Nutritional and anti-nutritional analyses of Azolla africana Desv. and Spirodela polyrrhiza L. Schleiden as feedstuffs for fish production

The chemical composition of Azolla africana and Spirodela polyrrhiza cultivated in earthen ponds were determined. Crude protein contents of the samples were 28.9~c0.6 and 25.6~c0.2% dry matter for A. africana and S. polyrrhiza respectively. Dry matter, crude fibre and lipid contents of A. africana were higher (P<0.05) than values obtained for S. polyrrhiza. Mineral analyses showed that S. polyrrhiza contained higher levels of Na, S, Ca, Mg and Fe than A. africana. Except for Ca content in S. polyrrhiza, heavy metals (Ni and Zn) accumulation in Azolla were very high. There were no wide differences in the individual amino acid indexes except for methionine. Some anti-nutritional factors were determined. Cyanide, tannin and phytin contents of fresh weed samples were higher than sun-dried samples. A. africana contained more cyanide and tannin than S. polyrrhiza both in fresh and sun-dried forms

Relation between TMAOase activity and content of formaldehyde in fillet minces and bellyflap minces from gadoid fishes

Minced fish is a significant component of a number of frozen fishery products like fish fingers, cakes and patties. Predominately minced fish is produced from gadoid species (Alaska pollack, cod, saithe, hake and others) possessing the enzyme trimethylamine oxide demethylase (TMAOase, E.C. 4.1.2.32) (Rehbein and Schreiber 1984). TMAOase catalyses the degradation of trimethylamine oxide (TMAO) to formaldehyde (FA) and dimethylamine (DMA), preferentially during frozen storage of products (Hultin 1992). In most gadoid species light muscle contains only low activity of TMAOase, the activity of red muscle and bellyflaps being somewhat higher. In contrast, the TMAOase activity in blood, kidney and other tissues, residues of which may contaminate minced fish flesh, may be higher for several orders of magnitude (Rehbein and Schreiber 1984).

Proteomic analysis of the Cdc48/Ubx network identifies a role for Ubx2 in the regulation of lipid biosynthesis

Cdc48/p97 is an essential, highly abundant hexameric member of the AAA (ATPase associated with various cellular activities) family. It has been linked to a variety of processes throughout the cell but it is best known for its role in the ubiquitin proteasome pathway. In this system it is believed that Cdc48 behaves as a segregase, transducing the chemical energy of ATP hydrolysis into mechanical force to separate ubiquitin-conjugated proteins from their tightly-bound partners.

Current models posit that Cdc48 is linked to its substrates through a variety of adaptor proteins, including a family of seven proteins (13 in humans) that contain a Cdc48-binding UBX domain. As such, due to the complexity of the network of adaptor proteins for which it serves as the hub, Cdc48/p97 has the potential to exert a profound influence on the ubiquitin proteasome pathway. However, the number of known substrates of Cdc48/p97 remains relatively small, and smaller still is the number of substrates that have been linked to a specific UBX domain protein. As such, the goal of this dissertation research has been to discover new substrates and better understand the functions of the Cdc48 network. With this objective in mind, we established a proteomic screen to assemble a catalog of candidate substrate/targets of the Ubx adaptor system.

Here we describe the implementation and optimization of a cutting-edge quantitative mass spectrometry method to measure relative changes in the Saccharomyces cerevisiae proteome. Utilizing this technology, and in order to better understand the breadth of function of Cdc48 and its adaptors, we then performed a global screen to identify accumulating ubiquitin conjugates in cdc48-3 and ubxΔ mutants. In this screen different ubx mutants exhibited reproducible patterns of conjugate accumulation that differed greatly from each other, pointing to various unexpected functional specializations of the individual Ubx proteins.

As validation of our mass spectrometry findings, we then examined in detail the endoplasmic-reticulum bound transcription factor Spt23, which we identified as a putative Ubx2 substrate. In these studies ubx2Δ cells were deficient in processing of Spt23 to its active p90 form, and in localizing p90 to the nucleus. Additionally, consistent with reduced processing of Spt23, ubx2Δ cells demonstrated a defect in expression of their target gene OLE1, a fatty acid desaturase. Overall, this work demonstrates the power of proteomics as a tool to identify new targets of various pathways and reveals Ubx2 as a key regulator lipid membrane biosynthesis.

Cooperative properties of lipid bilayers : collective director fluctuations and the effects of hydrophobic mismatch in protein/lipid membrane systems

Cooperative director fluctuations in lipid bilayers have been postulated for many years. ^2H-NMR T_1^(-1), T_(1P)^(-1) , and T_2^(-1); measurements have been used identify these motions and to determine the origin of increased slow bilayer motion upon addition of unlike lipids or proteins to a pure lipid bilayer.

The contribution of cooperative director fluctuations to NMR relaxation in lipid bilayers has been expressed mathematically using the approach of Doane et al.^1 and Pace and Chan.^2 The T_2^(-1)’s of pure dimyristoyllecithin (DML) bilayers deuterated at the 2, 9 and 10, and all positions on both lipid hydrocarbon chains have been measured. Several characteristics of these measurements indicate the presence of cooperative director fluctuations. First of all, T_2^(-1) exhibits a linear dependence on S2/CD. Secondly, T_2^(-1) varies across the ^2H-NMR powder pattern as sin^2 (2, β), where , β is the angle between the average bilayer director and the external magnetic field. Furthermore, these fluctuations are restricted near the lecithin head group suggesting that the head group does not participate in these motions but, rather, anchors the hydrocarbon chains in the bilayer.

T_2^(-1)has been measured for selectively deuterated liquid crystalline DML hilayers to which a host of other lipids and proteins have been added. The T_2^(-1) of the DML bilayer is found to increase drastically when chlorophyll a (chl a) and Gramicidin A' (GA') are added to the bilayer. Both these molecules interfere with the lecithin head group spacing in the bilayer. Molecules such as myristic acid, distearoyllecithin (DSL), phytol, and cholesterol, whose hydrocarbon regions are quite different from DML but which have small,neutral polar head groups, leave cooperative fluctuations in the DML bilayer unchanged.

The effect of chl a on cooperative fluctuations in the DML bilayer has been examined in detail using ^2H-NMR T_1^(-1), T_(1P)^(-1) , and T_2^(-1); measurements. Cooperative fluctuations have been modelled using the continuum theory of the nematic state of liquid crystals. Chl a is found to decrease both the correlation length and the elastic constants in the DML bilayer.

A mismatch between the hydrophobic length of a lipid bilayer and that of an added protein has also been found to change the cooperative properties of the lecithin bilayer. Hydrophobic mismatch has been studied in a series GA' / lecithin bilayers. The dependence of 2H-NMR order parameters and relaxation rates on GA' concentration has been measured in selectively deuterated DML, dipalmitoyllecithin (DPL), and DSL systems. Order parameters, cooperative lengths, and elastic constants of the DML bilayer are most disrupted by GA', while the DSL bilayer is the least perturbed by GA'. Thus, it is concluded that the hydrophobic length of GA' best matches that of the DSL bilayer. Preliminary Raman spectroscopy and Differential Scanning Calorimetry experiments of GA' /lecithin systems support this conclusion. Accommodation of hydrophobic mismatch is used to rationalize the absence of H_(II) phase formation in GA' /DML systems and the observation of H_(II) phase in GA' /DPL and GA' /DSL systems.

1. J. W. Doane and D. L. Johnson, Chem. Phy3. Lett., 6, 291-295 (1970). 2. R. J. Pace and S. I. Chan, J. Chem. Phy3., 16, 4217-4227 (1982).

Chemical studies of viral entry mechanisms: I. Hydrophobic protein-lipid interactions during sendai virus membrane fusion. II. Kinetics of bacteriophage λ DNA injection.

Viruses possess very specific methods of targeting and entering cells. These methods would be extremely useful if they could also be applied to drug delivery, but little is known about the molecular mechanisms of the viral entry process. In order to gain further insight into mechanisms of viral entry, chemical and spectroscopic studies in two systems were conducted, examining hydrophobic protein-lipid interactions during Sendai virus membrane fusion, and the kinetics of bacteriophage λ DNA injection.

Sendai virus glycoprotein interactions with target membranes during the early stages of fusion were examined using time-resolved hydrophobic photoaffinity labeling with the lipid-soluble carbene generator3-(trifluoromethyl)-3-(m-^(125 )I] iodophenyl)diazirine (TID). The probe was incorporated in target membranes prior to virus addition and photolysis. During Sendai virus fusion with liposomes composed of cardiolipin (CL) or phosphatidylserine (PS), the viral fusion (F) protein is preferentially labeled at early time points, supporting the hypothesis that hydrophobic interaction of the fusion peptide at the N-terminus of the F_1 subunit with the target membrane is an initiating event in fusion. Correlation of the hydrophobic interactions with independently monitored fusion kinetics further supports this conclusion. Separation of proteins after labeling shows that the F_1 subunit, containing the putative hydrophobic fusion sequence, is exclusively labeled, and that the F_2 subunit does not participate in fusion. Labeling shows temperature and pH dependence consistent with a need for protein conformational mobility and fusion at neutral pH. Higher amounts of labeling during fusion with CL vesicles than during virus-PS vesicle fusion reflects membrane packing regulation of peptide insertion into target membranes. Labeling of the viral hemagglutinin/neuraminidase (HN) at low pH indicates that HN-mediated fusion is triggered by hydrophobic interactions, after titration of acidic amino acids. HN labeling under nonfusogenic conditions reveals that viral binding may involve hydrophobic as well as electrostatic interactions. Controls for diffusional labeling exclude a major contribution from this source. Labeling during reconstituted Sendai virus envelope-liposome fusion shows that functional reconstitution involves protein retention of the ability to undergo hydrophobic interactions.

Examination of Sendai virus fusion with erythrocyte membranes indicates that hydrophobic interactions also trigger fusion between biological membranes, and that HN binding may involve hydrophobic interactions as well. Labeling of the erythrocyte membranes revealed close membrane association of spectrin, which may play a role in regulating membrane fusion. The data show that hydrophobic fusion protein interaction with both artificial and biological membranes is a triggering event in fusion. Correlation of these results with earlier studies of membrane hydration and fusion kinetics provides a more detailed view of the mechanism of fusion.

The kinetics of DNA injection by bacteriophage λ. into liposomes bearing reconstituted receptors were measured using fluorescence spectroscopy. LamB, the bacteriophage receptor, was extracted from bacteria and reconstituted into liposomes by detergent removal dialysis. The DNA binding fluorophore ethidium bromide was encapsulated in the liposomes during dialysis. Enhanced fluorescence of ethidium bromide upon binding to injected DNA was monitored, and showed that injection is a rapid, one-step process. The bimolecular rate law, determined by the method of initial rates, revealed that injection occurs several times faster than indicated by earlier studies employing indirect assays.

It is hoped that these studies will increase the understanding of the mechanisms of virus entry into cells, and to facilitate the development of virus-mimetic drug delivery strategies.

One-step digit-set-restricted modified signed-digit adder using an incoherent correlator based on a shared content-addressable memory

An efficient one-step digit-set-restricted modified signed-digit (MSD) adder based on symbolic substitution is presented. In this technique, carry propagation is avoided by introducing reference digits to restrict the intermediate carry and sum digits to {1,0} and {0,1}, respectively. The proposed technique requires significantly fewer minterms and simplifies system complexity compared to the reported one-step MSD addition techniques. An incoherent correlator based on an optoelectronic shared content-addressable memory processor is suggested to perform the addition operation. In this technique, only one set of minterms needs to be stored, independent of the operand length. (C) 2002 society or Photo-Optical Instrumentation Engineers.

Efficient two-step digit-set-restricted modified signed-digit algorithm based on optoelectronic shared content-addressable memory

A two-step digit-set-restricted modified signed-digit (MSD) adder based on symbolic substitution is presented. In the proposed addition algorithm, carry propagation is avoided by using reference digits to restrict the intermediate MSD carry and sum digits into {(1) over bar ,0} and {0, 1}, respectively. The algorithm requires only 12 minterms to generate the final results, and no complementarity operations for nonzero outputs are involved, which simplifies the system complexity significantly. An optoelectronic shared content-addressable memory based on an incoherent correlator is used for experimental demonstration. (c) 2005 Society of Photo-Optical Instrumentation Engineers.

Efficient two-step digit-set-restricted modified signed-digit algorithm based on optoelectronic shared content-addressable memory

A two-step digit-set-restricted modified signed-digit (MSD) adder based on symbolic substitution is presented. In the proposed addition algorithm, carry propagation is avoided by using reference digits to restrict the intermediate MSD carry and sum digits into {(1) over bar ,0} and {0, 1}, respectively. The algorithm requires only 12 minterms to generate the final results, and no complementarity operations for nonzero outputs are involved, which simplifies the system complexity significantly. An optoelectronic shared content-addressable memory based on an incoherent correlator is used for experimental demonstration. (c) 2005 Society of Photo-Optical Instrumentation Engineers.

The content of organic matter in some waters in the Moscow region [Translation from: C.R. Acad. Soc. URSS, 61, 293-296, 1948]

There is at the moment no direct method of determining the organic matter content of natural waters. In 1940/41 8 different water bodies in central Russia were studied and their organic matter identified. The author concludes that there is currently no easy method to determine organic matter in water. A number methods need to be applied.

Some data on the content of organic carbon in rivers in the winter months. [Translation from: Gidrochim. Mat. 23, 36-38, 1955]

Observations are reported on the content of organic matter in the Moscow region in 1941. Some data is given on alkalinity, oxygen content and colourisation of the rivers.

Improving Content Delivery Efficiency through Multi-Layer Mobile Edge Adaptation

This paper presents a novel architecture for optimizing the HTTP-based multimedia delivery in multi-user mobile networks. This proposal combines the usual client-driven dynamic adaptation scheme DASH-3GPP with network-assisted adaptation capabilities, in order to maximize the overall Quality of Experience. The foundation of this combined adaptation scheme is based on two state of the art technologies. On one hand, adaptive HTTP streaming with multi-layer encoding allows efficient media delivery and improves the experienced media quality in highly dynamic channels. Additionally, it enables the possibility to implement network-level adaptations for better coping with multi-user scenarios. On the other hand, mobile edge computing facilitates the deployment of mobile services close to the user. This approach brings new possibilities in modern and future mobile networks, such as close to zero delays and awareness of the radio status. The proposal in this paper introduces a novel element, denoted as Mobile Edge-DASH Adaptation Function, which combines all these advantages to support efficient media delivery in mobile multi-user scenarios. Furthermore, we evaluate the performance enhancements of this content- and user context-aware scheme through simulations of a mobile multimedia scenario.

Estudo da bioenergética mitocondrial de cardiomiócitos de camundongos obesos

A obesidade é uma doença crônica, resultante do excesso de gordura no organismo. O aumento da obesidade no mundo, tem se revelado como um dos fenômenos clínicos e epidemiológicos da atualidade. Estudos populacionais e em modelos animais demonstram que a origem da epidemia da obesidade está relacionada a fatores genéticos, modificações de hábitos nutricionais, redução da atividade física, e alterações nutricionais durante a lactação, desempenhando um papel relevante no desenvolvimento da obesidade, DM2 e cardiomiopatias. As mitocôndrias são os coordenadores centrais do metabolismo energético, assim, alterações funcionais e estruturais dessa organela têm sido associadas à desordens metabólicas. Elas exercem um papel na sobrevivência e função dos cardiomiócitos devido à alta demanda energética do miocárdio. Desta forma, disfunções mitocondriais estão relacionadas com disfunções no miocárdio e conseqüente progressão de cardiomiopatias. Neste estudo, avaliamos a bioenergética e a ultraestrutura de cardiomiócitos de camundongos obesos e controle hiperalimentados durante a lactação. O consumo de oxigênio das fibras cardíacas foi avaliado por respirometria de alta-resolução, utilizando um oxígrafo-2K-Oroboros. A ultraestrutura dos cardiomiócitos foi analisada por microscopia eletrônica de transmissão e o conteúdo das proteínas Carnitina palmitoil transferase 1 (CPT1), Proteína desacopladora 2 (UCP2) , Transportador de glicose 1 e 4 (GLUT1) e (GLUT4), Proteína Kinase ativada por AMP (AMPK) e Proteína kinase ativada por AMP fosforilada p(AMPK) por Western blotting (WB). Além disso, o peso dos animais, a gordura retroperitoneal, epididimal e a glicemia em jejum foram determinadas. Nossos resultados confirmaram que os animais do grupo hiperalimentados (GH), aos 90 dias de vida, apresentaram aumento da massa corporal, de gordura epididimária e retroperitoneal comparado ao grupo controle (GC). As taxas respiratórias foram semelhantes nos dois grupos quando foram utilizados os substratos dos complexos I e II. Entretanto, quando o ácido graxo palmitoil-L-carnitina foi utilizado, a taxa respiratória máxima do GH foi significativamente menor. A análise ultraestrutural dos cardiomiócitos do GH demonstrou intenso dano na matriz mitocondrial e maior presença de gotículas de lipídios, caracterizando deposição ectópica. Os resultados do WB mostraram aumento significativo do conteúdo de CPT1 e UCP2 no GH comparado ao GC. Não foram encontradas diferenças significativas no conteúdo de GLUT1 entre os grupos, entretanto, observamos maior conteúdo do GLUT4 no GH. Além disso, encontramos maior conteúdo de AMPK no GH, ao passo que o conteúdo de pAMPK foi semelhante entre os grupos. Entretanto, a razão pAMPK/AMPK é significativamente menor no GH. Esses resultados sugerem que a hiperalimentação durante a lactação leva a obesidade na vida adulta com alterações na bioenergética e ultraestrutura dos cardiomiócitos.

Combination of web usage, content and structure information for diverse web mining applications in the tourism context and the context of users with disabilities

188 p.