An albumin-derived peptide scaffold capable of binding and catalysis

We have identified a 101-amino-acid polypeptide derived from the sequence surrounding the IIA binding site of human albumin. The polypeptide contains residues that make contact with ligands as warfarin in the parent protein, and eight cysteine residues to form disulfide bridges, which stabilize the polypeptide structure. Seventy-four amino acids are located in six [alpha]-helical regions, with the remaining amino acids forming six connecting coil/loop regions. Codon usage optimization was used to express a GST fusion protein in E. coli in yields as high as 4 mg/l. This fusion protein retains its structural integrity and aldolase activity, the ability to direct the stereochemical outcome of a diketone reduction, and its binding capacity to warfarin and efavirenz. Notably, this newly cloned polypeptide represents a valuable starting point for the construction of libraries of binders and catalysts with improved proficiency

Plasma α-tocopherol content and its relationship with milk somatic cells count in Italian commercial herds.

This work was aimed to investigate relationship between plasma vitamin E concentration and milk somatic cell count in healthy cows in commercial herds. 49 multiparous cows from two commercial dairy herds were monitored from the day of dry off until 90 DIM. BCS was assessed and blood samples were collected at dry off, day 0, 30, 60 and 90 postpartum. Plasma was analyzed for α-tocopherol content. Quantification of NEFA, BOHB, Zn and Se was performed in serum samples. Milk production and composition was obtained from routinely test-day of Italian milk producers association. Somatic Cell Score (SCS) was calculated and included in the dataset. Analysis of data was performed using MIXED repeated and CORR procedures of SAS.We did not observe a correlation between plasmatic vitamin E and somatic cell score, and this can be explained by the low level of somatic cell score (averages 1.64 and 1.26). The lowest value of vitamin E was observed at parturition (1.64 µg/ml and 1.95 µg/ml). A significant (P<0.01) negative (-20%) correlation was observed between NEFA serum content and α-tocopherol plasma concentration. Serum selenium content was positively correlated (+42%, P<0.0001) to zinc concentration. Grouping cows on the basis of their plasma α-tocopherol content higher or lower than 3 μg/mL at dry off, SCS at 30 and 60 DIM tended to be higher in lactating animals with lower content of α-tocopherol (1.12 vs. 1.72, P=0.18 at 30d; 0.92 vs. 1.72, P=0.07 at 60d). However, plasma α-tocopherol content at dry off could be usefully correlated with somatic cell count in fresh cows

Definition of technology readiness levels (trl) and application to biomedical field

Dedicatoria: "Dedicado al Exmo Sr D. Lorenzo Arrazola, Ministro que ha sido cuatro veces de la Corona, Presidente del Tribunal Supremo de Justicia, Senador..."Coordenadas referidas al meridiano de Madrid (O 7°20'--E 9°45'/N 44°00'--N 35°00'). Red geográfica de 1° en 1°Relieve representado por curvas de configuraciónTabla de signos convencionales para indicar Audiencias, Partido judicial de término, de ascenso y de entrada, poblaciones con dos o más juzgados, presidios, casas correccionales de mujeres y división por audiencias, provincias y paísesEn el margen izquierdo consta una leyenda con los jurisconsultos agrupados por provincia

Morphological and molecular modifications induced by heat shock in <i>Drosophila melanogaster</i> embryos

ABSTRACT The early embryo of Drosophila melanogaster did not survive treatment at 37°C (heat shock) for 25 min. The histological analysis of eggs treated in this way showed that the heat shock caused disintegration of nuclei and of cytoplasmic islands, displacement and swelling of nuclei and blocked mitoses. These effects were not observed in embryos treated after blastoderm formation. After this stage, we noticed that development was slowed down. The heat shock proteins (hsp 83,70 and 68) were, under shock, synthesized at all developmental stages. There was little or no synthesis of hsp 70 and 68 in unfertilized eggs, but synthesis increased in proportion to the number of nuclei present. Most probably, hsp 70 synthesis was directed by zygotic mRNA. DNA synthesis was not blocked by the heat shock though the overall incorporation of pH]thymidine was substantially reduced, presumably because of the block of mitoses. We did not find a direct relation between survival pattern and hsp synthesis. We concluded that some, at least, of the heat shock genes can be activated at all developmental stages and that heat shock could be used for synchronizing mitoses.</jats:p

Designing Short Peptides with High Affinity for Organic Molecules: A Combined Docking, Molecular Dynamics and Monte Carlo Approach

We present a method for designing artificial receptors capable of binding with high affinity to a chosen target organic molecule. The primary sequence of the peptide is optimized to maximize its binding affinity. Our algorithm builds on a combination of molecular dynamics, semiflexible docking, and replica exchange Monte Carlo and performs simultaneous sampling in sequence and conformational spaces carefully selecting the degree of flexibility in the mutated peptides. The approach is used to design a decapeptide able to bind efavirenz. The calculated binding energy of the designed peptide (approximately −12 kcal/mol) was confirmed experimentally by fluorescence measurements. NMR spectroscopy confirmed the interactions between the peptide and the efavirenz molecule predicted by the algorithm