthodontists (65.56%, urban areas) use elastomeric impression materials as well as irreversible hydrocolloids and 15.2% use irreversible hydrocolloid only. Amongst other practitioners, 55.46% use irreversible hydrocolloid (45%, rural and semiurban areas) and 44.54% use elastomeric impression materials. Elastomeric impression technique practiced most commonly is putty reline with/without spacer (77.2%); other techniques are multiple-mix and monophase techniques. Conclusion. The ideal materials, technique, and armamentarium are required for the long-term success of the treatment for fixed partial denture. Also, if the ideal procedure is not followed, it will lead to a compromised fit of the final prosthesis and failure of the treatment.

  7. Inactivation of Toluene 2-Monooxygenase in Burkholderia cepacia G4 by Alkynes

    PubMed Central

    Yeager, Chris M.; Bottomley, Peter J.; Arp, Daniel J.; Hyman, Michael R.

    1999-01-01

    High concentrations of acetylene (10 to 50% [vol/vol] gas phase) were required to inhibit the growth of Burkholderia cepacia G4 on toluene, while 1% (vol/vol) (gas phase) propyne or 1-butyne completely inhibited growth. Low concentrations of longer-chain alkynes (C5 to C10) were also effective inhibitors of toluene-dependent growth, and 2- and 3-alkynes were more potent inhibitors than their 1-alkyne counterparts. Exposure of toluene-grown B. cepacia G4 to alkynes resulted in the irreversible loss of toluene- and o-cresol-dependent O2 uptake activities, while acetate- and 3-methylcatechol-dependent O2 uptake activities were unaffected. Toluene-dependent O2 uptake decreased upon the addition of 1-butyne in a concentration- and time-dependent manner. The loss of activity followed first-order kinetics, with apparent rate constants ranging from 0.25 min−1 to 2.45 min−1. Increasing concentrations of toluene afforded protection from the inhibitory effects of 1-butyne. Furthermore, oxygen, supplied as H2O2, was required for inhibition by 1-butyne. These results suggest that alkynes are specific, mechanism-based inactivators of toluene 2-monooxygenase in B. cepacia G4, although the simplest alkyne, acetylene, was relatively ineffective compared to longer alkynes. Alkene analogs of acetylene and propyne—ethylene and propylene—were not inactivators of toluene 2-monooxygenase activity in B. cepacia G4 but were oxidized to their respective epoxides, with apparent Ks and Vmax values of 39.7 μM and 112.3 nmol min−1 mg of protein−1 for ethylene and 32.3 μM and 89.2 nmol min−1 mg of protein−1 for propylene. PMID:9925593

  8. Mechanism of cysteine-dependent inactivation of aspartate/glutamate/cysteine sulfinic acid α-decarboxylases.

    PubMed

    Liu, Pingyang; Torrens-Spence, Michael P; Ding, Haizhen; Christensen, Bruce M; Li, Jianyong

    2013-02-01

    Animal aspartate decarboxylase (ADC), glutamate decarboxylase (GDC) and cysteine sulfinic acid decarboxylase (CSADC) catalyze the decarboxylation of aspartate, glutamate and cysteine sulfinic acid to β-alanine, γ-aminobutyric acid and hypotaurine, respectively. Each enzymatic product has been implicated in different physiological functions. These decarboxylases use pyridoxal 5-phosphate (PLP) as cofactor and share high sequence homology. Analysis of the activity of ADC in the presence of different amino determined that beta-alanine production from aspartate was diminished in the presence of cysteine. Comparative analysis established that cysteine also inhibited GDC and CSADC in a concentration-dependent manner. Spectral comparisons of free PLP and cysteine, together with ADC and cysteine, result in comparable spectral shifts. Such spectral shifts indicate that cysteine is able to enter the active site of the enzyme, interact with the PLP-lysine internal aldimine, form a cysteine-PLP aldimine and undergo intramolecular nucleophilic cyclization through its sulfhydryl group, leading to irreversible ADC inactivation. Cysteine is the building block for protein synthesis and a precursor of cysteine sulfinic acid that is the substrate of CSADC and therefore is present in many cells, but the presence of cysteine (at comparable concentrations to their natural substrates) apparently could severely inhibit ADC, CSADC and GDC activity. This raises an essential question as to how animal species prevent these enzymes from cysteine-mediated inactivation. Disorders of cysteine metabolism have been implicated in several neurodegenerative diseases. The results of our study should promote research in terms of mechanism by which animals maintain their cysteine homeostasis and possible relationship of cysteine-mediated GDC and CSADC inhibition in neurodegenerative disease development. PMID:22718265

  9. Mechanism of cysteine-dependent inactivation of aspartate/glutamate/cysteine sulfinic acid α-decarboxylases.

    PubMed

    Liu, Pingyang; Torrens-Spence, Michael P; Ding, Haizhen; Christensen, Bruce M; Li, Jianyong

    2013-02-01

    Animal aspartate decarboxylase (ADC), glutamate decarboxylase (GDC) and cysteine sulfinic acid decarboxylase (CSADC) catalyze the decarboxylation of aspartate, glutamate and cysteine sulfinic acid to β-alanine, γ-aminobutyric acid and hypotaurine, respectively. Each enzymatic product has been implicated in different physiological functions. These decarboxylases use pyridoxal 5-phosphate (PLP) as cofactor and share high sequence homology. Analysis of the activity of ADC in the presence of different amino determined that beta-alanine production from aspartate was diminished in the presence of cysteine. Comparative analysis established that cysteine also inhibited GDC and CSADC in a concentration-dependent manner. Spectral comparisons of free PLP and cysteine, together with ADC and cysteine, result in comparable spectral shifts. Such spectral shifts indicate that cysteine is able to enter the active site of the enzyme, interact with the PLP-lysine internal aldimine, form a cysteine-PLP aldimine and undergo intramolecular nucleophilic cyclization through its sulfhydryl group, leading to irreversible ADC inactivation. Cysteine is the building block for protein synthesis and a precursor of cysteine sulfinic acid that is the substrate of CSADC and therefore is present in many cells, but the presence of cysteine (at comparable concentrations to their natural substrates) apparently could severely inhibit ADC, CSADC and GDC activity. This raises an essential question as to how animal species prevent these enzymes from cysteine-mediated inactivation. Disorders of cysteine metabolism have been implicated in several neurodegenerative diseases. The results of our study should promote research in terms of mechanism by which animals maintain their cysteine homeostasis and possible relationship of cysteine-mediated GDC and CSADC inhibition in neurodegenerative disease development.

  10. Atmospheric pressure resistive barrier air plasma jet induced bacterial inactivation in aqueous environment

    NASA Astrophysics Data System (ADS)

    Thiyagarajan, Magesh; Sarani, Abdollah; Gonzales, Xavier

    2013-03-01

    An atmospheric pressure resistive barrier air plasma jet is designed to inactivate bacteria in aqueous media in direct and indirect exposure modes of treatment. The resistive barrier plasma jet is designed to operate at both dc and standard 50-60 Hz low frequency ac power input and the ambient air at 50% humidity level was used as the operating gas. The voltage-current characteristics of the plasma jet were analyzed and the operating frequency of the discharge was measured to be 20 kHz and the plasma power was measured to be 26 W. The plasma jet rotational temperatures (Trot) are obtained from the optical emission spectra, from the N2C-B(2+) transitions by matching the experimental spectrum results with the Spectra Air (SPECAIR) simulation spectra. The reactive oxygen and nitrogen species were measured using optical emission spectroscopy and gas analyzers, for direct and indirect treatment modes. The nitric oxides (NO) were observed to be the predominant long lived reactive nitrogen species produced by the plasma. Three different bacteria including Staphylococcus aureus (Gram-positive), Escherichia coli (Gram-negative), and Neisseria meningitidis (Gram-negative) were suspended in an aqueous media and treated by the resistive barrier air plasma jet in direct and indirect exposure modes. The results show that a near complete bacterial inactivation was achieved within 120 s for both direct and indirect plasma treatment of S. aureus and E. coli bacteria. Conversely, a partial inactivation of N. meningitidis was observed by 120 s direct plasma exposure and insignificant inactivation was observed for the indirect plasma exposure treatment. Plasma induced shifts in N. meningitidis gene expression was analyzed using pilC gene expression as a representative gene and the results showed a reduction in the expression of the pilC gene compared to untreated samples suggesting that the observed protection against NO may be regulated by other genes.

  11. Pulvinar inactivation disrupts selection of movement plans.

    PubMed

    Wilke, Melanie; Turchi, Janita; Smith, Katy; Mishkin, Mortimer; Leopold, David A

    2010-06-23

    The coordinated movement of the eyes and hands under visual guidance is an essential part of goal-directed behavior. Several cortical areas known to be involved in this process exchange projections with the dorsal aspect of the thalamic pulvinar nucleus, suggesting that this structure may play a central role in visuomotor behavior. Here, we used reversible inactivation to investigate the role of the dorsal pulvinar in the selection and execution of visually guided manual and saccadic eye movements in macaque monkeys. We found that unilateral pulvinar inactivation resulted in a spatial neglect syndrome accompanied by visuomotor deficits including optic ataxia during visually guided limb movements. Monkeys were severely disrupted in their visually guided behavior regarding space contralateral to the side of the injection in several domains, including the following: (1) target selection in both manual and oculomotor tasks, (2) limb usage in a manual retrieval task, and (3) spontaneous visual exploration. In addition, saccades into the ipsilesional field had abnormally short latencies and tended to overshoot their mark. None of the deficits could be explained by a visual field defect or primary motor deficit. These findings highlight the importance of the dorsal aspect of the pulvinar nucleus as a critical hub for spatial attention and selection of visually guided actions. PMID:20573910

  12. Protection against Japanese encephalitis by inactivated vaccines.

    PubMed

    Hoke, C H; Nisalak, A; Sangawhipa, N; Jatanasen, S; Laorakapongse, T; Innis, B L; Kotchasenee, S; Gingrich, J B; Latendresse, J; Fukai, K

    1988-09-01

    Encephalitis caused by Japanese encephalitis virus occurs in annual epidemics throughout Asia, making it the principal cause of epidemic viral encephalitis in the world. No currently available vaccine has demonstrated efficacy in preventing this disease in a controlled trial. We performed a placebo-controlled, blinded, randomized trial in a northern Thai province, with two doses of monovalent (Nakayama strain) or bivalent (Nakayama plus Beijing strains) inactivated, purified Japanese encephalitis vaccine made from whole virus derived from mouse brain. We examined the effect of these vaccines on the incidence and severity of Japanese encephalitis and dengue hemorrhagic fever, a disease caused by a closely related flavivirus. Between November 1984 and March 1985, 65,224 children received two doses of monovalent Japanese encephalitis vaccine (n = 21,628), bivalent Japanese encephalitis vaccine (n = 22,080), or tetanus toxoid placebo (n = 21,516), with only minor side effects. The cumulative attack rate for encephalitis due to Japanese encephalitis virus was 51 per 100,000 in the placebo group and 5 per 100,000 in each vaccine group. The efficacy in both vaccine groups combined was 91 percent (95 percent confidence interval, 70 to 97 percent). Attack rates for dengue hemorrhagic fever declined, but not significantly. The severity of cases of dengue was also reduced. We conclude that two doses of inactivated Japanese encephalitis vaccine, either monovalent or bivalent, protect against encephalitis due to Japanese encephalitis virus and may have a limited beneficial effect on the severity of dengue hemorrhagic fever.

  13. X-changing information on X inactivation

    SciTech Connect

    Barakat, Tahsin Stefan; Jonkers, Iris; Monkhorst, Kim; Gribnau, Joost

    2010-03-10

    In female somatic cells of mammalian species one X chromosome is inactivated to ensure dosage equality of X-encoded genes between females and males, during development and adulthood. X chromosome inactivation (XCI) involves various epigenetic mechanisms, including RNA mediated gene silencing in cis, DNA methylation, and changes in chromatin modifications and composition. XCI therefore provides an attractive paradigm to study epigenetic gene regulation in a more general context. The XCI process starts with counting of the number of X chromosomes present in a nucleus, and initiation of XCI follows if this number exceeds one per diploid genome. Recently, X-encoded RNF12 has been identified as a dose-dependent activator of XCI. In addition, other factors, including the pluripotency factors OCT4, SOX2 and Nanog, have been implicated to play a role in suppression of initiation of XCI. In this review, we highlight and explain these new and old findings in the context of a stochastic model for X chromosome counting and XCI initiation.

  14. X Chromosome Inactivation in Opioid Addicted Women

    PubMed Central

    Vousooghi, Nasim; Shirazi, Mitra-Sadat Sadat; Goodarzi, Ali; Abharian, Peyman Hassani; Zarrindast, Mohammad-Reza

    2015-01-01

    Introduction: X chromosome inactivation (XCI) is a process during which one of the two X chromosomes in female human is silenced leading to equal gene expression with males who have only one X chromosome. Here we have investigated XCI ratio in females with opioid addiction to see whether XCI skewness in women could be a risk factor for opioid addiction. Methods: 30 adult females meeting DSM IV criteria for opioid addiction and 30 control females with no known history of addiction were included in the study. Digested and undigested DNA samples which were extracted from blood were analyzed after amplification of the polymorphic androgen receptor (AR) gene located on the X chromosome. XCI skewness was studied in 3 ranges: 50:50–64:36 (random inactivation), 65:35–80:20 (moderately skewed) and >80:20 (highly skewed). Results: XCI from informative females in control group was 63% (N=19) random, 27% (N=8) moderately skewed and 10% (N=3) highly skewed. Addicted women showed 57%, 23% and 20%, respectively. The distribution and frequency of XCI status in women with opioid addiction was not significantly different from control group (P=0.55). Discussion: Our data did not approve our hypothesis of increased XCI skewness among women with opioid addiction or unbalanced (non-random) expression of genes associated with X chromosome in female opioid addicted subjects. PMID:26904175

  15. Inactivation of a subtilisin in colloidal systems.

    PubMed

    Maste, M C; Rinia, H A; Brands, C M; Egmond, M R; Norde, W

    1995-10-25

    The aim of the present study is to establish the relation between the inactivation of the proteolytic enzyme Savinase and its adsorption at different types of solid-liquid interfaces. The loss of activity has been determined both in solution and in the presence of colloidal particles, which provide a surface area for adsorption of 25% of the enzyme population. Analysis of the remaining solution at different periods of incubation of the various systems shows that the intact protein is converted into autolytic degradation products at the expense of biological activity. The different particles, however, deactivate the enzymes to a different extent. Under the experimental conditions the half-life of the enzymatic activity in solution is 3.5 hours. In the presence of particles that have hydrophobic surface properties (teflon- or polystyrene latex) the half-life is reduced to 0.7 hours. On the contrary, hydrophilic silica particles stabilize the enzyme against autolysis as compared to the inactivation in solution. Polystyrene latex particles which are chemically grafted with short poly(ethylene oxide) chains ([EO]8) are, for steric reasons, also mild with respect to the reduction of enzymatic stability. It is thus concluded that the type of surface determines the mode in which the enzyme is adsorbed on a particle which, in turn, affects the autocatalytic rate.

  16. IL26 gene inactivation in Equidae.

    PubMed

    Shakhsi-Niaei, M; Drögemüller, M; Jagannathan, V; Gerber, V; Leeb, T

    2013-12-01

    Interleukin-26 (IL26) is a member of the IL10 cytokine family. The IL26 gene is located between two other well-known cytokines genes of this family encoding interferon-gamma (IFNG) and IL22 in an evolutionary conserved gene cluster. In contrast to humans and most other mammals, mice lack a functional Il26 gene. We analyzed the genome sequences of other vertebrates for the presence or absence of functional IL26 orthologs and found that the IL26 gene has also become inactivated in several equid species. We detected a one-base pair frameshift deletion in exon 2 of the IL26 gene in the domestic horse (Equus caballus), Przewalski horse (Equus przewalskii) and donkey (Equus asinus). The remnant IL26 gene in the horse is still transcribed and gives rise to at least five alternative transcripts. None of these transcripts share a conserved open reading frame with the human IL26 gene. A comparative analysis across diverse vertebrates revealed that the IL26 gene has also independently been inactivated in a few other mammals, including the African elephant and the European hedgehog. The IL26 gene thus appears to be highly variable, and the conserved open reading frame has been lost several times during mammalian evolution.

  17. Photodynamic inactivation of pathogens causing infectious keratitis

    NASA Astrophysics Data System (ADS)

    Simon, Carole; Wolf, G.; Walther, M.; Winkler, K.; Finke, M.; Hüttenberger, D.; Bischoff, Markus; Seitz, B.; Cullum, J.; Foth, H.-J.

    2014-03-01

    The increasing prevalence of antibiotic resistance requires new approaches also for the treatment of infectious keratitis. Photodynamic Inactivation (PDI) using the photosensitizer (PS) Chlorin e6 (Ce6) was investigated as an alternative to antibiotic treatment. An in-vitro cornea model was established using porcine eyes. The uptake of Ce6 by bacteria and the diffusion of the PS in the individual layers of corneal tissue were investigated by fluorescence. After removal of the cornea's epithelium Ce6-concentrations < 1 mM were sufficient to reach a penetration depth of 500 μm. Liquid cultures of microorganisms were irradiated using a specially constructed illumination chamber made of Spectralon(R) (reflectance: 99 %), which was equipped with high power light emitting diodes (λ = 670 nm). Clinical isolates of Staphylococcus aureus (SA) and Pseudomonas aeruginosa (PA) from keratitis patients were tested in liquid culture against different concentrations of Ce6 (1 - 512 μM) using 10 minutes irradiation (E = 18 J/cm2 ). This demonstrated that a complete inactivation of the pathogen strains were feasible whereby SA was slightly more susceptible than PA. 3909 mutants of the Keio collection of Escherichia coli (E.coli) were screened for potential resistance factors. The sensitive mutants can be grouped into three categories: transport mutants, mutants in lipopolysaccharide synthesis and mutants in the bacterial SOS-response. In conclusion PDI is seen as a promising therapy concept for infectious keratitis.

  18. A tyrosine-reactive irreversible inhibitor for glutathione S-transferase Pi (GSTP1).

    PubMed

    Crawford, L A; Weerapana, E

    2016-05-24

    Glutathione S-transferase Pi (GSTP1) mediates cellular defense against reactive electrophiles. Here, we report LAS17, a dichlorotriazine-containing compound that irreversibly inhibits GSTP1 and is selective for GSTP1 within cellular proteomes. Mass spectrometry and mutational studies identified Y108 as the site of modification, providing a unique mode of GSTP1 inhibition. PMID:27113843

  19. Thermodynamics of Irreversible Processes. Physical Processes in Terrestrial and Aquatic Ecosystems, Transport Processes.

    ERIC Educational Resources Information Center

    Levin, Michael; Gallucci, V. F.

    These materials were designed to be used by life science students for instruction in the application of physical theory to ecosystem operation. Most modules contain computer programs which are built around a particular application of a physical process. This module describes the application of irreversible thermodynamics to biology. It begins with…

  20. A comparison of the anesthetic efficacy of articaine and lidocaine in patients with irreversible pulpitis.

    PubMed

    Tortamano, Isabel Peixoto; Siviero, Marcelo; Costa, Carina Gisele; Buscariolo, Inês Aparecida; Armonia, Paschoal Laércio

    2009-02-01

    The purpose of the present study was to compare the anesthetic efficacy of 4% articaine with 1:100,000 epinephrine with that of 2% lidocaine with 1:100,000 epinephrine during pulpectomy in patients with irreversible pulpitis in mandibular posterior teeth. Forty volunteers, patients with irreversible pulpitis admitted to the Emergency Center of the School of Dentistry at the University of São Paulo, randomly received a conventional inferior alveolar nerve block containing 3.6 mL of either 4% articaine with 1:100,000 epinephrine or 2% lidocaine with 1:100,000 epinephrine. During the subsequent pulpectomy, we recorded the patients' subjective assessments of lip anesthesia, the absence/presence of pulpal anesthesia through electric pulp stimulation, and the absence/presence of pain through a verbal analogue scale. All tested patients reported lip anesthesia after the application of either inferior alveolar nerve block. Regarding pulpal anesthesia success as measured with the pulp tester, the lidocaine solution had a higher success rate (70%) than the articaine solution (65%). For patients reporting none or mild pain during pulpectomy, the success rate of the articaine solution (65%) was higher than that of the lidocaine solution (45%). Yet, none of the observed differences between articaine and lidocaine were statistically significant. Apparently, therefore, both local anesthetic solutions had similar effects on the patients with irreversible pulpitis in mandibular posterior teeth. Neither of the solutions, however, resulted in an effective pain control during irreversible pulpitis treatments.

  1. Flux pinning and irreversibility in YBa sub 2 Cu sub 3 O sub 7 superconducting crystal

    SciTech Connect

    Sagdahl, L.T.; Gjolmesli, S.; Laegreid, T.; Fossheim, K.; Assmus, W. )

    1990-10-01

    Extensive ac-magnetic-permeability studies have been carried out on the irreversible behavior and flux dynamics in superconducting YBa{sub 2}Cu{sub 3}O{sub 7} crystals in external magnetic fields up to 7.5 T. From the analysis the irreversibility line {ital B}{sup *}({ital T}{sup *}) is found to scale as (1{minus}{ital T}{sup *}/{Tc}){sup {ital n}}, where {ital n}{approx equal}1.5 for fields higher than 1.5 T, and {ital n}{approx equal}1 for lower fields. The exponents are sensitive to the choice of {Tc}. The frequency dependence of the irreversibility temperature is carefully studied in the low-frequency region from 10 to 10{sup 5} Hz and is found to be logarithmic in the whole frequency range. The small logarithmic slope increases with increasing external field. The analysis is done within a flux-creep picture using a single-relaxation-time form for the complex permeability, and a vortex-glass model. The peak in the imaginary part of the permeability in different dc fields is successfully fitted to the predictions of the flux-creep model. A dynamical screening length is needed to account for the observed frequency dependence of the irreversibility temperature.

  2. 50 CFR 402.09 - Irreversible or irretrievable commitment of resources.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE); ENDANGERED SPECIES COMMITTEE REGULATIONS SUBCHAPTER A INTERAGENCY COOPERATION-ENDANGERED SPECIES ACT OF 1973, AS AMENDED General § 402.09 Irreversible or... proposed species or proposed critical habitat under section 7(a)(4) of the Act....

  3. 50 CFR 402.09 - Irreversible or irretrievable commitment of resources.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE); ENDANGERED SPECIES COMMITTEE REGULATIONS SUBCHAPTER A INTERAGENCY COOPERATION-ENDANGERED SPECIES ACT OF 1973, AS AMENDED General § 402.09 Irreversible or... proposed species or proposed critical habitat under section 7(a)(4) of the Act....

  4. 50 CFR 402.09 - Irreversible or irretrievable commitment of resources.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE); ENDANGERED SPECIES COMMITTEE REGULATIONS SUBCHAPTER A INTERAGENCY COOPERATION-ENDANGERED SPECIES ACT OF 1973, AS AMENDED General § 402.09 Irreversible or... proposed species or proposed critical habitat under section 7(a)(4) of the Act....

  5. 50 CFR 402.09 - Irreversible or irretrievable commitment of resources.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE); ENDANGERED SPECIES COMMITTEE REGULATIONS SUBCHAPTER A INTERAGENCY COOPERATION-ENDANGERED SPECIES ACT OF 1973, AS AMENDED General § 402.09 Irreversible or... proposed species or proposed critical habitat under section 7(a)(4) of the Act....

  6. 50 CFR 402.09 - Irreversible or irretrievable commitment of resources.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE); ENDANGERED SPECIES COMMITTEE REGULATIONS SUBCHAPTER A INTERAGENCY COOPERATION-ENDANGERED SPECIES ACT OF 1973, AS AMENDED General § 402.09 Irreversible or... proposed species or proposed critical habitat under section 7(a)(4) of the Act....

  7. Seleninate in Place of Phosphate: Irreversible Inhibition of Protein Tyrosine Phosphatases

    SciTech Connect

    Abdo, Mohannad; Liu, Sijiu; Zhou, Bo; Walls, Chad D.; Wu, Li; Knapp, Spencer; Zhang, Zhong-Yin

    2009-02-16

    A homotyrosine based seleninic acid irreversibly inhibits protein tyrosine phosphatases by forming a covalent selenosulfide linkage with the active site cysteine sulfhydryl specifically. The details of the event are revealed by model synthetic studies and by kinetic, mass spectrometric, and crystallographic characterization.

  8. A tyrosine-reactive irreversible inhibitor for glutathione S-transferase Pi (GSTP1).

    PubMed

    Crawford, L A; Weerapana, E

    2016-05-24

    Glutathione S-transferase Pi (GSTP1) mediates cellular defense against reactive electrophiles. Here, we report LAS17, a dichlorotriazine-containing compound that irreversibly inhibits GSTP1 and is selective for GSTP1 within cellular proteomes. Mass spectrometry and mutational studies identified Y108 as the site of modification, providing a unique mode of GSTP1 inhibition.

  9. General Law of Electromagnetic Radiation Conversion Efficiency in Systems with Linear and Non-Linear Irreversibility

    NASA Astrophysics Data System (ADS)

    Chukova, Yu. P.

    2011-12-01

    It is shown, that the efficiency of conversion of solar radiation obeys the same law in alive and nonliving (technical) systems. For different processes in alive systems the evolution has selected different ranges of solar intensity and different conditions of irreversibility.

  10. Irreversible Loss of Vision in a Child due to Occipital Infarction after Gastroenteritis.

    PubMed

    Mansour, Ahmad M; Hasbini, Dana; Younis, Muhammad H; Bhatti, M Tariq

    2015-01-01

    A 2½-year-old girl developed a bilateral occipital infarct following severe gastroenteritis with bilateral vision of light perception. Evaluations for sickle cell anemia, hemolytic anemia and coagulopathies were negative. Cortical blindness is an uncommon but dramatic complication of gastroenteritis, hence the need of prompt hydration and other supportive measures to avoid irreversible visual loss or mental sequela.

  11. Irreversible Loss of Vision in a Child due to Occipital Infarction after Gastroenteritis

    PubMed Central

    Mansour, Ahmad M.; Hasbini, Dana; Younis, Muhammad H.; Bhatti, M. Tariq

    2015-01-01

    A 2½-year-old girl developed a bilateral occipital infarct following severe gastroenteritis with bilateral vision of light perception. Evaluations for sickle cell anemia, hemolytic anemia and coagulopathies were negative. Cortical blindness is an uncommon but dramatic complication of gastroenteritis, hence the need of prompt hydration and other supportive measures to avoid irreversible visual loss or mental sequela. PMID:25960732

  12. Expansion Work without the External Pressure and Thermodynamics in Terms of Quasistatic Irreversible Processes

    ERIC Educational Resources Information Center

    Schmidt-Rohr, Klaus

    2014-01-01

    We demonstrate that the formula for irreversible expansion work in most chemical thermodynamics textbooks does not apply during the expansion process. Instead of the "external pressure" P[subscript ext], the pressure P[subscript sys,mb] on the piston or other moving boundary (hence the subscript mb), which is nearly equal to the system…

  13. Irreversible Capacities of Graphite in Low Temperature Electrolytes for Lithium-Ion Batteries

    NASA Technical Reports Server (NTRS)

    Ratnakumar, B.; Smart, M.; Surampudi, S.; Wang, Y.; Zhang, X.; Greenbaum, S.; Hightower, A.; Ahn, C.; Fultz, B.

    1999-01-01

    Carbonaceous anode materials in lithium ion rechargeable cells experience irreversible capacity, mainly due to a consumption of lithium in the formation of surface passive films. The stability and kinetics of lithium intercalation into the carbon anodes are dictated by these films.

  14. Partial hue-matching.

    PubMed

    Logvinenko, Alexander D; Beattie, Lesley L

    2011-01-01

    It is widely believed that color can be decomposed into a small number of component colors. Particularly, each hue can be described as a combination of a restricted set of component hues. Methods, such as color naming and hue scaling, aim at describing color in terms of the relative amount of the component hues. However, there is no consensus on the nomenclature of component hues. Moreover, the very notion of hue (not to mention component hue) is usually defined verbally rather than perceptually. In this paper, we make an attempt to operationalize such a fundamental attribute of color as hue without the use of verbal terms. Specifically, we put forth a new method--partial hue-matching--that is based on judgments of whether two colors have some hue in common. It allows a set of component hues to be established objectively, without resorting to verbal definitions. Specifically, the largest sets of color stimuli, all of which partially match each other (referred to as chromaticity classes), can be derived from the observer's partial hue-matches. A chromaticity class proves to consist of all color stimuli that contain a particular component hue. Thus, the chromaticity classes fully define the set of component hues. Using samples of Munsell papers, a few experiments on partial hue-matching were carried out with twelve inexperienced normal trichromatic observers. The results reinforce the classical notion of four component hues (yellow, blue, red, and green). Black and white (but not gray) were also found to be component colors. PMID:21742961

  15. Partial knee replacement

    MedlinePlus

    ... You will need to understand what surgery and recovery will be like. Partial knee arthroplasty may be a good choice if you have arthritis in only one side or part of the knee and: You are older, thin, and not very active. You do not ...

  16. Effect of hydraulically reversible and hydraulically irreversible fouling on the removal of MS2 and φX174 bacteriophage by an ultrafiltration membrane.

    PubMed

    ElHadidy, Ahmed M; Peldszus, Sigrid; Van Dyke, Michele I

    2014-09-15

    The effect of membrane fouling on the removal of enteric virus surrogates MS2 and φX174 bacteriophage by an ultrafiltration membrane was assessed under simulated full-scale drinking water treatment operating conditions. Filtration experiments of up to 8 days using either river or lake water ascertained how the membrane fouling layer affected virus removal. Organic carbon fractionation techniques identified potential foulants, including biopolymers, in the feed water and in the permeate. Hydraulically irreversible fouling could greatly improve the removal of both viruses at moderate and severe fouling conditions by up to 2.5 logs. Hydraulically reversible fouling increased virus removal only slightly, and increased removal of >0.5 log for both phage were only obtained under severe fouling conditions. The increase in virus removal due to irreversible and reversible fouling differed between the two water sources. As the degree of fouling increased, differences between the removal of the two phage decreased. Maintenance cleaning partially removed membrane foulants, however virus removal following maintenance cleaning was lower than that of the fouled membrane, it remained higher than that of the clean membrane.

  17. Pinning and irreversibility in superconducting bulk MgB2 with added nanodiamonds

    NASA Astrophysics Data System (ADS)

    Gupta, Anurag; Narlikar, A. V.

    2009-12-01

    Resistance, R(T), and magnetization, M(B), studies on superconducting bulk MgB2 samples containing nanodiamonds (ND) as additives (wt% of ND: x = 0%, 1%, 3%, 5%, 7% and 10%) were recently published in two articles (Vajpayee et al 2007 Supercond. Sci. Technol. 20 S155, Vajpayee et al 2008 J. Appl. Phys. 103 07C708). The main observations reported were significant improvements in the critical current density Jc(B), irreversibility line Birr(T) and upper critical field Bc2(T) with ND addition. However, a closer look shows that as regards the potential of this technologically important material at higher magnetic fields and temperatures, there is still a lot of room for improvement. With that in mind we revisit the R(T) and M(B) data and analyze them, in the present work. We show that, despite ND addition, Jc depends strongly on B in the high field region and tends to vanish at irreversibility lines that lie deep, i.e. at around 0.3 Bc2(T), in the B-T phase diagram. The irreversibility lines, determined by R(T) \\to 0 in the presence of B, are found to lie at around 0.5 Bc2(T) in the phase diagram. These results for pinning and irreversibility lines are discussed in the light of various models such as those of surface sheath superconductivity, magnetically introduced percolation in polycrystalline MgB2, thermally assisted flux motion (TAFM) and a modified flux line shear mechanism. Our analysis hints at TAFM and weak pinning channels with distributed superconducting properties percolating in our samples determining the irreversibility and pinning properties.

  18. Using irreversible compression in digital radiology: a preliminary study of the opinions of radiologists

    NASA Astrophysics Data System (ADS)

    Seeram, Euclid

    2006-03-01

    The large volumes of digital images produced by digital imaging modalities in Radiology have provided the motivation for the development of picture archiving and communication systems (PACS) in an effort to provide an organized mechanism for digital image management. The development of more sophisticated methods of digital image acquisition (Multislice CT and Digital Mammography, for example), as well as the implementation and performance of PACS and Teleradiology systems in a health care environment, have created challenges in the area of image compression with respect to storing and transmitting digital images. Image compression can be reversible (lossless) or irreversible (lossy). While in the former, there is no loss of information, the latter presents concerns since there is a loss of information. This loss of information from diagnostic medical images is of primary concern not only to radiologists, but also to patients and their physicians. In 1997, Goldberg pointed out that "there is growing evidence that lossy compression can be applied without significantly affecting the diagnostic content of images... there is growing consensus in the radiologic community that some forms of lossy compression are acceptable". The purpose of this study was to explore the opinions of expert radiologists, and related professional organizations on the use of irreversible compression in routine practice The opinions of notable radiologists in the US and Canada are varied indicating no consensus of opinion on the use of irreversible compression in primary diagnosis, however, they are generally positive on the notion of the image storage and transmission advantages. Almost all radiologists are concerned with the litigation potential of an incorrect diagnosis based on irreversible compressed images. The survey of several radiology professional and related organizations reveals that no professional practice standards exist for the use of irreversible compression. Currently, the

  19. [Determination of irreversibility of clinical brain death. Electroencephalography and evoked potentials].

    PubMed

    Buchner, H; Ferbert, A

    2016-02-01

    Principally, in the fourth update of the rules for the procedure to finally determine the irreversible cessation of function of the cerebrum, the cerebellum and the brainstem, the importance of an electroencephalogram (EEG), somatosensory evoked potentials (SEP) and brainstem auditory evoked potentials (BAEP) are confirmed. This paper presents the reliability and validity of the electrophysiological diagnosis, discusses the amendments in the fourth version of the guidelines and introduces the practical application, problems and sources of error.An EEG is the best established supplementary diagnostic method for determining the irreversibility of clinical brain death syndrome. It should be noted that residual brain activity can often persist for many hours after the onset of brain death syndrome, particularly in patients with primary brainstem lesions. The derivation and analysis of an EEG requires a high level of expertise to be able to safely distinguish artefacts from primary brain activity. The registration of EEGs to demonstrate the irreversibility of clinical brain death syndrome is extremely time consuming.The BAEPs can only be used to confirm the irreversibility of brain death syndrome in serial examinations or in the rare cases of a sustained wave I or sustained waves I and II. Very often, an investigation cannot be reliably performed because of existing sound conduction disturbances or failure of all potentials even before the onset of clinical brain death syndrome. This explains why BAEPs are only used in exceptional cases.The SEPs of the median nerve can be very reliably derived, are technically simple and with few sources of error. A serial investigation is not required and the time needed for examination is short. For these reasons SEPs are given preference over EEGs and BAEPs for establishing the irreversibility of clinical brain death syndrome. PMID:26785843

  20. Role of cysteines in the activation and inactivation of brewers' yeast pyruvate decarboxylase investigated with a PDC1-PDC6 fusion protein.

    PubMed

    Zeng, X; Farrenkopf, B; Hohmann, S; Dyda, F; Furey, W; Jordan, F

    1993-03-16

    Possible roles of the Cys side chains in the activation and inactivation mechanisms of brewers' yeast pyruvate decarboxylase were investigated by comparing the behavior of the tetrameric enzyme pdc1 containing four cysteines/subunit (positions 69, 152, 221, and 222) with that of a fusion enzyme (pdc1-6, a result of spontaneous gene fusion between PDC1 and PDC6 genes) that is 84% identical in sequence with pdc1 and has only Cys221 (the other three Cys being replaced by aliphatic side chains). The two forms of the enzyme are rather similar so far as steady-state kinetic parameters and substrate activation are considered, as tested for activation by the substrate surrogate pyruvamide. Therefore, if a cysteine is responsible for substrate activation, it must be Cys221. The inactivation of the two enzymes was tested with several inhibitors. Methylmethanethiol sulfonate, a broad spectrum sulfhydryl reagent, could substantially inactivate both enzymes, but was slightly less effective toward the fusion enzyme. (p-Nitrobenzoyl)formic acid is an excellent alternate substrate, whose decarboxylation product p-nitrobenzaldehyde inhibited both enzymes possibly at a Cys221, the only one still present in the fusion enzyme. Exposure of the fusion enzyme, just as of pdc1, to (E)-2-oxo-4-phenyl-3-butenoic acid type inhibitors/alternate substrates enabled detection of the enzyme-bound enamine intermediate at 440 nm. However, unlike pdc1, the fusion enzyme was not irreversibly inactivated by these substrates. These substrates are now known to cause inactivation of pdc1 with concomitant modification of one Cys of the four [Zeng, X.; Chung, A.; Haran, M.; Jordan, F. (1991) J. Am. Chem. Soc. 113, 5842-49].(ABSTRACT TRUNCATED AT 250 WORDS)

  1. Mechanism of Inactivation in Voltage-Gated Na(+) Channels.

    PubMed

    Gawali, V S; Todt, H

    2016-01-01

    Voltage-gated Na(+) channels (VGSCs) initiate action potentials thereby giving rise to rapid transmission of electrical signals along cell membranes and between cells. Depolarization of the cell membrane causes VGSCs to open but also gives rise to a nonconducting state termed inactivation. Inactivation of VGSCs serves a critical physiologic function as it determines the extent of excitability of neurons and of muscle cells. Depending on the time course of development and removal of inactivation both "fast-" and "slow"-inactivated states have been described. Evidence from mutagenesis studies suggests that fast inactivation is produced by a block of the internal vestibule by a tethered inactivation particle that has been mapped to the internal linker between domains III and IV. The motion of this linker may be regulated by parts of the internal C-terminus. The molecular mechanism of slow inactivation is less clear. However, aside from a high number of mutagenesis studies, the recent availability of 3D structures of crystallized prokaryotic VGSCs offers insights into the molecular motions associated with slow inactivation. One possible scenario is that slow movements of the voltage sensors are transmitted to the external vestibule giving rise to a conformational change of this region. This molecular rearrangement is transmitted to the S6 segments giving rise to collapse of the internal vestibule. PMID:27586291

  2. High Pressure Inactivation of Food-borne Viruses

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Over the past half dozen years or so, the USDA Seafood Safety laboratory has endeavored to evaluate the potential of high pressure processing (HPP) for inactivation of food-borne viruses. As a commercial food technology, high pressure processing is highly advantageous because it can inactivate path...

  3. Mechanisms of Escherichia coli inactivation by several disinfectants.

    PubMed

    Cho, Min; Kim, Jaeeun; Kim, Jee Yeon; Yoon, Jeyong; Kim, Jae-Hong

    2010-06-01

    The objective of this study was to elucidate dominant mechanisms of inactivation, i.e. surface attack versus intracellular attack, during application of common water disinfectants such as ozone, chlorine dioxide, free chlorine and UV irradiation. Escherichia coli was used as a representative microorganism. During cell inactivation, protein release, lipid peroxidation, cell permeability change, damage in intracellular enzyme and morphological change were comparatively examined. For the same level of cell inactivation by chemical disinfectants, cell surface damage was more pronounced with strong oxidant such as ozone while damage in inner cell components was more apparent with weaker oxidant such as free chlorine. Chlorine dioxide showed the inactivation mechanism between these two disinfectants. The results suggest that the mechanism of cell inactivation is primarily related to the reactivity of chemical disinfectant. In contrast to chemical disinfectants, cell inactivation by UV occurred without any changes measurable with the methods employed. Understanding the differences in inactivation mechanisms presented herein is critical to identify rate-limiting steps involved in the inactivation process as well as to develop more effective disinfection strategies.

  4. Modeling state-dependent inactivation of membrane currents.

    PubMed Central

    Marom, S; Abbott, L F

    1994-01-01

    Inactivation of many ion channels occurs through largely voltage-independent transitions to an inactivated state from the open state or from other states in the pathway leading to opening of the channel. Because this form of inactivation is state-dependent rather than voltage-dependent, it cannot be described by the standard Hodgkin-Huxley formalism used in virtually all modeling studies of neuronal behavior. Using two examples, cumulative inactivation of the Kv3 potassium channel and inactivation of the fast sodium channel, we extend the standard formalism for modeling macroscopic membrane currents to account for state-dependent inactivation. Our results provide an accurate description of cumulative inactivation of the Kv3 channel, new insight into inactivation of the sodium channel, and a general framework for modeling macroscopic currents when state-dependent processes are involved. In a model neuron, the macroscopic Kv3 current produces a novel short-term memory effect and firing delays similar to those seen in hippocampal neurons. Images FIGURE 5 PMID:7524708

  5. Controlled Inactivation of Recombinant Viruses with Vitamin B2

    PubMed Central

    Callahan, Shellie M.; Wonganan, Piyanuch; Obenauer-Kutner, Linda J.; Sutjipto, Suganto; Dekker, Joseph D.; Croyle, Maria A.

    2008-01-01

    Inactivated viruses are important tools for vaccine development and gene transfer. 8-methoxypsoralen (8-MOP) and long-wavelength ultraviolet irradiation (LWUVI) inactivates many viruses. Toxicity limits its use in animals and humans. Toxicological and photosensitizing properties of riboflavin make it suitable for virus inactivation in preparations for biological use. Viruses expressing beta-galactosidase were mixed with either 8-MOP (1.5 mM) or riboflavin (50 μM) and exposed to LWUVI (365 nm) for 2 hours. Virus activity was determined by limiting dilution. The half-life of the adenovirus preparation treated with 8-MOP was 8.28 nanoseconds−1 (ns−1) and 36.5 ns−1 after treatment with riboflavin. Despite the difference in half-life, both preparations were completely inactivated within 45 minutes. In contrast, the half-lives for adeno-associated virus (AAV) preparations were similar (63 ns−1 8-MOP vs. 67 ns−1 riboflavin). Each AAV preparation was fully inactivated within 90 minutes. The half-life of lentivirus was 193.4 ns−1 after treatment with 8-MOP and 208 ns−1 after exposure to riboflavin. Virus treated with riboflavin was inactivated within 20 minutes. Virus exposed to 8-MOP was inactivated in 90 minutes. DNA and RNA viruses can be inactivated by riboflavin and LWUVI and used in physiological systems sensitive to other photochemicals. PMID:18160141

  6. Kinetics of irreversible inhibition of yeast alcohol dehydrogenase during modification by 4,4'-dithiodipyridine.

    PubMed

    Zheng, S Y; Xu, D; Wang, H R; Li, J; Zhou, H M

    1997-07-01

    The course of inactivation of yeast alcohol dehydrogenase (YADH) using 4,4'-dithiodipyridine (DSDP) has been studied in this paper. The results show that the reaction mechanism between DSDP and YADH is a competitive, complexing inhibition. The microscopic constants for the inactivation of the free enzyme and the enzyme-substrate complex were determined. The presence of the substrate NAD+ offers strong protection for this enzyme against inactivation by DSDP. The above results suggest that two Cys residues are essential for activity and are situated at the active site. These essential Cys residues should be Cys-46 and Cys-174 which are ligands to the catalytic zinc ion. Another Cys residue, which can be modified by DSDP, is non-essential for activity of the enzyme.

  7. Predicting microbial heat inactivation under nonisothermal treatments.

    PubMed

    Hassani, Mounir; Condón, Santiago; Pagán, Rafael

    2007-06-01

    The aim of this study was to develop an equation that accurately predicts microbial heat inactivation under nonisothermal treatments at constantly rising heating rates (from 0.5 to 5 degrees C/min) in media with different pH values (4.0 or 7.4). The survival curves of all bacteria (Enterococcus faecium, Escherichia coli, Listeria monocytogenes, Salmonella Senftenberg 775W, Salmonella Typhimurium, and Staphylococcus aureus) tested under isothermal treatments were nearly linear. For the most heat-resistant microorganism (E. faecium), the estimated DT-values at pH 7.4 were at least 100 times those of the second most thermotolerant microorganism (Salmonella Senftenberg 775W). The heat resistance of E. faecium was up to 30 times lower at pH 4.0 than at pH 7.4. However, E. faecium was still the most heat-resistant microorganism under nonisothermal treatments at both pH values. Inactivation under nonisothermal conditions was not accurately estimated from heat resistance parameters of isothermal treatments when microbial adaptation or sensibilization occurred during the heating up lag phases. The under-prediction of the number of survivors might be greater than 15 log CFU within the nonisothermal treatment conditions investigated. Therefore, the nonisothermal survival curves of the most heat-resistant microorganisms were fitted with the following equation: log S(t) = -(t/delta)P. This equation accurately described the survival curves of all the bacteria tested. We observed a linear relationship between the log of the scale parameter (delta) and the log of the heating rate. A p value characteristic of each microorganism and pH tested was calculated. Two equations capable of predicting the inactivation rate of all bacteria tested under nonisothermal treatments at pH 7.4, 5.5, or 4.0 were developed. The model was evaluated in skim milk and apple juice. The results of this study could be used to help minimize public health risks and to extend the shelf life of those foods

  8. Inactivation and injury of total coliform bacteria after primary disinfection of drinking water by TiO2 photocatalysis.

    PubMed

    Rizzo, Luigi

    2009-06-15

    In this study the potential application of TiO(2) photocatalysis as primary disinfection system of drinking water was investigated in terms of coliform bacteria inactivation and injury. As model water the effluent of biological denitrification unit for nitrate removal from groundwater, which is characterized by high organic matter and bacteria release, was used. The injury of photocatalysis on coliform bacteria was characterized by means of selective (mEndo) and less selective (mT7) culture media. Different catalyst loadings as well as photolysis and adsorption effects were investigated. Photocatalysis was effective in coliform bacteria inactivation (91-99% after 60 min irradiation time, depending on both catalyst loading and initial density of coliform bacteria detected by mEndo), although no total removal was observed after 60 min irradiation time. The contribution of adsorption mechanism was significant (60-98% after 60 min, depending on catalyst loading) compared to previous investigations probably due to the nature of source water rich in particulate organic matter and biofilm. Photocatalysis process did not result in any irreversible injury (98.8% being the higher injury) under investigated conditions, thus a bacteria regrowth may take place under optimum environment conditions if any final disinfection process (e.g., chlorine or chlorine dioxide) is not used.

  9. hSNF5/INI1 inactivation is mainly associated with homozygous deletions and mitotic recombinations in rhabdoid tumors.

    PubMed

    Rousseau-Merck, M F; Versteege, I; Legrand, I; Couturier, J; Mairal, A; Delattre, O; Aurias, A

    1999-07-01

    The chromatin-remodeling hSNF5/INI1 gene has recently been shown to act as a tumor suppressor gene in rhabdoid tumors (RTs). In an attempt to further characterize the main chromosomal mechanisms involved in hSNF5/INI1 inactivation in RTs, we report here the molecular cytogenetic data obtained in 12 cell lines harboring hSNF5/INI1 mutations and/or deletions in relation to the molecular genetic analysis using polymorphic markers extended to both extremities of chromosome 22q. On the whole, mitotic recombination occurring in the proximal part of chromosome 22q, as demonstrated in five cases, and nondisjunction/duplication, highly suspected in two cases (processes leading respectively to partial or complete isodisomy), appear to be major mechanisms associated with hSNF5/INI1 inactivation. Such isodisomy accompanies each of the RTs exhibiting two cytogenetically normal chromosomes 22. This results in homozygosity for the mutation at the hSNF5/INI1 locus. An alternate mechanism accounting for hSNF5/INI1 inactivation observed in these tumors is homozygous deletion in the rhabdoid consensus region. This was observed in each of the four tumors carrying a chromosome 22q abnormality and, in particular, in the three tumors with chromosomal translocations. Only one case of our series illustrates the mutation/deletion classical model proposed for the double-hit inactivation of a tumor suppressor gene. PMID:10397258

  10. Irreversible Heating Measurement with Microsecond Pulse Magnet: Example of the α-θ Phase Transition of Solid Oxygen

    NASA Astrophysics Data System (ADS)

    Nomura, Toshihiro; Matsuda, Yasuhiro H.; Takeyama, Shojiro; Kobayashi, Tatsuo C.

    2016-09-01

    Dissipation inevitably occurs in first-order phase transitions, leading to irreversible heating. Conversely, the irreversible heating effect may indicate the occurrence of the first-order phase transition. We measured the temperature change at the magnetic-field-induced α-θ phase transition of solid oxygen. A significant temperature increase from 13 to 37 K, amounting to 700 J/mol, due to irreversible heating was observed at the first-order phase transition. We argue that the hysteresis loss of the magnetization curve and the dissipative structural transformation account for the irreversible heating. The measurement of irreversible heating can be utilized to detect the first-order phase transition in combination with an ultrahigh magnetic fields generated in a time of µs order.

  11. Antibacterial efficacy and effect of Morinda citrifolia L. mixed with irreversible hydrocolloid for dental impressions: A randomized controlled trial

    PubMed Central

    Ahmed, A. Shafath; Charles, P. David; Cholan, R.; Russia, M.; Surya, R.; Jailance, L.

    2015-01-01

    Aim: This study aimed to evaluate whether the extract of Morinda citrifolia L. mixed with irreversible hydrocolloid powder decreases microbial contamination during impression making without affecting the resulting casts. Materials and Methods: Twenty volunteers were randomly divided into two groups (n = 10). Group A 30 ml extract of M. citrifolia L diluted in 30 ml of water was mixed to make the impression with irreversible hydrocolloid material. Group B 30 ml deionized water was mixed with irreversible hydrocolloid material to make the impressions following which the surface roughness and dimensional stability of casts were evaluated. Results: Extract of M. citrifolia L. mixed with irreversible hydrocolloid decreased the percentage of microorganisms when compared with water (P < 0.001) but did not affect the surface quality or dimensional stability of the casts. Conclusion: Mixing the extract of M. citrifolia L. with irreversible hydrocolloid powder is an alternative method to prevent contamination without sacrificing impression quality. PMID:26538926

  12. X-inactivation patterns in monozygotic and dizygotic female twins

    SciTech Connect

    Goodship, J.; Carter, J.; Burn, J.

    1996-01-22

    We have tested the hypothesis that contrasting X-inactivation patterns could be a trigger for monozygotic twinning in females. X-inactivation patterns were studied in umbilical cord tissue in 43 monozygotic twin pairs and 24 dizygotic twin pairs. Very skewed or non-random X-inactivation patterns were observed in both twins in six of the monozygotic twin pairs and in one of the dizygotic twin pairs. Contrasting X-inactivation patterns occurred in only one of the six monozygotic twin pairs. This does not support the original hypothesis. There is a trend to extreme skewing of X-inactivation pattern occurring more frequently in monozygotic twins. 21 refs., 1 fig., 2 tabs.

  13. Chlorophyll mediated photodynamic inactivation of blue laser on Streptococcus mutans

    NASA Astrophysics Data System (ADS)

    Astuti, Suryani Dyah; Zaidan, A.; Setiawati, Ernie Maduratna; Suhariningsih

    2016-03-01

    Photodynamic inactivation is an inactivation method in microbial pathogens that utilize light and photosensitizer. This study was conducted to investigate photodynamic inactivation effects of low intensity laser exposure with various dose energy on Streptococcus mutans bacteria. The photodynamic inactivation was achieved with the addition of chlorophyll as photosensitizers. To determine the survival percentage of Streptococcus mutans bacteria after laser exposure, the total plate count method was used. For this study, the wavelength of the laser is 405 nm and variables of energy doses are 1.44, 2.87, 4.31, 5.74, 7.18, and 8.61 in J/cm2. The results show that exposure to laser with energy dose of 7.18 J/cm2 has the best photodynamic inactivation with a decrease of 78% in Streptococcus

  14. [Inactivation of T4 phage in water environment using proteinase].

    PubMed

    Lü, Wen-zhou; Yang, Qing-xiang; Zhang, Yu; Yang, Min; Zhu, Chun-fang

    2004-09-01

    The inactivation effectiveness of proteinase to viruses was investigated by using T4 phage as a model virus. The results showed that the inactivation effectiveness of proteinase to T4 phage was obvious. In the optimum conditions and 67.5 u/mL concentration, the inactivation rate of proteinase K to T4 phage in sterilized water and in sewage achieved 99.4% and 49.4% respectively in an hour, and achieved >99.9% and 81.1% in three hours. The inactivation rate of the industrial proteinase 1398 to T4 phage in sterilized water achieved 74.4% in an hour. The effects of pH and temperature on the inactivation effectiveness was not evident.

  15. Partially integrated exhaust manifold

    SciTech Connect

    Hayman, Alan W; Baker, Rodney E

    2015-01-20

    A partially integrated manifold assembly is disclosed which improves performance, reduces cost and provides efficient packaging of engine components. The partially integrated manifold assembly includes a first leg extending from a first port and terminating at a mounting flange for an exhaust gas control valve. Multiple additional legs (depending on the total number of cylinders) are integrally formed with the cylinder head assembly and extend from the ports of the associated cylinder and terminate at an exit port flange. These additional legs are longer than the first leg such that the exit port flange is spaced apart from the mounting flange. This configuration provides increased packaging space adjacent the first leg for any valving that may be required to control the direction and destination of exhaust flow in recirculation to an EGR valve or downstream to a catalytic converter.

  16. Partially coherent ultrafast spectrography

    PubMed Central

    Bourassin-Bouchet, C.; Couprie, M.-E.

    2015-01-01

    Modern ultrafast metrology relies on the postulate that the pulse to be measured is fully coherent, that is, that it can be completely described by its spectrum and spectral phase. However, synthesizing fully coherent pulses is not always possible in practice, especially in the domain of emerging ultrashort X-ray sources where temporal metrology is strongly needed. Here we demonstrate how frequency-resolved optical gating (FROG), the first and one of the most widespread techniques for pulse characterization, can be adapted to measure partially coherent pulses even down to the attosecond timescale. No modification of experimental apparatuses is required; only the processing of the measurement changes. To do so, we take our inspiration from other branches of physics where partial coherence is routinely dealt with, such as quantum optics and coherent diffractive imaging. This will have important and immediate applications, such as enabling the measurement of X-ray free-electron laser pulses despite timing jitter. PMID:25744080

  17. Laparoscopic partial splenic resection.

    PubMed

    Uranüs, S; Pfeifer, J; Schauer, C; Kronberger, L; Rabl, H; Ranftl, G; Hauser, H; Bahadori, K

    1995-04-01

    Twenty domestic pigs with an average weight of 30 kg were subjected to laparoscopic partial splenic resection with the aim of determining the feasibility, reliability, and safety of this procedure. Unlike the human spleen, the pig spleen is perpendicular to the body's long axis, and it is long and slender. The parenchyma was severed through the middle third, where the organ is thickest. An 18-mm trocar with a 60-mm Endopath linear cutter was used for the resection. The tissue was removed with a 33-mm trocar. The operation was successfully concluded in all animals. No capsule tears occurred as a result of applying the stapler. Optimal hemostasis was achieved on the resected edges in all animals. Although these findings cannot be extended to human surgery without reservations, we suggest that diagnostic partial resection and minor cyst resections are ideal initial indications for this minimally invasive approach.

  18. Partially coherent ultrafast spectrography

    NASA Astrophysics Data System (ADS)

    Bourassin-Bouchet, C.; Couprie, M.-E.

    2015-03-01

    Modern ultrafast metrology relies on the postulate that the pulse to be measured is fully coherent, that is, that it can be completely described by its spectrum and spectral phase. However, synthesizing fully coherent pulses is not always possible in practice, especially in the domain of emerging ultrashort X-ray sources where temporal metrology is strongly needed. Here we demonstrate how frequency-resolved optical gating (FROG), the first and one of the most widespread techniques for pulse characterization, can be adapted to measure partially coherent pulses even down to the attosecond timescale. No modification of experimental apparatuses is required; only the processing of the measurement changes. To do so, we take our inspiration from other branches of physics where partial coherence is routinely dealt with, such as quantum optics and coherent diffractive imaging. This will have important and immediate applications, such as enabling the measurement of X-ray free-electron laser pulses despite timing jitter.

  19. Sucrose density gradient centrifugation and cross-flow filtration methods for the production of arbovirus antigens inactivated by binary ethylenimine

    PubMed Central

    Pyke, Alyssa T; Phillips, Debra A; Chuan, Teck F; Smith, Greg A

    2004-01-01

    Background Sucrose density gradient centrifugation and cross-flow filtration methods have been developed and standardised for the safe and reproducible production of inactivated arbovirus antigens which are appropriate for use in diagnostic serological applications. Methods To optimise the maximum titre of growth during the propagation of arboviruses, the multiplicity of infection and choice of cell line were investigated using stocks of Ross River virus and Barmah Forest virus grown in both mosquito and mammalian cell lines. To standardise and improve the efficacy of the inactivation of arboviral suspensions, stocks of Ross River virus, Barmah Forest virus, Japanese encephalitis virus, Murray Valley encephalitis virus and Alfuy virus were chemically inactivated using binary ethylenimine at a final concentration of 3 mM. Aliquots were then taken at hourly intervals and crude inactivation rates were determined for each virus using a plaque assay. To ensure complete inactivation, the same aliquots were each passaged 3 times in Aedes albopictus C6/36 cells and the presence of viral growth was detected using an immunofluorescent assay. For larger quantities of viral suspensions, centrifugation on an isopycnic sucrose density gradient or cross-flow filtration was used to produce concentrated, pure antigens or partially concentrated, semi-purified antigens respectively. Results The results of the propagation experiments suggested that the maximum viral titres obtained for both Ross River virus and Barmah Forest virus were affected by the incubation period and choice of cell line, rather than the use of different multiplicity of infection values. Results of the binary ethylenimine inactivation trial suggested that standardised periods of 5 or 8 hours would be suitable to ensure effective and complete inactivation for a number of different arboviral antigens. Conclusion Two methods used to prepare inactivated arbovirus antigens have been standardised to minimise production

  20. Thermal inactivation of ricin using infant formula as a food matrix.

    PubMed

    Jackson, Lauren S; Tolleson, William H; Chirtel, Stuart J

    2006-09-20

    Ricin is a potent protein toxin found in the seeds of the castor bean plant, Ricinus communis. Ricin specifically and irreversibly inactivates ribosomes, promoting cell death by inhibiting protein synthesis. It is composed of a ribosome-inactivating enzyme (A-chain) linked to a lectin (B-chain) by a single disulfide bond. Several reports indicate that ricin can be detoxified by thermal treatment; however, the conditions required for inactivation are not well characterized. In addition, little information exists on the thermal stability of ricin added to foods. The objective of this work was to determine the effects of heat treatments on the detection and toxicity of ricin added to milk- and soy-based infant formulas. Reconstituted infant formula powders containing 100 mug of ricin/mL were heated at 60-90 degrees C for up to 5 h. The heat-treated formulas were analyzed by ELISA to determine levels of ricin. The residual cytotoxicity of ricin-containing infant formula after heat treatments was determined using RAW264.7 mouse macrophage cells. The ELISA and the cytotoxicity assay indicated that ricin detection and toxicity decreased with increasing heating times and temperatures. Minimal losses in detection and toxicity were found for ricin heated at 60 degrees C for 2 h. The half-lives of ricin cytoxic activity in a milk-based infant formula at 60, 70, 75, 80, 85, and 90 degrees C were >100, 9.8 +/- 0.5, 5.8 +/- 0.9, 5.1 +/- 0.7, 3.1 +/- 0.4, and 1.8 +/- 0.2 min, respectively; the comparable values for a soy-based infant formula were >100, 16 +/- 1.6, 8.7 +/- 1.2, 6.9 +/- 1.1, 3.0 +/- 0.4, and 2.0 +/- 0.3 min. ELISA detection was a good indicator of the cytotoxicity of heat-treated ricin. The results indicate that ricin is a relatively heat stable protein and may remain toxic under some food processing conditions.

  1. Stimulation and oxidative catalytic inactivation of thermolysin by copper.Cys-Gly-His-Lys.

    PubMed

    Gokhale, Nikhil H; Bradford, Seth; Cowan, J A

    2007-09-01

    [Cu(2+).Cys-Gly-His-Lys] stimulates thermolysin (TLN) activity at low concentration (below 10 microM) and inhibits the enzyme at higher concentration, with binding affinities of 2.0 and 4.9 microM, respectively. The metal-free Cys-Gly-His-Lys peptide also stimulates TLN activity, with an apparent binding affinity of 2.2 microM. Coordination of copper through deprotonated imine nitrogens, the histidyl nitrogen, and the free N-terminal amino group is consistent with the characteristic absorption spectrum of a Cu(2+)-amino-terminal copper and nickel binding motif (lambda (max) approximately 525 nm). The lack of thiol coordination is suggested by both the absence of a thiol to Cu(2+) charge transfer band and electrochemical studies, since the electrode potential (vs. Ag/AgCl) 0.84 V (DeltaE = 92 mV) for the Cu(3+/2+) redox couple obtained for [Cu(2+).Cys-Gly-His-Lys] was found to be in close agreement with that of a related complex [Cu(2+).Lys-Gly-His-Lys](+) (0.84 V, DeltaE = 114 mV). The N-terminal cysteine appears to be available as a zinc-anchoring residue and plays a critical functional role since the [Cu(2+).Lys-Gly-His-Lys](+) homologue exhibits neither stimulation nor inhibition of TLN. Under oxidizing conditions (ascorbate/O(2)) the catalyst is shown to mediate the complete irreversible inactivation of TLN at concentrations where enzyme activity would otherwise be stimulated. The observed rate constant for inactivation of TLN activity was determined as k (obs) = 7.7 x 10(-2) min(-1), yielding a second-order rate constant of (7.7 +/- 0.9) x 10(4) M(-1) min(-1). Copper peptide mediated generation of reactive oxygen species that subsequently modify active-site residues is the most likely pathway for inactivation of TLN rather than cleavage of the peptide backbone. PMID:17618468

  2. Inactivation of the Lactobacillus leichmannii ribonucleoside triphosphate reductase by 2'-chloro-2'-deoxyuridine 5'-triphosphate: stoichiometry of inactivation, site of inactivation, and mechanism of the protein chromophore formation

    SciTech Connect

    Ashley, G.W.; Harris, G.; Stubbe, J.A.

    1988-06-14

    The ribonucleoside triphosphate reductase (RTPR) of Lactobacillus leichmannii is inactivated by the substrate analogue 2'-chloro-2'-deoxyuridine 5'-triphosphate (ClUTP). Inactivation is due to alkylation by 2-methylene-3(2H)-furanone, a decomposition product of the enzymic product 3'-keto-2'-deoxyuridine triphosphate. The former has been unambiguously identified as 2-((ethylthio)methyl)-3(2H)-furanone, an ethanethiol trapped adduct, which is identical by /sup 1/H NMR spectroscopy with material synthesized chemically. Subsequent to rapid inactivation, a slow process occurs that results in formation of a new protein-associated chromophore absorbing maximally near 320 nm. The terminal stages of the inactivation have now been investigated in detail. The alkylation and inactivation stoichiometries were studied as a function of the ratio of ClUTP to enzyme. The amount of labeling of RTPR increased with increasing ClUTP concentration up to the maximum of approximately 4 labels/RTPR, yet the degree of inactivation did not increase proportionally. This suggests that (1) RTPR may be inactivated by alkylation of a single site and (2) decomposition of 3'-keto-dUTP is not necessarily enzyme catalyzed. The formation of the new protein chromophore was also monitored during inactivation and found to reach its full extent upon the first alkylation . Thus, out of four alkylation sites, only one appears capable of undergoing the subsequent reaction to form the new chromophore. Model studies suggest that the new chromophore is due to addition of an amino group to the 5-position of enzyme-bound furanone, followed by ring opening and tautomerization to give a ..beta..-aminoenone structure.

  3. Inactivation of microbes using ultrasound: a review.

    PubMed

    Piyasena, P; Mohareb, E; McKellar, R C

    2003-11-01

    Alternative methods for pasteurization and sterilization are gaining importance, due to increased consumer demand for new methods of food processing that have a reduced impact on nutritional content and overall food quality. Ultrasound processing or sonication is one of the alternative technologies that has shown promise in the food industry. Sonication alone is not very effective in killing bacteria in food; however, the use of ultrasound coupled with pressure and/or heat is promising. Thermosonic (heat plus sonication), manosonic (pressure plus sonication), and manothermosonic (heat and pressure plus sonication) treatments are likely the best methods to inactivate microbes, as they are more energy-efficient and effective in killing microorganisms. Ultrasonic processing is still in its infancy and requires a great deal of future research in order to develop the technology on an industrial scale, and to more fully elucidate the effect of ultrasound on the properties of foods.

  4. UV inactivation of pathogenic and indicator microorganisms

    SciTech Connect

    Chang, J.C.; Ossoff, S.F.; Lobe, D.C.; Dorfman, M.H.; Dumais, C.M.; Qualls, R.G.; Johnson, J.D.

    1985-06-01

    Survival was measured as a function of the dose of germicidal UV light for the bacteria Escherichia coli, Salmonella typhi, Shigella sonnei, Streptococcus faecalis, Staphylococcus aureus, and Bacillus subtilis spores, the enteric viruses poliovirus type 1 and simian rotavirus SA11, the cysts of the protozoan Acanthamoeba castellanii, as well as for total coliforms and standard plate count microorganisms from secondary effluent. The doses of UV light necessary for a 99.9% inactivation of the cultured vegetative bacteria, total coliforms, and standard plate count microorganisms were comparable. However, the viruses, the bacterial spores, and the amoebic cysts required about 3 to 4 times, 9 times, and 15 times, respectively, the dose required for E. coli. These ratios covered a narrower relative dose range than that previously reported for chlorine disinfection of E. coli, viruses, spores, and cysts.

  5. Resistance in antimicrobial photodynamic inactivation of bacteria.

    PubMed

    Maisch, Tim

    2015-08-01

    Antibiotics have increasingly lost their impact to kill bacteria efficiently during the last 10 years. The emergence and dissemination of superbugs with resistance to multiple antibiotic classes have occurred among Gram-positive and Gram-negative strains including Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacter strains. These six superbugs can "escape" more or less any single kind of antibiotic treatment. That means bacteria are very good at developing resistance against antibiotics in a short time. One new approach is called photodynamic antimicrobial chemotherapy (PACT) which already has demonstrated an efficient antimicrobial efficacy among multi-resistant bacteria. Until now it has been questionable if bacteria can develop resistance against PACT. This perspective summarises the current knowledge about the susceptibility of bacteria towards oxidative stress and sheds some light on possible strategies of the development of photodynamic inactivation of bacteria (PACT)-induced oxidative stress resistance by bacteria.

  6. Streptomyces erythraeus Trypsin Inactivates α1-Antitrypsin

    PubMed Central

    Vukoti, Krishna M.; Kadiyala, Chandra Sekhar Rao; Miyagi, Masaru

    2011-01-01

    Streptomyces erythraeus trypsin (SET) is a serine protease that is secreted extracellularly by Streptomyces erythraeus. We investigated the inhibitory effect of α1-antitrypsin on the catalytic activity of SET. Intriguingly, we found that SET is not inhibited by α1-antitrypsin. Our investigations into the molecular mechanism underlying this observation revealed that SET hydrolyzes the Met-Ser bond in the reaction center loop of α1-antitrypsin. However, SET somehow avoids entrapment by α1-antitrypsin. We also confirmed that α1-antitrypsin loses its inhibitory activity after incubation with SET. Thus, our study demonstrates that SET is not only resistant to α1-antitrypsin but also inactivates α1-antitrypsin. PMID:22115549

  7. Ribosome-Inactivating and Related Proteins

    PubMed Central

    Schrot, Joachim; Weng, Alexander; Melzig, Matthias F.

    2015-01-01

    Ribosome-inactivating proteins (RIPs) are toxins that act as N-glycosidases (EC 3.2.2.22). They are mainly produced by plants and classified as type 1 RIPs and type 2 RIPs. There are also RIPs and RIP related proteins that cannot be grouped into the classical type 1 and type 2 RIPs because of their different sizes, structures or functions. In addition, there is still not a uniform nomenclature or classification existing for RIPs. In this review, we give the current status of all known plant RIPs and we make a suggestion about how to unify those RIPs and RIP related proteins that cannot be classified as type 1 or type 2 RIPs. PMID:26008228

  8. Approaches to Inactivate Genes in Zebrafish.

    PubMed

    Parant, John M; Yeh, Jing-Ruey Joanna

    2016-01-01

    Animal models of tumor initiation and tumor progression are essential components toward understanding cancer and designing/validating future therapies. Zebrafish is a powerful model for studying tumorigenesis and has been successfully exploited in drug discovery. According to the zebrafish reference genome, 82 % of disease-associated genes in the Online Mendelian Inheritance in Man (OMIM) database have clear zebrafish orthologues. Using a variety of large-scale random mutagenesis methods developed to date, zebrafish can provide a unique opportunity to identify gene mutations that may be associated with cancer predisposition. On the other hand, newer technologies enabling targeted mutagenesis can facilitate reverse cancer genetic studies and open the door for complex genetic analysis of tumorigenesis. In this chapter, we will describe the various technologies for conducting genome editing in zebrafish with special emphasis on the approaches to inactivate genes. PMID:27165349

  9. UV inactivation of pathogenic and indicator microorganisms.

    PubMed Central

    Chang, J C; Ossoff, S F; Lobe, D C; Dorfman, M H; Dumais, C M; Qualls, R G; Johnson, J D

    1985-01-01

    Survival was measured as a function of the dose of germicidal UV light for the bacteria Escherichia coli, Salmonella typhi, Shigella sonnei, Streptococcus faecalis, Staphylococcus aureus, and Bacillus subtilis spores, the enteric viruses poliovirus type 1 and simian rotavirus SA11, the cysts of the protozoan Acanthamoeba castellanii, as well as for total coliforms and standard plate count microorganisms from secondary effluent. The doses of UV light necessary for a 99.9% inactivation of the cultured vegetative bacteria, total coliforms, and standard plate count microorganisms were comparable. However, the viruses, the bacterial spores, and the amoebic cysts required about 3 to 4 times, 9 times, and 15 times, respectively, the dose required for E. coli. These ratios covered a narrower relative dose range than that previously reported for chlorine disinfection of E. coli, viruses, spores, and cysts. PMID:2990336

  10. Inactivation of Coxiella burnetti by gamma irradiation

    SciTech Connect

    Scott, G.H.; McCaul, T.F.; Williams, J.C.

    1989-01-01

    The gamma radiation inactivation kinetics for Coxiella burnetii at - 79 C were exponential. The radiation dose needed to reduce the number of infective C. burnetii by 90% varied from 0-64 to 1.2 kGy depending on the phase of hte micro-organism, purity of the culture and composition of suspending menstruum. The viability of preparations containing C. burnetti was completely abolished by 10 kGy without diminishing antigenicity or ability to elicit a protective immune response in vaccinated mice. Immunocytochemical examinations using monoclonal antibodies and electron microscopy demonstrated that radiation doses of 20 kGy did not alter cell-wall morphology or cell-surface antigenic epitopes.

  11. Assessing the role of feed water constituents in irreversible membrane fouling of pilot-scale ultrafiltration drinking water treatment systems.

    PubMed

    Peiris, R H; Jaklewicz, M; Budman, H; Legge, R L; Moresoli, C

    2013-06-15

    Fluorescence excitation-emission matrix (EEM) approach together with principal component analysis (PCA) was used for assessing hydraulically irreversible fouling of three pilot-scale ultrafiltration (UF) systems containing full-scale and bench-scale hollow fiber membrane modules in drinking water treatment. These systems were operated for at least three months with extensive cycles of permeation, combination of back-pulsing and scouring and chemical cleaning. The principal component (PC) scores generated from the PCA of the fluorescence EEMs were found to be related to humic substances (HS), protein-like and colloidal/particulate matter content. PC scores of HS- and protein-like matter of the UF feed water, when considered separately, showed reasonably good correlations with the rate of hydraulically irreversible fouling for long-term UF operations. In contrast, comparatively weaker correlations for PC scores of colloidal/particulate matter and the rate of hydraulically irreversible fouling were obtained for all UF systems. Since, individual correlations could not fully explain the evolution of the rate of irreversible fouling, multi-linear regression models were developed to relate the combined effect of HS-like, protein-like and colloidal/particulate matter PC scores to the rate of hydraulically irreversible fouling for each specific UF system. These multi-linear regression models revealed significant individual and combined contribution of HS- and protein-like matter to the rate of hydraulically irreversible fouling, with protein-like matter generally showing the greatest contribution. The contribution of colloidal/particulate matter to the rate of hydraulically irreversible fouling was not as significant. The addition of polyaluminum chloride, as coagulant, to UF feed appeared to have a positive impact in reducing hydraulically irreversible fouling by these constituents. The proposed approach has applications in quantifying the individual and synergistic

  12. Assessing the role of feed water constituents in irreversible membrane fouling of pilot-scale ultrafiltration drinking water treatment systems.

    PubMed

    Peiris, R H; Jaklewicz, M; Budman, H; Legge, R L; Moresoli, C

    2013-06-15

    Fluorescence excitation-emission matrix (EEM) approach together with principal component analysis (PCA) was used for assessing hydraulically irreversible fouling of three pilot-scale ultrafiltration (UF) systems containing full-scale and bench-scale hollow fiber membrane modules in drinking water treatment. These systems were operated for at least three months with extensive cycles of permeation, combination of back-pulsing and scouring and chemical cleaning. The principal component (PC) scores generated from the PCA of the fluorescence EEMs were found to be related to humic substances (HS), protein-like and colloidal/particulate matter content. PC scores of HS- and protein-like matter of the UF feed water, when considered separately, showed reasonably good correlations with the rate of hydraulically irreversible fouling for long-term UF operations. In contrast, comparatively weaker correlations for PC scores of colloidal/particulate matter and the rate of hydraulically irreversible fouling were obtained for all UF systems. Since, individual correlations could not fully explain the evolution of the rate of irreversible fouling, multi-linear regression models were developed to relate the combined effect of HS-like, protein-like and colloidal/particulate matter PC scores to the rate of hydraulically irreversible fouling for each specific UF system. These multi-linear regression models revealed significant individual and combined contribution of HS- and protein-like matter to the rate of hydraulically irreversible fouling, with protein-like matter generally showing the greatest contribution. The contribution of colloidal/particulate matter to the rate of hydraulically irreversible fouling was not as significant. The addition of polyaluminum chloride, as coagulant, to UF feed appeared to have a positive impact in reducing hydraulically irreversible fouling by these constituents. The proposed approach has applications in quantifying the individual and synergistic

  13. Regulation of X-chromosome inactivation by the X-inactivation centre.

    PubMed

    Augui, Sandrine; Nora, Elphège P; Heard, Edith

    2011-06-01

    X-chromosome inactivation (XCI) ensures dosage compensation in mammals and is a paradigm for allele-specific gene expression on a chromosome-wide scale. Important insights have been made into the developmental dynamics of this process. Recent studies have identified several cis- and trans-acting factors that regulate the initiation of XCI via the X-inactivation centre. Such studies have shed light on the relationship between XCI and pluripotency. They have also revealed the existence of dosage-dependent activators that trigger XCI when more than one X chromosome is present, as well as possible mechanisms underlying the monoallelic regulation of this process. The recent discovery of the plasticity of the inactive state during early development, or during cloning, and induced pluripotency have also contributed to the X chromosome becoming a gold standard in reprogramming studies.

  14. Sunlight inactivation of viruses in open-water unit process treatment wetlands: modeling endogenous and exogenous inactivation rates.

    PubMed

    Silverman, Andrea I; Nguyen, Mi T; Schilling, Iris E; Wenk, Jannis; Nelson, Kara L

    2015-03-01

    Sunlight inactivation is an important mode of disinfection for viruses in surface waters. In constructed wetlands, for example, open-water cells can be used to promote sunlight disinfection and remove pathogenic viruses from wastewater. To aid in the design of these systems, we developed predictive models of virus attenuation that account for endogenous and exogenous sunlight-mediated inactivation mechanisms. Inactivation rate models were developed for two viruses, MS2 and poliovirus type 3; laboratory- and field-scale experiments were conducted to evaluate the models' ability to estimate inactivation rates in a pilot-scale, open-water, unit-process wetland cell. Endogenous inactivation rates were modeled using either photoaction spectra or total, incident UVB irradiance. Exogenous inactivation rates were modeled on the basis of virus susceptibilities to singlet oxygen. Results from both laboratory- and field-scale experiments showed good agreement between measured and modeled inactivation rates. The modeling approach presented here can be applied to any sunlit surface water and utilizes easily measured inputs such as depth, solar irradiance, water matrix absorbance, singlet oxygen concentration, and the virus-specific apparent second-order rate constant with singlet oxygen (k2). Interestingly, the MS2 k2 in the open-water wetland was found to be significantly larger than k2 observed in other waters in previous studies. Examples of how the model can be used to design and optimize natural treatment systems for virus inactivation are provided.

  15. Mechanism-based inactivation of benzo(a)pyrene hydroxylase by aryl acetylenes and aryl olefins

    SciTech Connect

    Gan, L.S.; Lu, J.Y.L.; Alworth, W.L.

    1986-05-01

    A series of aryl acetylenes and aryl olefins have been examined as substrates and inhibitors of cytochrome P-450 dependent monooxgenases in liver microsomes from 5,6-benzoflavone or phenobarbital pretreated rats. 1-Ethynylpyrene, 3-ethynylperylene, 2-ethynylfluorene, methyl 1-pyrenyl acetylene, cis- and trans-1-(2-bromovinyl)pyrene, and 1-allylpyrene serve as mechanism-based irreversible inactivators (suicide inhibitors) of benzo(a)pyrene hydroxylase, while 1-vinylpyrene and phenyl 1-pyrenyl acetylene do not cause a detectable suicide inhibition of benzo(a)pyrene hydroxylase. The mechanism-based loss of benzo(a)pyrene hydroxylase caused by the aryl acetylenes is not accompanied by a corresponding loss of the P-450 content of the microsomes (suicide destruction). The suicide inhibition by these aryl acetylenes therefore does not involve covalent binding to the heme moiety of the monooxygenase. Nevertheless, in the presence of NADPH, /sup 3/H-labeled 1-ethynylpyrene becomes covalently attached to the cytochrome P-450 protein; the measured stoichiometry of binding is one 1-ethynylpyrene per P-450 heme unit. The authors conclude that the inhibition of benzo(a)pyrene hydroxylase produced by 1-ethynylpyrene may be related to the mechanism of suicide inhibition of P-450 activity by chloramphenicol rather than the mechanism of suicide destruction of P-450 previously described for acetylene and propyne.

  16. A new age for biomedical applications of Ribosome Inactivating Proteins (RIPs): from bioconjugate to nanoconstructs.

    PubMed

    Pizzo, Elio; Di Maro, Antimo

    2016-07-20

    Ribosome-inactivating proteins (RIPs) are enzymes (3.2.2.22) that possess N-glycosilase activity that irreversibly inhibits protein synthesis. RIPs have been found in plants, fungi, algae, and bacteria; their biological role is still under investigation, even if it has been recognized their role in plant defence against predators and viruses. Nevertheless, several studies on these toxins have been performed to evaluate their applicability in the biomedical field making RIPs selectively toxic towards target cells. Indeed, these molecules are extensively used to produce chimeric biomolecules, such as immunotoxins or protein/peptides conjugates. However, to date, clinical use of most of these bioconiujates has been limited by toxicity and immunogenicity. More recently, material sciences have provided a wide range of nanomaterials to be used as excellent vehicles for toxin-delivery, since they are characterized by improved stability, solubility, and in vivo pharmacokinetics. This review discusses progresses in the development of RIPs bioconjugates, with particular attention to the recent use of nanomaterials, whose appropriate design opens up a broad range of different possibilities to the use of RIPs in novel therapeutic approaches in human diseases.

  17. Nitration and Inactivation of Manganese Superoxide Dismutase in Chronic Rejection of Human Renal Allografts

    NASA Astrophysics Data System (ADS)

    MacMillan-Crow, L. A.; Crow, John P.; Kerby, Jeffrey D.; Beckman, Joseph S.; Thompson, John A.

    1996-10-01

    Inflammatory processes in chronic rejection remain a serious clinical problem in organ transplantation. Activated cellular infiltrate produces high levels of both superoxide and nitric oxide. These reactive oxygen species interact to form peroxynitrite, a potent oxidant that can modify proteins to form 3-nitrotyrosine. We identified enhanced immunostaining for nitrotyrosine localized to tubular epithelium of chronically rejected human renal allografts. Western blot analysis of rejected tissue demonstrated that tyrosine nitration was restricted to a few specific polypeptides. Immunoprecipitation and amino acid sequencing techniques identified manganese superoxide dismutase, the major antioxidant enzyme in mitochondria, as one of the targets of tyrosine nitration. Total manganese superoxide dismutase protein was increased in rejected kidney, particularly in the tubular epithelium; however, enzymatic activity was significantly decreased. Exposure of recombinant human manganese superoxide dismutase to peroxynitrite resulted in a dose-dependent (IC50 = 10 μ M) decrease in enzymatic activity and concomitant increase in tyrosine nitration. Collectively, these observations suggest a role for peroxynitrite during development and progression of chronic rejection in human renal allografts. In addition, inactivation of manganese superoxide dismutase by peroxynitrite may represent a general mechanism that progressively increases the production of peroxynitrite, leading to irreversible oxidative injury to mitochondria.

  18. A new age for biomedical applications of Ribosome Inactivating Proteins (RIPs): from bioconjugate to nanoconstructs.

    PubMed

    Pizzo, Elio; Di Maro, Antimo

    2016-01-01

    Ribosome-inactivating proteins (RIPs) are enzymes (3.2.2.22) that possess N-glycosilase activity that irreversibly inhibits protein synthesis. RIPs have been found in plants, fungi, algae, and bacteria; their biological role is still under investigation, even if it has been recognized their role in plant defence against predators and viruses. Nevertheless, several studies on these toxins have been performed to evaluate their applicability in the biomedical field making RIPs selectively toxic towards target cells. Indeed, these molecules are extensively used to produce chimeric biomolecules, such as immunotoxins or protein/peptides conjugates. However, to date, clinical use of most of these bioconiujates has been limited by toxicity and immunogenicity. More recently, material sciences have provided a wide range of nanomaterials to be used as excellent vehicles for toxin-delivery, since they are characterized by improved stability, solubility, and in vivo pharmacokinetics. This review discusses progresses in the development of RIPs bioconjugates, with particular attention to the recent use of nanomaterials, whose appropriate design opens up a broad range of different possibilities to the use of RIPs in novel therapeutic approaches in human diseases. PMID:27439918

  19. Chromophore-Assisted Light Inactivation and Self-Organization of Microtubules and Motors

    NASA Astrophysics Data System (ADS)

    Surrey, Thomas; Elowitz, Michael B.; Wolf, Pierre-Etienne; Yang, Feng; Nedelec, Francois; Shokat, Kevan; Leibler, Stanislas

    1998-04-01

    Chromophore-assisted light inactivation (CALI) offers the only method capable of modulating specific protein activities in localized regions and at particular times. Here, we generalize CALI so that it can be applied to a wider range of tasks. Specifically, we show that CALI can work with a genetically inserted epitope tag; we investigate the effectiveness of alternative dyes, especially fluorescein, comparing them with the standard CALI dye, malachite green; and we study the relative efficiencies of pulsed and continuous-wave illumination. We then use fluorescein-labeled hemagglutinin antibody fragments, together with relatively low-power continuous-wave illumination to examine the effectiveness of CALI targeted to kinesin. We show that CALI can destroy kinesin activity in at least two ways: it can either result in the apparent loss of motor activity, or it can cause irreversible attachment of the kinesin enzyme to its microtubule substrate. Finally, we apply this implementation of CALI to an in vitro system of motor proteins and microtubules that is capable of self-organized aster formation. In this system, CALI can effectively perturb local structure formation by blocking or reducing the degree of aster formation in chosen regions of the sample, without influencing structure formation elsewhere.

  20. Perfluorooctanesulfonate Mediates Renal Tubular Cell Apoptosis through PPARgamma Inactivation.

    PubMed

    Wen, Li-Li; Lin, Chien-Yu; Chou, Hsiu-Chu; Chang, Chih-Cheng; Lo, Hau-Yin; Juan, Shu-Hui

    2016-01-01

    Perfluorinated chemicals (PFCs) are ubiquitously distributed in the environments including stainless pan-coating, raincoat, fire extinguisher, and semiconductor products. The PPAR family has been shown to contribute to the toxic effects of PFCs in thymus, immune and excretory systems. Herein, we demonstrated that perfluorooctanesulfonate (PFOS) caused cell apoptosis through increasing ratio of Bcl-xS/xL, cytosolic cytochrome C, and caspase 3 activation in renal tubular cells (RTCs). In addition, PFOS increased transcription of inflammatory cytokines (i.e., TNFα, ICAM1, and MCP1) by NFκB activation. Conversely, PFOS reduced the mRNA levels of antioxidative enzymes, such as glutathione peroxidase, catalase, and superoxide dismutase, as a result of reduced PPARγ transactivational activity by using reporter and chromatin immuoprecipitation (ChIP) assays. PFOS reduced the protein interaction between PPARγ and PPARγ coactivator-1 alpha (PGC1α) by PPARγ deacetylation through Sirt1 upregulation, of which the binding of PPARγ and PGC1α to a peroxisome proliferator response element (PPRE) in the promoter regions of these antioxidative enzymes was alleviated in the ChIP assay. Furthermore, Sirt1 also deacetylated p53 and then increased the binding of p53 to Bax, resulting in increased cytosolic cytochrome C. The effect of PPARγ inactivation by PFOS was validated using the PPARγ antagonist GW9662, whereas the adverse effects of PFOS were prevented by PPARγ overexpression and activators, rosiglitozone and L-carnitine, in RTCs. The in vitro finding of protective effect of L-carnitine was substantiated in vivo using Balb/c mice model subjected to PFOS challenge. Altogether, we provide in vivo and in vitro evidence for the protective mechanism of L-carnitine in eliminating PFOS-mediated renal injury, at least partially, through PPARγ activation. PMID:27171144

  1. Perfluorooctanesulfonate Mediates Renal Tubular Cell Apoptosis through PPARgamma Inactivation

    PubMed Central

    Chou, Hsiu-Chu; Chang, Chih-Cheng; Lo, Hau-Yin; Juan, Shu-Hui

    2016-01-01

    Perfluorinated chemicals (PFCs) are ubiquitously distributed in the environments including stainless pan-coating, raincoat, fire extinguisher, and semiconductor products. The PPAR family has been shown to contribute to the toxic effects of PFCs in thymus, immune and excretory systems. Herein, we demonstrated that perfluorooctanesulfonate (PFOS) caused cell apoptosis through increasing ratio of Bcl-xS/xL, cytosolic cytochrome C, and caspase 3 activation in renal tubular cells (RTCs). In addition, PFOS increased transcription of inflammatory cytokines (i.e., TNFα, ICAM1, and MCP1) by NFκB activation. Conversely, PFOS reduced the mRNA levels of antioxidative enzymes, such as glutathione peroxidase, catalase, and superoxide dismutase, as a result of reduced PPARγ transactivational activity by using reporter and chromatin immuoprecipitation (ChIP) assays. PFOS reduced the protein interaction between PPARγ and PPARγ coactivator-1 alpha (PGC1α) by PPARγ deacetylation through Sirt1 upregulation, of which the binding of PPARγ and PGC1α to a peroxisome proliferator response element (PPRE) in the promoter regions of these antioxidative enzymes was alleviated in the ChIP assay. Furthermore, Sirt1 also deacetylated p53 and then increased the binding of p53 to Bax, resulting in increased cytosolic cytochrome C. The effect of PPARγ inactivation by PFOS was validated using the PPARγ antagonist GW9662, whereas the adverse effects of PFOS were prevented by PPARγ overexpression and activators, rosiglitozone and L-carnitine, in RTCs. The in vitro finding of protective effect of L-carnitine was substantiated in vivo using Balb/c mice model subjected to PFOS challenge. Altogether, we provide in vivo and in vitro evidence for the protective mechanism of L-carnitine in eliminating PFOS-mediated renal injury, at least partially, through PPARγ activation. PMID:27171144

  2. Spontaneous reactivation of clusters of X-linked genes is associated with the plasticity of X-inactivation in mouse trophoblast stem cells.

    PubMed

    Dubois, Agnès; Deuve, Jane Lynda; Navarro, Pablo; Merzouk, Sarra; Pichard, Sylvain; Commere, Pierre-Henri; Louise, Anne; Arnaud, Danielle; Avner, Philip; Morey, Céline

    2014-02-01

    Random epigenetic silencing of the X-chromosome in somatic tissues of female mammals equalizes the dosage of X-linked genes between the sexes. Unlike this form of X-inactivation that is essentially irreversible, the imprinted inactivation of the paternal X, which characterizes mouse extra-embryonic tissues, appears highly unstable in the trophoblast giant cells of the placenta. Here, we wished to determine whether such instability is already present in placental progenitor cells prior to differentiation toward lineage-specific cell types. To this end, we analyzed the behavior of a GFP transgene on the paternal X both in vivo and in trophoblast stem (TS) cells derived from the trophectoderm of XX(GFP) blastocysts. Using single-cell studies, we show that not only the GFP transgene but also a large number of endogenous genes on the paternal X are subject to orchestrated cycles of reactivation/de novo inactivation in placental progenitor cells. This reversal of silencing is associated with local losses of histone H3 lysine 27 trimethylation extending over several adjacent genes and with the topological relocation of the hypomethylated loci outside of the nuclear compartment of the inactive X. The "reactivated" state is maintained through several cell divisions. Our study suggests that this type of "metastable epigenetic" states may underlie the plasticity of TS cells and predispose specific genes to relaxed regulation in specific subtypes of placental cells.

  3. Peptide Triazole Inactivators of HIV-1 Utilize a Conserved Two-Cavity Binding Site at the Junction of the Inner and Outer Domains of Env gp120

    PubMed Central

    Aneja, Rachna; Rashad, Adel A.; Li, Huiyuan; Sundaram, Ramalingam Venkat Kalyana; Duffy, Caitlin; Bailey, Lauren D.; Chaiken, Irwin

    2015-01-01

    We used coordinated mutagenesis, synthetic design, and flexible docking to investigate the structural mechanism of Env gp120 encounter by peptide triazole (PT) inactivators of HIV-1. Prior results demonstrated that the PT class of inhibitors suppresses binding at both CD4 and coreceptor sites on Env and triggers gp120 shedding, leading to cell-independent irreversible virus inactivation. Despite these enticing anti-HIV-1 phenotypes, structural understanding of the PT–gp120 binding mechanism has been incomplete. Here we found that PT engages two inhibitor ring moieties at the junction between the inner and outer domains of the gp120 protein. The results demonstrate how combined occupancy of two gp120 cavities can coordinately suppress both receptor and coreceptor binding and conformationally entrap the protein in a destabilized state. The two-cavity model has common features with small molecule gp120 inhibitor binding sites and provides a guide for further design of peptidomimetic HIV-1 inactivators based on the PT pharmacophore. PMID:25860784

  4. Lagrangian formulation of irreversible thermodynamics and the second law of thermodynamics

    SciTech Connect

    Glavatskiy, K. S.

    2015-05-28

    We show that the equations which describe irreversible evolution of a system can be derived from a variational principle. We suggest a Lagrangian, which depends on the properties of the normal and the so-called “mirror-image” system. The Lagrangian is symmetric in time and therefore compatible with microscopic reversibility. The evolution equations in the normal and mirror-imaged systems are decoupled and describe therefore independent irreversible evolution of each of the systems. The second law of thermodynamics follows from a symmetry of the Lagrangian. Entropy increase in the normal system is balanced by the entropy decrease in the mirror-image system, such that there exists an “integral of evolution” which is a constant. The derivation relies on the property of local equilibrium, which states that the local relations between the thermodynamic quantities in non-equilibrium are the same as in equilibrium.

  5. Irreversible electrical manipulation of magnetization on BiFeO{sub 3}-based heterostructures

    SciTech Connect

    Xu, Qingyu E-mail: jdu@nju.edu.cn; Xu, Zhenyu; He, Maocheng; Du, Jun E-mail: jdu@nju.edu.cn; Cao, Yanqiang

    2015-05-07

    We prepared several heterostructures, Co/Bi{sub 0.90}La{sub 0.10}FeO{sub 3} on surface oxidized Si or (111) SrTiO{sub 3} and NiFe/Bi{sub 0.90}La{sub 0.10}FeO{sub 3} on (001) SrTiO{sub 3} substrates using LaNiO{sub 3} as bottom electrode. With different strategies of voltage application, the exchange bias field H{sub E} decreased with increasing voltage irreversibly for all the heterostructures at room temperature. The chemical state at the NiFe/Bi{sub 0.90}La{sub 0.10}FeO{sub 3} interface was studied by X-ray photoelectron spectroscopy before and after the electrical manipulation. The oxidization of the metallic ferromagnetic layer at interface after the electrical manipulation has been confirmed, which might explain the irreversibility.

  6. Peptidyl cyclopropenones: Reversible inhibitors, irreversible inhibitors, or substrates of cysteine proteases?

    PubMed Central

    Cohen, Meital; Bretler, Uriel; Albeck, Amnon

    2013-01-01

    Peptidyl cyclopropenones were previously introduced as selective cysteine protease reversible inhibitors. In the present study we synthesized one such peptidyl cyclopropenone and investigated its interaction with papain, a prototype cysteine protease. A set of kinetics, biochemical, HPLC, MS, and 13C-NMR experiments revealed that the peptidyl cyclopropenone was an irreversible inhibitor of the enzyme, alkylating the catalytic cysteine. In parallel, this cyclopropenone also behaved as an alternative substrate of the enzyme, providing a product that was tentatively suggested to be either a spiroepoxy cyclopropanone or a gamma-lactone. Thus, a single family of compounds exhibits an unusual variety of activities, being reversible inhibitors, irreversible inhibitors and alternative substrates towards enzymes of the same family. PMID:23553793

  7. Immersion disinfection of irreversible hydrocolloid impressions with sodium hypochlorite. Part I: Microbiology.

    PubMed

    Beyerle, M P; Hensley, D M; Bradley, D V; Schwartz, R S; Hilton, T J

    1994-01-01

    Current American Dental Association infection control guidelines recommend immersion disinfection of irreversible hydrocolloid impressions, and this study further defines the parameters for use of sodium hypochlorite. Sodium hypochlorite has been shown to be an effective disinfectant for impressions; however, it has not been fully evaluated for optimum immersion time and concentration. In this study, irreversible hydrocolloid impressions contaminated with different bacteria were immersed in varying concentrations of sodium hypochlorite for 1, 5, or 10 minutes. Dilute solutions of sodium hypochlorite (0.525% or 0.0525%) produced a 4-log10 (99.99%) reduction in colony-forming units of Staphylococcus aureus, Salmonella choleraesuis, or Pseudomonas aeruginosa after 1 to 5 minutes' immersion. Full-strength sodium hypochlorite (5.25%) required 5 minutes to produce a 4-log10 reduction of Bacillus subtilis. A 4-log10 reduction of Mycobacterium bovis was not obtained under any conditions examined. PMID:7916888

  8. Immersion disinfection of irreversible hydrocolloid impressions with sodium hypochlorite. Part I: Microbiology.

    PubMed

    Beyerle, M P; Hensley, D M; Bradley, D V; Schwartz, R S; Hilton, T J

    1994-01-01

    Current American Dental Association infection control guidelines recommend immersion disinfection of irreversible hydrocolloid impressions, and this study further defines the parameters for use of sodium hypochlorite. Sodium hypochlorite has been shown to be an effective disinfectant for impressions; however, it has not been fully evaluated for optimum immersion time and concentration. In this study, irreversible hydrocolloid impressions contaminated with different bacteria were immersed in varying concentrations of sodium hypochlorite for 1, 5, or 10 minutes. Dilute solutions of sodium hypochlorite (0.525% or 0.0525%) produced a 4-log10 (99.99%) reduction in colony-forming units of Staphylococcus aureus, Salmonella choleraesuis, or Pseudomonas aeruginosa after 1 to 5 minutes' immersion. Full-strength sodium hypochlorite (5.25%) required 5 minutes to produce a 4-log10 reduction of Bacillus subtilis. A 4-log10 reduction of Mycobacterium bovis was not obtained under any conditions examined.

  9. Irreversible thermodynamic model for accelerated moment release and atmospheric radon concentration prior to large earthquakes

    NASA Astrophysics Data System (ADS)

    Kawada, Y.; Nagahama, H.; Omori, Y.; Yasuoka, Y.; Shinogi, M.

    2006-12-01

    Accelerated moment release is often preceded by large earthquakes, and defined by rate of cumulative Benioff strain following power-law time-to-failure relation. This temporal seismicity pattern is investigated in terms of irreversible thermodynamics model. The model is regulated by the Helmholtz free energy defined by the macroscopic stress-strain relation and internal state variables (generalized coordinates). Damage and damage evolution are represented by the internal state variables. In the condition, huge number of the internal state variables has each specific relaxation time, while a set of the time evolution shows a temporal power-law behavior. The irreversible thermodynamic model reduces to a fiber-bundle model and experimentally-based constitutive law of rocks, and predicts the form of accelerated moment release. Based on the model, we can also discuss the increase in atmospheric radon concentration prior to the 1995 Kobe earthquake.

  10. Irreversibility and thermoeconomics based design optimization of a ceramic heat exchanger

    SciTech Connect

    Ranasinghe, J.; Aceves-Saborio, S.; Reistad, G.M. )

    1989-10-01

    This paper illustrates the optimization procedure for heat exchangers residing in complex power plants. A specific case of optimizing a new technology ceramic heat exchanger, which is part of the complex power plant, is shown. The heat exchanger design methods presented are based on two different objective functions, namely, a modified irreversibility rate based objective function proposed by the authors in earlier work and an objective function based on thermoeconomics. This paper also extends existing work by illustrating a method to obtain the cost coefficients for thermoeconomic optimization, based on the use of an overall plant simulation model. A discussion on possible methods of improving the design guideposts obtained from irreversibility minimization analysis is presented.

  11. Inactivation of Gram-Negative Bacteria by Lysozyme, Denatured Lysozyme, and Lysozyme-Derived Peptides under High Hydrostatic Pressure

    PubMed Central

    Masschalck, Barbara; Van Houdt, Rob; Van Haver, Ellen G. R.; Michiels, Chris W.

    2001-01-01

    We have studied the inactivation of six gram-negative bacteria (Escherichia coli, Pseudomonas fluorescens, Salmonella enterica serovar Typhimurium, Salmonella enteritidis, Shigella sonnei, and Shigella flexneri) by high hydrostatic pressure treatment in the presence of hen egg-white lysozyme, partially or completely denatured lysozyme, or a synthetic cationic peptide derived from either hen egg white or coliphage T4 lysozyme. None of these compounds had a bactericidal or bacteriostatic effect on any of the tested bacteria at atmospheric pressure. Under high pressure, all bacteria except both Salmonella species showed higher inactivation in the presence of 100 μg of lysozyme/ml than without this additive, indicating that pressure sensitized the bacteria to lysozyme. This extra inactivation by lysozyme was accompanied by the formation of spheroplasts. Complete knockout of the muramidase enzymatic activity of lysozyme by heat treatment fully eliminated its bactericidal effect under pressure, but partially denatured lysozyme was still active against some bacteria. Contrary to some recent reports, these results indicate that enzymatic activity is indispensable for the antimicrobial activity of lysozyme. However, partial heat denaturation extended the activity spectrum of lysozyme under pressure to serovar Typhimurium, suggesting enhanced uptake of partially denatured lysozyme through the serovar Typhimurium outer membrane. All test bacteria were sensitized by high pressure to a peptide corresponding to amino acid residues 96 to 116 of hen egg white, and all except E. coli and P. fluorescens were sensitized by high pressure to a peptide corresponding to amino acid residues 143 to 155 of T4 lysozyme. Since they are not enzymatically active, these peptides probably have a different mechanism of action than all lysozyme polypeptides. PMID:11133464

  12. Irreversible visual loss and optic nerve dysfunction associated with central retinal vein occlusion in Waldenstrom Macroglobulinemia.

    PubMed

    Fadilah, S A W; Muhaya, M; Azlin, I

    2007-10-01

    Irreversible optic nerve dysfunction associated with central retinal vein occlusion (CRVO) is an unusual but important complication of Waldenstrom Macroglobulinemia (WM). Acute visual loss in CRVO is mainly due the severe macular oedema. However, ischaemic optic neuropathy needs to be considered in patients with CRVO when, (i) there is a relative afferent papillary defect and central scotoma, (ii) the visual acuity is not consistent with the retinal pathology, and (iii) the visual defects persisted despite resolution of macular oedema following treatment of the hyperviscosity state. The ischaemic type of CRVO is associated with a poor visual prognosis and the presenting visual acuity has a prognostic role. We report the first description of irreversible unilateral optic nerve damage associated with CRVO in a patient with WM. PMID:18551947

  13. On heat equation in the framework of classic irreversible thermodynamics with internal variables

    NASA Astrophysics Data System (ADS)

    Ciancio, Vincenzo; Restuccia, Liliana

    2016-06-01

    In this paper, we show that, using a procedure of classical irreversible thermodynamics (CIT) with internal variables, it is possible to describe the relaxation of thermal phenomena, obtaining some well known results of extended irreversible thermodynamics (EIT). In particular, we introduce as internal variables a vector and a second rank tensor, that influence the thermal transport phenomena, and we derive in the anisotropic and isotropic case, the phenomenological equations for these variables. In the case, in which the medium is isotropic, it is obtained that the total heat flux can be split in two parts: a first contribution J(0), governed by Fourier law, and a second contribution J(1), obeying Maxwell-Cattaneo-Vernotte (MCV) equation, in which a relaxation time is present. The obtained results may have applications in describing the thermal behavior in nanosystems (semiconductors, nanotubes,…), where the phenomena are fast and there are high-frequency thermal waves.

  14. Immersion disinfection of irreversible hydrocolloid impressions with sodium hypochlorite. Part II: Effect on gypsum.

    PubMed

    Vandewalle, K S; Charlton, D G; Schwartz, R S; Reagan, S E; Koeppen, R G

    1994-01-01

    This study investigated the effects of various immersion times and concentrations of sodium hypochlorite on irreversible hydrocolloid impressions and resultant gypsum casts. Irreversible hydrocolloid impressions of a test die were immersed for 1, 5, or 10 minutes in water (control), 5.25%, 0.525%, and 0.0525% sodium hypochlorite and then cast in a Type III stone and a Type V stone. Each stone specimen was evaluated for detail reproduction, dimensional change, surface roughness, and surface hardness. The results indicated that impressions may be immersed in sodium hypochlorite for any of the experimental times and concentrations without negative effects on Type V stone casts. However, immersion of impressions in 5.25% sodium hypochlorite causes some surface deterioration on Type III stone casts. PMID:7993542

  15. Suppression of irreversible capacity loss in Li-rich layered oxide by fluorine doping

    NASA Astrophysics Data System (ADS)

    Song, Jay Hyok; Kapylou, Andrei; Choi, Hee Sung; Yu, Byong Yong; Matulevich, Evegeniya; Kang, Sun Ho

    2016-05-01

    Li[Li1/6Ni1/6Co1/6Mn1/2]O2-xFx (x = 0.00 to 0.07) materials were synthesized with low temperature heat treatment (700 °C) and their electrochemical performances were evaluated. With the addition of fluorine, the reversible capacity significantly increased as the irreversibility was suppressed during the first cycle. The reduction of irreversibility was mainly attributed to the enhanced first cycle efficiency of Li2MnO3-like component after the fluorine addition. By combining results of the X-ray diffraction (XRD), secondary ion mass spectrometry (SIMS), In-situ X-ray absorption spectroscopy (XAS) analyses, and first principle calculations, it was proposed that the presence of fluorine facilitated the reduction of cobalt and manganese ions in Li-rich layered oxide, and that the reduced transition metal (TM) ions suppressed structural changes.

  16. In vivo efficacy of natural product-inspired irreversible kinase inhibitors.

    PubMed

    Barluenga, Sofia; Jogireddy, Rajamalleswaramma; Koripelly, Girish K; Winssinger, Nicolas

    2010-08-16

    Hypothemycin and related resorcylic acid lactones (RAL) bearing a cis-enone moiety have emerged as an alternative pharmacophore to heterocyclic motifs for kinase inhibition, and are endowed with a unique selectivity filter based on the irreversible reaction with a subset of the kinome bearing a suitably positioned cysteine residue. Two prototypical examples of "edited" RAL were evaluated for antitumoral, antimetastatic and antiangiogenic efficacy in an orthotopic murine renal cell carcinoma (RENCA) model. Both compounds (3 and 5) are good inhibitors of VEGFRs in vitro, and inhibited tumor growth in vivo with comparable efficacy to sunitinib, an FDA-approved VEGFRs inhibitor. Compound 3 promoted lung metastasis to a similar extent as sunitinib, while compound 5 strongly inhibited lung metastasis. This study attests to the potential of irreversible kinase inhibitors and molecular editing of natural pharmacophores and provides encouraging results to a clinically significant problem. PMID:20623569

  17. Work statistics, irreversible heat and correlations build-up in joining two spin chains

    NASA Astrophysics Data System (ADS)

    Apollaro, T. J. G.; Francica, Gianluca; Paternostro, Mauro; Campisi, Michele

    2015-10-01

    We investigate the influences of quantum many-body effects, such as criticality and the existence of factorization fields, in the thermodynamic cost of establishing a bonding link between two independent quantum spin chains. We provide a physical interpretation of the behavior of irreversible work spent in such a process by linking the phenomenology of such quantities to the properties of the spectrum of the system.

  18. Irreversible decay of nonlocal entanglement via a reservoir of a single degree of freedom

    SciTech Connect

    Vaglica, A.; Vetri, G.

    2007-06-15

    Recently, it has been realized that nonlocal disentanglement may take a finite time as opposite to the asymptotic decay of local coherences. We find in this paper that a sudden irreversible death of entanglement takes place in a two atom optical Stern-Gerlach model. In particular, the one degree nondissipative environment here considered suddenly destroys the initial entanglement of any Bell's states vertical bar {phi}{sup {+-}}> superposition.

  19. Rapid irreversible encephalopathy associated with anti-D immune globulin treatment for idiopathic thrombocytopenic purpura.

    PubMed

    Christopher, Kenneth; Horkan, Clare; Barb, Ilie T; Arbelaez, Christian; Hodgdon, Travis A; Yodice, Paul C

    2004-11-01

    Intravenous Rho (D) immune globulin (IV RhIG, WinRho SDF) has been shown to be a safe treatment for idiopathic thrombocytopenic purpura (ITP). Common side effects of IV RhIG include mild hemolysis, febrile reaction, and headache. Significant hemolysis with renal impairment following IV RhIG has been reported. We report a case of irreversible encephalopathy 48 hr following an infusion of IV RhIG for treatment of ITP.

  20. Microbiome of Deep Dentinal Caries Lesions in Teeth with Symptomatic Irreversible Pulpitis

    PubMed Central

    Rôças, Isabela N.; Rachid, Caio T. C. C.; Lima, Kenio C.; Assunção, Isauremi V.; Gomes, Patrícia N.; Siqueira, José F.

    2016-01-01

    This study used a next-generation sequencing approach to identify the bacterial taxa occurring in the advanced front of caries biofilms associated with pulp exposure and irreversible pulpitis. Samples were taken from the deepest layer of dentinal caries lesions associated with pulp exposure in 10 teeth diagnosed with symptomatic irreversible pulpitis. DNA was extracted and the microbiome was characterized on the basis of the V4 hypervariable region of the 16S rRNA gene by using paired-end sequencing on Illumina MiSeq device. Bacterial taxa were mapped to 14 phyla and 101 genera composed by 706 different OTUs. Three phyla accounted for approximately 98% of the sequences: Firmicutes, Actinobacteria and Proteobacteria. These phyla were also the ones with most representatives at the species level. Firmicutes was the most abundant phylum in 9/10 samples. As for genera, Lactobacillus accounted for 42.3% of the sequences, followed by Olsenella (13.7%), Pseudoramibacter (10.7%) and Streptococcus (5.5%). Half of the samples were heavily dominated by Lactobacillus, while in the other half lactobacilli were in very low abundance and the most dominant genera were Pseudoramibacter, Olsenella, Streptococcus, and Stenotrophomonas. High bacterial diversity occurred in deep dentinal caries lesions associated with symptomatic irreversible pulpitis. The microbiome could be classified according to the relative abundance of Lactobacillus. Except for Lactobacillus species, most of the highly prevalent and abundant bacterial taxa identified in this study have been commonly detected in infected root canals. The detected taxa can be regarded as candidate pathogens for irreversible pulpitis and possibly the pioneers in pulp invasion to initiate endodontic infection. PMID:27135405