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31.
Sabrina Ait Braham Fouzia Hussain Roberto Morellon-Sterling Shagufta Kamal Jakub F. Kornecki Oveimar Barbosa Djamel Edine Kati Roberto Fernandez-Lafuente 《Biotechnology progress》2019,35(2):e2768
Alcalase was scarcely immobilized on monoaminoethyl-N-aminoethyl (MANAE)-agarose beads at different pH values (<20% at pH 7). The enzyme did not immobilize on MANAE-agarose activated with glutaraldehyde at high ionic strength, suggesting a low reactivity of the enzyme with the support functionalized in this manner. However, the immobilization is relatively rapid when using low ionic strength and glutaraldehyde activated support. Using these conditions, the enzyme was immobilized at pH 5, 7, and 9, and in all cases, the activity vs. Boc-Ala-ONp decreased to around 50%. However, the activity vs. casein greatly depends on the immobilization pH, while at pH 5 it is also 50%, at pH 7 it is around 200%, and at pH 9 it is around 140%. All immobilized enzymes were significantly stabilized compared to the free enzyme when inactivated at pH 5, 7, or 9. The highest stability was always observed when the enzyme was immobilized at pH 9, and the worst stability occurred when the enzyme was immobilized at pH 5, in agreement with the reactivity of the amino groups of the enzyme. Stabilization was lower for the three preparations when the inactivation was performed at pH 5. Thus, this is a practical example on how the cooperative effect of ion exchange and covalent immobilization may be used to immobilize an enzyme when only one independent cause of immobilization is unable to immobilize the enzyme, while adjusting the immobilization pH leads to very different properties of the final immobilized enzyme preparation. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2768, 2019. 相似文献
32.
Anton GT Terwisscha van Scheltinga Marjolijn N Lub-de Hooge Keelara Abiraj Carolien P Schr?der Linda Pot Birgit Bossenmaier Marlene Thomas Gabriele H?lzlwimmer Thomas Friess Jos GW Kosterink Elisabeth GE de Vries 《MABS-AUSTIN》2014,6(4):1051-1058
The humanized monoclonal antibody with high affinity for the human epidermal growth factor receptor (HER) 3, RG7116, is a glycoengineered, IgG1 class antibody. By labeling RG7116 with zirconium-89 (89Zr) we aimed to visualize in vivo HER3 expression and study the biodistribution of this antibody in human tumor-bearing mice. Biodistribution of 89Zr-RG7116 was studied in subcutaneously xenografted FaDu tumor cells (HER3-positive). Dose-dependency of 89Zr-RG7116 organ distribution and specific tumor uptake was assessed by administering doses ranging from 0.05 to 10 mg/kg RG7116 to SCID/Beige mice. Biodistribution was analyzed at 24 and 144 h after injection. MicroPET imaging was performed at 1, 3, and 6 days after injection of 1.0 mg/kg 89Zr-RG7116 in the FaDu, H441, QG-56 and Calu-1 xenografts with varying HER3 expression. The excised tumors were analyzed for HER3 expression. Biodistribution analyses showed a dose- and time-dependent 89Zr-RG7116 tumor uptake in FaDu tumors. The highest tumor uptake of 89Zr-RG7116 was observed in the 0.05 mg/kg dose group with 27.5%ID/g at 144 h after tracer injection. MicroPET imaging revealed specific tumor uptake of 89Zr-RG7116 in FaDu and H441 models with an increase in tumor uptake over time. Biodistribution data was consistent with the microPET findings in FaDu, H441, QG56 and Calu-1 xenografts, which correlated with HER3 expression levels. In conclusion, 89Zr-RG7116 specifically accumulates in HER3 expressing tumors. PET imaging with this tracer provides real-time non-invasive information about RG7116 distribution, tumor targeting and tumor HER3 expression levels. 相似文献
33.
Braham Parkash Atul Ranjan Vinod Tiwari Sharad Kumar Gupta Navrinder Kaur Vibha Tandon 《PloS one》2012,7(11)
Background
The genome of retroviruses, including HIV-1, is packaged as two homologous (+) strand RNA molecules, noncovalently associated close to their 5′-end in a region called dimer linkage structure (DLS). Retroviral HIV-1 genomic RNAs dimerize through complex interactions between dimerization initiation sites (DIS) within the (5′-UTR). Dimer formation is prevented by so calledLong Distance Interaction (LDI) conformation, whereas Branched Multiple Hairpin (BMH) conformation leads to spontaneous dimerization.Methods and Results
We evaluated the role of SL1 (DIS), PolyA Hairpin signal and a long distance U5-AUG interaction by in-vitro dimerization, conformer assay and coupled dimerization and template-switching assays using antisense PNAs. Our data suggests evidence that PNAs targeted against SL1 produced severe inhibitory effect on dimerization and template-switching processes while PNAs targeted against U5 region do not show significant effect on dimerization and template switching, while PNAs targeted against AUG region showed strong inhibition of dimerization and template switching processes.Conclusions
Our results demonstrate that PNA can be used successfully as an antisense to inhibit dimerization and template switching process in HIV -1 and both of the processes are closely linked to each other. Different PNA oligomers have ability of switching between two thermodynamically stable forms. PNA targeted against DIS and SL1 switch, LDI conformer to more dimerization friendly BMH form. PNAs targeted against PolyA haipin configuration did not show a significant change in dimerization and template switching process. The PNA oligomer directed against the AUG strand of U5-AUG duplex structure also showed a significant reduction in RNA dimerization as well as template- switching efficiency.The antisense PNA oligomers can be used to regulate the shift in the LDI/BMH equilibrium. 相似文献34.
Besma Lakhal Sonia Ben-Hadj-Khalifa Nouha Bouali Pascal Philipert Françoise Audran Rim Braham Elghezal Hatem Charles Sultan Ali Saad 《Gene》2012
Background
WNT4 and SF1 genes play an important role in ovarian development. They constitute coherent candidate genes associated with premature ovarian failure (POF) pathogenesis.Methods
We sequenced the coding region of WNT4 and SF1 in 55 Tunisian women with POF and 100 healthy controls.Results
We identified a synonymous variation in WNT4 (c.99G>A, p.Ser33Ser) and a substitution (c.G437C) in SF1 gene inducing G146 to Ala (GGG–GCG) missense mutation. WNT4 (c.99G>A, p.Ser33Ser) was not associated with POF pathology. However, a positive association of SF1 Gly146Ala polymorphism was noted. Gly146Ala minor allele frequency was significantly higher (p = 0.029) in POF patients versus controls and Ala allele containing genotypes (p = 0.005) were positively associated with POF pathology. The carriage of 146Ala allele was also associated with a significant reduction in estradiol plasma levels.Conclusions
SF1 Gly146Ala polymorphism seems to be associated with POF pathology in the Tunisian population likely by reducing estradiol levels. 相似文献35.
Seed Bank Viability in Disturbed Longleaf Pine Sites 总被引:4,自引:0,他引:4
Some of the most species‐rich areas and highest concentrations of threatened and endangered species in the southeastern United States are found in wet savanna and flatwood longleaf pine (Pinus palustris Mill.) communities. Where intensive forestry practices have eliminated much of the natural understory of the longleaf ecosystem, the potential for reestablishment through a seed bank may present a valuable restoration opportunity. Longleaf pine sites converted to loblolly pine plantations and non‐disturbed longleaf sites on the Coastal Plain of North Carolina were examined for seed bank presence and diversity. Conducting vegetation surveys and examining the seed bank using the seedling emergence technique allowed for verification of the seed bank presence, as well as evaluation of the quality of the seed bank on disturbed longleaf pine sites. Forty‐three species and over 1,000 individuals germinated, and the seed banks of both the disturbed and non‐disturbed stand types contained species not noted in the vegetation survey. Although many of these species were considered weedy and typical of disturbance, numerous taxa were indicative of stable longleaf pine communities. This study confirms both the presence and quality of seed banks in highly disturbed former longleaf pine sites, suggesting that the seed bank may be an important tool in restoration efforts. 相似文献
36.
Brian W. Rolek Melissa A. Braham Tricia A. Miller Adam E. Duerr Todd E. Katzner Christopher J. W. McClure 《Ibis》2024,166(1):308-314
Automated curtailment of wind turbines can reduce fatality rates of wildlife but the resulting increased number of curtailments can reduce power generation. Tailoring curtailment criteria for each individual turbine could reduce unnecessary curtailment, yet it is unknown whether the risk to wildlife varies among turbines. We demonstrate turbine-specific variation in the speed, altitude, approach angle and distance metrics associated with entry by eagles into rotor-swept zones. Our results thus illustrate the potential value of turbine-specific curtailment criteria to reduce fatality rates of wildlife at wind energy facilities. 相似文献
37.
Hypothesis for a serine proteinase-like domain at the COOH terminus of Slowpoke calcium-activated potassium channels 总被引:5,自引:4,他引:1
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Bovine pancreatic trypsin inhibitor (BPTI) is a 58-residue protein with three disulfide bonds that belongs to the Kunitz family of serine proteinase inhibitors. BPTI is an extremely potent inhibitor of trypsin, but it also specifically binds to various active and inactive serine proteinase homologs with KD values that range over eight orders of magnitude. We previously described an interaction of BPTI at an intracellular site that results in the production of discrete subconductance events in large conductance Ca2+ activated K+ channels (Moss, G.W.J., and E. Moczydlowski. 1996, J. Gen. Physiol, 107:47-68). In this paper, we summarize a variety of accumulated evidence which suggests that BPTI binds to a site on the KCa channel protein that structurally resembles a serine proteinase. One line of evidence includes the finding that the complex of BPTI and trypsin, in which the inhibitory loop of BPTI is masked by interaction with trypsin, is completely ineffective in the production of substate events in the KCa channel. To further investigate this notion, we performed a sequence analysis of the alpha-subunit of cloned slowpoke KCa channels from Drosophila and mammals. This analysis suggests that a region of approximately 250 residues near the COOH terminus of the KCa channel is homologous to members of the serine proteinase family, but is catalytically inactive because of various substitutions of key catalytic residues. The sequence analysis also predicts the location of a Ca(2+)-binding loop that is found in many serine proteinase enzymes. We hypothesize that this COOH-terminal domain of the slowpoke KCa channel adopts the characteristic double-barrel fold of serine proteinases, is involved in Ca(2+)-activation of the channel, and may also bind other intracellular components that regulate KCa channel activity. 相似文献
38.
B. Chermiti M. Braham M. Znaidi H. Gahbiche B. Messelmani M. Dali & H. Messelmani 《Journal of Applied Entomology》2001,125(1-2):45-52
First results on releasing experiments of Ageniaspis citricola Logvinovskaya (Hym., Encyrtidae), a parasitoid of Phyllocnistis citrella Stainton (Lep., Gracillariidae) in Tunisia . Trials were conducted to reduce the population of the citrus leafminer Phyllocnistis citrella in Citrus orchards by releasing the imported specific parasitoid Ageniaspis citricola in Tunisia. Adults of the parasitoid were released in 69 orchards between 1 June and 23 December 1996, corresponding to the two main flushes of Citrus . Results indicate that the parasitoid was established in 27 orchards reaching a maximum percentage of parasitism of 28%. In 1997, A. citricola is established in some Citrus groves proving its real acclimatization in Tunisia, with a maximum percentage of parasitism reaching 45%. 相似文献
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