Wyniki wyszukiwania - Biblioteka Nauki

1

Expression of human serum albumin - L7-L12 [Brucella abortus ribosomal protein] fusion protein in Saccharomyces cerevisiae

100%

Pakzad I. , Rezaee A. , Emaneini M. , Hosseini A. Z. , Tabbaraee B. , Taherikalani M.

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nr 2

99-104

EN

Brucella abortus is a facultative intracellular gram-negative bacterial pathogen that causes abortion in pregnant cattle and undulant fever in humans. The immunogenic B. abortus ribosomal protein L7/L12 is a promising candidate antigen for the development of subunit vaccines against brucellosis. It has already been expressed in several bacteria and has been used as DNA vaccine. In order to construct yeast expressing vector for the tHSA-L7/L12 fusion protein, the 171112 ribosomal gene was amplified by PCR. The expression plasmid pYtHSA-L7/Ll 2 was constructed by inserting the L7/L12 gene into the pYHSA5 shuttle vector (containing inulinase signal sequence, HSA gene and Gal 10 promoter). The recombinant vector was transformed into S. cerevisiae and was then induced by galactose. The secreted recombinant fusion protein was detected in supernatant by SDS-PAGE and confirmed by western blot analysis using anti-HSA and anti-L7/L 12 antibodies. Fusion protein was purified by affinity chromatography and its amount was approximately 500 μg/liter.

2

A comparative study on translational diff usion coeffi cient for some mammalian serum albumins

100%

Monkos K.

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tom 64

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nr 5-6

43-53

EN

BACKGROUND The aim of the present paper is investigation of the volume fraction dependence of the translational diffusion coefficient for some mammalian serum albumins in aqueous solutions. MATERIAL AND METHODS The viscosity of bovine, equine, ovine and rabbit serum albumin aqueous solutions was measured at temperatures ranging from 5oC to 45oC and in a wide range of concentrations. The measurements were performed with an Ubbelohde-type capillary microviscometer. RESULTS Translational diffusion coefficient at infinitely dilute solutions Do(T) can be calculated from generalized Stokes-Einstein equation if the hydrodynamic radius of albumin is known. It gives Do(T) in the range from 3.5×10-11 m2 /s (at 5oC) to 10.2 10-11 m2 /s (at 45oC) for bovine serum albumin, from 3.59×10-11 m2 /s (at 5oC) to 10.4 10-11 m2 /s (at 45oC) for equine serum albumin, from 3.42×10-11 m2 /s (at 5oC) to 9.92×10-11 m2 /s (at 45oC) for ovine serum albumin, and from 3.36×10-11 m2 /s (at 5oC) to 9.74×10-11 m2 /s (at 45oC) for rabbit serum albumin. Translational diffusion coefficient for higher concentrations D(T,φ) can be obtained from the relation: D(T,φ) = Do(T)Ηo(T)/Η(T,φ), where φ denotes volume fraction and Ηo(T) and Η(T,φ) are viscosities of water and solution, respectively, at temperature T. CONCLUSIONS The obtained results show that the translational diffusion coefficient decreases linearly with increasing volume fraction, when φ does not exceed the value of about 0.1. The dependence of the translational diffusion coefficient on volume fraction in a broader range of φ, i.e. from dilute to concentrated solutions, is nonlinear and can be described by a stretched exponential function.

PL

WSTĘP Celem niniejszej pracy jest zbadanie zależności współczynnika dyfuzji translacyjnej od ułamka objętościowego dla albumin surowicy kilku ssaków w roztworach wodnych. MATERIAŁ I METODY Lepkość wodnych roztworów albuminy surowicy wołowej, końskiej, owczej i króliczej zmierzono w zakresie temperatur od 5oC do 45oC i w szerokim zakresie stężeń. Pomiary wykonano przy pomocy kapilarnego mikrowiskozymetru typu Ubbelohde’a. WYNIKI Współczynnik dyfuzji translacyjnej dla roztworów rozcieńczonych Do(T) można obliczyć z uogólnionego równania Stokesa-Einsteina, jeżeli znany jest promień hydrodynamiczny albuminy. Daje to Do(T) w zakresie od 3.5×10-11 m2 /s (w 5oC) do 10.2×10-11 m2 /s (w 45oC) dla albuminy surowicy wołowej, od 3.59×10-11 m2 /s (w 5oC) do 10.4×10-11 m2 /s (w 45oC) dla albuminy surowicy końskiej, od 3.42×10-11 m2 /s (w 5oC) do 9.92×10-11 m2 /s (w 45oC) dla albuminy surowicy owczej i od 3.36×10-11 m2 /s (w 5oC) do 9.74×10-11 m2 /s (w 45oC) dla albuminy surowicy króliczej. Współczynnik dyfuzji translacyjnej dla wyższych stężeń D(T,φ) można otrzymać z relacji: D(T,φ) = Do(T)Ηo(T)/Η(T,φ), w której φ oznacza ułamek objętościowy a Ηo(T) i Η(T,φ) są, odpowiednio, lepkością wody i roztworu w temperaturze T. WNIOSKI Otrzymane wyniki pokazują, że współczynnik dyfuzji translacyjnej maleje liniowo wraz ze wzrostem ułamka objętościowego, jeżeli φ nie przekracza wartości około 0.1. Zależność współczynnika dyfuzji translacyjnej od ułamka objętościowego w szerszym zakresie φ , tzn. dla roztworów od rozcieńczonych do stężonych, jest nieliniowa i można ją opisać rozciągniętą funkcją wykładniczą.

3

Interaction of anthracene and its oxidative derivatives with human serum albumin

100%

Skupinska K. , Zylm M. , Misiewicz I. , Kasprzycka-Guttman T.

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tom 53

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nr 1

EN

Binding affinities of ten polycyclic aromatic hydrocarbons to albumin were determined: anthracene, its eight oxy-derivatives: anthraquinone, 9-anthracenemethanol, 9-anthraldehyde, 9-anthracenecarboxylic acid, 1,4-dihydroxyanthraquinone, 1,5-dihydroxyanthraquinone, 1,8-dihydroxyanthraquinone, 2,6-dihydroxyanthraquinone and benzo[a]pyrene. The quenching of albumin fluorescence was used to measure the PAH — protein interaction. The theoretical curve of calculated fluorescence was fitted to experimental data after necessary corrections regarding PAHs fluorescence and inner filter effect. From the numerical fitting the final association constants were calculated. Anthracene and anthraquinone failed to quench the albumin fluorescence. 9-anthracenecarboxylic acid showed the highest, while 9-anthracenemethanol the weakest albumin binding affinity. The affinity constants determined for 9-anthraldehyde and benzo[a]pyrene were of the same magnitude and indicated low-affinity binding to albumin. The constants obtained for the four dihydroxyanthraquinones were higher, but dissimilar, which suggests that the position of the functional group in anthracene molecule influences the binding constant. Moreover, this study suggests that the type of substituent plays a significant role in PAH-albumin complex formation. The carboxylic group increases the binding affinity of the anthracene molecule the most rather than the presence of both carbonyl and hydroxyl groups. The lowest affinity constants were obtained for aldehyde, methyl and carbonyl substituents.

4

Studies on nongenomic action by steroid hormones

100%

Kotwicka M. , Filipiak K. , Butowska W. , Warchol J. B.

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nr 1

5

Interaction of alkylresorcinols with proteins

100%

Kozubek A.

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tom 42

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nr 2

EN

Alkylresorcinol homologs form stable monomolecular layers at air-water interface. Their interaction with proteins present in the subphase results in an increase of alkylresorcinol molecular packing in the extent dependent upon the protein studied. Strongest effects were observed for proteins with large hydrophobic regions, e.g. glycophorin or serum albumin. Interaction of proteins with alkylresorcinol monolayers is stronger than with phospholipids. A decrease and a shift of intrinsic protein fluorescence upon interaction with the compounds studied support their involvement in alteration of hydrophobic regions. For trypsin, 50% quenching was observed at the alkylresorcinol/trypsin ratio of 0.75. Concomitantly, an apparent inhibition of the enzymatic activity was noted. These results indicate that direct interaction of alkylresorcinols and modulation of enzymatic activities should be recognised as a significant part of the biological effect of these cereal bran components.

6

Bacterial recognition of thermal glycation products derived from porcine serum albumin with lactose

80%

Sarabia-Sainz A. , Ramos-Clamont G. , Winzerling J. , Vazquez-Moreno L.

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tom 58

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nr 1

EN

Recently, glyco-therapy is proposed to prevent the interaction of bacterial lectins with host ligands (glycoconjugates). This interaction represents the first step in infection. Neoglycans referred to as PSA-Lac (PSA-Glu (β1-4) Gal) were obtained by conjugation of porcine serum albumin (PSA) with lactose at 80 °C, 100 °C and 120 ºC. Characterization studies of the products showed that PSA could contain 1, 38 or 41 added lactoses, depending on the reaction temperature. These neoglycans were approximately 10 times more glycated than PSA-Lac obtained in previous work. Lactose conjugation occurred only at lysines and PSA-Lac contained terminal galactoses as confirmed by Ricinus communis lectin recognition. Furthermore, Escherichia coli K88+, K88ab, K88ac and K88ad adhesins showed affinity toward all PSA-Lac neoglycans, and the most effective was the PSA-Lac obtained after 100 ºC treatment. In vitro, this neoglycan partially inhibited the adhesion of E. coli K88+ to piglet mucin (its natural ligand). These results provide support for the hypothesis that glycated proteins can be used as an alternative for bioactive compounds for disease prevention.

7

Antioxidants and environmental stress: spectroscopic study on stability of natural compounds and their interaction with a molecule of protein in an in vitro model

80%

Trnková L. , Boušová I. , Ryšánková L. , Vrabcová P. , Dršata J.

Proceedings of ECOpole

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2009

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tom Vol. 3, No. 1

27--34

EN

The stability of eight hydroxycinnamic acids (HCAs) during long-term incubation under physiological conditions was studied by UV-VIS absorption spectroscopy and their possibility of binding to a model protein (bovine serum albumin, BSA) under physiological conditions was investigated by tryptophan fluorescence quenching method. The obtained results suggest that the stability of hydroxycinnamic acids is dependent upon its individual structure and duration of incubation. The monosubstituted derivatives (coumaric acids) were stable within the course of long-term incubation, while di- and trisubstituted derivatives decomposed easily. It was found out that all studied compounds changed fluorescence emission spectrum of BSA. The Stern-Volmer analysis was employed in order to explore binding of HCAs to BSA in details. The binding constants (Kb), number of binding sites (n) and the free energy changes (∆G0) were determined. The binding affinity was strongest for rosmarinic acid and ranked in the following order rosmarinic acid > chlorogenic acid > sinapic acid > caffeic acid > ferulic acid > o-coumaric acid > p-coumaric acid > m-coumaric acid. All free energy changes (∆G0) possessed negative sign indicating the spontaneity of HCAs binding to BSA.

PL

Badano trwałość ośmiu kwasów hydrooksycynamonowych, HCAs, podczas inkubacji długoterminowej w warunkach fizjologicznych i możliwość ich wiązania się z modelowym białkiem (albuminą, BSA). Uzyskane wyniki pokazały, że trwałość tych kwasów zależy od ich indywidualnej budowy oraz czasu inkubacji. Wyznaczono stałe trwałości (Kb), liczbę miejsc wiązania (n) oraz zmianę wartości energii Gibbsa (∆G0) wiązania HCAs do BSA, który to proces przebiegał samorzutnie.

8

Immune regulation by sphingosine 1-phosphate and its receptors

80%

Bode C. , Graler M. H.

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nr 1

9

Influence of dendrimers on red blood cells

80%

Ziemba B. , Matuszko G. , Bryszewska M. , Klajnert B.

Cellular and Molecular Biology Letters

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2012

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tom 17

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nr 1

EN

The emerging fields of tissue engineering and biomaterials have begun to provide potential treatment options for liver failure. The goal of the present study is to investigate the ability of a poly L-lactic acid (PLLA) nanofiber scaffold to support and enhance hepatic differentiation of human bone marrow-derived mesenchymal stem cells (hMSCs). A scaffold composed of poly L-lactic acid and collagen was fabricated by the electrospinning technique. After characterizing isolated hMSCs, they were seeded onto PLLA nanofiber scaffolds and induced to differentiate into a hepatocyte lineage. The mRNA levels and protein expression of several important hepatic genes were determined using RT-PCR, immunocytochemistry and ELISA. Flow cytometry revealed that the isolated bone marrow-derived stem cells were positive for hMSC-specific markers CD73, CD44, CD105 and CD166 and negative for hematopoietic markers CD34 and CD45. The differentiation of these stem cells into adipocytes and osteoblasts demonstrated their multipotency. Scanning electron microscopy showed adherence of cells in the nanofiber scaffold during differentiation towards hepatocytes. Our results showed that expression levels of liver-specific markers such as albumin, α-fetoprotein, and cytokeratins 8 and 18 were higher in differentiated cells on the nanofibers than when cultured on plates. Importantly, liver functioning serum proteins, albumin and α-1 antitrypsin were secreted into the culture medium at higher levels by the differentiated cells on the nanofibers than on the plates, demonstrating that our nanofibrous scaffolds promoted and enhanced hepatic differentiation under our culture conditions. Our results show that the engineered PLLA nanofibrous scaffold is a conducive matrix for the differentiation of MSCs into functional hepatocyte-like cells. This represents the first step for the use of this nanofibrous scaffold for culture and differentiation of stem cells that may be employed for tissue engineering and cell-based therapy applications.

10

Effect of the combination of acetylsalicylic acid or chlormethine with antibiotics and glucose on the mammary gland of healthy cows

60%

Malinowski E.

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1992

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tom 36