Orientador: Adriana Zerlotti Mercadante / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-13T03:20:52Z (GMT). No. of bitstreams: 1 Barbosa_MariaIvoneMartinsJacintho_D.pdf: 1261091 bytes, checksum: 8a753aaea026de68ffab9783fd8d3cfb (MD5) Previous issue date: 2009 / Resumo: A bixina foi extraída de sementes de urucum e purificada através de sucessivas cristalizações. A seguir a bixina foi encapsulada por atomização, utilizando-se como materiais de parede diferentes misturas com goma arábica (GA), maltodextrina 20 dextrose equivalente (MD) e sacarose (Sac), com ou sem o emulsificante Tween 80. Todas as soluções de materiais de parede foram preparadas com 40% de sólidos totais em ¿spraydryer¿ operando a 180º C e diâmetro de bico injetor de 0,07 mm. As características de todas as microcápsulas foram avaliadas e a estabilidade das microcápsulas de bixina foi estudada em sistemas aquoso, gel e sólido (maria-mole), expostos a diferentes intensidades de luz. Na primeira etapa deste trabalho foram testados os seguintes materiais de parede: 95% GA+ 5% Sac, 100% maltodextrina, 99,8% maltodextrina + 0,2% Tween 80, e 80% maltodextrina+ 20% Sac para a microencapsulação de bixina. Foram determinados os seguintes parâmetros: eficiência da microencapsulação, morfologia, solubilidade das microcápsulas de bixina, além da estabilidade em sistema-modelo aquoso sob luz (700 lux) e no escuro, a 21º C na presença de ar. As microcápsulas elaboradas com 95% GA + 5% Sac ou 99,8 % maltodextrina + 0,2 % Tween 80 foram as que apresentaram melhor eficiência da microencapsulação, 86 e 75%, respectivamente e, além disso, conferiram maior estabilidade a bixina durante a exposição à luz ou na estocagem no escuro. Em todos os sistemas encapsulados expostos à luz, a bixina apresentou duas etapas de degradação, que se ajustaram a cinética de primeira ordem. Na etapa seguinte, misturas de maltodextrina e goma arábica nas proporções de 1:1 e 4:1 foram utilizadas como materiais de parede para microencapsulação de bixina. A viscosidade das soluções dos materiais de parede foi determinada, bem como as seguintes propriedades das microcápsulas: retenção após secagem, rendimento, eficiência da microencapsulação e morfologia. Foi verificado que a solução de MD/GA1:1, de maior viscosidade (38,72 ± 0,26 cP), originou microcápsulas com maiores valores de retenção de bixina, rendimento de pó e eficiência da microencapsulação em comparação com a solução menos viscosa de MD/GA4:1. Apesar das diferenças observadas, apenas o rendimento e a eficiência do microencapsulação foram significativamente diferentes (p _ 0,05). A fim de se testar a estabilidade da bixina em um sistema sólido, a degradação foi avaliada em gel exposto na presença e ausência de luminosidade, ambos na presença de ar. Neste sistema, as microcápsulas obtidas com MD/GA1:1 foram as que conferiram maior estabilidade à bixina na presença e ausência de luminosidade. No gel, a cinética de degradação da bixina microencapsulada foi de ordem zero, sendo que foram observadas três etapas de degradação, enquanto que degradação da bixina não encapsulada ocorreu em apenas uma etapa. Conforme o esperado, a bixina encapsulada foi mais estável que a não encapsulada, com valores de kobs aproximadamente 13 a 15 vezes menores em comparação aos respectivos sistemas com bixina não encapsulada. As bixina encapsulada com MD:GA na proporção 1:1 foi utilizada como corante de maria-mole e a alteração da cor instrumental (sistema CIELab) foi avaliada sob 1300 lux e no escuro. Visualmente, a maria-mole colorida com bixina encapsulada apresentou alaranjado mais intenso do que a colorida com bixina não encapsulado. Como esperado, a luz apresentou um efeito deletério na coloração da maria-mole, verificado pela gradual diminuição das coordenadas de cor a* e b*, sendo mais intenso na colorida com bixina livre do que na colorida com microcápsulas de bixina. Não foram observadas alterações do L*, a* e b* na maria-mole estocada no escuro durante 130 horas. Na última etapa deste estudo foi avaliado o efeito da adição de um antioxidante natural, ácido ascórbico (AA), nas propriedades e estabilidade da bixina encapsulada com MD:GA (1:1) em gel exposto a diferentes luminosidades (0, 1300 e 2100 lux), na presença de ar a 21 ± 2º C. Como controle foi utilizado gel colorido com bixina não encapsulada acrescido de AA. Além do teor de bixina, paralelamente também foi avaliada instrumentalmente a perda de cor das amostras utilizando-se o sistema CIELab. Conforme o esperado a luminosidade teve um efeito deletério na estabilidade da bixina, refletido na diminuição dos parâmetros a*, b* e C*. A cinética de degradação da bixina foi de ordem zero, sendo que nos géis coloridos com microcápsulas foram observadas três etapas de fotodegradação, enquanto que nos coloridos com bixina não encapsulada foi observada apenas uma etapa. No escuro, a degradação da bixina também apresentou apenas uma etapa. Por outro lado, os teores de AA praticamente não sofreram alteração na presença de luz / Abstract: Bixin was extracted from annatto seeds and purified through cristalizations followed by encapsulation by spray-dryer. The wall materials used were different blends of gum Arabic (GA), maltodextrin 20 dextrose equivalent (MD) and sucrose (Suc), with and without emulsifier Tween 80. All the wall materials solutions were prepared with 40% total solids in a spray-dryer operating at 180º C with aspersion nozzle diameter of 0.07 mm. The characteristics of all microcapsules were evaluated and the stability of bixin microcapsules was monitored in model-systems, either in water, in gel or solid ("maria-mole"), exposed to different light intensities. In the first stage of the present study, the following wall materials were tested for bixin microencapsulation: 95% GA+ 5% Suc, 100% maltodextrin, 99.8% maltodextrin+0.2% Tween 80, and 80% maltodextrin+20% SUc. The microencapsulation efficiency and morfology were evaluated, along with the stability of the bixin microcapsules in an aqueous model-system under light (700 lux) and in the dark, both at 21º C in the presence of air. The microcapsules containing 95% GA+ 5% Suc and 99,8 % maltodextrin+0.2 % Tween 80 showed the highest encapsulation efficiency, respectively 86% and 75%, less superficial imperfections and higher stability under ligt and in the dark as compared to bixin encapsulated with maltodextrin alone or blended with sucrose. The kinetic behavior of bixin photodegradation in all encapsulated systems was composed by two first-order decays. In the next stage, blends of maltodextrin and gum Arabic at 1:1 and 4:1 proportions were used as wall materials for bixin microencapsulation. The viscosity of the wall material solutions was determined, as well as the following microcapsules properties: retention after dying, yield, microencapsulation efficiency and morfology. Bixin degradation was evaluated in gel exposed to the presence and absence of light, both under air. The MD/GA1:1 solution presented the highest viscosity (38.72 ± 0.26 cP), furnishing microcapsules with higher values of bixin retention, powder yield and microencapsulation efficiency as compared to the MD/GA4:1 solution with lower viscosity. Although these differences were observed, only the yield and microencapsulation efficiency were significatively different (p _ 005). Bixin microcapsulated with MD/GA1:1 was more stable under light or dark in the gel model-system. The degradation of both microencapsulated bixin followed zero-order kinetics, with three degradation steps, whilst not encapsulated bixin showed one degradation step. As expected, microencapsulated bixin was more stable than that not encapsulated, with kobs values ca. 13 to 15 times lower as compared to the correspondent systems with not encapsulated bixin. Bixin encapsulated with MD:GA (1:1) was added as colorant in "maria-mole" and color parameters (CIELab) changes were evaluated under 1300 lux and in the dark. Visually, "maria-mole" samples colored with bixin microcapsules showed a more intense orange color as compared to maria-mole colored with not encapsulated bixin. As expected, the color parameters a* and b* decreased under light, and this decrease was higher in "maria-mole" colored with not encapsulated bixin. No changes in L*, a* and b* values were observed in "maria-mole" stored in the dark during 130 hours. In the last stage, the effect of addition of a natural antioxidant, ascorbic acid (AA), on the microcapsules properties and bixin stability microencapsulated with MD:GA (1:1) was evaluated in a gel system exposed to different light intensities (0, 1300 e 2100 lux) at 21 ± 2º C in the presence of air. Bixin+AA, both not encapsulated, were used as control. Besides the levels of bixin, color was measured in a spetrocolorimeter using the CIElab system to evaluate the changes in the gel-systems. As expected, light was the main factor that caused loss of bixin, which was reflected on the decrease of the a*, b* and C * color parameters. The bixin degradation followed zero order kinetics, with three fotodegradation steps in the systems colored with both microcapsules, whilst not encapsulated bixin+AA showed one degradation step. In the dark, bixin degradation occurred in only one kinetic step. On the other hand, the AA levels remained practically unchanged in the presence and absence of light / Doutorado / Doutor em Ciência de Alimentos

Orientador: Rosiane Lopes Cunha / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-13T14:14:55Z (GMT). No. of bitstreams: 1 Sato_AnaCarlaKawazoe_D.pdf: 2187954 bytes, checksum: 9f67420e2e9bb6fdb40e51d15be43b4e (MD5) Previous issue date: 2009 / Resumo: A influência das características físicas e químicas do meio dispersante sobre a reologia de suspensões-modelo foi avaliada visando o entendimento do comportamento de sistemas reais como as polpas de frutas. Para isto, o trabalho foi dividido em duas etapas: 1) estudo do efeito da composição de sistemas-modelo de soro de polpas de frutas no comportamento reológico, e 2) estudo de suspensões-modelo de diferentes partículas dispersas em matrizes com diferentes propriedades físicas e químicas. Na primeira etapa, foi verificado o efeito da presença de pectinas de alto e baixo grau de metoxilação, assim como de suas misturas. Os resultados mostraram um efeito sinergístico da mistura destes diferentes tipos de pectinas, devido à maior viscosidade e pseudoplasticidade dos sistemas mistos quando comparados aos sistemas com um único tipo de pectina. Assim, ressaltou-se a importância dos diferentes tipos de ligação no comportamento reológico destes sistemas. A segunda etapa permitiu a compreensão das interações entre partículas e partícula-solvente e a sua influência no comportamento reológico de suspensões-modelo. De um modo geral, as interações entre partículas e partícula-solvente foram maiores nos meios menos viscosos e polares. Tais resultados foram refletidos pelo valor ajustado da fração máxima de empacotamento (fm) por diferentes modelos, dado que foi possível a adição de maior quantidade de sólidos para os sistemas com menos interações. Xantana foi adicionada às matrizes dispersantes para promover pseudoplasticidade, indicando comportamentos distintos quando adicionadas em água pura e em soluções aquosas de glicerina. No primeiro caso, as matrizes apresentaram comportamento altamente pseudoplástico, enquanto que a adição de mesma quantidade de xantana nas soluções aquosas de glicerina levou a um grande aumento na viscosidade, com menor influência no valor do índice de comportamento de escoamento. A adição de sólidos nestes dois tipos de matrizes levou a comportamentos distintos. No primeiro caso verificou-se uma leve desestruturação do sistema, refletido principalmente pela maior influência do módulo complexo com a freqüência e menor variação na viscosidade relativa das suspensões. Por outro lado, o aumento na fração de sólidos nos sistemas compostos com xantana, glicerina e água levou a suspensões mais estruturadas. Assim, os resultados apresentados mostram que além das características das partículas, as propriedades químicas e reológicas da matriz dispersante são essenciais na determinação do comportamento estrutural e de escoamento de suspensões e portanto, não deveriam ser desprezadas pelos modelos existentes / Abstract: The influence of the physical and chemical properties of dispersing matrices on the rheological behavior of model-suspensions was evaluated, aiming the understanding of real suspensions such as fruit pulps. The present research was divided into two parts: 1) influence of composition of serum of fruit pulps on their rheological characteristics, through the study of model-systems, and 2) evaluation of model-suspensions of different particles dispersed into matrices with distinct characteristics (physical and chemical). In the first part, the effects of low and high methoxy pectins were evaluated, as well as their mixture. Results indicated a synergistic effect on the mixture of different sorts of pectin, which was observed by the higher viscosity and pseudoplasticity of mixed systems as compared to pure pectin ones. Such results allowed emphasizing the importance of different binding characteristics on the rheological properties of these systems. The second part allowed for the comprehension of interparticle and particle-matrix interactions, and their influence on the rheological behavior of model suspensions. Generally, interactions were more pronounced at low viscous and polar matrices, which reflected the lower maximum packing fraction (fm) fitted by different models. Xanthan was added to dispersing matrices to impart pseudoplasticity, showing distinct behaviors when mixed in pure water and aqueous glycerin solutions. In the first case, matrices showed highly shear thinning behavior, while the addition of the same amount of xanthan in glycerin aqueous solutions promoted a significant increase on viscosity, with lower influence on the flow index behavior (n). Thus, distinct behavior was observed when increasing solids concentration on these two matrices. In the first case, the increase of the dependency of complex modulus with frequency and the lower influence of the relative viscosity reflected a slight loss of structure with solids addition. On the other hand, increasing solids content on systems composed with xanthan, glycerin and water increased structuring. Thus, results indicated that beyond particles characteristics, which have already been widely studied in literature, the knowledge of the chemical and rheological properties of dispersing matrix is of great importance on structural behavior of suspensions / Doutorado / Doutor em Engenharia de Alimentos

No description available.

Orientador: Adriana Zerlotti Mercadante / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-09-11T21:13:51Z (GMT). No. of bitstreams: 1 Zepka_LeilaQueiroz_D.pdf: 2815017 bytes, checksum: 8abad3ae0da0bc1b940e100078096285 (MD5) Previous issue date: 2009 / Resumo: contém no texto completo. / Abstract: contém no texto completo. / Doutorado / Doutor em Ciência de Alimentos

No description available.

Die Rasterionenleitfähigkeitsmikroskopie (scanning ion conductance microscopy, SICM) stellt eine kontaktfreie Methode zur Ermittlung sowohl der Topographie als auch lokalen Ionenleitfähigkeit einer Oberfläche dar. Besonders vorteilhaft ist die Vermeidung mechanischer Beeinflussung bei der Untersuchung flexibler Strukturen, z.B. Lipiddoppelschichten wie Zellen oder künstlich erzeugter Lipidmembranen. Porenüberspannende Membranen (pore-suspending membranes, PSMs) verbinden als ein Beispiel für Modellsysteme eine hohe Stabilität mit lateraler Mobilität und dem Vorhandensein wässriger Kompartimente ober- und unterhalb der Doppelschicht, wie sie auch in der Natur gefunden werden. Ein wichtiges Forschungsgebiet stellt die Untersuchung der Wechselwirkung von Peptiden, besonders zellpenetrierenden Peptiden (cell penetrating peptides, CPPs), mit Lipiden und anderen Membranbestandteilen dar. Häufig untersuchte Beispiele sind Melittin, Hauptbestandteil des Giftes der Honigbiene Apis mellifera, sowie Penetratin, dritte Helix der Antennapedia Homöodomäne von Drosophila melanogaster. Generalisierte Protokolle zur Herstellung lösungsmittelfreier PSMs werden vorgestellt. Riesige unilamellare Vesikel (giant unilamellar vesicles, GUVs) unterschiedlicher Lipidzusammensetzung wurden hierzu auf porösem Siliziumnitrid (Si3N4), welches mit Cholesterylpolyethylenoxythiol (CPEO3, hydrophob) bzw. Mercaptoethanol (ME, hydrophil) funktionalisiert worden war, gespreitet. Verwendet wurden GUVs aus reinen Phosphatidylcholin (PC)-Lipiden sowie aus Mischungen von PC-Lipiden mit Cholesterol und PC-Lipiden mit Phosphatidylserin (PS)-Lipiden. Der Erfolg des Spreitvorgangs wurde durch Abbilden mittels konfokaler Rasterlasermikroskopie (confocal laser scanning microscopy, CLSM) und SICM verifiziert. Der Hauptteil dieser Arbeit behandelte die Entwicklung und Anwendung CLSM- und SICM-basierter CPP-Titrationsassays zur Aufklärung des Einflusses der Substratfunktionalisierung und der Lipidzusammensetzung der Membranen auf die Wechselwirkung zwischen Melittin bzw. Penetratin und den Lipiddoppelschichten. CLSM-Experimente wurden mit Melittin auf allen zur Verfügung stehenden PSMs sowohl auf hydrophob als auch hydrophil funktiona-lisierten Substraten durchgeführt, während Penetratin auf den drei unterschiedlichen PSMs auf hydrophil funktionalisierten Substraten verwendet wurde. Ein Reißen der Membranen wurde im Fall hydrophil funktionalisierter Substrate für beide Peptide im Bereich von 1–3 µM beobachtet. Bei hydrophob funktionalisierten Substraten induzierte eine dreifach geringere Melittinkonzentration die Zerstörung der Membranen. Sowohl auf hydrophob als auch auf hydrophil funktionalisierten Substraten wurde bei einem Cholesterolanteil von 10% eine Erhöhung der zum Reißen notwendigen Melittinkonzentratin erhalten, während bei 20% PS-Anteil eine Verschiebung zu geringeren Konzentrationen evident wurde. SICM-Experimente wurden mit Melittin auf PC/Cholesterol-PSMs auf hydrophob und hydrophil funktionalisierten Substraten und mit reinen PC-PSMs auf hydrophil funktionalisierten Membranen durchgeführt. Es wurden keine signifikanten Konzentrationsunterschiede beobachtet; die gefundenen Konzentrationsbereiche jedoch stimmten mit denen der CLSM-Experimente überein. Darüberhinaus wurde vor dem Reißen der Membranen ein Ansteigen der Porentiefe gefunden, das mit einer erhöhten Membranpermeabilität korrespondiert.

Master of Science / Food Science - Animal Sciences and Industry / J. Scott Smith / Heterocyclic amines (HCAs) are a class of mutagenic and carcinogenic chemical compounds formed on the outside of meat and fish when cooked at high temperatures. 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is the most abundantly formed HCA. HCAs have been found to cause cancer in mice and rats; PhIP specifically has been found to cause breast, rectal, prostate, and colon cancers. Model systems are often used to replicate the HCA chemical reactions in meat products without causing the many side reactions when meat is cooked at high temperatures. Model systems are also a useful way to study the effects of different variables and compounds on the formation of HCAs without using meat. A model system using amounts of 0.2 mmol glucose, 0.4 mmol creatinine, and 0.4 mmol phenylalanine in 10:90 water/diethylene glycol (v/v) was used to study the formation of PhIP. Differing levels of black pepper oil, black pepper extract, and rosemary extract (36, 71, 142, 285, 550 μL), synthetic antioxidants BHT and TBHQ (0.05 mmol, 0.1 mmol, 0.2 mmol, 0.4 mmol), and piperine (4.02 mg, 8.04 mg, 16.14 mg, 31.14 mg) were added to the model system to study their effect on PhIP formation. PhIP formation with added BHT (0.2 and 0.4 mmol) and TBHQ (0.4 mmol) were not significantly different from the control. All other added compounds decreased PhIP formation significantly from the control at p < 0.05. Solid phase micro extraction (SPME) headspace analysis was conducted on ground black pepper, black pepper oil, and black pepper extract to determine possible components responsible for PhIP inhibition. Six volatile compounds were found in common between ground black pepper, black pepper oil, and black pepper extract: 1R-α-pinene, 3-carene, caryophyllene, α-caryophyllene, cyclohexene, and D-limonene. D-limonene and caryophyllene had the largest peak areas, suggesting those compounds may play a part in PhIP inhibition in model systems.

Orientador: Neura Bragagnolo / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-17T18:17:21Z (GMT). No. of bitstreams: 1 Nogueira_GislaineChrystina_D.pdf: 842615 bytes, checksum: dfc01b70cfa919dc9a7f7698e623e3ea (MD5) Previous issue date: 2011 / Resumo: Uma das rotas de degradação do colesterol, quando submetido ao aquecimento em presença de oxigênio, é a formação de óxidos de colesterol (COP). Estes compostos têm sido associados a uma série de efeitos biológicos e doenças degenerativas e sua formação pode ser influenciada pela presença de outras substâncias, como antioxidantes e ácidos graxos. A influência dos antioxidantes está associada ao seu mecanismo de ação, que podem atuar nas fases de iniciação ou de propagação da oxidação lipídica. Já a influência dos ácidos graxos está associada com o grau de insaturação na molécula, em função da formação de radicais livres. Com o objetivo de verificar o comportamento do colesterol durante o aquecimento frente a influencia dessas substâncias, sistemas modelo contendo colesterol e ß-caroteno, TBHQ, vitamina E, ácido esteárico (saturado) ou ácido linolênico (com 3 insaturações), foram estudados. Os sistemas foram submetidos a 140, 180 e 220 °C, na presença de oxigênio, até que pelo menos 75% do colesterol fossem degradados. Os teores de colesterol, 7- hidroperoxicolesterol e COP foram avaliados ao longo do período de aquecimento. A degradação do colesterol foi observada em todas as temperaturas investigadas, ocorrendo em velocidade diretamente proporcional ao aumento da temperatura, totalizando aproximadamente 8 minutos a 220 °C, 20 minutos a 180 °C e 1 hora a 140 °C. De acordo com os resultados obtidos, a formação de COP atinge um máximo e a seguir sofre degradação, exceto na temperatura de 140 °C, em que o máximo da formação de COP só foi observado nos sistemas adicionados de ácidos graxos. A presença de ácidos graxos acelerou a formação de COP, entretanto, os sistemas contendo ácido linolênico apresentaram menores teores totais em todas as temperaturas estudadas e os sistemas contendo ácido esteárico apresentaram menores teores apenas a 220 °C, em comparação ao colesterol puro. Com exceção do TBHQ, a 140 °C, todos os antioxidantes estudados reduziram e/ou retardaram a formação de COP durante o aquecimento / Abstract: One of the pathways of cholesterol degradation, when it is submitted to heat, is the production of cholesterol oxidation products (COP). Those compounds have been associated to several biological effects and degenerative diseases and their formation can be influenced by other substances, such as antioxidants and fatty acids. The influence of the antioxidants is related to their mechanisms, which can act in the initiation and or propagation phases of the lipid oxidation. The influence of the fatty acids is associated to their degree of unsaturation, because of the free radical formation. In order to verify the cholesterol behavior during heating with the influence of those substances, model systems containing cholesterol and ß-carotene, TBHQ, vitamin E, stearic acid or linolenic acid, were studied. The model systems were heated at 140, 180 and 220 °C until at least 75% of cholesterol was degraded. The amount of cholesterol, COP and 7-hydroperoxycholesterol ¿ an intermediate compound of COP formation ¿ was evaluated during heating process. Cholesterol degradation was observed at all studied temperatures, occurring at a velocity directly proportional to the temperature increase, totalizing approximately 8 min at 220 °C, 20 min at 180 °C and 1 h at 140 °C. According to the obtained results, COP are formed reaching a maximum content and after that are degraded, but at 140 °C, the maximum was reached only at the systems containing fatty acids. The presence of fatty acids accelerated COP formation; however, the systems containing linolenic acid showed lower total content at all studied temperatures, on the other hand, the systems containing stearic acid showed lower content only at 220 °C, when compared to the pure cholesterol system. Except TBHQ, at 140 °C, all antioxidants were able to reduce and/or delay the maximum amount of total COP during heating / Doutorado / Doutor em Ciência de Alimentos

Concentration of Ce3+ is one of the most important parameters that influence the reactivity of ceria based catalyst. In this work we examine different experimental approaches for controlling Ce3+ concentration in cerium oxide model catalyst systems such as: i) influencing the stoichiometry of ceria, ii) introducing high valence doping agent, and iii) growing ultra thin ceria films with a strong metal substrate interaction. Structure, morphology and chemical state of prepared reduced ceria based systems were examined by means of surface science techniques: scanning tunneling microscopy, low-energy electron diffraction and X-ray photoelectron spectroscopy. In the present work an original method of ceria film reduction was introduced that allows stepwise control on stoichiometry and degree of film reduction (i). Further we introduce preparation procedures for well-ordered tungsten doped ceria model system (ii) and for the high quality 2D ultrathin ceria system on Cu (1 1 1) (iii). Preparation methods and model systems introduced in this work incorporate different physicochemical principles of Ce3+ induction and provide a variety of model systems useful for examining different effects that diversely prepared Ce3+ ions have on the activity of the catalyst.

<p>  </p> <p>Zebrafish (<em>Danio rerio</em>) is a well-established model to study neutrophil biology. However, a lack of standard tissue-specific knockdown or knockout technique in the zebrafish field has limited the power of this model organism when studying developmental essential genes, such as those related to mitochondrial function. We have developed a robust and flexible neutrophil-restricted knockout in zebrafish based on CRISPR/Cas9 system, with which we gained insights into the role of Rac2 in regulating the actin cytoskeleton and the subcellular location of Rac activation in zebrafish neutrophils.</p> <p>Previous study in our lab using another neutrophil-specific knockout system addressed multiple mitochondrial proteins regulate neutrophil motility in zebrafish. Interestingly, we observed <em>Mfn2</em>-deficient neutrophils trapped in the vasculature in zebrafish embryos. Here we further characterized the function of MFN2 in regulating cell migration with neutrophil-like HL-60 cells and mice embryonic fibroblasts (MEFs). We found significant changes in actin organization in both <em>MFN2</em>-deficient neutrophil-like cells and MEFs and mechanistically, disrupted mitochondria-ER interaction, increased intracellular Ca2+ levels. We also investigated the cytoskeleton proteins and observed hyperactivation of RhoA and Myosin light chain kinase, along with accumulation of phosphorylated myosin light chain at the cell boundary in <em>MFN2</em>-deficient MEFs. These altered MFN2-Ca2+-RhoA/MLCK-myosin signaling finally affects the peripheral actin bundle architecture and forms the “Peripheral Actin Myosin Belt (PAMB)” structure. The formation of PAMB hampered cell adhesive migration in <em>Mfn2</em>-null MEFs. </p> <p>Altogether, our research gained new insights into the essential role of MFN2 in cytoskeleton regulation and the underlying molecular mechanisms, which may provide a new direction to understand the relevance of this gene in immune cell dysfunction and other MFN2-associated diseases.</p>