The production, purification and characterisation of two novel alpha-D-mannosidases from Aspergillus phoenicis

1,6-alpha-D-Mannosidase from Aspergillits phoenicis was purified by anion-exchange chromatography, chromatofocussing and size-exclusion chromatography. The apparent molecular weight was 74 kDa by SDS-PAGE and 81 kDa by native-PAGE. The isoelectric point was 4.6. 1,6-alpha-D-Mannosidase had a temperature optimum of 60 degrees C, a pH optimum of 4.0-4.5. a K-m of 14 mM with alpha-D-Manp-(1 -> 6)-D-Manp as substrate. It was strongly inhibited by Mn2+ and did not need Ca2+ or any other metal cofactor of those tested. The enzyme cleaves specifically (1 -> 6)-linked mannobiose and has no activity towards any other linkages, p-nitrophenyl-alpha-D-mannopyranoside or baker's yeast mannan. 1,3(1,6)-alpha-D-Mannosidase from A. phoenicis was purified by anion-exchange chromatography, chromatofocus sing and size-exclusion chromatography. The apparent molecular weight was 97 kDa by SDS-PAGE and 110 kDa by native-PAGE. The 1,3(1,6)-alpha-D-mannosidase enzyme existed as two charge isomers or isoforms. The isoelectric points of these were 4.3 and 4.8 by isoelectric focussing. It cleaves alpha-D-Manp-(1 -> 3)-D-Manp 10 times faster than alpha-D-Manp-(1 -> 6)-D-Manp, has very low activity towards p-nitrophenyl-alpha-D-mannopyranoside and baker's yeast mannan, and no activity towards alpha-D-Manp-(1 -> 2)-D-Manp. The activity towards (1 -> 3)-linked mannobiose is strongly activated by 1 mM Ca2+ and inhibited by 10 mM EDTA, while (1 -> 6)-activity is unaffected, indicating that the two activities may be associated with different polypeptides. It is also possible that one polypeptide may have two active sites catalysing distinct activities. (c) 2005 Elsevier Ltd. All rights reserved.

Regioselective synthesis of mannobiose and mannotriose by reverse hydrolysis using a novel 1,6-alpha-D-mannosidase from Aspergillus phoenicis

A novel 1,6-alpha-D-mannosidase was produced by Aspergillus phoenicis grown on a commercial manno-oligosaccharide preparation in liquid culture. The enzyme hydrolysed only alpha-D-Manp-(1 --> 6)-D-Manp and did not act on alpha-D-Manp-(1 --> 2)-D-Manp, or alpha-D-Manp-(1 --> 3)-D-Manp. The 1,6-alpha-D-mannosidase was used for synthesis of manno-oligosaccharides by reverse hydrolysis reaction. The highest yields, expressed as percentages (w/w) of total sugar, were similar to21% mannobiose and similar to5% mannotriose, and they were obtained with 45% (w/w) initial mannose concentration at pH 4.5 after 12 days incubation at 55 degreesC. The disaccharide and trisaccharide products were separated and their structures determined by methylation analysis. Only 1-6 linkages were found in both of them. (C) 2003 Elsevier B.V. All rights reserved.

Singularities of optimal control problems on some 6-D lie groups

This paper considers the motion planning problem for oriented vehicles travelling at unit speed in a 3-D space. A Lie group formulation arises naturally and the vehicles are modeled as kinematic control systems with drift defined on the orthonormal frame bundles of particular Riemannian manifolds, specifically, the 3-D space forms Euclidean space E-3, the sphere S-3, and the hyperboloid H'. The corresponding frame bundles are equal to the Euclidean group of motions SE(3), the rotation group SO(4), and the Lorentz group SO (1, 3). The maximum principle of optimal control shifts the emphasis for these systems to the associated Hamiltonian formalism. For an integrable case, the extremal curves are explicitly expressed in terms of elliptic functions. In this paper, a study at the singularities of the extremal curves are given, which correspond to critical points of these elliptic functions. The extremal curves are characterized as the intersections of invariant surfaces and are illustrated graphically at the singular points. It. is then shown that the projections, of the extremals onto the base space, called elastica, at these singular points, are curves of constant curvature and torsion, which in turn implies that the oriented vehicles trace helices.

Object-based three-dimensional X-ray imaging

A form of three-dimensional X-ray imaging, called Object 3-D, is introduced, where the relevant subject material is represented as discrete ‘objects’. The surface of each such object is derived accurately from the projections of its outline, and of its other discontinuities, in about ten conventional X-ray views, distributed in solid angle. This technique is suitable for many applications, and permits dramatic savings in radiation exposure and in data acquisition and manipulation. It is well matched to user-friendly interactive displays.

Är teknikämnet efter över 20 år med egen kursplanpå väg att etablera sig i årskurs 1-3? : Åtta lärares beskrivning av sin planering och undervisning iteknik

Studien har haft som syfte att undersöka hur lärare i årskurs 1-3 planerar och genomförundervisning i teknik. Frågeställningarna som tagits fram för att för att få svar på studiens syfteär: hur beskriver lärarna att de planerar för teknikämnet, allmänt och i relation till det centralainnehållet för årskurs 1-3?, hur beskriver lärarna att de undervisar relaterat till det centralainnehållet i teknik för årskurs 1-3? samt vilka undervisningsmetoder används, enligt lärarna?Dessa frågor har det sökts svar på genom kvalitativa intervjuer med åtta lärare i årskurs 1-3.I resultatet framkommer att teknik är ett ämne som inte fullt ut har fått fäste i årskurs 1-3, då detvisar sig att flera lärare i studien saknar förtrogenheten med ämnets kursplan och planering ochundervisning beskrivs till stor del bedrivas tillsammans med No-ämnena. Teknikämnet har haften egen kursplan i tjugo år. Trots detta saknas på många skolor lärare med utbildning i tekniksamt material. Det framgår dock också i resultatet att några av de intervjuade lärarna ärintresserade och väl insatt i ämnets kursplan, även material i form av Skellefte-tekniken och NTAanvänds av flertalet av lärarna i studien. Teknikämnet behöver stärkas ytterligare som eget ämneoch i högre utsträckning undervisas separat för att ge eleverna en god uppfattning av vad som är teknik

Dynamics of glyphosate-induced conformational changes of Mycobacterium tuberculosis 5-enolpyruvylshikimate-3-phosphate synthase (EC 2.5.1.19) determined by hydrogen-deuterium exchange and electrospray mass spectrometry

The enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) catalyzes the reaction between shikimate 3-phosphate and phosphoenolpyruvate to form 5-enolpyruvylshikimate 3-phosphate, an intermediate in the shikimate pathway, which leads to the biosynthesis of aromatic amino acids. EPSPS exists in an open conformation in the absence of substrates and/or inhibitors and in a closed conformation when bound to the substrate and/or inhibitor. In the present report, the H/D exchange properties of EPSPS from Mycobacterium tuberculosis (Mt) were investigated for both enzyme conformations using ESI mass spectrometry and circular dichroism (CD). When the conformational changes identified by H/D exchanges were mapped on the 3-D structure, it was observed that the apoenzyme underwent extensive conformational changes due to glyphosate complexation, characterized by an increase in the content of alpha-helices from 40% to 57%, while the beta-sheet content decreased from 30% to 23%. These results indicate that the enzyme underwent a series of rearrangements of its secondary structure that were accompanied by a large decrease in solvent access to many different regions of the protein. This was attributed to the compaction of 71% of alpha-helices and 57% of beta-sheets as a consequence of glyphosate binding to the enzyme. Apparently, MtEPSPS undergoes a series of inhibitor-induced conformational changes, which seem to have caused synergistic effects in preventing solvent access to the core of molecule, especially in the cleft region. This may be part of the mechanism of inhibition of the enzyme, which is required to prevent the hydration of the substrate binding site and also to induce the cleft closure to avoid entrance of the substrates.

Bacterial cellulose/poly(3-hydroxybutyrate) composite membranes

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Structural characterization of the cell wall D-glucans isolated from the mycelium of Botryosphaeria rhodina MAMB-05

Three D-glucans were isolated from the mycelium of the fungus Botryosphaeria rhodina MAMB-05 by sequential extraction with hot-water and hot aqueous KOH (2% w/v) followed by ethanol precipitation. Following their purification by gel permeation chrornatography on Sepharose CL-4B, the structural characteristics of the D-glucans were determined by FT-IR and C-13 NMR spectroscopy and, after methylation, by GC-MS. The hot-water extract produced a fraction designated Q(1A) that was a beta-(1 -> 6)-D-glucan with the following structure:[GRAPHICS]The alkaline extract, when subjected to repeated freeze-thawing, yielded two fractions: KIP (insoluble) that comprised a beta-(1 -> 3)-D-glucan with beta-D-glucose branches at C-6 with the structure:[GRAPHICS]and K1SA (soluble) consisting of a backbone chain of alpha-(1 -> 4)-linked D-glucopyranosyl residues substituted at O-6 with alpha-D-glucopyranosyl residues:[GRAPHICS](c) 2008 Elsevier Ltd. All rights reserved.

Migração (2,5-D) com amplitudes verdadeiras em meios com gradiente constante de velocidade

A migração com amplitudes verdadeiras de dados de reflexão sísmica, em profundidade ou em tempo, possibilita que seja obtida uma medida dos coeficientes de reflexão dos chamados eventos de reflexão primária. Estes eventos são constituídos, por exemplo, pelas reflexões de ondas longitudinais P-P em refletores de curvaturas arbitrárias e suaves. Um dos métodos mais conhecido é o chamado migração de Kirchhoff, através do qual a imagem sísmica é produzida pela integração do campo de ondas sísmicas, utilizando-se superfícies de difrações, denominadas de Superfícies de Huygens. A fim de se obter uma estimativa dos coeficientes de reflexão durante a migração, isto é a correção do efeito do espalhamento geométrico, utiliza-se uma função peso no operador integral de migração. A obtenção desta função peso é feita pela solução assintótica da integral em pontos estacionários. Tanto no cálculo dos tempos de trânsito como na determinação da função peso, necessita-se do traçamento de raios, o que torna a migração em situações de forte heterogeneidade da propriedade física um processo com alto custo computacional. Neste trabalho é apresentado um algoritmo de migração em profundidade com amplitudes verdadeiras, para o caso em que se tem uma fonte sísmica pontual, sendo o modelo de velocidades em subsuperfície representado por uma função que varia em duas dimensões, e constante na terceira dimensão. Esta situação, conhecida como modelo dois-e-meio dimensional (2,5-D), possui características típicas de muitas situações de interesse na exploração do petróleo, como é o caso da aquisição de dados sísmicos 2-D com receptores ao longo de uma linha sísmica e fonte sísmica 3-D. Em particular, é dada ênfase ao caso em que a velocidade de propagação da onda sísmica varia linearmente com a profundidade. Outro tópico de grande importância abordado nesse trabalho diz respeito ao método de inversão sísmica denominado empilhamento duplo de difrações. Através do quociente de dois empilhamentos com pesos apropriados, pode-se determinar propriedades físicas e parâmetros geométricos relacionados com a trajetória do raio refletido, os quais podem ser utilizados a posteriori no processamento dos dados sísmicos, visando por exemplo, a análise de amplitudes.

Modelagem de eletrorresistividade 2-D a partir do potencial elétrico secundário

Um dos métodos clássicos da geofísica de exploração é o Método de Eletrorresistividade, estabelecido há um século pelos irmãos Schlumberger e desde então amplamente empregado em prospecção mineral, estudos ambientais e hidrogeologia e em pesquisa de fontes geotermais. Conceitualmente o método consiste de injeção de corrente elétrica na subsuperfície e de medida de diferença de potencial elétrico, resultante da interação da corrente com o meio. As localizações dessas fontes e receptores são determinadas pelo arranjo escolhido para o levantamento. Após o processamento, obtém-se pseudo-seções de resistividade aparente que indicam a distribuição de condutividade em subsuperfície. Devido à simplicidade dos fundamentos físicos de sua formulação, o método apresenta fácil implementação computacional quando comparado aos métodos eletromagnéticos de fonte controlada. Na literatura há inúmeros trabalhos de modelagem computacional, onde se calcula a resposta para problemas 2-D e 3-D. Nestes trabalhos, as pseudo-seções são obtidas a partir do cálculo do potencial elétrico total. Neste trabalho, apresentaremos a resposta da modelagem de eletroresistividade 2-D com o arranjo dipolo-dipolo, obtida a partir do potencial elétrico secundário. A solução é calculada através do método de elementos finitos usando malhas não estruturadas. Para efeito de validação, os resultados são comparados com a resposta 2-D obtida a partir dos potencias totais.

Production of β-(1,3)-glucanases by Trichoderma harzianum Rifai: Optimization and Application to Produce Gluco-oligosaccharides from Paramylon and Pustulan

β-(1→3)-Glucanases were produced by Trichoderma harzianum Rifai PAMB-86 cultivated on botryosphaeran in a bench-fermenter and optimised by the response surface method. Maximal enzyme titres occurred at 5 days, initial pH 5.5 and aeration of 1.5vvm. β-(1→3)-The β-glucanolytic enzyme complex produced by T. harzianum Rifai PAMB- 86 was fractionated by gel filtration into 2 fractions (F-I, F-II), and employed to produce gluco-oligosaccharides from algal paramylon ((1→3)-β-D-glucan) and lichen pustulan ((1→6)-β-D-glucan). Both enzymes attacked paramylon to the extent of ~15-20% in 30 min releasing glucose and laminaribiose as major end-products, and laminarioligosaccharides of degree of polymerization (DP) ≥3. Only F-I degraded pustulan resulting in ~2% degradation at 30 min, with glucose, gentiobiose and gentio-oligosaccharides of DP ≥4 as major products. The difference in the nature of the hydrolysis products can be explained by the substrate specificities of each enzyme fraction, and the structural differences of the β-D-glucans attacked.

The 2:1 cycloadducts from [3+2] 1,3-dipolar cycloaddition of nitrile oxide and vinylacetic acid. Synthesis and liquid crystal behaviour

Four liquid crystals (LC) 3,7a-bis(4-alkyloxyphenyl)-7,7a-dihydro-6H-isoxazolo[2,3-d][1,2,4]oxadiazol-6-yl)acetic acid (7a-d) were synthesised and the mesomorphic behaviour reported. The LCs were characterised as 2: 1 bisadducts, which were obtained from a double [3+2] 1,3-dipolar cycloaddition. In the first step, the cycloaddition of 4-alkyloxyphenylnitrile oxide (4a-d) and vinylacetic acid (5) gave the initial unobserved 1:1 cycloadducts 2-[3-(4-alkyloxyphenyl)-4,5-dihydroisoxazol-5-yl]acetic acid (6a-d). In the second step, the addition of a second equivalent of 4 to 6 yielded the 2: 1 bisadducts 7a-d without any traces of 6. All compounds 7a-d were unstable during the transition from the mesophase to the isotropic state upon first heating as evidenced by the large peaks in the differential scanning calorimetry traces. Due to the chemical instability of the compounds upon heating, the transition temperature related to the smectic C to smectic A transitions was acquired by means of an image processing method. X-Ray diffraction experiments were also used to analyse the liquid-crystalline phases. A theoretical calculation was performed using density functional theory (DFT) methods at the PBE1PBE/6-311+G(2d,p) level (with solvent effect) in order to get information about the energetic profile of the 2: 1 cycloaddition. DFT studies revealed that the cycloaddition process is controlled by the HOMO(dipolarophile) - LUMO(1,3-dipole), and that the double [3+2] 1,3-dipolar cycloaddition reaction is quite possible.

Contributo dei sistemi di fusione d'immagine mediante ecografia combinata con TAC e o RMN nella diagnosi del piccolo epatocarcinoma (< 2 cm) in pazienti con epatopatia cronica

L’ecografia è la metodica diagnostica più utilizzata come screening e follow-up nei pazienti epatopatici con o senza lesioni focali e questo grazie alle sue peculiari caratteristiche, che sono date dall’essere real-time, maneggevole, priva di radiazioni ionizzanti e con bassi costi. Tuttavia tale metodica se confrontata con la TC o la RMN, può avere importanti limiti, quali l’impossibilità di visualizzare piccole lesioni localizzate in aree anatomicamente “difficili” o in pazienti obesi, che sono già state identificate con altre tecniche, come la TC o la RMN. Per superare queste limitazioni sono stati introdotti dei sistemi di “fusione d’immagine” che consentono di sincronizzare in tempo reale una metodica real time con bassa risoluzione spaziale come l’ecografia ed una statica ad alta risoluzione come la TC o la RMN. Ciò si ottiene creando attorno al paziente un piccolo campo elettromagnetico costituito da un generatore e da un rilevatore applicato al trasduttore ecografico ed introducendo in un computer abbinato all’ecografo il “volume rendering” dell’addome del paziente ottenuto mediante TC multistrato o RM. Il preciso “ appaiamento spaziale “ delle due metodiche si ottiene individuando in entrambe lo stesso piano assiale di riferimento e almeno 3-4 punti anatomici interni. Tale sistema di fusione d’immagine potrebbe essere molto utile in campo epatologico nella diagnostica non invasiva del piccolo epatocarcinoma, che secondo le ultime linee guida, nei noduli di dimensioni fra 1 e 2 cm, richiede una concordanza nel comportamento contrastografico della lesione in almeno due tecniche d’immagine. Lo scopo del nostro lavoro è stato pertanto quello di valutare, in pazienti epatopatici, il contributo che tale sistema può dare nell’identificazione e caratterizzazione di lesioni inferiori a 20 mm, che erano già state identificate alla TC o alla RMN come noduli sospetti per HCC, ma che non erano stati visualizzati in ecografia convenzionale. L’eventuale re-identificazione con l’ecografia convenzionale dei noduli sospetti per essere HCC, può permettere di evitare, alla luce dei criteri diagnostici non invasivi un’ ulteriore tecnica d’immagine ed eventualmente la biopsia. Pazienti e Metodi: 17 pazienti cirrotici (12 Maschi; 5 Femmine), con età media di 68.9 +/- 6.2 (SD) anni, in cui la TC e la RMN con mezzo di contrasto avevano identificato 20 nuove lesioni focali epatiche, inferiori a 20 mm (13,6 +/- 3,6 mm), sospette per essere epatocarcinomi (HCC), ma non identificate all’ecografia basale (eseguita in cieco rispetto alla TC o alla RMN) sono stati sottoposti ad ecografia senza e con mezzo di contrasto, focalizzata su una zona bersaglio identificata tramite il sistema di fusione d’immagini, che visualizza simultaneamente le immagini della TC e della RMN ricostruite in modalità bidimensionale ( 2D), tridimensionale ( 3 D) e real-time. La diagnosi finale era stata stabilita attraverso la presenza di una concordanza diagnostica, secondo le linee guida internazionali o attraverso un follow-up nei casi di discordanza. Risultati: Una diagnosi non invasiva di HCC è stata raggiunta in 15/20 lesioni, inizialmente sospettate di essere HCC. Il sistema di fusione ha identificato e mostrato un comportamento contrastografico tipico in 12/15 noduli di HCC ( 80%) mentre 3/15 HCC (20%) non sono stati identificati con il sistema di fusione d’immagine. Le rimanenti 5/20 lesioni non sono state visualizzate attraverso i sistemi di fusione d’immagine ed infine giudicate come falsi positivi della TC e della RMN, poiché sono scomparse nei successivi mesi di follow-up e rispettivamente dopo tre, sei, nove, dodici e quindici mesi. Conclusioni: I nostri risultati preliminari mostrano che la combinazione del sistema di fusione dell’immagine associata all’ecografia senza e con mezzo di contrasto (CEUS), migliora il potenziale dell’ecografia nell’identificazione e caratterizzazione dell’HCC su fegato cirrotico, permettendo il raggiungimento di una diagnosi, secondo criteri non invasivi e slatentizzazndo casi di falsi positivi della TC e della RMN.

HCHO, H 2 O 2 und CH 3 OOH in der Troposphäre

In dieser Arbeit wurden im Rahmen der UTOPIHAN- und HOHPEX04-Projekte Peroxid- und Formaldehydmessungen in der Troposphäre durchgeführt und wissenschaftlich interpretiert. Die Messungen während UTOPIHAN fanden dabei an Bord eines für Forschungszwecke umgerüsteten Flugzeuges (Learjet 35A) im Wesentlichen in der freien, insbesondere in der oberen Troposphäre über Europa statt. Die Messungen während HOHPEX04 waren hingegen als Bodenmessungen an der sich abwechselnd in der bodennahen Grenzschicht und in von dieser Schicht entkoppelten Luftmassen liegenden Bergstation Hohenpeißenberg (bayerisches Voralpenland) konzipiert. Um eine quantitative Auswertbarkeit der Messungen sicherzustellen, wurden die verwendeten, auf chemischer Derivatisierung und fluorimetrischer Detektion basierenden Messgeräte AL 2001CA (Peroxide) und AL 4021 (Formaldehyd) (AEROLASER) genau charakterisiert. Dabei wurde speziell die bekannte Ozoninterferenz beider Geräte in einer großen Zahl von Laborexperimenten mit unterschiedlichen Randbedingungen bezüglich Wasserdampf- und Kohlenwasserstoffgehalt der Luft untersucht. Für beide Verbindungen wurden Höhen- sowie Breitenprofile erstellt und mit Ergebnissen eines 3D-Chemie-Transport-Modells (CTM) sowie früherer Studien verglichen. In einem weiteren Kapitel werden Ergebnisse einer quantitativen Studie zum Einfluss hochreichender Konvektion auf das HCHO-Budget in der mittleren und oberen Troposphäre präsentiert. Diese Studie kommt zu dem Schluss, dass der rasche Aufwärtstransport von Vorläufergasen von HCHO und HOx wie Methanol, Aceton und sogar gut löslicher Spurengase wie CH3OOH beziehungsweise H2O2 aus der Grenzschicht einen signifikanten, auf Grund der längeren Lebensdauer von NOx über mehrere Tage andauernden und damit großräumigen Einfluss auf die Budgets von HCHO, HOx und auch O3 in der oberen Troposphäre haben kann. Die Befunde der Studie legen desweiteren nahe, dass fehlerhafte Modellvorhersagen für die NO-Mischungsverhältnisse in der Tropopausenregion, die zum Beispiel mit Mängeln des Modells bezüglich der Höhe der Konvektion und des Stratosphären-Troposphären-Austauschs zu tun haben, hauptverantwortlich sind für gefundene Differenzen zwischen Messdaten und dem verwendeten 3D-Chemie-Transport-Modell. Um die Signifikanz der Aussagen zu erhöhen, wurde eine Sensitivitätsstudie durchgeführt, in der die Konzentration einiger chemischer Verbindungen sowie die Photolyseraten variiert wurden. Eine weitere Studie zum Einfluss verschiedener Parameter auf das CH3OOH/H2O2-Verhältnis kommt zu dem Schluss, dass dieses Verhältnis keinen idealen Indikator für Wolkenprozessierung von Luftmassen darstellt, während eine signifikant positive Abweichung vom H2O2/H2O-Verhältnis in der oberen Troposphäre ein guter Indikator für rasch aufwärts transportierte Luftmassen sein kann. Im Rahmen dieser Studie werden auch Höhen- und Breitenprofile des CH3OOH/H2O2-Verhältnisses diskutiert. In einer letzten Untersuchung zu HCHO-Messungen am Observatorium Hohenpeißenberg im Sommer 2004 werden für die in zwei Windrichtungssektoren eingeteilten Daten Korrelationen anderer Spurengase wie O3, PAN, CO, NOy und Isopren mit HCHO interpretiert. In diesem Zusammenhang wird auch versucht, den beobachteten Tagesgang von HCHO zu erklären.

EXPERIMENTAL AND COMPUTATIONAL STUDIES OF ELECTROMAGNETIC CLOAKING AT MICROWAVES

An invisibility cloak is a device that can hide the target by enclosing it from the incident radiation. This intriguing device has attracted a lot of attention since it was first implemented at a microwave frequency in 2006. However, the problems of existing cloak designs prevent them from being widely applied in practice. In this dissertation, we try to remove or alleviate the three constraints for practical applications imposed by loosy cloaking media, high implementation complexity, and small size of hidden objects compared to the incident wavelength. To facilitate cloaking design and experimental characterization, several devices and relevant techniques for measuring the complex permittivity of dielectric materials at microwave frequencies are developed. In particular, a unique parallel plate waveguide chamber has been set up to automatically map the electromagnetic (EM) field distribution for wave propagation through the resonator arrays and cloaking structures. The total scattering cross section of the cloaking structures was derived based on the measured scattering field by using this apparatus. To overcome the adverse effects of lossy cloaking media, microwave cloaks composed of identical dielectric resonators made of low loss ceramic materials are designed and implemented. The effective permeability dispersion was provided by tailoring dielectric resonator filling fractions. The cloak performances had been verified by full-wave simulation of true multi-resonator structures and experimental measurements of the fabricated prototypes. With the aim to reduce the implementation complexity caused by metamaterials employment for cloaking, we proposed to design 2-D cylindrical cloaks and 3-D spherical cloaks by using multi-layer ordinary dielectric material (εr>1) coating. Genetic algorithm was employed to optimize the dielectric profiles of the cloaking shells to provide the minimum scattering cross sections of the cloaked targets. The designed cloaks can be easily scaled to various operating frequencies. The simulation results show that the multi-layer cylindrical cloak essentially outperforms the similarly sized metamaterials-based cloak designed by using the transformation optics-based reduced parameters. For the designed spherical cloak, the simulated scattering pattern shows that the total scattering cross section is greatly reduced. In addition, the scattering in specific directions could be significantly reduced. It is shown that the cloaking efficiency for larger targets could be improved by employing lossy materials in the shell. At last, we propose to hide a target inside a waveguide structure filled with only epsilon near zero materials, which are easy to implement in practice. The cloaking efficiency of this method, which was found to increase for large targets, has been confirmed both theoretically and by simulations.