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1.
Janine F. Bridges Peter C. Millard John F. Woodley 《Biochimica et Biophysica Acta (BBA)/General Subjects》1978,544(2):448-451
The uptake of free and liposome-entrapped 125I-labelled poly(vinylpyrrolidone) was measured in an intestinal sac preparation from adult rats. An an equal concentration of 125I-labelled poly(vinylpyrrolidone), the rate of uptake of the liposome-entrapped material was four times that of the free macromolecule. 相似文献
2.
4-Thiaproline reduces heart lipid peroxidation and collagen accumulation in the diabetic db/db mouse
Summary Collagen accumulation is a main pathological feature of diabetic cardiomyopathy. The underlying mechanisms seem to be increased cross linking by reactive carbonyles. The purpose of the study was to decrease the collagen content of total ventricular tissue by the oral administration of thiaproline, which could reduce collagen due to its functions as a proline analogue, blocking collagen production and as a free oxygen radical scavenger, blocking reactive carbonyles and oxygen species and subsequently collagen cross linking.Thiaproline was administered to genetically diabetic db/db mice and compared to untreated animals. Total ventricular collagen as expressed by hydroxyproline was significantly lower in the treated group (means 0.23 micromoles/10 tissue in the treated vs 0.35 micromoles/100 mg tissue in the untreated group, p < 0.001). Significantly more collagen could be eluted in the treated group (p < 0.001) and carboxymethyllysine was significantly reduced in the treated group (p < 0.001). Di-tyrosine and glycemic control did not differ between the groups. Glutathione was significantly increased in the TP treated experimental group (p < 0.001) and lipid peroxidation products were significantly decreased (means 0.221 absorbance in the treated group versus 0.321 absorbance in the untreated diabetic group) correlating with total ventricular collagen content (r = 0.87, p < 0.01).We conclude that thiaproline reduced total ventricular collagen content by inhibiting collagen cross linking as reflected by increased solubility of collagen and expressed by higher elution quantity of collagen. Thiaproline, and/or its metabolites induced increase of heart glutathione which may well have been scavenging reactive carbonyles derived from lipid peroxidation and advanced stage nonenzymatic glycosylation as shown by decreased total ventricular carboxy-methyllysine and lipid peroxidation products paralleling reduced heart collagen content.It remains to be shown that the successful reduction of heart collagen by thiaproline is paralleled by improved functional properties. 相似文献
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Jacob L. Jordan Joseph W. Arndt Karl Hanf Guohui Li Janine Hall Stephen Demarest Flora Huang Xiufeng Wu Brian Miller Scott Glaser Erik J. Fernandez Deping Wang Alexey Lugovskoy 《Proteins》2009,77(4):832-841
Bispecific immunoglobulin‐like antibodies capable of engaging multiple antigens represent a promising new class of therapeutic agents. Engineering of these molecules requires optimization of the molecular properties of one of the domain components. Here, we present a detailed crystallographic and computational characterization of the stabilization patterns in the lymphotoxin‐beta receptor (LTβR) binding Fv domain of an anti‐LTβR/anti‐TNF‐related apoptosis inducing ligand receptor‐2 (TRAIL‐R2) bispecific immunoglobulin‐like antibody. We further describe a new hierarchical structure‐guided approach toward engineering of antibody‐like molecules to enhance their thermal and chemical stability. Proteins 2009. © 2009 Wiley‐Liss, Inc. 相似文献
5.
6.
Ravinder Kaur Grewal Monika Lulsdorf Janine Croser Sergio Ochatt Albert Vandenberg Thomas D. Warkentin 《Plant cell reports》2009,28(8):1289-1299
This is the first report on the production of double-haploid chickpea embryos and regenerated plants through anther culture
using Canadian cultivar CDC Xena (kabuli) and Australian cultivar Sonali (desi). Maximum anther induction rates were 69% for
Sonali and 63% for CDC Xena. Under optimal conditions, embryo formation occurred within 15–20 days of culture initiation with
2.3 embryos produced per anther for CDC Xena and 2.0 embryos per anther for Sonali. For anther induction, the following stress
treatments were used: (1) flower clusters were treated at 4°C for 4 days, (2) anthers were subjected to electric shock treatment
of three exponentially decaying pulses of 50–400 V with 25 μF capacitance and 25 Ω resistance, (3) anthers were centrifuged
at 168–1,509g for 2–15 min, and finally (4) anthers were cultured for 4 days in high-osmotic pressure (563 mmol) liquid medium. Anthers
were then transferred to a solid embryo development medium and, 15–20 days later, embryo development was observed concomitant
with a small amount of callus growth of 0.1–3 mm. Anther-derived embryos were regenerated on plant regeneration medium. Electroporation
treatment of anthers enhanced root formation, which is often a major hurdle in legume regeneration protocols. Cytological
studies using DAPI staining showed a wide range of ploidy levels from haploid to tetraploid in 10–30-day-old calli. Flow cytometric
analysis of calli, embryos and regenerated plants showed haploid profiles and/or spontaneous doubling of the chromosomes during
early regeneration stages. 相似文献
7.
Hanania EG Fieck A Stevens J Bodzin LJ Palsson BØ Koller MR 《Biotechnology and bioengineering》2005,91(7):872-876
Cloning of highly-secreting recombinant cells is critical for biopharmaceutical manufacturing, but faces numerous challenges including the fact that secreted protein does not remain associated with the producing cell. A fundamentally new approach was developed combining in situ capture and measurement of individual cell protein secretion followed by laser-mediated elimination of all non- and poorly-secreting cells, leaving only the highest-secreting cell in a well. Recombinant cells producing humanized antibody were cultured serum-free on a capture matrix, followed by staining with fluorescently-labeled anti-human antibody fragment. A novel, automated, high-throughput instrument (called LEAP) was used to image and locate every cell, quantify the cell-associated and secreted antibody (surrounding each cell), eliminate all undesired cells from a well via targeted laser irradiation, and then track clone outgrowth and stability. Temporarily sparing an island of helper cells around the clone of interest improved cloning efficiency (particularly when using serum-free medium), and helper cells were easily eliminated with the laser after several days. The in situ nature of this process allowed several serial sub-cloning steps to be performed within days of one another, resulting in rapid generation of clonal populations with significantly increased and more stable, homogeneous antibody secretion. Cell lines with specific antibody secretion rates of > 50 pg/cell per day (in static batch culture) were routinely obtained as a result of this cloning approach, often times representing up to 20% of the clones screened. 相似文献
8.
Procópio L Alvarez VM Jurelevicius DA Hansen L Sørensen SJ Cardoso JS Pádula M Leitão AC Seldin L van Elsas JD 《Antonie van Leeuwenhoek》2012,101(2):289-302
The draft genome of Dietzia cinnamea strain P4 was determined using pyrosequencing. In total, 428 supercontigs were obtained and analyzed. We here describe and
interpret the main features of the draft genome. The genome contained a total of 3,555,295 bp, arranged in a single replicon
with an average G+C percentage of 70.9%. It revealed the presence of complete pathways for basically all central metabolic
routes. Also present were complete sets of genes for the glyoxalate and reductive carboxylate cycles. Autotrophic growth was
suggested to occur by the presence of genes for aerobic CO oxidation, formate/formaldehyde oxidation, the reverse tricarboxylic
acid cycle and the 3-hydropropionate cycle for CO2 fixation. Secondary metabolism was evidenced by the presence of genes for the biosynthesis of terpene compounds, frenolicin,
nanaomycin and avilamycin A antibiotics. Furthermore, a probable role in azinomycin B synthesis, an important product with
antitumor activity, was indicated. The complete alk operon for the degradation of n-alkanes was found to be present, as were clusters of genes for biphenyl ring dihydroxylation. This study brings new insights
in the genetics and physiology of D. cinnamea P4, which is useful in biotechnology and bioremediation. 相似文献
9.
β-Ketoacyl-ACP synthase (KAS) enzymes catalyze Claisen condensation reactions in the fatty acid biosynthesis pathway. These reactions follow a ping-pong mechanism in which a donor substrate acylates the active site cysteine residue after which the acyl group is condensed with the malonyl-ACP acceptor substrate to form a β-ketoacyl-ACP. In the priming KASIII enzymes the donor substrate is an acyl-CoA while in the elongating KASI and KASII enzymes the donor is an acyl-ACP. Although the KASIII enzyme in Escherichia coli (ecFabH) is essential, the corresponding enzyme in Mycobacterium tuberculosis (mtFabH) is not, suggesting that the KASI or II enzyme in M. tuberculosis (KasA or KasB, respectively) must be able to accept a CoA donor substrate. Since KasA is essential, the substrate specificity of this KASI enzyme has been explored using substrates based on phosphopantetheine, CoA, ACP, and AcpM peptide mimics. This analysis has been extended to the KASI and KASII enzymes from E. coli (ecFabB and ecFabF) where we show that a 14-residue malonyl-phosphopantetheine peptide can efficiently replace malonyl-ecACP as the acceptor substrate in the ecFabF reaction. While ecFabF is able to catalyze the condensation reaction when CoA is the carrier for both substrates, the KASI enzymes ecFabB and KasA have an absolute requirement for an ACP substrate as the acyl donor. Provided that this requirement is met, variation in the acceptor carrier substrate has little impact on the k(cat)/K(m) for the KASI reaction. For the KASI enzymes we propose that the binding of ecACP (AcpM) results in a conformational change that leads to an open form of the enzyme to which the malonyl acceptor substrate binds. Finally, the substrate inhibition observed when palmitoyl-CoA is the donor substrate for the KasA reaction has implications for the importance of mtFabH in the mycobacterial FASII pathway. 相似文献
10.
Sara Rosati Ewald TJ van den Bremer Janine Schuurman Paul WHI Parren Johannis P Kamerling Albert JR Heck 《MABS-AUSTIN》2013,5(6):917-924
Here, we describe a fast, easy-to-use, and sensitive method to profile in-depth structural micro-heterogeneity, including intricate N-glycosylation profiles, of monoclonal antibodies at the native intact protein level by means of mass spectrometry using a recently introduced modified Orbitrap Exactive Plus mass spectrometer. We demonstrate the versatility of our method to probe structural micro-heterogeneity by describing the analysis of three types of molecules: (1) a non-covalently bound IgG4 hinge deleted full-antibody in equilibrium with its half-antibody, (2) IgG4 mutants exhibiting highly complex glycosylation profiles, and (3) antibody-drug conjugates. Using the modified instrument, we obtain baseline separation and accurate mass determination of all different proteoforms that may be induced, for example, by glycosylation, drug loading and partial peptide backbone-truncation. We show that our method can handle highly complex glycosylation profiles, identifying more than 20 different glycoforms per monoclonal antibody preparation and more than 30 proteoforms on a single highly purified antibody. In analyzing antibody-drug conjugates, our method also easily identifies and quantifies more than 15 structurally different proteoforms that may result from the collective differences in drug loading and glycosylation. The method presented here will aid in the comprehensive analytical and functional characterization of protein micro-heterogeneity, which is crucial for successful development and manufacturing of therapeutic antibodies 相似文献