Sample records for affecting early enzymatic

  1. Enzymatic biodiesel synthesis. Key factors affecting efficiency of the process

    Energy Technology Data Exchange (ETDEWEB)

    Szczesna Antczak, Miroslawa; Kubiak, Aneta; Antczak, Tadeusz; Bielecki, Stanislaw [Institute of Technical Biochemistry, Faculty of Biotechnology and Food Sciences, Technical University of Lodz, Stefanowskiego 4/10, 90-924 Lodz (Poland)


    Chemical processes of biodiesel production are energy-consuming and generate undesirable by-products such as soaps and polymeric pigments that retard separation of pure methyl or ethyl esters of fatty acids from glycerol and di- and monoacylglycerols. Enzymatic, lipase-catalyzed biodiesel synthesis has no such drawbacks. Comprehension of the latter process and an appreciable progress in production of robust preparations of lipases may soon result in the replacement of chemical catalysts with enzymes in biodiesel synthesis. Engineering of enzymatic biodiesel synthesis processes requires optimization of such factors as: molar ratio of substrates (triacylglycerols: alcohol), temperature, type of organic solvent (if any) and water activity. All of them are correlated with properties of lipase preparation. This paper reports on the interplay between the crucial parameters of the lipase-catalyzed reactions carried out in non-aqueous systems and the yield of biodiesel synthesis. (author)

  2. Enzymatic Antioxidant Systems in Early Anaerobes: Theoretical Considerations (United States)

    Ślesak, Ireneusz; Ślesak, Halina; Zimak-Piekarczyk, Paulina; Rozpądek, Piotr


    It is widely accepted that cyanobacteria-dependent oxygen that was released into Earth's atmosphere ca. 2.5 billion years ago sparked the evolution of the aerobic metabolism and the antioxidant system. In modern aerobes, enzymes such as superoxide dismutases (SODs), peroxiredoxins (PXs), and catalases (CATs) constitute the core of the enzymatic antioxidant system (EAS) directed against reactive oxygen species (ROS). In many anaerobic prokaryotes, the superoxide reductases (SORs) have been identified as the main force in counteracting ROS toxicity. We found that 93% of the analyzed strict anaerobes possess at least one antioxidant enzyme, and 50% have a functional EAS, that is, consisting of at least two antioxidant enzymes: one for superoxide anion radical detoxification and another for hydrogen peroxide decomposition. The results presented here suggest that the last universal common ancestor (LUCA) was not a strict anaerobe. O2 could have been available for the first microorganisms before oxygenic photosynthesis evolved, however, from the intrinsic activity of EAS, not solely from abiotic sources.

  3. Applications of a single-molecule detection in early disease diagnosis and enzymatic reaction study

    Energy Technology Data Exchange (ETDEWEB)

    Li, Jiangwei [Iowa State Univ., Ames, IA (United States)


    polyclonal detector antibody. The capture antibody was covalently immobilized on modified glass slides. The detector antibody was conjugated with AF532 labeled secondary antibody prior to being used as probe for the antigen. Imaging was performed with a TIRF system. This technique is demonstrated for detecting HIV-1 p24 antigen down to 0.1 pg/mL with a dynamic range of over 4 orders of magnitude. A Langmuir isotherm fit the molecule count dependence on the target concentration. The results also showed that neither sensitivity nor dynamic range was affected by the biological matrix. SMISA is therefore a promising approach for the early diagnosis of virus-induced diseases. Single-molecule enzymatic kinetics and enantioselectivity were monitored in real time by using TIRF microscopy. AF532 labeled poly-L-lysine (PLL) or poly-D-lysine (PDL) was covalently immobilized on a dithiobis (succinimidyl undecanoate) self-assembled monolayer (DSU SAM). Chain shortening due to enzymatic hydrolysis resulted in the reduction of the individual fluorescence intensities. A broad distribution was obtained when 100 single-molecule half-lives were analyzed. However, the detailed hydrolysis process involved also a long-lived component and an induction period that varied significantly among molecules. Charge and steric heterogeneity at the surface are responsible for these features.

  4. Particle size distribution of rice flour affecting the starch enzymatic hydrolysis and hydration properties. (United States)

    de la Hera, Esther; Gomez, Manuel; Rosell, Cristina M


    Rice flour is becoming very attractive as raw material, but there is lack of information about the influence of particle size on its functional properties and starch digestibility. This study evaluates the degree of dependence of the rice flour functional properties, mainly derived from starch behavior, with the particle size distribution. Hydration properties of flours and gels and starch enzymatic hydrolysis of individual fractions were assessed. Particle size heterogeneity on rice flour significantly affected functional properties and starch features, at room temperature and also after gelatinization; and the extent of that effect was grain type dependent. Particle size heterogeneity on rice flour induces different pattern in starch enzymatic hydrolysis, with the long grain having slower hydrolysis as indicated the rate constant (k). No correlation between starch digestibility and hydration properties or the protein content was observed. It seems that in intact granules interactions with other grain components must be taken into account. Overall, particle size fractionation of rice flour might be advisable for selecting specific physico-chemical properties.

  5. Early growth trajectories affect sexual responsiveness. (United States)

    Lee, Who-Seung; Metcalfe, Neil B; Réale, Denis; Peres-Neto, Pedro R


    The trajectory of an animal's growth in early development has been shown to have long-term effects on a range of life-history traits. Although it is known that individual differences in behaviour may also be related to certain life-history traits, the linkage between early growth or development and individual variation in behaviour has received little attention. We used brief temperature manipulations, independent of food availability, to stimulate compensatory growth in juvenile three-spined sticklebacks Gasterosteus aculeatus. Here, we examine how these manipulated growth trajectories affected the sexual responsiveness of the male fish at the time of sexual maturation, explore associations between reproductive behaviour and investment and lifespan and test whether the perceived time stress (until the onset of the breeding season) influenced such trade-offs. We found a negative impact of growth rate on sexual responsiveness: fish induced (by temperature manipulation) to grow slowest prior to the breeding season were consistently quickest to respond to the presence of a gravid female. This speed of sexual responsiveness was also positively correlated with the rate of development of sexual ornaments and time taken to build a nest. However, after controlling for effects of growth rate, those males that had the greatest sexual responsiveness to females had the shortest lifespan. Moreover, the time available to compensate in size before the onset of the breeding season (time stress) affected the magnitude of these effects. Our results demonstrate that developmental perturbations in early life can influence mating behaviour, with long-term effects on longevity.

  6. Structural features affecting the enzymatic digestibility of pine wood pretreated with ionic liquids. (United States)

    Torr, Kirk M; Love, Karen T; Simmons, Blake A; Hill, Stefan J


    Pretreating lignocellulosic biomass with certain ionic liquids results in structural and chemical changes that make the biomass more digestible by enzymes. In this study, pine wood was pretreated with 1-ethyl-3-methylimidazolium chloride/acetate ([C2 mim]Cl and [C2 mim][OAc]) at different temperatures to investigate the relative importance of substrate features, such as accessible surface area, cellulose crystallinity, and lignin content, on enzymatic digestibility. The ionic liquid pretreatments resulted in glucan conversions ranging from 23% to 84% on saccharification of the substrates, with [C2 mim][OAc] being more effective than [C2 mim]Cl. The pretreatments resulted in no delignification of the wood, some loss of cellulose crystallinity under certain conditions, and varying levels of increased surface area. Enzymatic digestibility closely correlated with accessible surface area and porosity measurements obtained using Simons' staining and thermoporosimetry techniques. Increased accessible surface area was identified as the principal structural feature responsible for the improved enzymatic digestibility.

  7. Time of harvest affects the yield of soluble polysaccharides extracted enzymatically from potato pulp

    DEFF Research Database (Denmark)

    Ravn, Helle Christine; Sørensen, Ole Bandsholm; Meyer, Anne S.


    potato polysaccharides applying a recently developed enzymatic process using 1.0% (w/w) [enzyme/substrate (E/S)] pectin lyase from Aspergillus nidulans and 1.0% (w/w) [E/S] polygalacturonase from A. aculeatus at 60 °C, 100 mM citric acid, pH 6.0 for 1 min. Seven samples drawn within the potato starch...

  8. Enzymatic debridement for the treatment of severely burned upper extremities – early single center experiences


    Cordts, Tomke; Horter, Johannes; Vogelpohl, Julian; Kremer, Thomas; Kneser, Ulrich; Hernekamp, Jochen-Frederick


    Background Severe burns of hands and arms are complex and challenging injuries. The Standard of care (SOC) – necrosectomy with skin grafting – is often associated with poor functional or aesthetic outcome. Enzymatic debridement (ED) is considered one promising alternative but, until recently, results proved to be highly variable. Methods Between 04/2014 and 04/2015, 16 patients with deep partial- to full-thickness burns of the upper extremities underwent enzymatic debridement (ED) in our Burn...

  9. Lignocellulose pretreatment technologies affect the level of enzymatic cellulose oxidation by LPMO

    DEFF Research Database (Denmark)

    Rodríguez-Zúñiga, Ursula Fabiola; Cannella, David; de Campos Giordano, Roberto


    Sugarcane bagasse, corn stover, and wheat straw are among the most available resources for production of cellulosic ethanol. For these biomasses we study the influence of pre-treatment methods on the chemical composition, as well as on the subsequent reactions of enzymatic hydrolysis and oxidation...... of cellulose. The applied pre-treatment methods are organosolv, hydrothermal, and alkaline. Hydrothermally pretreated wheat straw gave the highest cellulose conversion with 80% glucose yield and 0.8% oxidized cellulose products. Recent studies have shown that lignin is able to boost the activity...

  10. Dynamics of learner affective development in early FLL

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    Jelena Mihaljević Djigunović


    Full Text Available Affective learner factors were first considered as a cause of success in language learning. This was followed by a change in approach and recently authors (e.g., Edelenbos, Johnstone, & Kubanek, 2006 have considered them an important outcome, especially in early foreign language learning (FLL. Current research into affective learner factors in early FLL tries to catch the developmental aspects too, and studies are emerging that take a contextual view as well. This paper describes a study on affective characteristics of young FL learners that combines the developmental and contextual perspectives. Using the case study methodology the author analyses the affective profiles of three young learners of English as a foreign language who were followed for 4 years. The analyses are done taking into account their immediate language learning environment, home support, out-of-school exposure to English and language achievement. The findings suggest that affective learner factors contribute to the dynamic complexity of early FLL.

  11. How metal substitution affects the enzymatic activity of catechol-o-methyltransferase.

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    Manuel Sparta

    Full Text Available Catechol-O-methyltransferase (COMT degrades catecholamines, such as dopamine and epinephrine, by methylating them in the presence of a divalent metal cation (usually Mg(II, and S-adenosyl-L-methionine. The enzymatic activity of COMT is known to be vitally dependent on the nature of the bound metal: replacement of Mg(II with Ca(II leads to a complete deactivation of COMT; Fe(II is slightly less than potent Mg(II, and Fe(III is again an inhibitor. Considering the fairly modest role that the metal plays in the catalyzed reaction, this dependence is puzzling, and to date remains an enigma. Using a quantum mechanical / molecular mechanical dynamics method for extensive sampling of protein structure, and first principle quantum mechanical calculations for the subsequent mechanistic study, we explicate the effect of metal substitution on the rate determining step in the catalytic cycle of COMT, the methyl transfer. In full accord with experimental data, Mg(II bound to COMT is the most potent of the studied cations and it is closely followed by Fe(II, whereas Fe(III is unable to promote catalysis. In the case of Ca(II, a repacking of the protein binding site is observed, leading to a significant increase in the activation barrier and higher energy of reaction. Importantly, the origin of the effect of metal substitution is different for different metals: for Fe(III it is the electronic effect, whereas in the case of Ca(II it is instead the effect of suboptimal protein structure.

  12. Alkaline-assisted screw press pretreatment affecting enzymatic hydrolysis of wheat straw. (United States)

    Yan, Qingqi; Wang, Yumei; Rodiahwati, Wawat; Spiess, Antje; Modigell, Michael


    Screw press processing of biomass can be considered as a suitable mechanically based pretreatment for biofuel production since it disrupts the structure of lignocellulosic biomass with high shear and pressure forces. The combination with chemical treatment has been suggested to increase the conversion of lignocellulosic biomass to fermentable sugars. Within the study, the synergetic effect of alkaline (sodium hydroxide, NaOH) soaking and screw press pretreatment on wheat straw was evaluated based on, e.g., sugar recovery and energy efficiency. After alkaline soaking (at 0.1 M for 30 min) and sequential screw press pretreatment with various screw press configurations and modified screw barrel, the lignin content of pretreated wheat straw was quantified. In addition, the structure of pretreated wheat straw was investigated by scanning electron microscopy and measurement of specific surface area. It could be shown that removal of lignin is more important than increase of surface area of the biomass to reach a high sugar recovery. The rate constant of the enzymatic hydrolysis increased from 1.1 × 10(-3) 1/h for the non-treated material over 2.3 × 10(-3) 1/h for the alkaline-soaked material to 26.9 × 10(-3) 1/h for alkaline-assisted screw press pretreated material, indicating a nearly 25-fold improvement of the digestibility by the combined chemo-mechanical pretreatment. Finally, the screw configuration was found to be an important factor for improving the sugar recovery and for reducing the specific energy consumption of the screw press pretreatment.

  13. Traits affecting early season nitrogen uptake in nine legume species

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    Elana Dayoub


    Full Text Available Legume crops are known to have low soil N uptake early in their life cycle, which can weaken their ability to compete with other species, such as weeds or other crops in intercropping systems. However, there is limited knowledge on the main traits involved in soil N uptake during early growth and for a range of species. The objective of this research was to identify the main traits explaining the variability among legume species in soil N uptake and to study the effect of the soil mineral N supply on the legume strategy for the use of available N sources during early growth. Nine legume species were grown in rhizotrons with or without N supply. Root expansion, shoot and root biomass, nodule establishment, N2 fixation and mineral soil N uptake were measured. A large interspecific variability was observed for all traits affecting soil N uptake. Root lateral expansion and early biomass in relation to seed mass were the major traits influencing soil N uptake regardless of the level of soil N availability. Fenugreek, lentil, alfalfa, and common vetch could be considered weak competitors for soil N due to their low plant biomass and low lateral root expansion. Conversely, peanut, pea, chickpea and soybean had a greater soil N uptake. Faba bean was separated from other species having a higher nodule biomass, a higher N2 fixation and a lower seed reserve depletion. Faba bean was able to simultaneously fix N2 and take up soil N. This work has identified traits of seed mass, shoot and root biomass, root lateral expansion, N2 fixation and seed reserve depletion that allowing classification of legume species regarding their soil N uptake ability during early growth.

  14. Factors affecting daughter cells' arrangement during the early bacterial divisions.

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    Pin-Tzu Su

    Full Text Available On agar plates, daughter cells of Escherichia coli mutually slide and align side-by-side in parallel during the first round of binary fission. This phenomenon has been previously attributed to an elastic material that restricts apparently separated bacteria from being in string. We hypothesize that the interaction between bacteria and the underneath substratum may affect the arrangement of the daughter bacteria. To test this hypothesis, bacterial division on hyaluronic acid (HA gel, as an alternative substratum, was examined. Consistent with our proposition, the HA gel differs from agar by suppressing the typical side-by-side alignments to a rare population. Examination of bacterial surface molecules that may contribute to the daughter cells' arrangement yielded an observation that, with disrupted lpp, the E. coli daughter cells increasingly formed non-typical patterns, i.e. neither sliding side-by-side in parallel nor forming elongated strings. Therefore, our results suggest strongly that the early cell patterning is affected by multiple interaction factors. With oscillatory optical tweezers, we further demonstrated that the interaction force decreased in bacteria without Lpp, a result substantiating our notion that the side-by-side sliding phenomenon directly reflects the strength of in-situ interaction between bacteria and substratum.

  15. Enzymatic characterization of germination-specific cysteine protease-1 expressed transiently in cotyledons during the early phase of germination. (United States)

    Tsuji, Akihiko; Tsukamoto, Kana; Iwamoto, Keiko; Ito, Yuka; Yuasa, Keizo


    Papain-like cysteine protease activity that shows a unique transient expression profile in cotyledons of daikon radish during germination was detected. The enzyme showed a distinct elution pattern on DEAE-cellulose compared with cathepsin B-like and Responsive to dessication-21 cysteine protease. Although this activity was not detected in seed prior to imbibition, the activity increased markedly and reached a maximum at 2 days after imbibition and then decreased rapidly and completely disappeared after 5 days. Using cystatin-Sepharose, the 26 kDa cysteine protease (DRCP26) was isolated from cotyledons at 2 days after imbibition. The deduced amino acid sequence from the cDNA nucleotide sequence indicated that DRCP26 is an orthologue of Arabidopsis unidentified protein, germination-specific cysteine protease-1, belonging to the C1 family of cysteine protease predicted from genetic information. In an effort to characterize the enzymatic properties of DRCP26, the enzyme was purified to homogeneity from cotyledons at 48 h after imbibition. The best synthetic substrate for the enzyme was carbobenzoxy-Phe-Arg-4-methylcoumaryl-7-amide. All model peptides were digested to small peptides by the enzyme, suggesting that DRCP26 possesses broad cleavage specificity. These results indicated that DRCP26 plays a role in the mobilization of storage proteins in the early phase of seed germination.

  16. Gestational length affects neurocognition in early-onset schizophrenia. (United States)

    Teigset, Charlotte M; Mohn, Christine; Rund, Bjørn Rishovd


    Obstetric complications (OC) have been linked to an increased risk for schizophrenia in offspring, especially in early-onset schizophrenia (EOS). Extensive cognitive deficits occur in EOS, although no study has yet to investigate the relationship between OC and cognition in EOS. This study aims to examine the frequency of OC in EOS compared to controls, and also investigates the relationship between OC and neurocognitive dysfunction in the two groups. Nineteen EOS patients and 53 healthy controls were tested with the MATRICS Consensus Cognitive Battery (MCCB), and the cognitive measures were combined with OC data from the Norwegian Birth Registry. The results indicated no group differences in OC in EOS and healthy controls, but a shorter gestational length in the EOS group led to significant decreases in the overall neurocognitive composite score, and in processing speed. This suggests that the poorer neuropsychological performances commonly found in EOS may be partly attributable to the length of gestation. The worsened neurocognitive functioning did not appear among controls, so gestational length had a different impact on the two groups. Our findings indicated that a shorter gestational length did not increase the risk for developing EOS, but did significantly affect the cognitive difficulties in this group.

  17. Early parental death and risk of hospitalization for affective disorder in adulthood

    DEFF Research Database (Denmark)

    Appel, Charlotte Welling; Johansen, Christoffer; Deltour, Isabelle


    Early parental death is one of the most stressful childhood life events and may influence subsequent psychological health. We investigated the association between early parental loss and risk of hospitalization for an affective disorder in adulthood.......Early parental death is one of the most stressful childhood life events and may influence subsequent psychological health. We investigated the association between early parental loss and risk of hospitalization for an affective disorder in adulthood....

  18. Biomass enzymatic saccharification is determined by the non-KOH-extractable wall polymer features that predominately affect cellulose crystallinity in corn. (United States)

    Jia, Jun; Yu, Bin; Wu, Leiming; Wang, Hongwu; Wu, Zhiliang; Li, Ming; Huang, Pengyan; Feng, Shengqiu; Chen, Peng; Zheng, Yonglian; Peng, Liangcai


    Corn is a major food crop with enormous biomass residues for biofuel production. Due to cell wall recalcitrance, it becomes essential to identify the key factors of lignocellulose on biomass saccharification. In this study, we examined total 40 corn accessions that displayed a diverse cell wall composition. Correlation analysis showed that cellulose and lignin levels negatively affected biomass digestibility after NaOH pretreatments at pbiomass saccharification after pretreatments with NaOH and H2SO4 at three concentrations. Notably, despite that the non-KOH-extractable residues covered 12%-23% hemicelluloses and lignin of total biomass, their wall polymer features exhibited the predominant effects on biomass enzymatic hydrolysis including Ara substitution degree of xylan (reverse Xyl/Ara) and S/G ratio of lignin. Furthermore, the non-KOH-extractable polymer features could significantly affect lignocellulose crystallinity at pbiomass digestibility. Hence, this study could suggest an optimal approach for genetic modification of plant cell walls in bioenergy corn.

  19. Older Siblings Affect Gut Microbiota Development in Early Childhood

    DEFF Research Database (Denmark)

    Laursen, Martin Frederik; Zachariassen, Gitte; Bahl, Martin Iain

    .006) at 18 months. Further, having older siblings was associated with increased relative abundance of several bacterial taxa at both 9 and 18 months of age. Compared to the effect of having siblings, presence of household furred pets and early life infections had less pronounced effects on the gut microbiota....... Gut microbiota characteristics were not significantly associated with cumulative occurrence of eczema and asthmatic bronchitis during the first three years of life. Conclusions: Presence of older siblings is associated with increased gut microbial diversity and richness during early childhood, which...... could contribute to the substantiation of the hygiene hypothesis. However, no associations were found between gut microbiota and atopic symptoms of eczema and asthmatic bronchitis during early childhood and thus further studies are required to elucidate whether sibling-associated gut microbial changes...

  20. Steam explosion of Brewer's spent grain improves enzymatic digestibility of carbohydrates and affects solubility and stability of proteins. (United States)

    Kemppainen, K; Rommi, K; Holopainen, U; Kruus, K


    Steam explosion was studied as a means to improve the enzymatic digestibility of carbohydrates in Brewer's spent grain, a protein and lipid-rich lignocellulosic by-product of the brewing industry. Having temperature, treatment time and the presence of acid catalyst as variables, a treatment at 200 °C for 10 min without an acid catalyst was found to be the most efficient, dissolving 12.1 % of the dry matter. Mainly oligomeric non-cellulosic glucan and arabinoxylan were dissolved, and the remaining insoluble carbohydrates could be efficiently hydrolysed by an enzyme cocktail (75 % hydrolysis yield). The process also caused partial protein degradation and dissolved over a third of the total nitrogen. Meanwhile, the insoluble protein appeared to become more strongly associated with acid-insoluble lignin. Compositional changes observed in the proteins and carbohydrates were supported by the results of epifluorescence microscopy. The process yielded three chemically different fractions which could serve as biorefinery products or intermediates.

  1. Early embryogenesis in zebrafish is affected by bisphenol A exposure

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    William K. F. Tse


    Exposure of a developing embryo or fetus to endocrine disrupting chemicals (EDCs has been hypothesized to increase the propensity of an individual to develop a disease or dysfunction in his/her later life. Although it is important to understand the effects of EDCs on early development in animals, sufficient information about these effects is not available thus far. This is probably because of the technical difficulties in tracing the continuous developmental changes at different stages of mammalian embryos. The zebrafish, an excellent model currently used in developmental biology, provides new insights to the field of toxicological studies. We used the standard whole-mount in situ hybridization screening protocol to determine the early developmental defects in zebrafish embryos exposed to the ubiquitous pollutant, bisphenol A (BPA. Three stages (60–75% epiboly, 8–10 somite, and prim-5 were selected for in situ screening of different molecular markers, whereas BPA exposure altered early dorsoventral (DV patterning, segmentation, and brain development in zebrafish embryos within 24 hours of exposure.

  2. How can Early Life Flavor Experiences Affect Food Preferences?

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    Seray Kabaran


    Full Text Available Early experiences with food flavors lead up to children’s food preferences and develop long-lasting flavor preferences and healthy eating habits. The process of flavor learning begins during pregnancy by the foods eaten by mothers which pass to the amniotic fluid. Some flavors coming from the mother’s diet that pass to the amniotic fluid are detected by the fetus. This flavor learning continues after birth with breastfeeding, since the human milk is composed of flavors which reflect the foods consumed by the mother. Flavor composition in human milk which changes during the course of lactation due to the transmission of flavors into human milk enables the baby to get used to new flavors. Therefore, breastfed infants are likely to accept flavor changes and novel flavors more than formula fed infants. Flavor learning continues with starting complementary foods and eating the foods directly. Exposure to a specific flavor and repeated exposures in different times may facilitate the acceptance of this flavor. Furthermore, infants exposed to a variety of foods accept new flavors than infants following a monotonous diet. This underlines the importance of promoting the access to a variety of foods in early childhood. In conclusion, pregnant and lactating women should have variety of healthy foods in their diets with a variety of flavors and they should be encouraged to breastfeed their babies. In addition, starting complementary foods and early repeated exposure to a wide variety of healthy foods, the infants should get familiar with the flavor of healthy and various food. These factors may influence the development of healthy food preferences and healthy eating habits in later periods.

  3. Clinical factors adversely affecting early outcome after brain infarction

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    Charles André


    Full Text Available PURPOSE AND METHODS: One-hundred-and-nine consecutive patients admitted during the acute phase of a CT-confirmed brain infarction (BI were studied. Putative adverse influence of demographic and stroke risk factors, previous medical history, clinical presentation, initial and follow-up neurological examination, initial general evaluation, laboratory findings, chest X-ray and electrocardiographic findings, treatment, and topography and etiology of the ischemic insult was analysed. The end-point for assessment was early death (within 30 days. Statistical analysis was performed with univariate analysis and multiple regression. RESULTS: The main adverse factors related to an increased death risk during the first 30 days were, in decreasing order of importance: coma 48-72 hours after admission; stroke occuring in already hospitalized patients; Babinski sign on admission; minor degrees of impairment of consciousness 48-72 hours after admission; stroke related to large artery atherothrombosis and to embolism; a history of early impairment of consciousness; cardiac failure on admission. In 53 lucid patients on admission, only a history of congestive heart failure (CHF was associated with a reduced survival rate. In 56 patients with impaired consciousness, the presence of a Babinski sign increased death risk, but the main factor predicting a high case-fatality rate was the persistence of consciousness disturbances after48-72 hours. CONCLUSIONS: The presence of impairment of consciousness, especially coma, 2-3 days after disease onset, and a history of CHF greatly increase the early case fatality rate in patients with acute BI presenting with or without consciousness disturbances at admission, respectively. The use of a prognostic algorythm considering these few variables seems to predict the approximate 30-day fatality rates.

  4. Maillard reaction and enzymatic browning affect the allergenicity of Pru av 1, the major allergen from cherry (Prunus avium). (United States)

    Gruber, Patrick; Vieths, Stefan; Wangorsch, Andrea; Nerkamp, Jörg; Hofmann, Thomas


    The influence of thermal processing and nonenymatic as well as polyphenoloxidase-catalyzed browning reaction on the allergenicity of the major cherry allergen Pru av 1 was investigated. After thermal treatment of the recombinant protein rPru av 1 in the absence or presence of carbohydrates, SDS-PAGE, enzyme allergosorbent tests, and inhibition assays revealed that thermal treatment of rPru av 1 alone did not show any influence on the IgE-binding activity of the protein at least for 30 min, thus correlating well with the refolding of the allergen in buffer solution as demonstrated by CD spectroscopic experiments. Incubation of the protein with starch and maltose also showed no effect on IgE-binding activity, whereas reaction with glucose and ribose and, even more pronounced, with the carbohydrate breakdown products glyceraldehyde and glyoxal induced a strong decrease of the IgE-binding capacity of rPru av 1. In the second part of the study, the effect of polyphenoloxidase-catalyzed oxidation of polyphenols on food allergen activity was investigated. Incubation of rPru av 1 with epicatechin in the presence of tyrosinase led to a drastic decrease in IgE-binding activity of the protein. Variations of the phenolic compound revealed caffeic acid and epicatechin as the most active inhibitors of the IgE-binding activity of rPru av 1, followed by catechin and gallic acid, and, finally, by quercetin and rutin, showing significantly lower activity. On the basis of these data, reactive intermediates formed during thermal carbohydrate degradation as well as during enzymatic polyphenol oxidation are suggested as the active chemical species responsible for modifying nucleophilic amino acid side chains of proteins, thus inducing an irreversible change in the tertiary structure of the protein and resulting in a loss of conformational epitopes of the allergen.

  5. 60Co-γ Irradiation Affects the Enzymatic Antioxidant System of the Citrus Red Mite Panonychus citri (Acari: Tetranychidae

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    Ke Zhang


    Full Text Available Radio-(60Co, which emits γ rays, has been used worldwide in pest control. The aim of the present study was to analyze the effect of effective-low-power 60Co-γ irradiation on the enzymatic antioxidant system of the citrus red mite Panonychus citri. One day old female adults were exposed to 0.4 kGy 60Co-γ irradiation and on the, 6th h, 1st day, 2nd day, and 5th day post treatment, the mites were euthanized for biochemical analysis. The activity of superoxide dismutase (SOD, catalase (CAT, peroxidase (POD, phenoloxidase (PO and acetylocholinesterase (AchE were investigated. POD and CAT activities, as well as SOD were higher in the irradiated mites. We found that exposure to 60Co-γ radiation resulted in increased activities of SOD, CAT, POD and decreased AchE activity. When the recovery time lasted till the 5th day, the activities of POD and PO were significantly lower than the control, whereas the SOD, CAT and AchE activities returned to control levels. Cells possess protein repair pathways to rescue oxidized proteins and restore their functions, but if these repair processes fail, oxidized proteins may become cytotoxic. Our results confirm the hypothesis that low dosages of 60Co-γ irradiation increase the level of oxidative stress in P. citri adults in a short time, causing meanwhile damage and sterility. The results of this study also indicate that stress caused by exposure to irradiation could inhibit the cholinergic system in P. citri.

  6. Factors affecting retention of early pregnancy in dairy cattle. (United States)

    Starbuck, Melanie J; Dailey, Robert A; Inskeep, E Keith


    Potential factors affecting retention of pregnancy during weeks 5-9 of gestation were studied in dairy cows and heifers (N = 211) on two farms. Cows were examined by ultrasonography for presence of a viable embryo, and sizes of the corpus luteum (CL) and of follicles > or = 5mm were recorded. Blood samples were taken at each examination and assayed for progesterone and estradiol. Overall pregnancy loss was 11.4%. Cows with two CL did not have greater concentrations of progesterone than cows with one CL and they retained fewer pregnancies (P Embryos that were lost apparently died before CL regression. Retention of pregnancy declined in cows with high body condition and as age of the cow increased. Pregnancy retention was lower in cows bred to one of four frequently-used service sires (P body condition and service sire.

  7. Perception of affective prosody in patients at an early stage of relapsing-remitting multiple sclerosis. (United States)

    Kraemer, Markus; Herold, Michele; Uekermann, Jennifer; Kis, Bernhard; Daum, Irene; Wiltfang, Jens; Berlit, Peter; Diehl, Rolf R; Abdel-Hamid, Mona


    Cognitive dysfunction is well known in patients suffering from multiple sclerosis (MS) and has been described for many years. Cognitive impairment, memory, and attention deficits seem to be features of advanced MS stages, whereas depression and emotional instability already occur in early stages of the disease. However, little is known about processing of affective prosody in patients in early stages of relapsing-remitting MS (RRMS). In this study, tests assessing attention, memory, and processing of affective prosody were administered to 25 adult patients with a diagnosis of RRMS at an early stage and to 25 healthy controls (HC). Early stages of the disease were defined as being diagnosed with RRMS in the last 2 years and having an Expanded Disability Status Scale (EDSS) of 2 or lower. Patients and HC were comparable in intelligence quotient (IQ), educational level, age, handedness, and gender. Patients with early stages of RRMS performed below the control group with respect to the subtests 'discrimination of affective prosody' and 'matching of affective prosody to facial expression' for the emotion 'angry' of the 'Tübingen Affect Battery'. These deficits were not related to executive performance. Our findings suggest that emotional prosody comprehension is deficient in young patients with early stages of RRMS. Deficits in discriminating affective prosody early in the disease may make misunderstandings and poor communication more likely. This might negatively influence interpersonal relationships and quality of life in patients with RRMS.

  8. Amygdala reactivity to sad faces in preschool children: An early neural marker of persistent negative affect

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    Michael S. Gaffrey


    Conclusions: The current findings provide preliminary evidence for amygdala activity as a potential biomarker of persistent negative affect during early childhood and suggest future work examining the origins and long-term implications of this relationship is necessary.

  9. A Preliminary Investigation of Factors Affecting Appraisal of the Decision to Take Early Retirement. (United States)

    Gowan, Mary A.


    Examines why individuals elect to take the early retirement package offered by their employer, as well as factors affecting their appraisal of that decision. Results suggest that all early retirement decisions are not voluntary. Individuals who do not wish to retire and who had lower self-esteem, fewer financial resources, and plans to continue…

  10. Solid lipid nanoparticles affect microbial colonization and enzymatic activity throughout the decomposition of alder leaves in freshwater microcosms. (United States)

    Sampaio, A C; Mendes, R J; Castro, P G; Silva, A M


    Solid lipid nanoparticles (SLNs) are used as carriers for drug delivery, and are high biocompatible and designed to endure in the host organism. Despite its current industrial production is low, many of these substances are available on the market, and much more are in the production pipeline. As a result, many of them will end in aquatic systems raising the question whether they can pose a risk to aquatic biota and the associated ecological processes. Microbial decomposers of plant litter, play a key role in forested streams being responsible for the energy flow between terrestrial and aquatic environments. Here, we investigated the effects of SLNs on alder leaf litter decomposition by aquatic microbes. Alder leaves were immersed in a stream of Northeast Portugal to allow microbial colonization before being exposed in microcosms of two types of SLNs at two concentrations for 42 days. Results showed that rates of leaf decomposition decreased with exposure to SLNs. Bacterial biomass was not inhibited by SLNs, and cultivable fungi densities remained constant (SLN-A) or increased (SLN-C) compared with control microcosms. The type and concentration of SLNs influenced differently the leaf colonization by fungi as well as fungal sporulation rate. These effects were accompanied by changes in the community extraenzymatic profile: the activities of alkaline phosphatase, acidic phosphatase, Naphthol-AS-BI-phosphohydrolase (P cycle) and lipases increased in the SLNs microcosms. This study provided the first evidence of the adverse effects of the release of SLNs to streams on leaf litter decomposition. Those effects seem to depend on the composition and concentration of SLNs, as well on the microbial target group, or enzyme. Thus, prior to massive industrial production of these nanomaterials, some measures should be taken to avoid environmental impact affecting the microbial communities responsible for detritus decomposition.


    Directory of Open Access Journals (Sweden)

    Richard G. Erskine


    Full Text Available Part 2 of a case study trilogy on early affect-confusion describes the psychotherapy relationship between an angry/helpless client and the psychotherapist’s skill in balancing the “borderline” between behavioral management and affect attunement, historical inquiry and normalization, validation and shame. The therapeutic use of bifurcated questions and juxtaposition reactions is illustrated.

  12. Early experience affects adult personality in the red junglefowl: A role for cognitive stimulation? (United States)

    Zidar, Josefina; Sorato, Enrico; Malmqvist, Ann-Marie; Jansson, Emelie; Rosher, Charlotte; Jensen, Per; Favati, Anna; Løvlie, Hanne


    Despite intense research efforts, biologists are still puzzled by the existence of animal personality. While recent studies support a link between cognition and personality, the directionality of this relationship still needs to be clarified. Early-life experiences can affect adult behaviour, and among these, cognitive stimulation has been suggested theoretically to influence personality. Yet, the influence of early cognitive stimulation has rarely been explored in empirical investigations of animal behaviour and personality. We investigated the effect of early cognitive stimulation on adult personality in the red junglefowl (Gallus gallus). To this end, we assessed adult behaviour across a number of personality assays and compared behaviour of individuals previously exposed to a series of learning tasks as chicks, with that of control individuals lacking this experience. We found that individuals exposed to early stimulation were, as adults, more vigilant and performed fewer escape attempts in personality assays. Other behaviours describing personality traits in the fowl were not affected. We conclude that our results support the hypothesis that early stimulation can affect aspects of adult behaviour and personality, suggesting a hitherto underappreciated causality link between cognition and personality. Future research should aim to confirm these findings and resolve their underlying dynamics and proximate mechanisms.

  13. The factors affecting early death after the initial therapy of acute myeloid leukemia (United States)

    Malkan, Umit Yavuz; Gunes, Gursel; Eliacik, Eylem; Haznedaroglu, Ibrahim Celalettin; Etgul, Sezgin; Aslan, Tuncay; Yayar, Okan; Aydin, Seda; Demiroglu, Haluk; Ozcebe, Osman Ilhami; Sayinalp, Nilgun; Goker, Hakan; Aksu, Salih; Buyukasik, Yahya


    There are some improvements in management of acute myeloid leukemia (AML). However, induction-induced deaths still remain as a major problem. The aim of this study is to assess clinical parameters affecting early death in patients with AML. 199 AML patients, who were treated with intensive, non-intensive or supportive treatment between 2002 and 2014 in Hacettepe Hematology Department, were analyzed retrospectively. In our study early death rate for elderly was found to be lower than previous reports whereas it was similar for those who were under age of 60. Better ECOG performance (ECOG performance score 0 and 1) and non-intensive treatment associated with lower early death rates, however APL-type disease associated with higher early death rates. ECOG performance score at diagnosis was found to be the most related independent factor with higher rate of early death in 15 days after treatment (P<0.001). Therefore we decided to understand the factors which were related with ECOG. WBC count at diagnosis was found to be the only related parameter with ECOG performance score. Leucocyte count at diagnosis appears like to have an indirect effect on early death in AML patients. It maybe suggested that in recent years there is an improvement in early death rates of elderly AML patients. The currently reported findings require prospective validation and would encourage the incorporation of other next generation genomics for the prediction of early death and overall risk status of AML. PMID:26885243

  14. Early Negative Affect Predicts Anxiety, Not Autism, in Preschool Boys with Fragile X Syndrome (United States)

    Tonnsen, Bridgette L.; Malone, Patrick S.; Hatton, Deborah D.; Roberts, Jane E.


    Children with fragile X syndrome (FXS) face high risk for anxiety disorders, yet no studies have explored FXS as a high-risk sample for investigating early manifestations of anxiety outcomes. Negative affect is one of the most salient predictors of problem behaviors and has been associated with both anxiety and autistic outcomes in clinical and…

  15. How does early detection by screening affect disease progression?: Modeling estimated benefits in prostate cancer screening

    NARCIS (Netherlands)

    E.M. Wever (Elisabeth); G. Draisma (Gerrit); E.A.M. Heijnsdijk (Eveline); H.J. de Koning (Harry)


    textabstractBackground. Simulation models are essential tools for estimating benefits of cancer screening programs. Such models include a screening-effect model that represents how early detection by screening followed by treatment affects disease-specific survival. Two commonly used screening-effec

  16. Factors Affecting Early Services for Children Who Are Hard of Hearing (United States)

    Harrison, Melody; Page, Thomas A.; Oleson, Jacob; Spratford, Meredith; Berry, Lauren Unflat; Peterson, Barbara; Welhaven, Anne; Arenas, Richard M.; Moeller, Mary Pat


    Purpose: To describe factors affecting early intervention (EI) for children who are hard of hearing, we analyzed (a) service setting(s) and the relationship of setting to families' frequency of participation, and (b) provider preparation, caseload composition, and experience in relation to comfort with skills that support spoken language for…

  17. Timing of examinations affects school performance differently in early and late chronotypes

    NARCIS (Netherlands)

    van der Vinne, Vincent; Zerbini, Giulia; Siersema, Anne; Pieper, Amy; Merrow, Martha; Hut, Roelof A.; Roenneberg, Till; Kantermann, Thomas


    Circadian clocks of adolescents typically run lateincluding sleep timesyet adolescents generally are expected at school early in the morning. Due to this mismatch between internal (circadian) and external (social) times, adolescents suffer from chronic sleep deficiency, which, in turn, affects acade

  18. Non-enzymatic modifications of prostaglandin H synthase 1 affect bifunctional enzyme activity - Implications for the sensitivity of blood platelets to acetylsalicylic acid. (United States)

    Kassassir, Hassan; Siewiera, Karolina; Talar, Marcin; Stec-Martyna, Emilia; Pawlowska, Zofia; Watala, Cezary


    Due to its ability to inhibit the blood platelet PGHS-1, acetylsalicylic acid (ASA, Aspirin(®)) is widely used as a preventive agent in atherothrombotic diseases. However, its beneficial effects seem to be lower in diabetic patients, suggesting that protein glycation may impair effective ASA-mediated acetylation process. On the other hand, it is proposed that ASA can prevent some of the late complications of diabetes by lowering the extent of glycation at protein free amino groups. The aim of this work was to evaluate the extents of non-enzymatic N-glycosylation (glycation) and acetylation of blood platelet PGHS-1 (COX-1) and the competition between glycation and acetylation was investigated in order to demonstrate how these two reactions may compete against platelet PGHS-1. When PGHS-1 was incubated with glycating/acetylating agents (glucose, Glu; 1,6-bisphosphofructose, 1,6-BPF; methylglyoxal, MGO, acetylsalicylic acid, ASA), the enzyme was modified in 13.4 ± 1.6, 5.3 ± 0.5, 10.7 ± 1.2 and 6.4 ± 1.1 mol/mol protein, respectively, and its activity was significantly reduced. The prior glycation/carbonylation of PGHS-1 with Glu, 1,6-BPF or MGO decreased the extent of acetylation from 6.4 ± 1.1 down to 2.5 ± 0.2, 3.6 ± 0.3 and 5.2 ± 0.2 mol/mol protein, respectively, but the enzyme still remained susceptible to the subsequent inhibition of its activity with ASA. When PGHS-1 was first acetylated with ASA and then incubated with glycating/carbonylating agents, we observed the following reductions in the enzyme modifications: from 13.4 ± 1.6 to 8.7 ± 0.6 mol/mol protein for Glu, from 5.3 ± 0.5 to 3.9 ± 0.3 mol/mol protein for 1,6-BPF and from 10.7 ± 1.2 to 7.5 ± 0.5 mol/mol protein for MGO, however subsequent glycation/carbonylation did not significantly affect PGHS-1 function. Overall, our outcomes allow to better understand the structural aspects of the chemical competition between glycation and acetylation of PGHS-1.

  19. Early Negative Affect Predicts Anxiety, not Autism, in Preschool Boys with Fragile X Syndrome


    Tonnsen, Bridgette L.; Malone, Patrick S.; Hatton, Deborah D.; Roberts, Jane E.


    Children with fragile X syndrome (FXS) face high risk for anxiety disorders, yet no studies have explored FXS as a high-risk sample for investigating early manifestations of anxiety outcomes. Negative affect is one of the most salient predictors of problem behaviors and has been associated with both anxiety and autistic outcomes in clinical and non-clinical pediatric samples. In light of the high comorbidity between autism and anxiety within FXS, the present study investigates the relationshi...

  20. Frontal lobe changes occur early in the course of affective disorders in young people

    Directory of Open Access Journals (Sweden)

    Lagopoulos Jim


    Full Text Available Abstract Background More severe and persistent forms of affective disorders are accompanied by grey matter loss in key frontal and temporal structures. It is unclear whether such changes precede the onset of illness, occur early in the course or develop gradually with persistence or recurrence of illness. A total of 47 young people presenting with admixtures of depressive and psychotic symptoms were recruited from specialist early intervention services along with 33 age matched healthy control subjects. All participants underwent magnetic resonance imaging and patients were rated clinically as to current stage of illness. Twenty-three patients were identified as being at an early 'attenuated syndrome' stage, while the remaining were rated as having already reached the 'discrete disorder' or 'persistent or recurrent illness' stage. Contrasts were carried out between controls subjects and patients cohorts with attenuated syndromes and discrete disorders, separately. Results The patients that were identified as having a discrete or persisting disorder demonstrated decreased grey matter volumes within distributed frontal brain regions when contrasted to both the control subjects as well as those patients in the attenuated syndrome stage. Overall, patients who were diagnosed as more advanced in terms of the clinical stage of their illness, exhibited the greatest grey matter volume loss of all groups. Conclusions This study suggests that, in terms of frontal grey matter changes, a major transition point may occur in the course of affective illness between early attenuated syndromes and later discrete illness stages.

  1. Early life antibiotic exposure affects pancreatic islet development and metabolic regulation (United States)

    Li, Jiaying; Yang, Kaiyuan; Ju, Tingting; Ho, Tracy; McKay, Catharine A.; Gao, Yanhua; Forget, Shay K.; Gartner, Stephanie R.; Field, Catherine J.; Chan, Catherine B.; Willing, Benjamin P.


    Childhood antibiotic exposure has been recently linked with increased risk of metabolic disease later in life. A better understanding of this association would potentially provide strategies to reduce the childhood chronic disease epidemic. Therefore, we explored the underlying mechanisms using a swine model that better mimics human infants than rodents, and demonstrated that early life antibiotic exposure affects glucose metabolism 5 weeks after antibiotic withdrawal, which was associated with changes in pancreatic development. Antibiotics exerted a transient impact on postnatal gut microbiota colonization and microbial metabolite production, yet changes in the expression of key genes involved in short-chain fatty acid signaling and pancreatic development were detected in later life. These findings suggest a programming effect of early life antibiotic exposure that merits further investigation. PMID:28150721

  2. Adjustments in CAM and enzymatic scavenging of H2O2 in juvenile plants of the epiphytic bromeliad Guzmania monostachia as affected by drought and rewatering. (United States)

    Carvalho, Victória; Abreu, Maria E; Mercier, Helenice; Nievola, Catarina C


    Juvenile plants of epiphytes such as bromeliads are highly prone to dehydration under drought conditions. It is likely that young epiphytes evolved mostly metabolic strategies to resist drought, which may include the plastic modulation of the enzymatic antioxidant system and crassulacean acid metabolism (CAM). Few studies have investigated such strategies in juvenile epiphytes, although such research is important to understand how these plants might face drought intensification derived from potential climatic alterations. The epiphytic CAM bromeliad Guzmania monostachia (L.) Rusby ex Mez var. monostachia is known to have plastic responses to drought, but no reports have focused on the metabolism of juvenile plants to drought and recovery. Hence, we aimed to verify how juvenile G. monostachia plants adjust malate (indicative of CAM), H2O2 content and enzymatic scavenging in response to drought (eight days without irrigation) and rewatering (six days of irrigation post-drought). Interestingly, drought decreased H2O2 content and activities of superoxide dismutase, catalase (CAT) and ascorbate peroxidase (APX) in the pre-dusk period, although glutathione reductase (GR) and CAM activity increased. Rewatering restored H2O2, but activities of APX, CAT and GR exceeded pre-stress levels in the pre-dusk and/or pre-dawn periods. Results suggest that recovery from a first drought redefines the homeostatic balance of H2O2 scavenging, in which rewatered plants stimulate the enzymatic antioxidant system while drought-exposed plants intensify CAM activity to regulate H2O2 content, a photosynthetic pathway known to prevent oxidative stress. Such data show that young G. monostachia plants adjust CAM and H2O2 scavenging to adapt to water availability.

  3. Modeling naturalistic craving, withdrawal, and affect during early nicotine abstinence: A pilot ecological momentary assessment study. (United States)

    Bujarski, Spencer; Roche, Daniel J O; Sheets, Erin S; Krull, Jennifer L; Guzman, Iris; Ray, Lara A


    Despite the critical role of withdrawal, craving, and positive affect (PA) and negative affect (NA) in smoking relapse, relatively little is known about the temporal and predictive relationship between these constructs within the first day of abstinence. This pilot study aims to characterize dynamic changes in withdrawal, craving, and affect over the course of early abstinence using ecological momentary assessment. Beginning immediately after smoking, moderate and heavy smoking participants (n = 15 per group) responded to hourly surveys assessing craving, withdrawal, NA, and PA. Univariate and multivariate multilevel random coefficient modeling was used to describe the progression of craving, withdrawal/NA, and PA and to test correlations between these constructs at the subject level over the course of early abstinence. Heavy smokers reported greater craving from 1-4 hr of abstinence and greater withdrawal/NA after 3 or more hours as compared with moderate smokers. Level of withdrawal/NA was strongly positively associated with craving, and PA was negatively correlated with craving; however, the temporal dynamics of these correlations differed substantially. The association between withdrawal/NA and craving decreased over early abstinence, whereas the reverse was observed for PA. These findings can inform experimental studies of nicotine abstinence as well as their clinical applications to smoking cessation efforts. In particular, these results help to elucidate the role of PA in nicotine abstinence by demonstrating its independent association with nicotine craving over and above withdrawal/NA. If supported by future studies, these findings can refine experimental methods and clinical approaches for smoking cessation.

  4. Early life trauma is associated with altered white matter integrity and affective control. (United States)

    Corbo, Vincent; Amick, Melissa A; Milberg, William P; McGlinchey, Regina E; Salat, David H


    Early life trauma (ELT) has been shown to impair affective control and attention well into adulthood. Neuroimaging studies have further shown that ELT was associated with decreased white matter integrity in the prefrontal areas in children and adults. However, no study to date has looked at the relationship between white matter integrity and affective control in individuals with and without a history of ELT. To examine this, we tested 240 Veterans with (ELT N = 80) and without (NoELT N = 160) a history of childhood sexual abuse, physical abuse or family violence. Affective control was measured with the Affective Go/No-Go (AGN) and attention was indexed with the Test of Variable Attention (TOVA). White matter integrity was measured using fractional anisotropy (FA). Results showed greater number of errors on the AGN in ELT compared to NoELT. There was no difference on the TOVA. While there were no mean differences in FA, there was an interaction between FA and reaction time to positive stimuli on the AGN where the ELT group showed a positive relationship between FA and reaction time in right frontal and prefrontal areas, whereas the NoELT group showed a negative or no association between FA and reaction time. This suggests that ELT may be associated with a distinct brain-behavior relationship that could be related to other determinants of FA than those present in healthy adults.

  5. Does classroom separation affect twins' reading ability in the early years of school? (United States)

    Coventry, William L; Byrne, Brian; Coleman, Marreta; Olson, Richard K; Corley, Robin; Willcutt, Eric; Samuelsson, Stefan


    In this article we report on reading ability of twin children in kindergarten to Grade 2 as a function of whether members of the pairs are assigned to the same or different classrooms. All analyses were run using mixed model regressions to account for the interdependence between twin pairs. The samples, total N = 1505, are from Australia and the United States. We found a close-to-significant difference in favor of same-class children in kindergarten and Grade 1. However, when results were adjusted to take account of pre-existing differences in disruptive behavior and in preliteracy ability, the class assignment effects disappeared. We suggest that these pre-existing differences, particularly disruptive behavior, are influencing decisions about whether to separate twins or not and also affecting early reading performance, a conclusion supported by significant correlations between the behavioral measures, preliteracy, and school-based reading. We conclude that, on average, early literacy in twins is not directly affected by their assignment to the same or different classrooms.

  6. Early developmental conditions affect stress response in juvenile but not in adult house sparrows (Passer domesticus). (United States)

    Lendvai, Adám Z; Loiseau, Claire; Sorci, Gabriele; Chastel, Olivier


    The short- and long-term consequences of developmental conditions on fitness have received growing attention because the environmental conditions during early life may influence growth, condition at independence, recruitment, reproductive success or survival. We tested here, in a natural house sparrow population, if early conditions during nestling stage affected the stress response of the birds (i) shortly after fledging and (ii) next year, during their first breeding. We experimentally manipulated brood size to mimic different rearing conditions, creating reduced (-2 chicks) and enlarged broods (+2 chicks), while in a third group brood size was not manipulated. Nestling nutrition state decreased with post-manipulation brood sizes as indicated by lower body mass. Fledglings with higher body mass at the age of ten days showed lower stress response than birds that were leaner at the same age. Fledglings reared in large broods showed a higher response to stress protocol than chicks from small broods, and this effect was in significant interaction with the age of fledglings at capture. This interaction indicates that the effects of the brood size became gradually smaller as the fledglings grew older and were further from their nestling period. The effects of early conditions vanished by the next year: the stress response of adult first time breeders was unrelated to the brood size they fledged from. These results suggest that stress response may reflect the actual state of an individual, rather than its developmental history.

  7. The early social environment affects social competence in a cooperative breeder (United States)

    Taborsky, Barbara; Arnold, Cornelia; Junker, Julian; Tschopp, Andreas


    Social competence is defined as the ability of an animal to optimize the expression of social behaviour as a function of the available social information. The social environment encountered early in life can affect the expression of various social behaviours later in life. We investigated whether early social experience can affect social competence. In the cooperatively breeding cichlid Neolamprologus pulcher, we tested whether individuals reared with older brood-caring conspecifics persistently perform better in a series of tasks (1) simulating different social contexts, (2) assigning individuals different social roles and (3) exposing them to an unknown social situation. Fish that had been reared together with older conspecifics showed more appropriate behaviours both as winners (more aggressive displays) and as losers (more submissive displays) when aggressively competing with peers over a resource, and when trying to be accepted as subordinate group member and prospective brood care helper by an unfamiliar dominant pair (more submissive displays near shelters), a situation they had never encountered before. In both tasks fish that had grown up with older fish were tolerated better by conspecifics than fish reared with same-age siblings only. We detected effects of the early environment on social behaviour in the juvenile and adult stages of the test fish. Our results suggest that growing up in more complex social groups fosters a general social ability (i.e. social competence) in N. pulcher that improves their performance across different social roles and contexts, and which may provide fitness benefits. PMID:22536004

  8. Moderate recurrent hypoglycemia during early development leads to persistent changes in affective behavior in the rat. (United States)

    Moore, Holly; Craft, Tara K S; Grimaldi, Lisa M; Babic, Bruna; Brunelli, Susan A; Vannucci, Susan J


    Recurrent hypoglycemia is a common problem among infants and children that is associated with several metabolic disorders and insulin-dependent diabetes mellitus. Although studies have reported a relationship between a history of juvenile hypoglycemia and psychological health problems, the direct effects of recurrent moderate hypoglycemia have not been fully determined. Thus, in this study, we used an animal model to examine the effects of recurrent hypoglycemia during the juvenile period on affective, social, and motor function (assessed under euglycemic conditions) across development. To model recurrent hypoglycemia, rats were administered 5 U/kg of insulin or saline twice per day from postnatal day (P)10 to P19. Body weight gain was retarded in insulin-treated rats during the treatment period, but recovered by the end of treatment. However, insulin-treated rats displayed increases in affective reactivity that emerged early during treatment and persisted after treatment into early adulthood. Specifically, insulin-treated pups showed increased maternal separation-induced vocalizations as infants, and an exaggerated acoustic startle reflex as juveniles and young adults. Moreover, young adult rats with a history of recurrent juvenile hypoglycemia exhibited increased fear-potentiated startle and increases in behavioral and hormonal responses to restraint stress. Some of these effects were sex-dependent. The changes in affective behavior in insulin-exposed pups were accompanied by decreases in adolescent social play behavior. These results provide evidence that recurrent, transient hypoglycemia during juvenile development can lead to increases in fear-related behavior and stress reactivity. Importantly, these phenotypes are not reversed with normalization of blood glucose and may persist into adulthood.

  9. Parents and early life environment affect behavioral development of laying hen chickens. (United States)

    de Haas, Elske N; Bolhuis, J Elizabeth; Kemp, Bas; Groothuis, Ton G G; Rodenburg, T Bas


    Severe feather pecking (SFP) in commercial laying hens is a maladaptive behavior which is associated with anxiety traits. Many experimental studies have shown that stress in the parents can affect anxiety in the offspring, but until now these effects have been neglected in addressing the problem of SFP in commercially kept laying hens. We therefore studied whether parental stock (PS) affected the development of SFP and anxiety in their offspring. We used flocks from a brown and white genetic hybrid because genetic background can affect SFP and anxiety. As SFP can also be influenced by housing conditions on the rearing farm, we included effects of housing system and litter availability in the analysis. Forty-seven rearing flocks, originating from ten PS flocks were followed. Behavioral and physiological parameters related to anxiety and SFP were studied in the PS at 40 weeks of age and in the rearing flocks at one, five, ten and fifteen weeks of age. We found that PS had an effect on SFP at one week of age and on anxiety at one and five weeks of age. In the white hybrid, but not in the brown hybrid, high levels of maternal corticosterone, maternal feather damage and maternal whole-blood serotonin levels showed positive relations with offsprings' SFP at one week and offsprings' anxiety at one and five weeks of age. Disruption and limitation of litter supply at an early age on the rearing farms increased SFP, feather damage and fearfulness. These effects were most prominent in the brown hybrid. It appeared that hens from a brown hybrid are more affected by environmental conditions, while hens from a white hybrid were more strongly affected by parental effects. These results are important for designing measures to prevent the development of SFP, which may require a different approach in brown and white flocks.

  10. Application and analysis of debris-flow early warning system in Wenchuan earthquake-affected area (United States)

    Liu, D. L.; Zhang, S. J.; Yang, H. J.; Zhao, L. Q.; Jiang, Y. H.; Tang, D.; Leng, X. P.


    The activities of debris flow (DF) in the Wenchuan earthquake-affected area significantly increased after the earthquake on 12 May 2008. The safety of the lives and property of local people is threatened by DFs. A physics-based early warning system (EWS) for DF forecasting was developed and applied in this earthquake area. This paper introduces an application of the system in the Wenchuan earthquake-affected area and analyzes the prediction results via a comparison to the DF events triggered by the strong rainfall events reported by the local government. The prediction accuracy and efficiency was first compared with a contribution-factor-based system currently used by the weather bureau of Sichuan province. The storm on 17 August 2012 was used as a case study for this comparison. The comparison shows that the false negative rate and false positive rate of the new system is, respectively, 19 and 21 % lower than the system based on the contribution factors. Consequently, the prediction accuracy is obviously higher than the system based on the contribution factors with a higher operational efficiency. On the invitation of the weather bureau of Sichuan province, the authors upgraded their prediction system of DF by using this new system before the monsoon of Wenchuan earthquake-affected area in 2013. Two prediction cases on 9 July 2013 and 10 July 2014 were chosen to further demonstrate that the new EWS has high stability, efficiency, and prediction accuracy.

  11. Distraction by emotion in early adolescence: affective facilitation and interference during the attentional blink

    Directory of Open Access Journals (Sweden)

    Sabine eHeim


    Full Text Available This study examined the extent to which early adolescents (aged 10 to 13 years differ from adults in their sensitivity to attention capture by affective stimuli during rapid processing. A rapid serial visual presentation paradigm (RSVP was implemented as a dual task, requiring the report of two green target stimuli embedded in a stream of distractors. Known as the attentional blink (AB, task performance is typically impaired when the first and second targets (T1 and T2, respectively are separated by at least one distractor and about 200 ms of time. Here we used written verbs of pleasant, neutral, and unpleasant content as T1 items, while affectively neutral exemplars served as T2 and distractor events. The temporal distance between T1 and T2 was manipulated to contain either one distractor (intertarget interval 232 ms or five distractors (intertarget interval 696 ms. Students reported pleasant T1 words more accurately, compared to neutral and unpleasant words, indicating facilitation of appetitive content on performance during RSVP. Emotional relevance of T1 was at the expense of T2 accuracy: At an intertarget interval of 232 ms (i.e., during the AB period, identification of (neutral T2 words was impaired when preceded by pleasant and unpleasant T1s. No interference across targets was observed, however, beyond the blink period, in which T1 and T2 were separated by 696 ms. Thus, emotionally relevant events capture and hold attentional resources, at the cost of attentive processing in subsequent episodes. Contrary to our findings in adults, these capture effects were most obvious when the available capacity was limited, i.e., during the critical interval of the AB. The findings are discussed in light of the use of alternative cognitive strategies as development proceeds beyond early adolescence into adulthood.

  12. Early life in a barren environment adversely affects spatial cognition in laying hens (Gallus gallus domesticus

    Directory of Open Access Journals (Sweden)

    Fernanda Machado Tahamtani


    Full Text Available Spatial cognition in vertebrates is adversely affected by a lack of environmental complexity during early life. However, to our knowledge no previous studies have tested the effect of early exposure to varying degrees of environmental complexity on specific components of spatial cognition in chickens. There are two main rearing systems for laying hens in the EU: aviaries and cages. These two systems differ from one another in environmental complexity. The aim of the present study was to test the hypothesis that rearing in a barren cage environment relative to a complex aviary environment causes long-lasting deficits in the ability to perform spatial tasks. For this purpose, 24 white Dekalb laying hens, half of which had been reared in an aviary system and the other half in a conventional cage system, were tested in a holeboard task. Birds from both treatment groups learnt the task, however the cage-reared hens required more time to locate rewards and had poorer levels of working memory. The latter finding supports the hypothesis that rearing in a barren environment causes long-term impairment of short-term memory in chickens.

  13. Modifications on the hydrogen bond network by mutations of Escherichia coli copper efflux oxidase affect the process of proton transfer to dioxygen leading to alterations of enzymatic activities

    Energy Technology Data Exchange (ETDEWEB)

    Kajikawa, Takao; Kataoka, Kunishige [Graduate School of Natural Science and Technology, Kanazawa University, Kakuma, Kanazawa 920-1192 (Japan); Sakurai, Takeshi, E-mail: [Graduate School of Natural Science and Technology, Kanazawa University, Kakuma, Kanazawa 920-1192 (Japan)


    Highlights: Black-Right-Pointing-Pointer Proton transfer pathway to dioxygen in CueO was identified. Black-Right-Pointing-Pointer Glu506 is the key amino acid to transport proton. Black-Right-Pointing-Pointer The Ala mutation at Glu506 formed a compensatory proton transfer pathway. Black-Right-Pointing-Pointer The Ile mutation at Glu506 shut down the hydrogen bond network. -- Abstract: CueO has a branched hydrogen bond network leading from the exterior of the protein molecule to the trinuclear copper center. This network transports protons in the four-electron reduction of dioxygen. We replaced the acidic Glu506 and Asp507 residues with the charged and uncharged amino acid residues. Peculiar changes in the enzyme activity of the mutants relative to the native enzyme indicate that an acidic amino acid residue at position 506 is essential for effective proton transport. The Ala mutation resulted in the formation of a compensatory hydrogen bond network with one or two extra water molecules. On the other hand, the Ile mutation resulted in the complete shutdown of the hydrogen bond network leading to loss of enzymatic activities of CueO. In contrast, the hydrogen bond network without the proton transport function was constructed by the Gln mutation. These results exerted on the hydrogen bond network in CueO are discussed in comparison with proton transfers in cytochrome oxidase.

  14. Early developmental gene enhancers affect subcortical volumes in the adult human brain. (United States)

    Becker, Martin; Guadalupe, Tulio; Franke, Barbara; Hibar, Derrek P; Renteria, Miguel E; Stein, Jason L; Thompson, Paul M; Francks, Clyde; Vernes, Sonja C; Fisher, Simon E


    Genome-wide association screens aim to identify common genetic variants contributing to the phenotypic variability of complex traits, such as human height or brain morphology. The identified genetic variants are mostly within noncoding genomic regions and the biology of the genotype-phenotype association typically remains unclear. In this article, we propose a complementary targeted strategy to reveal the genetic underpinnings of variability in subcortical brain volumes, by specifically selecting genomic loci that are experimentally validated forebrain enhancers, active in early embryonic development. We hypothesized that genetic variation within these enhancers may affect the development and ultimately the structure of subcortical brain regions in adults. We tested whether variants in forebrain enhancer regions showed an overall enrichment of association with volumetric variation in subcortical structures of >13,000 healthy adults. We observed significant enrichment of genomic loci that affect the volume of the hippocampus within forebrain enhancers (empirical P = 0.0015), a finding which robustly passed the adjusted threshold for testing of multiple brain phenotypes (cutoff of P Brain Mapp 37:1788-1800, 2016. © 2016 Wiley Periodicals, Inc.

  15. Plant Spacing and Cultivar Affects Yield Components, Qualitative Traits and Early Ripening of Tomato (Lycopersicon esculentum

    Directory of Open Access Journals (Sweden)

    Ebrahim MAMNOIE


    Full Text Available Two-year field trial was set up on sandy clay soil in the Jiroft and Kahnouj Agricultural Research Center with the objective to determine the effect of plant spacing and different cultivars on the yield and qualitative characteristics of tomato (Lycopersicon esculentum. This experiment was performed as split plot based on complete randomized block design with 3 replications. The main plots were in – row spacing in 4 levels include 0.3, 0.4, 0.5 and 0.6 m and the subplots were cultivars naming ‘Shef ’, ‘Peto Early CH’ and ‘FDT 202’. The results showed that, fruit length to diameter ratio, total acidity, fruit number per plant, mean fruit weight, yield per plant, total yield and first harvest to total harvests ratio were significantly affected by plant spacing. In addition, cultivars showed significant effect on all traits evaluated (p<0.01. In this experiment, fruit length to diameter ratio and total acidity increased as plant spacing increased, however it had no effect on total soluble solids. In this study, total fruit yield is being increased while the yield per plant, number of fruit per plant and fruit weight is being reduced by increased number of plants per unit area. Although among tomato cultivar, ‘Peto Early CH’ had a higher yield over other cultivars, but cultivar ‘Shef ’ showed higher yield in the first harvest. Generally it seems according to the results collected that plant spacing 0.3 m and ‘Shef ’ cultivar owing to better adaptation and higher commercial yield for production in Jiroft city is suggested.

  16. Early and Midterm Results Following Interventional Coarctoplasty: Evaluation of Variables that Can Affect the Results (United States)

    Bassiri, Hossein Ali; Shafe, Omid; Sarpooshi, Javad


    Background and Objectives Stent coarctoplasty has been approved as the treatment of choice for adult patients with coarctation of the aorta. We have evaluated the early and midterm clinical and procedural results after interventional coarctoplasty. Also, variables that can affect these results were evaluated. Subjects and Methods Gathering clinical, angiographic and procedural data, we evaluated the pre-specified outcomes, including procedural success, complications, the incidence of hypertension after coarctoplasty etc., after the procedure. The effect of pre-specified variables including aortic arch shape, coarctation type and etc. on the procedural result was evaluated. Results Between February 2005 through March 2014, 133 stent coarctoplasty procedures were performed. Median age was 23.5 years old (interquartile range [IQR]:19-28), and 105 (71.9%) were male. Nearly all of the patients were undergone stent coarctoplasty, mostly with cheatham platinum (CP) stents. There was no association between aortic arch morphology and acute procedural complications. Balloon length more than 40 mm (p=0.028), aorta diameter at the site of Coarctation larger than 2.35 mm (p=0.008) was associated with higher rate of restenosis during follow-up. Comparison between the prevalence of hypertension (HTN) before and after coarctoplasty showed a significant reduction in the prevalence of HTN (117 [91.4%] vs. 95 [74.2%] p<0.001). Conclusion Stent coarctoplasty is a low-risk procedure with favorable early and delayed outcomes. Most mortality is related to the patient's comorbid conditions and not to the procedure. PMID:28154597

  17. Prospective associations between forms and functions of aggression and social and affective processes during early childhood. (United States)

    Ostrov, Jamie M; Murray-Close, Dianna; Godleski, Stephanie A; Hart, Emily J


    The central goal of this study was to examine the prospective associations between forms (i.e., physical and relational) and functions (i.e., proactive and reactive) of aggressive behavior with social (i.e., peer rejection) and affective (i.e., anger, emotion regulation skills) processes during early childhood (N = 96, mean age = 42.80 months, SD = 7.57). A cross-lagged path analysis revealed that proactive relational aggression was uniquely associated with decreases in peer rejection, whereas reactive relational aggression was associated with increases in peer rejection over time. Proactive relational aggression predicted decreases in anger, whereas reactive relational aggression tended to be associated with increases in anger. Proactive relational aggression uniquely predicted increases in emotion regulation skills, whereas reactive relational aggression tended to be associated with decreases in emotion regulation skills over time. Finally, anger was significantly associated with increases in several subtypes of aggressive behavior. In sum, the findings provide further support for the distinction between subtypes of aggressive behavior in young children.

  18. Tadpoles of Early Breeding Amphibians are Negatively Affected by Leaf Litter From Invasive Chinese Tallow Trees (United States)

    Leonard, N. E.


    As wetlands are invaded by Chinese tallow trees (Triadica sebifera), native trees are displaced and detrital inputs to amphibian breeding ponds are altered. I used a mesocosm experiment to examine the effect of Chinese tallow leaf litter on the survival to, size at, and time to metamorphosis of amphibian larvae. Fifty 1000-L cattle watering tanks were treated with 1500 g dry weight of one of five leaf litter treatments: Chinese tallow, laurel oak (Quercus laurifolia), water tupelo (Nyssa aquatica), slash pine (Pinus elliottii), or a 3:1:1:1 mixture. Each tank received 45 tadpoles of Pseudacris feriarum, Bufo terrestris, and Hyla cinerea in sequence according to their natural breeding phonologies. Every Pseudacris feriarum and Bufo terrestris tadpole exposed to Chinese tallow died prior to metamorphosis. Hyla cinerea survival in tanks with tallow-only was significantly lower than that observed for all other leaf treatments. Hyla cinerea tadpoles from tallow-only and mixed-leaf treatments were larger at metamorphosis and transformed faster than those in tanks with native leaves only. These results suggest that Chinese tallow leaf litter may negatively affect tadpoles of early breeding frogs and that Chinese tallow invasion may change the structure of amphibian communities in temporary ponds.

  19. Carbon and nitrogen dynamics in early stages of forest litter decomposition as affected by nitrogen addition

    Institute of Scientific and Technical Information of China (English)

    DENG Xiao-wen; LIU Ying; HAN Shi-jie


    The effects of nitrogen (N) availability and tree species on the dynamics of carbon and nitrogen at early stage of decomposition of forest litter were studied in a 13-week laboratory incubation experiment. Fresh litter samples including needle litter (Pinus koraiensis) and two types of broadleaf litters (Quercus mongolica and Tilia amurensis) were collected from a broadleaf-korean pine mixed forest in the northern slope of Changbai Mountain (China). Different doses of N (equal to 0, 30 and 50 kg·ha-1yr-1, respectively, as NH4NO3) were added to litter during the experiment period. The litter decomposition rate expressed as mass loss and respiration rate increased significantly with increasing N availability. The mass loss and cumulative CO2-C emission were higher in leaf litter compared to that in needle litter. The dissolved organic Carbon (DOC) concentrations in litter leachate varied widely between the species, but were not greatly affected by N treatments. Regardless of the N addition rate, both N treatments and species had no significant effect on dissolved organic N (DON) concentrations in litter leachate. About 52·78% of added N was retained in the litter. The percentage of N retention was positively correlated (R2=0.91, p<0.05) with the litter mass loss. This suggested that a forest floor with easily decomposed litter might have higher potential N sink strength than that with more slowly decomposed litter.

  20. Transplacental and early life exposure to inorganic arsenic affected development and behavior in offspring rats

    Energy Technology Data Exchange (ETDEWEB)

    Xi, Shuhua; Jin, Yaping; Sun, Guifan [China Medical University, Department of Environmental and Occupational Health, College of Public Health, Shenyang, Liaoning (China); Sun, Wenjuan; Wang, Fengzhi [Shenyang Medical College, Department of Preventive Medicine, Shenyang, Liaoning (China)


    To evaluate the developmental neurotoxicity of arsenic in offspring rats by transplacental and early life exposure to sodium arsenite in drinking water, the pregnant rats or lactating dams, and weaned pups were given free access to drinking water, which contained arsenic at concentrations of 0, 10, 50, 100 mg/L from GD 6 until PND 42. A battery of physical and behavioral tests was applied to evaluate the functional outcome of pups. Pups in arsenic exposed groups weighed less than controls throughout lactation and weaning. Body weight of 10, 50 and 100 mg/L arsenic exposed groups decreased significantly on PND 42, 16 and 12, respectively. Physical development (pinna unfolding, fur appearance, incisor eruption, or eye opening) in pups displayed no significant differences between control and arsenic treated groups. The number of incidences within the 100 mg/L arsenic treated group, in tail hung, auditory startle and visual placing showed significant decrease compared to the control group (p<0.05). In square water maze test, the trained numbers to finish the trials successfully in 50 and 100 mg/L arsenic exposed groups increased remarkably compared to control group, and there was a dose-related increase (p<0.01) observed. Taken together, these data show that exposure of inorganic arsenite to pregnant dams and offspring pups at levels up to 100 mg/L in drinking water may affect their learning and memory functions and neuromotor reflex. (orig.)

  1. Early infancy microbial and metabolic alterations affect risk of childhood asthma. (United States)

    Arrieta, Marie-Claire; Stiemsma, Leah T; Dimitriu, Pedro A; Thorson, Lisa; Russell, Shannon; Yurist-Doutsch, Sophie; Kuzeljevic, Boris; Gold, Matthew J; Britton, Heidi M; Lefebvre, Diana L; Subbarao, Padmaja; Mandhane, Piush; Becker, Allan; McNagny, Kelly M; Sears, Malcolm R; Kollmann, Tobias; Mohn, William W; Turvey, Stuart E; Finlay, B Brett


    Asthma is the most prevalent pediatric chronic disease and affects more than 300 million people worldwide. Recent evidence in mice has identified a "critical window" early in life where gut microbial changes (dysbiosis) are most influential in experimental asthma. However, current research has yet to establish whether these changes precede or are involved in human asthma. We compared the gut microbiota of 319 subjects enrolled in the Canadian Healthy Infant Longitudinal Development (CHILD) Study, and show that infants at risk of asthma exhibited transient gut microbial dysbiosis during the first 100 days of life. The relative abundance of the bacterial genera Lachnospira, Veillonella, Faecalibacterium, and Rothia was significantly decreased in children at risk of asthma. This reduction in bacterial taxa was accompanied by reduced levels of fecal acetate and dysregulation of enterohepatic metabolites. Inoculation of germ-free mice with these four bacterial taxa ameliorated airway inflammation in their adult progeny, demonstrating a causal role of these bacterial taxa in averting asthma development. These results enhance the potential for future microbe-based diagnostics and therapies, potentially in the form of probiotics, to prevent the development of asthma and other related allergic diseases in children.

  2. Brain-derived neurotrophic factor Val66Met polymorphism and early life adversity affect hippocampal volume. (United States)

    Carballedo, Angela; Morris, Derek; Zill, Peter; Fahey, Ciara; Reinhold, Elena; Meisenzahl, Eva; Bondy, Brigitta; Gill, Michael; Möller, Hans-Jürgen; Frodl, Thomas


    The interaction between adverse life events during childhood and genetic factors is associated with a higher risk to develop major depressive disorder (MDD). One of the polymorphisms found to be associated with MDD is the Val66MET polymorphism of brain-derived neurotrophic factor (BDNF). The aim of our two-center study was to determine how the BDNF Val66Met polymorphism and childhood adversity affect the volumetric measures of the hippocampus in healthy individuals and people with MDD. In this two-center study, 62 adult patients with MDD and 71 healthy matched controls underwent high-resolution magnetic resonance imaging. We used manual tracing of the bilateral hippocampal structure with help of the software BRAINS2, assessed childhood adversity using the Childhood Trauma Questionnaire and genotyped Val66Met BDNF SNP (rs6265). MDD patients had smaller hippocampal volumes, both in the left and right hemispheres (F = 5.4, P = 0.022). We also found a significant interaction between BDNF allele and history of childhood adversity (F = 6.1, P = 0.015): Met allele carriers in our samples showed significantly smaller hippocampal volumes when they did have a history of childhood adversity, both in patients and controls. Our results highlight how relevant stress-gene interactions are for hippocampal volume reductions. Subjects exposed to early life adversity developed smaller hippocampal volumes when they carry the Met-allele of the BDNF polymorphism.

  3. The recruitment of early retirees: a vignette study of the factors that affect managers' decisions

    NARCIS (Netherlands)

    Karpinska, K.; Henkens, C.J.I.M.; Schippers, J.


    Retirement is characterised as a dynamic process that has several different outcomes, from early retirement to re-entry to the labour force. Recent studies of the Dutch population show that a substantial number of early retirees re-enter the workforce after early retirement, but others do not succee

  4. Inhibition of tyrosinase-mediated enzymatic browning by sulfite and natural alternatives

    NARCIS (Netherlands)

    Kuijpers, T.F.M.; Vincken, J.P.


    Although sulfite is widely used to counteract enzymatic browning, its mechanism has remained largely unknown. We describe a double inhibitory mechanism of sulfite on enzymatic browning, affecting both the enzymatic oxidation of phenols into o‑quinones, as well as the non‑enzymatic reactions of these


    Directory of Open Access Journals (Sweden)

    Maša Žvelc


    Full Text Available This article provides four responses from senior psychotherapists and supervisors to Erskine’s articleRelational Healing of Early Affect-Confusion. The authors approach the third part of case study trilogy from their particular perspective and provide both challenge and respect for the author’s work.

  6. The Roles of Motivation, Affective Attitudes, and Willingness to Communicate among Chinese Students in Early English Immersion Programs (United States)

    Knell, Ellen; Chi, Yanping


    Early English immersion in China has been studied from many angles, but no research to date has investigated affective variables, which may have a profound relevance to successful English acquisition. The present study examines the roles of motivation, attitudes towards learning English, willingness to communicate, perceived competence, language…

  7. Components of Negative Affect as Moderators of the Relationship between Early Drinking Onset and Binge-Drinking Behavior (United States)

    McNamara, Robert S.; Swaim, Randall C.; Rosen, Lee A.


    This study examines the moderating effects of negative affect on the relationship between early drinking onset and binge-drinking behavior. Six hundred and thirty-five eleventh- and twelfth-grade students completed the American Drug and Alcohol Survey and reported on a variety of measures, including items assessing anxiety, anger, depression, age…

  8. Structural and enzymatic insights into Lambda glutathione transferases from Populus trichocarpa, monomeric enzymes constituting an early divergent class specific to terrestrial plants. (United States)

    Lallement, Pierre-Alexandre; Meux, Edgar; Gualberto, José M; Prosper, Pascalita; Didierjean, Claude; Saul, Frederick; Haouz, Ahmed; Rouhier, Nicolas; Hecker, Arnaud


    GSTs represent a superfamily of multifunctional proteins which play crucial roles in detoxification processes and secondary metabolism. Instead of promoting the conjugation of glutathione to acceptor molecules as do most GSTs, members of the Lambda class (GSTLs) catalyse deglutathionylation reactions via a catalytic cysteine residue. Three GSTL genes (Pt-GSTL1, Pt-GSTL2 and Pt-GSTL3) are present in Populus trichocarpa, but two transcripts, differing in their 5' extremities, were identified for Pt-GSTL3. Transcripts for these genes were primarily found in flowers, fruits, petioles and buds, but not in leaves and roots, suggesting roles associated with secondary metabolism in these organs. The expression of GFP-fusion proteins in tobacco showed that Pt-GSTL1 is localized in plastids, whereas Pt-GSTL2 and Pt-GSTL3A and Pt-GSTL3B are found in both the cytoplasm and the nucleus. The resolution of Pt-GSTL1 and Pt-GSTL3 structures by X-ray crystallography indicated that, although these proteins adopt a canonical GST fold quite similar to that found in dimeric Omega GSTs, their non-plant counterparts, they are strictly monomeric. This might explain some differences in the enzymatic properties of both enzyme types. Finally, from competition experiments between aromatic substrates and a fluorescent probe, we determined that the recognition of glutathionylated substrates is favoured over non-glutathionylated forms.

  9. Photoelectrochemical enzymatic biosensors. (United States)

    Zhao, Wei-Wei; Xu, Jing-Juan; Chen, Hong-Yuan


    Enzymatic biosensors have been valuable bioanalytical devices for analysis of diverse targets in disease diagnosis, biological and biomedical research, etc. Photoelectrochemical (PEC) bioanalysis is a recently emerged method that promptly becoming a subject of new research interests due to its attractive potential for future bioanalysis with high sensitivity and specificity. PEC enzymatic biosensors integrate the inherent sensitivities of PEC bioanalysis and the selectivity of enzymes and thus share their both advantages. Currently, PEC enzymatic biosensors have become a hot topic of significant research and the recent impetus has grown rapidly as demonstrated by increased research papers. Given the pace of advances in this area, this review will make a thorough discussion and survey on the fundamentals, sensing strategies, applications and the state of the art in PEC enzymatic biosensors, followed by future prospects based on our own opinions. We hope this work could provide an accessible introduction to PEC enzymatic biosensors for any scientist.

  10. Negative Affective Spillover from Daily Events Predicts Early Response to Cognitive Therapy for Depression (United States)

    Cohen, Lawrence H.; Gunthert, Kathleen C.; Butler, Andrew C.; Parrish, Brendt P.; Wenze, Susan J.; Beck, Judith S.


    This study evaluated the predictive role of depressed outpatients' (N = 62) affective reactivity to daily stressors in their rates of improvement in cognitive therapy (CT). For 1 week before treatment, patients completed nightly electronic diaries that assessed daily stressors and negative affect (NA). The authors used multilevel modeling to…

  11. Early maternal deprivation affects dentate gyrus structure and emotional learning in adult female rats

    NARCIS (Netherlands)

    Oomen, C.A.; Soeters, H.; Audureau, N.; Vermunt, L.; van Hasselt, F.N.; Manders, E.M.M.; Joëls, M.; Krugers, H.; Lucassen, P.J.


    Rationale: Stress elicits functional and structural changes in the hippocampus. Early life stress is one of the major risk factors for stress-related pathologies like depression. Patients suffering from depression show a reduced hippocampal volume, and in women, this occurs more often when depressio

  12. Early life experiences affect the adaptive capacity of rearing hens during infectious challenges

    NARCIS (Netherlands)

    Walstra, I.; Napel, ten J.; Kemp, B.; Schipper, H.; Brand, van den H.


    This study aimed to investigate whether pre- and early postnatal experiences of rearing hens contribute to the ability to cope with infectious challenges at an older age. In a 2 × 2 factorial arrangement, 352 Lohmann Brown chicks were exposed to either suboptimal or optimized incubation plus hatch c

  13. Basic Abnormalities in Visual Processing Affect Face Processing at an Early Age in Autism Spectrum Disorder

    NARCIS (Netherlands)

    Vlamings, Petra Hendrika Johanna Maria; Jonkman, Lisa Marthe; van Daalen, Emma; van der Gaag, Rutger Jan; Kemner, Chantal


    Background: A detailed visual processing style has been noted in autism spectrum disorder (ASD); this contributes to problems in face processing and has been directly related to abnormal processing of spatial frequencies (SFs). Little is known about the early development of face processing in ASD an

  14. Bias to pollen odors is affected by early exposure and foraging experience. (United States)

    Arenas, A; Farina, W M


    In many pollinating insects, foraging preferences are adjusted on the basis of floral cues learned at the foraging site. In addition, olfactory experiences gained at early adult stages might also help them to initially choose food sources. To understand pollen search behavior of honeybees, we studied how responses elicited by pollen-based odors are biased in foraging-age workers according to (i) their genetic predisposition to collect pollen, (ii) pollen related information gained during foraging and (iii) different experiences with pollen gained at early adult ages. Bees returning to the hive carrying pollen loads, were strongly biased to unfamiliar pollen bouquets when tested in a food choice device against pure odors. Moreover, pollen foragers' orientation response was specific to the odors emitted by the pollen type they were carrying on their baskets, which suggests that foragers retrieve pollen odor information to recognize rewarding flowers outside the hive. We observed that attraction to pollen odor was mediated by the exposure to a pollen diet during the first week of life. We did not observe the same attraction in foraging-age bees early exposed to an artificial diet that did not contain pollen. Contrary to the specific response observed to cues acquired during foraging, early exposure to single-pollen diets did not bias orientation response towards a specific pollen odor in foraging-age bees (i.e. bees chose equally between the exposed and the novel monofloral pollen odors). Our results show that pollen exposure at early ages together with olfactory experiences gained in a foraging context are both relevant to bias honeybees' pollen search behavior.

  15. Early experience affects the intergenerational transmission of infant abuse in rhesus monkeys. (United States)

    Maestripieri, Dario


    Maternal abuse of offspring in macaque monkeys shares some similarities with child maltreatment in humans, including its transmission across generations. This study used a longitudinal design and a cross-fostering experiment to investigate whether abusive parenting in rhesus macaques is transmitted from mothers to daughters and whether transmission occurs through genetic or experiential factors. Nine of 16 females who were abused by their mothers in their first month of life, regardless of whether they were reared by their biological mothers or by foster mothers, exhibited abusive parenting with their firstborn offspring, whereas none of the females reared by nonabusive mothers did. These results suggest that the intergenerational transmission of infant abuse in rhesus monkeys is the result of early experience and not genetic inheritance. The extent to which the effects of early experience on the intergenerational transmission of abusive parenting are mediated by social learning or experience-induced physiological alterations remains to be established.

  16. Maternal early-life trauma and affective parenting style: the mediating role of HPA-axis function. (United States)

    Juul, Sarah H; Hendrix, Cassandra; Robinson, Brittany; Stowe, Zachary N; Newport, D Jeffrey; Brennan, Patricia A; Johnson, Katrina C


    A history of childhood trauma is associated with increased risk for psychopathology and interpersonal difficulties in adulthood and, for those who have children, impairments in parenting and increased risk of negative outcomes in offspring. Physiological and behavioral mechanisms are poorly understood. In the current study, maternal history of childhood trauma was hypothesized to predict differences in maternal affect and HPA axis functioning. Mother-infant dyads (N = 255) were assessed at 6 months postpartum. Mothers were videotaped during a 3-min naturalistic interaction, and their behavior was coded for positive, neutral, and negative affect. Maternal salivary cortisol was measured six times across the study visit, which also included an infant stressor paradigm. Results showed that childhood trauma history predicted increased neutral affect and decreased mean cortisol in the mothers and that cortisol mediated the association between trauma history and maternal affect. Maternal depression was not associated with affective measures or cortisol. Results suggest that early childhood trauma may disrupt the development of the HPA axis, which in turn impairs affective expression during mother-infant interactions in postpartum women. Interventions aimed at treating psychiatric illness in postpartum women may benefit from specific components to assess and treat trauma-related symptoms and prevent secondary effects on parenting.

  17. Does dietary protein in early life affect the development of adiposity in mammals? (United States)

    Metges, C C


    This article examines the proposition that dietary protein in pre- and early postnatal life influences the development of adiposity in later life. In rodents, low protein intake during gestation can result in low birth weight and subsequently leads to various metabolic disturbances in adulthood, such as high blood pressure, impaired glucose tolerance and insulin resistance. The few controlled studies conducted in animals suggest that high protein or energy intake during gestation leads to low birth weights. Observational studies in humans have been inconclusive in establishing a relationship between dietary protein intake in pregnancy and effects on birth weight and adiposity of the offspring later in life. There is only weak epidemiological evidence linking high protein intake during early childhood and the development of obesity. By contrast, studies in domestic animals have found that higher levels of protein intake are often associated with lower rates of fat accretion. Additional studies are proposed to explore claims linking protein nutrition in early life to the postnatal development of obesity and disease in humans.

  18. Early social environment affects the endogenous oxytocin system: a review and future directions

    Directory of Open Access Journals (Sweden)

    Emily eAlves


    Full Text Available Endogenous oxytocin plays an important role in a wide range of human functions including birth, milk ejection during lactation and facilitation of social interaction. There is increasing evidence that both variations in the oxytocin receptor (OXTR and concentrations of oxytocin are associated with differences in these functions. The causes for the differences that have been observed in tonic and stimulated oxytocin release remain unclear. Previous reviews have suggested that across the life course, these differences may be due to individual factors, e.g. genetic variation (of the OXTR, age or sex, or be the result of early environmental influences such as social experiences, stress or trauma partly by inducing epigenetic changes. This review has three aims. First, we briefly discuss the endogenous oxytocin system, including physiology, development, individual differences and function. Secondly, current models describing the relationship between the early life environment and the development of the oxytocin system in humans and animals are discussed. Finally, we describe research designs that can be used to investigate the effects of the early environment on the oxytocin system, identifying specific areas of research that need further attention.

  19. Nonsurgical scar management of the face: does early versus late intervention affect outcome? (United States)

    Parry, Ingrid; Sen, Soman; Palmieri, Tina; Greenhalgh, David


    Special emphasis is placed on the clinical management of facial scarring because of the profound physical and psychological impact of facial burns. Noninvasive methods of facial scar management include pressure therapy, silicone, massage, and facial exercises. Early implementation of these scar management techniques after a burn injury is typically accepted as standard burn rehabilitation practice, however, little data exist to support this practice. This study evaluated the timing of common noninvasive scar management interventions after facial skin grafting in children and the impact on outcome, as measured by scar assessment and need for facial reconstructive surgery. A retrospective review of 138 patients who underwent excision and grafting of the face and subsequent noninvasive scar management during a 10-year time frame was conducted. Regression analyses were used to show that earlier application of silicone was significantly related to lower Modified Vancouver Scar Scale scores, specifically in the subscales of vascularity and pigmentation. Early use of pressure therapy and implementation of facial exercises were also related to lower Modified Vancouver Scar Scale vascularity scores. No relationship was found between timing of the interventions and facial reconstructive outcome. Early use of silicone, pressure therapy, and exercise may improve scar outcome and accelerate time to scar maturity.

  20. Enzymatic desulfurization of coal

    Energy Technology Data Exchange (ETDEWEB)

    Marquis, J.K. (Boston Univ., MA (United States). School of Medicine); Kitchell, J.P. (Holometrix, Inc., Cambridge, MA (United States))


    Our current efforts to develop clean coal technology emphasize the advantages of enzymatic desulfurization techniques and have specifically addressed the potential of using partially-purified extracellular microbial enzymes or commercially available enzymes. Our work is focused on the treatment of model'' organic sulfur compounds such as dibenzothiophene (DBT) and ethylphenylsulfide (EPS). Furthermore, we are designing experiments to facilitate the enzymatic process by means of a hydrated organic solvent matrix.

  1. Parents and Early Life Environment Affect Behavioral Development of Laying Hen Chickens

    NARCIS (Netherlands)

    Haas, de E.N.; Bolhuis, J.E.; Kemp, B.; Groothuis, T.G.G.; Rodenburg, T.B.


    Severe feather pecking (SFP) in commercial laying hens is a maladaptive behavior which is associated with anxiety traits. Many experimental studies have shown that stress in the parents can affect anxiety in the offspring, but until now these effects have been neglected in addressing the problem of

  2. Happy babies, chatty toddlers: infant positive affect facilitates early expressive, but not receptive language. (United States)

    Laake, Lauren M; Bridgett, David J


    Eighty-three mother-infant dyads participated in this study. Positive affect (PA) broadly, along with fine-grained aspects of PA, was measured at 10 months of age. Language was measured at 14 months. Infant PA predicted expressive, but not receptive, language. The implications of these findings are discussed.

  3. Parents and early life environment affect behavioral development of laying hen chickens

    NARCIS (Netherlands)

    Haas, Elske N. de; Bolhuis, J. Elizabeth; Kemp, Bas; Groothuis, Ton G.G.; Rodenburg, T. Bas


    Severe feather pecking (SFP) in commercial laying hens is a maladaptive behavior which is associated with anxiety traits. Many experimental studies have shown that stress in the parents can affect anxiety in the offspring, but until now these effects have been neglected in addressing the problem of

  4. Affective facilitation of early visual cortex during rapid picture presentation at 6 and 15 Hz. (United States)

    Bekhtereva, Valeria; Müller, Matthias M


    The steady-state visual evoked potential (SSVEP), a neurophysiological marker of attentional resource allocation with its generators in early visual cortex, exhibits enhanced amplitude for emotional compared to neutral complex pictures. Emotional cue extraction for complex images is linked to the N1-EPN complex with a peak latency of ∼140-160 ms. We tested whether neural facilitation in early visual cortex with affective pictures requires emotional cue extraction of individual images, even when a stream of images of the same valence category is presented. Images were shown at either 6 Hz (167 ms, allowing for extraction) or 15 Hz (67 ms per image, causing disruption of processing by the following image). Results showed SSVEP amplitude enhancement for emotional compared to neutral images at a presentation rate of 6 Hz but no differences at 15 Hz. This was not due to featural differences between the two valence categories. Results strongly suggest that individual images need to be displayed for sufficient time allowing for emotional cue extraction to drive affective neural modulation in early visual cortex.

  5. Severity of lung fibrosis affects early surgical outcomes of lung cancer among patients with combined pulmonary fibrosis and emphysema. (United States)

    Mimae, Takahiro; Suzuki, Kenji; Tsuboi, Masahiro; Ikeda, Norihiko; Takamochi, Kazuya; Aokage, Keiju; Shimada, Yoshihisa; Miyata, Yoshihiro; Okada, Morihito


    Combined pulmonary fibrosis and emphysema (CPFE) is defined as upper lobe emphysema and lower lobe fibrosis, which are representative lung disorders that increase the prevalence of lung cancer. This unique disorder may affect the morbidity and mortality during the early period after surgery. The present study aimed to identify which clinicopathological features significantly affect early surgical outcomes after lung resection in nonsmall cell lung cancer (NSCLC) patients and in those with CPFE.We retrospectively assessed 2295 patients with NSCLC and found that 151 (6.6%) had CPFE. All were surgically treated between January 2008 and December 2010 at 4 institutions.The postoperative complication rates for patients with and without CPFE were 39% and 17%, respectively. The 90-day mortality rates were higher among patients with than without CPFE (7.9% vs 1%). Acute exacerbation of interstitial pneumonia was the main cause of death among 12 patients with CPFE who died within 90 days after surgery. Multivariate logistic regression analysis selected CPFE, gender, age, and clinical stage as independent predictive factors for postoperative complications, and CPFE, clinical stage, and sex for 90-day mortality. The severity of lung fibrosis on preoperative CT images was an independent predictive factor for 90-day mortality among patients with CPFE.The key predictive factor for postoperative mortality and complications of lung resection for NSCLC was CPFE. The severity of lung fibrosis was the principal predictor of early outcomes after lung surgery among patients with CPFE and NSCLC.

  6. Affect of surgical approaches on functional results of total hip arthroplasty in early postoperative period

    Directory of Open Access Journals (Sweden)

    D. V. Andreyev


    Full Text Available Minimally invasive approaches implies a less soft tissue damage and, therefore, more rapid recovery of the patient in the early postoperative period. The present study is a comparison of minimally invasive and standard approaches using biomechanical analysis of standing and walking patients before and after total hip arthroplasty, as well as an analysis of clinical outcomes in the early postoperative period. Fifty patients undergoing primary total hip arthroplasty using a minimally invasive and conventional techniques were divided into three groups. The first group consisted of patients operated on using the MIS AL (modified minimally invasive approach Watson-Jones (n = 17, the second - MDM (minimally invasive approach to the modified Mueller (n = 16 and in the third - with the use of transgluteal conventional approach by Harding (n = 17. The estimation of biomechanical parameters in static and dynamic patients before surgery and at 8-10 days after surgery. Also assessed clinical outcome postoperative visual analogue scale (VAS and Harris scale on day 10, 6 and 12 weeks and 1 year. When comparing the three groups of patients stabilometry best results were observed in groups of minimally invasive approaches MIS AL and MDM. When comparing the three groups significantly better (a moderate increase in the duration of the step, rolling the contralateral limb and a slight increase in the duration of the step the operated limb by increasing the duration of the roll-over were identified in the minimally invasive group MIS AL and MDM. In assessing the scale of Harris in the early postoperative period, higher rates were observed in groups of minimally invasive approaches. A year after the operation functional results become similar in all groups.

  7. Autonomic Dysfunction Predicts Early Cardiac Affection in Patients with Systemic Sclerosis

    Directory of Open Access Journals (Sweden)

    Khaled M. Othman


    Full Text Available Objective: To detect the early preclinical alterations in cardiac autonomic control as well as altered cardiac function in systemic sclerosis (SSc patients and their relevance to the clinical features of the disease using noninvasive methods. Methods: 30 SSc patients and 15 healthy controls matched for age and sex underwent clinical examination, serological analysis, and echocardiographic assessment including Doppler flow imaging to evaluate cardiac function, and 24-hour Holter monitoring analyzed for arrhythmia and heart rate variability (HRV in the time and frequency domains. Results: The trans-mitral Doppler of early to atrial wave (E/A ratio was reversed in five patients (16.6% and the tricuspid E/A ratio was reversed in 10 patients (33.3%. Holter analysis for SSc patients revealed an increased prevalence of premature ventricular contractions (PVC $ 10/h (P = 0.02, supra-ventricular tachycardias (SVTs (P = 0.2, and total PVC count (P = 0.0000. Highly significant (P = 0.000 impairment in all HRV parameters was demonstrated in the SSc patients. Total skin thickness score (TSS, Raynaud’s phenomenon and anti-scleroderma 70 (anti-SCL70 showed significant positive correlations with all arrhythmia parameters, while showing a significant negative correlation with the impaired ventricular diastolic function and various HRV parameters. No correlation was found between arrhythmia and HRV parameters and disease duration, disease type, or presence of anti-centromere antibodies. Conclusion: Low heart rate variability, increased TSS and the presence of anti-SCL70 are correlated with preclinical cardiac involvement in SSc patients and may predict the likelihood of malignant arrhythmia and sudden cardiac death. Therefore, noninvasive HRV evaluation before clinical cardiac involvement in these patients might be beneficial when added to the clinical and laboratory assessments in detecting high-risk patients, and may allow for implementation of preventive

  8. Hyaluronate degradation affects ventricular function of the early postlooped embryonic rat heart in situ. (United States)

    Baldwin, H S; Lloyd, T R; Solursh, M


    Hyaluronic acid is the major glycosaminoglycan of the early cardiac extracellular matrix or "cardiac jelly," yet little is known about its role in the ontogeny of early ventricular performance. To investigate the in situ effect of hyaluronate degradation on ventricular function, whole rat embryos were cultured in rat serum alone (control embryos) or rat serum plus 20 TRU/mL of Streptomyces hyaluronidase (treatment embryos) from gestational day 9.5 (before formation of the heart tube) through initial looping of the heart. Cardiac function was measured before looping (24 hours in culture) and immediately after looping (36 hours in culture) by video motion analysis of the external wall motion of the bulbus cordis and primitive ventricle. Degradation of hyaluronic acid in the treated embryos was confirmed by Alcian blue staining at pH 2.5. Significant increases in heart rate, circumferential shortening fraction, maximum velocity of circumferential contraction, and maximum velocity of circumferential relaxation were observed with looping in both control and treatment embryos. Although there was minimal difference in ventricular performance between control and treatment embryos before looping, there was a significant increase in all parameters of ventricular performance in the hyaluronidase-treated embryos immediately after looping of the heart. Endocardial cushions were absent in hyaluronidase-treated embryos, and an additional group of embryos cultured in the presence of Streptomyces hyaluronidase for 48 to 72 hours failed to develop endocardial cushions. These experiments are the first to (1) document a quantifiable increase in ventricular performance during early cardiac looping and (2) demonstrate that hyaluronate degradation results in abnormal endocardial cushion formation and altered ventricular performance of the postlooped heart.(ABSTRACT TRUNCATED AT 250 WORDS)

  9. Dynamic cerebral autoregulatory capacity is affected early in Type 2 diabetes

    DEFF Research Database (Denmark)

    Kim, Y.S.; Immink, R.V.; Stok, W.J.;


    ) and impaired in subjects with DM+ (30+/-5 degrees ; Pblood pressure oscillations by affected dynamic cerebral autoregulation. The steady-state response of MCA V(mean) to postural change was comparable for all groups (-12+/-6% in CTRL subjects......, -15+/-6% in subjects with DM- and -15+/-7% in subjects with DM+). HbA(1c) (glycated haemoglobin) and the duration of diabetes, but not blood pressure, were determinants of transfer function phase. In conclusion, dysfunction of dynamic cerebral autoregulation in subjects with Type 2 diabetes appears......Type 2 diabetes is associated with an increased risk of endothelial dysfunction and microvascular complications with impaired autoregulation of tissue perfusion. Both microvascular disease and cardiovascular autonomic neuropathy may affect cerebral autoregulation. In the present study, we tested...

  10. Early ERP modulation during mood adjectives processing in patients with affective disorders. (United States)

    Grzybowski, Szczepan J; Wyczesany, Miroslaw


    Attentional bias is considered a key feature in mood disorders, and yet little is known of its neural correlates within the early stream of information processing in manic and depressed patients. The study aimed to capture and detail attentional bias during emotional word processing in patients within the first 500ms of stimulus exposition. 28 mood adjectives (14 positive and 14 negative) were used as stimuli. We expected differences in adjective encoding between groups during lexico-semantic analysis based on varying attentional resource allocation. Differences were evidenced within the first 100ms after stimulus onset with higher amplitudes for both patient groups than controls, possibly indicating more automatic attention allocation to stimuli during sensory analysis stages. Between 200-290ms after the words' onset, a specific valence mood-word mismatch was registered which approached significance, with higher responses evoked to negative than positive words in manic patients, and the opposite pattern of activity observed in depressed patients, suggesting cognitive bias based on mood incongruence during the lexico-semantic analysis stage. There was also a lateralized pattern of activity, with higher amplitudes over the right hemisphere posteriorly, and higher over the left frontally in the patient groups, especially in the manic individuals. The study points to attentional bias based on mood incongruence processing during crucial stages of meaning encoding within the early stream of information processing.

  11. Specific Language Impairment affects the early spelling process quantitatively but not qualitatively. (United States)

    Cordewener, Kim A H; Bosman, Anna M T; Verhoeven, Ludo


    The present study investigated whether children with Specific Language Impairment (SLI) need a special spelling education program, by examining whether the early spelling of children with SLI is quantitatively and qualitatively different from the spelling of typically developing children. Two groups of first grade children participated: 39 children with a typical language development between the age of 73 and 88 months, and 59 children with SLI between the age of 71 and 97 months. The results indicated that children with SLI do have a quantitative delay in both grapheme knowledge and spelling during first grade. However, there was no qualitative difference between the early spelling of children with SLI and typically developing children. This indicated that children with SLI have the same spelling processes as typically developing children, although they develop slower. For clinical practice, this means that teachers of children with SLI can practice the same skills as with typically developing children, but they have to practice substantially more than typically developing children.

  12. Warming affects hatching time and early season survival of eastern tent caterpillars. (United States)

    Abarca, Mariana; Lill, John T


    Climate change is disrupting species interactions by altering the timing of phenological events such as budburst for plants and hatching for insects. We combined field observations with laboratory manipulations to investigate the consequences of climate warming on the phenology and performance of the eastern tent caterpillar (Malacosoma americanum). We evaluated the effects of warmer winter and spring regimes on caterpillar hatching patterns and starvation endurance, traits likely to be under selection in populations experiencing phenological asynchrony, using individuals from two different populations (Washington, DC, and Roswell, GA). We also quantified the proximate and extended fitness effects of early food deprivation and recorded spring phenology of local caterpillars and their host plants. In addition, we conducted laboratory assays to determine if caterpillars are using plant chemical cues to fine-tune their hatching times. Warmer winter temperatures induced earlier hatching and caterpillars from GA survived starvation for periods that were 30% longer than caterpillars from DC. Warmer spring regimes reduced the starvation endurance of caterpillars overwintering in the wild but not in the laboratory. Early starvation dramatically reduced hatchling survival; however, surviving caterpillars did not show detrimental effects on pupal mass or development time. In the field, hatching preceded budburst in both 2013 and 2014 and the period of optimal foliage quality was 2 weeks shorter in 2013. Hatching time was unaffected by exposure to plant volatiles. Overall, we found that warmer temperatures can trigger late-season asynchrony by accelerating plant phenology and caterpillars from different populations exhibit differential abilities to cope with environmental unreliability.

  13. Calcium Addition Affects Germination and Early Seedling Growth of Sweet Sorghum under Saline Conditions

    Institute of Scientific and Technical Information of China (English)


    [Objective] This study aimed to determine the interactive effects of supple- mental Ca amendment and salinity on germination of sweet sorghum seeds in saline solution culture medium, and investigate the effects of different combinations of Na/Ca ratio in saline soils on the early growth of sweet sorghum plants. [Method] A germi- nation test and a greenhouse pot experiment were conducted to assess the interac- tive effects of calcium addition to culture medium on the germination and seedling growth of sweet sorghum (Sorghum saccharatum Moench) in saline soils with a range of NaYCa ratios. In the germination test, seeds were treated with different combinations of five calcium levels [0, 5, 10, 15, and 20 mmol/L Ca(NO3)2] and five salinity levels (0, 50, 100, 150, and 200 mmol/L NaCI). In the greenhouse experi- ment, seeds were sown in potting soils containing 3 salinity levels (2.3, 4.7 and 7.0 dS/m) and three Na:Ca ratios (10:0, 10:1, and 5:1). [Result] In the germination test, Ca addition at 5 mmol/L promoted germination by 5.5%, 9.9%, and 17.0% at the 3.4, 6.7 and 10.1 dS/m salinity levels. The higher Ca level (10 mmol/L) also in- creased germination by 9.1% and 7.8% at the 3.4 and 6.7 dS/m salinity levels. Then even higher Ca addition at 15 and 20 mmol/L appeared to promote germina- tion when culture media had high salinity (10.1 and 13.4 dS/m). In the greenhouse pot experiment, saline soil amended with supplemental Ca at the 2.3 and 4.7 dS/m salinity levels significantly promoted early seedling growth, with an increase of 6.8% to 28.2% in plant height and 14.3% to 67.9% in whole plant weight. From 28 to 42 d after seeding, the relative growth of seedling was increased by Ca addition, with a reduction of 49.5% to 66.0% in plant height and 4.8% to 61.9% in whole plant weight. From 42 to 56 d after seeding, however, the relative growth of seedling was significantly inhibited by Ca amendment. [Conclusion] Results of this study indicate that appropriate supplemental

  14. Early Exercise Affects Mitochondrial Transcription Factors Expression after Cerebral Ischemia in Rats

    Directory of Open Access Journals (Sweden)

    Yongshan Hu


    Full Text Available Increasing evidence shows that exercise training is neuroprotective after stroke, but the underlying mechanisms are unknown. To clarify this critical issue, the current study investigated the effects of early treadmill exercise on the expression of mitochondrial biogenesis factors. Adult rats were subjected to ischemia induced by middle cerebral artery occlusion followed by reperfusion. Expression of two genes critical for transcriptional regulation of mitochondrial biogenesis, peroxisome proliferator-activated receptor coactivator-1 (PGC-1 and nuclear respiratory factor-1 (NRF-1, were examined by RT-PCR after five days of exercise starting at 24 h after ischemia. Mitochondrial protein cytochrome C oxidase subunit IV (COX IV was detected by Western blot. Neurological status and cerebral infarct volume were evaluated as indices of brain damage. Treadmill training increased levels of PGC-1 and NRF-1 mRNA, indicating that exercise promotes rehabilitation after ischemia via regulation of mitochondrial biogenesis.

  15. Zebrafish foxo3b negatively regulates canonical Wnt signaling to affect early embryogenesis.

    Directory of Open Access Journals (Sweden)

    Xun-wei Xie

    Full Text Available FOXO genes are involved in many aspects of development and vascular homeostasis by regulating cell apoptosis, proliferation, and the control of oxidative stress. In addition, FOXO genes have been showed to inhibit Wnt/β-catenin signaling by competing with T cell factor to bind to β-catenin. However, how important of this inhibition in vivo, particularly in embryogenesis is still unknown. To demonstrate the roles of FOXO genes in embryogenesis will help us to further understand their relevant physiological functions. Zebrafish foxo3b gene, an orthologue of mammalian FOXO3, was expressed maternally and distributed ubiquitously during early embryogenesis and later restricted to brain. After morpholino-mediated knockdown of foxo3b, the zebrafish embryos exhibited defects in axis and neuroectoderm formation, suggesting its critical role in early embryogenesis. The embryo-developmental marker gene staining at different stages, phenotype analysis and rescue assays revealed that foxo3b acted its role through negatively regulating both maternal and zygotic Wnt/β-catenin signaling. Moreover, we found that foxo3b could interact with zebrafish β-catenin1 and β-catenin2 to suppress their transactivation in vitro and in vivo, further confirming its role relevant to the inhibition of Wnt/β-catenin signaling. Taken together, we revealed that foxo3b played a very important role in embryogenesis and negatively regulated maternal and zygotic Wnt/β-catenin signaling by directly interacting with both β-catenin1 and β-catenin2. Our studies provide an in vivo model for illustrating function of FOXO transcription factors in embryogenesis.

  16. Early

    Directory of Open Access Journals (Sweden)

    Kamel Abd Elaziz Mohamed


    Conclusion: Early PDT is recommended for patients who require prolonged tracheal intubation in the ICU as outcomes like the duration of mechanical ventilation length of ICU stay and hospital stay were significantly shorter in early tracheostomy.


    Directory of Open Access Journals (Sweden)

    Richard G. Erskine


    Full Text Available This three part case study illustrates the principles, theoretical concepts, and relational methods of Integrative Psychotherapy in the treatment of a client who continually experienced early affect-confusion and lived on a “borderline” between intense neediness and rage, despair and self-reliance, impulsivity and manipulation. Part 1 describes the behavioral dynamics of a 38 year old female client who required a two-part treatment approach that emphasized an inter-subjective relationship, consistency, and respect while helping her to acknowledge and value her relational-needs and to engage in a relationally contactful form of anger.

  18. Deiodinase knockdown during early zebrafish development affects growth, development, energy metabolism, motility and phototransduction.

    Directory of Open Access Journals (Sweden)

    Enise Bagci

    Full Text Available Thyroid hormone (TH balance is essential for vertebrate development. Deiodinase type 1 (D1 and type 2 (D2 increase and deiodinase type 3 (D3 decreases local intracellular levels of T3, the most important active TH. The role of deiodinase-mediated TH effects in early vertebrate development is only partially understood. Therefore, we investigated the role of deiodinases during early development of zebrafish until 96 hours post fertilization at the level of the transcriptome (microarray, biochemistry, morphology and physiology using morpholino (MO knockdown. Knockdown of D1+D2 (D1D2MO and knockdown of D3 (D3MO both resulted in transcriptional regulation of energy metabolism and (muscle development in abdomen and tail, together with reduced growth, impaired swim bladder inflation, reduced protein content and reduced motility. The reduced growth and impaired swim bladder inflation in D1D2MO could be due to lower levels of T3 which is known to drive growth and development. The pronounced upregulation of a large number of transcripts coding for key proteins in ATP-producing pathways in D1D2MO could reflect a compensatory response to a decreased metabolic rate, also typically linked to hypothyroidism. Compared to D1D2MO, the effects were more pronounced or more frequent in D3MO, in which hyperthyroidism is expected. More specifically, increased heart rate, delayed hatching and increased carbohydrate content were observed only in D3MO. An increase of the metabolic rate, a decrease of the metabolic efficiency and a stimulation of gluconeogenesis using amino acids as substrates may have been involved in the observed reduced protein content, growth and motility in D3MO larvae. Furthermore, expression of transcripts involved in purine metabolism coupled to vision was decreased in both knockdown conditions, suggesting that both may impair vision. This study provides new insights, not only into the role of deiodinases, but also into the importance of a correct

  19. Metal cycling during sediment early diagenesis in a water reservoir affected by acid mine drainage

    DEFF Research Database (Denmark)

    Torres, Ester; Ayora, Carlos; Canovas, C. R.;


    The discharge of acid mine drainage (AMD) into a reservoir may seriously affect the water quality. To investigate the metal transfer between the water and the sediment, three cores were collected from the Sancho Reservoir (Iberian Pyrite Belt, SW Spain) during different seasons: turnover event......; oxic, stratified period; anoxic and under shallow perennially oxic conditions. The cores were sliced in an oxygen-free atmosphere, after which pore water was extracted by centrifugation and analyzed. A sequential extraction was then applied to the sediments to extract the water-soluble, monosulfide......, low crystallinity Fe(III)-oxyhydroxide, crystalline Fe(III)-oxide, organic, pyrite and residual phases. The results showed that, despite the acidic chemistry of the water column (pH

  20. Surrogate mobility and orientation affect the early neurobehavioral development of infant rhesus macaques (Macaca mulatta). (United States)

    Dettmer, Amanda M; Ruggiero, Angela M; Novak, Melinda A; Meyer, Jerrold S; Suomi, Stephen J


    A biological mother's movement appears necessary for optimal development in infant monkeys. However, nursery-reared monkeys are typically provided with inanimate surrogate mothers that move very little. The purpose of this study was to evaluate the effects of a novel, highly mobile surrogate mother on motor development, exploration, and reactions to novelty. Six infant rhesus macaques (Macaca mulatta) were reared on mobile hanging surrogates (MS) and compared to six infants reared on standard stationary rocking surrogates (RS) and to 9-15 infants reared with their biological mothers (MR) for early developmental outcome. We predicted that MS infants would develop more similarly to MR infants than RS infants. In neonatal assessments conducted at Day 30, both MS and MR infants showed more highly developed motor activity than RS infants on measures of grasping (p = .009), coordination (p = .038), spontaneous crawl (p = .009), and balance (p = .003). At 2-3 months of age, both MS and MR infants displayed higher levels of exploration in the home cage than RS infants (p = .016). In a novel situation in which only MS and RS infants were tested, MS infants spent less time near their surrogates in the first five minutes of the test session than RS infants (p = .05), indicating a higher level of comfort. Collectively, these results suggest that when nursery-rearing of infant monkeys is necessary, a mobile hanging surrogate may encourage more normative development of gross motor skills and exploratory behavior and may serve as a useful alternative to stationary or rocking surrogates.

  1. Inactivation of Tor proteins affects the dynamics of endocytic proteins in early stage of endocytosis

    Indian Academy of Sciences (India)

    Brandon Tenay; Evin Kimberlin; Michelle Williams; Juliette Denise; Joshua Fakilahyel; Kyoungtae Kim


    Tor2 is an activator of the Rom2/Rho1 pathway that regulates -factor internalization. Since the recruitment of endocytic proteins such as actin-binding proteins and the amphiphysins precedes the internalization of -factor, we hypothesized that loss of Tor function leads to an alteration in the dynamics of the endocytic proteins. We report here that endocytic proteins, Abp1 and Rvs167, are less recruited to endocytic sites not only in tor2 but also tor1 mutants. Furthermore, we found that the endocytic proteins Rvs167 and Sjl2 are completely mistargeted to the cytoplasm in tor1tor2ts double mutant cells. We also demonstrate here that the efficiency of endocytic internalization or scission in all tor mutants was drastically decreased. In agreement with the Sjl2 mislocalization, we found that in tor1tor2ts double mutant cells, as well as other tor mutant cells, the overall PIP2 level was dramatically increased. Finally, the cell wall chitin content in tor2ts and tor1tor2ts mutant cells was also significantly increased. Taken together, both functional Tor proteins, Tor1 and Tor2, are essentially required for proper endocytic protein dynamics at the early stage of endocytosis.

  2. Lipopolysaccharide exposure during the early postnatal period adversely affects the structure and function of the developing rat carotid body. (United States)

    Master, Zankhana R; Porzionato, Andrea; Kesavan, Kalpashri; Mason, Ariel; Chavez-Valdez, Raul; Shirahata, Machiko; Gauda, Estelle B


    The carotid body (CB) substantially influences breathing in premature infants by affecting the frequency of apnea and periodic breathing. In adult animals, inflammation alters the structure and chemosensitivity of the CB, yet it is not known if this pertains to neonates. We hypothesized that early postnatal inflammation leads to morphological and functional changes in the developing rat CB, which persists for 1 wk after the initial provoking insult. To test our hypothesis, we exposed rat pups at postnatal day 2 (P2) to lipopolysaccharide (LPS; 100 μg/kg) or saline (SAL) intraperitoneally. At P9-10 (1 wk after treatment), LPS-exposed animals had significantly more spontaneous intermittent hypoxic (IH) events, attenuated ventilatory responses to changes in oxygen tension (measured by whole body plethysmography), and attenuated hypoxic chemosensitivity of the carotid sinus nerve (measured in vitro), compared with SAL-exposed controls. These functional changes were associated with the following: 1) increased inflammatory cytokine mRNA levels; 2) decreased volume of supportive type II cells; and 3) elevated dopamine levels (a major inhibitory neuromodulator) within the CB. These findings suggest that early postnatal inflammation in newborn rats adversely affects the structure and function of the CB and is associated with increased frequency of intermittent desaturations, similar to the phenomenon observed in premature infants. Furthermore, this is the first newborn model of spontaneous intermittent desaturations that may be used to understand the mechanisms contributing to IH events in newborns.

  3. Does Self-Efficacy Affect Cognitive Performance in Persons with Clinically Isolated Syndrome and Early Relapsing Remitting Multiple Sclerosis?

    Directory of Open Access Journals (Sweden)

    Peter Joseph Jongen


    Full Text Available In persons with multiple sclerosis (MS a lowered self-efficacy negatively affects physical activities. Against this background we studied the relationship between self-efficacy and cognitive performance in the early stages of MS. Thirty-three patients with Clinically Isolated Syndrome (CIS and early Relapsing Remitting MS (eRRMS were assessed for self-efficacy (MSSES-18, cognition (CDR System, fatigue (MFIS-5, depressive symptoms (BDI, disease impact (MSIS-29, and disability (EDSS. Correlative analyses were performed between self-efficacy and cognitive scores, and stepwise regression analyses identified predictors of cognition and self-efficacy. Good correlations existed between total self-efficacy and Power of Attention (r= 0.65; P< 0.001, Reaction Time Variability (r= 0.57; P< 0.001, and Speed of Memory (r= 0.53; P< 0.01, and between control self-efficacy and Reaction Time Variability (r= 0.55; P< 0.01. Total self-efficacy predicted 40% of Power of Attention, 34% of Reaction Time Variability, and 40% of Speed of Memory variabilities. Disease impact predicted 65% of total self-efficacy and 58% of control self-efficacy variabilities. The findings may suggest that in persons with CIS and eRRMS self-efficacy may positively affect cognitive performance and that prevention of disease activity may preserve self-efficacy.

  4. The interaction of early life experiences with COMT val158met affects anxiety sensitivity. (United States)

    Baumann, C; Klauke, B; Weber, H; Domschke, K; Zwanzger, P; Pauli, P; Deckert, J; Reif, A


    The pathogenesis of anxiety disorders is considered to be multifactorial with a complex interaction of genetic factors and individual environmental factors. Therefore, the aim of this study was to examine gene-by-environment interactions of the genes coding for catechol-O-methyltransferase (COMT) and monoamine oxidase A (MAOA) with life events on measures related to anxiety. A sample of healthy subjects (N = 782; thereof 531 women; mean age M = 24.79, SD = 6.02) was genotyped for COMT rs4680 and MAOA-uVNTR (upstream variable number of tandem repeats), and was assessed for childhood adversities [Childhood Trauma Questionnaire (CTQ)], anxiety sensitivity [Anxiety Sensitivity Index (ASI)] and anxious apprehension [Penn State Worry Questionnaire (PSWQ)]. Main and interaction effects of genotype, environment and gender on measures related to anxiety were assessed by means of regression analyses. Association analysis showed no main gene effect on either questionnaire score. A significant interactive effect of childhood adversities and COMT genotype was observed: Homozygosity for the low-active met allele and high CTQ scores was associated with a significant increment of explained ASI variance [R(2) = 0.040, false discovery rate (FDR) corrected P = 0.04]. A borderline interactive effect with respect to MAOA-uVNTR was restricted to the male subgroup. Carriers of the low-active MAOA allele who reported more aversive experiences in childhood exhibited a trend for enhanced anxious apprehension (R(2) = 0.077, FDR corrected P = 0.10). Early aversive life experiences therefore might increase the vulnerability to anxiety disorders in the presence of homozygosity for the COMT 158met allele or low-active MAOA-uVNTR alleles.

  5. Cellular mechanisms underlying the effects of an early experience on cognitive abilities and affective states

    Directory of Open Access Journals (Sweden)

    Philippidis Eleni


    Full Text Available Abstract In the present study we investigated the effects of neonatal handling, an animal model of early experience, on spatial learning and memory, on hippocampal glucocorticoid (GR, mineralocorticoid (MR and type 1A serotonin (5-HT1A receptors, as well as brain derived neurotrophic factor (BDNF, and on circulating leptin levels, of male rats. Method Spatial learning and memory following an acute restraint stress (30 min were assessed in the Morris water maze. Hippocampal GR, MR and BDNF levels were determined immunocytochemically. 5-HT1A receptors were quantified by in vitro binding autoradiography. Circulating leptin levels, following a chronic forced swimming stress, were measured by radioimmunoassay (RIA. Data were statistically analyzed by analysis of variance (ANOVA. Results Neonatal handling increased the ability of male rats for spatial learning and memory. It also resulted in increased GR/MR ratio, BDNF and 5-HT1A receptor levels in the hippocampus. Furthermore, leptin levels, body weight and food consumption during chronic forced swimming stress were reduced as a result of handling. Conclusion Neonatal handling is shown to have a beneficial effect in the males, improving their cognitive abilities. This effect on behavior could be mediated by the handling-induced increase in hippocampal GR/MR ratio and BDNF levels. The handling-induced changes in BDNF and 5-HT1A receptors could underlie the previously documented effect of handling in preventing "depression". Furthermore, handling is shown to prevent other maladaptive states such as stress-induced hyperphagia, obesity and resistance to leptin.

  6. Gender and age interact to affect early outcome after intracerebral hemorrhage.

    Directory of Open Access Journals (Sweden)

    Odera Umeano

    Full Text Available BACKGROUND: Intracerebral hemorrhage (ICH is a common and devastating form of cerebrovascular disease. In ICH, gender differences in outcomes remain relatively understudied but have been examined in other neurological emergencies. Further, a potential effect of age and gender on outcomes after ICH has not been explored. This study was designed to test the hypothesis that age and gender interact to modify neurological outcomes after ICH. METHODS: Adult patients admitted with spontaneous primary supratentorial ICH from July 2007 through April 2010 were assessed via retrospective analysis of an existing stroke database at Duke University. Univariate analysis of collected variables was used to compare gender and outcome. Unfavorable outcome was defined as discharge to hospice or death. Using multivariate regression, the combined effect of age and gender on outcome after ICH was analyzed. RESULTS: In this study population, women were younger (61.1+14.5 versus 65.8+17.3 years, p=0.03 and more likely to have a history of substance abuse (35% versus 8.9%, p<0.0001 compared to men. Multivariable models demonstrated that advancing age had a greater effect on predicting discharge outcome in women compared to men (p=0.02. For younger patients, female sex was protective; however, at ages greater than 60 years, female sex was a risk factor for discharge to hospice or death. CONCLUSION: While independently associated with discharge to hospice or death after ICH, the interaction effect between gender and age demonstrated significantly stronger correlation with early outcome after ICH in a single center cohort. Prospective study is required to verify these findings.

  7. Atmospheric oxygen levels affect mudskipper terrestrial performance: implications for early tetrapods. (United States)

    Jew, Corey J; Wegner, Nicholas C; Yanagitsuru, Yuzo; Tresguerres, Martin; Graham, Jeffrey B


    The Japanese mudskipper (Periophthalmus modestus), an amphibious fish that possesses many respiratory and locomotive specializations for sojourns onto land, was used as a model to study how changing atmospheric oxygen concentrations during the middle and late Paleozoic Era (400-250 million years ago) may have influenced the emergence and subsequent radiation of the first tetrapods. The effects of different atmospheric oxygen concentrations (hyperoxia = 35%, normoxia = 21%, and hypoxia = 7% O2) on terrestrial performance were tested during exercise on a terrestrial treadmill and during recovery from exhaustive exercise. Endurance and elevated post-exercise oxygen consumption (EPOC; the immediate O2 debt repaid post-exercise) correlated with atmospheric oxygen concentration indicating that when additional oxygen is available P. modestus can increase oxygen utilization both during and following exercise. The time required post-exercise for mudskippers to return to a resting metabolic rate did not differ between treatments. However, in normoxia, oxygen consumption increased above hyperoxic values 13-20 h post-exercise suggesting a delayed repayment of the incurred oxygen debt. Finally, following exercise, ventilatory movements associated with buccopharyngeal aerial respiration returned to their rest-like pattern more quickly at higher concentrations of oxygen. Taken together, the results of this study show that P. modestus can exercise longer and recover quicker under higher oxygen concentrations. Similarities between P. modestus and early tetrapods suggest that increasing atmospheric oxygen levels during the middle and late Paleozoic allowed for elevated aerobic capacity and improved terrestrial performance, and likely led to an accelerated diversification and expansion of vertebrate life into the terrestrial biosphere.

  8. Early, but not late visual distractors affect movement synchronization to a temporal-spatial visual cue. (United States)

    Booth, Ashley J; Elliott, Mark T


    The ease of synchronizing movements to a rhythmic cue is dependent on the modality of the cue presentation: timing accuracy is much higher when synchronizing with discrete auditory rhythms than an equivalent visual stimulus presented through flashes. However, timing accuracy is improved if the visual cue presents spatial as well as temporal information (e.g., a dot following an oscillatory trajectory). Similarly, when synchronizing with an auditory target metronome in the presence of a second visual distracting metronome, the distraction is stronger when the visual cue contains spatial-temporal information rather than temporal only. The present study investigates individuals' ability to synchronize movements to a temporal-spatial visual cue in the presence of same-modality temporal-spatial distractors. Moreover, we investigated how increasing the number of distractor stimuli impacted on maintaining synchrony with the target cue. Participants made oscillatory vertical arm movements in time with a vertically oscillating white target dot centered on a large projection screen. The target dot was surrounded by 2, 8, or 14 distractor dots, which had an identical trajectory to the target but at a phase lead or lag of 0, 100, or 200 ms. We found participants' timing performance was only affected in the phase-lead conditions and when there were large numbers of distractors present (8 and 14). This asymmetry suggests participants still rely on salient events in the stimulus trajectory to synchronize movements. Subsequently, distractions occurring in the window of attention surrounding those events have the maximum impact on timing performance.

  9. Folate deficiency during early-mid pregnancy affects the skeletal muscle transcriptome of piglets from a reciprocal cross.

    Directory of Open Access Journals (Sweden)

    Yi Li

    Full Text Available Folate deficiency (FD during pregnancy can cause fetal intrauterine growth restriction in pigs, of which the skeletal dysplasia is a major manifestation. Factors influencing muscle development are very important in the formation of porcine meat quality trait. However, the effect of folate deficiency on skeletal muscle development and its molecular mechanisms are unknown. The objective of this study is to determine the effect of maternal folate deficiency on the skeletal muscle transcriptome of piglets from a reciprocal cross, in which full-sibling Landrace (LR and full-sibling Chinese local breed Laiwu (LW pigs were used for reciprocal cross matings, and sows were fed either a folate deficient or a normal diet during early-mid gestation. In addition, the difference in the responsiveness of the piglets to folate deficiency during early-mid pregnancy between reciprocal cross groups was investigated. Longissimus dorsi (LD muscle samples were collected from newborn piglets and a 4 × 44K Agilent porcine oligo microarray was used for transcriptome analysis of porcine LD muscle. The results showed that folate deficiency during early-mid pregnancy affected piglet body weight, LD muscle fiber number and content of intramuscular triglyceride. The microarray results indicated that 3154 genes were differentially expressed between folate deficient and normal piglets from the LR♂ × LW♀ cross, and 3885 differentially expressed genes (DEGs in the ones from the LW♂ × LR♀ cross. From functional analyses, sow folate deficiency affected almost all biological processes in the progeny. Lipid metabolism-related genes and associated metabolic pathways were regulated extensively by folate deficiency, especially in LR♂ × LW♀ cross piglets. Most of the genes that are regulated by folate deficiency in the LD muscle of piglets were different between LR♂ × LW♀ and LW♂ × LR♀ crosses, suggesting some epigenetic effects of FD exist in genes underlying

  10. Factors affecting early elementary (K--4) teachers' introduction of the nature of science: A national survey (United States)

    Sweeney, Sophia Jean

    A researcher-developed questionnaire regarding the importance and developmental appropriateness of 12 specific elements of the nature of science (Alshamrani, 2008) for early elementary (kindergarten through fourth grade [K-4]) science instruction was mailed to a random sample of U.S. K-4 teachers. At least half (N = 377) of the respondents reported introducing the inferential, empirical, creative, collaborative, tentative, and cultural aspects of the nature of science (NOS) during the current school year, as well as the idea that no single step-by-step scientific method exists. Over 90% of respondents identified the inferential, empirical, and creative aspects of the NOS as developmentally appropriate for the grade level taught. Based on a 5 point scale (0=not at all important and 4 =very important), the mean scores of eight NOS elements were above a value of three, which corresponded with the descriptor somewhat important to include in K-4 science instruction: the inferential, empirical, creative, collaborative, cultural, and tentative NOS, along with the ideas that replication is an important aspect of experimental research and that no single stepwise scientific method exists. A series of binary logistic regression analyses were used to explore the impact of three predictor variables (developmental appropriateness, importance, and presence in state standards) on teachers' self-reported introduction of each of the NOS elements during the 2009-2010 school year. The data for the presence of the NOS elements in state standards were extracted from a previous study (McComas, Lee, & Sweeney, 2009). Developmental appropriateness was a significant predictor of teachers' introduction of the NOS element for all except the collaborative, empirical and inferential aspects of NOS. Importance was a significant predictor for all 12 NOS elements of interest. A NOS element's presence in the state standards significantly predicted teachers' introduction of that NOS element for only

  11. Does lighting manipulation during incubation affect hatching rhythms and early development of sole? (United States)

    Blanco-Vives, B; Aliaga-Guerrero, M; Cañavate, J P; Muñoz-Cueto, J A; Sánchez-Vázquez, F J


    Light plays a key role in the development of biological rhythms in fish. Previous research on Senegal sole has revealed that both spawning rhythms and larval development are strongly influenced by lighting conditions. However, hatching rhythms and the effect of light during incubation are as yet unexplored. Therefore, the aim of this study was to investigate the impact of the light spectrum and photoperiod on Solea senegalensis eggs and larvae until day 7 post hatching (dph). To this end, eggs were collected immediately after spawning during the night and exposed to continuous light (LL), continuous darkness (DD), or light-dark (LD) 12L:12D cycles of white light (LD(W)), blue light (LD(B); λ(peak) = 463 nm), or red light (LD(R); λ(peak) = 685 nm). Eggs exposed to LD(B) had the highest hatching rate (94.5% ± 1.9%), whereas LD(R) and DD showed the lowest hatching rate (54.4% ± 3.9% and 48.4% ± 4.2%, respectively). Under LD conditions, the hatching rhythm peaked by the end of the dark phase, but was advanced in LD(B) (zeitgeber time 8 [ZT8]; ZT0 representing the onset of darkness) in relation to LD(W) and LD(R) (ZT11). Under DD conditions, the same rhythm persisted, although with lower amplitude, whereas under LL the hatching rhythm split into two peaks (ZT8 and ZT13). From dph 4 onwards, larvae under LD(B) showed the best growth and quickest development (advanced eye pigmentation, mouth opening, and pectoral fins), whereas larvae under LD(R) and DD had the poorest performance. These results reveal that developmental rhythms at the egg stage are tightly controlled by light characteristics, underlining the importance of reproducing their natural underwater photoenvironment (LD cycles of blue wavelengths) during incubation and early larvae development of fish.

  12. Dietary energy density affects fat mass in early adolescence and is not modified by FTO variants.

    Directory of Open Access Journals (Sweden)

    Laura Johnson

    Full Text Available BACKGROUND: Dietary energy density (DED does not have a simple linear relationship to fat mass in children, which suggests that some children are more susceptible than others to the effects of DED. Children with the FTO (rs9939609 variant that increases the risk of obesity may have a higher susceptibility to the effects of DED because their internal appetite control system is compromised. We tested the relationship between DED and fat mass in early adolescence and its interaction with FTO variants. METHODS AND FINDINGS: We carried out a prospective analysis on 2,275 children enrolled in the Avon Longitudinal Study of Parents and Children (ALSPAC. Diet was assessed at age 10 y using 3-day diet diaries. DED (kJ/g was calculated excluding drinks. Children were genotyped for the FTO (rs9939609 variant. Fat mass was estimated at age 13 y using the Lunar Prodigy Dual-energy X-ray Absorptiometry scanner. There was no evidence of interaction between DED at age 10 y and the high risk A allele of the FTO gene in relation to fat mass at age 13 y (beta = 0.005, p = 0.51, suggesting that the FTO gene has no effect on the relation between DED at 10 y and fat mass at 13 y. When DED at 10 y and the A allele of FTO were in the same model they were independently related to fat mass at 13 y. Each A allele of FTO was associated with 0.35+/-0.13 kg more fat mass at 13 y and each 1 kJ/g DED at 10 y was associated with 0.16+/-0.06 kg more fat mass at age 13 y, after controlling for misreporting of energy intake, gender, puberty, overweight status at 10 y, maternal education, TV watching, and physical activity. CONCLUSIONS: This study reveals the multi-factorial origin of obesity and indicates that although FTO may put some children at greater risk of obesity, encouraging a low dietary energy density may be an effective strategy to help all children avoid excessive fat gain.

  13. Impulsivity is associated with early sensory inhibition in neurophysiological processing of affective sounds

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    Takahiro eSoshi


    Full Text Available Impulsivity is widely related to socially problematic behaviors and psychiatric illness. Previous studies have investigated the relationship between response inhibition and impulsivity. However, no study has intensively examined how impulsivity correlates with automatic sensory processing before the drive for response inhibition to sensory inputs. Sensory gating is an automatic inhibitory function that attenuates the neural response to redundant sensory information and protects higher cognitive functions from the burst of information processing. Although sensory gating functions abnormally in several clinical populations, there is very little evidence supporting sensory gating changes in conjunction with impulsivity traits in non-clinical populations. The present study recruited healthy adults (n = 23 to conduct a neurophysiological experiment using a paired click paradigm and self-report scales assessing impulsive behavioral traits. Auditory stimuli included not only a pure tone, but also white noise, to explore the differences in auditory evoked potential responses between the two stimuli. White noise is more affective than pure tones; therefore, we predicted that the sensory gating of auditory evoked potentials (P50, N100, P200 for white noise would correlate more with self-reported impulsivity than with those for pure tones. Our main findings showed that sensory gating of the P50 and P200 amplitudes significantly correlated with self-reported reward responsiveness and fun-seeking, respectively, only for white noise stimuli, demonstrating that higher-scoring impulsivity subcomponents were related to greater sensory gating. Frequency-domain analyses also revealed that greater desynchronization of the beta band for the second white noise stimulus was associated with higher motor impulsivity scores, suggesting that an impulsivity-related change of sensory gating was associated with attentional modulation. These findings indicate that the

  14. Early, but not late visual distractors affect movement synchronization to a temporal-spatial visual cue

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    Ashley J Booth


    Full Text Available The ease of synchronising movements to a rhythmic cue is dependent on the modality of the cue presentation: timing accuracy is much higher when synchronising with discrete auditory rhythms than an equivalent visual stimulus presented through flashes. However, timing accuracy is improved if the visual cue presents spatial as well as temporal information (e.g. a dot following an oscillatory trajectory. Similarly, when synchronising with an auditory target metronome in the presence of a second visual distracting metronome, the distraction is stronger when the visual cue contains spatial-temporal information rather than temporal only. The present study investigates individuals’ ability to synchronise movements to a temporal-spatial visual cue in the presence of same-modality temporal-spatial distractors. Moreover, we investigated how increasing the number of distractor stimuli impacted on maintaining synchrony with the target cue. Participants made oscillatory vertical arm movements in time with a vertically oscillating white target dot centred on a large projection screen. The target dot was surrounded by 2, 8 or 14 distractor dots, which had an identical trajectory to the target but at a phase lead or lag of 0, 100 or 200ms. We found participants’ timing performance was only affected in the phase-lead conditions and when there were large numbers of distractors present (8 and 14. This asymmetry suggests participants still rely on salient events in the stimulus trajectory to synchronise movements. Subsequently, distractions occurring in the window of attention surrounding those events have the maximum impact on timing performance.

  15. Factors affecting early compressive strength of alkali activated fly ash (OPC-free concrete

    Directory of Open Access Journals (Sweden)

    Palomo, A.


    Full Text Available This paper presents the findings of experimental research into the chief characteristics of a new type of concrete made solely with alkali activated fly ash (AAFA: i.e., free of ordinary Portland cement (OPC. The results of testing to determine specific properties of the fresh concrete and the development of its mechanical strength showed that most of the factors that affect the manufacture and final properties of Portland cement concrete (water/cement ratio, curing conditions, etc. also impact the preparation and final quality of this new material. A number of parameters specific to AAFA concrete (nature and concentration of alkali present in the system were also explored to determine their role in the setting and hardening process.Este trabajo presenta los resultados de una investigación experimental llevada a cabo para evaluar las principales características de un nuevo tipo de hormigón fabricado solamente con ceniza volante activada alcalinamente (AAFA; es decir, sin cemento Portland comercial (OPC. Los resultados de los ensayos realizados para determinar las propiedades específicas del hormigón fresco y el desarrollo de resistencias mecánicas mostraron que la mayoría de los factores que afectan al proceso de fabricación y a las propiedades finales de los hormigones de cemento Portland (relación agua/cemento, condiciones de curado, etc. también afectan a la preparación y calidad final de estos nuevos materiales. También fueron estudiados otros parámetros específicos de los hormigones de AAFA (la naturaleza y concentración del álcali presente en el sistema para determinar su papel en el proceso de fraguado y endurecimiento.

  16. Neonatal stress-induced affective changes in adolescent Wistar rats: early signs of schizophrenia-like behavior

    Directory of Open Access Journals (Sweden)

    Carlos Eduardo Neves Girardi


    Full Text Available Psychiatric disorders are multifactorial diseases with etiology that may involve genetic factors, early life environment and stressful life events. The neurodevelopmental hypothesis of schizophrenia is based on a wealth of data on increased vulnerability in individuals exposed to insults during the perinatal period. Maternal deprivation disinhibits the adrenocortical response to stress in neonatal rats and has been used as an animal model of schizophrenia. To test if long-term affective consequences of early life stress were influenced by maternal presence, we submitted 10-day old rats, either deprived (for 22 h or not from their dams, to a stress challenge (i.p. saline injection. Corticosterone plasma levels were measured 2 h after the challenge, whereas another subgroup was assessed for behavior in the open field, elevated plus maze, social investigation and the negative contrast sucrose consumption test in adolescence (postnatal day 45. Maternally deprived rats exhibited increased plasma corticosterone levels which were higher in maternally deprived and stress challenged pups. Social investigation was impaired in maternally deprived rats only, while saline injection, independently of maternal deprivation, was associated with increased anxiety-like behavior in the elevated plus maze and an impaired intake decrement in the negative sucrose contrast. In the open field, center exploration was reduced in all maternally-deprived adolescents and in control rats challenged with saline injection. The most striking finding was that exposure to a stressful stimulus per se, regardless of maternal deprivation, was linked to differential emotional consequences. We therefore propose that besides being a well-known and validated model of schizophrenia in adult rats, the maternal deprivation paradigm could be extended to model early signs of psychiatric dysfunction, and would particularly be a useful tool to detect early signs that resemble schizophrenia.

  17. Neonatal stress-induced affective changes in adolescent Wistar rats: early signs of schizophrenia-like behavior (United States)

    Girardi, Carlos Eduardo Neves; Zanta, Natália Cristina; Suchecki, Deborah


    Psychiatric disorders are multifactorial diseases with etiology that may involve genetic factors, early life environment and stressful life events. The neurodevelopmental hypothesis of schizophrenia is based on a wealth of data on increased vulnerability in individuals exposed to insults during the perinatal period. Maternal deprivation (MD) disinhibits the adrenocortical response to stress in neonatal rats and has been used as an animal model of schizophrenia. To test if long-term affective consequences of early life stress were influenced by maternal presence, we submitted 10-day old rats, either deprived (for 22 h) or not from their dams, to a stress challenge (i.p. saline injection). Corticosterone plasma levels were measured 2 h after the challenge, whereas another subgroup was assessed for behavior in the open field, elevated plus maze (EPM), social investigation and the negative contrast sucrose consumption test in adolescence (postnatal day 45). Maternally deprived rats exhibited increased plasma corticosterone (CORT) levels which were higher in maternally deprived and stress challenged pups. Social investigation was impaired in maternally deprived rats only, while saline injection, independently of MD, was associated with increased anxiety-like behavior in the EPM and an impaired intake decrement in the negative sucrose contrast. In the open field, center exploration was reduced in all maternally-deprived adolescents and in control rats challenged with saline injection. The most striking finding was that exposure to a stressful stimulus per se, regardless of MD, was linked to differential emotional consequences. We therefore propose that besides being a well-known and validated model of schizophrenia in adult rats, the MD paradigm could be extended to model early signs of psychiatric dysfunction, and would particularly be a useful tool to detect early signs that resemble schizophrenia. PMID:25309370

  18. Early application of nerve growth factor affects serum inflammatory cytokine levels in neonatal hypoxic ischemic encephalopathy

    Institute of Scientific and Technical Information of China (English)


    BACKGROUND: It has been demonstrated that there are changes of various cytokines, chemokines and adhesion factors in neonatal hypoxic ischemic encephalopathy (HIE). What are the changes of interleukin-6 and interleukin-18 in serum of HIE neonates.OBJECTIVE: To observe the dynamic changes of interleukin-6 and interleukin-18 in peripheral serum at different time after HIE in neonates, and analyze the possible therapeutic efficacy of early application of NGF.DESIGN: A non-randomized controlled observation synchronically.SETTING: Department of Neonatology, Sun Yat-sen Hospital affiliated to Sun Yat-sen University.PARTICIPANTS: Sixty neonates with HIE were selected from the Department of Neonatology, Sun Yat-sen Hospital affiliated to Sun Yat-sen University from January 2004 to October 2006, including 32 boys and 28 girls, who were all accorded with the diagnostic standards for moderate to severe HIE. The neonates were divided into two groups NGF-treated group (n =30), HIE group (n =30). The HIE neonates in the NGF-treated group were given routine treatment and intramuscular injection of NGF within 24 hours after birth. Those in the HIE group were given routine treatments. Meanwhile, 30 apneic normal neonates (17 boys and 13 girls) at the same period were selected as the control group. The gestational age was 37 - 42 weeks in all the three groups, the body mass at birth was 2 500 - 4 000 g. Informed contents were obtained from the relatives of all the enrolled neonates.METHODS: The HIE neonates in the NGF-treated group were given routine treatment and intramuscular injection of NGF (2 000 U) within 24 hours after birth, once a day, 10 days as a course. Those in the HIE group were given routine treatments. Blood samples (3 mL) were drawn from femoral vein in all the neonates 1, 3 and 7 days after birth. The levels of interleukin-6 and interleukin-18 in serum were detected with enzyme-linked immunoabsorbent assay (ELISA).MAIN OUTCOME MEASURES: Serum levels of interlenkin

  19. A longitudinal investigation of the affective environment in families with young children: from infancy to early school age. (United States)

    Barry, Robin A; Kochanska, Grazyna


    We examined the affective environment in 102 families studied longitudinally when children were 7, 15, 25, 38, 52, and 67 months of age. At each assessment, every mother-child and father-child dyad was observed in typical daily contexts. Each person's emotions of affection, joy, and anger were coded. Both parents rated marital satisfaction when children were 15, 52, and 67 months. Growth curve analyses, using Actor-Partner Interdependence Modeling, examined (a) developmental changes in emotions, (b) within-relationship influences of the partner's emotions, (c) across-relationship influences of emotions in other parent's interactions with the child, and (d) associations between marital quality and emotions over time. Parents' emotional expressiveness was highest early in the child's development, and declined thereafter. Children's anger was highest at 15 months of age, and declined thereafter, and their positive emotions, particularly with mothers, increased over time. Generally, one's positive emotions and better marital quality were associated with greater positive emotion within- and across-relationships, whereas one's anger was associated with greater anger within- and across-relationships. However, any emotion expression elicited greater affection in the interaction partner. Parents' neuroticism did not account for the convergence of emotions across relationships.



    Reyna M., L.; Facultad de Química e ingeniería Química, Universidad Nacional Mayor de San Marcos. Lima-Perú; Robles, R.; Facultad de Química e ingeniería Química, Universidad Nacional Mayor de San Marcos. Lima-Perú; Reyes P., M.; Facultad de Química e ingeniería Química, Universidad Nacional Mayor de San Marcos. Lima-Perú; Mendoza R., Y.; Facultad de Química e ingeniería Química, Universidad Nacional Mayor de San Marcos. Lima-Perú; Romero D., J.; Facultad de Química e ingeniería Química, Universidad Nacional Mayor de San Marcos. Lima-Perú


    We obtained rate constant for enzymatic hydrolysis of barley at temperature of 70 ºC. Se ha llevado a cabo la hidrólisis enzimática de la cebada a 70 ºC y se determinó la constante cinética de reacción.

  1. Enzymatic synthesis of vanillin

    NARCIS (Netherlands)

    van den Heuvel, RHH; Fraaije, MW; Laane, C; van Berkel, WJH; Heuvel, Robert H.H. van den; Berkel, Willem J.H. van


    Due to increasing interest in natural vanillin, two enzymatic routes for the synthesis of vanillin were developed. The flavoprotein vanillyl alcohol oxidase (VAO) acts on a wide range of phenolic compounds and converts both creosol and vanillylamine to vanillin with high yield. The VAO-mediated conv

  2. Inhibitory repetitive transcranial magnetic stimulation (rTMS) of the dorsolateral prefrontal cortex modulates early affective processing. (United States)

    Zwanzger, Peter; Steinberg, Christian; Rehbein, Maimu Alissa; Bröckelmann, Ann-Kathrin; Dobel, Christian; Zavorotnyy, Maxim; Domschke, Katharina; Junghöfer, Markus


    The dorsolateral prefrontal cortex (dlPFC) has often been suggested as a key modulator of emotional stimulus appraisal and regulation. Therefore, in clinical trials, it is one of the most frequently targeted regions for non-invasive brain stimulation such as repetitive transcranial magnetic stimulation (rTMS). In spite of various encouraging reports that demonstrate beneficial effects of rTMS in anxiety disorders, psychophysiological studies exploring the underlying neural mechanisms are sparse. Here we investigated how inhibitory rTMS influences early affective processing when applied over the right dlPFC. Before and after rTMS or sham stimulation, subjects viewed faces with fearful or neutral expressions while whole-head magnetoencephalography (MEG) was recorded. Due to the disrupted functioning of the right dlPFC, visual processing in bilateral parietal, temporal, and occipital areas was amplified starting at around 90 ms after stimulus onset. Moreover, increased fear-specific activation was found in the right TPJ area in a time-interval between 110 and 170 ms. These neurophysiological effects were reflected in slowed reaction times for fearful, but not for neutral faces in a facial expression identification task while there was no such effect on a gender discrimination control task. Our study confirms the specific and important role of the dlPFC in regulation of early emotional attention and encourages future clinical research to use minimal invasive methods such as transcranial magnetic (TMS) or direct current stimulation (tDCS).

  3. High levels of inorganic nutrients affect fertilization kinetics, early development and settlement of the scleractinian coral Platygyra acuta (United States)

    Lam, E. K. Y.; Chui, A. P. Y.; Kwok, C. K.; Ip, A. H. P.; Chan, S. W.; Leung, H. N.; Yeung, L. C.; Ang, P. O.


    Dose-response experiments were conducted to investigate the effects of ammonia nitrogen (NH3/NH4 +) and orthophosphate (PO4 3-) on four stages of larval development in Platygyra acuta, including fertilization, embryonic development and the survival, motility, and settlement of planula larvae. Fertilization success was reduced significantly under 200 μM NH3/NH4 + or PO4 3-. These high doses of NH3/NH4 + and PO4 - affected egg viability (or sperm viability and polyspermic block simultaneously) and polyspermic block, respectively. These results provide the first evidence to indicate the mechanisms of how inorganic nutrients might affect coral fertilization processes. For embryonic development, NH3/NH4 + at 25-200 μM caused delay in cell division after 2-h exposure and NH3/NH4 + at 100-200 μM resulted in larval death after 72 h. However, no significant differences were observed in the mobility and survivorship of either planula or competent larvae under different levels of NH3/NH4 + or PO4 3-. There was a significant (~30 %) drop in the settlement of competent larvae under the combined effect of 100 μM NH3/NH4 + and PO4 3-. The effects of elevated nutrients appeared to become more significant only on gametes or larvae undergoing active cellular activities at fertilization, early development, and settlement.

  4. Enzymatic production of cyclodextrins. (United States)

    Biwer, A; Antranikian, G; Heinzle, E


    Cyclodextrins (CD) are enzymatically modified starches with a wide range of applications in food, pharmaceutical and chemical industries, agriculture and environmental engineering. They are produced from starch via enzymatic conversion using cyclodextrin glycosyl transferases (CGTases) and partly alpha-amylases. Due to its low solubility in water, separation and purification of beta-CD is relatively easy compared to alpha- and gamma-CD. In recent years more economic processes for gamma-CD and especially alpha-CD production have been developed using improved CGTases and downstream processing. New purification steps, e.g. affinity adsorption, may reduce the use of complexing agents. The implementation of thermostable CGTases can simplify the production process and increase the selectivity of the reaction. A tabular overview of alpha-CD production processes is presented.

  5. Enzymatic desulfurization of coal

    Energy Technology Data Exchange (ETDEWEB)

    Boyer, Y.N.; Crooker, S.C.; Kitchell, J.P.; Nochur, S.V.


    The overall objective of this program was to investigate the feasibility of an enzymatic desulfurization process specifically intended for organic sulfur removal from coal. Toward that end, a series of specific objectives were defined: (1) establish the feasibility of (bio)oxidative pretreatment followed by biochemical sulfate cleavage for representative sulfur-containing model compounds and coals using commercially-available enzymes; (2) investigate the potential for the isolation and selective use of enzyme preparations from coal-utilizing microbial systems for desulfurization of sulfur-containing model compounds and coals; and (3) develop a conceptual design and economic analysis of a process for enzymatic removal of organic sulfur from coal. Within the scope of this program, it was proposed to carry out a portion of each of these efforts concurrently. (VC)

  6. Tacrolimus does not affect early wound healing in a rodent model of bowel anastomoses and abdominal wall closure.

    Directory of Open Access Journals (Sweden)

    Martine C M Willems

    Full Text Available BACKGROUND: Use of immunosuppressant drugs has been associated with complications in wound healing. The calcineurin inhibitor tacrolimus is thought to have a relatively low complication rate, but preclinical research has yielded contradictory data, prompting the current comprehensive study. METHODS: Three groups of 33 male Wistar rats received a daily subcutaneous dose of 0,5, 2 or 5 mg/kg tacrolimus. A control group received saline. On day 0 a resection of 1 cm ileum and 1 cm colon was performed, and end-to-end anastomoses were constructed. Ten rats of each group were killed on day 3 and day 5 and the remaining animals on day 7. Both anastomoses and the wound in the abdominal wall were analyzed. Wound strength was the primary outcome parameter. RESULTS: Mean strength of the abdominal wall increased significantly over time in all groups (p<0.0001. Both the breaking strength and the bursting pressure of the ileum and colon anastomoses followed the same pattern. No differences were observed between control and experimental groups. In addition, no consistent differences were found between groups regarding wound hydroxyproline content and the activities of matrix metalloproteinase-2 and -9. CONCLUSION: Tacrolimus does not affect early wound healing.


    Directory of Open Access Journals (Sweden)

    Preeti Tabitha LOUIS


    Full Text Available The present study adopts a randomized experimental design to evaluate the impact of a father-mediated therapy to improve the play skills, affect, language, social skills and behavior among 30 clinically diagnosed autistic children at the age of 3-5 years. Standardized inventories such as, The Play Based Observation (PBO, The Griffiths Mental Developmental Scales (GMDS, The Vineland Social Maturity Scale (VSMS and the Rendel Shorts Questionnaire were administered pre and post intervention. A special program that involved fathers in the caregiving and nurturing processes of these children was designed and implemented for 6 months after which the children were reassessed. Prior to the intervention, deficits in play skills and developmental delays across expressive and receptive language were observed Scores on the Vineland Social Maturity Scale and the Rendel Shorts revealed behavioral markers. Post intervention, we noticed significant differences in the play, language acquisition, social engagement and behavior in the treatment group in comparison to the control group. The results suggested that father-mediated therapeutic involvement significantly has proven to positively foster development in young autistic children and this is an important implication for practitioners in developing early intervention programs.

  8. Enzymatic cascade bioreactor (United States)

    Simmons, Blake A.; Volponi, Joanne V.; Ingersoll, David; Walker, Andrew


    Disclosed is an apparatus and method for continuously converting sucrose to .beta.-D-glucose. The method comprises a three stage enzymatic reactor in which an aqueous solution of sucrose is first converted into a solution of fructose and .alpha.-D-glucose by passing it through a porous, packed column containing an inert media on which invertase is immobilized. This solution is then sent through a second packed column containing glucose isomerase and finally a third packed column containing mutarotase. Solution temperature and pH are adjusted to maximize glucose output.

  9. Enzymatic Modification of Sphingomyelin

    DEFF Research Database (Denmark)

    Zhang, Long; Hellgren, Lars; Xu, Xuebing

    Due to its major role in maintaining the water-retaining properties of the epidermis, ceramide is of great commercial potential in cosmetic and pharmaceuticals such as hair and skin care products. Currently, chemical synthesis of ceramide is a costly process, and developments of alternative cost......-efficient, high yield production methods are of great interest. In the present study, the potential of producing ceramide through enzymatic hydrolysis of sphingomyelin have been studied. Sphingomyelin (SM) is a ubiquitous membrane-lipid and dairy products or by-products is a rich source of sphingomyelin....... In present study, we have optimized the production of ceramide from sphingomyelin using Phospholipase C from Clostridium perfringens....

  10. Does Early Childhood Teacher Education Affect Students' Cognitive Orientations? The Effect of Different Education Tracks in Teacher Education on Prospective Early Childhood Teachers' Cognitive Orientations in Germany (United States)

    Mischo, Christoph; Wahl, Stefan; Strohmer, Janina; Wolf, Carina


    Early childhood teachers may differ regarding the knowledge base they use when making professional decisions. In this study two orientations are distinguished: the orientation towards scientific knowledge vs. the orientation towards intuition and subjective experience. As different tracks in early childhood teacher education qualify for…

  11. Changes in enzymatic activities and microbial properties in vermicompost of water hyacinth as affected by pre-composting and fungal inoculation: a comparative study of ergosterol and chitin for estimating fungal biomass. (United States)

    Pramanik, P


    In this experiment, three different fungal species, viz. Trichoderma viridae, Aspergillus niger and Phanerochaete chrysosporium, were inoculated in 7 day and 15 day partially decomposed water hyacinth to study their effect on enzymatic activities, microbial respiration and fungal biomass of the final stabilized product. The results suggested that increasing the duration of pre-composting from 7 days to 15 days did not show any significant effect on the activities of hydrolytic enzymes. Inoculation of fungi significantly (P vermicomposts. Inoculation of P. chrysosporium in initial organic waste registered the highest chitin content in vermicompost. A comparison of fungal biomass and chitin content revealed a conversion factor of 2.628 with a standard deviation of 0.318. Due to significant correlation (r = 0.864), this conversion factor allows for the calculation of fungal biomass from chitin, which is comparatively more stable than ergosterol.

  12. 影响荔枝果肉酶促褐变主要酶性质的比较研究%Comparative Research on Chracterization of Main Enzymes Affecting Enzymatic Browning of Litchi Pulp

    Institute of Scientific and Technical Information of China (English)

    刘春丽; 陈欲云


    This thesis did a comparative study on chracterization of Polyphenolxidase and Peroxidaseduring the enzymatic browning of litchi pulp. The activity of POD is far greater than the activity of PPO,however, the thermal stability of PPO was higher than that of PPO, therefore the polyphenol oxidase was adopted as the indicator of inactivating enzyme. And the activity of peroxidase can be completely inhibited by the glutathione , but the activity of polyphenol oxidase e can be inhibited transiently by the glutathione.The polyphenol oxidase and peroxidase of the pH stability was the same, at the pH less than 5 that under acidic conditions, two kinds of enzyme activity and stability dropped, thereby inhibiting the enzymatic browning.%比较研究了在荔枝果肉酶促褐变过程中,多酚氧化酶和过氧化物两种酶的性质。在荔枝果肉中过氧化物酶的活性远远大于多酚氧化酶的活性。但是PPO的热稳定性高于POD的热稳定性,应以多酚氧化酶作为灭酶的指标;并且有化学抑制剂可以完全抑制过氧化物酶活性,但对多酚氧化酶却具有抑制暂时性。pH对多酚氧化酶和过氧化物酶的稳定性影响基本一致,在pH小于5的酸性条件下活性和稳定性下降,因而在酸性条件下,可有效抑制酶促褐变。

  13. Enzymatic Hydrolysis of Lignocelluloses

    DEFF Research Database (Denmark)

    Kolasa, Marta; Ahring, Birgitte Kiær; Lübeck, Peter Stephensen;


    Lignocellulosic materials form a huge part of the plant biomass from agricultural and forestry wastes. They consist of three major components: cellulose, hemicellulose and lignin. Cellulose, the main constituent of plant cell wall, is a polymer of D–glucopyranose units linked by β-1,4-glucosidic...... bonds. Cellulose can be degraded to simple sugar components by means of enzymatic hydrolysis. However, due to its complex, crystalline structure it is difficult to break it down and the cooperative action of a variety of cellulolytic enzymes is necessary. Fungi are known to have potential in production...... using suppression subtractive hybridization and a targeted genomic screening approach using degenerate PCR. Suppression subtractive hybridization facilitates identification of genes encoding cellulolytic enzymes that are expressed when cultivating a fungal strain in medium with cellulose as the carbon...

  14. Enzymatic Modification of Sphingomyelin

    DEFF Research Database (Denmark)

    Due to its major role in maintaining the water-retaining properties of the epidermis, ceramide is of great commercial potential in cosmetic and pharmaceuticals such as hair and skin care products. Currently, chemical synthesis of ceramide is a costly process, and developments of alternative cost......-efficient, high yield production methods are of great interest. In the present study, the potential of producing ceramide through the enzymatic hydrolysis of sphingomyelin have been studied. sphingomyelin is a ubiquitous membrane-lipid and rich in dairy products or by-products. It has been verified...... that sphingomyelin modification gives a feasible approach to the potential production of ceramide. The reaction system has been improved through system evaluation and the optimization of several important factors, and phospholipase C from Clostridium perfringens shows higher activity towards the hydrolysis reaction...

  15. Pulmonary exposure to single-walled carbon nanotubes does not affect the early immune response against Toxoplasma gondii

    Directory of Open Access Journals (Sweden)

    Swedin Linda


    Full Text Available Abstract Background Single-walled carbon nanotubes (SWCNT trigger pronounced inflammation and fibrosis in the lungs of mice following administration via pharyngeal aspiration or inhalation. Human exposure to SWCNT in an occupational setting may occur in conjunction with infections and this could yield enhanced or suppressed responses to the offending agent. Here, we studied whether the sequential exposure to SWCNT via pharyngeal aspiration and infection of mice with the ubiquitous intracellular parasite Toxoplasma gondii would impact on the immune response of the host against the parasite. Methods C57BL/6 mice were pre-exposed by pharyngeal administration of SWCNT (80 + 80 μg/mouse for two consecutive days followed by intravenous injection with either 1x103 or 1x104 green fluorescence protein and luciferase-expressing T. gondii tachyzoites. The dissemination of T. gondii was monitored by in vivo bioluminescence imaging in real time for 7 days and by plaque formation. The inflammatory response was analysed in bronchoalveolar lavage (BAL fluid, and by assessment of morphological changes and immune responses in lung and spleen. Results There were no differences in parasite distribution between mice only inoculated with T. gondii or those mice pre-exposed for 2 days to SWCNT before parasite inoculum. Lung and spleen histology and inflammation markers in BAL fluid reflected the effects of SWCNT exposure and T. gondii injection, respectively. We also noted that CD11c positive dendritic cells but not F4/80 positive macrophages retained SWCNT in the lungs 9 days after pharyngeal aspiration. However, co-localization of T. gondii with CD11c or F4/80 positive cells could not be observed in lungs or spleen. Pre-exposure to SWCNT did not affect the splenocyte response to T. gondii. Conclusions Taken together, our data indicate that pre-exposure to SWCNT does not enhance or suppress the early immune response to T. gondii in mice.

  16. Early life exposure to artificial light at night affects the physiological condition: An experimental study on the ecophysiology of free-living nestling songbirds. (United States)

    Raap, Thomas; Casasole, Giulia; Pinxten, Rianne; Eens, Marcel


    Light pollution or artificial light at night (ALAN) is increasingly recognised to be an important anthropogenic environmental pressure on wildlife, affecting animal behaviour and physiology. Early life experiences are extremely important for the development, physiological status and health of organisms, and as such, early exposure to artificial light may have detrimental consequences for organism fitness. We experimentally manipulated the light environment of free-living great tit nestlings (Parus major), an important model species in evolutionary and environmental research. Haptoglobin (Hp) and nitric oxide (NOx), as important indicators of immunity, health, and physiological condition, were quantified in nestlings at baseline (13 days after hatching) and after a two night exposure to ALAN. We found that ALAN increased Hp and decreased NOx. ALAN may increase stress and oxidative stress and reduce melatonin which could subsequently lead to increased Hp and decreased NOx. Haptoglobin is part of the immune response and mounting an immune response is costly in energy and resources and, trade-offs are likely to occur with other energetically demanding tasks, such as survival or reproduction. Acute inhibition of NOx may have a cascading effect as it also affects other physiological aspects and may negatively affect immunocompetence. The consequences of the observed effects on Hp and NOx remain to be examined. Our study provides experimental field evidence that ALAN affects nestlings' physiology during development and early life exposure to ALAN could therefore have long lasting effects throughout adulthood.

  17. Combined enzymatic starch hydrolysis

    Energy Technology Data Exchange (ETDEWEB)

    Nebesny, E.


    From researches so far there comes out that glucoamylase AMG 300 L and pullulanase Promozyme 200 L when used in quantities the same as in preparation of Dextrozyme 225/75 L Novo at an action on liquified starch by means of /alpha/-amylase after 48 h of saccharification already (similarly like Dextrozyme) are able to get up to 98 DE. Chromatographic analysis proved that glucoamylase AMG 300 L Novo and succouring it pullulanase Promozyme 200 L are working most effectively when both enzymes are added to the liquified starch medium simultaneously. From this comes out that pullulanase hydrolyzes better /alpha/-1,6 bonds in lowmolecular dextrins than in oligosaccharides G/sub 4/ to G/sub 7/ formed at previous action of glucoamylase. At an optimum ratio of glucoamylase and pullulanase in relation to the dissolved starch after 8 h of the hydrolysis there are neither iso-sugars (isomaltose, panose), no oligosaccharides higher than G/sub 5/ and no dextrins. At the solution of the starch by /alpha/-amylase and its hydrolysis by enzymatic preparation Fungamyl 800 L Novo, at doses 0.02-0.8% to d.s. of starch, already after 8 h the reaction of hydrolysis contents of 36-62% maltose in dry substance of hydrolyzates are reached with only traces of glucose. (orig.).

  18. Enzymatic degradation of endomorphins. (United States)

    Janecka, Anna; Staniszewska, Renata; Gach, Katarzyna; Fichna, Jakub


    Centrally acting plant opiates, such as morphine, are the most frequently used analgesics for the relief of severe pain, even though their undesired side effects are serious limitation to their usefulness. The search for new therapeutics that could replace morphine has been mainly focused on the development of peptide analogs or peptidomimetics with high selectivity for one receptor type and high bioavailability, that is good blood-brain barrier permeability and enzymatic stability. Drugs, in order to be effective, must be able to reach the target tissue and to remain metabolically stable to produce the desired effects. The study of naturally occurring peptides provides a rational and powerful approach in the design of peptide therapeutics. Endogenous opioid peptides, endomorphin-1 and endomorphin-2, are two potent and highly selective mu-opioid receptor agonists, discovered only a decade ago, which display potent analgesic activity. However, extensive studies on the possible use of endomorphins as analgesics instead of morphine met with failure due to their instability. This review deals with the recent investigations that allowed determine degradation pathways of endomorphins in vitro and in vivo and propose modifications that will lead to more stable analogs.

  19. Molecular crowding and protein enzymatic dynamics. (United States)

    Echeverria, Carlos; Kapral, Raymond


    The effects of molecular crowding on the enzymatic conformational dynamics and transport properties of adenylate kinase are investigated. This tridomain protein undergoes large scale hinge motions in the course of its enzymatic cycle and serves as prototype for the study of crowding effects on the cyclic conformational dynamics of proteins. The study is carried out at a mesoscopic level where both the protein and the solvent in which it is dissolved are treated in a coarse grained fashion. The amino acid residues in the protein are represented by a network of beads and the solvent dynamics is described by multiparticle collision dynamics that includes effects due to hydrodynamic interactions. The system is crowded by a stationary random array of hard spherical objects. Protein enzymatic dynamics is investigated as a function of the obstacle volume fraction and size. In addition, for comparison, results are presented for a modification of the dynamics that suppresses hydrodynamic interactions. Consistent with expectations, simulations of the dynamics show that the protein prefers a closed conformation for high volume fractions. This effect becomes more pronounced as the obstacle radius decreases for a given volume fraction since the average void size in the obstacle array is smaller for smaller radii. At high volume fractions for small obstacle radii, the average enzymatic cycle time and characteristic times of internal conformational motions of the protein deviate substantially from their values in solution or in systems with small density of obstacles. The transport properties of the protein are strongly affected by molecular crowding. Diffusive motion adopts a subdiffusive character and the effective diffusion coefficients can change by more than an order of magnitude. The orientational relaxation time of the protein is also significantly altered by crowding.

  20. Homogeneous, Heterogeneous, and Enzymatic Catalysis. (United States)

    Oyama, S. Ted; Somorjai, Gabor A.


    Discusses three areas of catalysis: homegeneous, heterogeneous, and enzymatic. Explains fundamentals and economic impact of catalysis. Lists and discusses common industrial catalysts. Provides a list of 107 references. (MVL)

  1. Effects of thermal and enzymatic treatments and harvesting time on the microbial quality and chemical composition of fibre hemp (Cannabis sativa L.)

    Energy Technology Data Exchange (ETDEWEB)

    Nykter, Minna; Kymaelaeinen, Hanna-Riitta; Sjoeberg, Anna-Maija [Department of Agrotechnology, University of Helsinki, P.O. Box 28, FI-00014 University of Helsinki (Finland); Thomsen, Anne Belinda; Thygesen, Anders [Biosystems Department, Risoe National Laboratory, P.O. Box 49, DK-4000 Roskilde (Denmark); Lilholt, Hans [Materials Research Department, Risoe National Laboratory, P.O. Box 49, DK-4000 Roskilde (Denmark); Koponen, Hilkka [Department of Applied Biology, Section of Plant Pathology, University of Helsinki, P.O. Box 27, FI-00014 University of Helsinki (Finland)


    The aim of the present study was to examine the effects of pectinase enzyme treatment followed by thermal treatments (steam explosion and dry heating) on the microbial quality and chemical composition of hemp fibres. Before these treatments, the fibres were separated manually from the stems harvested after stand retting in the field before frost, after early frost or in the following spring. The enzymatic treatment of hemp promoted growth of moulds on the fibres (500-fold increase in colony-forming units (cfu)), whereas steam explosion reduced the amount of moulds to a relatively constant level of 10{sup 2} cfu/g dw. The amount of bacteria was not markedly affected by enzymatic treatment but was reduced tenfold after steam explosion. Steam explosion is thereby a potentially good process for the production of hemp fibres with low fungal contamination, which can be of importance in insulation materials. Dry heating had no effect on mould and bacterial counts at temperatures below 120 C and durations less than 60 min. The chemical composition was affected by the enzymatic treatment due to extraction and degradation of water-soluble components, pectin and ash. Thus the cellulose content increased by 6% w/w to 67-70% w/w. Steam explosion of the untreated hemp fibres increased the cellulose content to 74% w/w, whereas steam explosion of enzymatically treated hemp increased the cellulose content to 78% w/w. (author)

  2. A Longitudinal Investigation of the Affective Environment in Families with Young Children: From Infancy to Early School Age


    Barry, Robin A.; Kochanska, Grazyna


    We examined the affective environment in 102 families studied longitudinally when children were 7, 15, 25, 38, 52, and 67 months. At each assessment, every mother-child and father-child dyad was observed in typical daily contexts. Each person’s emotions of affection, joy, and anger were coded. Both parents rated marital quality when children were 15, 52, and 67 months. Growth curve analyses, using Actor-Partner Interdependence Modeling, examined (a) developmental changes in emotions, (b) with...

  3. In vitro zygotic embryo culture of Pinus peuce Gris.: Optimization of culture conditions affecting germination and early seedling growth

    Directory of Open Access Journals (Sweden)

    Stojičić Dragana


    Full Text Available This study reports a protocol for the germination and early seedling growth of Pinus peuce Gris. using zygotic embryo culture. In order to overcome seed dormancy and optimize organogenesis, the effect of nutritional, plant growth regulatory and physical factors on in vitro germination and growth of isolated mature zygotic embryos of P. peuce were investigated.

  4. Is the efficacy of hormonal therapy affected by lymph node status? data from the bicalutamide (Casodex) Early Prostate Cancer program

    DEFF Research Database (Denmark)

    Iversen, Peter; Wirth, Manfred P; See, William A;


    To report an exploratory subgroup analysis assessing the extent to which the overall benefit found in the Early Prostate Cancer program is dependent on lymph node status at randomization. The program is ongoing, and the overall survival data are immature. The first combined analysis of the bicalu...

  5. Does the Early Social Environment Affect Structure and Consistency of Personality in Wild-Type Male Rats?

    NARCIS (Netherlands)

    Gracceva, Giulia; Koolhaas, Jaap M.; Groothuis, Ton G. G.


    Animal personality has been extensively studied from a functional and evolutionary point of view. Less attention has been paid to the development of personality, its phenotypic plasticity, and the influence of manipulation of early environmental factors. Here we describe the effects of manipulating

  6. Recommendations for Implementing the New Illinois Early Learning and Development Standards to Affect Classroom Practices for Social and Emotional Learning (United States)

    Zinsser, Katherine M.; Dusenbury, Linda


    The state of Illinois in the central United States has long been a trendsetter both in the development of learning standards and in addressing social and emotional learning in education settings. With a recent revision to the state's early learning standards, published in 2013, the Illinois State Board of Education (ISBE) fully aligned its…

  7. How age and sex affect the erythrocyte sedimentation rate and C-reactive protein in early rheumatoid arthritis

    NARCIS (Netherlands)

    Siemons, L.; Klooster, P.M. ten; Vonkeman, H.E.; Riel, P.L.C.M. van; Glas, C.A.; Laar, M.A. van der


    BACKGROUND: The erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) are two commonly used measures of inflammation in rheumatoid arthritis (RA). As current RA treatment guidelines strongly emphasize early and aggressive treatment aiming at fast remission, optimal measurement of inflamm

  8. Difficulty in disengaging from threat and temperamental negative affectivity in early life: A longitudinal study of infants aged 12–36 months

    Directory of Open Access Journals (Sweden)

    Nakagawa Atsuko


    Full Text Available Abstract Background Attention disengagement is reportedly influenced by perceiving a fearful facial expression even in the first year of life. In the present study, we examined whether individual differences in disengaging from fearful expressions predict temperamental negative affectivity. Method Twenty-six infants were studied longitudinally at 12, 18, 24, and 36 months, using an overlap paradigm and two temperament questionnaires: the Japanese versions of the revised Infant Behavior Questionnaire and Early Childhood Behavior Questionnaire. Results The infants fixated significantly more frequently to fearful than to happy or neutral faces. The attentional bias to threat (i.e., the number of fixed responses on fearful faces divided by the total number of fixed responses on faces at 12 months was significantly positively correlated with negative affect at 12 months, and its relations with negative affect measured later in development was in the expected positive direction at each age. In addition, a moderation analysis indicates that the orienting network and not the executive network marginally moderated the relation between early attentional bias and later fear. Conclusions The results suggest that at 12 months, infants with more negative affectivity exhibit greater difficulty in disengaging their attention from fearful faces. We also found evidence that the association between parent-reported fear and disengagement might be modulated in the second year, perhaps because of the differences in temperamental control networks.

  9. [Enzymatic cholecystitis and cholecystopancreatitis]. (United States)

    Sobeshchanskaia, E A; Myrkin, S D; Babalich, A K


    Taking into consideration the role of pancreatic enzymes in the etiology of cholecystitis and cholecystopancreatits, the authors examined amylase in 92 patients, bile lipase--in 39 patients during cholecystectomy and also amylase in the choledochus bile during external drainage of the duct in 15 patients in the early postoperative period. The investigations inducated different levels of pancreatic enzymes in bile. Their level is found to depend on the occurrence of hypertension in the bile tract both pre- and postoperatively.

  10. Multi-instrumental Investigation of Affecting of Early Somatic Embryos of Spruce by Cadmium(II and Lead(II Ions

    Directory of Open Access Journals (Sweden)

    René Kizek


    Full Text Available The main aim of this work was to use multi-instrumental analytical apparatus toinvestigate the effects of treatment with cadmium(II and/or lead(II ions (50, 250 and 500μM for twelve days on early somatic spruce embryos (ESEs. Primarily we used imageanalysis for estimation of growth and a fluorimetric sensor for enzymatic detection ofviability of the treated ESEs. It follows from the obtained results that Cd caused highertoxicity to ESEs than Pb. Besides this fundamental finding, we observed that ESEs grewand developed better in the presence of 500 μM of the metal ions than in the presence of250 μM. Based on the results obtained using nuclear magnetic resonance this phenomenonwas related to an increase of the area of ESE clusters by intensive uptake of water from thecultivation medium, due to dilution of the heavy metal concentration inside the cluster. Inaddition we studied the glutathione content in treated ESEs by the adsorptive transferstripping technique coupled with the differential pulse voltammetry Brdicka reaction. GSHcontents increased up to 148 ng/mg (clone 2/32 and 158 ng/mg (clone PE 14 after twelve day long treatment with Cd-EDTA ions. The GSH content was about 150 and 160 % higher in comparison with the ESEs treated with Pb-EDTA ions, respectively. The difference between GSH contents determined in ESEs treated with Pb-EDTA and Cd-EDTA ions correlates with the higher toxicity of cadmium(II ions.

  11. Family Routines and Parental Monitoring as Protective Factors among Early and Middle Adolescents Affected by Maternal HIV/AIDS (United States)

    Murphy, Debra A.; Marelich, William D.; Herbeck, Diane M.; Payne, Diana L.


    The influence of parenting skills on adolescent outcomes among children affected by maternal HIV/AIDS (N = 118, M age = 13) was investigated. Among families with more frequent family routines, over time adolescents showed lower rates of aggression, anxiety, worry, depression, conduct disorder, binge drinking, and increased self-concept. Among…

  12. Enzymatically Controlled Vacancies in Nanoparticle Crystals

    Energy Technology Data Exchange (ETDEWEB)

    Barnaby, Stacey N.; Ross, Michael B.; Thaner, Ryan V.; Lee, Byeongdu; Schatz, George C.; Mirkin, Chad A.


    In atomic systems, the mixing of metals results in distinct phase behavior that depends on the identity and bonding characteristics of the atoms. In nanoscale systems, the use of oligonucleotides as programmable “bonds” that link nanoparticle “atoms” into superlattices allows for the decoupling of atom identity and bonding. While much research in atomic systems is dedicated to understanding different phase behavior of mixed metals, it is not well understood on the nanoscale how changes in the nanoscale “bond” affect the phase behavior of nanoparticle crystals. In this work, the identity of the atom is kept the same but the chemical nature of the bond is altered, which is not possible in atomic systems, through the use of DNA and RNA bonding elements. These building blocks assemble into single crystal nanoparticle superlattices with mixed DNA and RNA bonding elements throughout. The nanoparticle crystals can be dynamically changed through the selective and enzymatic hydrolysis of the RNA bonding elements, resulting in superlattices that retain their crystalline structure and habit, while incorporating up to 35% random vacancies generated from the nanoparticles removed. Therefore, the bonding elements of nanoparticle crystals can be enzymatically and selectively addressed without affecting the nature of the atom.

  13. Effects of organic carbon sequestration strategies on soil enzymatic activities (United States)

    Puglisi, E.; Suciu, N.; Botteri, L.; Ferrari, T.; Coppolecchia, D.; Trevisan, M.; Piccolo, A.


    Greenhouse gases emissions can be counterbalanced with proper agronomical strategies aimed at sequestering carbon in soils. These strategies must be tested not only for their ability in reducing carbon dioxide emissions, but also for their impact on soil quality: enzymatic activities are related to main soil ecological quality, and can be used as early and sensitive indicators of alteration events. Three different strategies for soil carbon sequestration were studied: minimum tillage, protection of biodegradable organic fraction by compost amendment and oxidative polimerization of soil organic matter catalyzed by biometic porfirins. All strategies were compared with a traditional agricultural management based on tillage and mineral fertilization. Experiments were carried out in three Italian soils from different pedo-climatic regions located respectively in Piacenza, Turin and Naples and cultivated with maize or wheat. Soil samples were taken for three consecutive years after harvest and analyzed for their content in phosphates, ß-glucosidase, urease and invertase. An alteration index based on these enzymatic activities levels was applied as well. The biomimetic porfirin application didn't cause changes in enzymatic activities compared to the control at any treatment or location. Enzymatic activities were generally higher in the minimum tillage and compost treatment, while differences between location and date of samplings were limited. Application of the soil alteration index based on enzymatic activities showed that soils treated with compost or subjected to minimum tillage generally have a higher biological quality. The work confirms the environmental sustainability of the carbon sequestering agronomical practices studied.

  14. Disturbance of the gut microbiota in early-life selectively affects visceral pain in adulthood without impacting cognitive or anxiety-related behaviors in male rats. (United States)

    O'Mahony, S M; Felice, V D; Nally, K; Savignac, H M; Claesson, M J; Scully, P; Woznicki, J; Hyland, N P; Shanahan, F; Quigley, E M; Marchesi, J R; O'Toole, P W; Dinan, T G; Cryan, J F


    Disruption of bacterial colonization during the early postnatal period is increasingly being linked to adverse health outcomes. Indeed, there is a growing appreciation that the gut microbiota plays a role in neurodevelopment. However, there is a paucity of information on the consequences of early-life manipulations of the gut microbiota on behavior. To this end we administered an antibiotic (vancomycin) from postnatal days 4-13 to male rat pups and assessed behavioral and physiological measures across all aspects of the brain-gut axis. In addition, we sought to confirm and expand the effects of early-life antibiotic treatment using a different antibiotic strategy (a cocktail of pimaricin, bacitracin, neomycin; orally) during the same time period in both female and male rat pups. Vancomycin significantly altered the microbiota, which was restored to control levels by 8 weeks of age. Notably, vancomycin-treated animals displayed visceral hypersensitivity in adulthood without any significant effect on anxiety responses as assessed in the elevated plus maze or open field tests. Moreover, cognitive performance in the Morris water maze was not affected by early-life dysbiosis. Immune and stress-related physiological responses were equally unaffected. The early-life antibiotic-induced visceral hypersensitivity was also observed in male rats given the antibiotic cocktail. Both treatments did not alter visceral pain perception in female rats. Changes in visceral pain perception in males were paralleled by distinct decreases in the transient receptor potential cation channel subfamily V member 1, the α-2A adrenergic receptor and cholecystokinin B receptor. In conclusion, a temporary disruption of the gut microbiota in early-life results in very specific and long-lasting changes in visceral sensitivity in male rats, a hallmark of stress-related functional disorders of the brain-gut axis such as irritable bowel disorder.

  15. Pollen production in sunflower (Helianthus annuus L.) is affected by air temperature and relative humidity during early reproductive growth


    Astiz, V.; Hernández, Luis Francisco


    During microsporogenesis, sunflower florets might be exposed to episodes of day- and night-time temperatures exceeding 30 and 20 °C, respectively, that can affect pollen availability and consequently plant’s yield. The relationship between air temperature and relative humidity during the meiotic phase of microsporogenesis, and the quantity and quality of pollen produced by the flowers, were studied in two sunflower hybrids during two years. The hybrids were grown on irrigated plots in Bahía B...

  16. TGF-β1 expression in wound healing is acutely affected by experimental malnutrition and early enteral feeding. (United States)

    Alves, Claudia Cristina; Torrinhas, Raquel Susana; Giorgi, Ricardo; Brentani, Maria Mitzi; Logullo, Angela Flavia; Waitzberg, Dan Linetzky


    Malnutrition is associated with the delay or failure of healing. We assessed the effect of experimental malnutrition and early enteral feeding with standard diet or diet supplemented with arginine and antioxidants on the levels of mRNA encoding growth factors in acute, open wound healing. Standardised cutaneous dorsal wounds and gastrostomies for enteral feeding were created in malnourished (M, n = 27) and eutrophic control (E, n = 30) Lewis male adult rats. Both M and E rats received isocaloric and isonitrogenous regimens with oral chow and saline (C), standard (S) or supplemented (A) enteral diets. On post-trauma day 7, mRNA levels of growth factor genes were analysed in wound granulation tissue by reverse transcription polymerase chain reaction (RT-PCR). M(C) rats had significantly lower transforming growth factor β(TGF-β1 ) mRNA levels than E(C) rats (2·58 ± 0·83 versus 3·53 ± 0·57, P malnutrition decreased local mRNA levels of TGF-β1 genes, which was minimised by early enteral feeding with standard or supplemented diets.

  17. Does the early social environment affect structure and consistency of personality in wild-type male's rat? (United States)

    Gracceva, Giulia; Koolhaas, Jaap M; Groothuis, Ton G G


    Animal personality has been extensively studied from a functional and evolutionary point of view. Less attention has been paid to the development of personality, its phenotypic plasticity, and the influence of manipulation of early environmental factors. Here we describe the effects of manipulating the sex ratio of the litter, at postnatal day (pnd) 3, in wild-type rats, on personality traits in adulthood. We measured the treatment effects on aggression, defensive burying, and open field behavior at pnd 90 and 120, as well as on their contextual generality, and stability over time (differential and structural consistency). Main effects of litter composition were found on open field behavior at pnd 120 but not on the other behaviors. Since correlations between behaviors changed over time irrespective of the specific treatment, whereas in previous studies on unmanipulated litters this was not the case we suggest that early handling may disrupt adult personality traits. Overall the data indicate that personality is less stable over time that often assumed, having both proximate and ultimate implications.

  18. Early environmental enrichment affects neurobehavioral development and prevents brain damage in rats submitted to neonatal hypoxia-ischemia. (United States)

    Schuch, Clarissa Pedrini; Diaz, Ramiro; Deckmann, Iohanna; Rojas, Joseane Jiménez; Deniz, Bruna Ferrary; Pereira, Lenir Orlandi


    Our previous results demonstrated improved cognition in adolescent rats housed in environmental enrichment (EE) that underwent neonatal hypoxia-ischemia (HI). The aim of this study was to investigate the effects of early EE on neurobehavioral development and brain damage in rats submitted to neonatal HI. Wistar rats were submitted to the HI procedure on the 7th postnatal day (PND) and housed in an enriched environment (8th-20th PND). The maturation of physical characteristics and the neurological reflexes were evaluated and the volume of striatum, corpus callosum and neocortex was measured. Data analysis demonstrated a clear effect of EE on neurobehavioral development; also, daily performance was improved in enriched rats on righting, negative geotaxis and cliff aversion reflex. HI caused a transient motor deficit on gait latency. Brain atrophy was found in HI animals and this damage was partially prevented by the EE. In conclusion, early EE stimulated neurobehavioral development in neonate rats and also protects the neocortex and the corpus callosum from atrophy following HI. These findings reinforce the potential of EE as a strategy for rehabilitation following neonatal HI and provide scientific support to the use of this therapeutic strategy in the treatment of neonatal brain injuries in humans.

  19. Enzymatic Browning: a practical class

    Directory of Open Access Journals (Sweden)

    Maria Teresa Pedrosa Silva Clerici


    Full Text Available This paper presents a practical class about the enzymes polyphenol oxidases, which have been shown to be responsible for the enzymatic browning of fruits and vegetables. Vegetables samples were submitted to enzymatic inactivation process with chemical reagents, as well as by bleaching methods of applying heat by conventional oven and microwave oven. Process efficiency was assessed qualitatively by both observing the guaiacol peroxidase activity and after the storage period under refrigeration or freezing. The practical results obtained in this class allow exploring multidisciplinary knowledge in food science, with practical applications in everyday life.

  20. A single early postnatal estradiol injection affects morphology and gene expression of the ovary and parametrial adipose tissue in adult female rats

    DEFF Research Database (Denmark)

    Alexanderson, Camilla; Stener-Victorin, Elisabet; Kullberg, Joel;


    expression related to follicular development and adipose tissue metabolism, and developed a non-invasive volumetric estimation of parametrial adipose tissue by magnetic resonance imaging. Estradiol reduced ovarian weight, increased antral follicle size and number of atretic antral follicles, and decreased......Events during early life can affect reproductive and metabolic functions in adulthood. We evaluated the programming effects of a single early postnatal estradiol injection (within 3h after birth) in female rats. We assessed ovarian and parametrial adipose tissue morphology, evaluated gene...... theca interna thickness in atretic antral follicles. Adult estradiol-injected rats also had malformed vaginal openings and lacked corpora lutea, confirming anovulation. Estradiol markedly reduced parametrial adipose tissue mass. Adipocyte size was unchanged, suggesting reduced adipocyte number...

  1. Ischemia in tumors induces early and sustained phosphorylation changes in stress kinase pathways but does not affect global protein levels

    Energy Technology Data Exchange (ETDEWEB)

    Mertins, Philipp; Yang, Feng; Liu, Tao; Mani, DR; Petyuk, Vladislav A.; Gillette, Michael; Clauser, Karl; Qiao, Jana; Gritsenko, Marina A.; Moore, Ronald J.; Levine, Douglas; Townsend, Reid; Erdmann-Gilmore, Petra; Snider, Jacqueline E.; Davies, Sherri; Ruggles, Kelly; Fenyo, David; Kitchens, R. T.; Li, Shunqiang; Olvera, Narcisco; Dao, Fanny; Rodriguez, Henry; Chan, Daniel W.; Liebler, Daniel; White, Forest; Rodland, Karin D.; Mills, Gordon; Smith, Richard D.; Paulovich, Amanda G.; Ellis, Matthew; Carr, Steven A.


    Advances in quantitative mass spectrometry (MS)-based proteomics have sparked efforts to characterize the proteomes of tumor samples to provide complementary and unique information inaccessible by genomics. Tumor samples are usually not accrued with proteomic characterization in mind, raising concerns regarding effects of undocumented sample ischemia on protein abundance and phosphosite stoichiometry. Here we report the effects of cold ischemia time on clinical ovarian cancer samples and patient-derived basal and luminal breast cancer xenografts. Tumor tissues were excised and collected prior to vascular ligation, subjected to accurately defined ischemia times up to 60 min, and analyzed by quantitative proteomics and phosphoproteomics using isobaric tags and high-performance, multidimensional LC-MS/MS. No significant changes were detected at the protein level in each tumor type after 60 minutes of ischemia, and the majority of the >25,000 phosphosites detected were also stable. However, large, reproducible increases and decreases in protein phosphorylation at specific sites were observed in up to 24% of the phosphoproteome starting as early as 5 minutes post-excision. Early and sustained activation of stress response, transcriptional regulation and cell death pathways were observed in common across tumor types. Tissue-specific changes in phosphosite stability were also observed suggesting idiosyncratic effects of ischemia in particular lineages. Our study provides insights into the information that may be obtained by proteomic characterization of tumor samples after undocumented periods of ischemia, and suggests caution especially in interpreting activation of stress pathways in such samples as they may reflect sample handling rather than tumor physiology.

  2. Strontium and neodymium isotopic variations in early Archean gneisses affected by middle to late Archean high-grade metamorphic processes: West Greenland and Labrador (United States)

    Collerson, K. D.; Mcculloch, M. T.; Bridgwater, D.; Mcgregor, V. R.; Nutman, A. P.


    Relicts of continental crust formed more than 3400 Ma ago are preserved fortuitously in most cratons. The cratons provide the most direct information about crust and mantle evolutionary processes during the first billion years of Earth history. In view of their polymetamorphic character, these terrains are commonly affected by subsequent tectonothermal events. Hence, their isotope systematics may be severely disturbed as a result of bulk chemical change or local isotopic homogenization. This leads to equivocal age and source information for different components within these terrains. The Sr and Nd isotopic data are presented for early Archean gneisses from the North Atlantic Craton in west Greenland and northern Labrador which were affected by younger metamorphic events.

  3. Bioluminescence methods for enzymatic determinations (United States)

    Bostick, William D.; Denton, Mark S.; Dinsmore, Stanley R.


    An enzymatic method for continuous, on-line and rapid detection of diagnostically useful biomarkers, which are symptomatic of disease or trauma-related tissue damage, is disclosed. The method is characterized by operability on authentic samples of complex biological fluids which contain the biomarkers.

  4. Rare Variants in PLD3 Do Not Affect Risk for Early-Onset Alzheimer Disease in a European Consortium Cohort. (United States)

    Cacace, Rita; Van den Bossche, Tobi; Engelborghs, Sebastiaan; Geerts, Nathalie; Laureys, Annelies; Dillen, Lubina; Graff, Caroline; Thonberg, Håkan; Chiang, Huei-Hsin; Pastor, Pau; Ortega-Cubero, Sara; Pastor, Maria A; Diehl-Schmid, Janine; Alexopoulos, Panagiotis; Benussi, Luisa; Ghidoni, Roberta; Binetti, Giuliano; Nacmias, Benedetta; Sorbi, Sandro; Sanchez-Valle, Raquel; Lladó, Albert; Gelpi, Ellen; Almeida, Maria Rosário; Santana, Isabel; Tsolaki, Magda; Koutroumani, Maria; Clarimon, Jordi; Lleó, Alberto; Fortea, Juan; de Mendonça, Alexandre; Martins, Madalena; Borroni, Barbara; Padovani, Alessandro; Matej, Radoslav; Rohan, Zdenek; Vandenbulcke, Mathieu; Vandenberghe, Rik; De Deyn, Peter P; Cras, Patrick; van der Zee, Julie; Sleegers, Kristel; Van Broeckhoven, Christine


    Rare variants in the phospholipase D3 gene (PLD3) were associated with increased risk for late-onset Alzheimer disease (LOAD). We identified a missense mutation in PLD3 in whole-genome sequence data of a patient with autopsy confirmed Alzheimer disease (AD) and onset age of 50 years. Subsequently, we sequenced PLD3 in a Belgian early-onset Alzheimer disease (EOAD) patient (N = 261) and control (N = 319) cohort, as well as in European EOAD patients (N = 946) and control individuals (N = 1,209) ascertained in different European countries. Overall, we identified 22 rare variants with a minor allele frequency <1%, 20 missense and two splicing mutations. Burden analysis did not provide significant evidence for an enrichment of rare PLD3 variants in EOAD patients in any of the patient/control cohorts. Also, meta-analysis of the PLD3 data, including a published dataset of a German EOAD cohort, was not significant (P = 0.43; OR = 1.53, 95% CI 0.60-3.31). Consequently, our data do not support a role for PLD3 rare variants in the genetic etiology of EOAD in European EOAD patients. Our data corroborate the negative replication data obtained in LOAD studies and therefore a genetic role of PLD3 in AD remains to be demonstrated.

  5. High intensity interval training favourably affects antioxidant and inflammation mRNA expression in early-stage chronic kidney disease. (United States)

    Tucker, Patrick S; Briskey, David R; Scanlan, Aaron T; Coombes, Jeff S; Dalbo, Vincent J


    Increased levels of oxidative stress and inflammation have been linked to the progression of chronic kidney disease. To reduce oxidative stress and inflammation related to chronic kidney disease, chronic aerobic exercise is often recommended. Data suggests high intensity interval training may be more beneficial than traditional aerobic exercise. However, appraisals of differing modes of exercise, along with explanations of mechanisms responsible for observed effects, are lacking. This study assessed effects of eight weeks of high intensity interval training (85% VO2max), versus low intensity exercise (45-50% VO2max) and sedentary behaviour, in an animal model of early-stage chronic kidney disease. We examined kidney-specific mRNA expression of genes related to endogenous antioxidant enzyme activity (glutathione peroxidase 1; Gpx1, superoxide dismutase 1; Sod1, and catalase; Cat) and inflammation (kidney injury molecule 1; Kim1 and tumour necrosis factor receptor super family 1b; Tnfrsf1b), as well as plasma F2-isoprostanes, a marker of lipid peroxidation. Compared to sedentary behaviour, high intensity interval training resulted in increased mRNA expression of Sod1 (p=0.01) and Cat (pinflammation.

  6. Analysis on enzymatic browning in pine needles

    Energy Technology Data Exchange (ETDEWEB)

    Kong, K.H.; Park, H.J.; Choi, S.S.; Cho, S.H. [Chung-Ang University, Seoul (Korea); Kim, Y.T. [Aoyama Gakuin University, Tokyo (Japan)


    Tyrosinases are related to the enzymatic browning of plants and attract the major scientific interest for the prevention of it. Three tyrosinase isozymes (P{sub 1}, P{sub 2} and P{sub 3}) from pine needles were purified to homogeneity and characterized the factors that affect their activities. The L-ascorbic acid and {beta}-mercaptoethanol notably inhibited the enzymatic activities of the three isozymes. The sodium diethyldithiocarbamate was a competitive inhibitor of isozymes with the K{sub i} values of P{sub 1}(0.30 mM), P{sub 2}(0.015 mM) and P{sub 3}(0.019 mM), respectively. Their enzyme activities were however, increased by the addition of most metal ions. The optimum pH for the three isozymes was 9.0{approx}9.5 and the optimum temperatures ranged from 55 to 60{sup o} C using L-DOPA as substrate. 15 refs., 3 figs., 2 tabs.

  7. Lactose hydrolysis in an enzymatic membrane reactor

    Energy Technology Data Exchange (ETDEWEB)

    Mertens, B.; Huyghebaert, A.


    The enzymatic hydrolysis of lactose in whey permeate with subsequent recuperation of Saccharomyces lactis lactase by means of ultrafiltration was investigated. In whey permeate, S. lactis lactase shows maximal activity at pH 6.5; the optimal temperature was found to be 45/sup 0/C and is limited by strong thermal inactivation beyond this temperature. High activity combined with acceptable thermal inactivation (< 10% after 5 h incubation) was established at 30/sup 0/C. S. lactis lactase also displays considerable activity at low temperature (5/sup 0/C). Enzyme stability is reduced drastically by demineralisation: addition of low concentrations of manganese ions (10/sup -3/ M) considerably enhances stability. Using a DDS Lab-Unit 35 fitted with GR61PP polysulphon membranes (cut-off: 20.000), pilot scale experiments were carried out (pH 6.5; 30/sup 0/C) in which whey permeate was hydrolyzed to a degree of hydrolysis of 82% minimum. Enzyme recuperation amounted to 96.5% per batch, all enzyme activity loss being due to thermal inactivation. Microbiological examination of the enzymatic membrane reactor showed that growth of mcicroorganisms can largely be suppressed by working at lower temperature (5/sup 0/C). Eventually, 50 ppm H/sub 2/O/sub 2/ or sterile filtration will adequately solve microbiological problems without affecting enzyme activity.

  8. Early visual processing is affected by clinical subtype in patients with unilateral spatial neglect: A magnetoencephalography study.

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    Katsuhiro eMizuno


    Full Text Available OBJECTIVE: To determine whether visual evoked fields (VEFs elicited by right and left hemifield stimulation differ in patients with unilateral spatial neglect that results from cerebrovascular accident.METHODS: Pattern-reversal stimulation of the right and left hemifield was performed in three patients with left unilateral spatial neglect. Magnetoencephalography was recorded using a 160-channel system, and VEFs were quantified in the 400 ms after each stimulus. The presence or absence of VEF components at around 100 ms (P100m component and 145 ms (N145m component after stimulus onset was determined. The source of the VEF was determined using a single equivalent current dipole model for spherical volume conduction. All patients were evaluated using the behavioral inattention test (BIT.RESULTS: In response to right hemifield stimulation, the P100m and N145m components of the VEF were evident in all three patients. In response to left hemifield stimulation, both components were evident in Patient 3, whereas only the P100m component was evident in Patient 1 and only the N145m component was evident in Patient 2. Patient 1 exhibited impairments on the line bisection and cancellation tasks of the BIT, Patient 2 exhibited impairments on the copying, drawing and cancellation tasks of the BIT, and Patient 3 exhibited impairments on the cancellation task of the BIT.CONCLUSIONS: These results demonstrate that early VEFs are disrupted in patients with unilateral spatial neglect and support the concept that deficits in visual processing differ according to the clinical subtype of unilateral spatial neglect and the lesion location. This study also demonstrates the feasibility of using magnetoencephalography to explore subtypes of neglect.

  9. Kinetic modelling of enzymatic starch hydrolysis

    NARCIS (Netherlands)

    Bednarska, K.A.


    Kinetic modelling of enzymatic starch hydrolysis – a summary K.A. Bednarska The dissertation entitled ‘Kinetic modelling of enzymatic starch hydrolysis’ describes the enzymatic hydrolysis and kinetic modelling of liquefaction and saccharification of wheat starch. A

  10. 2011 and 2012 Early Careers Achievement Awards: farm and pig factors affecting welfare during the marketing process. (United States)

    Johnson, A K; Gesing, L M; Ellis, M; McGlone, J J; Berg, E; Lonergan, S M; Fitzgerald, R; Karriker, L A; Ramirez, A; Stalder, K J; Sapkota, A; Kephart, R; Selsby, J T; Sadler, L J; Ritter, M J


    The objective of this paper is to review the scientific literature to identify on-farm factors that contribute to market weight pig transportation losses. Transportation of market weight pigs is an essential element to the multisite pork production model used in the United States. In 2011 alone, approximately 111 million market weight pigs were transported from the finishing site to the abattoir. For pigs, the marketing process can present a combination of potentially novel, physical, and/or unfamiliar experiences that can be stressful. If the pig cannot cope with these sequential and additive stressors, then an increased rate of transportation losses could occur with a detrimental effect on pork carcass value. Current yearly estimates for transport losses are 1 million pigs (1%). A variety of market weight pig and farm factors have been reported to detrimentally affect transportation losses. By understanding how pigs interact with their environment during marketing, researchers, producers, and personnel at the abattoir may begin to identify, prioritize, and attempt to minimize or eliminate these stressors. This process will ultimately decrease transportation losses, improve pork quality, and increase profitability.

  11. Mutations altering the gammaretrovirus endoproteolytic motif affect glycosylation of the envelope glycoprotein and early events of the virus life cycle

    Energy Technology Data Exchange (ETDEWEB)

    Argaw, Takele; Wilson, Carolyn A., E-mail:


    Previously, we found that mutation of glutamine to proline in the endoproteolytic cleavage signal of the PERV-C envelope (RQKK to RPKK) resulted in non-infectious vectors. Here, we show that RPKK results in a non-infectious vector when placed in not only a PERV envelope, but also the envelope of a related gammaretrovirus, FeLV-B. The amino acid substitutions do not prevent envelope precursor cleavage, viral core and genome assembly, or receptor binding. Rather, the mutations result in the formation of hyperglycosylated glycoprotein and a reduction in the reverse transcribed minus strand synthesis and undetectable 2-LTR circular DNA in cells exposed to vectors with these mutated envelopes. Our findings suggest novel functions associated with the cleavage signal sequence that may affect trafficking through the glycosylation machinery of the cell. Further, the glycosylation status of the envelope appears to impact post-binding events of the viral life cycle, either membrane fusion, internalization, or reverse transcription. - Highlights: • Env cleavage signal impacts infectivity of gammaretroviruses. • Non-infectious mutants have hyper-glycosylated envelope that bind target cells. • Non-infectious mutants have defects in the formation of the double-stranded DNA. • Env cleavage motif has functions beyond cleavage of the env precursor.

  12. Negative affectivity and EEG asymmetry interact to predict emotional interference on attention in early school-aged children. (United States)

    Solomon, Beylul; O'Toole, Laura; Hong, Melanie; Dennis, Tracy A


    Negative affectivity (NA) is a broad construct that has been associated with the development of psychopathologies, such as anxiety, and with exaggerated attention to threatening stimuli. EEG asymmetry reflects biological individual differences in emotional reactivity that may underlie the association between NA and attention to threat. The present study included a sample of 31 five-seven year olds (M age in months=74.39, SD=6.57) to test the hypothesis that greater NA, combined with greater right anterior and posterior asymmetries, predicts increased attention interference following threat stimuli. Children completed an executive attention task which presented task-irrelevant threat (angry) and non-threat (neutral) faces prior to each trial. EEG asymmetry was measured at baseline for anterior, anterior-temporal and posterior scalp regions and child NA was measured via maternal report. As predicted, children showing greater NA and greater right anterior-temporal asymmetry showed more attention interference following angry faces. Additionally, two trend-level effects emerged: children showing greater NA and greater left anterior-temporal asymmetry showed less attention interference following angry faces, and children showing greater NA and greater left posterior asymmetry showed less attention interference, but only following neutral faces. Discussion focuses on the utility of using EEG asymmetry in the study of temperament, attentional biases, and the biological processes by which temperament confers risk for psychopathology.

  13. Unstable Expression of Commonly Used Reference Genes in Rat Pancreatic Islets Early after Isolation Affects Results of Gene Expression Studies.

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    Lucie Kosinová

    Full Text Available The use of RT-qPCR provides a powerful tool for gene expression studies; however, the proper interpretation of the obtained data is crucially dependent on accurate normalization based on stable reference genes. Recently, strong evidence has been shown indicating that the expression of many commonly used reference genes may vary significantly due to diverse experimental conditions. The isolation of pancreatic islets is a complicated procedure which creates severe mechanical and metabolic stress leading possibly to cellular damage and alteration of gene expression. Despite of this, freshly isolated islets frequently serve as a control in various gene expression and intervention studies. The aim of our study was to determine expression of 16 candidate reference genes and one gene of interest (F3 in isolated rat pancreatic islets during short-term cultivation in order to find a suitable endogenous control for gene expression studies. We compared the expression stability of the most commonly used reference genes and evaluated the reliability of relative and absolute quantification using RT-qPCR during 0-120 hrs after isolation. In freshly isolated islets, the expression of all tested genes was markedly depressed and it increased several times throughout the first 48 hrs of cultivation. We observed significant variability among samples at 0 and 24 hrs but substantial stabilization from 48 hrs onwards. During the first 48 hrs, relative quantification failed to reflect the real changes in respective mRNA concentrations while in the interval 48-120 hrs, the relative expression generally paralleled the results determined by absolute quantification. Thus, our data call into question the suitability of relative quantification for gene expression analysis in pancreatic islets during the first 48 hrs of cultivation, as the results may be significantly affected by unstable expression of reference genes. However, this method could provide reliable information

  14. Enzymatic method (phospholipase B) for studies on cestode infection (Hymenolepis nana) and its suppression by chemotherapy

    Energy Technology Data Exchange (ETDEWEB)

    Ottolenghi, A.; Rowland, J.T.


    The phospholipase B activity of the small intestine of mice infected with Hymenolepis nana has been studied to determine its value as a laboratory test for the presence of parasites and the chemotherapeutic effects of antitapeworm drugs. Mice infected with 500 H. nana eggs were examined on the 21st day of infection and the phospholipase B content of homogenates of small intestine was determined using lysolecithin (6.6 x 10/sup -3/ M) as the substrate. Some of the infected animals were treated with niclosamide according to schedules and doses known to affect worm development. Presence or absence of parasites was verified by visual inspection of the intestinal content. The enzymatic and visual methods gave equivalent results in both infected-not treated and infected-treated groups. Special features of niclosamide action (relative refractoriness of the early parasitic forms, enhanced effect of multiple doses) have been confirmed. The enzymatic method is proposed as a procedure for laboratory testing in chemotherapeutic investigations.

  15. Insights into enzymatic thiamin catalysis


    Wikner, Christer


    Thiamin diphosphate, the biologically active form of vitamin B,, functions as a cofactor in various enzymes in the cell. The protein enhances the reactivity of the cofactor by binding it in a very specific manner. In this work, based upon information from the crystal structure, the mechanism of the thiamin dependent enzyme transketolase from yeast has been investigated by various methods. In enzymatic thiamin catalysis, the protein has three major tasks in the formation of a...

  16. Exposure time to caffeine affects heartbeat and cell damage-related gene expression of zebrafish Danio rerio embryos at early developmental stages. (United States)

    Abdelkader, Tamer Said; Chang, Seo-Na; Kim, Tae-Hyun; Song, Juha; Kim, Dong Su; Park, Jae-Hak


    Caffeine is white crystalline xanthine alkaloid that is naturally found in some plants and can be produced synthetically. It has various biological effects, especially during pregnancy and lactation. We studied the effect of caffeine on heartbeat, survival and the expression of cell damage related genes, including oxidative stress (HSP70), mitochondrial metabolism (Cyclin G1) and apoptosis (Bax and Bcl2), at early developmental stages of zebrafish embryos. We used 100 µm concentration based on the absence of locomotor effects. Neither significant mortality nor morphological changes were detected. We monitored hatching at 48 h post-fertilization (hpf) to 96 hpf. At 60 and 72 hpf, hatching decreased significantly (P caffeine treatment with no significant difference (P > 0.05). Heartbeats per minute were 110, 110 and 112 in control at 48, 72 and 96 hpf, respectively. Caffeine significantly increased heartbeat - 122 and 136 at 72 and 96 hpf, respectively. Quantitative RT-PCR showed significant up-regulation after caffeine exposure in HSP70 at 72 hpf; in Cyclin G1 at 24, 48 and 72 hpf; and in Bax at 48 and 72 hpf. Significant down-regulation was found in Bcl2 at 48 and 72 hpf. The Bax/Bcl2 ratio increased significantly at 48 and 72 hpf. We conclude that increasing exposure time to caffeine stimulates oxidative stress and may trigger apoptosis via a mitochondrial-dependent pathway. Also caffeine increases heartbeat from early phases of development without affecting the morphology and survival but delays hatching. Use of caffeine during pregnancy and lactation may harm the fetus by affecting the expression of cell-damage related genes.

  17. Isobaric Tagging-Based Quantification for Proteomic Analysis: A Comparative Study of Spared and Affected Muscles from mdx Mice at the Early Phase of Dystrophy.

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    Cintia Yuri Matsumura

    Full Text Available Duchenne muscular dystrophy (DMD is the most common childhood myopathy, characterized by muscle loss and cardiorespiratory failure. While the genetic basis of DMD is well established, secondary mechanisms associated with dystrophic pathophysiology are not fully clarified yet. In order to obtain new insights into the molecular mechanisms of muscle dystrophy during earlier stages of the disease, we performed a comparative proteomic profile of the spared extraocular muscles (EOM vs. affected diaphragm from the mdx mice, using a label based shotgun proteomic approach. Out of the 857 identified proteins, 42 to 62 proteins had differential abundance of peptide ions. The calcium-handling proteins sarcalumenin and calsequestrin-1 were increased in control EOM compared with control DIA, reinforcing the view that constitutional properties of EOM are important for their protection against myonecrosis. The finding that galectin-1 (muscle regeneration, annexin A1 (anti-inflammatory and HSP 47 (fibrosis were increased in dystrophic diaphragm provides novel insights into the mechanisms through which mdx affected muscles are able to counteract dystrophy, during the early stage of the disease. Overall, the shotgun technique proved to be suitable to perform quantitative comparisons between distinct dystrophic muscles and allowed the suggestion of new potential biomarkers and drug targets for dystrophinopaties.

  18. Dietary patterns in pregnancy and effects on nutrient intake in the Mid-South: the Conditions Affecting Neurocognitive Development and Learning in Early Childhood (CANDLE) study. (United States)

    Völgyi, Eszter; Carroll, Kecia N; Hare, Marion E; Ringwald-Smith, Karen; Piyathilake, Chandrika; Yoo, Wonsuk; Tylavsky, Frances A


    Dietary patterns are sensitive to differences across socio-economic strata or cultural habits and may impact programing of diseases in later life. The purpose of this study was to identify distinct dietary patterns during pregnancy in the Mid-South using factor analysis. Furthermore, we aimed to analyze the differences in the food groups and in macro- and micronutrients among the different food patterns. The study was a cross-sectional analysis of 1155 pregnant women (mean age 26.5 ± 5.4 years; 62% African American, 35% Caucasian, 3% Other; and pre-pregnancy BMI 27.6 ± 7.5 kg/m(2)). Using food frequency questionnaire data collected from participants in the Conditions Affecting Neurocognitive Development and Learning in Early Childhood (CANDLE) study between 16 and 28 weeks of gestation, dietary patterns were identified using factor analysis. Three major dietary patterns, namely, Healthy, Processed, and US Southern were identified among pregnant women from the Mid-South. Further analysis of the three main patterns revealed four mixed dietary patterns, i.e., Healthy-Processed, Healthy-US Southern, Processed-US Southern, and overall Mixed. These dietary patterns were different (p food items, macro- and micro nutrients and aligned across socioeconomic and racial groups. Our study describes unique dietary patterns in the Mid-South, consumed by a cohort of women enrolled in a prospective study examining the association of maternal nutritional factors during pregnancy that are known to affect brain and cognitive development by age 3.

  19. The street children of Manila are affected by early-in-life periodontal infection: description of a treatment modality: sea salt. (United States)

    Michel, J F; Michel, M G; Nadan, J; Nowzari, H


    Thousands of street children of Manila are affected by early-in-life oral infection. The aim of the present investigation was to evaluate the effectiveness of a sea-salt mouthrinse solution in street children of Manila affected by mild to severe forms of periodontal disease. These children were all in need of special protection: abandoned, abused, exploited, neglected, orphaned, poor. During 3 oral-health missions in 2003, 2004 and 2005, 617 abandoned children (5 to 13 year-old), received oral examination at a non-sectarian child-caring institution in Metro Manila (Virlanie Foundation) by calibrated examiners. A treatment based on what could be done was proposed: 1. Teaching of a precise tooth brushing technique with sea-salt, controlled and reinforced every two days for one week by calibrated health educators, 2. The application of sea-salt water mouthrinse (2.5 gram in 20 ml). Periodontal measurements were repeated at the end of each mission. All children returned to child-caring institution for the followup examinations. In 2003, 10 male and 11 female (n=21) were diagnosed with aggressive periodontitis. In 2009 and 2010, none was affected by aggressive periodontitis. For all patients, the gingival index decreased from 1.08 at the first mission to 1.04 at the end of the second mission and 0.98 at the end of the third mission. The periodontal index decreased from 1.33 at the first mission to 0.98 at the second mission and 0.92 at the last mission. The present investigation confirms that prevention and early diagnosis can result in success with minimum cost. The provided oral health program empowered street children in the most desperate circumstances to be educated and become self-reliant, independent, and responsible. We propose here an antimicrobial approach which has a high degree of efficacy and tolerability, and can be implemented in virtually all parts of the world using low-cost resources.

  20. Effect of inhibitors released during steam-explosion pretreatment of barley straw on enzymatic hydrolysis. (United States)

    García-Aparicio, Ma Prado; Ballesteros, Ignacio; González, Alberto; Oliva, José Miguel; Ballesteros, Mercedes; Negro, Ma José


    The influence of the liquid fraction (prehydrolysate) generated during steam-explosion pretreatment (210 degrees C, 15 min) of barley straw on the enzymatic hydrolysis was determined. Prehydrolysate was analyzed for degradation compounds and sugars' content and used as a medium for enzymatic hydrolysis tests after pH adjusting to 4.8. Our results show that the presence of the compounds contained in the prehydrolysate strongly affects the hydrolysis step (a 25% decrease in cellulose conversion compared with control). Sugars are shown to be more potent inhibitors of enzymatic hydrolysis than degradation products.

  1. Enzymatic hydrolysis of potato pulp

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    Mariusz Lesiecki


    Full Text Available Background. Potato pulp constitutes a complicated system of four types of polysaccharides: cellulose, hemicellulose, pectin and starch. Its composition makes it a potential and attractive raw material for the production of the second generation bioethanol. The aim of this research project was to assess the usefulness of commercial enzymatic preparations for the hydrolysis of potato pulp and to evaluate the effectiveness of hydrolysates obtained in this way as raw materials for ethanol fermentation. Material  and methods. Sterilised potato pulp was subjected to hydrolysis with commercial enzymatic preparations. The effectiveness of the preparations declared as active towards only one fraction of potato pulp (separate amylase, pectinase and cellulase activity and mixtures of these preparations was analysed. The monomers content in hydrolysates was determined using HPLC method. Results.  The application of amylolytic enzymes for potato pulp hydrolysis resulted in the release of only 18% of raw material with glucose as the dominant (77% constituent of the formed product. In addition, 16% galactose was also determined in it. The hydrolysis of the cellulose fraction yielded up to 35% raw material and the main constituents of the obtained hydrolysate were glucose (46% and arabinose (40%. Simultaneous application of amylolytic, cellulolytic and pectinolytic enzymes turned out to be the most effective way of carrying out the process as its efficiency in this case reached 90%. The obtained hydrolysate contained 63% glucose, 25% arabinose and 12% other simple substances. Conclusion. The application of commercial enzymatic preparations made it possible to perform potato pulp hydrolysis with 90% effectiveness. This was achieved by the application of a complex of amylolytic, cellulolytic and pectinolytic enzymes and the hydrolysate obtained in this way contained, primarily, glucose making it a viable substrate for ethanol fermentation.

  2. Enzymatic activity of myccorrhizal fungi

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    Roman Pachlewski


    Full Text Available The investigations included assays of enzymatic activity of ectomycorrhizal fungi from the genera: Amanita, Cenococcum, Coltricia, Hebeloma, Lactarius, Rhizopogon, Russula, Suillus, Tricholoma and the pine ectendomycorrhizal strain MrgX. Among the 22 investigated strains of fungi 18 could decompose starch, 14 urea, 11 asparagine, 7 protein, 6 pectin and 3 ce1lulose. The most varied enzyme activities were found in Amanita muscaria, A. verna, Hebeloma, mesophaeum, ectendomycorrhizal isolate MrgX, Rhizopogon luteolus and Suillus bovinus, the highest cellolotytic activity was shown by the ectendomycorrhizal strain.


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    Wilberg K. Q.


    Full Text Available This paper presents an enzymatic process for sorbitol and gluconic acid production using cells of Zymomonas mobilis permeabilized with CTAB. Equimolar solutions of glucose and fructose (from 96.0 to 422.2 g/L were used. In a batch reactor, conversions of 97% were attained after 15 to 20 hours of reaction. The effect of the initial concentration of the substrates was evaluated in experiments using 20% more and 20% less glucose than fructose. It was observed that the reaction performed with more fructose reached completion faster and with a higher value of conversion

  4. Enzymatic profile of Haemophilus ducreyi

    Energy Technology Data Exchange (ETDEWEB)

    Casin, I.M.; Sanson-Le Pors, M.J.; Gorce, M.F.; Ortenberg, M.; Perol, Y. (Universite Paris - 7, Hopital Saint-Louis, 75 - Paris (France))

    The enzymatic activities of two reference strains of Haemophilus ducreyi and thirty clinical isolates were investigated by conventional biochemical tests and the API-ZYM test kit system which included 97 synthetic substrates. No strains converted acid to porphyrins, but they all reduced nitrates to nitrites. All strains possessed aminopeptidase activity against ..beta..-naphthylamide derivatives of L-alanine, L-arginine, L-glutamine, glycine, L-leucine, L-lysine and L-serine. No trypsin or chymotrypsin-like activities were detected. All strains had phosphatase activity with broad pH range, and phosphoamidase activity. No glycosidase was detected by the substrates tested.

  5. Effect of Steam Explosion Pretreatment on Bamboo for Enzymatic Hydrolysis and Ethanol Fermentation


    Zhiqiang Li; Benhua Fei; Zehui Jiang


    Based on the steam explosion pretreatment that has been applied to other types of lignocellulosic biomass, the steam explosion pretreatment of bamboo, along with a study of the chemical compositions and enzymatic hydrolyzability of substrates, was conducted. The results show that steam explosion pretreatment can greatly enhance the cellulose-to-glucose conversion yield after enzymatic hydrolysis, which is sometimes affected by bamboo age and steam explosion conditions. When the steam explosio...

  6. Early social instability affects plasma testosterone during adolescence but does not alter reproductive capacity or measures of stress later in life. (United States)

    Siegeler, Katja; Wistuba, Joachim; Damm, Oliver S; von Engelhardt, Nikolaus; Sachser, Norbert; Kaiser, Sylvia


    The social environment plays an important role in modulating processes of the hormonal and behavioural profile of an animal in a variety of group-living species. In wild cavies for instance, unstable social environmental conditions during pregnancy and lactation lead to an infantilised biobehavioural profile of the male offspring. In the present study, the influence of the social environment during pregnancy and lactation on the male wild cavy offsprings' plasma testosterone development, reproductive capacity and stress system activity was investigated. To this purpose, 12 sons whose mothers had lived in an unstable social environment during pregnancy and lactation were compared with 12 sons whose mothers had lived in a stable social environment during the same time. Plasma testosterone (T) and plasma cortisol (C) concentrations were determined from days 20 to 107 of age. Adrenal tyrosine hydroxylase (TH) activity and different parameters of reproductive capacity (weights of testes, epididymides and accessory sex glands, cellular composition of the testes, DNA fragmentation indices and sperm motility parameters) were analysed at day 107 of age. TH activity and plasma C were unaffected by different social environmental conditions early in life. The developmental time course of T concentrations, however, was significantly different: Sons whose mothers had lived in an unstable social environment during pregnancy and lactation showed a delayed increase in T concentrations around adolescence compared to controls. In contrast, no reproduction-related parameters measured within this study differed significantly between the two groups. Thus, early social instability affects plasma testosterone development during adolescence in a significant way but does not alter reproductive capacity or measures of stress later in life.

  7. Factors affecting spontaneous reduction of corpora lutea and twin embryos during the late embryonic/early fetal period in multiple-ovulating dairy cows. (United States)

    López-Gatius, F; García-Ispierto, I; Hunter, R H F


    Spontaneous reduction of advanced twin embryos has been described in high-producing, Holstein-Fresian (Bos taurus) dairy herds. The first objective of the current study was to determine whether management and cow factors could have an effect on such a reduction in twin pregnancies during the early fetal period. Because loss of a corpus luteum was noted in cows suffering twin reduction, we expanded our study to include multiple-ovulating cows carrying singletons. Pregnancy was diagnosed and confirmed from Days 28 to 34 and 56 to 62 postinsemination. Sixty-nine (23.5%) of 293 pregnant cows with two corpora lutea carrying singletons and 132 (28.4%) of 464 twin pregnancies recorded on first pregnancy diagnosis subsequently lost one of the corpora lutea or one of the embryos, respectively. Thirty-four (25.8%) of the 132 twin pregnancies suffering embryo reduction lost one corpus luteum along with the embryo. Corpus luteum reduction always occurred in the ovary ipsilateral to the gravid horn suffering embryo reduction. Binary logistic regressions were performed considering corpus luteum and embryo reduction as dependent variables in single and twin pregnancies, respectively, and several management- and cow-related factors as independent variables. In cows carrying singletons, the risk of corpus luteum reduction was 14.3 (1/0.07) times lower for a given herd, whereas the interaction season by laterality significantly affected corpus luteum reduction such that in cows with two corpora lutea ipsilateral to the horn of pregnancy, the risk of reduction decreased during the winter period. In cows carrying twins, ipsilateral twin pregnancies were 3.45 (1/0.29) times more likely to undergo the loss of one embryo than bilateral twin pregnancies. As an overall conclusion, both corpora lutea and embryos were vulnerable to the effects of stress factors during the early fetal period in cows maintaining their pregnancies. A strong unilateral relationship between the corpus luteum and

  8. The Autophagic Process Occurs in Human Bone Metastasis and Implicates Molecular Mechanisms Differently Affected by Rab5a in the Early and Late Stages

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    Paola Maroni


    Full Text Available Autophagy favours metastatic growth through fuelling energy and nutrients and resistance to anoikis, typical of disseminated-tumour cells. The autophagic process, mediated by a unique organelle, the autophagosome, which fuses with lysosomes, is divided into three steps. Several stages, especially early omegasome formation and isolation-membrane initiation, remain controversial; molecular mechanisms involve the small-GTPase Rab5a, which regulates vesicle traffic for autophagosome formation. We examined Rab5a involvement in the function of key members of ubiquitin-conjugation systems, Atg7 and LC3-lipidated, interacting with the scaffold-protein p62. Immunohistochemistry of Rab5a was performed in human specimens of bone metastasis and pair-matched breast carcinoma; the autophagic-molecular mechanisms affected by Rab5a were evaluated in human 1833 bone metastatic cells, derived from breast-carcinoma MDA-MB231 cells. To clarify the role of Rab5a, 1833 cells were transfected transiently with Rab5a-dominant negative, and/or stably with the short-hairpin RNA Atg7, were exposed to two inhibitors of autolysosome function, and LC3II and p62 expression was measured. We showed basal autophagy in bone-metastatic cells and the pivotal role of Rab5a together with Beclin 1 between the early stages, elongation of isolation membrane/closed autophagosome mediated by Atg7, and the late-degradative stages. This regulatory network might occur in bone-metastasis and in high-grade dysplastic lesions, preceding invasive-breast carcinoma and conferring phenotypic characteristics for dissemination.

  9. The Autophagic Process Occurs in Human Bone Metastasis and Implicates Molecular Mechanisms Differently Affected by Rab5a in the Early and Late Stages (United States)

    Maroni, Paola; Bendinelli, Paola; Resnati, Massimo; Matteucci, Emanuela; Milan, Enrico; Desiderio, Maria Alfonsina


    Autophagy favours metastatic growth through fuelling energy and nutrients and resistance to anoikis, typical of disseminated-tumour cells. The autophagic process, mediated by a unique organelle, the autophagosome, which fuses with lysosomes, is divided into three steps. Several stages, especially early omegasome formation and isolation-membrane initiation, remain controversial; molecular mechanisms involve the small-GTPase Rab5a, which regulates vesicle traffic for autophagosome formation. We examined Rab5a involvement in the function of key members of ubiquitin-conjugation systems, Atg7 and LC3-lipidated, interacting with the scaffold-protein p62. Immunohistochemistry of Rab5a was performed in human specimens of bone metastasis and pair-matched breast carcinoma; the autophagic-molecular mechanisms affected by Rab5a were evaluated in human 1833 bone metastatic cells, derived from breast-carcinoma MDA-MB231 cells. To clarify the role of Rab5a, 1833 cells were transfected transiently with Rab5a-dominant negative, and/or stably with the short-hairpin RNA Atg7, were exposed to two inhibitors of autolysosome function, and LC3II and p62 expression was measured. We showed basal autophagy in bone-metastatic cells and the pivotal role of Rab5a together with Beclin 1 between the early stages, elongation of isolation membrane/closed autophagosome mediated by Atg7, and the late-degradative stages. This regulatory network might occur in bone-metastasis and in high-grade dysplastic lesions, preceding invasive-breast carcinoma and conferring phenotypic characteristics for dissemination. PMID:27023526

  10. Low pH affects survival, growth, size distribution, and carapace quality of the postlarvae and early juveniles of the freshwater prawn Macrobrachium rosenbergii de Man (United States)

    Kawamura, Gunzo; Bagarinao, Teodora; Yong, Annita Seok Kian; Chen, Chiau Yu; Noor, Siti Norasidah Mat; Lim, Leong Seng


    Acidification of rain water caused by air pollutants is now recognized as a serious threat to aquatic ecosystems. We examined the effects of low pH (control pH 7.5, pH 6, pH 5, pH 4) on the survival, growth, and shell quality of Macrobrachium rosenbergii postlarvae and early juveniles in the laboratory. Hatcheryproduced postlarvae (PL 5) were stocked at 250 PL per aquarium, acclimated over 7 d to experimental pH adjusted with hydrochloric acid, and reared for 30 d. Dead specimens were removed and counted twice a day. After 27 d rearing, all specimens were measured for total length and body weight. Carapace quality was assessed by spectrophotometry. Survival of juveniles was highest at pH 6 (binomial 95% confidence interval 79 - 89%) followed by control pH 7.5 (56 - 68%) and pH 5 (50 - 60%) and was lowest for unmetamorphosed postlarvae and juveniles at pH 4 (43 - 49%). The final median total length and body weight of juveniles were similar at control pH 7.5 (18.2 TL, 50.2 mg BW) and pH 6 (17.7 mm TL, 45.0 mg BW) but significantly less at pH 5 (16.7 mm TL, 38.2 mg BW); at pH 4, the postlarvae did not metamorphose and measured only 9.8 mm TL, 29.3 mg BW. Length frequency distribution showed homogeneous growth at pH 6, positive skew at control pH 7.5 and pH 5, and extreme heterogeneity at pH 4. The carapace showed different transmittance spectra and lower total transmittance (i.e. thicker carapace) in juveniles at pH 7.5, pH 6, and pH 5 than in unmetamorphosed postlarvae and juveniles with thinner carapace at pH 4. Thus, survival, growth, size distribution, and carapace quality of M. rosenbergii postlarvae and early juveniles were negatively affected by pH 5 and especially pH 4. The thinner carapace of the survivors at pH 4 was mostly due to their small size and failure to metamorphose. Natural waters affected by acid rain could decimate M. rosenbergii populations in the wild.

  11. Dietary Patterns in Pregnancy and Effects on Nutrient Intake in the Mid-South: The Conditions Affecting Neurocognitive Development and Learning in Early Childhood (CANDLE Study

    Directory of Open Access Journals (Sweden)

    Frances A. Tylavsky


    Full Text Available Dietary patterns are sensitive to differences across socio-economic strata or cultural habits and may impact programing of diseases in later life. The purpose of this study was to identify distinct dietary patterns during pregnancy in the Mid-South using factor analysis. Furthermore, we aimed to analyze the differences in the food groups and in macro- and micronutrients among the different food patterns. The study was a cross-sectional analysis of 1155 pregnant women (mean age 26.5 ± 5.4 years; 62% African American, 35% Caucasian, 3% Other; and pre-pregnancy BMI 27.6 ± 7.5 kg/m2. Using food frequency questionnaire data collected from participants in the Conditions Affecting Neurocognitive Development and Learning in Early Childhood (CANDLE study between 16 and 28 weeks of gestation, dietary patterns were identified using factor analysis. Three major dietary patterns, namely, Healthy, Processed, and US Southern were identified among pregnant women from the Mid-South. Further analysis of the three main patterns revealed four mixed dietary patterns, i.e., Healthy-Processed, Healthy-US Southern, Processed-US Southern, and overall Mixed. These dietary patterns were different (p < 0.001 from each other in almost all the food items, macro- and micro nutrients and aligned across socioeconomic and racial groups. Our study describes unique dietary patterns in the Mid-South, consumed by a cohort of women enrolled in a prospective study examining the association of maternal nutritional factors during pregnancy that are known to affect brain and cognitive development by age 3.

  12. Real-time ESI-MS of enzymatic conversion: impact of organic solvents and multiplexing. (United States)

    Scheerle, Romy K; Grassmann, Johanna; Letzel, Thomas


    Different enzymatic assays were characterized systematically by real-time electrospray ionization mass spectrometry (ESI-MS) in the presence of organic solvents as well as in multiplex approaches and in a combination of both. Typically, biological enzymatic reactions are studied in aqueous solutions, since most enzymes show their full activity solely in aqueous solutions. However, in recent years, the use of organic solvents in combination with enzymatic reactions has gained increasing interest due to biotechnological advantages in chemical synthesis, development of online coupled setups screening for enzyme regulatory compounds, advantages regarding mass spectrometric detection and others. In the current study, the influence of several common organic solvents (methanol, ethanol, isopropanol, acetone, acetonitrile) on enzymatic activity (hen egg white lysozyme, chitinase, α-chymotrypsin, elastase from human neutrophils and porcine pancreas, acetylcholinesterase) was tested. Moreover, multiplexing is a promising approach enabling fast and cost-efficient screening methods, e.g. for determination of inhibitors in complex mixtures or in the field of biomedical research. Although in multiplexed setups the enzymatic activity may be affected by the presence of other substrates and/or enzymes, the expected advantages possibly will predominate. To investigate those effects, we measured multiple enzymatic assays simultaneously. For all conducted measurements, the conversion rate of the substrate(s) was calculated, which reflects the enzymatic activity. The results provide an overview about the susceptibility of the selected enzymes towards diverse factors and a reference point for many applications in analytical chemistry and biotechnology.

  13. Monitoring enzymatic ATP hydrolysis by EPR spectroscopy. (United States)

    Hacker, Stephan M; Hintze, Christian; Marx, Andreas; Drescher, Malte


    An adenosine triphosphate (ATP) analogue modified with two nitroxide radicals is developed and employed to study its enzymatic hydrolysis by electron paramagnetic resonance spectroscopy. For this application, we demonstrate that EPR holds the potential to complement fluorogenic substrate analogues in monitoring enzymatic activity.

  14. Suppression of microRNA-31 increases sensitivity to 5-FU at an early stage, and affects cell migration and invasion in HCT-116 colon cancer cells

    Directory of Open Access Journals (Sweden)

    Sun Xiao-Feng


    Full Text Available Abstract Background MicroRNAs (miRNAs are endogenously expressed noncoding RNAs with important biological and pathological functions. Although several studies have shown that microRNA-31 (miR-31 is obviously up-regulated in colorectal cancer (CRC, there is no study on the functional roles of miR-31 in CRC. Methods Anti-miR™ miRNA 31 inhibitor (anti-miR-31 is a sequence-specific and chemically modified oligonucleotide to specifically target and knockdown miR-31 molecule. The effect of anti-miR-31 transfection was investigated by real-time PCR. HCT-116p53+/+ and HCT-116p53-/-colon cancer cells were treated by anti-miR-31 with or without 5-fluorouracil (5-FU, cell proliferation was determined by MTT assay; apoptosis was detected by DAPI staining; cell cycle was evaluated by flow cytometry; colony formation, migration and invasion assays were performed to investigate the effect of suppression of miR-31 on the cell lines. Results Real-time PCR results showed that anti-miR-31 was efficiently introduced into the cells and reduced miR-31 levels to 44.1% in HCT-116p53+/+ and 67.8% in HCT-116p53-/-cell line (p = 0.042 and 0.046. MTT results showed that anti-miR-31 alone had no effect on the proliferation of HCT-116p53+/+ or HCT-116p53-/-. However, when combined with 5-FU, anti-miR-31 inhibited the proliferation of the two cell lines as early as 24 h after exposure to 5-FU (p = 0.038 and 0.044. Suppression of miR-31 caused a reduction of the migratory cells by nearly 50% compared with the negative control in both HCT-116p53+/+ and HCT-116p53-/-(p = 0.040 and 0.001. The invasive ability of the cells were increased by 8-fold in HCT-116p53+/+ and 2-fold in HCT-116p53-/- (p = 0.045 and 0.009. Suppression of miR-31 had no effect on cell cycle and colony formation (p > 0.05. Conclusions Suppression of miR-31 increases sensitivity to 5-FU at an early stage, and affects cell migration and invasion in HCT-116 colon cancer cells.

  15. Rapid near infrared spectroscopy for prediction of enzymatic hydrolysis of corn bran after various pretreatments

    DEFF Research Database (Denmark)

    Baum, Andreas; Agger, Jane; Meyer, Anne S.


    step is to increase the responsivity of the substrate to enzymatic attack and the type of pretreatment affects the enzymatic conversion efficiency. Destarched corn bran is a fibrous, heteroxylan-rich side-stream from the starch industry which may be used as a feedstock for bioethanol production...... or as a source of xylose for other purposes. In the present study we demonstrate the use of diffuse reflectance near infrared spectroscopy (NIR) as a rapid and non-destructive analytical tool for evaluation of pretreatment effects on destarched corn bran. NIR was used to achieve classification between 43...... differently pretreated corn bran samples using principal component analysis (PCA) and hierarchal clustering algorithms. Quantification of the enzymatically released monosaccharides by HPLC was used to design multivariate calibration models (biPLS) on the NIR spectra. The models could predict the enzymatic...

  16. Exploring crystalline-structural variations of cellulose during alkaline pretreatment for enhanced enzymatic hydrolysis. (United States)

    Ling, Zhe; Chen, Sheng; Zhang, Xun; Xu, Feng


    The study aimed to explore the crystallinity and crystalline structure of alkaline pretreated cellulose. The enzymatic hydrolysis followed by pretreatment was conducted for measuring the efficiency of sugar conversion. For cellulose Iβ dominated samples, alkaline pretreatment (cellulose crystallinity and depolymerized hemicelluloses, that were superimposed to affect the enzymatic conversion to glucose. Varying crystallite sizes and lattice spacings indicated the separation of cellulose crystals during mercerization (8-12wt% NaOH). Completion of mercerization was proved under higher alkaline concentration (14-18wt% NaOH), leading to distortion of crystalline cellulose to some extent. Cellulose II crystallinity showed a stimulative impact on enzymatic hydrolysis due to the weakened hydrophobic interactions within cellulose chains. The current study may provide innovative explanations for enhanced enzymatic digestibility of alkaline pretreated lignocellulosic materials.

  17. The Varicella-Zoster Virus Immediate-Early 63 protein affects chromatin controlled gene transcription in a cell-type dependent manner

    Directory of Open Access Journals (Sweden)

    Bontems Sébastien


    Full Text Available Abstract Background Varicella Zoster Virus Immediate Early 63 protein (IE63 has been shown to be essential for VZV replication, and critical for latency establishment. The activity of the protein as a transcriptional regulator is not fully clear yet. Using transient transfection assays, IE63 has been shown to repress viral and cellular promoters containing typical TATA boxes by interacting with general transcription factors. Results In this paper, IE63 regulation properties on endogenous gene expression were evaluated using an oligonucleotide-based micro-array approach. We found that IE63 modulates the transcription of only a few genes in HeLa cells including genes implicated in transcription or immunity. Furthermore, we showed that this effect is mediated by a modification of RNA POL II binding on the promoters tested and that IE63 phosphorylation was essential for these effects. In MeWo cells, the number of genes whose transcription was modified by IE63 was somewhat higher, including genes implicated in signal transduction, transcription, immunity, and heat-shock signalling. While IE63 did not modify the basal expression of several NF-κB dependent genes such as IL-8, ICAM-1, and IκBα, it modulates transcription of these genes upon TNFα induction. This effect was obviously correlated with the amount of p65 binding to the promoter of these genes and with histone H3 acetylation and HDAC-3 removal. Conclusion While IE63 only affected transcription of a small number of cellular genes, it interfered with the TNF-inducibility of several NF-κB dependent genes by the accelerated resynthesis of the inhibitor IκBα.

  18. The tuberculin skin test (TST is affected by recent BCG vaccination but not by exposure to non-tuberculosis mycobacteria (NTM during early life.

    Directory of Open Access Journals (Sweden)

    Sarah Burl

    Full Text Available The tuberculin skin test (TST is widely used in TB clinics to aid Mycobacterium tuberculosis (M.tb diagnosis, but the definition and the significance of a positive test in very young children is still unclear. This study compared the TST in Gambian children at 4(1/2 months of age who either received BCG vaccination at birth (Group 1 or were BCG naïve (Group 2 in order to examine the role of BCG vaccination and/or exposure to environmental mycobacteria in TST reactivity at this age. Nearly half of the BCG vaccinated children had a positive TST (>or=5 mm whereas all the BCG naïve children were non-reactive, confirming that recent BCG vaccination affects TST reactivity. The BCG naïve children demonstrated in vitro PPD responses in peripheral blood in the absence of TST reactivity, supporting exposure to and priming by environmental mycobacterial antigens. Group 2 were then vaccinated at 4(1/2 months of age and a repeat TST was performed at 20-28 months of age. Positive reactivity (>or=5 mm was evident in 11.1% and 12.5% infants from Group 1 and Group 2 respectively suggesting that the timing of BCG vaccination had little effect by this age. We further assessed for immune correlates in peripheral blood at 4(1/2 months of age. Mycobacterial specific IFNgamma responses were greater in TST responders than in non-responders, although the size of induration did not correlate with IFNgamma. However the IFNgamma: IL-10 ratio positively correlated with TST induration suggesting that the relationship between PPD induced IFNgamma and IL-10 in the peripheral blood may be important in controlling TST reactivity. Collectively these data provide further insights into how the TST is regulated in early life, and how a positive response might be interpreted.

  19. High incidence of severe cyclosporine neurotoxicity in children affected by haemoglobinopaties undergoing myeloablative haematopoietic stem cell transplantation: early diagnosis and prompt intervention ameliorates neurological outcome

    Directory of Open Access Journals (Sweden)

    Minicucci Fabio


    Full Text Available Abstract Background Neurotoxicity is a recognized complication of cyclosporine A (CSA treatment. The incidence of severe CSA-related neurological complications following hematopoietic stem cell transplantation (HSCT is 4-11%. Methods We describe 6 cases of CSA related neurotoxicity out of 67 matched related HSCT performed in paediatric Middle East patients affected by haemoglobinopaties (5 beta thalassemia major, 1 sickle cell disease-SCD. Conditioning regimen consisted of iv busulphan, cyclophosphamide and graft-versus-host-disease (GvHD prophylaxis with CSA, methylprednisolone, methotrexate and ATG. Results All 6 patients presented prodromes such as arterial hypertension, headache, visual disturbances and vomiting, one to two days before overt CSA neurotoxicity. CSA neurotoxicity consisted of generalized seizures, signs of endocranial hypertension and visual disturbances at a median day of onset of 11 days after HSCT (range +1 to +40. Brain magnetic resonance imaging (MRI performed in all subjects showed reversible leukoencephalopathy predominantly in the posterior regions of the brain (PRES in 5/6 patients. EEG performed in 5/6 patients was always abnormal. Neurotoxicity was not explainable by high CSA blood levels, as all patients had CSA in the therapeutic range with a median of 178 ng/ml (range 69-250. CSA was promptly stopped and switched to tacrolimus with disappearance of clinical and radiological findings. All patients are symptoms-free at a median follow up of 882 days (range 60-1065. Conclusions Our experience suggests that paediatric patients with haemoglobinopaties have a high incidence of CSA related neurological events with no correlation between serum CSA levels and neurotoxicity. Prognosis is good following CSA removal. Specific prodromes such as arterial hypertension, headache or visual disturbances occurring in the early post-transplant period should be carefully evaluated with electrophysiological and MRI-based imaging in

  20. Enzymatic Reactions in Microfluidic Devices (United States)

    Ristenpart, W. D.; Wan, J.; Stone, H. A.


    We establish simple scaling laws for enzymatic reactions in microfluidic devices, and we demonstrate that kinetic parameters obtained conventionally using multiple stop-flow experiments may instead be extracted from a single microfluidic experiment. Introduction of an enzyme and substrate species in different arms of a Y-shaped channel allows the two species to diffuse across the parallel streamlines and to begin reacting. Measurements of the product concentration versus distance down the channel provide information about the kinetics of the reaction. In the limit where the enzyme is much larger (and thus less diffusive) than the substrate, we show that near the entrance the total amount of product (P) formed varies as a power law in the distance x down the channel. For reactions that follow standard Michaelis-Menten kinetics, the power law takes the form P˜(Vmax/Km) x^5/2, where Vmax and Km are the maximum reaction rate and Michaelis constant respectively. If a large excess of substrate is used, then Km is identified by measuring Vmax far downstream where the different species are completely mixed by diffusion. Numerical simulations and experiments using the bioluminescent reaction between luciferase and ATP as a model system are both shown to accord with the model. We discuss the implications for significant savings in the amount of time and enzyme required for determination of kinetic parameters.

  1. Bioethanol production: Pretreatment and enzymatic hydrolysis of softwood

    Energy Technology Data Exchange (ETDEWEB)

    Tengborg, Charlotte


    The enzymatic hydrolysis process can be used to produce bioethanol from softwood, which are the dominating raw material in the Northern hemisphere. This thesis deals with the development of the process focusing on the pretreatment and the enzymatic hydrolysis stages. The influence of pretreatment conditions on sugar yield, and the effect of inhibitors on the ethanol yield, were investigated for spruce and pine. The maximum yields of hemicellulose sugars and glucose were obtained under different pretreatment conditions. This indicates that two-stage pretreatment may be preferable. The added catalysts, H{sub 2}SO{sub 4} and SO{sub 2}, resulted in similar total sugar yields about 40 g/100 g dry raw material. However, the fermentability of SO{sub 2}-impregnated material was better. This pretreatment resulted in the formation of inhibitors to the subsequent process steps, e.g. sugar and lignin degradation products. The glucose yield in the enzymatic hydrolysis stage was affected by various parameters such as enzyme loading, temperature, pH, residence time, substrate concentration, and agitation. To decrease the amount of fresh water used and thereby waste water produced, the sugar-rich prehydrolysate from the pretreatment step was included in the enzymatic hydrolysis of the solid fraction, resulting in a reduction in the cellulose conversion of up to 36%. Different prehydrolysate detoxification methods, such as treatment with Ca(OH){sub 2}, laccase, and fermentation using yeast, were investigated. The latter was shown to be very efficient. The amount of fresh water used can be further reduced by recycling various process streams. This was simulated experimentally in a bench-scale process. A reduction in fresh water demand of 50% was obtained without any further negative effects on either hydrolysis or fermentation.

  2. Electrochemical, Chemical and Enzymatic Oxidations of Phenothiazines

    NARCIS (Netherlands)

    Blankert, B.; Hayen, H.; Leeuwen, van S.M.; Karst, U.; Bodoki, E.; Lotrean, S.; Sandulescu, R.; Mora Diaz, N.; Dominguez, O.; Arcos, J.; Kauffmann, J.-M.


    The oxidation of several phenothiazine drugs (phenothiazine, promethazine hydrochloride, promazine hydrochloride, trimeprazine hydrochloride and ethopropazine hydrochloride) has been carried out in aqueous acidic media by electrochemical, chemical and enzymatic methods. The chemical oxidation was pe


    Many immunotoxic compounds, such as benzene and other organic solvents, pesticides, mycotoxins and polycyclic aromatic hydrocarbons, can alter immune function only after undergoing enzyme-mediated reactions within various tissues. In the review that follows, the role of enzymatic...

  4. Small Variations in Early-Life Environment Can Affect Coping Behaviour in Response to Foraging Challenge in the Three-Spined Stickleback.

    Directory of Open Access Journals (Sweden)

    M Rohaa Langenhof

    Full Text Available An increasing concern in the face of human expansion throughout natural habitats is whether animal populations can respond adaptively when confronted with challenges like environmental change and novelty. Behavioural flexibility is an important factor in estimating the adaptive potential of both individuals and populations, and predicting the degree to which they can cope with change.This study on the three-spined stickleback (Gasterosteus aculeatus is an empiric illustration of the degree of behavioural variation that can emerge between semi-natural systems within only a single generation. Wild-caught adult sticklebacks (P, N = 400 were randomly distributed in equal densities over 20 standardized semi-natural environments (ponds, and one year later offspring (F1, N = 652 were presented with repeated behavioural assays. Individuals were challenged to reach a food source through a novel transparent obstacle, during which exploration, activity, foraging, sociability and wall-biting behaviours were recorded through video observation. We found that coping responses of individuals from the first generation to this unfamiliar foraging challenge were related to even relatively small, naturally diversified variation in developmental environment. All measured behaviours were correlated with each other. Especially exploration, sociability and wall-biting were found to differ significantly between ponds. These differences could not be explained by stickleback density or the turbidity of the water.Our findings show that a differences in early-life environment appear to affect stickleback feeding behaviour later in life; b this is the case even when the environmental differences are only small, within natural parameters and diversified gradually; and c effects are present despite semi-natural conditions that fluctuate during the year. Therefore, in behaviourally plastic animals like the stickleback, the adaptive response to human-induced habitat disturbance

  5. Nanostructured Metal Oxides Based Enzymatic Electrochemical Biosensors


    Ansari, Anees A.; Alhoshan, M.; M. S. AlSalhi; Aldwayyan, A.S.


    The unique electrocatalytic properties of the metal oxides and the ease of metal oxide nanostructured fabrication make them extremely interesting materials for electrochemical enzymatic biosensor applications. The application of nanostructured metal oxides in such sensing devices has taken off rapidly and will surely continue to expand. This article provides a review on current research status of electrochemical enzymatic biosensors based on various new types of nanostructured metal oxides su...

  6. Enzymatic biodiesel production: Technical and economical considerations

    DEFF Research Database (Denmark)

    Munk Nielsen, Per; Brask, Jesper; Fjerbæk, Lene


    It is well documented in the literature that enzymatic processing of oils and fats for biodiesel is technically feasible. However, with very few exceptions, enzyme technology is not currently used in commercial-scale biodiesel production. This is mainly due to non-optimized process design and a l...... that enzyme catalysts can become cost effective compared to chemical processing. This work reviews the enzymatic processing of oils and fats into biodiesel with focus on process design and economy....

  7. Rheological Properties of Enzymatically Isolated Tomato Fruit Cuticle. (United States)

    Petracek, P. D.; Bukovac, M. J.


    Rheological properties were determined for cuticular membranes (CMs) enzymatically isolated from mature tomato (Lycopersicon esculentum Mill. cv Pik Red) fruit. The cuticle responded as a viscoelastic polymer in stress-strain studies. Both CM and dewaxed CM expanded and became more elastic and susceptible to fracture when hydrated, suggesting that water plasticized the cuticle. Dewaxing of the CM caused similar changes in elasticity and fracturing, indicating that wax may serve as a supporting filler in the cutin matrix. Exposure of the cuticle to the surfactant Triton X-100 did not significantly affect its rheological properties.

  8. Effect of enzymatic depolymerization on physicochemical and rheological properties of guar gum. (United States)

    Mudgil, Deepak; Barak, Sheweta; Khatkar, B S


    Depolymerization of guar gum using enzymatic hydrolysis was performed to obtain depolymerized guar gum having functional application as soluble dietary fiber. Enzymatic hydrolysis of guar gum significantly affected the physicochemical and rheological characteristics of guar gum. The depolymerized guar gum showed a significant increase in crystallinity index from 3.86% to 13.2% and flow behavior index from 0.31 to 1.7 as compared to native guar gum. Remarkable decrease in intrinsic viscosity and consistency index was also observed from 9 to 0.28 and 4.04 to 0.07, respectively. Results revealed that enzymatic hydrolysis of guar gum resulted in a polysaccharide with low degree of polymerization, viscosity and consistency which could make it useful for incorporation in food products as dietary fiber without affecting the rheology, consistency and texture of the products.

  9. Improved enzymatic hydrolysis of wheat straw by combined use of gamma ray and dilute acid for bioethanol production (United States)

    Hyun Hong, Sung; Taek Lee, Jae; Lee, Sungbeom; Gon Wi, Seung; Ju Cho, Eun; Singh, Sudhir; Sik Lee, Seung; Yeoup Chung, Byung


    Pretreating wheat straw with a combination of dilute acid and gamma irradiation was performed in an attempt to enhance the enzymatic hydrolysis for bioethanol production. The glucose yield was significantly affected by combined pretreatment (3% sulfuric acid-gamma irradiation), compared with untreated wheat straw and individual pretreatment. The increasing enzymatic hydrolysis after combined pretreatment is resulting from decrease in crystallinity of cellulose, loss of hemicelluloses, and removal or modification of lignin. Therefore, combined pretreatment is one of the most effective methods for enhancing the enzymatic hydrolysis of wheat straw biomass.

  10. Structural perspective on enzymatic halogenation. (United States)

    Blasiak, Leah C; Drennan, Catherine L


    Simple halogen substituents frequently afford key structural features that account for the potency and selectivity of natural products, including antibiotics and hormones. For example, when a single chlorine atom on the antibiotic vancomycin is replaced by hydrogen, the resulting antibacterial activity decreases by up to 70% ( Harris , C. M. ; Kannan , R. ; Kopecka , H. ; Harris , T. M. J. Am. Chem. Soc. 1985 , 107 , 6652 - 6658 ). This Account analyzes how structure underlies mechanism in halogenases, the molecular machines designed by nature to incorporate halogens into diverse substrates. Traditional synthetic methods of integrating halogens into complex molecules are often complicated by a lack of specificity and regioselectivity. Nature, however, has developed a variety of elegant mechanisms for halogenating specific substrates with both regio- and stereoselectivity. An improved understanding of the biological routes toward halogenation could lead to the development of novel synthetic methods for the creation of new compounds with enhanced functions. Already, researchers have co-opted a fluorinase from the microorganism Streptomyces cattleya to produce (18)F-labeled molecules for use in positron emission tomography (PET) ( Deng , H. ; Cobb , S. L. ; Gee , A. D. ; Lockhart , A. ; Martarello , L. ; McGlinchey , R. P. ; O'Hagan , D. ; Onega , M. Chem. Commun. 2006 , 652 - 654 ). Therefore, the discovery and characterization of naturally occurring enzymatic halogenation mechanisms has become an active area of research. The catalogue of known halogenating enzymes has expanded from the familiar haloperoxidases to include oxygen-dependent enzymes and fluorinases. Recently, the discovery of a nucleophilic halogenase that catalyzes chlorinations has expanded the repertoire of biological halogenation chemistry ( Dong , C. ; Huang , F. ; Deng , H. ; Schaffrath , C. ; Spencer , J. B. ; O'Hagan , D. ; Naismith , J. H. Nature 2004 , 427 , 561 - 565 ). Structural

  11. An investigation of nitrile transforming enzymes in the chemo-enzymatic synthesis of the taxol sidechain. (United States)

    Wilding, Birgit; Veselá, Alicja B; Perry, Justin J B; Black, Gary W; Zhang, Meng; Martínková, Ludmila; Klempier, Norbert


    Paclitaxel (taxol) is an antimicrotubule agent widely used in the treatment of cancer. Taxol is prepared in a semisynthetic route by coupling the N-benzoyl-(2R,3S)-3-phenylisoserine sidechain to the baccatin III core structure. Precursors of the taxol sidechain have previously been prepared in chemoenzymatic approaches using acylases, lipases, and reductases, mostly featuring the enantioselective, enzymatic step early in the reaction pathway. Here, nitrile hydrolysing enzymes, namely nitrile hydratases and nitrilases, are investigated for the enzymatic hydrolysis of two different sidechain precursors. Both sidechain precursors, an openchain α-hydroxy-β-amino nitrile and a cyanodihydrooxazole, are suitable for coupling to baccatin III directly after the enzymatic step. An extensive set of nitrilases and nitrile hydratases was screened towards their activity and selectivity in the hydrolysis of two taxol sidechain precursors and their epimers. A number of nitrilases and nitrile hydratases converted both sidechain precursors and their epimers.

  12. Early Paleogene Arctic terrestrial ecosystems affected by the change of polar hydrology under global warming:Implications for modern climate change at high latitudes

    Institute of Scientific and Technical Information of China (English)

    Gaytha; A.; LANGLOIS


    Our understanding of both the role and impact of Arctic environmental changes under the current global warming climate is rather limited despite efforts of improved monitoring and wider assessment through remote sensing technology. Changes of Arctic ecosystems under early Paleogene warming climate provide an analogue to evaluate long-term responses of Arctic environmental alteration to global warming. This study reviews Arctic terrestrial ecosystems and their transformation under marked change of hydrological conditions during the warmest period in early Cenozoic, the Paleocene and Eocene. We describe a new approach to quantitatively reconstruct high latitudinal paleohydrology using compound-specific hydrogen isotope analysis which applies empirically derived genus-specific hydrogen isotope fractionations to in situ biomolecules from fossil plants. We propose a moisture recycling model at the Arctic to explain the reconstructed hydrogen isotope signals of ancient high latitude precipitation during early Paleogene, which bears implications to the likely change of modern Arctic ecosystems under the projected accelerated global warming.

  13. Impact of the redox-cycling herbicide diquat on transcript expression and antioxidant enzymatic activities of the freshwater snail Lymnaea stagnalis

    Energy Technology Data Exchange (ETDEWEB)

    Bouetard, Anthony, E-mail: [INRA, UMR INRA-Agrocampus Ouest ESE 0985, Equipe Ecotoxicologie et Qualite des Milieux Aquatiques, 65 rue de Saint-Brieuc, 35042 Rennes cedex (France); Besnard, Anne-Laure; Vassaux, Daniele; Lagadic, Laurent; Coutellec, Marie-Agnes [INRA, UMR INRA-Agrocampus Ouest ESE 0985, Equipe Ecotoxicologie et Qualite des Milieux Aquatiques, 65 rue de Saint-Brieuc, 35042 Rennes cedex (France)


    significantly (or non-significantly for cat) after 5 h of exposure, and went back to control levels afterwards, suggesting the onset of an early response to oxidative stress associated to the unbalance of reactive oxygen species (ROS) in hepatocytes. Although increases obtained for Gred and SOD activities were globally consistent with their respective transcript expressions, up-regulation of transcription was not always correlated with increase of enzymatic activity, indicating that diquat might affect steps downstream of transcription. However, constitutive levels of enzymatic activities were at least maintained. In conclusion, diquat was shown to affect expression of the whole set of studied transcripts, reflecting their suitability as markers of early response to oxidative stress in L. stagnalis.

  14. Effects of Ionic Strength on the Enzymatic Hydrolysis of Diluted and Concentrated Whey Protein Isolate

    NARCIS (Netherlands)

    Butré, C.I.; Wierenga, P.A.; Gruppen, H.


    To identify the parameters that affect enzymatic hydrolysis at high substrate concentrations, whey protein isolate (1–30% w/v) was hydrolyzed by Alcalase and Neutrase at constant enzyme-to-substrate ratio. No changes were observed in the solubility and the aggregation state of the proteins. With inc

  15. Early Onset Inflammation in Pre-Insulin-Resistant Diet-Induced Obese Rats Does Not Affect the Vasoreactivity of Isolated Small Mesenteric Arteries

    DEFF Research Database (Denmark)

    Blædel, Martin; Raun, Kirsten; Boonen, Harrie C M;


    Background: Obesity is an increasing burden affecting developed and emerging societies since it is associated with an increased risk of diabetes and consequent cardiovascular complications. Increasing evidence points towards a pivotal role of inflammation in the etiology of vascular dysfunction...

  16. "We Don't Allow Children to Climb Trees": How a Focus on Safety Affects Norwegian Children's Play in Early-Childhood Education and Care Settings (United States)

    Sandseter, Ellen Beate Hansen; Sando, Ole Johan


    The authors point out a basic contradiction: On one hand, we want to keep children as safe as possible; On the other, they suggest, learning to take risks is a normal part of childhood and child development. In Norway, research has shown that early-childhood education and care (ECEC) practitioners have, in the past, taken a permissive approach to…

  17. Does an Early and Aggressive Combined Wrapping and Dangling Procedure Affect the Clinical Outcome of Lower Extremity Free Flaps?-A Randomized Controlled Prospective Study Using Microdialysis Monitoring. (United States)

    Neubert, Nils; Vogt, P M; May, M; Boyce, M; Koenneker, S; Budde, E; Jokuszies, A


    Background The ideal starting point for flap training (FT) with the combined wrapping and dangling procedure is still a question of debate. Most units follow their own established protocols and currently evidence of flap compromise due to FT is still lacking. The aim of this study was to prove if an early and "aggressive" wrapping and dangling protocol could lead to metabolic changes, measured by microdialysis, indicating ischemia resulting in compromised flap perfusion. Methods Between 2010 and 2014, 49 patients with microvascular free flap reconstruction of the lower limb were included in this study. Patients were randomized into two groups. Group I started FT on the 7th postoperative day, and group II started on day 3. FT consisted of a combined wrapping and dangling procedure doubling its duration daily and ending at day 5. Flaps were monitored clinically and by microdialysis for ischemia-induced changes and metabolic parameters in the flap tissue in respect to different starting points of FT. Results All 49 patients in both groups were able to complete the postoperative FT without complications. Noninferiority of the early group could be proven and microdialysis results showed no differences between both groups. Conclusion We could prove by microdialysis that an early start of FT does not lead to compromised flap perfusion. Moreover, an early start of FT can lead to a reduced length of hospital stay. Furthermore, a reduced risk for deep vein thrombosis, pulmonary embolism, and pneumonia due to earlier mobilization might be an appreciated side effect.

  18. Enzymatic treatment of estrogens and estrogen glucuronide

    Institute of Scientific and Technical Information of China (English)

    Takaaki Tanaka; Toshiyuki Tamura; Yuuichi Ishizaki; Akito Kawasaki; Tomokazu Kawase; Masahiro Teraguchi; Masayuki Taniguchi


    Natural and synthetic estrogens from sewage treatment systems are suspected to influence the reproductive health of the animals in the rivers.In this article we investigated the enzymatic treatment of three estrogens (estrone,17β-estradiol,and 17α-ethynyletstradiol) by a fungal laccase which oxidize phenolic compounds with dissolved oxygen.The elimination of the estrogenic activities by enzymatic oxidation was demonstrated by medaka vitellogenin assay.In addition,we developed an enzymatic treatment system comprised of β-D-glucuronidase and the laccase for 17β-estradiol 3-(β-D-glucuronide) degradation.The two enzymes eliminated 17β-estradiol 3-(β-D-glucuronide) and the intermediate,17β-estradiol,efficiently.

  19. Effect of macromolecular crowding on the rate of diffusion-limited enzymatic reaction

    Indian Academy of Sciences (India)

    Manish Agrawal; S B Santra; Rajat Anand; Rajaram Swaminathan


    The cytoplasm of a living cell is crowded with several macromolecules of different shapes and sizes. Molecular diffusion in such a medium becomes anomalous due to the presence of macromolecules and diffusivity is expected to decrease with increase in macromolecular crowding. Moreover, many cellular processes are dependent on molecular diffusion in the cell cytosol. The enzymatic reaction rate has been shown to be affected by the presence of such macromolecules. A simple numerical model is proposed here based on percolation and diffusion in disordered systems to study the effect of macromolecular crowding on the enzymatic reaction rates. The model qualitatively explains some of the experimental observations.

  20. Enzymatic production of polysaccharides from gum tragacanth

    DEFF Research Database (Denmark)


    Plant polysaccharides, relating to the field of natural probiotic components, can comprise structures similar to human milk oligosaccharides. A method for enzymatic hydrolysis of gum tragacanth from the bush-like legumes of the genus Astragalus, using a combination of pectin hydrolases and a xylo......Plant polysaccharides, relating to the field of natural probiotic components, can comprise structures similar to human milk oligosaccharides. A method for enzymatic hydrolysis of gum tragacanth from the bush-like legumes of the genus Astragalus, using a combination of pectin hydrolases...... and a xylogalacturonan hydrolase, is described. Fractions with different oligo- and/or polysaccharide compositions and structure are separated according to molecular weight....

  1. Enzymatic and molecular strategies to diagnose Pompe disease. (United States)

    Reuser, Ajj; Verheijen, Fw; Kroos, Ma; Okumiya, T; Van Diggelen, Op; Van der Ploeg, At; Halley, Djj


    Enzyme replacement therapy for Pompe disease, a neuromuscular disorder characterized by lysosomal glycogen storage due to acid α-glucosidase deficiency, has entered the clinic. There is more than ever a need for early and reliable diagnosis. The objective of this review is to present a critical review of the recent literature on laboratory procedures to diagnose Pompe disease by enzymatic assay and DNA analysis. The methods we used were Compilation and expert interpretation of recent and relevant publications. The introduction of new and the updating of existing laboratory procedures have facilitated the diagnosis of Pompe disease (glycogen storage disease type II; acid maltase deficiency; OMIM 232300). With regard to enzymatic analysis, the application of acarbose as inhibitor of maltase-glucoamylase has enabled the use of mixed leukocyte preparations as diagnostic material. The use of glycogen as a natural substrate in the reaction mixture adds to the selectivity of this procedure. Newborn screening is envisaged and facilitated by the introduction of high-throughput procedures. DNA analysis has become an integral part of the diagnostic procedure for confirmation and completion, for carrier detection, and for genetic counseling.

  2. [Affective dependency]. (United States)

    Scantamburlo, G; Pitchot, W; Ansseau, M


    Affective dependency is characterized by emotional distress (insecure attachment) and dependency to another person with a low self-esteem and reassurance need. The paper proposes a reflection on the definition of emotional dependency and the confusion caused by various denominations. Overprotective and authoritarian parenting, cultural and socio-environmental factors may contribute to the development of dependent personality. Psychological epigenetic factors, such as early socio-emotional trauma could on neuronal circuits in prefronto-limbic regions that are essential for emotional behaviour.We also focus on the interrelations between dependent personality, domestic violence and addictions. The objective for the clinician is to propose a restoration of self-esteem and therapeutic strategies focused on autonomy.

  3. Early brain loss in circuits affected by Alzheimer’s disease is predicted by fornix microstructure but may be independent of gray matter

    Directory of Open Access Journals (Sweden)

    Evan eFletcher


    Full Text Available In a cohort of community-recruited elderly subjects with normal cognition at initial evaluation, we found that baseline fornix white matter microstructure was significantly correlated with early volumetric longitudinal tissue change across a region of interest (called fSROI, which overlaps circuits known to be selectively vulnerable to AD pathology. Other white matter and gray matter regions had much weaker or non-existent associations with longitudinal tissue change. Tissue loss in fSROI was in turn a significant factor in a survival model of cognitive decline, as was baseline fornix microstructure. These findings suggest that WM deterioration in the fornix and tissue loss in fSROI may be the early beginnings of posterior limbic circuit and default mode network degeneration. We also found that gray matter baseline volumes in the entorhinal cortex and hippocampus predicted cognitive decline in survival models. But since GM regions did not also significantly predict brain tissue loss, our results may imply a view in which early, prodromal deterioration appears as two quasi independent processes in white and gray matter regions of the limbic circuit crucial to memory.

  4. Effect of Steam Explosion Pretreatment on Bamboo for Enzymatic Hydrolysis and Ethanol Fermentation

    Directory of Open Access Journals (Sweden)

    Zhiqiang Li


    Full Text Available Based on the steam explosion pretreatment that has been applied to other types of lignocellulosic biomass, the steam explosion pretreatment of bamboo, along with a study of the chemical compositions and enzymatic hydrolyzability of substrates, was conducted. The results show that steam explosion pretreatment can greatly enhance the cellulose-to-glucose conversion yield after enzymatic hydrolysis, which is sometimes affected by bamboo age and steam explosion conditions. When the steam explosion pretreatment conditions were 2.0 MPa (pressure and 4 min (time, the cellulose-to-glucose conversion yield of 2-year-old bamboo substrate was 62.5%. However, the cellulose-to-glucose conversion yield of bamboo substrates after direct (without steam explosion pretreatment sodium chlorite/acetic acid delignification was 93.1%. Fermentation of enzymatic hydrolyzates with Saccharomyces cerevisiae resulted in about 88.1% to 96.2% of the corresponding theoretical ethanol yield after 24 h.

  5. A Miniature Membrane Reactor for Evaluation of Process Design Options on the Enzymatic Degradation of Pectin

    DEFF Research Database (Denmark)

    Zainal Alam, Muhd Nazrul Hisham; Pinelo, Manuel; Arnous, Anis


    The objective of this paper is to assess if a membrane microbioreactor system could potentially be used to diagnose consequences of different process design and reactor operation options relevant for larger-scale enzymatic degradation of pectin reactions. The membrane microbioreactor prototype...... was demonstrated by performing a continuous enzymatic degradation of pectin experiment for a range of reactor conditions: different membrane molecular weight cutoff (MWCO) values, enzyme-to-substrate ratios (E/S), and substrate feeding rates (F) were assessed. Based on the experimental data, it was found...... design affected the membrane rejection profile. The results obtained thus underlined the suitability of a miniature membrane reactor system for evaluating different process design options that are relevant for larger-scale reactions of enzymatic pectin degradation....

  6. Enzymatic production of human milk oligosaccharides

    DEFF Research Database (Denmark)

    Guo, Yao

    Enzymatic treatment of biomass is an environmentally friendly method to obtain a range of value- added products, such as biofuels, animal feed or food ingredients. The objective of this PhD study was to biocatalytically produce biofunctional food ingredients – human milk oligosaccharides decorated...

  7. Saccharide modified silica particles by enzymatic grafting

    NARCIS (Netherlands)

    Loos, Katja; Braunmühl, Volker von; Stadler, Reimund; Landfester, K; Spiess, HW


    The surface of silica particles has been chemically modified with oligo- or poly-(α,1→4)-D-glucopyranose (amylose) chains of various length by covalently attaching maltoheptaose derivatives to the solid support and enzymatic polymerization of glucose-1-phosphate with a potato phosphorylase as cataly

  8. Tandem and sequential multi-enzymatic syntheses

    NARCIS (Netherlands)

    Kim, B.G.; Ahn, J.H.; Sello, G.; Di Gennaro, P.; van Herk, T.; Hartog, A.F.; Wever, R.; Oroz-Guinea, I.; Sánchez-Moreno, I.; García-Junceda, E.; Wu, B.; Szymanski, W.; Feringa, B.L.; Janssen, D.B.; Villo, L.; Kreen, M.; Kudryashova, M.; Metsala, A.; Tamp, S.; Lille, ü.; Pehk, T.; Parve, O.; McClean, K.; Eddowes, P.; Whittall, J.; Sutton, P.W.


    This chapter contains sections titled: Production of Isorhamnetin 3-O-Glucoside in Escherichia coli Using Engineered Glycosyltransferase Multienzymatic Preparation of (−)-3-(Oxiran-2-yl)Benzoic Acid Enzymatic Synthesis of Carbohydrates from Dihydroxyacetone and Aldehydes by a One Pot Enzyme Cascade

  9. Starch facilitates enzymatic wheat gluten hydrolysis

    NARCIS (Netherlands)

    Hardt, N.A.; Boom, R.M.; Goot, van der A.J.


    Wheat gluten can be hydrolyzed by either using (vital) wheat gluten or directly from wheat flour. This study investigates the influence of the presence of starch, the main component of wheat, on enzymatic wheat gluten hydrolysis. Wheat gluten present in wheat flour (WFG) and vital wheat gluten (VWG)

  10. Improved enzymatic production of phenolated glycerides through alkyl phenolate intermediate

    DEFF Research Database (Denmark)

    Yang, Zhiyong; Feddern, Vivian; Glasius, Marianne


    This work reported a novel approach for synthesis of dihydrocaffoylated glycerides, consisting of 2 steps: enzymatic synthesis of octyl dihydrocaffeate (as a synthetic intermediate) from octanol and dihydrocaffeic acid (DHCA), and enzymatic interesterification of triglycerides with octyl dihydroc...

  11. Enzymatic biotransformation of terpenes as bioactive agents. (United States)

    Sultana, Nighat; Saify, Zafar Saeed


    The plant-derived terpenoids are considered to be the most potent anticancer, anti-inflammatory and anticarcinogenic compounds known. Enzymatic biotransformation is a very useful approach to expand the chemical diversity of natural products. Recent enzymatic biotransformation studies on terpenoids have resulted in the isolation of novel compounds. 14-hydroxy methyl caryophyllene oxide produced from caryophyllene oxide showed a potent inhibitory activity against the butyryl cholinesterase enzyme, and was found to be more potent than parent caryophyllene oxide. The metabolites 3β,7β-dihydroxy-11-oxo-olean-12-en-30-oic acid, betulin, betulonic acid, argentatin A, incanilin, 18β glycyrrhetinic acid, 3,11-dioxo-olean-12-en-30-oic acid produced from 18β glycyrrhetinic acid were screened against the enzyme lipoxygenase. 3,11-Dioxo-olean-12-en-30-oic acid, was found to be more active than the parent compound. The metabolites 3β-hydroxy sclareol 18α-hydroxy sclareol, 6α,18α-dihydroxy sclareol, 11S,18α-dihydroxy sclareol, and 1β-hydroxy sclareol and 11S,18α-dihydroxy sclareol produced from sclareol were screened for antibacterial activity. 1β-Hydroxy sclareol was found to be more active than parent sclareol. There are several reports on natural product enzymatic biotransformation, but few have been conducted on terpenes. This review summarizes the classification, advantages and agents of enzymatic transformation and examines the potential role of new enzymatically transformed terpenoids and their derivatives in the chemoprevention and treatment of other diseases.

  12. Delay in liver bud development during early embryogenesis leads to violation of epicardium formation and its epithelial-to-mesenchymal transformation, but do not affect coronary endothelium formation

    Directory of Open Access Journals (Sweden)

    Pototskaya O. Yu.


    Full Text Available Despite intensive investigation of heart embryogenesis the origin of coronary endothelium is still under debate. Existence of close interrelation between proepicardium, liver bud, sinus venosus and early coronary vessels is obvious, but the nature of this interconnection is unclear as well as exact source of endothelial cells. Thus, the purpose of our research was to investigate the effect of inhibition of liver bud development on formation of coronary vessels. To inhibit the liver bud we injected aminoguanidine sulfate into the yolk sack of chick embryos on 14 stage of development by HH (first group - to prevent contact between proepicardium and liver, and on 16 stage (second group to allow contact between proepicardium and liver during a short period of time. In the first group it was observed the violation of epicardium formation and early death during first 3 days of incubation. At embryos it was revealed thinning of myocardium and atrio-ventricular cushions, abnormal looping – presumably due to the absence of epicardium. The death was caused by heart insufficiency. In the second group it was observed the beginning of epicardium formation, but the heart defects were the same as in the first group – presumably due to the absence of epicardium-derived cells. At embryos of this group, who survived till 26 stage of development (4.5 day of incubation, despite absence of cellular component in subepicardial space and almost completely absence of liver, we observed formation of coronary vessels. They were concentrated on the dorsal surface of atrio-vetricular channel and were presented by continuation of endothelium of the sinus venosus.

  13. Oxidative enzymatic gelation of sugar beet pectin for emulsion stabilization

    DEFF Research Database (Denmark)

    Abang Zaidel, Dayang Norulfairuz; Meyer, Anne S.


    Pectin from sugar beet is derived from the sugar beet pulp residue which results when sugar beets are processed for sucrose extraction. The sugar beet pectin has poor gelationability by the classic divalentcation molecular mechanism because of a relatively high acetylation degree and short...... polygalacturonate backbone chain length. However, due to the feruloyl-substitutions on the side chains, the sugar beet pectic polysaccharides can be cross-linked via enzyme catalyzed oxidation. The enzyme kinetics and functionality of such oxidativelycross-linked sugar beet pectin, in relation to stabilizing...... emulsions has recently been investigated in model food emulsions. This paper reviews the pectin chemistry, enzymatic oxidative gelation mechanisms, interaction mechanisms of the sugar beet pectin with the emulsion droplets and explores how the gelation affects the rheology and stability of emulsion systems...

  14. Enzymatic Modification of Antioxidants Towards Omega-3 Oil Protection

    DEFF Research Database (Denmark)

    Yang, Zhiyong

    This PhD dissertation entitled “Enzymatic Modification of Antioxidants Towards Omega-3 Oil Protection” is primarily focused on synthesize of novel antioxidant from natural sources for better protection of oxidation-prone omega 3 oil. Selected phenolic acids were conjugated with fatty alcohols...... in different chain length and triacylglycerol (TAG). Several synthesis strategies were evaluated. Synthesis of lipophilic phenolic fatty alcohols esters were initially conducted in a binary organic solvent system, which was composed of hexane and butanone. Preliminary studies were conducted to identify...... the main parameters affecting yield of these novel lipophilized phenolic fatty alcohol esters in a binary solvent system. The parameters studied in these preliminary studies included volume ratio of hexane to butanone, chain lengths of fatty alcohols and types of phenolic acids (dihydrocaffeic acid (DHCA...

  15. Early-age feed restriction affects viability and gene expression of satellite cells isolated from the gastrocnemius muscle of broiler chicks

    Directory of Open Access Journals (Sweden)

    Li Yue


    Full Text Available Abstract Background Muscle growth depends on the fusion of proliferate satellite cells to existing myofibers. We reported previously that 0–14 day intermittent feeding led to persistent retardation in myofiber hypertrophy. However, how satellite cells respond to such nutritional insult has not been adequately elucidated. Results One-day-old broiler chicks were allocated to control (Con, ad libitum feeding, intermittent feeding (IF, feed provided on alternate days and re-feeding (RF, 2 days ad libitum feeding after 12 days of intermittent feeding groups. Chickens were killed on Day 15 and satellite cells were isolated. When cultured, satellite cells from the IF group demonstrated significant retardation in proliferation and differentiation potential, while RF partly restored the proliferation rate and differentiation potential of the satellite cells. Significant up-regulation of insulin like growth factor I receptor (IGF-IR (P0.05 and thyroid hormone receptor α (TRα (P0.05, and down-regulation of growth hormone receptor (GHR (P0.01 and IGF-I (P0.01 mRNA expression was observed in freshly isolated IF satellite cells when compared with Con cells. In RF cells, the mRNA expression of IGF-I was higher (P0.05 and of TRα was lower (P0.01 than in IF cells, suggesting that RF restored the mRNA expression of TRα and IGF-I, but not of GHR and IGF-IR. The Bax/Bcl-2 ratio tended to increase in the IF group, which was reversed in the RF group (P0.05, indicating that RF reduced the pro-apoptotic influence of IF. Moreover, no significant effect of T3 was detected on cell survival in IF cells compared with Con (PP0.05 cells. Conclusions These data suggest that early-age feed restriction inhibits the proliferation and differentiation of satellite cells, induces changes in mRNA expression of the GH/IGF-I and thyroid hormone receptors in satellite cells, as well as blunted sensitivity of satellite cells to T3, and that RF partially reverses these effects. Thus

  16. Enzymatic hydrolysis of poly(ethylene furanoate). (United States)

    Pellis, Alessandro; Haernvall, Karolina; Pichler, Christian M; Ghazaryan, Gagik; Breinbauer, Rolf; Guebitz, Georg M


    The urgency of producing new environmentally-friendly polyesters strongly enhanced the development of bio-based poly(ethylene furanoate) (PEF) as an alternative to plastics like poly(ethylene terephthalate) (PET) for applications that include food packaging, personal and home care containers and thermoforming equipment. In this study, PEF powders of various molecular weights (6, 10 and 40kDa) were synthetized and their susceptibility to enzymatic hydrolysis was investigated for the first time. According to LC/TOF-MS analysis, cutinase 1 from Thermobifida cellulosilytica liberated both 2,5-furandicarboxylic acid and oligomers of up to DP4. The enzyme preferentially hydrolyzed PEF with higher molecular weights but was active on all tested substrates. Mild enzymatic hydrolysis of PEF has a potential both for surface functionalization and monomers recycling.

  17. Production of MAG via enzymatic glycerolysis (United States)

    Jamlus, Norul Naziraa Ahmad; Derawi, Darfizzi; Salimon, Jumat


    Enzymatic glycerolysis of a medium chain methyl ester, methyl laurate was performed using lipase Candida antarctica (Novozyme 435) for 6 hours at 55°C. The percentage of components mixture of product were determined by using gas chromatography technique. The enzymatic reaction was successfully produced monolaurin (45.9 %), dilaurin (47.1 %) and trilaurin (7.0 %) respectively. Thin layer chromatography (TLC) plate also showed a good separation of component spots. Fourier transformation infra-red (FTIR) spectrum showed the presence of ester carbonyl at wavenumber 1739.99 cm-1 and hydrogen bonded O-H at 3512.03 cm-1. The product is potentially to be used as emulsifier and additive in food industry, pharmaceutical, as well as antibacterial.

  18. Enzymatic hydrolysis of corn bran arabinoxylan

    DEFF Research Database (Denmark)

    Agger, Jane

    This thesis concerns enzymatic hydrolysis of corn bran arabinoxylan. The work has focused on understanding the composition and structure of corn bran with specific interest in arabinoxylan with the main purpose of targeting enzymatic hydrolysis for increased yields. Corn bran has been used...... as a model substrate because it represents a readily available agroindustrial side product with upgrading potentials. Corn bran originates from the wet-milling process in corn starch processing, is the outmost layers of the corn kernel and is particularly rich in pentose monosaccharides comprising the major...... components of arabinoxylan. Corn bran is one of the most recalcitrant cereal byproducts with arabinoxylans of particular heterogeneous nature. It is also rich in feruloyl derived substitutions, which are responsible for extensive cross-linking between arabinoxylan molecules and thereby participate...

  19. Enzymatic Aqueous Extraction of Soybean Oil

    Institute of Scientific and Technical Information of China (English)


    The procedure of enzymatic aqueous extraction of soybean oil was assessed when two-step controlled enzymatic hydrolysis was applied. With aqueous extraction of soybean oil-containing protein, the highest yield of oil was 96.1% at the optimized conditions studied. Soybean oil-containing protein was hydrolyzed and resulted in releasing part of oil. The separated protein that contained 40% oil was enriched due to its adsorption capacity of released oil, the average oil extraction yeild reached 93.5%. Then the high oil content protein was hydrolyzed again to release oil by enzyme, the oil extraction yeild was 80.4%. As a result, high quality of soybean oil was obtained and the content of total oil yield was 74.4%.

  20. Operation and Control of Enzymatic Biodiesel Production

    DEFF Research Database (Denmark)

    Price, Jason Anthony; Huusom, Jakob Kjøbsted; Nordblad, Mathias

    -product. Current literature indicates that enzymatic processing of oils and fats to produce biodiesel is technically feasible and developments in immobilization technology indicate that enzyme catalysts can become cost effective compared to chemical processing. However, with very few exceptions, enzyme technology...... is not currently used in commercial-scale biodiesel production. This is mainly due to non-optimized process designs, which do not use the full potential of the catalysts in a cost-efficient way. Furthermore is it unclear what process variables need to be monitored and controlled to ensure optimal economics...... an enzymatic route, batch operation is a straightforward and efficient means for producing BD with its main disadvantage being the downtime between batches. For large-scale production of biodiesel, continuous operation is an attractive alternative as it enables efficient use of manpower and capital assets...

  1. A Networks Approach to Modeling Enzymatic Reactions. (United States)

    Imhof, P


    Modeling enzymatic reactions is a demanding task due to the complexity of the system, the many degrees of freedom involved and the complex, chemical, and conformational transitions associated with the reaction. Consequently, enzymatic reactions are not determined by precisely one reaction pathway. Hence, it is beneficial to obtain a comprehensive picture of possible reaction paths and competing mechanisms. By combining individually generated intermediate states and chemical transition steps a network of such pathways can be constructed. Transition networks are a discretized representation of a potential energy landscape consisting of a multitude of reaction pathways connecting the end states of the reaction. The graph structure of the network allows an easy identification of the energetically most favorable pathways as well as a number of alternative routes.

  2. Age-related deficits of conflict resolution mainly affect early information processing stages – an ERP study with different conflict types

    Directory of Open Access Journals (Sweden)

    Margarethe eKorsch


    Full Text Available Aging is usually accompanied by alterations of cognitive control functions such as conflict processing. Recent research suggests that aging effects on cognitive control seem to vary with degree and source of conflict, and conflict specific aging effects on performance measures as well as neural activation patterns have been shown. However, there is sparse information whether and how aging affects different stages of conflict processing as indicated by event related potentials (ERP such as the P2, N2 and P3 components. In the present study 19 young and 23 elderly adults performed a combined Flanker conflict and stimulus-response-conflict (SRC task. Analysis of the median reaction times revealed an increased SRC related conflict effect in elderly. ERP analysis furthermore demonstrated an age-related increase of the P2 amplitude in response to the SRC task. In addition, elderly adults exhibited an increased P3 amplitude modulation induced by incongruent SRC and Flanker conflict trials.

  3. Enzymatic Hydrolysis of Oleuropein from Olea europea (Olive Leaf Extract and Antioxidant Activities

    Directory of Open Access Journals (Sweden)

    Jiao-Jiao Yuan


    Full Text Available Oleuropein (OE, the main polyphenol in olive leaf extract, is likely to decompose into hydroxytyrosol (HT and elenolic acid under the action of light, acid, base, high temperature. In the enzymatic process, the content of OE in olive leaf extract and enzyme are key factors that affect the yield of HT. A selective enzyme was screened from among 10 enzymes with a high OE degradation rate. A single factor (pH, temperature, time, enzyme quantity optimization process and a Box-Behnken design were studied for the enzymatic hydrolysis of 81.04% OE olive leaf extract. Additionally, enzymatic hydrolysis results with different substrates (38.6% and 81.04% OE were compared and the DPPH antioxidant properties were also evaluated. The result showed that the performance of hydrolysis treatments was best using hemicellulase as a bio-catalyst, and the high purity of OE in olive extract was beneficial to biotransform OE into HT. The optimal enzymatic conditions for achieving a maximal yield of HT content obtained by the regression were as follows: pH 5, temperature 55 °C and enzyme quantity 55 mg. The experimental result was 11.31% ± 0.15%, and the degradation rate of OE was 98.54%. From the present investigation of the antioxidant activity determined by the DPPH method, the phenol content and radical scavenging effect were both decreased after enzymatic hydrolysis by hemicellulase. However, a high antioxidant activity of the ethyl acetate extract enzymatic hydrolysate (IC50 = 41.82 μg/mL was demonstated. The results presented in this work suggested that hemicellulase has promising and attractive properties for industrial production of HT, and indicated that HT might be a valuable biological component for use in pharmaceutical products and functional foods.

  4. Early high-sodium solid diet does not affect sodium intake, sodium preference, blood volume and blood pressure in adult Wistar-Kyoto rats. (United States)

    Ufnal, Marcin; Drapala, Adrian; Sikora, Mariusz; Zera, Tymoteusz


    A high-Na diet may lead to the development of hypertension in both humans and rats; however, the causes of Na intake in amounts greater than physiologically needed as well as the mechanisms whereby high-Na food elevates blood pressure are not clear. Therefore, we decided to test the hypothesis that a high-Na diet introduced after suckling affects Na intake, food preference, resting blood pressure and blood volume in adult rats. Male Wistar-Kyoto (WKY) rats, 4 weeks old, were divided into three groups and placed on either a high-Na (3.28%), a medium-Na (0.82%) or a regular diet (0.22%) with the same energy content for 8 weeks. Subsequently, food preference, resting arterial blood pressure, blood volume, plasma osmolality and Na blood level were evaluated. When offered a choice of diets, all the groups preferred the regular chow, and there was no significant difference in total Na intake between the groups. When the rats experienced the change from their initial chow to a new one with different Na content, they continued to eat the same amount of food. Body weight, resting arterial blood pressure, blood volume, plasma osmolality and Na blood level were comparable between the groups. In conclusion, the results show that a high-Na diet introduced immediately after suckling does not affect Na preference and Na intake in adult WKY rats. Furthermore, the findings provide evidence that both blood volume and arterial blood pressure are highly protected in normotensive rats on a high-Na diet.

  5. High Intensity Interval Training Favourably Affects Angiotensinogen mRNA Expression and Markers of Cardiorenal Health in a Rat Model of Early-Stage Chronic Kidney Disease

    Directory of Open Access Journals (Sweden)

    Patrick S. Tucker


    Full Text Available The majority of CKD-related complications stem from cardiovascular pathologies such as hypertension. To help reduce cardiovascular complications, aerobic exercise is often prescribed. Emerging evidence suggests high intensity interval training (HIIT may be more beneficial than traditional aerobic exercise. However, appraisals of varying forms of aerobic exercise, along with descriptions of mechanisms responsible for health-related improvements, are lacking. This study examined the effects of 8 weeks of HIIT (85% VO2max, versus low intensity aerobic exercise (LIT; 45–50% VO2max and sedentary behaviour (SED, in an animal model of early-stage CKD. Tissue-specific mRNA expression of RAAS-related genes and CKD-related clinical markers were examined. Compared to SED, HIIT resulted in increased plasma albumin (p=0.001, reduced remnant kidney weight (p=0.028, and reduced kidney weight-body weight ratios (p=0.045. Compared to LIT, HIIT resulted in reduced Agt mRNA expression (p=0.035, reduced plasma LDL (p=0.001, triglycerides (p=0.029, and total cholesterol (p=0.002, increased plasma albumin (p=0.047, reduced remnant kidney weight (p=0.005, and reduced kidney weight-body weight ratios (p=0.048. These results suggest HIIT is a more potent regulator of several markers that describe and influence health in CKD.

  6. Tropheryma whipplei, the agent of Whipple's disease, affects the early to late phagosome transition and survives in a Rab5- and Rab7-positive compartment.

    Directory of Open Access Journals (Sweden)

    Giovanna Mottola

    Full Text Available Tropheryma whipplei, the agent of Whipple's disease, inhibits phago-lysosome biogenesis to create a suitable niche for its survival and replication in macrophages. To understand the mechanism by which it subverts phagosome maturation, we used biochemical and cell biological approaches to purify and characterise the intracellular compartment where Tropheryma whipplei resides using mouse bone-marrow-derived macrophages. We showed that in addition to Lamp-1, the Tropheryma whipplei phagosome is positive for Rab5 and Rab7, two GTPases required for the early to late phagosome transition. Unlike other pathogens, inhibition of PI(3P production was not the mechanism for Rab5 stabilisation at the phagosome. Overexpression of the inactive, GDP-bound form of Rab5 bypassed the pathogen-induced blockade of phago-lysosome biogenesis. This suggests that Tropheryma whipplei blocks the switch from Rab5 to Rab7 by acting on the Rab5 GTPase cycle. A bio-informatic analysis of the Tropheryma whipplei genome revealed a glyceraldehyde-3-phosphate dehydrogenase (GAPDH homologous with the GAPDH of Listeria monocytogenes, and this may be the bacterial protein responsible for blocking Rab5 activity. To our knowledge, Tropheryma whipplei is the first pathogen described to induce a "chimeric" phagosome stably expressing both Rab5 and Rab7, suggesting a novel and specific mechanism for subverting phagosome maturation.

  7. Agrobacterium-mediated transformation of apple (Malus x domestica Borkh.): an assessment of factors affecting gene transfer efficiency during early transformation steps. (United States)

    De Bondt, A; Eggermont, K; Druart, P; De Vil, M; Goderis, I; Vanderleyden, J; Broekaert, W F


    The factors influencing transfer of an intron - containing β-glucuronidase gene to apple leaf explants were studied during early steps of an Agrobacterium tumefaciens-mediated transformation procedure. The gene transfer process was evaluated by counting the number of β-glucuronidase expressing leaf zones immediately after cocultivation, as well as by counting the number of β-glucuronidase expressing calli developing on the explants after 6 weeks of postcultivation in the presence of 50 mg/l kanamycin. Of three different tested disarmed A. tumefaciens strains, EHA101(pEHA101) was the most effective for apple transformation. Cocultivation of leaf explants with A. tumefaciens on a medium with a high cytokinin level was more conducive to gene transfer than cocultivation on media with high auxin concentrations. Precultivation of leaf explants, prior to cocultivation, slightly increased the number of β-glucuronidase expressing zones measured immediately after cocultivation, but it drastically decreased the number of transformed calli appearing on the explants 6 weeks after infection. Other factors examined were: Agrobacterium cell density during infection, bacterial growth phase, nature of the carbon source, explant age, and explant genotype.

  8. Enzymatic Synthesis of Biobased Polyesters and Polyamides

    Directory of Open Access Journals (Sweden)

    Yi Jiang


    Full Text Available Nowadays, “green” is a hot topic almost everywhere, from retailers to universities to industries; and achieving a green status has become a universal aim. However, polymers are commonly considered not to be “green”, being associated with massive energy consumption and severe pollution problems (for example, the “Plastic Soup” as a public stereotype. To achieve green polymers, three elements should be entailed: (1 green raw materials, catalysts and solvents; (2 eco-friendly synthesis processes; and (3 sustainable polymers with a low carbon footprint, for example, (biodegradable polymers or polymers which can be recycled or disposed with a gentle environmental impact. By utilizing biobased monomers in enzymatic polymerizations, many advantageous green aspects can be fulfilled. For example, biobased monomers and enzyme catalysts are renewable materials that are derived from biomass feedstocks; enzymatic polymerizations are clean and energy saving processes; and no toxic residuals contaminate the final products. Therefore, synthesis of renewable polymers via enzymatic polymerizations of biobased monomers provides an opportunity for achieving green polymers and a future sustainable polymer industry, which will eventually play an essential role for realizing and maintaining a biobased and sustainable society.

  9. Seismic air gun exposure during early-stage embryonic development does not negatively affect spiny lobster Jasus edwardsii larvae (Decapoda: Palinuridae). (United States)

    Day, Ryan D; McCauley, Robert D; Fitzgibbon, Quinn P; Semmens, Jayson M


    Marine seismic surveys are used to explore for sub-seafloor oil and gas deposits. These surveys are conducted using air guns, which release compressed air to create intense sound impulses, which are repeated around every 8-12 seconds and can travel large distances in the water column. Considering the ubiquitous worldwide distribution of seismic surveys, the potential impact of exposure on marine invertebrates is poorly understood. In this study, egg-bearing female spiny lobsters (Jasus edwardsii) were exposed to signals from three air gun configurations, all of which exceeded sound exposure levels (SEL) of 185 dB re 1 μPa(2) · s. Lobsters were maintained until their eggs hatched and the larvae were then counted for fecundity, assessed for abnormal morphology using measurements of larval length and width, tested for larval competency using an established activity test and measured for energy content. Overall there were no differences in the quantity or quality of hatched larvae, indicating that the condition and development of spiny lobster embryos were not adversely affected by air gun exposure. These results suggest that embryonic spiny lobster are resilient to air gun signals and highlight the caution necessary in extrapolating results from the laboratory to real world scenarios or across life history stages.

  10. Do parent-child acculturation gaps affect early adolescent Latino alcohol use? A study of the probability and extent of use. (United States)

    Cox, Ronald B; Roblyer, Martha Zapata; Merten, Michael J; Shreffler, Karina M; Schwerdtfeger, Kami L


    The literature has been mixed regarding how parent-child relationships are affected by the acculturation process and how this process relates to alcohol use among Latino youth. The mixed results may be due to, at least, two factors: First, staggered migration in which one or both parents arrive to the new country and then send for the children may lead to faster acculturation in parents than in children for some families. Second, acculturation may have different effects depending on which aspects of alcohol use are being examined. This study addresses the first factor by testing for a curvilinear trend in the acculturation-alcohol use relationship and the second by modeling past year alcohol use as a zero inflated negative binomial distribution. Additionally, this study examined the unique and mediation effects of parent-child acculturation discrepancies (gap), mother involvement in children's schooling, father involvement in children's schooling, and effective parenting on youth alcohol use during the last 12 months, measured as the probability of using and the extent of use. Direct paths from parent-child acculturation discrepancy to alcohol use, and mediated paths through mother involvement, father involvement, and effective parenting were also tested. Only father involvement fully mediated the path from parent-child acculturation discrepancies to the probability of alcohol use. None of the variables examined mediated the path from parent-child acculturation discrepancies to the extent of alcohol use. Effective parenting was unrelated to acculturation discrepancies; however, it maintained a significant direct effect on the probability of youth alcohol use and the extent of use after controlling for mother and father involvement. Implications for prevention strategies are discussed.

  11. Study of cassava starch enzymatic modification for maltodextrins obtention

    Directory of Open Access Journals (Sweden)

    Miguel Angel Díaz


    Full Text Available It was pretended to investigate the effect of the most relevant variables in cassava starch enzymatic hydrolysis process, on laboratory scale, to determine appropriate industrial conditions for the obtention of different kinds of maltodextrins. An a-Amylase enzyme, from genetically modified strain of Bacillus lichenijormis, was used to hydrolize the starch. Once the variables were chosen, an experimental fractioned factorial design was established with two levels. The Dextrose Equivalent (DE was taken as the response variable. The variables studied were: temperature (80-90°C, pH (5.5-6.5, dry-weight starch concentration (30-40% w/w, enzyme dosage (0.583-0.833 ul/g starch and calcium concentration (50-70 mgIL of CaClJ Some assays were settle in order to define the rheologic behavior and to determine the most relevant variables that affect the functional properties of the maltodextrin suspensions. Both, temperature and enzyme concentration were the variables that affect the most the initial convertion rate and the characteristics of the product convertion. For the interval of time considered, the highest DE obtained was 30 and the highest initial rate was 21.7 DE/hour. The functional properties of the maltodextrin solutions were affected by temperature and dry-weight starch concentration.

  12. Enzymatic activity measured by microcalorimetry in soil amended with organic residues

    Directory of Open Access Journals (Sweden)

    Karina Cenciani


    Full Text Available Enzymatic activity is an important property for soil quality evaluation. Two sequences of experiments were carried out in order to evaluate the enzymatic activity in a soil (Rhodic Eutrudox amended with cattle manure, earthworm casts, or sewage sludges from the municipalities of Barueri and Franca. The activity of commercial enzymes was measured by microcalorimetry in the same soil samples after sterilization. In the first experiment, the enzyme activities of cellulase, protease, and urease were determined in the soil samples during a three month period. In the second sequence of experiments, the thermal effect of the commercial enzymes cellulase, protease, and urease on sterilized soil samples under the same tretaments was monitored for a period of 46 days. The experimental design was randomized and arranged as factorial scheme in five treatments x seven samplings with five replications. The treatment effects were statistically evaluated by one-way analysis of variance. Tukey´s test was used to compare means at p < 0.05. The presence of different sources of organic residues increased the enzymatic activity in the sampling period. Cattle manure induced the highest enzymatic activity, followed by municipal sewage sludge, whereas earthworm casts induced the lowest activity, but differed from control treatment. The thermal effect on the enzyme activity of commercial cellulase, protease, and urease showed a variety of time peaks. These values probably oscillated due to soil physical-chemical factors affecting the enzyme activity on the residues.

  13. Pretreatment of corn stover using wet oxidation to enhance enzymatic digestibility. (United States)

    Varga, Eniko; Schmidt, Anette S; Réczey, Kati; Thomsen, Anne Belinda


    Corn stover is an abundant, promising raw material for fuel ethanol production. Although it has a high cellulose content, without pretreatment it resists enzymatic hydrolysis, like most lignocellulosic materials. Wet oxidation (water, oxygen, mild alkali or acid, elevated temperature and pressure) was investigated to enhance the enzymatic digestibility of corn stover. Six different combinations of reaction temperature, time, and pH were applied. The best conditions (60 g/L of corn stover, 195 degrees C, 15 min, 12 bar O2, 2 g/L of Na2CO3) increased the enzymatic conversion of corn stover four times, compared to untreated material. Under these conditions 60% of hemicellulose and 30% of lignin were solubilized, whereas 90% of cellulose remained in the solid fraction. After 24-h hydrolysis at 50 degrees C using 25 filter paper units (FPU)/g of drymatter (DM) biomass, the achieved conversion of cellulose to glucose was about 85%. Decreasing the hydrolysis temperature to 40 degrees C increased hydrolysis time from 24 to 72 h. Decreasing the enzyme loading to 5 FPU/g of DM biomass slightly decreased the enzymatic conversion from 83.4 to 71%. Thus, enzyme loading can be reduced without significantly affecting the efficiency of hydrolysis, an important economical aspect.

  14. Comparative performance of enzymatic and combined alkaline-enzymatic pretreatments on methane production from ensiled sorghum forage. (United States)

    Rollini, Manuela; Sambusiti, Cecilia; Musatti, Alida; Ficara, Elena; Retinò, Isabella; Malpei, Francesca


    This study investigated the effect of enzymatic and combined alkaline-enzymatic pretreatments on chemical composition and methane production from ensiled sorghum forage. Four commercial enzymatic preparations were tested and the two yielding the highest sugars release were added to evaluate any hydrolytic effect on both untreated and alkaline pretreated samples. In the combined alkaline-enzymatic pretreatment trials, the highest sugar release was found with Primafast and BGL preparations (added at a final concentration 0.12 and 0.20 mL/g TS, respectively), with a total monomeric content of 12 and 6.5 g/L. Fibre composition analysis confirmed that the combined alkaline-enzymatic pretreatment led to cellulose (up to 32 %) and hemicelluloses (up to 56 %) solubilisation, compared to the enzymatic pretreatment alone. BMP tests were performed on both untreated and pretreated samples, and time courses of methane production were fitted. Both enzymatic and combined alkaline-enzymatic pretreatment led to a methane production increase (304 and 362 mL CH4/g VS), compared to that of untreated sorghum (265 mL CH4/g VS), as  +15 and  +37 %, respectively. Moreover, higher specific methane production rates, compared to that of untreated sorghum (20.31 mL CH4/g VS/d), were obtained by applying the enzymatic and combined alkaline-enzymatic pretreatment (33.94 and 31.65 mL CH4/g VS/d), respectively.

  15. Enzymatic Hydrolysis of Alkaline Pretreated Coconut Coir

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    Akbarningrum Fatmawati


    Full Text Available The purpose of this research is to study the effect of concentration and temperature on the cellulose and lignin content, and the reducing sugars produced in the enzymatic hydrolysis of coconut coir. In this research, the coconut coir is pretreated using 3%, 7%, and 11% NaOH solution at 60oC, 80oC, and 100oC. The pretreated coir were assayed by measuring the amount of cellulose and lignin and then hydrolysed using Celluclast and Novozyme 188 under various temperature (30oC, 40oC, 50oC and pH (3, 4, 5. The hydrolysis results were assayed for the reducing sugar content. The results showed that the alkaline delignification was effective to reduce lignin and to increase the cellulose content of the coir. The best delignification condition was observed at 11% NaOH solution and 100oC which removed 14,53% of lignin and increased the cellulose content up to 50,23%. The best condition of the enzymatic hydrolysis was obtained at 50oC and pH 4 which produced 7,57 gr/L reducing sugar. © 2013 BCREC UNDIP. All rights reservedReceived: 2nd October 2012; Revised: 31st January 2013; Accepted: 6th February 2013[How to Cite: Fatmawati, A., Agustriyanto, R., Liasari, Y. (2013. Enzymatic Hydrolysis of Alkaline Pre-treated Coconut Coir. Bulletin of Chemical Reaction Engineering & Catalysis, 8 (1: 34-39 (doi:10.9767/bcrec.8.1.4048.34-39[Permalink/DOI:] | View in  |

  16. Biodegradation of Leather Waste by Enzymatic Treatment

    Institute of Scientific and Technical Information of China (English)


    The treatment of shavings, trimmings and splits of leather waste from tanneries has a potential to generate value-added products. In this study enzymatic treatment of leather waste was performed. This method utilizes alkaline protease produced by Bacillus subtilis in our laboratory by submerged fermentation. Optimum conditions of pH, time duration,temperature and concentration of enzyme were determined for maximum degradation of leather waste. The amount of degradation was measured by the release of amino acid hydroxyproline. Amino acid composition in the hydrolysate obtained by the enzyme hydrolysis was determined. This relative simple biotreatment of leather waste may provide a practical and economical solution.

  17. The Enzymatic Antioxidant System of Human Spermatozoa

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    Cristian O’Flaherty


    Full Text Available The ejaculated spermatozoon, as an aerobic cell, must fight against toxic levels of reactive oxygen species (ROS generated by its own metabolism but also by other sources such as abnormal spermatozoa, chemicals and toxicants, or the presence of leukocytes in semen. Mammalian spermatozoa are extremely sensitive to oxidative stress, a condition occurring when there is a net increase in ROS levels within the cell. Opportunely, this specialized cell has a battery of antioxidant enzymes (superoxide dismutase, peroxiredoxins, thioredoxins, thioredoxins reductases, and glutathione s-transferases working in concert to assure normal sperm function. Any impairment of the antioxidant enzymatic activities will promote severe oxidative damage which is observed as plasma membrane lipid peroxidation, oxidation of structural proteins and enzymes, and oxidation of DNA bases that lead to abnormal sperm function. Altogether, these damages occurring in spermatozoa are associated with male infertility. The present review contains a description of the enzymatic antioxidant system of the human spermatozoon and a reevaluation of the role of its different components and highlights the necessity of sufficient supply of reducing agents (NADPH and reduced glutathione to guarantee normal sperm function.

  18. Enzymatic and bacterial conversions during sourdough fermentation. (United States)

    Gänzle, Michael G


    Enzymatic and microbial conversion of flour components during bread making determines bread quality. Metabolism of sourdough microbiota and the activity of cereal enzymes are interdependent. Acidification, oxygen consumption, and thiols accumulation by microbial metabolism modulate the activity of cereal enzymes. In turn, cereal enzymes provide substrates for bacterial growth. This review highlights the role of cereal enzymes and the metabolism of lactic acid bacteria in conversion of carbohydrates, proteins, phenolic compounds and lipids. Heterofermentative lactic acid bacteria prevailing in wheat and rye sourdoughs preferentially metabolise sucrose and maltose; the latter is released by cereal enzymes during fermentation. Sucrose supports formation of acetate by heterofermentative lactobacilli, and the formation of exopolysaccharides. The release of maltose and glucose by cereal enzymes during fermentation determines the exopolysaccharide yield in sourdough fermentations. Proteolysis is dependent on cereal proteases. Peptidase activities of sourdough lactic acid bacteria determine the accumulation of (bioactive) peptides, amino acids, and amino acid metabolites in dough and bread. Enzymatic conversion and microbial metabolism of phenolic compounds is relevant in sorghum and millet containing high levels of phenolic compounds. The presence of phenolic compounds with antimicrobial activity in sorghum selects for fermentation microbiota that are resistant to the phenolic compounds.

  19. Palm Date Fibers: Analysis and Enzymatic Hydrolysis

    Directory of Open Access Journals (Sweden)

    Mohammad J. Taherzadeh


    Full Text Available Waste palm dates were subjected to analysis for composition and enzymatic hydrolysis of their flesh fibers. The fruit contained 32% glucose and 30% fructose, while the water-insoluble fibers of its flesh consisted of 49.9% lignin and 20.9% polysaccharides. Water-insoluble fibers were settled to 55% of its initial volume in 12 h. The presence of skin and flesh colloidal fibers results in high viscosity and clogging problems during industrial processes. The settling velocity of the fibers was improved by enzymatic hydrolysis. Hydrolysis resulted in 84.3% conversion of the cellulosic part of the fibers as well as reducing the settling time to 10 minutes and the final settled volume to 4% of the initial volume. It implies easier separation of the fibers and facilitates fermentation processes in the corresponding industries. Two kinds of high- and low-lignin fibers were identified from the water-insoluble fibers. The high-lignin fibers (75% lignin settled easily, while the low-lignin fibers (41.4% lignin formed a slurry suspension which settled very slowly. The hydrophilicity of these low-lignin fibers is the major challenge of the industrial processes.

  20. Microfluidic enzymatic biosensing systems: A review. (United States)

    Mross, Stefan; Pierrat, Sebastien; Zimmermann, Tom; Kraft, Michael


    Microfluidic biosensing systems with enzyme-based detection have been extensively studied in the last years owing to features such as high specificity, a broad range of analytes and a high degree of automation. This review gives an overview of the most important factors associated with these systems. In the first part, frequently used immobilization protocols such as physisorption and covalent bonding and detection techniques such as amperometry and fluorescence measurements are discussed with respect to effort, lifetime and measurement range. The Michaelis-Menten model describing the kinetics of enzymatic reactions, the role of redox mediators and the limitations of the linear measurement range of enzymatic sensors are introduced. Several possibilities of extending the linear measurement range in microfluidic systems such as diffusion-limiting membranes and the flow injection setup are presented. Regarding the integration of enzymes into microfluidic systems during the fabrication process, the constraints imposed by the biomolecules due to the limited usage of high temperatures and solvents are addressed. In the second part, the most common forms of enzyme integration into microfluidic systems, i.e. in channels and on electrodes, on microparticles, on paper and thread and as injected enzyme solutions, are reviewed, focusing on fabrication, applications and performance.

  1. Fluorometric enzymatic assay of L-arginine (United States)

    Stasyuk, Nataliya; Gayda, Galina; Yepremyan, Hasmik; Stepien, Agnieszka; Gonchar, Mykhailo


    The enzymes of L-arginine (further - Arg) metabolism are promising tools for elaboration of selective methods for quantitative Arg analysis. In our study we propose an enzymatic method for Arg assay based on fluorometric monitoring of ammonia, a final product of Arg splitting by human liver arginase I (further - arginase), isolated from the recombinant yeast strain, and commercial urease. The selective analysis of ammonia (at 415 nm under excitation at 360 nm) is based on reaction with o-phthalaldehyde (OPA) in the presence of sulfite in alkali medium: these conditions permit to avoid the reaction of OPA with any amino acid. A linearity range of the fluorometric arginase-urease-OPA method is from 100 nM to 6 μМ with a limit of detection of 34 nM Arg. The method was used for the quantitative determination of Arg in the pooled sample of blood serum. The obtained results proved to be in a good correlation with the reference enzymatic method and literature data. The proposed arginase-urease-OPA method being sensitive, economical, selective and suitable for both routine and micro-volume formats, can be used in clinical diagnostics for the simultaneous determination of Arg as well as urea and ammonia in serum samples.

  2. Enzymatic hydrolysis of lactose of whey permeate

    Directory of Open Access Journals (Sweden)

    Karina Nascimento de Almeida


    Full Text Available The whey permeate is the residual of the concentration process of the whey proteins by ultrafiltration method. It contains important nutrients such as lactose, minerals and some proteins and lipids. It is without an ending industrial waste that causes serious damage to the environment. For its full use the lactose must be hydrolyzed to enable its consumption by intolerant people. The enzymatic hydrolysis by lactase (β-galactosidase of Kluyveromyces lactis yeast is a safe method that does not compromise the integrity of other nutrients, enabling further use of the permeate as a raw material. This study aimed to perform tests of enzymatic hydrolysis of lactose in whey permeate formulations in a concentration of 0.2%, 0.7% and 1% at 30, 60 and 90 minutes with pH 6.3 medium and 37 °C. The reactions were monitored by high performance liquid chromatography which showed that the enzyme concentration of 0.7% at time 30 minutes formulations became safe for consumption by lactose intolerant people, according to minimum levels established by law.

  3. Conditional iron and pH-dependent activity of a non-enzymatic glycolysis and pentose phosphate pathway. (United States)

    Keller, Markus A; Zylstra, Andre; Castro, Cecilia; Turchyn, Alexandra V; Griffin, Julian L; Ralser, Markus


    Little is known about the evolutionary origins of metabolism. However, key biochemical reactions of glycolysis and the pentose phosphate pathway (PPP), ancient metabolic pathways central to the metabolic network, have non-enzymatic pendants that occur in a prebiotically plausible reaction milieu reconstituted to contain Archean sediment metal components. These non-enzymatic reactions could have given rise to the origin of glycolysis and the PPP during early evolution. Using nuclear magnetic resonance spectroscopy and high-content metabolomics that allowed us to measure several thousand reaction mixtures, we experimentally address the chemical logic of a metabolism-like network constituted from these non-enzymatic reactions. Fe(II), the dominant transition metal component of Archean oceanic sediments, has binding affinity toward metabolic sugar phosphates and drives metabolism-like reactivity acting as both catalyst and cosubstrate. Iron and pH dependencies determine a metabolism-like network topology and comediate reaction rates over several orders of magnitude so that the network adopts conditional activity. Alkaline pH triggered the activity of the non-enzymatic PPP pendant, whereas gentle acidic or neutral conditions favored non-enzymatic glycolytic reactions. Fe(II)-sensitive glycolytic and PPP-like reactions thus form a chemical network mimicking structural features of extant carbon metabolism, including topology, pH dependency, and conditional reactivity. Chemical networks that obtain structure and catalysis on the basis of transition metals found in Archean sediments are hence plausible direct precursors of cellular metabolic networks.

  4. Enzymatic activity of cell-free extracts from Burkholderia oxyphila OX-01 bio-converts (+)-catechin and (-)-epicatechin to (+)-taxifolin. (United States)

    Otsuka, Yuichiro; Matsuda, Motoki; Sonoki, Tomonori; Sato-Izawa, Kanna; Goodell, Barry; Jelison, Jody; Navarro, Ronald R; Murata, Hitoshi; Nakamura, Masaya


    This study characterized the enzymatic ability of a cell-free extract from an acidophilic (+)-catechin degrader Burkholderia oxyphila (OX-01). The crude OX-01 extracts were able to transform (+)-catechin and (-)-epicatechin into (+)-taxifolin via a leucocyanidin intermediate in a two-step oxidation. Enzymatic oxidation at the C-4 position was carried out anaerobically using H2O as an oxygen donor. The C-4 oxidation occurred only in the presence of the 2R-catechin stereoisomer, with the C-3 stereoisomer not affecting the reaction. These results suggest that the OX-01 may have evolved to target both (+)-catechin and (-)-epicatechin, which are major structural units in plants.

  5. Early harvest affects ratooning ability in Louisiana (United States)

    Similar to sugarcane industries around the world, economies of scale often lead to decreases in total number of manufacturing centers such as sugar mills. One of the consequences of mill closures has been an increase in the duration of the crushing season in Louisiana, which has historically bee...

  6. Inhibition of cellulose enzymatic hydrolysis by laccase-derived compounds from phenols. (United States)

    Oliva-Taravilla, Alfredo; Tomás-Pejó, Elia; Demuez, Marie; González-Fernández, Cristina; Ballesteros, Mercedes


    The presence of inhibitors compounds after pretreatment of lignocellulosic materials affects the saccharification and fermentation steps in bioethanol production processes. Even though, external addition of laccases selectively removes the phenolic compounds from lignocellulosic prehydrolysates, when it is coupled to saccharification step, lower hydrolysis yields are attained. Vanillin, syringaldehyde and ferulic acid are phenolic compounds commonly found in wheat-straw prehydrolysate after steam-explosion pretreatment. These three phenolic compounds were used in this study to elucidate the inhibitory mechanisms of laccase-derived compounds after laccase treatment. Reaction products derived from laccase oxidation of vanillin and syringaldehyde showed to be the strongest inhibitors. The presence of these products causes a decrement on enzymatic hydrolysis yield of a model cellulosic substrate (Sigmacell) of 46.6 and 32.6%, respectively at 24 h. Moreover, a decrease in more than 50% of cellulase and β-glucosidase activities was observed in presence of laccase and vanillin. This effect was attributed to coupling reactions between phenoxyl radicals and enzymes. On the other hand, when the hydrolysis of Sigmacell was performed in presence of prehydrolysate from steam-exploded wheat straw a significant inhibition on enzymatic hydrolysis was observed independently of laccase treatment. This result pointed out that the other components of wheat-straw prehydrolysate are affecting the enzymatic hydrolysis to a higher extent than the possible laccase-derived products.

  7. Impact of Enzymatic Degradation of the Endothelial Glycocalyx on Vascular Permeability in an Awake Hamster Model

    Directory of Open Access Journals (Sweden)

    S. A. Landsverk


    Full Text Available Background. The inside of the endothelium is covered by a glycocalyx layer, and enzymatic degradation of this layer induces vascular leakage ex vivo. We hypothesized that enzymatic degrading of the glycocalyx in an in vivo, whole body model, would induce plasma leakage and affect the microcirculation. Methods. Golden Syrian hamsters were divided into an enzyme (hyaluronidase and a control group. Mean arterial pressure (MAP, heart rate (HR, hematocrit (Hct, base excess (BE, and plasma volume were obtained before, 45 and 120 min after enzyme/saline treatment. Plasma volume was evaluated by the distribution volume of indocyanine green and the microcirculation by functional capillary density (FCD. The enzymatic effect was determined by measuring plasma levels of hyaluronan (HA. Results. There were no differences in MAP, HR, Hct, and BE between the two groups. Enzyme treatment did not induce changes in plasma volume but reduced FCD. There was a 50–100-fold increase in plasma HA, but no relationship was found between HA levels and plasma volume or FCD. Conclusion. Vascular leakage was not confirmed in an in vivo, whole body model after degradation of the endothelial glycocalyx. The microcirculation was affected, but no relationship between plasma levels of HA and FCD was seen.

  8. Pretreatment and enzymatic hydrolysis of lignocellulosic biomass (United States)

    Corredor, Deisy Y.

    The performance of soybean hulls and forage sorghum as feedstocks for ethanol production was studied. The main goal of this research was to increase fermentable sugars' yield through high-efficiency pretreatment technology. Soybean hulls are a potential feedstock for production of bio-ethanol due to their high carbohydrate content (≈50%) of nearly 37% cellulose. Soybean hulls could be the ideal feedstock for fuel ethanol production, because they are abundant and require no special harvesting and additional transportation costs as they are already in the plant. Dilute acid and modified steam-explosion were used as pretreatment technologies to increase fermentable sugars yields. Effects of reaction time, temperature, acid concentration and type of acid on hydrolysis of hemicellulose in soybean hulls and total sugar yields were studied. Optimum pretreatment parameters and enzymatic hydrolysis conditions for converting soybean hulls into fermentable sugars were identified. The combination of acid (H2SO4, 2% w/v) and steam (140°C, 30 min) efficiently solubilized the hemicellulose, giving a pentose yield of 96%. Sorghum is a tropical grass grown primarily in semiarid and dry parts of the world, especially in areas too dry for corn. The production of sorghum results in about 30 million tons of byproducts mainly composed of cellulose, hemicellulose, and lignin. Forage sorghum such as brown midrib (BMR) sorghum for ethanol production has generated much interest since this trait is characterized genetically by lower lignin concentrations in the plant compared with conventional types. Three varieties of forage sorghum and one variety of regular sorghum were characterized and evaluated as feedstock for fermentable sugar production. Fourier transform infrared spectroscopy (FTIR), scanning electron microscope (SEM) and X-Ray diffraction were used to determine changes in structure and chemical composition of forage sorghum before and after pretreatment and enzymatic hydrolysis

  9. Enzymatic synthesis and NMR studies of acylated sucrose acetates

    NARCIS (Netherlands)

    Steverink-De Zoete, M.C.; Kneepkens, M.F.M.; Waard, de P.; Woudenberg-van Oosterom, M.; Gotlieb, K.F.; Slaghek, T.


    The lipase-catalyzed esterification of partially acetylated sucrose has been studied. It was shown that the chemical acetylation increased the reaction rate of the subsequent enzymatic acylation. Thus it was possible to perform the enzymatic acylation in the absence of solvents while underivatized s

  10. Physical properties and structure of enzymatically synthesized amylopectin analogs

    NARCIS (Netherlands)

    Ciric, Jelena; Woortman, Albert J. J.; Gordiichuk, Pavlo; Stuart, Marc C. A.; Loos, Katja


    The mechanism of the enzymatic polymerization of amylopectin analogs with phosphorylase b and glycogen branching enzyme is very intriguing. Recently, size exclusion chromatography with multi-detection of enzymatically synthesized amylopectin analogs in combination with MALDI-ToF MS analysis of enzym

  11. Inhibition of enzymatic browning in foods and beverages. (United States)

    McEvily, A J; Iyengar, R; Otwell, W S


    Enzymatic browning is a major factor contributing to quality loss in foods and beverages. Sulfiting agents are used commonly to control browning; however, several negative attributes associated with sulfites have created the need for functional alternatives. Recent advances in the development of nonsulfite inhibitors of enzymatic browning are reviewed. The review focuses on compositions that are of practical relevance to food use.

  12. Modelling the Effects of Ageing Time of Starch on the Enzymatic Activity of Three Amylolytic Enzymes

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    Nelson P. Guerra


    Full Text Available The effect of increasing ageing time (t of starch on the activity of three amylolytic enzymes (Termamyl, San Super, and BAN was investigated. Although all the enzymatic reactions follow michaelian kinetics, vmax decreased significantly (P<0.05 and KM increased (although not always significantly with the increase in t. The conformational changes produced in the starch chains as a consequence of the ageing seemed to affect negatively the diffusivity of the starch to the active site of the enzymes and the release of the reaction products to the medium. A similar effect was observed when the enzymatic reactions were carried out with unaged starches supplemented with different concentrations of gelatine [G]. The inhibition in the amylolytic activities was best mathematically described by using three modified forms of the Michaelis-Menten model, which included a term to consider, respectively, the linear, exponential, and hyperbolic inhibitory effects of t and [G].

  13. Lipophilic prodrugs of apomorphine I: Preparation, characterisation, and in vitro enzymatic hydrolysis in biorelevant media

    DEFF Research Database (Denmark)

    Borkar, Nrupa Nitin; Li, Boyang; Holm, René;


    in biorelevant media before and after incorporating them into self-emulsifying drug delivery systems (SEDDS) for oral delivery. Two apomorphine diester prodrugs were synthesised: dilauroyl apomorphine (DLA) and dipalmitoyl apomorphine (DPA). The in vitro enzymatic hydrolysis of diesters was performed using...... biorelevant media with pancreatin to catalyse the diester degradation. The synthesised and purified diesters were found to be free from reactants as impurities confirmed by LC/MS and NMR. DLA and DPA were degraded into corresponding monoesters and free apomorphine within 5 min after adding pancreatin, leaving...... about 4% and 28% of the intact diester, respectively. The incorporation of the diesters into SEDDS reduced the enzymatic degradation of diesters. In addition, the chain length of diester and the type of oil used in formulations affected diester hydrolysis. The lipophilic apomorphine diesters were...

  14. Non-Enzymatic Graphene-Based Biosensors for Continous Glucose Monitoring

    CERN Document Server

    Serry, Mohamed


    A novel mediator-free, non-enzymatic electrochemical sensor, based on a graphene-Schottky junction, was fabricated for glucose detection. The sensor offers a promising alternative to the conventional enzyme-catalyzed electrochemical continuous glucose monitoring systems (CGM), as it overcomes many of the drawbacks attributed to the enzymatic nature; namely, irreversibility, drift, and interference with body fluids, which affect their accuracy, reliability and longevity. Enhanced performance of the sensors is demonstrated through the band interaction at the graphene-Schottky junction, which yields stronger forward/reverse currents in response to 50 {\\mu}L glucose drop. Under optimized conditions, the linear response of the sensor to glucose concentration was valid in the range from 0 to 15 mmol/L with a detection limit of 0.5 mmol/L. The results indicated that the proposed sensor provided a highly sensitive, more facile method with good reproducibility for continuous glucose detection.

  15. Dynamic Simulation, Sensitivity and Uncertainty Analysis of a Demonstration Scale Lignocellulosic Enzymatic Hydrolysis Process

    DEFF Research Database (Denmark)

    Prunescu, Remus Mihail; Sin, Gürkan


    , accounting a large number of parameters. The sensitivity analysis of model predictions with respect to model parameters is quantified by the delta mean square measure. By ranking the delta mean square, a reduced subset of parameters is found helping to identify the bottleneck of the model. The uncertainty...... to be affected by non-zero mean noise because they are determined by a Near Infrared (NIR) instrument. LHS is performed on 2 parameters: the probability of the mean value and the probability of the standard deviation for each measurement. The Monte Carlo outputs are then analyzed by linear regression......This study presents the uncertainty and sensitivity analysis of a lignocellulosic enzymatic hydrolysis model considering both model and feed parameters as sources of uncertainty. The dynamic model is parametrized for accommodating various types of biomass, and different enzymatic complexes...

  16. Structure and enzymatic accessibility of leaf and stem from wheat straw before and after hydrothermal pretreatment

    DEFF Research Database (Denmark)

    Zhang, Heng; Thygesen, Lisbeth Garbrecht; Mortensen, Kell;


    Background: Biomass recalcitrance is affected by a number of chemical, physical and biological factors. In this study we looked into the differences in recalcitrance between two major anatomical fractions of wheat straw biomass, leaf and stem. A set of twenty-one wheat cultivars was fractionated...... and illustrated the substantial variation in leaf-to-stem ratio between cultivars. The two fractions were compared in terms of chemical composition, enzymatic convertibility, cellulose crystallinity and glucan accessibility. The use of water as a probe for assessing glucan accessibility was explored using low....... Hydrothermal pretreatment increased the accessibility towards water in stems but not in leaves. The results in this study indicate a correlation between the accessibility of glucan to water and to enzymes. Conclusions: Enzymatic degradability of wheat straw anatomical fractions can be indicated...

  17. Phosphoric acid pretreatment of Achyranthes aspera and Sida acuta weed biomass to improve enzymatic hydrolysis. (United States)

    Siripong, Premjet; Duangporn, Premjet; Takata, Eri; Tsutsumi, Yuji


    Achyranthes aspera and Sida acuta, two types of weed biomass are abundant and waste in Thailand. We focus on them as novel feedstock for bio-ethanol production because they contain high-cellulose content (45.9% and 46.9%, respectively) and unutilized material. Phosphoric acid (70%, 75%, and 80%) was employed for the pretreatment to improve by enzymatic hydrolysis. The pretreatment process removed most of the xylan and a part of the lignin from the weeds, while most of the glucan remained. The cellulose conversion to glucose was greater for pretreated A. aspera (86.2 ± 0.3%) than that of the pretreated S. acuta (82.2 ± 1.1%). Thus, the removal of hemicellulose significantly affected the efficiency of the enzymatic hydrolysis. The scanning electron microscopy images showed the exposed fibrous cellulose on the cell wall surface, and this substantial change of the surface structure contributed to improving the enzyme accessibility.

  18. Structural basis of enzymatic benzene ring reduction. (United States)

    Weinert, Tobias; Huwiler, Simona G; Kung, Johannes W; Weidenweber, Sina; Hellwig, Petra; Stärk, Hans-Joachim; Biskup, Till; Weber, Stefan; Cotelesage, Julien J H; George, Graham N; Ermler, Ulrich; Boll, Matthias


    In chemical synthesis, the widely used Birch reduction of aromatic compounds to cyclic dienes requires alkali metals in ammonia as extremely low-potential electron donors. An analogous reaction is catalyzed by benzoyl-coenzyme A reductases (BCRs) that have a key role in the globally important bacterial degradation of aromatic compounds at anoxic sites. Because of the lack of structural information, the catalytic mechanism of enzymatic benzene ring reduction remained obscure. Here, we present the structural characterization of a dearomatizing BCR containing an unprecedented tungsten cofactor that transfers electrons to the benzene ring in an aprotic cavity. Substrate binding induces proton transfer from the bulk solvent to the active site by expelling a Zn(2+) that is crucial for active site encapsulation. Our results shed light on the structural basis of an electron transfer process at the negative redox potential limit in biology. They open the door for biological or biomimetic alternatives to a basic chemical synthetic tool.

  19. Isothermal calorimetry on enzymatic biodiesel production

    DEFF Research Database (Denmark)

    Fjerbæk, Lene


    information about effects taking place when using lipases immobilized on an inert carrier for transesterification of a triglyceride and an alcohol as for biodiesel production. The biodiesel is produced by rapeseed oil and methanol as well as ethanol and a commercial biocatalyst Novozym 435 from Novozymes...... containing a Candida Antarctica B lipase immobilized on an acrylic resin. The reaction investigated is characterized by immiscible liquids (oil, methanol, glycerol and biodiesel) and enzymes imm. on an inert carrier during reaction, which allows several effects to take place that during normal reaction...... conditions can not be elucidated. These effects have been observed with isothermal calorimetry bringing forth new information about the reaction of enzymes catalyzing transesterification. Enzymatic biodiesel production has until now not been investigated with isothermal microcalorimetry, but the results...

  20. Extracellular enzymatic activities of Bipolaris sorokiniana isolates. (United States)

    Geimba, Mercedes P; Brandelli, Adriano


    Several enzymatic activities were investigated in six isolates of the fungus Bipolaris sorokiniana, originating from different areas of Brazil. Among the glycosidases studied, beta-glucosidase, beta-N-acetylglucosaminidase, beta-xylosidase, cellobiohydrolase, and chitobiohydrolase were the major activities. In some isolates, beta-glucuronidase, beta-galactosidase, and alpha-mannosidase activities were also present. Polysaccharide-hydrolyzing enzymes, such as pectin lyase and carboxymethyl cellulase were detected in significant amounts, and their activities were variable among the different isolates. Other enzymes, namely phosphatases, proteinases and phenol oxidase, were also examined, showing variable amounts depending on the isolate. The pH dependence of all enzymes tested was investigated. Endoproteinase, carboxymethyl cellulase, and phenoloxidase had maximum activity in the pH range of 6-8, whilst all other enzymes showed maximum activity at pH 4-6.

  1. Cascade enzymatic reactions for efficient carbon sequestration. (United States)

    Xia, Shunxiang; Zhao, Xueyan; Frigo-Vaz, Benjamin; Zheng, Wenyun; Kim, Jungbae; Wang, Ping


    Thermochemical processes developed for carbon capture and storage (CCS) offer high carbon capture capacities, but are generally hampered by low energy efficiency. Reversible cascade enzyme reactions are examined in this work for energy-efficient carbon sequestration. By integrating the reactions of two key enzymes of RTCA cycle, isocitrate dehydrogenase and aconitase, we demonstrate that intensified carbon capture can be realized through such cascade enzymatic reactions. Experiments show that enhanced thermodynamic driving force for carbon conversion can be attained via pH control under ambient conditions, and that the cascade reactions have the potential to capture 0.5 mol carbon at pH 6 for each mole of substrate applied. Overall it manifests that the carbon capture capacity of biocatalytic reactions, in addition to be energy efficient, can also be ultimately intensified to approach those realized with chemical absorbents such as MEA.

  2. Enzymatic degradation of multiwalled carbon nanotubes. (United States)

    Zhao, Yong; Allen, Brett L; Star, Alexander


    Because of their unique properties, carbon nanotubes and, in particular, multiwalled carbon nanotubes (MWNTs) have been used for the development of advanced composite and catalyst materials. Despite their growing commercial applications and increased production, the potential environmental and toxicological impacts of MWNTs are not fully understood; however, many reports suggest that they may be toxic. Therefore, a need exists to develop protocols for effective and safe degradation of MWNTs. In this article, we investigated the effect of chemical functionalization of MWNTs on their enzymatic degradation with horseradish peroxidase (HRP) and hydrogen peroxide (H(2)O(2)). We investigated HRP/H(2)O(2) degradation of purified, oxidized, and nitrogen-doped MWNTs and proposed a layer-by-layer degradation mechanism of nanotubes facilitated by side wall defects. These results provide a better understanding of the interaction between HRP and carbon nanotubes and suggest an eco-friendly way of mitigating the environmental impact of nanotubes.

  3. Heavy atom isotope effects on enzymatic reactions (United States)

    Paneth, Piotr


    The theory of isotope effects, which has proved to be extremely useful in providing geometrical details of transition states in a variety of chemical reactions, has recently found an application in studies of enzyme-catalyzed reactions. These reactions are multistep in nature with few steps being partially rate-limiting, thus interpretation of these isotope effects is more complex. The theoretical framework of heavy-atom isotope effects on enzymatic reactions is critically analyzed on the basis of recent results of: carbon kinetic isotope effects on carbonic anhydrase and catalytic antibodies; multiple carbon, deuterium isotope effects on reactions catalyzed by formate decarboxylase; oxygen isotope effects on binding processes in reactions catalyzed by pyruvate kinase; and equilibrium oxygen isotope effect on binding an inhibitor to lactate dehydrogenase. The advantages and disadvantages of reaction complexity in learning details of formal and molecular mechanisms are discussed in the examples of reactions catalyzed by phosphoenolpyruvate carboxylase, orotidine decarboxylase and glutamine synthetase.

  4. Isothermal Titration Calorimetry to Characterize Enzymatic Reactions. (United States)

    Mazzei, Luca; Ciurli, Stefano; Zambelli, Barbara


    Isothermal titration calorimetry (ITC) is a technique that measures the heat released or absorbed during a chemical reaction as an intrinsic probe to characterize any chemical process that involves heat changes spontaneously occurring during the reaction. The general features of this method to determine the kinetic and thermodynamic parameters of enzymatic reactions (kcat, KM, ΔH) are described and discussed here together with some detailed applications to specific cases. ITC does not require any modification or labeling of the system under analysis, can be performed in solution, and needs only small amounts of enzyme. These properties make ITC an invaluable, powerful, and unique tool to extend the knowledge of enzyme kinetics to drug discovery.

  5. Structure of the enzymatically synthesized fructan inulin

    Energy Technology Data Exchange (ETDEWEB)

    Heyer, A.G.; Schroeer, B. [Max-Planck-Institut fuer Molekulare Pflanzenphysiologie, Karl-Liebknecht-Str. 25, 14476 Golm (Germany); Radosta, S. [Fraunhofer-Institut fuer Angewandte Polymerforschung, Postfach 126, 14504 Teltow (Germany); Wolff, D.; Czapla, S.; Springer, J. [Technische Universitaet Berlin, FG Makromolekulare Chemie, Str. des 17. Juni 135, 10623 Berlin (Germany)


    Construction, purification and characterization of a fusion protein of maltose-binding protein of Escherichia coli and the fructosyltransferase of Streptococcus mutans is described. With the purified protein, in vitro synthesis of inulin was performed. The obtained polysaccharide was characterized by high-performance size-exclusion chromatography (HPSEC) and static light scattering (SLS) in dilute aqueous and dimethyl sulfoxide solution. For all samples very high molecular weights between 60x10{sup 6} and 90x10{sup 6} g/mol and a remarkable small polydispersity index of 1.1 have been determined. Small root-mean-square radii of gyration point to a compact conformation in dilute solution. No difference between native and enzymatically synthesized inulin was observed by X-ray powder diffraction and thermoanalysis of solid samples. (Copyright (c) 1998 Elsevier Science B.V., Amsterdam. All rights reserved.)

  6. Thermal stability of bioactive enzymatic papers. (United States)

    Khan, Mohidus Samad; Li, Xu; Shen, Wei; Garnier, Gil


    The thermal stability of two enzymes adsorbed on paper, alkaline phosphatase (ALP) and horseradish peroxidase (HRP), was measured using a colorimetric technique quantifying the intensity of the product complex. The enzymes adsorbed on paper retained their functionality and selectivity. Adsorption on paper increased the enzyme thermal stability by 2-3 orders of magnitude compared to the same enzyme in solution. ALP and HRP enzymatic papers had half-lives of 533 h and 239 h at 23 degrees C, respectively. The thermal degradation of adsorbed enzyme was found to follow two sequential first-order reactions, indication of a reaction system. A complex pattern of enzyme was printed on paper using a thermal inkjet printer. Paper and inkjet printing are ideal material and process to manufacture low-cost-high volume bioactive surfaces.

  7. Recent development of miniatured enzymatic biofuel cell (United States)

    Song, Yin; Penmatsa, Varun; Wang, Chunlei


    Enzymatic biofuel cells (EBFCs) that oxidize biological fuels using enzyme-modified electrodes are considered a promising candidate for implantable power sources. However, there are still challenges to overcome before biofuel cells become competitive in any practical applications. Currently, the short lifespan of the catalytic enzymes and poor power density are the most critical issues in developing EBFCs. In this paper, we will review the recent development of biofuel cells and highlight the progress in Carbon-microelectromechanical system (C-MEMS) based micro biofuel cells by both computational modeling and experimental work. Also, our effort on utilizing a covalent immobilization technique for the attachment of enzymes onto the substrate which is expected to increase the enzyme loading efficiency and the power density of devices is discussed in this paper.

  8. Enzymatic assay for methotrexate in erythrocytes

    DEFF Research Database (Denmark)

    Schrøder, H; Heinsvig, E M


    Methotrexate (MTX) accumulates in erythrocytes in MTX-treated patients. We present a modified enzymatic assay measuring MTX concentrations between 10 and 60 nmol/l in erythrocytes, adapted for a centrifugal analyser (Cobas Bio). About 40 patient's samples could be analysed within 1 h. The detection...... limit was 3 nmol/l. Within run and between-run precision was 7.4% and 13.5% for control 10 nmol/l and 1.2% and 3.2% for control 50 nmol/l. Recovery was 85-115% of MTX added to haemolysed erythrocytes. We found the method useful for pharmacokinetic studies of MTX in erythrocytes in MTX-treated patients...

  9. Synthesis of monoacylglycerols by enzymatic methods

    Directory of Open Access Journals (Sweden)

    Bradić Milena R.


    Full Text Available Monoacylglycerols are non-ionic surfactants widely used in the food industry. They are also important in cosmetic and pharmaceutical industries as drug carriers and for the consistency improvements in creams and lotions. Current process for their production is based on the glycerolysis of natural fats and oils in the presence of inorganic catalysts at temperatures higher than 220 oC. The major drawbacks of this process include high-energy consumption, low yield, and poor product quality. The use of lipases for the monoacylglycerols production offers environmental advantages and a reduction in energy consumption. Besides, the same surfactants prepared by the enzymatic synthesis may be labeled as “natural”. Recent progress in the application of highly-stable lipases in the organic solvents offers the possibility of employing various methods to the enzyme-catalyzed synthesis of monoacylglycerols, such as selective hydrolysis of fats and oils using 1,3-regiospecific lipases, the esterification of glycerol with fatty acids and the glycerolysis of fats or oils. In this review, different reaction systems such as aqueous-organic two-phase systems, microemulsions and reverse micelles systems, anhydrous organic solvents, solvent-free systems with free or immobilized lipases, as well as the use of two-phase membrane reactor systems are presented. We discuss some of the key factors, such as the control of water content, removing of the products from reaction system, and the effects of solvent on the lipase activity and selectivity, that must be addressed in order to obtain an efficient reaction system with high yields of monoacylglycerols. Engineering of the enzymatic monoacylglycerols synthesis processes requires also optimization of other factors as: molar ratio of substrates, temperature, type of lipase immobilization and supports (if any, reactor design and operating regime.

  10. Characterizing Enzymatic Deposition for Microelectrode Neurotransmitter Detection

    Energy Technology Data Exchange (ETDEWEB)

    Hosein, W. K. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Yorita, A. M. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Tolosa, V. M. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)


    The enzyme immobilization process, one step in creating an enzymatic biosensor, was characterized and analyzed as a function of its physical properties. The neural glutamic biosensor is a flexible device, effectively minimizing trauma to the area of implantation. The Multielectrode Array (MEA) is composed primarily of a proprietary polymer which has been successfully implanted into human subjects in recent years. This polymer allows the device the pliability that other devices normally lack, though this poses some challenges to implantation. The electrodes are made of Platinum (Pt), and can range in number from eight to thirty two electrodes per device. These electrodes are electroplated with a semipermeable polymer layer to improve selectivity of the electrode to the neurotransmitter of interest, in this case glutamate. A signal is created from the interaction of glutamate in the brain with the glutamate oxidase (GluOx) which is immobilized on the surface of the electrode by using crosslinking chemistry in conjunction with glutaraldehyde and Bovine Serum Albumin (BSA). The glutamate is oxidized by glutamate oxidase, producing α-ketoglutarate and hydrogen peroxide (H2O2) as a by-product. The production of H2O2 is crucial for detection of the presence of the glutamate within the enzymatic coating, as it diffuses through the enzyme layer and oxidizes at the surface of the electrode. This oxidation is detectable by measurable change in the current using amperometry. Hence, the MEA allows for in vivo monitoring of neurotransmitter activity in real time. The sensitivity of the sensor to these neurotransmitters is dependent on the thickness of the layer, which is investigated in these experiments in order to optimize the efficacy of the device to detecting the substrate, once implanted.


    Directory of Open Access Journals (Sweden)



    Full Text Available The yield, productivity and cost for the enzymatic hydrolysis of cellulose to glucoseare crucial for the production of second generation ethanol. In the first study wehave evaluated the activity of several commercial cellulolytic enzymes and a crudeextract of a local strain of Trichoderma viride. The load used was 15 U ofcellulase/gram cellulose and 90 U of cellobiase/gram cellulose. The hydrolysis wascarried out at 50oC and pH 4,8 for 96 hours. The best cellulose hydrolysis yield of58% was obtained with the cocktail formed of crude cellulases from T. virideCMIT3.5 combined with Novozyme 188. This cocktail was used in the second study,when alkaline-steam pretreated wheat straw and corn stover where hydrolyzed at pH4,8 for 96 hours. The temperature was set at 50oC and 40oC. The hydrolysis at lowertemperature was tested for a future experiment of simultaneous hydrolysis andfermentation. An enzymatic assay using glucose-6-phosphate dehydrogenase wasused to determine exclusively glucose, instead of wide-range sugar DNS assay.Reporting to 100 grams of wet pretreated biomass, the following results wereobtained: 14.4 g% glucose for corn stover at 50oC and 13,0 g% at 40oC; 13,1 g%glucose for wheat straw at 50oC and 10.3 g% at 40oC. Considering that wheat strawcontain 36.6% glucose-based carbohydrates, the hydrolysis yields are between39.3% and 28.1%. Further studies, concerning the optimal parameters for cellulasecocktail will be made.


    Directory of Open Access Journals (Sweden)



    Full Text Available The yield, productivity and cost for the enzymatic hydrolysis of cellulose to glucose are crucial for the production of second generation ethanol. In the first study we have evaluated the activity of several commercial cellulolytic enzymes and a crude extract of a local strain of Trichoderma viride. The load used was 15 U of cellulase/gram cellulose and 90 U of cellobiase/gram cellulose. The hydrolysis was carried out at 50oC and pH 4,8 for 96 hours. The best cellulose hydrolysis yield of 58% was obtained with the cocktail formed of crude cellulases from T. viride CMIT3.5 combined with Novozyme 188. This cocktail was used in the second study, when alkaline-steam pretreated wheat straw and corn stover where hydrolyzed at pH 4,8 for 96 hours. The temperature was set at 50oC and 40oC. The hydrolysis at lower temperature was tested for a future experiment of simultaneous hydrolysis and fermentation. An enzymatic assay using glucose-6-phosphate dehydrogenase was used to determine exclusively glucose, instead of wide-range sugar DNS assay. Reporting to 100 grams of wet pretreated biomass, the following results were obtained: 14.4 g% glucose for corn stover at 50oC and 13,0 g% at 40oC; 13,1 g% glucose for wheat straw at 50oC and 10.3 g% at 40oC. Considering that wheat straw contain 36.6% glucose-based carbohydrates, the hydrolysis yields are between 39.3% and 28.1%. Further studies, concerning the optimal parameters for cellulase cocktail will be made.

  13. Maltodextrin-powered enzymatic fuel cell through a non-natural enzymatic pathway (United States)

    Zhu, Zhiguang; Wang, Yiran; Minteer, Shelley D.; Percival Zhang, Y.-H.

    Enzymatic fuel cells (EFCs) use a variety of fuels to generate electricity through oxidoreductase enzymes, such as oxidases or dehydrogenases, as catalysts on electrodes. We have developed a novel synthetic enzymatic pathway containing two free enzymes (maltodextrin phosphorylase and phosphoglucomutase) and one immobilized glucose-6-phosphate dehydrogenase that can utilize an oligomeric substrate maltodextrin for producing electrons mediated via a diaphorase and vitamin K 3 electron shuttle system. Three different enzyme immobilization approaches were compared based on electrostatic force entrapment, chemical cross-linking, and cross-linking with the aid of carbon nanotubes. At 10 mM glucose-6-phosphate (G6P) as a substrate concentration, the maximum power density of 0.06 mW cm -2 and retaining 42% of power output after 11 days were obtained through the method of chemical cross-linking with carbon nanotubes, approximately 6-fold and 3.5-fold better than those of the electrostatic force-based method, respectively. When changed to maltodextrin (degree of polymerization = 19) as the substrate, the EFC achieved a maximum power density of 0.085 mW cm -2. With the advantages of stable, low cost, high energy density, non-inhibitor to enzymes, and environmental friendly, maltodextrin is suggested to be an ideal fuel to power enzymatic fuel cells.

  14. Early life stress and serotonin transporter gene variation interact to affect the transcription of the glucocorticoid and mineralocorticoid receptors, and the co-chaperone FKBP5, in the adult rat brain.

    Directory of Open Access Journals (Sweden)

    Rick H. A. Van der Doelen


    Full Text Available The short allelic variant of the serotonin transporter (5-HTT promoter-linked polymorphic region (5-HTTLPR has been associated with the etiology of major depression by interaction with early life stress (ELS. A frequently observed endophenotype in depression is the abnormal regulation of levels of stress hormones such as glucocorticoids. It is hypothesized that altered central glucocorticoid influence on stress-related behavior and memory processes could underlie the depressogenic interaction of 5-HTTLPR and ELS. One possible mechanism could be the altered expression of the genes encoding the glucocorticoid and mineralocorticoid receptor (GR, MR and their inhibitory regulator FK506-binding protein 51 (FKBP5 in stress-related forebrain areas. To test this notion, we exposed heterozygous (5-HTT+/- and homozygous (5-HTT-/- serotonin transporter knockout rats and their wildtype littermates (5-HTT+/+ to daily 3 h maternal separations from postnatal day 2 to 14. In the medial prefrontal cortex (mPFC and hippocampus of the adult male offspring, we found that GR, MR and FKBP5 mRNA levels were affected by ELS x 5-HTT genotype interaction. Specifically, 5-HTT+/+ rats exposed to ELS showed decreased GR and FKBP5 mRNA in the dorsal and ventral mPFC, respectively. In contrast, 5-HTT+/- rats showed increased MR mRNA levels in the hippocampus and 5-HTT-/- rats showed increased FKBP5 mRNA in the ventral mPFC after ELS exposure. These findings indicate that 5-HTT genotype determines the specific adaptation of GR, MR and FKBP5 expression in response to early life adversity. Therefore, altered extra-hypothalamic glucocorticoid signaling should be considered to play a role in the depressogenic interaction of ELS and 5-HTTLPR.

  15. Oligoarginine peptides slow strand annealing and assist non-enzymatic RNA replication (United States)

    Jia, Tony Z.; Fahrenbach, Albert C.; Kamat, Neha P.; Adamala, Katarzyna P.; Szostak, Jack W.


    The non-enzymatic replication of RNA is thought to have been a critical process required for the origin of life. One unsolved difficulty with non-enzymatic RNA replication is that template-directed copying of RNA results in a double-stranded product. After strand separation, rapid strand reannealing outcompetes slow non-enzymatic template copying, which renders multiple rounds of RNA replication impossible. Here we show that oligoarginine peptides slow the annealing of complementary oligoribonucleotides by up to several thousand-fold; however, short primers and activated monomers can still bind to template strands, and template-directed primer extension can still occur, all within a phase-separated condensed state, or coacervate. Furthermore, we show that within this phase, partial template copying occurs even in the presence of full-length complementary strands. This method to enable further rounds of replication suggests one mechanism by which short non-coded peptides could have enhanced early cellular fitness, and potentially explains how longer coded peptides, that is, proteins, came to prominence in modern biology.

  16. Effect of harvest date on Arundo donax L. (giant reed) composition, ensilage performance, and enzymatic digestibility. (United States)

    Liu, Shan; Ge, Xumeng; Liu, Zhe; Li, Yebo


    Composition and ensilage performance of giant reed harvested in August, October, November, and December, were evaluated and compared. Generally, late-harvested giant reed had higher dry matter content, lower nitrogen content, and higher water soluble carbohydrates (WSC) content than early-harvested giant reed. During 90days of ensilage, giant reed harvested in October, November, and December showed dry matter losses of about 1%, while giant reed harvested in August showed a higher dry matter loss of about 8%. During the ensilage process, more lactic acid was produced in late-harvested giant reed than in early-harvested giant reed. Late-harvested giant reed had a higher lignin content and lower enzymatic digestibility than early-harvested giant reed. However, enzymatic digestibility of all the giant reed biomass was improved by the 90-day ensilage process, reaching levels of 43-46%. In summary, ensilage could be used for storing giant reed biomass harvested at different times and for improving its digestibility.

  17. Non-enzymatic modification of aminophospholipids by carbonyl-amine reactions. (United States)

    Naudí, Alba; Jové, Mariona; Ayala, Victòria; Cabré, Rosanna; Portero-Otín, Manuel; Pamplona, Reinald


    Non-enzymatic modification of aminophospholipids by lipid peroxidation-derived aldehydes and reducing sugars through carbonyl-amine reactions are thought to contribute to the age-related deterioration of cellular membranes and to the pathogenesis of diabetic complications. Much evidence demonstrates the modification of aminophospholipids by glycation, glycoxidation and lipoxidation reactions. Therefore, a number of early and advanced Maillard reaction-lipid products have been detected and quantified in different biological membranes. These modifications may be accumulated during aging and diabetes, introducing changes in cell membrane physico-chemical and biological properties.

  18. Affective Urbanism

    DEFF Research Database (Denmark)

    Samson, Kristine

    Urban design and architecture are increasingly used as material and affective strategies for setting the scene, for manipulation and the production of urban life: The orchestration of atmospheres, the framing and staging of urban actions, the programming for contemplation, involvement, play, expe...... affects can be choreographed and designed intentionally or whether it arises from unpredictable circumstances within urbanity itself....

  19. Affective Maps

    DEFF Research Database (Denmark)

    Salovaara-Moring, Inka

    of environmental knowledge production. It uses InfoAmazonia, the databased platform on Amazon rainforests, as an example of affective geo-visualization within information mapping that enhances embodiment in the experience of the information. Amazonia is defined as a digitally created affective (map)space within...

  20. Enzymatic production of biodiesel from canola oil using immobilized lipase

    Energy Technology Data Exchange (ETDEWEB)

    Dizge, Nadir; Keskinler, Buelent [Department of Environmental Engineering, Gebze Institute of Technology, Gebze 41400 (Turkey)


    In the present work, a novel method for immobilization of lipase within hydrophilic polyurethane foams using polyglutaraldehyde was developed for the immobilization of Thermomyces lanuginosus lipase to produce biodiesel with canola oil and methanol. The enzyme optimum conditions were not affected by immobilization and the optimum pH for free and immobilized enzyme were 6, resulting in 80% immobilization yield. Using the immobilized lipase T. lanuginosus, the effects of enzyme loading, oil/alcohol molar ratio, water concentration, and temperature in the transesterification reaction were investigated. The optimal conditions for processing 20 g of refined canola oil were: 430 {mu}g lipase, 1:6 oil/methanol molar ratio, 0.1 g water and 40 C for the reactions with methanol. Maximum methyl esters yield was 90% of which enzymatic activity remained after 10 batches, when tert-butanol was adopted to remove by-product glycerol during repeated use of the lipase. The immobilized lipase proved to be stable and lost little activity when was subjected to repeated uses. (author)

  1. Early osmotic, antioxidant, ionic, and redox responses to salinity in leaves and roots of Indian mustard (Brassica juncea L.). (United States)

    Ranjit, Singh Laxmi; Manish, Pandey; Penna, Suprasanna


    Salt-stress-induced alterations in osmotic, ionic, and redox responses were studied in the early period of treatment (30 min to 5 days) in seedlings of Brassica juncea L. Roots and shoots under mild (50 mM) and severe (250 mM) NaCl stress were analyzed for growth, oxidative stress, osmolyte accumulation, antioxidant defense, and redox state. Growth reduction was less pronounced in the early time period of salt stress while oxidative damage increased linearly and in a sustained manner under severe stress up to 6 h. An early and transient reactive oxygen species (ROS) burst, as evidenced by superoxide and hydrogen peroxide level was observed, followed by activation of enzymatic antioxidant system (GPX, SOD, CAT, and GR) in both root and shoot. The enzymatic activity was not affected much under mild stress particularly at early phase; however, severe stress induced a significant increase in the activity of antioxidant enzymes. Root ascorbate was progressively accumulated, and its redox state maintained in the early time phase of treatment under mild stress while increase in root and shoot glutathione content was recorded under mild stress at 5 days when the active ascorbate pool decreased. While early period of salt stress showed significant Na(+) accumulation over control, plants subjected to mild stress measured less Na(+) accumulation up to 5 days compared to severely stressed plants. The results showed an early induction of differential responses to salt stress in roots and shoots of Brassica which include growth limitations, reduced relative water content, increased osmolytes, redox state, and antioxidant system, and a significant Na(+) increase. The results also indicate that roots and shoots may have distinct mechanisms of responses to salt stress.

  2. Unraveling the Enzymatic Basis of Wine “Flavorome”: A Phylo-Functional Study of Wine Related Yeast Species


    Ignacio eBelda; Javier eRuiz; Ana eAlastruey-Izquierdo; Eva eNavascués; Domingo eMarquina; Antonio eSantos


    Non-Saccharomyces yeasts are a heterogeneous microbial group involved in the early stages of wine fermentation. The high enzymatic potential of these yeasts makes them a useful tool for increasing the final organoleptic characteristics of wines in spite of their low fermentative power. Their physiology and contribution to wine quality are still poorly understood, with most current knowledge being acquired empirically and in most cases based in single species and strains. This work analyzed th...

  3. The influence of selected hypotensive drugs on the bioavailability of minerals from buckwheat groats in vitro enzymatic digestion

    Directory of Open Access Journals (Sweden)

    Joanna Suliburska


    Full Text Available Background. The mineral status in hypertensive patients may be affected by hypotensive drugs. The aim of this study was to estimate the influence of hypotensive drugs (angiotensine converting enzyme inhibitors (ACE-I, b-blockers, Ca-antagonists, diuretics on the potential bioavailability of magnesium, iron, zinc and copper from buckwheat groats in vitro enzymatic digestion. Material and methods. The degree of release of magnesium, iron, zinc and copper from buckwheat groats was determined with and without (the control sample an addition of hypotensive drugs. Four antihypertensive drugs in one dose (one tablet per sample were analysed: metocard (a β-blocker, cardilopin (a Ca-antagonist, apo-perindox (ACE-I and indapen (a diuretic. The samples were subjected to enzymatic digestion under in vitro conditions. The content of minerals in buckwheat groats before and after enzymatic digestion was determined by flame atomic absorption spectrometry (AAS. Results. It was found that cardilopin (amlodipine and indapen (indapamide significantly increased the release of zinc from groats. The degree of release of magnesium was higher and the release of iron was lower in samples with apo-perindox (perindopril than in the control group. The release of copper was significantly decreased by indapen (indapamid. Conclusions. Amlodipine, perindopril and indapamide affected the release of magnesium, iron, zinc and copper from buckwheat groats in vitro enzymatic digestion.

  4. Investigation on chilling affected banana plantations during early spring season and restoration strategies%2011年早春我国香蕉寒害调查及寒害后恢复对策

    Institute of Scientific and Technical Information of China (English)

    林贵美; 李小泉; 韦绍龙; 邹瑜; 牟海飞; 李朝生; 韦华芳; 张进忠


    [目的]了解2011年早春低温对我国香蕉生产的影响,提出寒害后香蕉恢复生产的技术措施,为香蕉抗寒栽培提供技术支持.[方法]2011年3月9~16日对我国香蕉主产区广西、广东、海南、云南、福建进行分片区实地寒害调查,对挂果株及待生产用的大、中、小吸芽进行田间解剖分析诊断.[结果]挂果株受寒害影响较吸芽植株敏感,大、中、小吸芽抗寒能力依次增强;同纬度区域高海拔香蕉受寒害比低海拔严重;此次低温对广西中部区域香蕉影响最为严重,挂果株产量损失超过50%,且只能留中小型吸芽作为生产株;广东、海南区域香蕉受寒害影响最小,其他区域居中.[建议]采用覆盖天地膜过冬、喷水保湿、套袋护果御寒等措施;选留受寒害后萌发的吸芽为下造生产株,做好蕉园管理工作,促进吸芽早萌发、早定芽;选育抗寒品种.%[ Objective ]The experiment was conducted to elucidate the effects of low temperature on banana yield during early spring season of the year 2011 in order to develop proper measures for recovering banana plantations after being injured, and to sustain banana production under cold temperature conditions. [Method]Investigations were carried out to observe the effects of chilling temperatures on banana plantations in main banana production regions viz., Guangxi, Guangdong, Hainan, Yunnan, and Fujian from March 9th to March 16th, 2011. Large, middle and small sized banana suckers and fruited plants were dissected and diagnosed under field conditions. [Result]The results showed that the fruited banana plants were more sensitive to low temperature compared to suckers. Large sized suckers responded more imperceptibly in cold weather compared to the smaller ones. The chilling damages to banana plants were more severe at high altitude compared to low altitude at same latitude. Banana plantations in central areas of Guangxi were severly affected

  5. Responses of enzymatic antioxidants and non-enzymatic antioxidants in the cyanobacterium Microcystis aeruginosa to the allelochemical ethyl 2-methyl acetoacetate (EMA) isolated from reed (Phragmites communis). (United States)

    Hong, Yu; Hu, Hong-Ying; Xie, Xing; Li, Feng-Min


    Macrophytic allelochemicals are considered an environment-friendly and promising alternative to control algal bloom. However, studies examining the potential mechanisms of inhibitory allelochemicals on algae are few. The allelochemical ethyl 2-methyl acetoacetate (EMA), isolated from reed (Phragmites communis), was a strong allelopathic inhibitor on the growth of Microcystis aeruginosa. EMA-induced antioxidant responses were investigated in the cyanobacterium M. aeruginosa to understand the mechanism of EMA inhibition on algal growth. The activities of enzymatic antioxidants superoxide dismutase (SOD) and catalase (CAT), and the contents of non-enzymatic antioxidants reduced glutathione (GSH) and ascorbic acid (AsA) of M. aeruginosa cells were analyzed after treatments with different concentrations of EMA. Exposure of M. aeruginosa to EMA caused changes in enzyme activities and contents of non-enzymatic antioxidants in different manners. The decrease in SOD activity occurred first after 4 h of EMA exposure, and more markedly after 40 h. CAT activity did not change after 4 h of EMA exposure, but increased obviously after 40 h. The contents of AsA and GSH were increased greatly by EMA after 4 h. After 60 h, low EMA concentrations still increased the CAT activity and the contents of AsA and GSH, but high EMA concentrations started to impose a marked suppression on them. EMA increased dehydroascorbate (DHAsA) and oxidized glutathione (GSSG) contents during all exposure times. After 60 h, the regeneration rates of AsA and GSH (represented by the AsA/DHAsA ratio and GSH/GSSG ratio, respectively) were reduced by high EMA concentrations. These results suggest that the activation of CAT and the availability of AsA and GSH at early exposure are important to counteract the oxidative stress induced by EMA, and the inactivation of SOD may be crucial to the growth inhibition of M. aeruginosa by EMA.

  6. Enzymatic digestion of articular cartilage results in viscoelasticity changes that are consistent with polymer dynamics mechanisms

    Directory of Open Access Journals (Sweden)

    June Ronald K


    Full Text Available Abstract Background Cartilage degeneration via osteoarthritis affects millions of elderly people worldwide, yet the specific contributions of matrix biopolymers toward cartilage viscoelastic properties remain unknown despite 30 years of research. Polymer dynamics theory may enable such an understanding, and predicts that cartilage stress-relaxation will proceed faster when the average polymer length is shortened. Methods This study tested whether the predictions of polymer dynamics were consistent with changes in cartilage mechanics caused by enzymatic digestion of specific cartilage extracellular matrix molecules. Bovine calf cartilage explants were cultured overnight before being immersed in type IV collagenase, bacterial hyaluronidase, or control solutions. Stress-relaxation and cyclical loading tests were performed after 0, 1, and 2 days of incubation. Results Stress-relaxation proceeded faster following enzymatic digestion by collagenase and bacterial hyaluronidase after 1 day of incubation (both p ≤ 0.01. The storage and loss moduli at frequencies of 1 Hz and above were smaller after 1 day of digestion by collagenase and bacterial hyaluronidase (all p ≤ 0.02. Conclusion These results demonstrate that enzymatic digestion alters cartilage viscoelastic properties in a manner consistent with polymer dynamics mechanisms. Future studies may expand the use of polymer dynamics as a microstructural model for understanding the contributions of specific matrix molecules toward tissue-level viscoelastic properties.


    Directory of Open Access Journals (Sweden)



    Full Text Available Robust immobilization techniques that preserve the activity of biomolecules havemany potential applications. In recent years, a number of new bioimobilisationmethods in sol-gel-derived materials were reported. The interactions between thebiomolecule and the inorganic material determine the degree to which thebiomolecule retains its native properties. The newer technological developments inthe field of immobilized biocatalysts can offer the possibility of a wider and moreeconomical exploitation of biocatalysts in biological applications, food and feedindustry, medicine, and in the development of bioprocess monitoring devices, like thebiosensors.The aim of this study was to obtain immobilized enzymatic preparations by methodswhich affect enzyme conformations and kinetic parameters as less as possible. Weimmobilized the enzymatic preparation with protease activity produced by a Bacilluslicheniformis B 40 local strain by physical bonding on ceramics and entrapment intosol-gel-derived glasses obtained from tetraethyl orthosilicate (TEOS, deposited inthin layer on a ceramic support (entrapment/deposition. Both physically adsorbedand entrapped/deposited enzymes follow Michaelis-Menten kinetics, similar with thesoluble enzyme. In the case of immobilized enzymes, the apparent Michaelisconstant, Km, was greater than that of the native one, as it was expected. The kineticparameters indicate that the enzymatic preparations adsorbed on ceramic supportand entrapped/deposited show less affinity for the substrate, Km being 1.3 and 2.1times higher than that of the native enzyme, respectively. The maximum velocityincreased also by 3.5 and 7.9 times respectively, compared with the free counterpart(according to Lineweaver-Burk linearization.

  8. Chemically-enzymatic synthesis of photosensitive DNA. (United States)

    Westphal, Kinga; Zdrowowicz, Magdalena; Zylicz-Stachula, Agnieszka; Rak, Janusz


    The sensitizing propensity of radio-/photosensitizing nucleoside depends on DNA sequence surrounding a sensitizer. Therefore, in order to compare sensitizers with regard to their ability to induce a DNA damage one has to study the sequence dependence of damage yield. However, chemical synthesis of oligonucleotides labeled with sensitizing nucleosides is hindered due to the fact that a limited number of such nucleoside phosphoramidites are accessible. Here, we report on a chemically-enzymatic method, employing a DNA polymerase and ligase, that enables a modified nucleoside, in the form of its 5'-triphosphate, to be incorporated into DNA fragment in a pre-determined site. Using such a protocol two double-stranded DNA fragments - a long one, 75 base pairs (bp), and a short one, 30bp in length - were pin-point labeled with 5-bromodeoxyuridine. Four DNA polymerases together with DHPLC for the inspection of reaction progress were used to optimize the process under consideration. As an ultimate test showing that the product possessing an assumed nucleotide sequence was actually obtained, we irradiated the synthesized oligonucleotide with UVB photons and analyzed its photoreactivity with the LC-MS method. Our results prove that a general approach enabling precise labeling of DNA with any nucleoside modification processed by DNA polymerase and ligase has been worked out.

  9. Aqueous enzymatic extraction of Moringa oleifera oil. (United States)

    Mat Yusoff, Masni; Gordon, Michael H; Ezeh, Onyinye; Niranjan, Keshavan


    This paper reports on the extraction of Moringa oleifera (MO) oil by using aqueous enzymatic extraction (AEE) method. The effect of different process parameters on the oil recovery was discovered by using statistical optimization, besides the effect of selected parameters on the formation of its oil-in-water cream emulsions. Within the pre-determined ranges, the use of pH 4.5, moisture/kernel ratio of 8:1 (w/w), and 300stroke/min shaking speed at 40°C for 1h incubation time resulted in highest oil recovery of approximately 70% (goil/g solvent-extracted oil). These optimized parameters also result in a very thin emulsion layer, indicating minute amount of emulsion formed. Zero oil recovery with thick emulsion were observed when the used aqueous phase was re-utilized for another AEE process. The findings suggest that the critical selection of AEE parameters is key to high oil recovery with minimum emulsion formation thereby lowering the load on the de-emulsification step.

  10. Effects of thermal and enzymatic treatments and harvesting time on the microbial quality and chemical composition of fibre hemp (Cannabis sativa L.)

    DEFF Research Database (Denmark)

    Nykter, M.; Kymalainen, H.R.; Thomsen, Anne Belinda


    harvested after stand retting in the field before frost, after early frost or in the following spring. The enzymatic treatment of hemp promoted growth of moulds on the fibres (500-fold increase in colony-forming units (cfu)), whereas steam explosion reduced the amount of moulds to a relatively constant...

  11. Effect of enzymatic mash treatment and storage on phenolic composition, antioxidant activity, and turbidity of cloudy apple juice. (United States)

    Oszmiański, Jan; Wojdylo, Aneta; Kolniak, Joanna


    The effects of different commercial enzymatic mash treatments on yield, turbidity, color, and polyphenolic and sediment of procyanidins content of cloudy apple juice were studied. Addition of pectolytic enzymes to mash treatment had positive effect on the production of cloud apple juices by improving polyphenolic contents, especially procyanidins and juice yields (68.3% in control samples to 77% after Pectinex Yield Mash). As summary of the effect of enzymatic mash treatment, polyphenol contents in cloudy apple juices significantly increased after Pectinex Yield Mash, Pectinex Smash XXL, and Pectinex XXL maceration were applied but no effect was observed after Pectinex Ultra-SPL I Panzym XXL use, compared to the control samples. The content of polymeric procyanidins represented 50-70% of total polyphenols, but in the present study, polymeric procyanidins were significantly lower in juices than in fruits and also affected by enzymatic treatment (Pectinex AFP L-4 and Panzym Yield Mash) compared to the control samples. The enzymatic treatment decreased procyanidin content in most sediment with the exception of Pectinex Smash XXL and Pectinex AFP L-4. Generally in samples that were treated by pectinase, radical scavenging activity of cloudy apple juices was increased compared to the untreated reference samples. The highest radical scavenging activity was associated with Pectinex Yield Mash, Pectinex Smash XXL, and Pectinex XXL enzyme and the lowest activity with Pectinex Ultra SP-L and Pectinex APFL-4. However, in the case of enzymatic mash treatment cloudy apple juices showed instability of turbidity and low viscosity. These results must be ascribed to the much higher hydrolysis of pectin by enzymatic preparation which is responsible for viscosity. During 6 months of storage at 4 degrees C small changes in analyzed parameters of apple juices were observed.

  12. Recent insights in enzymatic synthesis of fructooligosaccharides from inulin. (United States)

    Singh, Ram Sarup; Singh, Rupinder Pal; Kennedy, John F


    In the past few years, people are paying more attention to their dietary habits, and functional foods are playing a key role in maintaining the health of man. Prebiotics are considered as a main component of the functional foods which are usually composed of short chains of carbohydrates. Fructooligosaccharides (FOSs) are considered as one of the main group of prebiotics which have recognisable bifidogenic properties. FOSs are obtained either by extraction from various plant materials or by enzymatic synthesis from different substrates. Enzymatically, these can be obtained either from sucrose using fructosyltransferase or from inulin by endoinulinase. Inulin is a potent substrate for the enzymatic production of FOSs. This review article will provide an overview on the inulin as potent substrate, microbial sources of endoinulinases, enzymatic synthesis of FOSs from inulin, commercial status of FOSs, and their future perspectives.

  13. Process technology for multi-enzymatic reaction systems

    DEFF Research Database (Denmark)

    Xue, Rui; Woodley, John M.


    synthesis and fermentation as an alternative to chemical-catalysis for the production of pharmaceuticals and fine chemicals. In particular, the use of multiple enzymes is of special interest. However, many challenges remain in the scale-up of a multi-enzymatic system. This review summarizes and discusses...... the technology options and strategies that are available for the development of multi-enzymatic processes. Some engineering tools, including kinetic models and operating windows, for developing and evaluating such processes are also introduced....

  14. Differential cDNA cloning by enzymatic degrading subtraction (EDS).



    We describe a new method, called enzymatic degrading subtraction (EDS), for the construction of subtractive libraries from PCR amplified cDNA. The novel features of this method are that i) the tester DNA is blocked by thionucleotide incorporation; ii) the rate of hybridization is accelerated by phenol-emulsion reassociation; and iii) the driver cDNA and hybrid molecules are enzymatically removed by digestion with exonucleases III and VII rather than by physical partitioning. We demonstrate th...

  15. Enzymatically induced motion at nano- and micro-scales (United States)

    Gáspár, Szilveszter


    In contrast to adenosine triphosphate (ATP)-dependent motor enzymes, other enzymes are little-known as ``motors'' or ``pumps'', that is, for their ability to induce motion. The enhanced diffusive movement of enzyme molecules, the self-propulsion of enzyme-based nanomotors, and liquid pumping with enzymatic micropumps were indeed only recently reported. Enzymatically induced motion can be achieved in mild conditions and without the use of external fields. It is thus better suited for use in living systems (from single-cell to whole-body) than most other ways to achieve motion at small scales. Enzymatically induced motion is thus not only new but also important. Therefore, the present work reviews the most significant discoveries in enzymatically induced motion. As we will learn, freely diffusing enzymes enhance their diffusive movement by nonreciprocal conformational changes which parallel their catalytic cycles. Meanwhile, enzyme-modified nano- and micro-objects turn chemical energy into kinetic energy through mechanisms such as bubble recoil propulsion, self-electrophoresis, and self-diffusiophoresis. Enzymatically induced motion of small objects ranges from enhanced diffusive movement to directed motion at speeds as high as 1 cm s-1. In spite of the progress made in understanding how the energy of enzyme reactions is turned into motion, most enzymatically powered devices remain inefficient and need improvements before we will witness their application in real world environments.

  16. Enhanced enzymatic conversion with freeze pretreatment of rice straw

    Energy Technology Data Exchange (ETDEWEB)

    Chang, Ken-Lin; Thitikorn-amorn, Jitladda; Ou, Bay-Ming; Chen, Shan-He; Huang, Po-Jung [Institute of Biological Chemistry and Genomics Research Center Academia Sinica, Nankang, Taipei 115 (China); Hsieh, Jung-Feng [Department of Food Science, Fu Jen Catholic University, Xin Zhuang, Taipei 242 (China); Ratanakhanokchai, Khanok [School of Bioresources and Technology, King Mongkut' s University of Technology Thonburi, Bangkok 10150 (Thailand); Chen, Shui-Tein [Institute of Biological Chemistry and Genomics Research Center Academia Sinica, Nankang, Taipei 115 (China); Institute of Biochemical Sciences, College of Life Science, National Taiwan University, Taipei 106 (China)


    Production of bioethanol by the conversion of lignocellulosic waste has attracted much interest in recent years, because of its low cost and great potential availability. The pretreatment process is important for increasing the enzymatic digestibility of lignocellulosic materials. Enzymatic conversion with freeze pretreatment of rice straw was evaluated in this study. The freeze pretreatment was found to significantly increase the enzyme digestibility of rice straw from 48% to 84%. According to the results, enzymatic hydrolysis of unpretreated rice straw with 150 U cellulase and 100 U xylanase for 48 h yielded 226.77 g kg{sup -1} and 93.84 g kg{sup -1} substrate-reducing sugars respectively. However, the reducing sugar yields from freeze pretreatment under the same conditions were 417.27 g kg{sup -1} and 138.77 g kg{sup -1} substrate, respectively. In addition, hydrolyzates analysis showed that the highest glucose yield obtained during the enzymatic hydrolysis step in the present study was 371.91 g kg{sup -1} of dry rice straw, following pretreatment. Therefore, the enhanced enzymatic conversion with freeze pretreatment of rice straw was observed in this study. This indicated that freeze pretreatment was highly effective for enzymatic hydrolysis and low environmental impact. (author)

  17. Kinetics of enzymatic hydrolysis of methyl ricinoleate

    Directory of Open Access Journals (Sweden)

    Neeharika, T. S.V.R.


    Full Text Available Ricinoleic acid is an unsaturated hydroxy fatty acid that naturally occurs in castor oil in proportions of up to 85–90%. Ricinoleic acid is a potential raw material and finds several applications in coatings, lubricant formulations and pharmaceutical areas. Enzymatic hydrolysis of castor oil is preferred over conventional hydrolysis for the preparation of ricinoleic acid to avoid estolide formation. A kinetics analysis of the enzymatic hydrolysis of Methyl Ricinoleate in the presence of Candida antarctica Lipase B was carried out in this study by varying reaction temperature (40–60 °C and enzyme concentration (2–5%. The optimal conditions were found to be 6 h reaction time, temperature 60°C, buffer to methyl ricinoleate ratio 2:1(v/w and 4% enzyme concentration to achieve a maximum conversion of 98.5%. A first order reversible reaction kinetic model was proposed to describe this reaction and a good agreement was observed between the experimental data and the model values. The effect of temperature on the forward reaction rate constant was determined by fitting data to the Arrhenius equation. The activation energy for forward reaction was found to be 14.69 KJ·mol−1.El ácido ricinoleico es un hidroxiácido insaturado que se produce naturalmente en el aceite de ricino en proporciones de hasta el 85–90%. El ácido ricinoleico es una materia prima con gran potencial y tiene aplicaciones en revestimientos, formulaciones lubricantes y en áreas farmacéuticas. Para la preparación del ácido ricinoleico se prefiere la hidrólisis enzimática del aceite de ricino a la hidrólisis convencional, para evitar la formación de estólidos. En este estudio se llevó a cabo la cinética de la hidrólisis enzimática del ricinoleato de metilo en presencia de lipasa de Candida antarctica B mediante la variación de la temperatura de reacción (40–60 °C y la concentración de la enzima (2–5%. Las condiciones óptimas de la reacción para

  18. Effects of cadmium on enzymatic and non-enzymatic antioxidative defences of rice (Oryza sativa L.). (United States)

    Yu, Fangming; Liu, Kehui; Li, Mingshun; Zhou, Zhenming; Deng, Hua; Chen, Bin


    The effects of 60-d cadmium (Cd) exposure on enzymatic and non-enzymatic antioxidative system of Oryza sativa L. seedlings at tillering stage were studied using soil culture experiment. Research findings showed that chlorophyll content of Oryza sativa L. declined with the increase in soil metal concentration. Cd pollution induced the antioxidant stress by inducing O2(-1) and H2O2, which increased in plants; at the same time, MDA as the final product of peroxidation of membrane lipids, accumulated in plant. The antioxidant enzyme system was initiated under the Cd exposure, i.e. almost all the activities of superoxide dismutase (SOD), peroxidase, catalase, glutathione peroxidase, and ascorbate peroxidase were elevated both in leaves and roots. The non-protein thiols including phytochelatins and glutathione to scavenge toxic free radicals caused by Cd stress was also studied. The contents of phytochelatins and glutathione were about 3.12-6.65-fold and 3.27-10.73-fold in leaves, against control; and the corresponding values were about 3.53-9.37-fold and 1.41-5.11-fold in roots, accordingly.

  19. Enzymatic Catalysis at Interfaces—Heterophase Systems as Substrates for Enzymatic Action

    Directory of Open Access Journals (Sweden)

    Katharina Landfester


    Full Text Available Several important enzymatic reactions occurring in nature, such as, e.g., the digestion of fat, proceed only at the interface of two immiscible phases. Typically, these systems consist of an organic substrate, dispersed in an aqueous continuous phase, with a specialized enzyme capable of working at the interface. For adopting such a system for organic synthesis, a stable heterophase system with a large interfacial area is required. These prerequisites can be found in so-called miniemulsions. Such liquid-liquid heterophase systems feature droplets with sizes smaller than 500 nm, and more importantly, these emulsions do not suffer from Ostwald ripening, as conventional emulsions do. Consequently, the droplets show long-term stability, even throughout reactions conducted in the droplets. In this review, we will briefly discuss the physicochemical background of miniemulsions, provide a comprehensive overview of the enzymatically catalyzed reactions conducted in miniemulsions and, as data are available, to compare the most important features to conventional systems, as reverse microemulsions, (macroemulsions and solvent-based systems.

  20. Nanocrystal Bioassembly: Asymmetry, Proximity, and Enzymatic Manipulation

    Energy Technology Data Exchange (ETDEWEB)

    Claridge, Shelley A. [Univ. of California, Berkeley, CA (United States)


    Research at the interface between biomolecules and inorganic nanocrystals has resulted in a great number of new discoveries. In part this arises from the synergistic duality of the system: biomolecules may act as self-assembly agents for organizing inorganic nanocrystals into functional materials; alternatively, nanocrystals may act as microscopic or spectroscopic labels for elucidating the behavior of complex biomolecular systems. However, success in either of these functions relies heavily uponthe ability to control the conjugation and assembly processes.In the work presented here, we first design a branched DNA scaffold which allows hybridization of DNA-nanocrystal monoconjugates to form discrete assemblies. Importantly, the asymmetry of the branched scaffold allows the formation of asymmetric2assemblies of nanocrystals. In the context of a self-assembled device, this can be considered a step toward the ability to engineer functionally distinct inputs and outputs.Next we develop an anion-exchange high performance liquid chromatography purification method which allows large gold nanocrystals attached to single strands of very short DNA to be purified. When two such complementary conjugates are hybridized, the large nanocrystals are brought into close proximity, allowing their plasmon resonances to couple. Such plasmon-coupled constructs are of interest both as optical interconnects for nanoscale devices and as `plasmon ruler? biomolecular probes.We then present an enzymatic ligation strategy for creating multi-nanoparticle building blocks for self-assembly. In constructing a nanoscale device, such a strategy would allow pre-assembly and purification of components; these constructs can also act as multi-label probes of single-stranded DNA conformational dynamics. Finally we demonstrate a simple proof-of-concept of a nanoparticle analog of the polymerase chain reaction.

  1. Chemicals effect on the enzymatic digestibility of rape straw over the thermo-mechanical pretreatment using a continuous twin screw-driven reactor (CTSR). (United States)

    Um, Byung-Hwan; Choi, Chang Ho; Oh, Kyeong Keun


    Rape straw pretreated by a continuous twin screw-driven reactor (CTSR) with hot water presented a distinctive particle-size distribution profile as a function of the operating temperature. The relative amount of finer particle size dramatically increased as the ratio of solid to liquid was increased. Size reduction through physical CTSR process effectively promoted the enzymatic hydrolysis of pretreated rape straw. Meanwhile, the crystallinity of the physically pretreated straw was not a greater factor affecting the enzyme digestibility. The glucose conversion from the enzymatic hydrolysis of the straw pretreated by CTSR with hot water was maximized at 52%. Using the chemicals as catalyst have affected considerably for increasing the digestibility at same condition with hot water pretreatment. The enzymatic digestibilities of the straw pretreated by CTSR with sodium hydroxide and sulfuric acid were 60% and 77%, respectively.

  2. Affect Regulation

    DEFF Research Database (Denmark)

    Pedersen, Signe Holm; Poulsen, Stig Bernt; Lunn, Susanne


    Gergely and colleagues’ state that their Social Biofeedback Theory of Parental Affect Mirroring” can be seen as a kind of operationalization of the classical psychoanalytic concepts of holding, containing and mirroring. This article examines to what extent the social biofeedback theory of parenta...

  3. Enzymatic and non-enzymatic antioxidant potentials of Chlorella vulgaris grown in effluent of a confectionery industry



    Enzymatic and non-enzymatic antioxidant potentials of Chlorella vulgaris have gained considerable importance in recent decades. C. vulgaris strain highly tolerant to extreme pH variations was isolated and mass-cultivated in the wastewater from a confectionery industry. C.vulgaris showed better growth in wastewater than in improvised CFTRI medium. The microalgal biomass was then screened for the following antioxidants: peroxidase, superoxide dismutase, polyphenol oxidase, glutathione peroxidas...

  4. Actin cytoskeleton contributes to the elastic modulus of embryonic tendon during early development. (United States)

    Schiele, Nathan R; von Flotow, Friedrich; Tochka, Zachary L; Hockaday, Laura A; Marturano, Joseph E; Thibodeau, Jeffrey J; Kuo, Catherine K


    Tendon injuries are common and heal poorly. Strategies to regenerate or replace injured tendons are challenged by an incomplete understanding of normal tendon development. Our previous study showed that embryonic tendon elastic modulus increases as a function of developmental stage. Inhibition of enzymatic collagen crosslink formation abrogated increases in tendon elastic modulus at late developmental stages, but did not affect increases in elastic modulus of early stage embryonic tendons. Here, we aimed to identify potential contributors to the mechanical properties of these early stage embryonic tendons. We characterized tendon progenitor cells in early stage embryonic tendons, and the influence of actin cytoskeleton disruption on tissue elastic modulus. Cells were closely packed in embryonic tendons, and did not change in density during early development. We observed an organized network of actin filaments that seemed contiguous between adjacent cells. The actin filaments exhibited a crimp pattern with a period and amplitude that matched the crimp of collagen fibers at each developmental stage. Chemical disruption of the actin cytoskeleton decreased tendon tissue elastic modulus, measured by atomic force microscopy. Our results demonstrate that early developmental stage embryonic tendons possess a well organized actin cytoskeleton network that contributes significantly to tendon tissue mechanical properties.

  5. Optimizing Wound Bed Preparation With Collagenase Enzymatic Debridement (United States)

    McCallon, Stanley K.; Weir, Dorothy; Lantis, John C.


    Difficult-to-heal and chronic wounds affect tens of millions of people worldwide. In the U.S. alone, the direct cost for their treatment exceeds $25 billion. Yet despite advances in wound research and treatment that have markedly improved patient care, wound healing is often delayed for weeks or months. For venous and diabetic ulcers, complete wound closure is achieved in as few as 25%–50% of chronic or hard-to-heal wounds. Wound bed preparation and the consistent application of appropriate and effective debridement techniques are recommended for the optimized treatment of chronic wounds. The TIME paradigm (Tissue, Inflammation/infection, Moisture balance and Edge of wound) provides a model to remove barriers to healing and optimize the healing process. While we often think of debridement as an episodic event that occurs in specific care giver/patient interface. There is the possibility of a maintenance debridement in which the chronic application of a medication can assist in both the macroscopic and microscopic debridement of a wound. We review the various debridement therapies available to clinicians in the United States, and explore the characteristics and capabilities of clostridial collagenase ointment (CCO), a type of enzymatic debridement, that potentially allows for epithelialization while debriding. It appears that in the case of CCO it may exert this influences by removal of the necrotic plug while promoting granulation and sustaining epithelialization. It is also easily combined with other methods of debridement, is selective to necrotic tissue, and has been safely used in various populations. We review the body of evidence has indicated that this concept of maintenance debridement, especially when combined episodic debridement may add a cost an efficacious, safe and cost-effective choice for debridement of cutaneous ulcers and burn wounds and it will likely play an expanding role in all phases of wound bed preparation. PMID:26442207

  6. Affective Networks

    Directory of Open Access Journals (Sweden)

    Jodi Dean


    Full Text Available This article sets out the idea of affective networks as a constitutive feature of communicative capitalism. It explores the circulation of intensities in contemporary information and communication networks, arguing that this circulation should be theorized in terms of the psychoanalytic notion of the drive. The article includes critical engagements with theorists such as Guy Debord, Jacques Lacan, Tiziana Terranova, and Slavoj Zizek.

  7. Early luteal phase endocrine profile is affected by the mode of triggering final oocyte maturation and the luteal phase support used in recombinant follicle-stimulating hormone-gonadotropin-releasing hormone antagonist in vitro fertilization cycles

    DEFF Research Database (Denmark)

    Fatemi, Human M; Polyzos, Nikolaos P; van Vaerenbergh, Inge;


    To assess endocrine differences during early luteal phase according to mode of triggering final oocyte maturation with or without luteal phase support (LPS).......To assess endocrine differences during early luteal phase according to mode of triggering final oocyte maturation with or without luteal phase support (LPS)....

  8. Early Childhood Special Music Education (United States)

    Darrow, Alice-Ann


    The process of early intervention is a critical component of Early Childhood Special Music Education. Early intervention is the process of providing services, education, and support to young children who have disabilities or to children who are at-risk of developing needs that may affect their physical, cognitive, or emotional development. The…

  9. Enzymatic accessibility of fiber hemp is enhanced by enzymatic or chemical removal of pectin. (United States)

    Pakarinen, A; Zhang, J; Brock, T; Maijala, P; Viikari, L


    Pectinolytic enzymes, steam explosion and alkaline treatment were used to assess the role of pectin for the accessibility of hydrolytic enzymes in the enzymatic hydrolysis of biomass. Hemp (Cannabis sativa L.), a potential energy crop especially in boreal climate with a low need of fertilizers, was used in the study either as untreated or anaerobically preserved raw material. Addition of pectinases increased the hydrolysis yield by 26%, 54%, and 64% from the theoretical carbohydrates of untreated, acid, and alkali-preserved materials, respectively. Steam explosion and hot alkali treatment increased the conversion of the total carbohydrates by 78% and 60%, respectively, compared to the untreated hemp. Elevated separation of cells within the hemp stalk tissues and an increased surface area was revealed after hot alkali or pectinase treatments, contributing to the increased conversion to sugars by commercial enzymes.

  10. Response of enzymatic and non-enzymatic antioxidant defense systems of Polygonum hydropiper to Mn stress

    Institute of Scientific and Technical Information of China (English)

    杨贤均; 邓冬梅; 刘可慧; 于方明


    The response of enzyme and non-enzymatic antioxidants of Mn hyperaccumuator,Polygonum hydropiper (P. hydropiper), to Mn stress was studied using hydroponics culture experiments to explore the mechanism of Mn tolerance in this species. Results showed that both chlorophyll and carotenoid contents significantly (p<0.05) decreased with increasing Mn treatment levels (0, 0.5, 1, 2, 4, and 8 mg/L) in hydroponics. The concentrations of malondialdehyde (MDA) and hydrogen peroxide (H2O2) in the root and shoot ofP. hydropiper were accumulated under Mn stress. Meanwhile, the anti-oxidative functions of several important enzymes, including superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and peroxidase (POD) in plants were stimulated by Mn spike in leaves and roots, especially at low Mn stress; while sulfhydryl group (—SH) and glutathion (GSH) were likely involved in Mn detoxification ofP. hydropiper under high Mn stress.


    Directory of Open Access Journals (Sweden)

    A. Vaisi-Raygani


    Full Text Available The etiopathogenesis of dementia in Alzheimer's disease (AD is still unclear. However, long-term oxidative stress is believed to be one of the major contributing factors in progression of neuronal degeneration and decline of cognitive function in AD. In order to assess the presence of oxidative stress in AD, we examined the enzymatic activities of the erythrocyte Cu-Zn superoxide dismutase (Cu-Zn SOD, glutathione peroxidase (GSH-Px, catalase (CAT, and plasma level of total antioxidant status (TAS in AD and control groups (age and sex-matched. The results showed that the Cu-Zn SOD activity was significantly higher and the level of GSH-Px and TAS activities were significantly lower in AD subjects than that in the control group (2111±324 U/grHb, 43.7±11.6 U/grHb, and 1.17 ±0.23 mmol/L compared with 1371±211 U/gHb; t= -2.17, p=0.036, 56.3±9.5 U/gHb; t=3.8, p=0.014, and 1.54±0.2 mmol/L; t=11.18, P<0.001, respectively. While, the erythrocyte CAT activity was lower in AD subjects compared to the control group, the difference was not statistically significant (t=1.3, P=0.15. These findings support the idea that the oxidative stress plays an important role in the pathogenesis underlying AD neurodegeneration. In addition, the enzymatic activity of the erythrocyte Cu-Zn SOD and GSH-Px and the plasma level of TAS can be used as a measure of the oxidative stress and a marker for pathological changes in the brain of patients with AD.

  12. Combined Enzymatic and High-Pressure Processing Affect Cell Wall Polysaccharides in Berries

    NARCIS (Netherlands)

    Hilz, H.; Lille, M.; Poutanen, K.; Schols, H.A.; Voragen, A.G.J.


    The effect of high-pressure processing (HPP) on cell wall polysaccharides in berries was investigated. HPP decreased the degree of methyl esterification (DM), probably by activation of pectin methyl esterase (PME), and improved the extractability of pectins. When commercial enzyme mixtures were adde

  13. Enzymatic pretreatment of lignocellulosic wastes to improve biogas production. (United States)

    Ziemiński, K; Romanowska, I; Kowalska, M


    The effect of enzymatic pretreatment of sugar beet pulp and spent hops prior to methane fermentation was determined in this study. These industrial residues were subjected to enzymatic digestion before anaerobic fermentation because of high fiber content (of 85.1% dry matter (DM) and 57.7% DM in sugar beet pulp and spent hops, respectively). Their 24h hydrolysis with a mix of enzymatic preparations Celustar XL and Agropect pomace (3:1, v/v), with endoglucanase, xylanase and pectinase activities, was most effective. Reducing sugars concentrations in hydrolysates of sugar beet pulp and spent hops were by 88.9% and 59.4% higher compared to undigested materials. The highest yield of biogas was obtained from the enzymatic hydrolysate of sugar beet pulp (183.39 mL/d from 1g COD at fermenter loading with organic matter of 5.43 g COD/L × d). Fermentation of sugar beet pulp gave 19% less biogas. Methane fermentation of spent hops hydrolysate yielded 121.47 mL/d biogas from 1g COD (at 6.02 g COD/L × d, 13% more than from spent hops). These results provide evidence that suitable enzymatic pretreatment of lignocellulosic wastes improve biogas yield from anaerobic fermentation.

  14. Enzymatic activities and effects of mycovirus infection on the virulence of Metarhizium anisopliae in Rhipicephalus microplus. (United States)

    Perinotto, Wendell M S; Golo, Patricia S; Coutinho Rodrigues, Caio J B; Sá, Fillipe A; Santi, Lucélia; Beys da Silva, Walter O; Junges, Angela; Vainstein, Marilene H; Schrank, Augusto; Salles, Cristiane M C; Bittencourt, Vânia R E P


    The present study aimed to evaluate the pathogenic potential of different Metarhizium anisopliae s.l. isolates and to determine whether differences in enzymatic activities of proteases, lipases and chitinases and infection with mycoviruses affect the control of Rhipicephalus microplus achieved by these fungal isolates. Engorged female ticks were exposed to fungal suspensions. The lipolytic and proteolytic activities in the isolates were evaluated using chromogenic substrates and the chitinolytic activity was determined using fluorescent substrates. A gel zymography was performed to determine the approximate size of serine proteases released by M. anisopliae isolates. To detect mycoviral infections, dsRNA was digested using both RNAse A and S1 endonuclease; samples were analyzed on an agarose gel. Four of the five isolates tested were infected with mycovirus; however, the level of control of R. microplus ticks achieved with the only isolate free of infection (isolate CG 347) was low. This finding suggests that mycoviral infection does not affect the virulence of fungi against ticks. Although all five isolates were considered pathogenic to R. microplus, the best tick control and the highest levels of enzymatic activity were achieved with the isolates CG 629 and CG 148. The in vitro activities of lipases, proteases and chitinases produced by M. anisopliae s.l. differed among isolates and may be related to their virulence.

  15. Recent Advances in Enzymatic Fuel Cells: Experiments and Modeling

    Directory of Open Access Journals (Sweden)

    Ivan Ivanov


    Full Text Available Enzymatic fuel cells convert the chemical energy of biofuels into electrical energy. Unlike traditional fuel cell types, which are mainly based on metal catalysts, the enzymatic fuel cells employ enzymes as catalysts. This fuel cell type can be used as an implantable power source for a variety of medical devices used in modern medicine to administer drugs, treat ailments and monitor bodily functions. Some advantages in comparison to conventional fuel cells include a simple fuel cell design and lower cost of the main fuel cell components, however they suffer from severe kinetic limitations mainly due to inefficiency in electron transfer between the enzyme and the electrode surface. In this review article, the major research activities concerned with the enzymatic fuel cells (anode and cathode development, system design, modeling by highlighting the current problems (low cell voltage, low current density, stability will be presented.

  16. Production of prebiotic oligosaccharides by novel enzymatic catalysis

    DEFF Research Database (Denmark)

    Nordvang, Rune Thorbjørn; Jers, Carsten

    evidence backing them up. Since chemical synthesis of carbohydrates is extremely cumbersome, it is generally accepted that HMOs must be produced biochemically and enzymatic in vitro production is a popular strategy. Thus, the purpose of this PhD project was to encompass as many of the aspects...... of the enzymatic production of HMOs as possible, and identify opportunities to improve the enzymes, reaction efficiencies and processes involved. For enzymatic in vitro production of HMOs, industrial side stream products are often used as substrates to reduce the final product price. However, to use...... mono-sialylated HMO, 3’sialyllactose(3’SL), received particular attention in this PhD project. The BioEng state of the art trans-sialidase was, during this PhD project, further mutated, raising the bar for competing enzymes. For further improvement of the current leading enzyme, it was concluded...

  17. Feasibility of pressurization to speed up enzymatic hydrolysis of biological materials for multielement determinations. (United States)

    Moreda-Piñeiro, Antonio; Bermejo-Barrera, Adela; Bermejo-Barrera, Pilar; Moreda-Piñeiro, Jorge; Alonso-Rodríguez, Elia; Muniategui-Lorenzo, Soledad; López-Mahía, Purificación; Prada-Rodríguez, Darío


    The feasibility of pressurized solvents (liquids at a high pressure and/or high temperature without the subcritical point being reached) has been newly investigated to accelerate enzymatic hydrolysis processes of mussel tissue for multielement determinations. The target elements (Al, As, Cd, Co, Cu, Fe, Hg, Li, Mn, Pb, Se, Sr, V, and Zn) were released from dried mussel tissue by action of two proteases (pepsin and pancreatin), and they have been evaluated by inductively coupled plasma optical emission spectrometry (ICP-OES). Variables inherent to the enzymatic activity (pH, ionic strength, temperature, and enzyme mass) and factors affecting pressurization (static time, pressure, and number of cycles) were simultaneously studied by applying a Plackett-Burman design (PBD) as the screening method. Results showed that pH, ionic strength, and temperature were the most statistically significant factors (confidence interval of 95%) under pressurized conditions for pepsin, while pH and ionic strength affected pancreatin activity. This means that metal extraction is mostly attributed to enzymatic activity. The static time (enzymatic hydrolysis time) was found statistically nonsignificant for most of the elements, meaning that the hydrolysis procedure can be finished within a 2-15 min range. For pepsin, optimized conditions (pH 1.0, temperature 40 degrees C, pressure 1500 psi, static time 2 min, and number of cycles 3) gave quantitative extractions for As, Cd, Co, Cu, Hg, Li, Mn, Pb, Se, Sr, V, and Zn. The pepsin mass was 0.05 g, and the solution was Milli-Q water at pH 1.0 (adjusted with hydrochloric acid). For pancreatin, quantitative recoveries were only reached for As, Cd, Cu, Li, Pb, and Sr at room temperature, at a pressure of 1500 psi, for a static time of 2 min and a number of cycles of 3. The extraction solution was a 0.3 M potassium dihydrogen phosphate/potassium hydrogen phosphate buffer at a pH of 7.5 working at room temperature. Around 0.5 g of diatomaceous

  18. Utilization of hydrolysate from lignocellulosic biomass pretreatment to generate electricity by enzymatic fuel cell system. (United States)

    Kim, Sung Bong; Kim, Dong Sup; Yang, Ji Hyun; Lee, Junyoung; Kim, Seung Wook


    The waste hydrolysate after dilute acid pretreatment (DAP) of lignocellulosic biomass was utilized to generate electricity using an enzymatic fuel cell (EFC) system. During DAP, the components of biomass containing hemicellulose and other compounds are hydrolyzed, and glucose is solubilized into the dilute acid solution, called as the hydrolysate liquid. Glucose oxidase (GOD) and laccase (Lac) were assembled on the electrode of the anode and cathode, respectively. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were measured, and the maximum power density was found to be 1.254×10(3) μW/cm(2). The results indicate that the hydrolysate from DAP is a reliable electrolyte containing the fuel of EFC. Moreover, the impurities in the hydrolysate such as phenols and furans slightly affected the charge transfer on the surface of the electrode, but did not affect the power generation of the EFC system in principal.

  19. Cloning and expression of the enzymatic region of Streptococcal hyaluronidase

    Directory of Open Access Journals (Sweden)

    Nafiseh Al-Sadat Mirjamali


    Full Text Available Objective(s: Streptococcus pyogenes produces extracellular hyaluronidase enzyme. This enzyme is directly associated with the spread of the organism during infection. The objective of the present study was to clone and express the nucleotide sequence of the enzyme which is involved in hyaluronidase enzymatic activity. Materials and Methods: The enzymatic region of hyaluronidase gene was detected by bioinformatics method. The PCR method was used to amplify enzymatic region of hyaluronidase gene from chromosomal DNA of Streptococcus pyogenes. The eluted product was cloned into the prokaryotic expression vector pET32a which was digested by BamHI and HindIII restriction endonuclease enzymes. The target protein was expressed in the Escherichia coli. The bacteria including pET32a-hylA (hylA is abbreviation of Streptococcus pyogenes hyaluronidase gene and hylA is abbreviation of Streptococcus pyogenes hyaluronidase protein plasmids were induced by IPTG and analyzed by SDS-PAGE. The enzymatic evaluation and antigenicity was finally studied. Results: Enzymes digestion analysis, sequencing results showed that the target gene (1296 base pair was inserted correctly into the recombinant vector. The expressed protein (65 KDa was purified successfully via affinity chromatography. Data also indicated that enzymatic region of hyaluronidase protein from Streptococcus pyogenes was recognized in all 5 patient’s sera. Conclusion: In general, it is possible to produce the enzymatic regions of the Streptococcus pyogenes hyaluronidase in E. coli. The antigenic property of the produced protein is well retained. Considering the product's domestic demand and also low efficiency of production and pathogenicity of Streptococcus species, it is possible to produce it as recombinant product.

  20. Moving towards a Competitive Fully Enzymatic Biodiesel Process

    Directory of Open Access Journals (Sweden)

    Silvia Cesarini


    Full Text Available Enzymatic biodiesel synthesis can solve several problems posed by the alkaline-catalyzed transesterification but it has the drawback of being too expensive to be considered competitive. Costs can be reduced by lipase improvement, use of unrefined oils, evaluation of soluble/immobilized lipase preparations, and by combination of phospholipases with a soluble lipase for biodiesel production in a single step. As shown here, convenient natural tools have been developed that allow synthesis of high quality FAMEs (EN14214 from unrefined oils in a completely enzymatic single-step process, making it fully competitive.


    Institute of Scientific and Technical Information of China (English)

    Shoujuan Wang; Menghua Qin; Yingjuan Fu; Zhiyong Shao


    The enzymatic deinking and fiber modification of old newsprint (ONP) with several cellulases and xylanase were investigated and the suitable enzyme candidates were selected for ONP deinking in this paper. The results demonstrated that the cellulases and hemicellulases could significantly improve the deinking efficiency and fiber modification.Moreover, the synergistic effects of Novozym342 and xylanase (HC) can further enhance the deinking performance, reduce the dirt count and improve the brightness of resulting pulp. Additionally, compared to deinked pulps, obtained from conventional chemical materials, enzymatically deinked pulps had better bleachability, and the brightness of the bleached pulp reached 59.1% ISO, 9% ISO higher than the unbleached pulp.

  2. Enzymatic network for production of ether amines from alcohols

    DEFF Research Database (Denmark)

    Palacio, Cyntia M.; Crismaru, Ciprian G.; Bartsch, Sebastian


    We constructed an enzymatic network composed of three different enzymes for the synthesis of valuable ether amines. The enzymatic reactions are interconnected to catalyze the oxidation and subsequent transamination of the substrate and to provide cofactor recycling. This allows production...... for reactions containing 10mM alcohol and up to 280mM ammonia corresponded well to predicted conversions. The results indicate that efficient amination can be driven by high concentrations of ammonia and may require improving enzyme robustness for scale-up....

  3. Optimization of Substrate Feeding for Enzymatic Biodiesel Production

    DEFF Research Database (Denmark)

    Price, Jason Anthony; Huusom, Jakob Kjøbsted; Nordblad, Mathias

    to be effective in mitigating the effects of substrate inhibition. Using enzymatic biodiesel production as a case study, the volumetric productivity of the reactor is increased while minimizing inactivation of the enzyme due to the alcohol. This is done by using a simple optimization routine where the substrate...... (both the vegetable oil and alcohol) feed rate/concentration is manipulated simultaneously. The results of the simulation were tested in the laboratory and are sufficiently positive to suggest the implementation of a feeding strategy for large scale enzymatic biodiesel production....

  4. Optimization of Substrate Feeding for Enzymatic Biodiesel Production

    DEFF Research Database (Denmark)

    Price, Jason Anthony; Huusom, Jakob Kjøbsted; Nordblad, Mathias


    to be effective in mitigating the effects of substrate inhibition. Using enzymatic biodiesel production as a case study, the volumetric productivity of the reactor is increased while minimizing inactivation of the enzyme due to the alcohol. This is done by using a simple optimization routine where the substrate...... (both the vegetable oil and alcohol) feed rate/concentration is manipulated simultaneously. The results of the simulation were tested in the laboratory and are sufficiently positive to suggest the implementation of a feeding strategy for large scale enzymatic biodiesel production...

  5. Enzymatic reactors for biodiesel synthesis: Present status and future prospects. (United States)

    Poppe, Jakeline Kathiele; Fernandez-Lafuente, Roberto; Rodrigues, Rafael C; Ayub, Marco Antônio Záchia


    Lipases are being extensively researched for the production of biodiesel as a "silver bullet" in order to avoid the drawbacks of the traditional alkaline transesterification. In this review, we analyzed the main factors involved in the enzymatic synthesis of biodiesel, focusing in the choice of the immobilization protocol, and the parameters involved in the choice and configuration of the reactors. An extensive discussion is presented about the advantages and disadvantages of each type of reactor and their mode of operation. The current scenario of the market for enzymatic biodiesel and some future prospects and necessary developments are also briefly presented.

  6. Escalation of error catastrophe for enzymatic self-replicators (United States)

    Obermayer, B.; Frey, E.


    It is a long-standing question in origin-of-life research whether the information content of replicating molecules can be maintained in the presence of replication errors. Extending standard quasispecies models of non-enzymatic replication, we analyze highly specific enzymatic self-replication mediated through an otherwise neutral recognition region, which leads to frequency-dependent replication rates. We find a significant reduction of the maximally tolerable error rate, because the replication rate of the fittest molecules decreases with the fraction of functional enzymes. Our analysis is extended to hypercyclic couplings as an example for catalytic networks.


    Institute of Scientific and Technical Information of China (English)

    ShoujuanWang; MenghuaQin; YingjuanFu; ZhiyongShao


    The enzymatic deinking and fiber modification of old newsprint (ONP) with several cellulases and xylanase were investigated and the suitable enzyme candidates were selected for ONP deinking in this paper. The results demonstrated that the cellulases and hemicellulases could significantly improve the deinking efficiency and fiber modification. Moreover, the synergistic eflbcts of Novozym342 and xylanase (HC) can further enhance the deinking performance, reduce the dirt count and improve the brightness of resulting pulp. Additionally, compared to deinked pulps, obtained from conventional chemical materials, enzymatically deinked pulps had better bleachability, and the brightness of the bleached pulp reached 59.1% ISO, 9% ISO higher than the unbleached pulp.

  8. Enzymatic Treatment for Shrink-proofing of Wool

    Institute of Scientific and Technical Information of China (English)

    阮建东; 戴瑾瑾


    An alkaline protease was applied on wool knitted fabric in an attempt to improve its shrink resistance. The influence of process parameters was evaluated and the tensile properties, surface characteristics and whiteness of wool were also studied. The results showed that the action of the protease on wool was significantly enhanced by hydrogen peroxide pretreatment. Based on the preliminary test and subsequent factor analysis, an appropriate combined process was obtained. The SEM investigation of the surface characteristics demonstrated that the enzymatic action was not uniform. In addition to the antifelting effects, the enzymatic treatment could lead to an improved degree of whiteness.

  9. Evaluation of wet oxidation pretreatment for enzymatic hydrolysis of softwood

    DEFF Research Database (Denmark)

    Palonen, H.; Thomsen, A.B.; Tenkanen, M.;


    The wet oxidation pretreatment (water, oxygen, elevated temperature, and pressure) of softwood (Picea abies) was investigated for enhancing enzymatic hydrolysis. The pretreatment was preliminarily optimized. Six different combinations of reaction time, temperature, and pH were applied......, and the compositions of solid and liquid fractions were analyzed. The solid fraction after wet oxidation contained 58-64% cellulose, 2-16% hemicellulose, and 24-30% lignin. The pretreatment series gave information about the roles of lignin and hemicellulose in the enzymatic hydrolysis. The temperature...

  10. Aluminium, beta-amyloid and non-enzymatic glycosylation. (United States)

    Exley, C; Schley, L; Murray, S; Hackney, C M; Birchall, J D


    The non-enzymatic glycosylation of beta-amyloid is implicated in the aetiology of Alzheimer's disease. However, controversy surrounds the nature of any involvement and a potential mechanism has not been fully elucidated. We present evidence of an aluminium-induced aggregation of the A beta P(25-35) peptide and speculate that the mechanism of formation of our ordered beta-amyloid aggregates might involve non-enzymatic glycosylation and/or site-specific crosslinking of beta-amyloid fibrils by atomic aluminium.

  11. Effects of Marine Toxins on the Reproduction and Early Stages Development of Aquatic Organisms

    Directory of Open Access Journals (Sweden)

    Vítor Ramos


    Full Text Available Marine organisms, and specially phytoplankton species, are able to produce a diverse array of toxic compounds that are not yet fully understood in terms of their main targets and biological function. Toxins such as saxitoxins, tetrodotoxin, palytoxin, nodularin, okadaic acid, domoic acid, may be produced in large amounts by dinoflagellates, cyanobacteria, bacteria and diatoms and accumulate in vectors that transfer the toxin along food chains. These may affect top predator organisms, including human populations, leading in some cases to death. Nevertheless, these toxins may also affect the reproduction of aquatic organisms that may be in contact with the toxins, either by decreasing the amount or quality of gametes or by affecting embryonic development. Adults of some species may be insensitive to toxins but early stages are more prone to intoxication because they lack effective enzymatic systems to detoxify the toxins and are more exposed to the toxins due to a higher metabolic growth rate. In this paper we review the current knowledge on the effects of some of the most common marine toxins on the reproduction and development of early stages of some organisms.

  12. Affective monitoring: A generic mechanism for affect elicitation

    Directory of Open Access Journals (Sweden)

    Hans ePhaf


    Full Text Available In this paper we sketch a new framework for affect elicitation, which is based on previous evolutionary and connectionist modeling and experimental work from our group. Affective monitoring is considered a local match-mismatch process within a module of the neural network. Negative affect is raised instantly by mismatches, incongruency, disfluency, novelty, incoherence, and dissonance, whereas positive affect follows from matches, congruency, fluency, familiarity, coherence, and resonance, at least when an initial mismatch can be solved quickly. Affective monitoring is considered an evolutionary-early conflict and change detection process operating at the same level as, for instance, attentional selection. It runs in parallel and imparts affective flavour to emotional behavior systems, which involve evolutionary-prepared stimuli and action tendencies related to for instance defensive, exploratory, attachment, or appetitive behavior. Positive affect is represented in the networks by high-frequency oscillations, presumably in the gamma band. Negative affect corresponds to more incoherent lower-frequency oscillations, presumably in the theta band. For affect to become conscious, large-scale synchronization of the oscillations over the network and the construction of emotional experiences are required. These constructions involve perceptions of bodily states and action tendencies, but also appraisals as well as efforts to regulate the emotion. Importantly, affective monitoring accompanies every kind of information processing, but conscious emotions, which result from the later integration of affect in a cognitive context, are much rarer events.

  13. Bacterial and enzymatic bioassays for toxicity testing in the environment. (United States)

    Bitton, G; Koopman, B


    Microbioassays using bacteria or enzymes are increasingly applied to measure chemical toxicity in the environment. Attractive features of these assays may include low cost, rapid response to toxicants, high sample throughput, modest laboratory equipment and space requirements, low sample volume, portability, and reproducible responses. Enzymatic tests rely on measurement of either enzyme activity or enzyme biosynthesis. Dehydrogenases are the enzymes most used in toxicity testing. Assay of dehydrogenase activity is conveniently carried out using oxidoreduction dyes such as tetrazolium salts. Other enzyme activity tests utilize ATPases, esterases, phosphatases, urease, luciferase, beta-galactosidase, protease, amylase, or beta-glucosidase. Recently, the inhibition of enzyme (beta-galactosidase, tryptophanase, alpha-glucosidase) biosynthesis has been explored as a basis for toxicity testing. Enzyme biosynthesis was found to be generally more sensitive to organic chemicals than enzyme activity. Bacterial toxicity tests are based on bioluminescence, motility, growth, viability, ATP, oxygen uptake, nitrification, or heat production. An important aspect of bacterial tests is the permeability of cells to environmental toxicants, particularly organic chemicals of hydrophobic nature. Physical, chemical, and genetic alterations of the outer membrane of E. coli have been found to affect test sensitivity to organic toxicants. Several microbioassays are now commercially available. The names of the assays and their basis are: Microtox (bioluminescence), Polytox (respiration), ECHA Biocide Monitor (dehydrogenase activity), Toxi-Chromotest (enzyme biosynthesis), and MetPAD (enzyme activity). An important feature common to these tests is the provision of standardized cultures of bacteria in freeze-dried form. Two of the more recent applications of microbioassays are in sediment toxicity testing and toxicity reduction evaluation. Sediment pore water may be assayed directly or

  14. Synthesis of poly(aminoamides)via enzymatic means (United States)

    Poly(aminoamides) constitute a subclass of polyamides that are water-soluble and useful for several applications. Commercially they are made via chemical reaction pathways. A review is made in this work of the enzymatic approaches towards their syntheses. Lipases and esterases have been found to ...

  15. The Preparation and Enzymatic Hydrolysis of a Library of Esters (United States)

    Sanford, Elizabeth M.; Smith, Traci L.


    An investigative case study involving the preparation of a library of esters using Fischer esterification and alcoholysis of acid chlorides and their subsequent enzymatic hydrolysis by pig liver esterase and orange peel esterase is described. Students work collaboratively to prepare and characterize the library of esters and complete and evaluate…

  16. Enzymatic Breakdown of Type II Collagen in the Human Vitreous

    NARCIS (Netherlands)

    van Deemter, Marielle; Pas, Hendri H.; Kuijer, Roel; van der Worp, Roelofje J.; Hooymans, Johanna M. M.; Los, Leonoor I.


    PURPOSE. To investigate whether enzymatic collagen breakdown is an active process in the human vitreous. METHODS. Human donor eyes were used for immunohistochemistry to detect the possible presence of the matrix metalloproteinase (MMP)-induced type II collagen breakdown product col2-3/4C-short in th

  17. Coupled chemo(enzymatic reactions in continuous flow

    Directory of Open Access Journals (Sweden)

    Ruslan Yuryev


    Full Text Available This review highlights the state of the art in the field of coupled chemo(enzymatic reactions in continuous flow. Three different approaches to such reaction systems are presented herein and discussed in view of their advantages and disadvantages as well as trends for their future development.

  18. Enzymatic Synthesis of a CCK-8 Tetrapeptide Fragment

    Institute of Scientific and Technical Information of China (English)

    Guang Ya XIANG; Heiner ECKSTEIN


    The enzymatic synthesis of a tetrapeptide Phac-Met-Gly-Trp-Met-OEt is reported. It was synthesized by coupling Phac-Met-OEt with Gly-OMe>·HCl, Trp-OMe and Met-OEt successively, catalyzed by α-chymotrypsin, papain and α-chymotrypsin respectively. The results of FAB-MS showed that the products had the correct molecular mass.

  19. Enhancement of Enzymatic Saccharification of Poplar by Green Liquor Pretreatment

    Directory of Open Access Journals (Sweden)

    Xin Meng


    Full Text Available Green liquor (Na2S + Na2CO3, GL pretreatment is an effective pathway for improving the enzymatic digestibility of lignocellulosic biomass for the production of bioethanol. In this work, GL was employed as a pretreatment to enhance the enzymatic saccharification of poplar. During pretreatment, the increase of H-factor and TTA charge resulted in enhanced delignification and increased degradation of polysaccharides. The sugar yield of enzymatic hydrolysis increased rapidly with increasing TTA charge in GL pretreatment, while the effect of different H-factors (from 400 to 800 on sugar yield was unnoticeable. The pretreated solid recovery was 75.5% at a lignin removal rate of 29.2% under optimized conditions of total titratable alkali (TTA charge 20%, sulfidity 25%, and H-factor 400. The sugar yield of glucan, xylan, and total sugar of GL-pretreated poplar in enzymatic hydrolysis reached up to 89.9%, 65.5%, and 82.8%, respectively, at a cellulase loading of 40 FPU/g-cellulose.

  20. Functional palm oil-based margarine by enzymatic interesterification

    DEFF Research Database (Denmark)

    Ibrahim, Nuzul Amri Bin; Xu, Xuebing

    Palm stearin, palm kernel and fish oils were blended to a various composition ratios and enzymatically interesterified by Lipozyme TL IM lipase (Thermomyces lanuginosa) using a continuous packed bed reactor. The ratio of the oils ranged from 60-90%, 10-40% and 0-10% respectively. The enzyme...

  1. Performance evaluation of six homocysteine cycle enzymatic detection systems

    Institute of Scientific and Technical Information of China (English)



    Objective To evaluate the performance of six homocysteine(Hcy)analysis systems.Methods This is a methodological evaluation study.We analyzed six cycle enzymatic systems,and their correlation and deviation compared with chemiluminescence microparticle immunoassay(CMIA)from Abbott Architect plus i2000 system,Precision,accuracy,anti-interference and analytical

  2. The Mechanisms of Plant Cell Wall Deconstruction during Enzymatic Hydrolysis

    DEFF Research Database (Denmark)

    Thygesen, Lisbeth Garbrecht; E. Thybring, Emil; Johansen, Katja Salomon


    Mechanical agitation during enzymatic hydrolysis of insoluble plant biomass at high dry matter contents is indispensable for the initial liquefaction step in biorefining. It is known that particle size reduction is an important part of liquefaction, but the mechanisms involved are poorly understo...

  3. Drug interactions between inhaled corticosteroids and enzymatic inhibitors


    Daveluy, Amélie; Raignoux, Cécile; Miremont-Salamé, Ghada; Girodet, Pierre-Olivier; Moore, Nicholas; Haramburu, Françoise; Molimard, Mathieu


    Drug interactions between inhaled corticosteroids and enzymatic inhibitors phone: +33-557-571561 (Daveluy, Amelie) (Daveluy, Amelie) Centre Regional de Pharmacovigilance, Hopital Pellegrin - 33076 - Bordeaux Cedex - FRANCE (Daveluy, Amelie) Unite 657, INSERM - Bordeaux - FRANCE (Daveluy, Amelie) Departement de Pharmacologie, CHU de Bordeaux - Bordeaux - FRANCE (Daveluy, Amelie) Centre Regional de Pharmacovigilance, Hopital Pellegrin - 33076...

  4. Nonthermal effect of microwave irradiation in nonaqueous enzymatic esterification. (United States)

    Wan, Hui-da; Sun, Shi-yu; Hu, Xue-yi; Xia, Yong-mei


    Microwave has nonthermal effects on enzymatic reactions, mainly caused by the polarities of the solvents and substrates. In this experiment, a model reaction with caprylic acid and butanol that was catalyzed by lipase from Mucor miehei in alkanes or arenes was employed to investigate the nonthermal effect in nonaqueous enzymatic esterification. With the comparison of the esterification carried by conventional heating and consecutive microwave irradiation, the positive nonthermal effect on the initial reaction rates was found substrate concentration-dependent and could be vanished ostensibly when the substrate concentration was over 2.0 mol L(-1). The polar parameter log P well correlates the solvent polarity with the microwave effect, comparing to dielectric constant and assayed solvatochromic solvent polarity parameters. The log P rule presented in conventional heating-enzymatic esterification still fits in the microwaved enzymatic esterification. Alkanes or arenes with higher log P provided positive nonthermal effect in the range of 2 ≤ log P ≤ 4, but yielded a dramatic decrement after log P = 4. Isomers of same log P with higher dielectric constant received stronger positive nonthermal effect. With lower substrate concentration, the total log P of the reaction mixture has no obvious functional relation with the microwave effect.

  5. Highly Stable Foams from Block Oligomers Synthesized by Enzymatic Reactions

    NARCIS (Netherlands)

    Sagis, L.M.C.; Boeriu, C.G.; Frissen, A.E.; Schols, H.A.; Wierenga, P.A.


    We have synthesized a new amphiphilic block oligomer by the enzymatic linking of a fatty acid (lauric acid) to a fructan oligomer (inulin) and tested the functionality of this carbohydrate derivative in foam stabilization. The structure of the modified oligosaccharide was found to be (Fruc)n(Glc)1CO

  6. Non-enzymatic palladium recovery on microbial and synthetic surfaces

    DEFF Research Database (Denmark)

    Rotaru, Amelia-Elena; Jiang, Wei; Finster, Kai


    in the presence of cells as compared to cell-free controls. We found no difference between native (untreated) and autoclaved cells, and could demonstrate that even a non-enzymatic protein (bovine serum albumin) stimulated Pd(II) reduction as efficiently as bacterial cells. Amine groups readily interact with Pd...

  7. Enzymatic generation of hydrogen peroxide shows promising antifouling effect

    DEFF Research Database (Denmark)

    Kristensen, J.B.; Olsen, Stefan Møller; Laursen, B.S.;


    The antifouling (AF) potential of hydrogen peroxide (H2O2) produced enzymatically in a coating containing starch, glucoamylase, and hexose oxidase was evaluated in a series of laboratory tests and in-sea field trials. Dissolved H2O2 inhibited bacterial biofilm formation by eight of nine marine...


    Directory of Open Access Journals (Sweden)

    Zhiqiang Li,


    Full Text Available Bamboo is a potential lignocellulosic biomass for the production of bioethanol because of its high cellulose and hemicelluloses content. In this research, ethanol organosolv pretreatment with dilute sulfuric acid as the catalyst was studied in order to enhance enzymatic saccharification of moso bamboo. The addition of 2% (w/w bamboo dilute sulfuric acid in 75% ethanol had a particularly strong effect on fractionation of bamboo. It yielded a solids fraction containing 83.4% cellulose in the treated substrate. The cellulose conversion to glucose yield reached 77.1 to 83.4% after enzymatic hydrolysis of the solids fraction for 48 h at an enzyme loading of 15 FPU cellulase/g cellulose and 30 IU β-glucosidase/g cellulose. The enzymatic hydrolysis rate was significantly accelerated as the ethanol organosolv pretreatment time increased, reaching the highest enzymatic glucose yield of 83.4% after 48 h at 50 °C. The concentrations of fermentation inhibitors such as HMF (5-hydroxy-2-methyl furfural and furfural were 0.96 g/L and 4.38 g/L in the spent liquor after the ethanol organosolv pretreatment, which were slightly lower than the concentrations quantified during H2SO4-water treatment. Spent liquor was diluted with water, and more than 87.2% of lignin in raw bamboo was recovered as ethanol organosolv lignin through the filtration process.

  9. Hot biological catalysis: isothermal titration calorimetry to characterize enzymatic reactions. (United States)

    Mazzei, Luca; Ciurli, Stefano; Zambelli, Barbara


    Isothermal titration calorimetry (ITC) is a well-described technique that measures the heat released or absorbed during a chemical reaction, using it as an intrinsic probe to characterize virtually every chemical process. Nowadays, this technique is extensively applied to determine thermodynamic parameters of biomolecular binding equilibria. In addition, ITC has been demonstrated to be able of directly measuring kinetics and thermodynamic parameters (kcat, KM, ΔH) of enzymatic reactions, even though this application is still underexploited. As heat changes spontaneously occur during enzymatic catalysis, ITC does not require any modification or labeling of the system under analysis and can be performed in solution. Moreover, the method needs little amount of material. These properties make ITC an invaluable, powerful and unique tool to study enzyme kinetics in several applications, such as, for example, drug discovery. In this work an experimental ITC-based method to quantify kinetics and thermodynamics of enzymatic reactions is thoroughly described. This method is applied to determine kcat and KM of the enzymatic hydrolysis of urea by Canavalia ensiformis (jack bean) urease. Calculation of intrinsic molar enthalpy (ΔHint) of the reaction is performed. The values thus obtained are consistent with previous data reported in literature, demonstrating the reliability of the methodology.

  10. Enzymatic activity of rodents acclimated to cold and long scotophase (United States)

    Fourie, F. Le R.; Haim, A.


    Rodents representative of a diurnal species ( Rhabdomys pumilio) as well as a nocturnal species ( Praomys natalensis) were acclimated to cold (Ta = 8°C) at a photoperiod of LD 12:12 and a long scotophase (LD 8; 16) at a temperature of 25° C(Ta). Control groups were kept for both species at Ta = 25° C and LD 12:12 and winter acclimated individuals were obtained during July and August to serve as further reference. Blood samples obtained from the tail were analysed for enzymes representative of three major biochemical pathways. The enzymatic activity of LDH (glycolytic pathway), MDH (Krebs cycle) and G6PDH (hexose monophosphate shunt, as an indicator of gonadal activity) were monitored to represent metabolic activity of the respective cycles. Cold acclimated as well as winter acclimatized mice revealed similar enzymatic patterns for both species and significant increases in LDH and MDH were recorded with a concurrent decrease in G6PDH activity. Specimens exposed to long scotophase exhibited similar enzymatic patterns for both species studied, but enzymatic activity was higher than those of cold acclimated individuals. From these results it is concluded that cold as well as long scotophase induce metabolic adaptations through biochemical activity in the experimental animals. The effect of long scotophase is assumed to be an important factor in the induction of winter acclimatization.

  11. An improved competitive inhibition enzymatic immunoassay method for tetrodotoxin quantification


    Stokes Amber N; Williams Becky L; French Susannah S


    Abstract Quantifying tetrodotoxin (TTX) has been a challenge in both ecological and medical research due to the cost, time and training required of most quantification techniques. Here we present a modified Competitive Inhibition Enzymatic Immunoassay for the quantification of TTX, and to aid researchers in the optimization of this technique for widespread use with a high degree of accuracy and repeatability.

  12. Galactomannan enzymatic immunoassay cross-reactivity caused by Prototheca species. (United States)

    Van den Bossche, D; De Bel, A; Hendrickx, M; De Becker, A; Jacobs, R; Naessens, A; Piérard, D


    We report a reactive Aspergillus galactomannan enzymatic immunoassay against the serum of a patient with invasive Prototheca zopfii infection. Analysis of the supernatants of suspensions of P. zopfii and other Prototheca isolates revealed positive results as well. These data suggest cross-reactivity with the serum Aspergillus galactomannan assay in invasive protothecosis.

  13. An improved competitive inhibition enzymatic immunoassay method for tetrodotoxin quantification

    Directory of Open Access Journals (Sweden)

    Stokes Amber N


    Full Text Available Abstract Quantifying tetrodotoxin (TTX has been a challenge in both ecological and medical research due to the cost, time and training required of most quantification techniques. Here we present a modified Competitive Inhibition Enzymatic Immunoassay for the quantification of TTX, and to aid researchers in the optimization of this technique for widespread use with a high degree of accuracy and repeatability.

  14. Galactomannan Enzymatic Immunoassay Cross-Reactivity Caused by Prototheca Species


    Van den Bossche, D.; De Bel, A.; Hendrickx, M.; Becker, A.; Jacobs, R.; Naessens, A; Piérard, D.


    We report a reactive Aspergillus galactomannan enzymatic immunoassay against the serum of a patient with invasive Prototheca zopfii infection. Analysis of the supernatants of suspensions of P. zopfii and other Prototheca isolates revealed positive results as well. These data suggest cross-reactivity with the serum Aspergillus galactomannan assay in invasive protothecosis.

  15. An improved competitive inhibition enzymatic immunoassay method for tetrodotoxin quantification. (United States)

    Stokes, Amber N; Williams, Becky L; French, Susannah S


    Quantifying tetrodotoxin (TTX) has been a challenge in both ecological and medical research due to the cost, time and training required of most quantification techniques. Here we present a modified Competitive Inhibition Enzymatic Immunoassay for the quantification of TTX, and to aid researchers in the optimization of this technique for widespread use with a high degree of accuracy and repeatability.

  16. Lime pretreatment and fermentation of enzymatically hydrolyzed sugarcane bagasse. (United States)

    Rabelo, Sarita C; Maciel Filho, Rubens; Costa, Aline C


    Sugarcane bagasse was subjected to lime (calcium hydroxide) pretreatment and enzymatic hydrolysis for second-generation ethanol production. A central composite factorial design was performed to determine the best combination of pretreatment time, temperature, and lime loading, as well as to evaluate the influence of enzymatic loadings on hydrolysis conversion. The influence of increasing solids loading in the pretreatment and enzymatic hydrolysis stages was also determined. The hydrolysate was fermented using Saccharomyces cerevisiae in batch and continuous mode. In the continuous fermentation, the hydrolysates were concentrated with molasses. Lime pretreatment significantly increased the enzymatic digestibility of sugarcane bagasse without the need for prior particle size reduction. In the optimal pretreatment conditions (90 h, 90 °C, 0.47 glime/g bagasse) and industrially realistic conditions of hydrolysis (12.7 FPU/g of cellulase and 7.3 CBU/g of β-glucosidase), 139.6 kglignin/ton raw bagasse and 126.0 kg hemicellulose in the pretreatment liquor per ton raw bagasse were obtained. The hydrolysate from lime pretreated sugarcane bagasse presented low amounts of inhibitors, leading to ethanol yield of 164.1 kgethanol/ton raw bagasse.

  17. Studies on the non-enzymatic glucosylation of human proteins

    Energy Technology Data Exchange (ETDEWEB)

    Brighton, M.W.


    Aspects of the thiobarbituric acid (TBA) method for quantitating non-enzymatic glucosylation (NEG) in proteins were assessed. Levels of NEG determined by this procedure were compared with values obtained by borohydride reduction and (/sup 14/C) labeling methods. Human albumin was non-enzymatically glucosylated in vitro and extent of glucosylation measured by the TBA, borohydride reduction and (/sup 14/C) labeling procedures. Comparison of in vivo and in vitro NEG was made by the TBA and borohydride reduction techniques. Kinetics of NEG of albumin and whole plasma proteins were assessed and compared. Non-enzymatic glucosylation of each of the major plasma protein fractions was demonstrated both in vivo and in vitro. Relative extends of glucosylation were established. A possible source of error when measuring total plasma NEG in patients with disturbed albumin/globulin ratios is described. Reversibility of the glucose-protein interaction was demonstrated in vitro. Evidence supporting the resistance of albumin to proteolysis, when non-enzymatically glucosylated, is presented.

  18. Wet explosion pretreatment of sugarcane bagasse for enhanced enzymatic hydrolysis

    DEFF Research Database (Denmark)

    Biswas, Rajib; Uellendahl, Hinrich; Ahring, Birgitte Kiær


    Wet explosion pretreatment of sugarcane bagasse was investigated in pilot-scale with the aim of obtaining the highest possible sugar yield after pretreatment. The temperatures used were 155, 170, 185 and 200 C with or without addition of oxygen (0.6 MPa pressure). Enzymatic hydrolysis of washed...

  19. Methods for Improving Enzymatic Trans-glycosylation for Synthesis of Human Milk Oligosaccharide Biomimetics

    DEFF Research Database (Denmark)

    Zeuner, Birgitte; Jers, Carsten; Mikkelsen, Jørn Dalgaard


    Recently, significant progress has been made within enzymatic synthesis of biomimetic, functional glycans, including, for example, human milk oligosaccharides. These compounds are mainly composed of N-acetylglucosamine, fucose, sialic acid, galactose, and glucose, and their controlled enzymatic...

  20. Comparison of dilute mineral and organic acid pretreatment for enzymatic hydrolysis of wheat straw

    NARCIS (Netherlands)

    Kootstra, A.M.J.; Beeftink, H.H.; Scott, E.L.; Sanders, J.P.M.


    The efficiencies of fumaric, maleic, and sulfuric acid in wheat straw pretreatment were compared. As a measure for pretreatment efficiency, enzymatic digestibility of the lignocellulose was determined. Monomeric glucose and xylose concentrations were measured after subsequent enzymatic hydrolysis, a

  1. The effects of diffuse and distinct affect

    NARCIS (Netherlands)

    Stapel, D.A.; Koomen, W; Ruijs, K.I


    In a series of Suboptimal priming studies, it was shown that both affective and nonaffective reactions to a Stimulus may occur without awareness. Moreover. it was demonstrated that affective information is detected earlier than nonaffective information. Therefore, early reactions to an affect-laden

  2. Ultrasound assisted enzymatic depolymerization of aqueous guar gum solution. (United States)

    Prajapat, Amrutlal L; Subhedar, Preeti B; Gogate, Parag R


    The present work investigates the effectiveness of application of low intensity ultrasonic irradiation for the intensification of enzymatic depolymerization of aqueous guar gum solution. The extent of depolymerization of guar gum has been analyzed in terms of intrinsic viscosity reduction. The effect of ultrasonic irradiation on the kinetic and thermodynamic parameters related to the enzyme activity as well as the intrinsic viscosity reduction of guar gum using enzymatic approach has been evaluated. The kinetic rate constant has been found to increase with an increase in the temperature and cellulase loading. It has been observed that application of ultrasound not only enhances the extent of depolymerization but also reduces the time of depolymerization as compared to conventional enzymatic degradation technique. In the presence of cellulase enzyme, the maximum extent of depolymerization of guar gum has been observed at 60 W of ultrasonic rated power and ultrasonic treatment time of 30 min. The effect of ultrasound on the kinetic and thermodynamic parameters as well as the molecular structure of cellulase enzyme was evaluated with the help of the chemical reaction kinetics model and fluorescence spectroscopy. Application of ultrasound resulted in a reduction in the thermodynamic parameters of activation energy (Ea), enthalpy (ΔH), entropy (ΔS) and free energy (ΔG) by 47%, 50%, 65% and 1.97%, respectively. The changes in the chemical structure of guar gum treated using ultrasound assisted enzymatic approach in comparison to the native guar gum were also characterized by FTIR. The results revealed that enzymatic depolymerization of guar gum resulted in a polysaccharide with low degree of polymerization, viscosity and consistency index without any change in the core chemical structure which could make it useful for incorporation in food products.

  3. Visualizing metabolite distribution and enzymatic conversion in plant tissues by desorption electrospray ionization mass spectrometry imaging. (United States)

    Li, Bin; Knudsen, Camilla; Hansen, Natascha Krahl; Jørgensen, Kirsten; Kannangara, Rubini; Bak, Søren; Takos, Adam; Rook, Fred; Hansen, Steen H; Møller, Birger Lindberg; Janfelt, Christian; Bjarnholt, Nanna


    In comparison with the technology platforms developed to localize transcripts and proteins, imaging tools for visualization of metabolite distributions in plant tissues are less well developed and lack versatility. This hampers our understanding of plant metabolism and dynamics. In this study, we demonstrate that desorption electrospray ionization mass spectrometry imaging (DESI-MSI) of tissue imprints on porous Teflon may be used to accurately image the distribution of even labile plant metabolites such as hydroxynitrile glucosides, which normally undergo enzymatic hydrolysis by specific β-glucosidases upon cell disruption. This fast and simple sample preparation resulted in no substantial differences in the distribution and ratios of all hydroxynitrile glucosides between leaves from wild-type Lotus japonicus and a β-glucosidase mutant plant that lacks the ability to hydrolyze certain hydroxynitrile glucosides. In wild-type, the enzymatic conversion of hydroxynitrile glucosides and the concomitant release of glucose were easily visualized when a restricted area of the leaf tissue was damaged prior to sample preparation. The gene encoding the first enzyme in hydroxynitrile glucoside biosynthesis in L. japonicus leaves, CYP79D3, was found to be highly expressed during the early stages of leaf development, and the hydroxynitrile glucoside distribution in mature leaves reflected this early expression pattern. The utility of direct DESI-MSI of plant tissue was demonstrated using cryo-sections of cassava (Manihot esculenta) tubers. The hydroxynitrile glucoside levels were highest in the outer cell layers, as verified by LC-MS analyses. The unexpected discovery of a hydroxynitrile-derived di-glycoside shows the potential of DESI-MSI to discover and guide investigations into new metabolic routes.

  4. Undesirable Enzymatic Browning in Crustaceans: Causative Effects and Its Inhibition by Phenolic Compounds. (United States)

    Nirmal, Nilesh Prakash; Benjakul, Soottawat; Ahmad, Mehraj; Arfat, Yasir Ali; Panichayupakaranant, Pharkphoom


    Undesirable enzymatic browning mediated by polyphenol oxidase (E.C. on the surface of seafood from crustaceans have been a great concern to food processors, causing quality losses of seafood products. Seafoods especially from crustaceans are worldwide consumed due to their delicacy and nutritional value. However, black spot formation (melanosis) is the major problem occurring in crustaceans during postmortem handling and refrigerated storage induce deleterious changes in organoleptic properties and, therefore, decreases commercial value. Polyphenoloxidase (PPO), the copper-containing metalloprotein involved in oxidation of phenol to quinone is the major biochemical reaction of melanosis formation. This enzymatic mechanism causes unappealing blackening in postharvest crustaceans. To alleviate the melanosis formation in crustaceans, use of phenolic compounds from plant extract can serve as antimelanotics and appears to be a good alternative to the conventional sulfites which are associated with health-related disorders. In this review, we focuses on the unique features about the structure, distribution, and properties of PPO as well as mechanism of melanosis formation and provide a comprehensive deeper insight on the factors affecting melanosis formation and its inhibition by various antimelanotics including newly discovered plant phenolic compounds.

  5. Enzymatic transesterification of palm stearin and olein blends to produce zero-trans margarine fat

    Directory of Open Access Journals (Sweden)

    Sellami Mohamed


    Full Text Available Abstract Background Food industries aim to replace trans fat in their products by formulations having equivalent functionality and economic viability. Enzymatic transesterification can be a technological option to produce trans free fats targeting commercial applications. Results Palm stearin and palm olein blends in different ratios were enzymatically transesterified in a solvent free system using a Rhizopus oryzae lipase immobilised onto CaCO3 to produce a suitable fat for margarine formulation. Slip melting points and triacylglycerols profiles were evaluated upon transesterification. Results indicated that all transesterified blends had lower slip melting points than their non transesterified counterparts. Furthermore, the triacylglycerols profile showed a decrease in the concentration of the high melting point triacylglycerols. The rheological analysis showed that margarine prepared with the transesterified blend showed a better spreadability than that of a control margarine prepared with non transesterified fat. Adding powder of dry bark orange to margarine preparation improved its colour and fairly affected its spreadability and rheological behaviour. The margarine prepared with transesterified fat displayed a rheological behaviour that was comparable to that of commercial sample. Conclusions This study is an ecofriendly approach to the utilization of relatively low value bioresources like palm stearin and palm olein for making margarine free of trans fatty acids that are now implicated as risk factor for heart diseases.

  6. Effects of β-cyclodextrins on the enzymatical hydrolysis of chiral dichlorprop methyl ester

    Institute of Scientific and Technical Information of China (English)

    WEN Yue-zhong; ZHOU Shan-shan; FANG Zhao-hua; LIU Wei-ping


    The effect of β-cyclodextrins(β-CDs) on the enzymatical hydrolysis of chiral dichlorprop methyl ester (DCPPM) was studied.Four kinds of β-cyclodextrins(β-cyclodextrin, Partly methylated-CD( PM-β-CD), hydroxypropyl-cyclodextrin(HP-β-CD) and carboxymethylcyclodextrin(CM-β-CD) ) were used. Compared with 100% DCPPM in the absence of β-cyclodextrins, the activity of lipase decreased with the increase of β-cyclodextrin and PM-β-cyclodextrin. However, CM-β-cyclodextrin stimulated the lipase activity. The inhibition effect of β-cyclodextrin and PM-β-cyclodextrin on the hydrolysis of DCPPM is affected by many factors other than degree of the methylation blocking the active site of lipase. UV-Vis and Fourier transform infrared(FTIR) spectroscopy studies of the complexation of aqueous DCPPM with β-CDs provide fresh insight into the molecular structure of the complex and explain the effects of β-CDs on enzymatical hydrolysis of chiral DCPPM. Data showed that inclusion complexes had formed by complexation of the CM-β-CD with DCPPM and the solubility of DCPPM was increased in water, which leaded to the increased lipase activity.

  7. The effect of chemical, physical and enzymatic treatments on the dewatering of tar sands tailings

    Energy Technology Data Exchange (ETDEWEB)

    Theriault, Y.; Masliyah, J.H.; Fedorak, P.M.; Vazquez-Duhalt, R.; Gray, M.R. [University of Alberta, Edmonton, AB (Canada). Dept. of Chemical Engineering


    Fine tailings (with solids {lt} 106{mu}m) from hot-water extraction of the Athabasca tar sands were subjected to thermal, physical, chemical and enzymatic treatments in an attempt to modify the dewatering characteristics of the solid particles. A low-speed centrifuge at circa 1500 relative centrifugal force for 15 h was used to accelerate the sedimentation of the solids, and allowed comparison of the ultimate concentration of solids after the various treatments. Fine tailings were subjected to thermal treatment, sonication and modification of the water chemistry. Although the rate of sedimentation was affected by these treatments, the ultimate volume fraction of solids after centrifuging was about 0.3 (50-55 wt% solids). Extraction of the tailings to remove bitumen, humins and other organic components also failed to change the ultimate solids concentration. Oxidation of the organic material in the tailings by hydrogen peroxide in Fenton`s reagent, and enzymatic oxidation with lignin peroxidase, manganese peroxidase, cytochrome c, horseradish peroxidase and cowpea peroxidase gave no significant effect on the volume fraction of solids in the sediment after centrifugation. The observation that all of these treated samples gave an ultimate volume fraction of solids of about 0.3 indicated that bitumen, adsorbed organic matter and salts had little effect on the ultimate dewatering of tailings. 26 refs., 3 figs., 5 tabs.

  8. Synthesis and enzymatic hydrolysis of esters, constituting simple models of soft drugs. (United States)

    Graffner-Nordberg, M; Sjödin, K; Tunek, A; Hallberg, A


    One way to minimise systemic side effects of drugs is to design molecules, soft drugs, in such a way that they are metabolically inactivated rapidly after having acted on their pharmacological target. Hydrolases (esterases, peptidases, lipases, glycosidases, etc.) are enzymes well suited to use for drug inactivation since they are ubiquitously distributed. Insertion of ester bonds susceptible to enzymatic cleavage may represent one approach to make the action of a drug more restricted to the site of application. The present study describes the chemical synthesis of fourteen model compounds comprising a bicyclic aromatic unit connected by an ester-containing bridge to another aromatic ring. Initial attempts to define a) the tissue selectivity of the hydrolytic metabolism and b) the molecular structural factors affecting the rate of enzymatic ester cleavage are presented. The data show that human and rat liver fractions were more active than human duodenal mucosa and human blood leukocytes at hydrolysing the compounds. The rank order of the compounds was, however, very similar in the different biological systems. Commercially available pig liver carboxyl esterase and cholesterol esterase both reasonably well predict the rank order in the tissue fractions.

  9. Enzymatic Production of Bioxylitol from Sawdust Hydrolysate: Screening of Process Parameters. (United States)

    Rafiqul, I S M; Sakinah, A M M; Zularisam, A W


    Xylose-rich sawdust hydrolysate can be an economic substrate for the enzymatic production of xylitol, a specialty product. It is important to identify the process factors influencing xylitol production. This research aimed to screen the parameters significantly affecting bioxylitol synthesis from wood sawdust by xylose reductase (XR). Enzymatic bioxylitol production was conducted to estimate the effect of different variables reaction time (2-18 h), temperature (20-70 °C), pH (4.0-9.0), NADPH (1.17-5.32 g/L), and enzyme concentration (2-6 %) on the yield of xylitol. Fractional factorial design was followed to identify the key process factors. The screening design identified that time, temperature, and pH are the most significant factors influencing bioxylitol production among the variables with the values of 12 h, 35 °C, and 7.0, respectively. These conditions led to a xylitol yield of 71 % (w/w). This is the first report on the statistical screening of process variables influencing enzyme-based bioxylitol production from lignocellulosic biomass.

  10. Preparation of 6-APA by Enzymatic Bioconversion in an Emulsion Liquid Membrane Reactor

    Institute of Scientific and Technical Information of China (English)

    陆强; 胡鸣; 熊丹柳; 邓修


    Production of 6-aminopenicillanic acid (6-APA) by hydrolysis using penicillin acylase (PA) was studied as a model of an enzymatic emulsion liquid membrane (ELM) process. The loss of PA activity was examined for various membrane compositions (organic solvent, surfactant, carrier). The effects of some experimental variables on the stability of emulsion were investigated. It was found that the choice of organic solvent greatly affected tilestability of the emulsion. Increasing the concentration of the carrier in the membrane phase increases the transfer rate of substrate and products but also has a destabilizing effect on the emulsion. The recovery of 6-APA obtained by a di-carrier system (N263-N1923) was much higher than those when either of the di-carriers was used separately.The whole process was controlled both by the enzymatic reaction rate and by the transfer rate of the substrate and the products, however, the ratio of them could be changed by varying the composition of the system. For an optimum condition, it was obtained that the recovery ratio of 6-APA was over 80% and the conversion of benzyl penicillin (PG) was up to 90% in the external phase after 30 minutes. Meanwhile, the breakage percentage of the emulsion was less than 2%.

  11. Enzymatic polymerization of aniline in the presence of different inorganic substrates

    Energy Technology Data Exchange (ETDEWEB)

    Flores-Loyola, E. [Centro de Investigacion en Quimica Aplicada, Blvd. Enrique Reyna No. 140, CP 25100 Saltillo, Coah (Mexico); Escuela de Ciencias Biologicas, UA de C. Carr. Torreon-Matamoros Km 7.5, Ciudad Universitaria, CP 27400 Torreon, Coah. (Mexico)], E-mail:; Cruz-Silva, R. [Centro de Investigacion en Ingenieria y Ciencias Aplicadas, UAEM. Av. Universidad 1001, Col. Chamilpa, CP 62210, Cuernavaca Mor. (Mexico); Romero-Garcia, J.; Angulo-Sanchez, J.L. [Centro de Investigacion en Quimica Aplicada, Blvd. Enrique Reyna No. 140, CP 25100 Saltillo, Coah (Mexico); Castillon, F.F.; Farias, M.H. [Centro de Ciencias de la Materia Condensada de la UNAM, Apdo. Postal 2681, CP 22800 Ensenada, B.C. (Mexico)


    The effect of different inorganic substrates in the structure of polyaniline synthesized by enzymatic oxidation was studied. The polymer characterization was done by electronic absorption and X-ray photoelectron spectroscopy. The substrates studied were: controlled pore glass, mordenite, zeolite Y, zeolite MCM-41, Wollastonite, silica gel, fuming silica and short glass fibers type E. Polyaniline was synthesized in the presence of the substrates under acidic aqueous conditions, using hydrogen peroxide as oxidizer and HRP or SBP enzymes as catalyst. The composition of the substrates strongly affected the degree of electronic conjugation of the synthesized polyaniline, whereas the pore size and the enzyme type apparently had no effect. The chemical structure of polyaniline enzymatically synthesized was more sensitive to the substrate composition than that chemically synthesized. Apparently substrates containing alkaline ions, such as sodium and calcium, promoted the formation of the branched, non-conductive polyaniline form. The effect of the substrates on the polyaniline structure can be explained considering the local pH effect of the templates surface on the coupling reaction of aniline radicals.

  12. Lipophilic prodrugs of apomorphine I: preparation, characterisation, and in vitro enzymatic hydrolysis in biorelevant media. (United States)

    Borkar, Nrupa; Li, Boyang; Holm, René; Håkansson, Anders E; Müllertz, Anette; Yang, Mingshi; Mu, Huiling


    Apomorphine, a subcutaneously administered drug for Parkinson's disease with short half-life requires frequent administration leading to patient non-compliance. This study aimed at synthesising and purifying lipophilic diesters of apomorphine, and investigating their in vitro degradation in biorelevant media before and after incorporating them into self-emulsifying drug delivery systems (SEDDS) for oral delivery. Two apomorphine diester prodrugs were synthesised: dilauroyl apomorphine (DLA) and dipalmitoyl apomorphine (DPA). The in vitro enzymatic hydrolysis of diesters was performed using biorelevant media with pancreatin to catalyse the diester degradation. The synthesised and purified diesters were found to be free from reactants as impurities confirmed by LC/MS and NMR. DLA and DPA were degraded into corresponding monoesters and free apomorphine within 5 min after adding pancreatin, leaving about 4% and 28% of the intact diester, respectively. The incorporation of the diesters into SEDDS reduced the enzymatic degradation of diesters. In addition, the chain length of diester and the type of oil used in formulations affected diester hydrolysis. The lipophilic apomorphine diesters were substrates of lipases present in pancreatin, and the degree of diester degradation can be controlled by selecting suitable lipid excipients. Therefore, diesters of apomorphine are promising prodrugs for oral delivery aiming at lymphatic transport.

  13. Optimizing harvest of corn stover fractions based on overall sugar yields following ammonia fiber expansion pretreatment and enzymatic hydrolysis

    Directory of Open Access Journals (Sweden)

    Dale Bruce E


    Full Text Available Abstract Background Corn stover composition changes considerably throughout the growing season and also varies between the various fractions of the plant. These differences can impact optimal pretreatment conditions, enzymatic digestibility and maximum achievable sugar yields in the process of converting lignocellulosics to ethanol. The goal of this project was to determine which combination of corn stover fractions provides the most benefit to the biorefinery in terms of sugar yields and to determine the preferential order in which fractions should be harvested. Ammonia fiber expansion (AFEX pretreatment, followed by enzymatic hydrolysis, was performed on early and late harvest corn stover fractions (stem, leaf, husk and cob. Sugar yields were used to optimize scenarios for the selective harvest of corn stover assuming 70% or 30% collection of the total available stover. Results The optimal AFEX conditions for all stover fractions, regardless of harvest period, were: 1.5 (g NH3 g-1 biomass; 60% moisture content (dry-weight basis; dwb, 90°C and 5 min residence time. Enzymatic hydrolysis was conducted using cellulase, β-glucosidase, and xylanase at 31.3, 41.3, and 3.1 mg g-1 glucan, respectively. The optimal harvest order for selectively harvested corn stover (SHCS was husk > leaf > stem > cob. This harvest scenario, combined with optimal AFEX pretreatment conditions, gave a theoretical ethanol yield of 2051 L ha-1 and 912 L ha-1 for 70% and 30% corn stover collection, respectively. Conclusion Changing the proportion of stover fractions collected had a smaller impact on theoretical ethanol yields (29 - 141 L ha-1 compared to the effect of altering pretreatment and enzymatic hydrolysis conditions (150 - 462 L ha-1 or harvesting less stover (852 - 1139 L ha-1. Resources may be more effectively spent on improving sustainable harvesting, thereby increasing potential ethanol yields per hectare harvested, and optimizing biomass processing rather than

  14. Production of Biodiesel by Enzymatic Transesterification: Review

    Directory of Open Access Journals (Sweden)

    Abdel E. Ghaly


    Full Text Available Problem Statement: The research on the production of biodiesel has increased significantly in recent years because of the need for an alternative fuel which endows with biodegradability, low toxicity and renewability. Plant oils, animal fats, microalgal oils and waste products such as animal rendering, fish processing waste and cooking oils have been employed as feedstocks for biodiesel production. In order to design an economically and environmentally sustainable biodiesel production process, a proper understanding of the factors affecting the process and their relative importance is necessary. Approach: A comprehensive review of the literature on the subject of biodiesel production was carried out. Traditionally biodiesel has been produced using either acid or base catalysts. The multi-step purification of end products, wastewater treatment and energy demand of the conventional process has lead to search for alternative option for production of biodiesel. The use the enzyme lipase as a biocatalyst for the transesterification reaction step in biodiesel production has been extensively investigated. Lipase is produced by all living organisms and can be used intracellularly or extracellularly. Conclusion: To date, the most popular microbes used for their lipases have been filamentous fungi and recombinant bacteria. A summary of lipases used in transesterification and their optimum operating conditions is provided. In addition to the choice of lipase employed, factors which make the transesterification process feasible and ready for commercialization are: enzyme modification, the selection of feedstock and alcohol, use of common solvents, pretreatment of the lipase, alcohol to oil molar ratio, water activity/content and reaction temperature. Optimization of these parameters is necessary in order to reduce the cost of biodiesel production. Use of no/low cost waste materials as feedstocks will have double environmental benefits by reducing the

  15. Rheumatoid arthritis affecting temporomandibular joint. (United States)

    Sodhi, Amandeep; Naik, Shobha; Pai, Anuradha; Anuradha, Ardra


    Rheumatoid arthritis (RA) is a chronic, systemic, autoimmune inflammatory disorder that is characterized by joint inflammation, erosive properties and symmetric multiple joint involvement. Temporomandibular joint (TMJ) is very rare to be affected in the early phase of the disease, thus posing diagnostic challenges for the dentist. Conventional radiographs fail to show the early lesions due to its limitations. More recently cone-beam computed tomography (CBCT) has been found to diagnose the early degenerative changes of TMJ and hence aid in the diagnosis of the lesions more accurately. Our case highlights the involvement of TMJ in RA and the role of advanced imaging (CBCT) in diagnosing the bony changes in the early phase of the disease.

  16. Rheumatoid arthritis affecting temporomandibular joint

    Directory of Open Access Journals (Sweden)

    Amandeep Sodhi


    Full Text Available Rheumatoid arthritis (RA is a chronic, systemic, autoimmune inflammatory disorder that is characterized by joint inflammation, erosive properties and symmetric multiple joint involvement. Temporomandibular joint (TMJ is very rare to be affected in the early phase of the disease, thus posing diagnostic challenges for the dentist. Conventional radiographs fail to show the early lesions due to its limitations. More recently cone-beam computed tomography (CBCT has been found to diagnose the early degenerative changes of TMJ and hence aid in the diagnosis of the lesions more accurately. Our case highlights the involvement of TMJ in RA and the role of advanced imaging (CBCT in diagnosing the bony changes in the early phase of the disease.

  17. Synthesis of biodiesel from edible and non-edible oils in supercritical alcohols and enzymatic synthesis in supercritical carbon dioxide

    Energy Technology Data Exchange (ETDEWEB)

    Vivek Rathore; Giridhar Madras [Indian Institute of Science, Bangalore (India). Department of Chemical Engineering


    Biodiesel is an attractive alternative fuel because it is environmentally friendly and can be synthesized from edible and non-edible oils. The synthesis of biodiesel from edible oils like palm oil and groundnut oil and from crude non-edible oils like Pongamia pinnata and Jatropha curcas was investigated in supercritical methanol and ethanol without using any catalyst from 200 to 400{sup o}C at 200 bar. The variables affecting the conversion during transesterification, such as molar ratio of alcohol to oil, temperature and time were investigated in supercritical methanol and ethanol. Biodiesel was also synthesized enzymatically with Novozym-435 lipase in presence of supercritical carbon dioxide. The effect of reaction variables such as temperature, molar ratio, enzyme loading and kinetics of the reaction was investigated for enzymatic synthesis in supercritical carbon dioxide. Very high conversions (>80%) were obtained within 10 min and nearly complete conversions were obtained at within 40 min for the synthesis of biodiesel in supercritical alcohols. However, conversions of only 60-70% were obtained in the enzymatic synthesis even after 8 h. 48 refs., 8 figs., 1 tab.

  18. An antiapoptotic role for telomerase RNA in human immune cells independent of telomere integrity or telomerase enzymatic activity. (United States)

    Gazzaniga, Francesca S; Blackburn, Elizabeth H


    Telomerase is a ribonucleoprotein complex that adds telomeric DNA to the ends of linear chromosomes. It contains two core canonical components: the essential RNA component, hTR, which provides the template for DNA synthesis, and the reverse transcriptase protein component, hTERT. Low telomerase activity in circulating peripheral blood mononuclear cells has been associated with a variety of diseases. It is unknown, however, whether telomerase, in addition to its long-term requirement for telomere maintenance, is also necessary for short-term immune cell proliferation and survival. We report that overexpression of enzymatically inactive hTR mutants protected against dexamethasone-induced apoptosis in stimulated CD4 T cells. Furthermore, hTR knockdown reproducibly induced apoptosis in the absence of any detectable telomere shortening or DNA damage response. In contrast, hTERT knockdown did not induce apoptosis. Strikingly, overexpression of hTERT protein caused apoptosis that was rescued by overexpression of enzymatically inactive hTR mutants. Hence, we propose that hTR can function as a noncoding RNA that protects from apoptosis independent of its function in telomerase enzymatic activity and long-term telomere maintenance in normal human immune cells. These results imply that genetic or environmental factors that alter hTR levels can directly affect immune cell function to influence health and disease.

  19. The effects of centralised and specialised combined pharmacological and psychological intervention compared with decentralised and non-specialised treatment in the early course of severe unipolar and bipolar affective disorders--design of two randomised clinical trials

    DEFF Research Database (Denmark)

    Kessing, Lars Vedel; Hansen, Hanne Vibe; Christensen, Ellen Margrethe


    In unipolar, and bipolar affective disorders, there is a high risk of relapse that increases as the number of episodes increases. Naturalistic follow-up studies suggest that the progressive development of the diseases is not prevented with the present treatment modalities. It is not known whether...

  20. Effects of fibrillation on the wood fibers' enzymatic hydrolysis enhanced by mechanical refining. (United States)

    Liu, Wei; Wang, Bing; Hou, Qingxi; Chen, Wei; Wu, Ming


    The hardwood bleached kraft pulp (HBKP) fibers were pretreated by PFI mill to obtain the substrates, the effects of fibrillation on HBKP fibers' enzymatic hydrolysis was studied. The results showed that the enzymatic hydrolysis efficiency was enhanced obviously by mechanical refining. The mechanical refining alterated the fibers' characteristics such as fibrillation degree, specific surface area, swelling ability, crystallinity, fiber length and fines content. All these factors correlating to the enzymatic hydrolysis were evaluated through mathematical analysis. Among these factors, the fibrillation degree has the profoundest impact on the enzymatic hydrolysis of wood fibers. Consequently, the mechanical refining aiming for a high fibrillation degree was feasible to enhance the enzymatic hydrolysis of lignocellulosic biomass.

  1. Enzymatic ring-opening copolymerization of trimethylene carbonate and ethylene ethyl phosphate

    Institute of Scientific and Technical Information of China (English)

    FENG; Jun; (冯俊); ZHUO; Renxi; (卓仁禧); HE; Feng; (贺枫)


    Enzymatic ring-opening copolymerization of trimethylene carbonate (TMC) and ethylene ethyl phosphate (EEP) are performed in bulk at 100℃ using porcine pancreas lipase (PPL) or candida rugosa lipase (CL) as catalyst. The factors affecting the yield and molecular weights such as catalyst concentration, polymerization time and monomer feed ratio are investigated. The random copolymers obtained have molecular weight ranging from 3200 to 10200. The glass transition temperature (Tg) of the copolymers decreases from 28 to 41.7℃, with the increase of the EEP content in the feed from zero to 5∶10. Degradation tests show that the degradability of the copolymers is improved by introduction of the EEP unit into the copolymer chain.

  2. New enzymatic method for the determination of total phenolic content in tea and wine. (United States)

    Stevanato, Roberto; Fabris, Sabrina; Momo, Federico


    A new spectrophotometric enzymatic method for the determination of total phenol content in tea and wine has been developed. The method is based on the peroxidase-catalyzed oxidation, by hydrogen peroxide, of phenols to phenoxyl radicals, which can react with aromatic substrates to form intensely colored adducts. In comparison with the widely used Folin-Ciocalteu method, this method appears to be more specific and more rapid and as a whole is not affected by the common interfering substances such as ascorbate, citrate, and sulfite. Numerous samples of teas and wines were analyzed by using the new method, and the results compared with those obtained by using the Folin and scavenging of DPPH methods. The differences of the total phenols content found by applying the three methods are discussed in terms of the different specificities of the analytical basis.

  3. Enzymatic cybernetics: an unpublished work by Jacques Monod. (United States)

    Gayon, Jean


    In 1959, Jacques Monod wrote a manuscript entitled Cybernétique enzymatique [Enzymatic cybernetics]. Never published, this unpublished manuscript presents a synthesis of how Monod interpreted enzymatic adaptation just before the publication of the famous papers of the 1960s on the operon. In addition, Monod offers an example of a philosophy of biology immersed in scientific investigation. Monod's philosophical thoughts are classified into two categories, methodological and ontological. On the methodological side, Monod explicitly hints at his preferences regarding the scientific method in general: hypothetical-deductive method, and use of theoretical models. He also makes heuristic proposals regarding molecular biology: the need to analyse the phenomena in question at the level of individual cells, and the dual aspect of all biological explanation, functional and evolutionary. Ontological issues deal with the notions of information and genetic determinism, "cellular memory", the irrelevance of the notion of "living matter", and the usefulness of a cybernetic comprehension of molecular biology.

  4. Enhanced enzymatic cellulose degradation by cellobiohydrolases via product removal

    DEFF Research Database (Denmark)

    Ahmadi Gavlighi, Hassan; Meyer, Anne S.; Mikkelsen, Jørn Dalgaard


    Product inhibition by cellobiose decreases the rate of enzymatic cellulose degradation. The optimal reaction conditions for two Emericella (Aspergillus) nidulans-derived cellobiohydrolases I and II produced in Pichia pastoris were identified as CBHI: 52 °C, pH 4.5–6.5, and CBHII: 46 °C, pH 4.......8. The optimum in a mixture of the two was 50 °C, pH 4.9. An almost fourfold increase in enzymatic hydrolysis yield was achieved with intermittent product removal of cellobiose with membrane filtration (2 kDa cut-off): The conversion of cotton cellulose after 72 h was ~19 % by weight, whereas the conversion...

  5. Novel investigation of enzymatic biodiesel reaction by isothermal calorimetry

    DEFF Research Database (Denmark)

    Søtoft, Lene Fjerbaek; Westh, Peter; Christensen, Knud V.


    Isothermal calorimetry (ITC) was used to investigate solvent-free enzymatic biodiesel production. The transesterification of rapeseed oil with methanol and ethanol was catalyzed by immobilized lipase Novozym 435 at 40 °C. The aim of the study was to determine reaction enthalpy for the enzymatic...... transesterification and to elucidate the mass transfer and energetic processes taking place. Based on the measured enthalpy and composition change in the system, the heat of reaction at 40 °C for the two systems was determined as −9.8 ± 0.9 kJ/mole biodiesel formed from rapeseed oil and methanol, and −9.3 ± 0.7 k...... thermodynamic properties such as reaction enthalpy and reaction rate, the difficulty in actually measuring the true non-mass-transfer-limited reaction kinetics is exposed by the high time resolution of ITC....

  6. Enzymatic hydrolysis of pretreated barley and wheat straw

    DEFF Research Database (Denmark)

    Rosgaard, Lisa


    pretreatment conditions; hot water extraction and acid- or water impregnation followed by steam explosion showed there were slight differences between the effect of pretreatment conditions in relation to the overall yield from enzymatic hydrolysis. The highest glucose concentration was found for barley straw...... subjected to acid impregnation followed by steam explosion; however when the glucose concentration was related to the glucose potential in the substrates, the highest yield was obtained with hot water extracted. Analysis of the supernatants from the pretreatments by mass spectrometry showed that the water...... feeding strategy to increase the substrate loading in the hydrolysis reaction. The substrate for the enzymatic hydrolysis was primarily steam pretreated wheat and barley straw since these substrates were the primary feedstocks for the Babilafuente Bioethanol process. The initial work showed...

  7. Rational design of functional and tunable oscillating enzymatic networks (United States)

    Semenov, Sergey N.; Wong, Albert S. Y.; van der Made, R. Martijn; Postma, Sjoerd G. J.; Groen, Joost; van Roekel, Hendrik W. H.; de Greef, Tom F. A.; Huck, Wilhelm T. S.


    Life is sustained by complex systems operating far from equilibrium and consisting of a multitude of enzymatic reaction networks. The operating principles of biology's regulatory networks are known, but the in vitro assembly of out-of-equilibrium enzymatic reaction networks has proved challenging, limiting the development of synthetic systems showing autonomous behaviour. Here, we present a strategy for the rational design of programmable functional reaction networks that exhibit dynamic behaviour. We demonstrate that a network built around autoactivation and delayed negative feedback of the enzyme trypsin is capable of producing sustained oscillating concentrations of active trypsin for over 65 h. Other functions, such as amplification, analog-to-digital conversion and periodic control over equilibrium systems, are obtained by linking multiple network modules in microfluidic flow reactors. The methodology developed here provides a general framework to construct dissipative, tunable and robust (bio)chemical reaction networks.

  8. From Fed-batch to Continuous Enzymatic Biodiesel Production

    DEFF Research Database (Denmark)


    In this this paper, we use mechanistic modelling to guide the development of acontinuous enzymatic process that is performed as a fed-batch operation. In this workwe use the enzymatic biodiesel process as a case study. A mechanistic model developedin our previous work was used to determine...... measured components (triglycerides, diglycerides, monoglycerides, free fatty acid and fatty acid methyl esters(biodiesel)) much better than using fed-batch data alone given the smaller residuals. We also observe a reduction in the correlation between the parameters.The model was then used to predict that 5...... reactors are required (with a combined residence time of 30 hours) to reach a final biodiesel concentration within 2 % of the95.6 mass % achieved in a fed-batch operation, for 24 hours....

  9. Process simulation and economical evaluation of enzymatic biodiesel production plant. (United States)

    Sotoft, Lene Fjerbaek; Rong, Ben-Guang; Christensen, Knud V; Norddahl, Birgir


    Process simulation and economical evaluation of an enzymatic biodiesel production plant has been carried out. Enzymatic biodiesel production from high quality rapeseed oil and methanol has been investigated for solvent free and cosolvent production processes. Several scenarios have been investigated with different production scales (8 and 200 mio. kg biodiesel/year) and enzyme price. The cosolvent production process is found to be most expensive and is not a viable choice, while the solvent free process is viable for the larger scale production of 200 mio. kg biodiesel/year with the current enzyme price. With the suggested enzyme price of the future, both the small and large scale solvent free production proved viable. The product price was estimated to be 0.73-1.49 euro/kg biodiesel with the current enzyme price and 0.05-0.75 euro/kg with the enzyme price of the future for solvent free process.

  10. Pretreatment of Agave americana stalk for enzymatic saccharification. (United States)

    Yang, Qiang; Pan, Xuejun


    Agave americana is one of commonly grown agave species but currently less valuable because its large flower stalk cannot be used for producing alcoholic beverage. In the present study, the stalk was pretreated with dilute acid (DA), sulfite (SPORL), and sodium hydroxide (NaOH) to preliminarily assess its potential as feedstock for bioethanol production. The changes of cell wall components during the pretreatments, enzymatic digestibility of the pretreated stalks, and the adsorption of cellulases on the substrates were investigated. Results indicated that the pretreatments significantly improved the enzymatic digestibility of the agave stalk. SPORL pretreatment gave higher substrate and sugar yields, while NaOH pretreated stalk had better digestibility under the investigated conditions. The better hydrolysability of NaOH-pretreated stalk was attributed to low lignin and hemicellulose content and high affinity to cellulases.

  11. Parameters optimization for enzymatic assays using experimental design

    Directory of Open Access Journals (Sweden)

    J. F. M. Burkert


    Full Text Available The conditions for maximization enzymatic activity were determined using experimental design and inulinase from Kluyveromyces marxianus ATCC 16045. The effects of substrate concentration (sucrose and inulin, pH and temperature on inulinase activity were verified using four factorial design and surface response analysis. Using sucrose as substrate. It has bean shown that the effects sucrose on enzymatic activity is not statistically significant and the best condition for the highest activity (110 U/mL was achieved with temperature between 60°C and 68°C and pH between 4.5 and 5.0. Using inulin as substrate it was verified that temperature is the only variable statistically significant and the maximum activity was 7.3 U/mL at temperature between 50°C and 51°C.


    Directory of Open Access Journals (Sweden)

    Sofia-Paulina BALAURE


    Full Text Available This study highlights the problem of soil pollution as the result of human activities. Soil pollutans may be either chemicals or biological in nature. microbial enzymatic activities are often proposed as indicators of environmental stress. The soil samples were submitted by chemical, microbiological and enzymatic analyses. Chemical analyses were been made for determinating the heavy metals. Heavy metals from the forest soil were represented by Cu, Zn, Mn, Ni, Pb, Cd and Cr. To evaluate the concentration in heavy metals from the filtrate, we used a acetylene-nitrous oxide flame atomic absorption spectrophotometry. Potential dehydrogenase activity, the only indicator of the possible sources of pollution, excluded the presence of either chemical or biological pollution. The number of bacteria involved in the biogeochemical cycle of nitrogen in the analyzed soil indicated a high efficiency regarding the mineralization of the organic residues of plant and animal origin.

  13. Spectrin’s chimeric E2/E3 enzymatic activity (United States)

    Goodman, Steven R; Petrofes Chapa, Rachel


    In this minireview, we cover the discovery of the human erythrocyte α spectrin E2/E3 ubiquitin conjugating/ligating enzymatic activity and the specific cysteines involved. We then discuss the consequences when this activity is partially inhibited in sickle cell disease and the possibility that the same attenuation is occurring in multiple organ dysfunction syndrome. We finish by discussing the reasons for believing that nonerythroid α spectrin isoforms (I and II) also have this activity and the importance of testing this hypothesis. If correct, this would suggest that the nonerythroid spectrin isoforms play a major role in protein ubiquitination in all cell types. This would open new fields in experimental biology focused on uncovering the impact that this enzymatic activity has upon protein–protein interactions, protein turnover, cellular signaling, and many other functions impacted by spectrin, including DNA repair. PMID:26283706

  14. Loosening Xyloglucan Accelerates the Enzymatic Degradation of Cellulose in Wood

    Institute of Scientific and Technical Information of China (English)

    Rumi Kaida; Tomomi Kaku; Kei'ichi Baba; Masafumi Oyadomari; Takashi Watanabe; Koji Nishida; Toshiji Kanaya; Ziv Shani; Oded Shoseyov; Takahisa Hayashi


    In order to create trees in which cellulose, the most abundant component in biomass, can be enzymatically hydrolyzed highly for the production of bioethanol, we examined the saccharification of xylem from several transgenic poplars, each overexpressing either xyloglucanase, cellulase, xylanase, or galactanase. The level of cellulose degradation achieved by a cellulase preparation was markedly greater in the xylem overexpressing xyloglucanase and much greater in the xylems overexpressing xylanase and cellulase than in the xylem of the wild-type plant. Although a high degree of degradation occurred in all xylems at all loci, the crystalline region of the cellulose microfibrUs was highly degraded in the xylem overexpressing xyloglucanase. Since the complex between microfibrils and xyloglucans could be one region that is particularly resistant to cellulose degradation, loosening xyloglucan could facilitate the enzymatic hydrolysis of cellulose in wood.


    Directory of Open Access Journals (Sweden)

    Jianxin Jiang


    Full Text Available The enzymatic saccharification of pretreated furfural residues with different lignin content was studied to verify the effect of lignin removal in the hydrolysis process. The results showed that the glucose yield was improved by increasing the lignin removal. A maximum glucose yield of 96.8% was obtained when the residue with a lignin removal of 51.4% was hydrolyzed for 108 h at an enzyme loading of 25 FPU/g cellulose. However, further lignin removal did not increase the hydrolysis. The effect of enzyme loading on the enzymatic hydrolysis was also explored in this work. It was concluded that a high glucose yield of 90% was achieved when the enzyme dosage was reduced from 25 to 15 FPU/g cellulose, which was cost-effective for the sugar and ethanol production. The structures of raw material and delignified samples were further characterized by XRD and scanning electron microscopy (SEM.

  16. Utilizing enzymatic digestion procedures in the bioanalytical laboratory. (United States)

    Szapacs, Matthew; Mencken, Thomas; Williams, Jon; Li, Yi


    In recent years, the use of LC-MS technologies in the bioanlytical laboratory for quantitation of peptide/protein biomarkers and biotherapeutics has increased dramatically. The increased interest is due to the improvement in sensitivity of MS instruments over the last 5-10 years, as well as its proven ability to overcome some common issues associated with immunoassay, namely selectivity and reagent availability. However, large proteins (>10 kDa) chromatograph and ionize poorly. To overcome this challenge, LC-MS/MS workflows for proteins larger than 10 kDa utilize enzymatic digestion procedures with subsequent quantitation of one or more of these enzymatically derived peptides to act as a surrogate for the intact protein. Here, recommendations of digestion technique and potential internal standards are summarized.

  17. Improving biogas production from microalgae by enzymatic pretreatment. (United States)

    Passos, Fabiana; Hom-Diaz, Andrea; Blanquez, Paqui; Vicent, Teresa; Ferrer, Ivet


    In this study, enzymatic pretreatment of microalgal biomass was investigated under different conditions and evaluated using biochemical methane potential (BMP) tests. Cellulase, glucohydrolase and an enzyme mix composed of cellulase, glucohydrolase and xylanase were selected based on the microalgae cell wall composition (cellulose, hemicellulose, pectin and glycoprotein). All of them increased organic matter solubilisation, obtaining high values already after 6h of pretreatment with an enzyme dose of 1% for cellulase and the enzyme mix. BMP tests with pretreated microalgae showed a methane yield increase of 8 and 15% for cellulase and the enzyme mix, respectively. Prospective research should evaluate enzymatic pretreatments in continuous anaerobic reactors so as to estimate the energy balance and economic cost of the process.

  18. Bioremediation of uranium contamination with enzymatic uranium reduction (United States)

    Lovley, D.R.; Phillips, E.J.P.


    Enzymatic uranium reduction by Desulfovibrio desulfuricans readily removed uranium from solution in a batch system or when D. desulfuricans was separated from the bulk of the uranium-containing water by a semipermeable membrane. Uranium reduction continued at concentrations as high as 24 mM. Of a variety of potentially inhibiting anions and metals evaluated, only high concentrations of copper inhibited uranium reduction. Freeze-dried cells, stored aerobically, reduced uranium as fast as fresh cells. D. desulfuricans reduced uranium in pH 4 and pH 7.4 mine drainage waters and in uraniumcontaining groundwaters from a contaminated Department of Energy site. Enzymatic uranium reduction has several potential advantages over other bioprocessing techniques for uranium removal, the most important of which are as follows: the ability to precipitate uranium that is in the form of a uranyl carbonate complex; high capacity for uranium removal per cell; the formation of a compact, relatively pure, uranium precipitate.

  19. Teachers in Early Childhood Policy (United States)

    Kilderry, Anna


    This paper examines teacher accountability and authority in early childhood policy. It reports on data from a study that investigated the influences affecting early childhood teacher decision-making at the preschool level in Victoria, Australia. Using a question raised by Ball "Where are the teachers in all this [policy]?" provided a…

  20. 2-Bromopalmitate reduces protein deacylation by inhibition of acyl-protein thioesterase enzymatic activities.

    Directory of Open Access Journals (Sweden)

    Maria P Pedro

    Full Text Available S-acylation, the covalent attachment of palmitate and other fatty acids on cysteine residues, is a reversible post-translational modification that exerts diverse effects on protein functions. S-acylation is catalyzed by protein acyltransferases (PAT, while deacylation requires acyl-protein thioesterases (APT, with numerous inhibitors for these enzymes having already been developed and characterized. Among these inhibitors, the palmitate analog 2-brompalmitate (2-BP is the most commonly used to inhibit palmitoylation in cells. Nevertheless, previous results from our laboratory have suggested that 2-BP could affect protein deacylation. Here, we further investigated in vivo and in vitro the effect of 2-BP on the acylation/deacylation protein machinery, with it being observed that 2-BP, in addition to inhibiting PAT activity in vivo, also perturbed the acylation cycle of GAP-43 at the level of depalmitoylation and consequently affected its kinetics of membrane association. Furthermore, 2-BP was able to inhibit in vitro the enzymatic activities of human APT1 and APT2, the only two thioesterases shown to mediate protein deacylation, through an uncompetitive mechanism of action. In fact, APT1 and APT2 hydrolyzed both the monomeric form as well as the micellar state of the substrate palmitoyl-CoA. On the basis of the obtained results, as APTs can mediate deacylation on membrane bound and unbound substrates, this suggests that the access of APTs to the membrane interface is not a necessary requisite for deacylation. Moreover, as the enzymatic activity of APTs was inhibited by 2-BP treatment, then the kinetics analysis of protein acylation using 2-BP should be carefully interpreted, as this drug also inhibits protein deacylation.

  1. 3-D Enzymatic Nanomaterial Architectures for Energy Harvesting (United States)


    Logan et al., 2006; Luckarift et al., 2014). Since the chemical energy stored in ubiquitous fuels available in the environment is transformed into...such as enzymatic fuel cells, photo- bio energy harvesting devices, nano-biosensors, bio-actuators and other bio-nano-interfacial architectures is the...fossil fuels dwindling, there is an urgent need to find cheap, renewable, and alternate forms of energy using naturally abundant resources such as

  2. Nanosilver: A Catalyst in Enzymatic Hydrolysis of Starch

    Directory of Open Access Journals (Sweden)

    Falkowska Marta


    Full Text Available Silver nanoparticles are widely used, because of their antimicrobial properties. In this paper, the rate of starch digestion in the presence of nanocatalyst was compared with the rate of reaction without nanosilver. The rate of enzymatic degradation of starch was found to be increased in the presence of silver nanoparticles. It is considered that α-amylase was immobilized onto the surface of nanoparticles.

  3. Monitoring of enzymatic reactions using capillary electrophoresis with conductivity detection



    Capillary electrophoresis combined with contactless conductivity detection allows to separate and detect the ionic species, which are neither UV absorbing nor fluorescent. This thesis focuses on the applications of this method on enzymatic reactions in different analytical tasks. First, the non-ionic species ethanol, glucose, ethyl acetate and ethyl butyrate were made accessible for analysis by capillary electrophoresis via charged products or byproducts obtained in enzymati...

  4. Synthesis of CCK-8 Tetrapeptide Fragment by Enzymatic Method

    Institute of Scientific and Technical Information of China (English)

    XIANG Guangya (项光亚); Heiner Eckstein


    The enzymatic synthesis of a tetrapeptide Phac-Met-Gly-Trp-Met-OEt, a fragment of thecholecystokinin C-terminal octapeptide CCK-8, was reported. This fragment was synthesized bycoupling Phac-Met-OEt with Gly-OMe, Trp-OMe and Met-OEt successively. These three stepswere catalyzed by α-chymotrpsin, Papain and α-chymotrpsin respectively. The results of FAB-MSshowed that all the products had the correct molecular mass.

  5. Mercerization and Enzymatic Pretreatment of Cellulose in Dissolving Pulps



    This thesis deals with the preparation of chemically and/or enzymatically modified cellulose. This modification can be either irreversible or reversible. Irreversible modification is used to prepare cellulose derivatives as end products, whereas reversible modification is used to enhance solubility in the preparation of regenerated cellulose. The irreversible modification studied here was the preparation of carboxymethyl cellulose (CMC) using extended mercerization of a spruce dissolving pulp...


    Directory of Open Access Journals (Sweden)

    Simona Dunca


    Full Text Available In the thermoactinomycete strains subjected to examination the values of their recorded enzymatic activities (i.e. α-amy lase, protease, exo-β-1,4 – glucanase, endo -β-1,4 – glucanase and β-glucosidase were lower in the stationary cultures as compared to the stirred ones. The strain Thermomonospora fusca BB255 was found to be highly cellulase- producing and at the same time able to synthesize α-amy lases and proteases.

  7. Biodiesel production by chemical or enzymatic esterification of sunflower oil

    Energy Technology Data Exchange (ETDEWEB)

    Passarinho, Paula C.; Rosa, M. Fernanda; Oliveira, A.C.; Pingarilho, M.S.; Beirao, S.G.; Vieira, Ana Maria Soares


    In this work, two processes of sunflower oil transesterification, with methanol or ethanol, were studied for biodiesel production: chemical (catalyst- NaOH) and enzymatic (catalyst - rhizomucor miehei lipase). The chemical catalysis proved to be more efficient, having been obtained higher conversion yields and a better quality biodiesel, mainly in the case where methanol was used. The transesterification product had, in all cases, to be purified in order to be used as a diesel substitute.

  8. Enzymatic activity in extracts of allergy-causing astigmatid mites. (United States)

    Morgan, Marjorie S; Arlian, Larry G


    Many of the previously characterized allergens of house dust mites are known to be proteases, and this enzymatic activity is thought to contribute to their allergenicity. Other astigmatid mites, including stored-product mites and the ectoparasitic itch mite, Sarcoptes scabiei De Geer, are also known to be allergenic, but little or nothing is known about their enzymatic activities. The purpose of this study was to characterize the enzymatic activities present in extracts of the parasitic itch mite and from eight other species of free-living astigmatid mites. Extracts were prepared from one parasitic mite (S. scabiei), five stored-product mites (Chortoglyphus arcuatus (Troupeau), Lepidoglyphus destructor (Schrank), Blomia tropicalis Bronswijk, Cock, Oshima, Tyrophagus putrescentiae (Schrank), and Acarus siro L.), and three house dust mites [Dermatophagoidesfarinae Hughes, Dermatophagoides pteronyssinus (Troussart), and Euroglyphus maynei (Cooreman) ]. ApiZym strips were used to screen for the presence of 19 individual enzyme activities. Digestion of nine other substrates was evaluated by spectrophotometric or electrophoretic methods. All mite extracts exhibited some form of phosphatase, esterase, aminopeptidase, and glycosidase activity, although their substrate specificities varied considerably. Itch mite extract did not possess detectable serine peptidase activity nor was it able to hydrolyze gelatin or casein, whereas all other mite extracts exhibited these activities. Storage mite extracts possessed enzymes capable of degrading the widest range of substrates, whereas itch mite extract had the most limited proteolytic capacity. Extracts of nine species of allergy-causing astigmatid mites contain wide and diverse repertoires of enzymatic activities. These catalytic activities may be important contributors to the induction and manifestation of inflammatory and immune responses to mites in patients.

  9. A Conductometric Indium Oxide Semiconducting Nanoparticle Enzymatic Biosensor Array



    We report a conductometric nanoparticle biosensor array to address the significant variation of electrical property in nanomaterial biosensors due to the random network nature of nanoparticle thin-film. Indium oxide and silica nanoparticles (SNP) are assembled selectively on the multi-site channel area of the resistors using layer-by-layer self-assembly. To demonstrate enzymatic biosensing capability, glucose oxidase is immobilized on the SNP layer for glucose detection. The packaged sensor c...

  10. Global Warming Potential Of A Waste Refinery Using Enzymatic Treatment

    DEFF Research Database (Denmark)

    Tonini, Davide; Astrup, Thomas


    and fossil resources. This is especially important with respect to the residual waste (i.e. the remains after source-separation and separate collection) which is typically incinerated or landfilled. In this paper the energy and Global Warming performance of a pilot-scale waste refinery for the enzymatic...... plants and utilization of the liquid fraction for biogas production turned out to be the best options with respect to energy and Global Warming performance....


    Directory of Open Access Journals (Sweden)

    Yalçın Çöpür,


    Full Text Available Turkey annually produces 26 million tons of vegetables and is the third-biggest vegetable producer. After harvest, the waste of vegetable stalks lacking of economic value is burnt or left in the fields, causing environmental pollution. The aim of this study was to examine bioethanol production of a mixture of tomato, pepper, and eggplant stalks using an alternative chemical, sodium borohydrate (NaBH4 in a chemical pretreatment step. Both steam-exploded (SE and dry-milled (DM stalks were chemically pretreated and enzymatically hydrolyzed in this study. Results showed that SE stalks had better enzymatic digestibility compared to DM. NaOH treatment removed the highest amount of lignin (17.1%; SE, 2%, 90 min but also glucose (21.5%; SE, 2%, 90 min from the structure. On the other hand, NaBH4 removed the highest lignin in proportion to glucose for both SE and DM samples. Enzymatically hydrolyzed stalks gave the highest sugar yields of 30.1% (o.d.-dry matter for the SE sample when it was pretreated with 2% NaOH for 30 min.

  12. Microstructural study of pre-treated and enzymatic hydrolyzed bamboo

    Directory of Open Access Journals (Sweden)

    Funsho O. KOLAWOLE


    Full Text Available Bamboo was used as biomass feedstock which was pre-treated using dilute acid hydrolysis followed by enzymatic hydrolysis. The bamboo was mechanical ground to particle sizes 212–500µm, followed by pre-treatment with dilute sulfuric acid at a concentration of 0.5 and 1.0 (%v/v at temperatures of 25, 110, 120, 150 and 200°C with time intervals of 2 and 4 hours. Pre-hydrolyzate was later analyzed for reducing sugar using UV-Vis spectrophotometry. Under the above conditions, a maximum glucose yield of 153.1 mg/g was obtained at 200°C and acid concentrations of 1% for 4 hours. Water insoluble solids obtained were subsequently hydrolyzed with Celluclast (Trichoderma reesi and β-glucosidase (Novozyme 188 for 72 hours. Optical Microscope and ESEM images of bamboo samples were obtained at various stages of pre-treatment and enzymatic hydrolysis. Result reveals a breakdown in the ligno-cellulosic structure of the bamboo during exposure to dilute acid and enzymatic hydrolysis.

  13. Path Sampling Methods for Enzymatic Quantum Particle Transfer Reactions. (United States)

    Dzierlenga, M W; Varga, M J; Schwartz, S D


    The mechanisms of enzymatic reactions are studied via a host of computational techniques. While previous methods have been used successfully, many fail to incorporate the full dynamical properties of enzymatic systems. This can lead to misleading results in cases where enzyme motion plays a significant role in the reaction coordinate, which is especially relevant in particle transfer reactions where nuclear tunneling may occur. In this chapter, we outline previous methods, as well as discuss newly developed dynamical methods to interrogate mechanisms of enzymatic particle transfer reactions. These new methods allow for the calculation of free energy barriers and kinetic isotope effects (KIEs) with the incorporation of quantum effects through centroid molecular dynamics (CMD) and the full complement of enzyme dynamics through transition path sampling (TPS). Recent work, summarized in this chapter, applied the method for calculation of free energy barriers to reaction in lactate dehydrogenase (LDH) and yeast alcohol dehydrogenase (YADH). We found that tunneling plays an insignificant role in YADH but plays a more significant role in LDH, though not dominant over classical transfer. Additionally, we summarize the application of a TPS algorithm for the calculation of reaction rates in tandem with CMD to calculate the primary H/D KIE of YADH from first principles. We found that the computationally obtained KIE is within the margin of error of experimentally determined KIEs and corresponds to the KIE of particle transfer in the enzyme. These methods provide new ways to investigate enzyme mechanism with the inclusion of protein and quantum dynamics.

  14. Cometabolic Enzymatic Transformation of Organic Micropollutants under Methanogenic Conditions. (United States)

    Gonzalez-Gil, Lorena; Carballa, Marta; Lema, Juan M


    Anaerobic digestion (AD) has been shown to have the biological potential to decrease concentrations of several organic micropollutants (OMPs) in sewage sludge. However, the mechanisms and factors behind these biotransformations, which are essential for elucidating the possible transformation products and to foster the complete removal of OMPs via operational strategies, remain unclear. Therefore, this study investigated the transformation mechanisms of 20 OMPs during the methanogenic step of AD with a focus on the role of acetate kinase (AK), which is a key enzyme in methane production. The results from lab-scale methanogenic reactors showed that this step accounts for much of the reported OMP biotransformation in AD. Furthermore, enzymatic assays confirmed that AK transforms galaxolide, naproxen, nonylphenol, octylphenol, ibuprofen, diclofenac, bisphenol A, and triclosan. Except for galaxolide, for which further studies are required to refine conclusions, the OMP's chemical structure was a determinant for AK action because only compounds that contain a carboxyl or hydroxyl group and have moderate steric hindrance were enzymatically transformed, likely by phosphorylation. For these seven compounds, this enzymatic mechanism accounts for 10-90% of the measured methanogenic biotransformation, suggesting that other active enzymes of the AD process are also involved in OMP biotransformation.

  15. Process Evaluation Tools for Enzymatic Cascades Welcome Message

    DEFF Research Database (Denmark)

    Abu, Rohana

    Biocatalysis is attracting significant attention from both academic and industrial scientists due to the excellent capability of enzyme to catalyse selective reactions. Recently, much interest has been shown in the application of enzymatic cascades as a useful tool in organic synthesis to synthes......Biocatalysis is attracting significant attention from both academic and industrial scientists due to the excellent capability of enzyme to catalyse selective reactions. Recently, much interest has been shown in the application of enzymatic cascades as a useful tool in organic synthesis...... improvement and implementation. Hence, the goal of this thesis is to evaluate the process concepts in enzymatic cascades in a systematic manner, using tools such as thermodynamic and kinetic analysis. Three relevant case studies have been used to exemplify the approach. In the first case study, thermodynamic...... the equilibrium positions in the main syntheses. In principle, this strategy could successfully achieve high conversion, using ammonia as the sole reagent used in excess to drive the conversion. The findings herein indicate that quantitatively the possibilities for improving the conversion of thermodynamically...

  16. Enzymatic hydrolysis of protein:mechanism and kinetic model

    Institute of Scientific and Technical Information of China (English)

    Qi Wei; He Zhimin


    The bioreaction mechanism and kinetic behavior of protein enzymatic hydrolysis for preparing active peptides were investigated to model and characterize the enzymatic hydrolysis curves.Taking into account single-substrate hydrolysis,enzyme inactivation and substrate or product inhibition,the reaction mechanism could be deduced from a series of experimental results carried out in a stirred tank reactor at different substrate concentrations,enzyme concentrations and temperatures based on M-M equation.An exponential equation dh/dt = aexp(-bh) was also established,where parameters a and b have different expressions according to different reaction mechanisms,and different values for different reaction systems.For BSA-trypsin model system,the regressive results agree with the experimental data,i.e.the average relative error was only 4.73%,and the reaction constants were determined as Km = 0.0748 g/L,Ks = 7.961 g/L,kd = 9.358/min,k2 =38.439/min,Ea= 64.826 kJ/mol,Ed= 80.031 kJ/mol in accordance with the proposed kinetic mode.The whole set of exponential kinetic equations can be used to model the bioreaction process of protein enzymatic hydrolysis,to calculate the thermodynamic and kinetic constants,and to optimize the operating parameters for bioreactor design.


    Directory of Open Access Journals (Sweden)

    Luís M. R. Solano


    Full Text Available The use of biocatalysts in synthetic chemistry is a conventional methodology for preparing enantiomerically enriched compounds. Despite this fact, the number of experiments in chemical teaching laboratories that demonstrate the potential of enzymes in synthetic organic chemistry is limited. We describe a laboratory experiment in which students synthesized a chiral secondary alcohol that can be used in the preparation of antidepressant drugs. This experiment was conducted by individual students as part of a Drug Synthesis course held at the Pharmacy Faculty, Lisbon University. This laboratory experiment requires six laboratory periods, each lasting four hours. During the first four laboratory periods, students synthesized and characterized a racemic ester using nuclear magnetic resonance spectroscopy and gas chromatography. During the last two laboratory periods, they performed enzymatic hydrolysis resolution of the racemic ester using Candida antarctica lipase B to yield enantiomerically enriched secondary alcohol. Students successfully prepared the racemic ester with a 70%-81% overall yield in three steps. The enzymatic hydrolysis afforded (R- secondary alcohol with good enantioselectivity (90%-95% and reasonable yields (10%-19%. In these experiments, students were exposed to theoretical and practical concepts of aromatic acylation, ketone reduction, esterification, and enzymatic hydrolysis.

  18. Mental Health May Affect Chances Against Cancer (United States)

    ... page: Mental Health May Affect Chances Against Cancer Early research suggests ... Our findings contribute to the evidence that poor mental health might have some predictive capacity for certain physical ...

  19. Early clerkships

    NARCIS (Netherlands)

    Kamalski, Digna M. A.; Ter Braak, Edith W. M. T.; Ten Cate, Olle Th. J.; Borleffs, Jan C. C.


    Background: Early clinical experience is being introduced in innovative, vertically integrated undergraduate medical curricula. While in many cases, this early clinical experience is limited to the presence of patients during lectures, in Utrecht students gain 'hands on' experience of daily clinical

  20. Changes in Enzymatic Activity and Quality Attributes of Late-mature Peaches in Response to Controlled Atmosphere Conditions

    Institute of Scientific and Technical Information of China (English)

    TIAN Shi-ping; XU Yong; JIANG Ai-li; WANG Yi


    Late-mature peaches (Prunus persica L. Batsch, cv. Dongxuemi ) were stored in air,modification atmosphere (MA) and controlled atmospheres (CA) at 1℃ to investigate enzymatic activities,flesh browning, quality attributes and fruit storability during storage periods. Peaches stored in CA conditions showed significant lower activities of polyphenol oxidase (PPO) and peroxidase (POD) compared to that kept in MA and in air during the early period of storage. CA treatments indicated a better result in maintaining fruit firmness, color and titratable acidity than MA and control treatments. But there was no significant difference in vitamin C and total soluble solids of peaches stored in CA, MA and air. Increasing PPO activity obviously stimulated flesh browning. Peach fruits stored in CA conditions showed a lower activity of POD and higher firmness in the early period of storage. The peaches stored in 5%CO2 + 5%O2 and 10%CO2 + 5%O2 for 92 days had a good quality without any off-flavor. The results indicated CA conditions obviously inhibited the enzymatic browning, delayed senescence and maintained quality of late-mature peaches.

  1. Oxidoreductases in early gestational monkey placenta during maternal malarial infection : histochemical localisation

    Directory of Open Access Journals (Sweden)

    Nishi Saxena , P.S.R. Murthy


    Full Text Available Background & objectives: Early gestational malaria is more deleterious than late gestational infection.Still the pathophysiology of maternofoetal organ—the placenta in malaria remains almost unexploredduring early gestation. Present study dealing with oxidoreductases in early gestational placenta duringmaternal malarial infection of Plasmodium cynomolgi bastianellii in rhesus monkeys was anticipatedto provide a better insight into the functional impairment of this organ leading to foetal abnormalities.Methods: Three control and four experimental monkeys (Macaca mulatta were quarantined for onemonth prior to experimentation. Experimental monkeys at 2–2½ months of gestation were inoculatedwith P. cynomolgi bastianellii. On attaining first peak of parasitaemia the placentae were collectedfrom anesthetised animals. The snap-frozen, cryostat sections were subjected to histochemicallocalisation for 3 (or 17 β-hydroxysteroid dehydrogenase (β-HSD [3 (or 17 β-hydroxysteroid:NAD (P+ oxidoreductase, EC hydroxysteroid dehydrogenases] and NADPH-tetrazoliumreductase [NADPH : (acceptor oxidoreductase, EC NADPH-TR]. Comparative microscopyof control and malaria infected placental sections was performed and analysed.Results: A localised decrease in both the enzymes was observed in syncytiotrophoblast layer ofmalaria infected monkey placenta. The areas showing morphological damage of syncytiotrophoblastwere also depicting gross reduction in NADPH-TR activity.Interpretation & conclusion: The altered enzymatic activities [3 (or 17 β-HSD and NADPH-TR] inmalaria infected early gestational monkey placenta have been discussed in the light of placentalfunction. It could be concluded by present studies that these alterations would affect the cellularmetabolism especially steroidogenesis and detoxification process which in turn would affect thenormal development of the foetus as well as maintenance of gestation.

  2. Early Intervention with Labor Affects Postural Related Auxiliary Facilities for Childbirth Outcomes%产程早期体位干预配合相关辅助设施对分娩结局的影响

    Institute of Scientific and Technical Information of China (English)



    目的:探讨产程早期体位干预配合相关辅助设施对分娩结局的影响,为合理指导分娩及预防不良妊娠结局提供依据。方法选择2013年9月~2014年9月我院体检的妊娠期孕妇150例作为观察组。选择2012年6月~2013年6月我院150例均未使用早期体位干预配合相关辅助设施的分娩孕妇作为对照组。对照组采用传统的卧位、半卧位待产;观察组给予分娩球联合体位干预待产。比较两组产程时间、疼痛程度、分娩方式及产后2h阴道流血量。结果观察组顺产率明显高于对照组,疼痛评分级别低于对照组,第一、第二产程时间明显短于对照组,产后2h阴道流血量少于对照组,有显著性差异(P<0.05,P<0.01)。两组第三产程时间比较无显著性差异(P>0.05)。结论产程早期体位干预,配合分娩辅助设施,能缩短第一产程时间。减轻分娩疼痛,提高自然分娩率。%Objective To investigate the impact of the production process of early intervention with postural related aux‐iliary facilities on birth outcomes ,and provide reference for reasonable guide childbirth ,and provide the basis for the preven‐tion of adverse pregnancy outcomes .Methods 150 cases of pregnant women in our hospital from September 2013 to Sep‐tember 2014 were accepted physical examination as the observation group while pregnant women from June 2012 to June 2013 were not using birth with early intervention postural related auxiliary facilities ,as a control group .Traditional supine and semi-recumbent position for pregnant women in control group .Give birth ball position to intervene to be produced in observation group .Compare the labor time ,pain ,delivery mode in two groups .Results By different mode of delivery ,the birth rate in observation group was significantly higher than in control group ,the difference was statistically significant . Childbirth pain scores in observation

  3. Early literacy

    DEFF Research Database (Denmark)

    Jensen, Anders Skriver


    This paper discusses findings from the Danish contribution to the EASE project, a European research project running from 2008 to 2010 on early literacy in relation to the transition from childcare to school. It explores a holistic, inclusive approach to early literacy that resists a narrow...... and schools. The paper also draws on Gee’s (2001, 2003, 2004, 2008) sociocultural approach to literacy, and Honneth’s (2003, 2006) concept of recognition. Emphasizing participation and recognition as key elements, it claims that stakeholders in early liter- acy must pay attention to how diverse early literacy...... opportunities empower children, especially when these opportunities are employed in a project-based learning environ- ment in which each child is able to contribute to the shared literacy events....

  4. Mutations in MDH2, Encoding a Krebs Cycle Enzyme, Cause Early-Onset Severe Encephalopathy. (United States)

    Ait-El-Mkadem, Samira; Dayem-Quere, Manal; Gusic, Mirjana; Chaussenot, Annabelle; Bannwarth, Sylvie; François, Bérengère; Genin, Emmanuelle C; Fragaki, Konstantina; Volker-Touw, Catharina L M; Vasnier, Christelle; Serre, Valérie; van Gassen, Koen L I; Lespinasse, Françoise; Richter, Susan; Eisenhofer, Graeme; Rouzier, Cécile; Mochel, Fanny; De Saint-Martin, Anne; Abi Warde, Marie-Thérèse; de Sain-van der Velde, Monique G M; Jans, Judith J M; Amiel, Jeanne; Avsec, Ziga; Mertes, Christian; Haack, Tobias B; Strom, Tim; Meitinger, Thomas; Bonnen, Penelope E; Taylor, Robert W; Gagneur, Julien; van Hasselt, Peter M; Rötig, Agnès; Delahodde, Agnès; Prokisch, Holger; Fuchs, Sabine A; Paquis-Flucklinger, Véronique


    MDH2 encodes mitochondrial malate dehydrogenase (MDH), which is essential for the conversion of malate to oxaloacetate as part of the proper functioning of the Krebs cycle. We report bi-allelic pathogenic mutations in MDH2 in three unrelated subjects presenting with early-onset generalized hypotonia, psychomotor delay, refractory epilepsy, and elevated lactate in the blood and cerebrospinal fluid. Functional studies in fibroblasts from affected subjects showed both an apparently complete loss of MDH2 levels and MDH2 enzymatic activity close to null. Metabolomics analyses demonstrated a significant concomitant accumulation of the MDH substrate, malate, and fumarate, its immediate precursor in the Krebs cycle, in affected subjects' fibroblasts. Lentiviral complementation with wild-type MDH2 cDNA restored MDH2 levels and mitochondrial MDH activity. Additionally, introduction of the three missense mutations from the affected subjects into Saccharomyces cerevisiae provided functional evidence to support their pathogenicity. Disruption of the Krebs cycle is a hallmark of cancer, and MDH2 has been recently identified as a novel pheochromocytoma and paraganglioma susceptibility gene. We show that loss-of-function mutations in MDH2 are also associated with severe neurological clinical presentations in children.

  5. Pressure Modulation of the Enzymatic Activity of Phospholipase A2, A Putative Membrane-Associated Pressure Sensor. (United States)

    Suladze, Saba; Cinar, Suleyman; Sperlich, Benjamin; Winter, Roland


    Phospholipases A2 (PLA2) catalyze the hydrolysis reaction of sn-2 fatty acids of membrane phospholipids and are also involved in receptor signaling and transcriptional pathways. Here, we used pressure modulation of the PLA2 activity and of the membrane's physical-chemical properties to reveal new mechanistic information about the membrane association and subsequent enzymatic reaction of PLA2. Although the effect of high hydrostatic pressure (HHP) on aqueous soluble and integral membrane proteins has been investigated to some extent, its effect on enzymatic reactions operating at the water/lipid interface has not been explored, yet. This study focuses on the effect of HHP on the structure, membrane binding and enzymatic activity of membrane-associated bee venom PLA2, covering a pressure range up to 2 kbar. To this end, high-pressure Fourier-transform infrared and high-pressure stopped-flow fluorescence spectroscopies were applied. The results show that PLA2 binding to model biomembranes is not significantly affected by pressure and occurs in at least two kinetically distinct steps. Followed by fast initial membrane association, structural reorganization of α-helical segments of PLA2 takes place at the lipid water interface. FRET-based activity measurements reveal that pressure has a marked inhibitory effect on the lipid hydrolysis rate, which decreases by 75% upon compression up to 2 kbar. Lipid hydrolysis under extreme environmental conditions, such as those encountered in the deep sea where pressures up to the kbar-level are encountered, is hence markedly affected by HHP, rendering PLA2, next to being a primary osmosensor, a good candidate for a sensitive pressure sensor in vivo.

  6. Early detection of psychosis

    DEFF Research Database (Denmark)

    Larsen, T. K.; Melle, I.; Auestad, B.


    Background During the last decades we have seen a new focus on early treatment of psychosis. Several reviews have shown that duration of untreated psychosis (DUP) is correlated to better outcome. However, it is still unknown whether early treatment will lead to a better long-term outcome....... This study reports the effects of reducing DUP on 5-year course and outcome.Method During 1997â€"2000 a total of 281 consecutive patients aged >17 years with first episode non-affective psychosis were recruited, of which 192 participated in the 5-year follow-up. A comprehensive early detection (ED) programme...... with public information campaigns and low-threshold psychosis detection teams was established in one healthcare area (ED-area), but not in a comparable area (no-ED area). Both areas ran equivalent treatment programmes during the first 2 years and need-adapted treatment thereafter.Results At the start...

  7. Understanding of alkaline pretreatment parameters for corn stover enzymatic saccharification

    Directory of Open Access Journals (Sweden)

    Chen Ye


    Full Text Available Abstract Background Previous research on alkaline pretreatment has mainly focused on optimization of the process parameters to improve substrate digestibility. To achieve satisfactory sugar yield, extremely high chemical loading and enzyme dosages were typically used. Relatively little attention has been paid to reduction of chemical consumption and process waste management, which has proven to be an indispensable component of the bio-refineries. To indicate alkali strength, both alkali concentration in pretreatment solution (g alkali/g pretreatment liquor or g alkali/L pretreatment liquor and alkali loading based on biomass solids (g alkali/g dry biomass have been widely used. The dual approaches make it difficult to compare the chemical consumption in different process scenarios while evaluating the cost effectiveness of this pretreatment technology. The current work addresses these issues through pretreatment of corn stover at various combinations of pretreatment conditions. Enzymatic hydrolysis with different enzyme blends was subsequently performed to identify the effects of pretreatment parameters on substrate digestibility as well as process operational and capital costs. Results The results showed that sodium hydroxide loading is the most dominant variable for enzymatic digestibility. To reach 70% glucan conversion while avoiding extensive degradation of hemicellulose, approximately 0.08 g NaOH/g corn stover was required. It was also concluded that alkali loading based on total solids (g NaOH/g dry biomass governs the pretreatment efficiency. Supplementing cellulase with accessory enzymes such as α-arabinofuranosidase and β-xylosidase significantly improved the conversion of the hemicellulose by 6–17%. Conclusions The current work presents the impact of alkaline pretreatment parameters on the enzymatic hydrolysis of corn stover as well as the process operational and capital investment costs. The high chemical consumption for alkaline

  8. Multi-parametric MRI characterization of enzymatically degraded articular cartilage. (United States)

    Nissi, Mikko J; Salo, Elli-Noora; Tiitu, Virpi; Liimatainen, Timo; Michaeli, Shalom; Mangia, Silvia; Ellermann, Jutta; Nieminen, Miika T


    Several laboratory and rotating frame quantitative MRI parameters were evaluated and compared for detection of changes in articular cartilage following selective enzymatic digestion. Bovine osteochondral specimens were subjected to 44 h incubation in control medium or in collagenase or chondroitinase ABC to induce superficial collagen or proteoglycan (glycosaminoglycan) alterations. The samples were scanned at 9.4 T for T1 , T1 Gd (dGEMRIC), T2 , adiabatic T1 ρ , adiabatic T2 ρ , continuous-wave T1 ρ , TRAFF2 , and T1 sat relaxation times and for magnetization transfer ratio (MTR). For reference, glycosaminoglycan content, collagen fibril orientation and biomechanical properties were determined. Changes primarily in the superficial cartilage were noted after enzymatic degradation. Most of the studied parameters were sensitive to the destruction of collagen network, whereas glycosaminoglycan depletion was detected only by native T1 and T1 Gd relaxation time constants throughout the tissue and by MTR superficially. T1 , adiabatic T1 ρ , adiabatic T2 ρ , continuous-wave T1 ρ , and T1 sat correlated significantly with the biomechanical properties while T1 Gd correlated with glycosaminoglycan staining. The findings indicated that most of the studied MRI parameters were sensitive to both glycosaminoglycan content and collagen network integrity, with changes due to enzymatic treatment detected primarily in the superficial tissue. Strong correlation of T1 , adiabatic T1ρ , adiabatic T2 ρ , continuous-wave T1 ρ , and T1 sat with the altered biomechanical properties, reflects that these parameters were sensitive to critical functional properties of cartilage. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:1111-1120, 2016.

  9. Pregnancy Exercise Increase Enzymatic Antioxidant In Pregnant Women

    Directory of Open Access Journals (Sweden)

    Wagey Freddy Wagey


    Full Text Available Objectives: Pregnancy is a vulnerable condition to all kinds of "stress", resulting in changes of physiological and metabolic functions. This research aims to determine effect of exercise during pregnancy in increasing enzymatic antioxidant marked by increase of superoxide dismutase (SOD, gluthation peroxidase (GSHPx, and catalase (CAT levels. Methods: Randomized pre and posttest control group design was employed in this study. A number of 66 pregnant women were recruited in this study and grouped into two groups, i.e 30 of them as control group and the rest as treatment group. Pregnancy exercise was performed to all 36 pregnant women from 20 weeks gestation on treatment group. The exercise was performed in the morning for about 30 minutes, twice a weeks. On the other hand, daily activities was sugested for control group. Student’s t-test was then applied to determine the mean different of treatment and control group with 5 % of significant value. Results: This study reveals that there were significantly higher increase of (superoxide dismutase (SOD, gluthation peroxidase (GSHPx, and catalse (CAT levels of treatment group compare to control group. These enzymatic antioxidant increase among these two group were around 1.36 mg/gHb for SOD; 1.14 IU/gHb for GSHPx; and 0.97 IU/gHb for CAT, (p < 0.05.  Clinical outcomes, such as strengten of pelvic muscle and quality of life of treatment group were significantly better compared to control group (p < 0.05. Conclusions: This means that exercise during pregnancy ages of 20 weeks increase enzymatic antioxidant levels SOD, GSHPx, and CAT higher compare to control group without exercise.  

  10. Pregnancy Exercise Increase Enzymatic Antioxidant In Pregnant Women

    Directory of Open Access Journals (Sweden)

    Wagey Freddy Wagey


    Full Text Available Objectives: Pregnancy is a vulnerable condition to all kinds of "stress", resulting in changes of physiological and metabolic functions. This research aims to determine effect of exercise during pregnancy in increasing enzymatic antioxidant marked by increase of superoxide dismutase (SOD, gluthation peroxidase (GSHPx, and catalase (CAT levels. Methods: Randomized pre and posttest control group design was employed in this study. A number of 66 pregnant women were recruited in this study and grouped into two groups, i.e 30 of them as control group and the rest as treatment group. Pregnancy exercise was performed to all 36 pregnant women from 20 weeks gestation on treatment group. The exercise was performed in the morning for about 30 minutes, twice a weeks. On the other hand, daily activities was sugested for control group. Student’s t-test was then applied to determine the mean different of treatment and control group with 5 % of significant value. Results: This study reveals that there were significantly higher increase of (superoxide dismutase (SOD, gluthation peroxidase (GSHPx, and catalse (CAT levels of treatment group compare to control group. These enzymatic antioxidant increase among these two group were around 1.36 mg/gHb for SOD; 1.14 IU/gHb for GSHPx; and 0.97 IU/gHb for CAT, (p < 0.05. Clinical outcomes, such as strengten of pelvic muscle and quality of life of treatment group were significantly better compared to control group (p < 0.05. Conclusions: This means that exercise during pregnancy ages of 20 weeks increase enzymatic antioxidant levels SOD, GSHPx, and CAT higher compare to control group without exercise.

  11. Biological and enzymatic treatment of bisphenol A and other endocrine disrupting compounds: a review. (United States)

    Husain, Qayyum; Qayyum, Shariq


    Bisphenol A is predominantly used as an intermediate in the production of polycarbonate plastics and epoxy resins. Traces of bisphenol A released into the environment can reach into the wastewater and soil via application of sewage sludge from wastewater treatment systems that receive water containing bisphenol A, or from leachate from uncontrolled landfills. In this study we have made an effort to review the work on the presence of bisphenol A and other related endocrine disrupting compounds in the environment and their impact on the life of living organisms including human beings. Bisphenol A has several implications on the health of human beings as well it can also affect the growth of plants and animals. Number of physicochemical methods such as adsorption, membrane based filtration, ozonation, fenton, electrochemical and photochemical degradation has been used for the removal of bisphenol A. However, these methods have some inherent limitations and therefore cannot be used for large scale treatment of such pollutants. The alternative procedures have attracted the attention of environmental scientists. Biological methods are looking quite promising and these procedures are helpful in the complete degradation of bisphenol A and related compounds. Several bacterial, fungal, and algal strains and mixed cultures have successfully been employed for the degradation of bisphenol A. Recently, enzymatic methods have attracted the attention of the environmentalists for the treatment of bisphenol A and other endocrine disrupting compounds. Numerous types of oxidoreductases; laccases, tyrosinases, manganese peroxidase, lignin peroxidase, polyphenol oxidases, horseradish peroxidase and bitter gourd peroxidase have exhibited their potential for the remediation of such types of compounds. The cytochrome P 450 monooxygenases and hemoglobin have also participated in the degradation of bisphenol A and other related endocrine disrupting compounds. Various redox mediators

  12. Enzymatic synthesis of hydrophobic compounds integrated with membrane separation

    Directory of Open Access Journals (Sweden)

    Noworyta Andrzej


    Full Text Available The enzymatic synthesis of a highly hydrophobic product (dipeptide precursor in which the reaction is accompanied by the mass transfer of the reaction product to the organic phase and the substrates to the water phase is considered. Equations describing both continuous and batch processes are formulated. The range of variability in the operating parameters of such a bioreactor is specified, and the correlations reported in the literature to describe mass transfer in the membrane contactor are validated. The proposed process was verified experimentally, and good agreement between the determined and calculated concentrations was obtained in both phases.

  13. Sugar ester surfactants: enzymatic synthesis and applications in food industry. (United States)

    Neta, Nair S; Teixeira, José A; Rodrigues, Lígia R


    Sugar esters are non-ionic surfactants that can be synthesized in a single enzymatic reaction step using lipases. The stability and efficiency of lipases under unusual conditions and using non-conventional media can be significantly improved through immobilization and protein engineering. Also, the development of de novo enzymes has seen a significant increase lately under the scope of the new field of synthetic biology. Depending on the esterification degree and the nature of fatty acid and/or sugar, a range of sugar esters can be synthesized. Due to their surface activity and emulsifying capacity, sugar esters are promising for applications in food industry.

  14. Chemistry enters nucleic acids biology: enzymatic mechanisms of RNA modification. (United States)

    Boschi-Muller, S; Motorin, Y


    Modified nucleotides are universally conserved in all living kingdoms and are present in almost all types of cellular RNAs, including tRNA, rRNA, sn(sno)RNA, and mRNA and in recently discovered regulatory RNAs. Altogether, over 110 chemically distinct RNA modifications have been characterized and localized in RNA by various analytical methods. However, this impressive list of known modified nucleotides is certainly incomplete, mainly due to difficulties in identification and characterization of these particular residues in low abundance cellular RNAs. In DNA, modified residues are formed by both enzymatic reactions (like DNA methylations, for example) and by spontaneous chemical reactions resulting from oxidative damage. In contrast, all modified residues characterized in cellular RNA molecules are formed by specific action of dedicated RNA-modification enzymes, which recognize their RNA substrate with high specificity. These RNA-modification enzymes display a great diversity in terms of the chemical reaction and use various low molecular weight cofactors (or co-substrates) in enzymatic catalysis. Depending on the nature of the target base and of the co-substrate, precise chemical mechanisms are used for appropriate activation of the base and the co-substrate in the enzyme active site. In this review, we give an extended summary of the enzymatic mechanisms involved in formation of different methylated nucleotides in RNA, as well as pseudouridine residues, which are almost universally conserved in all living organisms. Other interesting mechanisms include thiolation of uridine residues by ThiI and the reaction of guanine exchange catalyzed by TGT. The latter implies the reversible cleavage of the N-glycosidic bond in order to replace the initially encoded guanine by an aza-guanosine base. Despite the extensive studies of RNA modification and RNA-modification machinery during the last 20 years, our knowledge on the exact chemical steps involved in catalysis of RNA

  15. Stereochemistry of enzymatic water addition to C=C bonds. (United States)

    Chen, Bi-Shuang; Otten, Linda G; Hanefeld, Ulf


    Water addition to carbon-carbon double bonds using hydratases is attracting great interest in biochemistry. Most of the known hydratases are involved in primary metabolism and to a lesser extent in secondary metabolism. New hydratases have recently been added to the toolbox, both from natural sources or artificial metalloenzymes. In order to comprehensively understand how the hydratases are able to catalyse the water addition to carbon-carbon double bonds, this review will highlight the mechanistic and stereochemical studies of the enzymatic water addition to carbon-carbon double bonds, focusing on the syn/anti-addition and stereochemistry of the reaction.

  16. Organic synthesis - applications in enzymatic studies, catalysis and surface modification

    DEFF Research Database (Denmark)

    Viart, Helene Marie-France

    as liposomes and the enzymatic activity was studied. Hydrolysis (or absence of hydrolysis) was monitored by MALDI-TOF-MS. The results observedin these experiments are compared to MD predictions and confirm them. The second chapter deals with surface functionalization of liposomes. The copper mediated [3...... and a functionality covalently linked to a coupling partner (such as small molecule, peptide, etc.), we investigated the efficiency of the reaction depending on the position of the functional groups (whether on the liposome or on the coupling partner). Our results indicate that the reaction is most efficient when...

  17. Enzymatic Synthesis of a CCK-8 Tripeptide Derivative

    Institute of Scientific and Technical Information of China (English)


    The enzymatic synthesis of CCK-8 tripeptide derivative Phac-Met-Asp(OMe)-Phe-NH2 is reported. Starting with Phac-Met-OCam, we have successfully synthesized the target tripeptide with three free or immobilized enzymes, α-chymotrypsin, papain and thermolysin in reasonable yields. The key steps in this synthesis were the coupling of Phac-Met-OCam and H-Asp(OMe)2 to form Met-Asp peptide bond catalyzed by α-chymotrypsin and the selective hydrolysis of α-ester of Phac-Met-Asp(OMe)2 catalyzed by papain.

  18. Modelling and operation of reactors for enzymatic biodiesel production

    DEFF Research Database (Denmark)

    Price, Jason Anthony

    to the production of high fructose corn syrup, upgrading of fats and oils and biodiesel production to name a few. Despite these examples of industrial enzymatic applications, it is still not “clear cut” how to implement biocatalyst in industry and how best to optimize the processes. This is because the processing...... aspects of the enzyme with reaction/reactor engineering is performed. This strategy is applied to a case study of biodiesel production catalysed by a liquid enzyme formulation. The use of enzymes for biodiesel production is still in its infancy with non-optimized process designs. Furthermore is it unclear...

  19. Bio-based alkyds by direct enzymatic bulk polymerization

    DEFF Research Database (Denmark)

    Nguyen, Hiep Dinh

    Alkyd coating systems have been largely used to preserve exterior wood applications as well as to provide them with a decorative appearance. In the current stage of sustainability concerns, there has been a stronger focus on development and production of bio-based coating components, heading toward....... Moreover, the studies on more sensitive monomers such as itaconic acid in enzymatic polymerization has showed that the method is useful in the production of alkyds from such building blocks, which could not be prepared by the corresponding classical boiling method at high temperature. Such systems...... that their functionalized surface markedly altered their solubility, but provided only moderate improvement in the mechanical properties of the alkyd....

  20. Peptide hydrogelation triggered by enzymatic induced pH switch (United States)

    Cheng, Wei; Li, Ying


    It remains challenging to develop methods that can precisely control the self-assembling kinetics and thermodynamics of peptide hydrogelators to achieve hydrogels with optimal properties. Here we report the hydrogelation of peptide hydrogelators by an enzymatically induced pH switch, which involves the combination of glucose oxidase and catalase with D-glucose as the substrate, in which both the gelation kinetics and thermodynamics can be controlled by the concentrations of D-glucose. This novel hydrogelation method could result in hydrogels with higher mechanical stability and lower hydrogelation concentrations. We further illustrate the application of this hydrogelation method to differentiate different D-glucose levels.

  1. Dynamic modeling and validation of a lignocellulosic enzymatic hydrolysis process

    DEFF Research Database (Denmark)

    Prunescu, Remus Mihail; Sin, Gürkan


    on a demonstration scale reactor. The following novel features are included: the application of the Convection–Diffusion–Reaction equation to a hydrolysis reactor to assess transport and mixing effects; the extension of a competitive kinetic model with enzymatic pH dependency and hemicellulose hydrolysis......; a comprehensive pH model; and viscosity estimations during the course of reaction. The model is evaluated against real data extracted from a demonstration scale biorefinery throughout several days of operation. All measurements are within predictions uncertainty and, therefore, the model constitutes a valuable...... tool to support process optimization, performance monitoring, diagnosis and process control at full-scale studies....

  2. Detecting platform for phenolic compounds-characteristic of enzymatic electrode (United States)

    Cabaj, Joanna; Chyla, Antoni; Jędrychowska, Agnieszka; Olech, Kamila; Sołoducho, Jadwiga


    We report here on simple and universal method for the highly efficient, electrolytic immobilization of tyrosinase (from Agaricus bisporus), for amperometric biosensing purposes. Tyrosinase has been successfully deposited on the surface of thin, ordered films of copolymerized derivative of thiophene (3-methylthiophene/3-thiopheneacetic acid/bis(ethylenedioxythiophene)diphenylamine). The tyrosinase retains well its activity well within the fabricated copolymer matrix. Reduction peaks, observed in cyclic voltammetry at 0.125 to +0.07 V, were attributed to the reduction of enzymatically liberated quinone species. Considering the fact, that immobilization strategy showed high efficiency, obtained results suggest that the method for phenoloxidase immobilization has a great potential for fabrication of bioelectronics' devices.

  3. Fed-Batch Feeding Strategies for Enzymatic Biodiesel Production

    DEFF Research Database (Denmark)

    Price, Jason Anthony; Nordblad, Mathias; Woodley, John


    while improving the biodiesel yield. The best experimental results gave a yield of 703 .76 g FAME L-1 and a reactor productivity of 28.12 g FAME L-1 h-1. In comparison, to reach the same yield, the optimised two step feeding strategy took 6.25 hours less, which equates to an increase the reactor...... of the differences in the interfacial and bulk concentrations of the enzyme. The model is then used to evaluate various feeding strategies to improve the enzymatic biodiesel production. The feeding strategies investigated, gave insight into how the methanol should be fed to potentially mitigate enzyme deactivation...

  4. Engineering aspects of enzymatic fiber solubilization from potato pulp

    DEFF Research Database (Denmark)

    Ravn, Helle Christine

    ), and content of the glycoalkaloids α-solanine and α-chaconine. The monosaccharide composition, DM, and content of α-solanine remained rather unchanged, whereas DAc and content of α-chaconine decreased significantly during the campaign. The seven samples were subjected to the same enzymatic treatment...... suggested that potato pulp produced late in the campaign would be preferable to upgrade, due to the higher yield and lower risk of toxicity from glycoalkaloids. The outcome may be the result of an inherent effect of the higher ripeness of the potatoes late in the campaign. Producing fibers from potato pulp...

  5. Post-deposition early-phase migration and retention behavior of radiocesium in a litter-mineral soil system in a Japanese deciduous forest affected by the Fukushima nuclear accident. (United States)

    Koarashi, Jun; Nishimura, Syusaku; Nakanishi, Takahiro; Atarashi-Andoh, Mariko; Takeuchi, Erina; Muto, Kotomi


    The fate of radiocesium ((137)Cs) derived from the Fukushima nuclear accident and associated radiation risks are largely dependent on its migration and retention behavior in the litter-soil system of Japanese forest ecosystems. However, this behavior has not been well quantified. We established field lysimeters in a Japanese deciduous broad-leaved forest soon after the Fukushima nuclear accident to continuously monitor the downward transfer of (137)Cs at three depths: the litter-mineral soil boundary and depths of 5 cm and 10 cm in the mineral soil. Observations were conducted at two sites within the forest from May 2011 to May 2015. Results revealed similar temporal and depth-wise variations in (137)Cs downward fluxes for both sites. The (137)Cs downward fluxes generally decreased year by year at all depths, indicating that (137)Cs was rapidly leached from the forest-floor litter layer and was then immobilized in the upper (0-5 cm) mineral soil layer through its interaction with clay minerals. The (137)Cs fluxes also showed seasonal variation, which was in accordance with variations in the throughfall and soil temperature at the sites. There was no detectable (137)Cs flux at a depth of 10 cm in the mineral soil in the third and fourth years after the accident. The decreased inventory of mobile (or bioavailable) (137)Cs observed during early stages after deposition indicates that the litter-soil system in the Japanese deciduous forest provides only a temporary source for (137)Cs recycling in plants.

  6. Optimization of Process Parameters and Kinetic Model of Enzymatic Extraction of Polyphenols from Lonicerae Flos


    Kong,Fansheng; Yu, Shujuan; Bi, Yongguang; Huang, Xiaojun; Huang, Mengqian


    Objective: To optimize and verify the cellulase extraction of polyphenols from honeysuckle and provide a reference for enzymatic extracting polyphenols from honeysuckle. Materials and Methods: The uniform design was used According to Fick's first law and kinetic model, fitting analysis of the dynamic process of enzymatic extracting polyphenols was conducted. Results: The optimum enzymatic extraction parameters for polyphenols from honeysuckle are found to be 80% (v/v) of alcohol, 35:1 (mL/g) ...

  7. Early Smoking, Education, and Labor Market Performance

    NARCIS (Netherlands)

    Palali, Ali


    This study investigates the effects of early smoking on educational attainment and labor market performance. The results show that early smoking adversely affects educational attainment and initial labor market performance, but only for males. The effect of early smoking on initial labor market perf

  8. Early infection and prognosis after acute stroke

    DEFF Research Database (Denmark)

    Kammersgaard, L P; Jørgensen, H S; Reith, J;


    Infection is a frequent complication in the early course of acute stroke and may adversely affect stroke outcome. In the present study, we investigate early infection developing in patients within 3 days of admission to the hospital and its independent relation to recovery and stroke outcome....... In addition, we identify predictors for early infections, infection subtypes, and their relation to initial stroke severity....

  9. Non-Enzymatic Decomposition of Collagen Fibers by a Biglycan Antibody and a Plausible Mechanism for Rheumatoid Arthritis

    Energy Technology Data Exchange (ETDEWEB)

    Antipova, Olga; Orgel, Joseph P.R.O. (IIT)


    Rheumatoid arthritis (RA) is a systemic autoimmune inflammatory and destructive joint disorder that affects tens of millions of people worldwide. Normal healthy joints maintain a balance between the synthesis of extracellular matrix (ECM) molecules and the proteolytic degradation of damaged ones. In the case of RA, this balance is shifted toward matrix destruction due to increased production of cleavage enzymes and the presence of (autoimmune) immunoglobulins resulting from an inflammation induced immune response. Herein we demonstrate that a polyclonal antibody against the proteoglycan biglycan (BG) causes tissue destruction that may be analogous to that of RA affected tissues. The effect of the antibody is more potent than harsh chemical and/or enzymatic treatments designed to mimic arthritis-like fibril de-polymerization. In RA cases, the immune response to inflammation causes synovial fibroblasts, monocytes and macrophages to produce cytokines and secrete matrix remodeling enzymes, whereas B cells are stimulated to produce immunoglobulins. The specific antigen that causes the RA immune response has not yet been identified, although possible candidates have been proposed, including collagen types I and II, and proteoglycans (PG's) such as biglycan. We speculate that the initiation of RA associated tissue destruction in vivo may involve a similar non-enzymatic decomposition of collagen fibrils via the immunoglobulins themselves that we observe here ex vivo.

  10. Mechanistic insight into sono-enzymatic degradation of organic pollutants with kinetic and thermodynamic analysis. (United States)

    Malani, Ritesh S; Khanna, Swati; Chakma, Sankar; Moholkar, Vijayanand S


    In this paper, we have attempted to get a physical insight into process of sono-enzymatic treatment for degradation of recalcitrant organic pollutants. Decolourization of an azo dye has been used as model reaction with different experimental protocols that alter characteristics of ultrasound and cavitation phenomena in the system. Experimental data is analyzed to determine kinetic and thermodynamic parameters of decolorization process. The trends observed in kinetic and thermodynamic parameters of decolourization are essentially manifestations of the dominating mechanism of the decolorization of the textile dye (or nature of prevalent chemical reaction in the system), viz. either molecular reaction due to enzyme or radical reaction due to transient cavitation. The activation energy for sonochemical protocol is negative, which indicates instantaneity of the radical reactions. The frequency factor is also low, which is attributed to high instability of radicals. For enzymatic and sono-enzymatic protocols, activation energy is positive with higher frequency factor. Enthalpy change for sonochemical protocol is negative, while that for enzymatic and sono-enzymatic protocols is positive. The net entropy change for sonochemical protocol is more negative than enzymatic or sono-enzymatic protocol due to differences in prevalent chemical mechanism of dye decolorization. Due to inverse variations of frequency factor and activation energy, marginal rise in reaction kinetics is seen for sono-enzymatic protocol, as compared to enzymatic treatment alone. Due to inverse variations of enthalpy and entropy change, net Gibbs energy change in all experimental protocols shows little variation indicating synergism of the mechanism of ultrasound and enzyme.

  11. Enzymatic and non-enzymatic antioxidant potentials of Chlorella vulgaris grown in effluent of a confectionery industry. (United States)

    Kumar, R Ranjith; Rao, P Hanumantha; Subramanian, V V; Sivasubramanian, V


    Enzymatic and non-enzymatic antioxidant potentials of Chlorella vulgaris have gained considerable importance in recent decades. C. vulgaris strain highly tolerant to extreme pH variations was isolated and mass-cultivated in the wastewater from a confectionery industry. C.vulgaris showed better growth in wastewater than in improvised CFTRI medium. The microalgal biomass was then screened for the following antioxidants: peroxidase, superoxide dismutase, polyphenol oxidase, glutathione peroxidase, chlorophyll a, ascorbic acid, α-tocopherol and reduced glutathione. The total polyphenol content of the strain was also studied. The strain showed a high degree of enzymatic antioxidant activity (0.195 × 10(-5) ± 0.0072 units/cell peroxidase, 0.04125 × 10(-5) ± 0.001 units/cell superoxide dismutase, 0.2625 × 10(-5) ± 0.003 units/cell polyphenol oxidase and 0.025 × 10(-5) ± 0.003 glutathione peroxidase). The microalgal biomass also showed, per milligram weight, 0.2182 ± 0.005 μg of ascorbic acid, 0.00264 ± 0.001 μg of α-tocopherol and 0.07916 ± 0.004 μg of reduced glutathione. These results represent the possibility of using C. vulgaris grown in confectionery industry wastewater as a source of nutritious supplement, which is highly promising in terms of both economic and nutritional point of view.

  12. Enzymatic detection of formalin-fixed museum specimens for DNA analysis and enzymatic maceration of formalin-fixed specimens

    DEFF Research Database (Denmark)

    Sørensen, Margrethe; Redsted Rasmussen, Arne; Simonsen, Kim Pilkjær


    % ethanol. The method was subsequently tested on wild-living preserved specimens and an archived specimen. The protease enzyme used was SavinaseH 16 L, Type EX from Novozymes A/S. The enzymatic screening test demands only simple laboratory equipment. The method is useful for natural history collections...... in museums where DNA analyses of archived specimens are performed. Wasted time and resources can be avoided through the detection of formalin-fixed specimens because these specimens yield low-quality, damaged DNA. In addition to the screening method, it is shown that formalin-preserved specimens can...

  13. Enzymatic activity of the cellulolytic complex produced by Trichoderma reesei. Enzymatic hydrolysis of cellulose; Actividad enzimatica del complejo celulolitico producido por Trichoderma reesei. Hidrolisis enzimatica de la celulosa

    Energy Technology Data Exchange (ETDEWEB)

    Alfonsel, M.; Negro, M. J.; Saez, R.; Martin, C.


    The enzymatic activity characterization of the cellulolytic complex obtained from Trichoderma reesei QM 9414 and the influence of the enzymatic hydrolysis conditions on the hydrolysis yield are studied. Pure cellulose and native or alkali pretreated biomass Onopordum nervosum have been used as substrates. The values of pH, temperature, substrate concentration and enzyme-substrate ratio for the optimum activity of that complex, evaluated as glucose and reducing sugars production, have been selected. Previous studies on enzymatic hydrolysis of 0. nervosum have shown a remarkable effect of the alkaline pretreatments on the final hydrolysis yield. (Author) 10 refs.

  14. Effects of grinding processes on enzymatic degradation of wheat straw. (United States)

    Silva, Gabriela Ghizzi D; Couturier, Marie; Berrin, Jean-Guy; Buléon, Alain; Rouau, Xavier


    The effectiveness of wheat straw fine to ultra-fine grindings at pilot scale was studied. The produced powders were characterised by their particle-size distribution (laser diffraction), crystallinity (WAXS) and enzymatic degradability (Trichoderma reesei enzymatic cocktail). A large range of wheat-straw powders was produced: from coarse (median particle size ∼800 μm) to fine particles (∼50 μm) using sieve-based grindings, then ultra-fine particles ∼20 μm by jet milling and ∼10 μm by ball milling. The wheat straw degradability was enhanced by the decrease of particle size until a limit: ∼100 μm, up to 36% total carbohydrate and 40% glucose hydrolysis yields. Ball milling samples overcame this limit up to 46% total carbohydrate and 72% glucose yields as a consequence of cellulose crystallinity reduction (from 22% to 13%). Ball milling appeared to be an effective pretreatment with similar glucose yield and superior carbohydrate yield compared to steam explosion pretreatment.

  15. Novel investigation of enzymatic biodiesel reaction by isothermal calorimetry

    Energy Technology Data Exchange (ETDEWEB)

    Sotoft, Lene Fjerbaek, E-mail: [Institute of Chemical Engineering, Biotechnology and Environmental Technology, University of Southern Denmark, Campusvej 55, DK-5230 Odense M (Denmark); Westh, Peter [Department of Life Science and Chemistry, Roskilde University, PO Box 260, DK-4000 Roskilde (Denmark); Christensen, Knud V.; Norddahl, Birgir [Institute of Chemical Engineering, Biotechnology and Environmental Technology, University of Southern Denmark, Campusvej 55, DK-5230 Odense M (Denmark)


    Isothermal calorimetry (ITC) was used to investigate solvent-free enzymatic biodiesel production. The transesterification of rapeseed oil with methanol and ethanol was catalyzed by immobilized lipase Novozym 435 at 40 {sup o}C. The aim of the study was to determine reaction enthalpy for the enzymatic transesterification and to elucidate the mass transfer and energetic processes taking place. Based on the measured enthalpy and composition change in the system, the heat of reaction at 40 {sup o}C for the two systems was determined as -9.8 {+-} 0.9 kJ/mole biodiesel formed from rapeseed oil and methanol, and -9.3 {+-} 0.7 kJ/mole when rapeseed oil and ethanol was used. Simple Michaelis-Menten kinetics was not an appropriate choice for describing the kinetics of this heterogeneous system. The experiments demonstrated the possibility of investigating complex reaction mixtures using ITC. Although it is possible to determine thermodynamic properties such as reaction enthalpy and reaction rate, the difficulty in actually measuring the true non-mass-transfer-limited reaction kinetics is exposed by the high time resolution of ITC.

  16. Process Evaluation Tools for Enzymatic Cascades Welcome Message

    DEFF Research Database (Denmark)

    Abu, Rohana

    Biocatalysis is attracting significant attention from both academic and industrial scientists due to the excellent capability of enzyme to catalyse selective reactions. Recently, much interest has been shown in the application of enzymatic cascades as a useful tool in organic synthesis to synthes......Biocatalysis is attracting significant attention from both academic and industrial scientists due to the excellent capability of enzyme to catalyse selective reactions. Recently, much interest has been shown in the application of enzymatic cascades as a useful tool in organic synthesis...... the atom economy. The scheme consists of two primary enzymes (alcohol dehydrogenase and ω-transaminase) that are directly involved in the main synthesis. Alanine dehydrogenase was introduced as a secondary enzyme to regenerate the co-factor NAD+ and co-substrate alanine in situ as well as to shift...... the equilibrium positions in the main syntheses. In principle, this strategy could successfully achieve high conversion, using ammonia as the sole reagent used in excess to drive the conversion. The findings herein indicate that quantitatively the possibilities for improving the conversion of thermodynamically...

  17. An enzymatic platform for the synthesis of isoprenoid precursors.

    Directory of Open Access Journals (Sweden)

    Sofia B Rodriguez

    Full Text Available The isoprenoid family of compounds is estimated to contain ∼ 65,000 unique structures including medicines, fragrances, and biofuels. Due to their structural complexity, many isoprenoids can only be obtained by extraction from natural sources, an inherently risky and costly process. Consequently, the biotechnology industry is attempting to genetically engineer microorganisms that can produce isoprenoid-based drugs and fuels on a commercial scale. Isoprenoid backbones are constructed from two, five-carbon building blocks, isopentenyl 5-pyrophosphate and dimethylallyl 5-pyrophosphate, which are end-products of either the mevalonate or non-mevalonate pathways. By linking the HMG-CoA reductase pathway (which produces mevalonate to the mevalonate pathway, these building block can be synthesized enzymatically from acetate, ATP, NAD(PH and CoA. Here, the enzymes in these pathways are used to produce pathway intermediates and end-products in single-pot reactions and in remarkably high yield, ∼ 85%. A strategy for the regio-specific incorporation of isotopes into isoprenoid backbones is developed and used to synthesize a series of isotopomers of diphosphomevalonate, the immediate end-product of the mevalonate pathway. The enzymatic system is shown to be robust and capable of producing quantities of product in aqueous solutions that meet or exceed the highest levels achieved using genetically engineered organisms in high-density fermentation.

  18. Disposable cuvette test for enzymatic determination of heavy metals

    Energy Technology Data Exchange (ETDEWEB)

    Wolfbeis, O.S.; Preininger, C. [Karl-Franzens Univ., Graz (Austria). Institute of Organic Chemistry


    The authors report on an optical cuvette test for total heavy metals based on the inhibition of the enzyme urease by metals ions including silver(I), mercury(II), copper(II), nickel(II), cobalt(II), and cadmium(II). The enzymatic action is monitored using an optical ammonia transducer deposited on the wall of a disposable cuvette. This results in a rapid and inexpensive single-shot device for heavy metal sensing. A solution of urease and buffer is placed in the cuvette with the ammonium sensor membrane fixed on one of its walls. Enzymatic action starts after addition of a defined quantity of urea. This is indicated by the increase in the absorption of the ammonia sensor membrane whose color changes from yellow to blue. The slope of the increase in signal is the information for the un-inhibited reaction. After several minutes, the sample (containing the heavy metal) is added to the cuvette. Heavy metal ions inhibit the enzyme (by binding to the sulfhydryl groups) and cause a decrease in the slope. The ratio of slopes of un-inhibited and inhibited reactions is a direct parameter for detection and calculation total heavy metals.

  19. Enzymatic activity of three sugarcane varieties under salt stress

    Directory of Open Access Journals (Sweden)

    Mercia F. Carvalho

    Full Text Available ABSTRACT Sugarcane is a species of great economic importance to Brazil. The study of enzymatic activity has helped to identify varieties of sugarcane tolerant to salt stress, for planting in Brazilian semiarid regions, where there is the occurrence of saline soils. The objective of this study was to evaluate the enzymatic activity of three varieties of sugarcane subjected to stress caused by salinity. The experiment was conducted in a greenhouse at the Federal Rural University of Pernambuco, in a completely randomized design with four replicates. The leaf blade samples were collected for analysis of soluble proteins and determination of the activity of catalase and ascorbate peroxidase. The protein content was higher for RB962962 and lower for the varieties RB92579 and RB867515. However, in RB867515, there was an increase in the activity of ascorbate peroxidase and catalase, while the varieties RB92579 and RB962962 showed a reduction in the activity of these enzymes. These results indicate that the variety RB867515 has a more efficient mechanism in plant protection against the action of reactive oxygen species.

  20. Production of intracellular enzymes by enzymatic treatment of yeast

    Energy Technology Data Exchange (ETDEWEB)

    Zomer, E.; Er-El, Z.; Rokem, J.S.


    Enzymatic extraction of intracellular enzymes from various yeasts by glucanase was investigated. Favourable conditions for lysis and release of intracellular enzymes were established. The effects of yeast concentration, growth phase of yeast, storage temperature and pretreatment of yeast were studied. The yeasts investigated can be divided into two groups. The first, Kluyveromyces lactis, Saccharomyces cerevisiae, Saccharomyces oviformis, Torulopsis glabrata, Hansenula polymorpha and local bakers' yeast, lysed relatively easily (70-80% of the cells), especially when cells from the logarithmic growth phase were treated. The second, Candida utilis and Candida vini, were more susceptible to lysis (40-50%) when cells were taken from the stationary phase. Release of two enzymes, glycerol kinase from Candida utilis grown on glycerol and formate dehydrogenase from Torulopsis glabrata grown on methanol was examined. The highest specific activities were obtained by incubating the cells with glucanase for 1.5 hours at 37 degrees C. Inactivation of the released enzyme was relatively low. After 12 hours of enzymatic treatment at 28 degrees C glycerol kinase maintained about 50%, and formate dehydrogenase over 80%, of the original activities. (Refs. 12).

  1. Synergism and Mutualism in Non-Enzymatic RNA Polymerization

    Directory of Open Access Journals (Sweden)

    Hussein Kaddour


    Full Text Available The link between non-enzymatic RNA polymerization and RNA self-replication is a key step towards the “RNA world” and still far from being solved, despite extensive research. Clay minerals, lipids and, more recently, peptides were found to catalyze the non-enzymatic synthesis of RNA oligomers. Herein, a review of the main models for the formation of the first RNA polymers is presented in such a way as to emphasize the cooperation between life’s building blocks in their emergence and evolution. A logical outcome of the previous results is a combination of these models, in which RNA polymerization might have been catalyzed cooperatively by clays, lipids and peptides in one multi-component prebiotic soup. The resulting RNAs and oligopeptides might have mutualistically evolved towards functional RNAs and catalytic peptides, preceding the first RNA replication, thus supporting an RNA-peptide world. The investigation of such a system is a formidable challenge, given its complexity deriving from a tremendously large number of reactants and innumerable products. A rudimentary experimental design is outlined, which could be used in an initial attempt to study a quaternary component system.

  2. Synergism and Mutualism in Non-Enzymatic RNA Polymerization (United States)

    Kaddour, Hussein; Sahai, Nita


    The link between non-enzymatic RNA polymerization and RNA self-replication is a key step towards the “RNA world” and still far from being solved, despite extensive research. Clay minerals, lipids and, more recently, peptides were found to catalyze the non-enzymatic synthesis of RNA oligomers. Herein, a review of the main models for the formation of the first RNA polymers is presented in such a way as to emphasize the cooperation between life’s building blocks in their emergence and evolution. A logical outcome of the previous results is a combination of these models, in which RNA polymerization might have been catalyzed cooperatively by clays, lipids and peptides in one multi-component prebiotic soup. The resulting RNAs and oligopeptides might have mutualistically evolved towards functional RNAs and catalytic peptides, preceding the first RNA replication, thus supporting an RNA-peptide world. The investigation of such a system is a formidable challenge, given its complexity deriving from a tremendously large number of reactants and innumerable products. A rudimentary experimental design is outlined, which could be used in an initial attempt to study a quaternary component system. PMID:25370531

  3. Improvement of Soybean Oil Solvent Extraction through Enzymatic Pretreatment

    Directory of Open Access Journals (Sweden)

    F. V. Grasso


    Full Text Available The purpose of this study is to evaluate multienzyme hydrolysis as a pretreatment option to improve soybean oil solvent extraction and its eventual adaptation to conventional processes. Enzymatic action causes the degradation of the cell structures that contain oil. Improvements in terms of extraction, yield, and extraction rate are expected to be achieved. Soybean flakes and collets were used as materials and hexane was used as a solvent. Temperature, pH, and incubation time were optimized and diffusion coefficients were estimated for each solid. Extractions were carried out in a column, oil content was determined according to time, and a mathematical model was developed to describe the system. The optimum conditions obtained were pH 5.4, 38°C, and 9.7 h, and pH 5.8, 44°C, and 5.8h of treatment for flakes and collets, respectively. Hydrolyzed solids exhibited a higher yield. Diffusion coefficients were estimated between 10-11 and 10-10. The highest diffusion coefficient was obtained for hydrolyzed collets. 0.73 g oil/mL and 0.7 g oil/mL were obtained at 240 s in a column for collets and flakes, respectively. Hydrolyzed solids exhibited a higher yield. The enzymatic incubation accelerates the extraction rate and allows for higher yield. The proposed model proved to be appropriate.

  4. Value addition of corn husks through enzymatic production of xylooligosaccharides

    Directory of Open Access Journals (Sweden)

    Ashis Kumar Samanta

    Full Text Available ABSTRACT Corn husks are the major wastes of corn industries with meagre economic significance. The present study was planned for value addition of corn husk through extraction of xylan, followed by its enzymatic hydrolysis into xylooligosaccharides, a pentose based prebiotic. Compositional analysis of corn husks revealed neutral detergent fibre 68.87%, acid detergent fibre 31.48%, hemicelluloses 37.39%, cellulose 29.07% and crude protein 2.68%. Irrespective of the extraction conditions, sodium hydroxide was found to be more effective in maximizing the yield of xylan from corn husks than potassium hydroxide (84% vs. 66%. Application of xylanase over the xylan of corn husks resulted into production of xylooligosaccharides with different degree of polymerization namely, xylobiose and xylotriose in addition to xylose monomer. On the basis of response surface model analysis, the maximum yield of xylobiose (1.9 mg/ml was achieved with the enzymatic hydrolysis conditions of pH 5.8, temperature 44°C, enzyme dose 5.7U/ml and hydrolysis time of 17.5h. Therefore, the corn husks could be used as raw material for xylan extraction vis a vis its translation into prebiotic xylooligosaccharides.

  5. Enzymatic hydrolysis: a method in alleviating legume allergenicity. (United States)

    Kasera, Ramkrashan; Singh, A B; Lavasa, S; Prasad, Komarla Nagendra; Arora, Naveen


    Legumes are involved in IgE mediated food allergy in many countries. Avoidance of allergenic food is the only way to avoid symptomatic reaction. The present study investigated the effect of enzymatic hydrolysis on the allergenicity of three legumes - kidney bean (Phaseolus vulgaris), black gram (Vigna mungo) and peanut (Arachis hypogaea). Soluble protein extracts of the study legumes were sequentially treated by Alcalase(®) and Flavourzyme(®). Allergenicity of hydrolysates was then determined by ELISA, immunoblot, stripped basophil histamine release and skin prick test (SPT). Hydrolysis resulted in the loss of all IgE binding fractions determined by immunoblot in the three legumes. Specific IgE binding in ELISA was reduced by 62.2 ± 7.7%, 87.1 ± 9.6% and 91.8 ± 7.2% in the hydrolysates of kidney bean, black gram and peanut, respectively (p release of histamine was decreased significantly when sensitized basophils were challenged with hydrolysates as compared to raw extracts. Significant reduction in the biopotency of hydrolysates was also observed in SPT where only 1/10 kidney bean-sensitive individuals, 2/6 black gram-sensitive individuals and 1/7 peanut-sensitive individuals were found positive to their respective hydrolysates. In conclusion, enzymatic hydrolysis is effective in attenuating allergenicity of legume proteins and may be employed for preparing hypoallergenic food extracts.

  6. Enzymatic hydrolysis of fructans in the tequila production process. (United States)

    Avila-Fernández, Angela; Rendón-Poujol, Xóchitl; Olvera, Clarita; González, Fernando; Capella, Santiago; Peña-Alvarez, Araceli; López-Munguía, Agustín


    In contrast to the hydrolysis of reserve carbohydrates in most plant-derived alcoholic beverage processes carried out with enzymes, agave fructans in tequila production have traditionally been transformed to fermentable sugars through acid thermal hydrolysis. Experiments at the bench scale demonstrated that the extraction and hydrolysis of agave fructans can be carried out continuously using commercial inulinases in a countercurrent extraction process with shredded agave fibers. Difficulties in the temperature control of large extraction diffusers did not allow the scaling up of this procedure. Nevertheless, batch enzymatic hydrolysis of agave extracts obtained in diffusers operating at 60 and 90 degrees C was studied at the laboratory and industrial levels. The effects of the enzymatic process on some tequila congeners were studied, demonstrating that although a short thermal treatment is essential for the development of tequila's organoleptic characteristics, the fructan hydrolysis can be performed with enzymes without major modifications in the flavor or aroma, as determined by a plant sensory panel and corroborated by the analysis of tequila congeners.

  7. A Minimalist Substrate for Enzymatic Peptide and Protein Conjugation (United States)

    Wollack, James W.; Silverman, Julie M.; Petzold, Christopher J.; Mougous, Joseph D.; Distefano, Mark D.


    Recently a number of non-natural prenyl groups containing alkynes and azides have been developed as handles to perform click chemistry on proteins and peptides ending in the sequence “CAAX”, where C is a cysteine that becomes alkylated, A is an aliphatic amino acid and X is any amino acid. When such molecules are modified, a tag containing a prenyl analog and the “CAAX box” sequence remains. Here we report the synthesis of an alkyne-containing substrate comprised of only nine non-hydrogen atoms. This substrate was synthesized in six steps from 3-methyl-2-buten-1-ol and has been enzymatically incorporated into both proteins and peptides using protein farnesyltransferase. After prenylation the final three amino acids required for enzymatic recognition can be removed using carboxypeptidase Y, leaving a single residue (the cysteine from the “CAAX box”) and the prenyl analog as the only modifications. We also demonstrate that this small tag minimizes the impact of the modification on the solubility of the targeted protein. Hence, this new approach should be useful for applications in which the presence of a large tag hinders the modified protein's solubility, reactivity or utility. PMID:19856367

  8. Highly Efficient Enzymatic Preparation of Daidzein in Deep Eutectic Solvents

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    Qi-Bin Cheng


    Full Text Available Daidzein, which is scarce in nature, has gained significant attention due to its superior biological activity and bioavailability compared with daidzin. So far, it has been widely used in the medicine and health care products industries. The enzymatic approach for the preparation of daidzein has prevailed, benefitted by its high efficiency and eco-friendly nature. Our present research aimed at providing a preparation method of daidzein by enzymatic hydrolysis of daidzin in a new “green” reaction medium-deep eutectic solvents (DESs. Herein, the DESs were screened via evaluating enzyme activity, enzyme stability and the substrate solubility, and the DES (ChCl/EG 2:1, 30 vol % was believed to be the most appropriate co-solvent to improve the bioconversion efficiency. Based on the yield of daidzein, response surface methodology (RSM was employed to model and optimize the reaction parameters. Under these optimum process conditions, the maximum yield of 97.53% was achieved and the purity of daidzein crude product reached more than 70%, which is more efficient than conversions in DESs-free buffer. Importantly, it has been shown that DESs medium could be reused for six batches of the process with a final conversion of above 50%. The results indicated that this procedure could be considered a mild, environmentally friendly, highly efficient approach to the economical production of daidzein, with a simple operation process and without any harmful reagents being involved.

  9. Chemical and Enzymatic Hydrolysis of Polyurethane/Polylactide Blends

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    Joanna Brzeska


    Full Text Available Polyether-esterurethanes containing synthetic poly[(R,S-3-hydroxybutyrate] (R,S-PHB and polyoxytetramethylenediol in soft segments and polyesterurethanes with poly(ε-caprolactone and poly[(R,S-3-hydroxybutyrate] were blended with poly([D,L]-lactide (PLA. The products were tested in terms of their oil and water absorption. Oil sorption tests of polyether-esterurethane revealed their higher response in comparison to polyesterurethanes. Blending of polyether-esterurethanes with PLA caused the increase of oil sorption. The highest water sorption was observed for blends of polyether-esterurethane, obtained with 10% of R,S-PHB in soft segments. The samples mass of polyurethanes and their blends were almost not changed after incubation in phosphate buffer and trypsin and lipase solutions. Nevertheless the molecular weight of polymers was significantly reduced after degradation. It was especially visible in case of incubation of samples in phosphate buffer what suggested the chemical hydrolysis of polymer chains. The changes of surface of polyurethanes and their blends, after incubation in both enzymatic solutions, indicated on enzymatic degradation, which had been started despite the lack of mass lost. Polyurethanes and their blends, contained more R,S-PHB in soft segments, were degraded faster.

  10. A characterization of scale invariant responses in enzymatic networks.

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    Maja Skataric

    Full Text Available An ubiquitous property of biological sensory systems is adaptation: a step increase in stimulus triggers an initial change in a biochemical or physiological response, followed by a more gradual relaxation toward a basal, pre-stimulus level. Adaptation helps maintain essential variables within acceptable bounds and allows organisms to readjust themselves to an optimum and non-saturating sensitivity range when faced with a prolonged change in their environment. Recently, it was shown theoretically and experimentally that many adapting systems, both at the organism and single-cell level, enjoy a remarkable additional feature: scale invariance, meaning that the initial, transient behavior remains (approximately the same even when the background signal level is scaled. In this work, we set out to investigate under what conditions a broadly used model of biochemical enzymatic networks will exhibit scale-invariant behavior. An exhaustive computational study led us to discover a new property of surprising simplicity and generality, uniform linearizations with fast output (ULFO, whose validity we show is both necessary and sufficient for scale invariance of three-node enzymatic networks (and sufficient for any number of nodes. Based on this study, we go on to develop a mathematical explanation of how ULFO results in scale invariance. Our work provides a surprisingly consistent, simple, and general framework for understanding this phenomenon, and results in concrete experimental predictions.

  11. Experiencing affective interactive art

    NARCIS (Netherlands)

    Bialoskorski, Leticia S.S.; Westerink, Joyce H.D.M.; Broek, van den Egon L.


    The progress in the field of affective computing enables the realization of affective art. This paper describes the affective interactive art system Mood Swings, which interprets and visualizes affect expressed by a person. Mood Swings is founded on the integration of a framework for affective move

  12. The Rapid and Sensitive Quantitative Determination of Galactose by Combined Enzymatic and Colorimetric Method: Application in Neonatal Screening. (United States)

    Kianmehr, Anvarsadat; Mahrooz, Abdolkarim; Ansari, Javad; Oladnabi, Morteza; Shahbazmohammadi, Hamid


    The quantitative measurement of galactose in blood is essential for the early diagnosis, treatment, and dietary monitoring of galactosemia patients. In this communication, we aimed to develop a rapid, sensitive, and cost-effective combined method for galactose determination in dry blood spots. This procedure was based on the combination of enzymatic reactions of galactose dehydrogenase (GalDH), dihydrolipoyl dehydrogenase (DLD), and alkaline phosphates with a colorimetric system. The incubation time and the concentration of enzymes used in new method were also optimized. The analytical performance was studied by the precision, recovery, linearity, and sensitivity parameters. Statistical analysis was applied to method comparison experiment. The regression equation and correlation coefficient (R (2)) were Y = 0.0085x + 0.032 and R (2) = 0.998, respectively. This assay exhibited a recovery in the range of 91.7-114.3 % and had the limit detection of 0.5 mg/dl for galactose. The between-run coefficient of variation (CV) was between 2.6 and 11.1 %. The within-run CV was between 4.9 and 9.2 %. Our results indicated that the new and reference methods were in agreement because no significant biases exist between them. Briefly, a quick and reliable combined enzymatic and colorimetric assay was presented for application in newborn mass screening and monitoring of galactosemia patients.

  13. Microbial/enzymatic synthesis of chiral drug intermediates. (United States)

    Patel, R N


    Biocatalytic processes were used to prepare chiral intermediates for pharmaceuticals. These include the following processes. Enzymatic synthesis of [4S-(4a,7a,10ab)]1-octahydro-5-oxo-4-[[(phenylmethoxy) carbonyl]amino]-7H-pyrido-[2,1-b] [1,3]thiazepine-7-carboxylic acid methyl ester (BMS-199541-01), a key chiral intermediate for synthesis of a new vasopeptidase inhibitor. Enzymatic oxidation of the epsilon-amino group of lysine in dipeptide dimer N2-[N[[(phenylmethoxy)carbonyl] L-homocysteinyl] L-lysine)1,1-disulfide (BMS-201391-01) to produce BMS-199541-01 using a novel L-lysine epsilon-aminotransferase from S. paucimobilis SC16113 was demonstrated. This enzyme was overexpressed in E. coli, and a process was developed using recombinant enzyme. The aminotransferase reaction required alpha-ketoglutarate as the amine acceptor. Glutamate formed during this reaction was recycled back to alpha-ketoglutarate by glutamate oxidase from S. noursei SC6007. Synthesis and enzymatic conversion of 2-keto-6-hydroxyhexanoic acid 5 to L-6-hydroxy norleucine 4 was demonstrated by reductive amination using beef liver glutamate dehydrogenase. To avoid the lengthy chemical synthesis of ketoacid 5, a second route was developed to prepare the ketoacid by treatment of racemic 6-hydroxy norleucine (readily available from hydrolysis of 5-(4-hydroxybutyl) hydantoin, 6) with D-amino acid oxidase from porcine kidney or T. variabilis followed by reductive amination to convert the mixture to L-6-hydroxynorleucine in 98% yield and 99% enantiomeric excess. Enzymatic synthesis of (S)-2-amino-5-(1,3-dioxolan-2-yl)-pentanoic acid (allysine ethylene acetal, 7), one of three building blocks used for synthesis of a vasopeptidase inhibitor, was demonstrated using phenylalanine dehydrogenase from T. intermedius. The reaction requires ammonia and NADH. NAD produced during the reaction was recycled to NADH by oxidation of formate to CO2 using formate dehydrogenase. Efficient synthesis of chiral

  14. Regenerating cellulose from ionic liquids for an accelerated enzymatic hydrolysis

    Energy Technology Data Exchange (ETDEWEB)

    Zhao, Hua [Savannah State University; Jones, Cecil L [Savannah State University; Baker, Gary A [ORNL; Xia, Shuqian [Tianjin University, Tianjin, China; Olubajo, Olarongbe [Savannah State University; Person, Vernecia [Savannah State University


    The efficient conversion of lignocellulosic materials into fuel ethanol has become a research priority in producing affordable and renewable energy. The pretreatment of lignocelluloses is known to be key to the fast enzymatic hydrolysis of cellulose. Recently, certain ionic liquids (ILs)were found capable of dissolving more than 10 wt% cellulose. Preliminary investigations [Dadi, A.P., Varanasi, S., Schall, C.A., 2006. Enhancement of cellulose saccharification kinetics using an ionic liquid pretreatment step. Biotechnol. Bioeng. 95, 904 910; Liu, L., Chen, H., 2006. Enzymatic hydrolysis of cellulose materials treated with ionic liquid [BMIM]Cl. Chin. Sci. Bull. 51, 2432 2436; Dadi, A.P., Schall, C.A., Varanasi, S., 2007. Mitigation of cellulose recalcitrance to enzymatic hydrolysis by ionic liquid pretreatment. Appl. Biochem. Biotechnol. 137 140, 407 421] suggest that celluloses regenerated from IL solutions are subject to faster saccharification than untreated substrates. These encouraging results offer the possibility of using ILs as alternative and nonvolatile solvents for cellulose pretreatment. However, these studies are limited to two chloride-based ILs: (a) 1-butyl-3-methylimidazolium chloride ([BMIM]Cl), which is a corrosive, toxic and extremely hygroscopic solid (m.p. 70 C), and (b) 1-allyl-3-methylimidazolium chloride ([AMIM]Cl), which is viscous and has a reactive side-chain. Therefore, more in-depth research involving other ILs is much needed to explore this promising pretreatment route. For this reason, we studied a number of chloride- and acetate-based ILs for cellulose regeneration, including several ILs newly developed in our laboratory. This will enable us to select inexpensive, efficient and environmentally benign solvents for processing cellulosic biomass. Our data confirm that all regenerated celluloses are less crystalline (58 75% lower) and more accessible to cellulase (>2 times) than untreated substrates. As a result, regenerated Avicel

  15. Enzymatic corn wet milling: engineering process and cost model

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    McAloon Andrew J


    Full Text Available Abstract Background Enzymatic corn wet milling (E-milling is a process derived from conventional wet milling for the recovery and purification of starch and co-products using proteases to eliminate the need for sulfites and decrease the steeping time. In 2006, the total starch production in USA by conventional wet milling equaled 23 billion kilograms, including modified starches and starches used for sweeteners and ethanol production 1. Process engineering and cost models for an E-milling process have been developed for a processing plant with a capacity of 2.54 million kg of corn per day (100,000 bu/day. These models are based on the previously published models for a traditional wet milling plant with the same capacity. The E-milling process includes grain cleaning, pretreatment, enzymatic treatment, germ separation and recovery, fiber separation and recovery, gluten separation and recovery and starch separation. Information for the development of the conventional models was obtained from a variety of technical sources including commercial wet milling companies, industry experts and equipment suppliers. Additional information for the present models was obtained from our own experience with the development of the E-milling process and trials in the laboratory and at the pilot plant scale. The models were developed using process and cost simulation software (SuperPro Designer® and include processing information such as composition and flow rates of the various process streams, descriptions of the various unit operations and detailed breakdowns of the operating and capital cost of the facility. Results Based on the information from the model, we can estimate the cost of production per kilogram of starch using the input prices for corn, enzyme and other wet milling co-products. The work presented here describes the E-milling process and compares the process, the operation and costs with the conventional process. Conclusion The E-milling process

  16. Unraveling the Enzymatic Basis of Wine “Flavorome”: A Phylo-Functional Study of Wine Related Yeast Species (United States)

    Belda, Ignacio; Ruiz, Javier; Alastruey-Izquierdo, Ana; Navascués, Eva; Marquina, Domingo; Santos, Antonio


    Non-Saccharomyces yeasts are a heterogeneous microbial group involved in the early stages of wine fermentation. The high enzymatic potential of these yeasts makes them a useful tool for increasing the final organoleptic characteristics of wines in spite of their low fermentative power. Their physiology and contribution to wine quality are still poorly understood, with most current knowledge being acquired empirically and in most cases based in single species and strains. This work analyzed the metabolic potential of 770 yeast isolates from different enological origins and representing 15 different species, by studying their production of enzymes of enological interest and linking phylogenetic and enzymatic data. The isolates were screened for glycosidase enzymes related to terpene aroma release, the β-lyase activity responsible for the release of volatile thiols, and sulfite reductase. Apart from these aroma-related activities, protease, polygalacturonase and cellulase activities were also studied in the entire yeast collection, being related to the improvement of different technological and sensorial features of wines. In this context, and in terms of abundance, two different groups were established, with α-L-arabinofuranosidase, polygalacturonase and cellulase being the less abundant activities. By contrast, β-glucosidase and protease activities were widespread in the yeast collection studied. A classical phylogenetic study involving the partial sequencing of 26S rDNA was conducted in conjunction with the enzymatic profiles of the 770 yeast isolates for further typing, complementing the phylogenetic relationships established by using 26S rDNA. This has rendered it possible to foresee the contribution different yeast species make to wine quality and their potential applicability as pure inocula, establishing species-specific behavior. These consistent results allowed us to design future targeted studies on the impact different non-Saccharomyces yeast species

  17. Oligoglucans as Elicitors of an Enzymatic Antioxidant System in Zucchini Squash (Cucurbita pepo L. Seedlings at Low Temperature

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    Abel Ceron-Garcia


    Full Text Available Problem statement: Non-controlled low temperatures affect physiological and biochemical processes leading to Reactive Oxygen Species (ROS accumulation in plants, which normally are stabilized by the antioxidant system. Superoxide Dismutase (SOD, Ascorbate Peroxidase (APX and Catalase (CAT are the most important ROS-detoxification enzymes in stressed plants. Approach: The activity of these enzymes were examined in response to fungal Oligoglucans (OG in cold-stressed cotyledons of Cucurbita pepo L. Cold stress condition was settled in zucchini cotyledons using relative Ion Leakage (IL and H2O2 accumulation. Then, OG were obtained from Trichoderma harzianum cellwall by enzymatic and chemical hydrolysis. These elicitors were purified by size-exclusion and anionexchange chromatography and tested in cold-stressed cotyledons. Results: No significant changes on H2O2 content and IL were observed between treated and control cold-stressed cotyledons, during the first 7 days of storage at 5°C. Afterwards H2O2 values increased greatly after cold-stressed. Then, cold stress conditions were established (7 d, 5°C. The APX and CAT activities remained stable, but not SOD activity, which decreased about 42% in control cotyledons. When a chemical OG mixture (CM was applied to cold-stressed cotyledons, APX and CAT activity levels increased. The SOD activity was only increased by an enzymatic OG mixture (EM. The APX activity level was increased in coldstressed cotyledons using OG size-exclusion chromatography fractions from both CM and EM; however, changes in CAT activity were only possible using EM fractions and no changes were detected in SOD activity. The highest CAT activity levels were triggered using OG anion-exchange chromatography sub-fractions from both CM and EM fractions (120 and 119 % of enzymatic activity, respectively. Conclusion: The oxidative stress can be counteracted by the triggering of APX and CAT activity in cold

  18. Evaluation of relationships between growth rate, tree size, lignocellulose composition and enzymatic saccharification in interspecific Corymbia hybrids and parental taxa.

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    Adam L Healey


    Full Text Available In order for a lignocellulosic bioenergy feedstock to be considered sustainable, it must possess a high rate of growth to supply biomass for conversion. Despite the desirability of a fast growth rate for industrial application, it is unclear what effect growth rate has on biomass composition or saccharification. We characterized Klason lignin, glucan, and xylan content with response to growth in Corymbia interspecific F1 hybrid families (HF and parental species C. torelliana (CT and C. citriodora subspecies variegata (CCV and measured the effects on enzymatic hydrolysis from hydrothermally pretreated biomass. Analysis of biomass composition within Corymbia populations found similar amounts of Klason lignin content (19.7-21.3% among parental and hybrid populations, whereas glucan content was clearly distinguished within CCV (52% and HF148 (60% as compared to other populations (28-38%. Multiple linear regression indicates that biomass composition is significantly impacted by tree size measured at the same age, with Klason lignin content increasing with diameter breast height (DBH (+0.12% per cm DBH increase, and glucan and xylan typically decreasing per DBH cm increase (-0.7% and -0.3%, respectively. Polysaccharide content within CCV and HF-148 were not significantly affected by tree size. High-throughput enzymatic saccharification of hydrothermally pretreated biomass found significant differences among Corymbia populations for total glucose production from biomass, with parental CT and hybrids HF-148 and HF-51 generating the highest amounts of glucose (~180 mg/g biomass, respectively, with HF-51 undergoing the most efficient glucan-to-glucose conversion (74%. Based on growth rate, biomass composition, and further optimization of enzymatic saccharification yield, high production Corymbia hybrid trees are potentially suitable for fast-rotation bioenergy or biomaterial production.

  19. Improved QM Methods and Their Application in QM/MM Studies of Enzymatic Reactions (United States)

    Jorgensen, William L.


    Quantum mechanics (QM) and Monte Carlo statistical mechanics (MC) simulations have been used by us since the early 1980s to study reaction mechanisms and the origin of solvent effects on reaction rates. A goal was always to perform the QM and MC/MM calculations simultaneously in order to obtain free-energy surfaces in solution with no geometrical restrictions. This was achieved by 2002 and complete free-energy profiles and surfaces with full sampling of solute and solvent coordinates can now be obtained through one job submission using BOSS [JCC 2005, 26, 1689]. Speed and accuracy demands also led to development of the improved semiempirical QM method, PDDG-PM3 [JCC 1601 (2002); JCTC 817 (2005)]. The combined PDDG-PM3/MC/FEP methodology has provided excellent results for free energies of activation for many reactions in numerous solvents. Recent examples include Cope, Kemp and E1cb eliminations [JACS 8829 (2005), 6141 (2006); JOC 4896 (2006)], as well as enzymatic reactions catalyzed by the putative Diels-Alderase, macrophomate synthase, and fatty-acid amide hydrolase [JACS 3577 (2005); JACS (2006)]. The presentation will focus on the accuracy that is currently achievable in such QM/MM studies and the accuracy of the underlying QM methodology including extensive comparisons of results from PDDG-PM3 and ab initio DFT methods.

  20. Enzymatic Resolution and Separation of Secondary Alcohols Based on Fatty Esters as Acylating Agents (United States)

    Monteiro, Carlos M.; Afonso, Carlos A. M.; Lourenco, Nuno M. T.


    The enzymatic resolution of "rac"-1-phenylethanol using ethyl myristate as acylating agent and solvent and "Candida antarctica" lipase B (CAL-B) as biocatalyst was demonstrated with catalyst and medium reuse. Both enantiomers of 1-phenylethanol were isolated by sequential enzymatic reactions and product distillations. From the first enzymatic…