Migrations of yellowfin tuna tagged off the southern coast of Mexico in 1960 and 1969

ENGLISH: The map method, the Jones method, the variance-covariance method, and the Skellam method were used to study the migrations of tagged yellowfin tuna released off the southern coast of Mexico in 1960 and 1969. The first three methods are all useful, and each presents information which is complementary to that presented by the others. The Skellam method, as used in this report, is less useful. The movements of the tagged fish released in 1960 appeared to have been strongly directed, but this was probably caused principally by the distribution of the fishing effort. The effort was much more widely distributed in 1970, and the movements of the fish released in 1969 appeared to have been much less directed. The correlation coefficients derived from the variance-covariance method showed that it was not random, however. The small fish released in the Acapulco and 10°N-100°W areas in 1969 migrated to the Manzanillo area near the beginning of February 1970. The medium and large fish released in the same areas in the same year tended to migrate to the southeast throughout the first half of 1970, however. SPANISH: El método de mapas, el de Jones, el de la variancia-covariancia y el de Skellam fueron empleados para estudiar las migraciones del atún aleta amarilla marcado y liberado frente a la costa meridional de México en 1960 y 1969. Los tres primeros métodos son todos útiles, y cada uno presenta información que complementa la presentada por los otros. El método de Skellam, conforme se usa en este informe, es menos útil. Parece que los desplazamientos de los peces marcados y liberados en 1960 hubieran sido fuertemente orientados, pero ésto probablemente fue causado principalmente por la distribución del esfuerzo de pesca. El esfuerzo se distribuyó más extensamente en 1970, y parece que los desplazamientos de los peces liberados en 1969 fueran menos orientados. Los coeficientes de correlación derivados del método variancia-covariancia indicaron, sin embargo, que no eran aleatorios. Los peces pequeños liberados en las áreas de Acapulco y los 10°N-100°W en 1969 migraron al área de Manzanillo a principios de febrero 1970. Los peces medianos y grandes liberados en las mismas áreas en el mismo año tuvieron, sin embargo, la tendencia a desplazarse al sudeste durante el primer semestre de 1970. (PDF contains 64 pages.)

Functional analysis of cancer-associated EGFR mutants using a cellular assay with YFP-tagged EGFR intracellular domain

Background: The presence of EGFR kinase domain mutations in a subset of NSCLC patients correlates with the response to treatment with the EGFR tyrosine kinase inhibitors gefitinib and erlotinib. Although most EGFR mutations detected are short deletions in exon 19 or the L858R point mutation in exon 21, more than 75 different EGFR kinase domain residues have been reported to be altered in NSCLC patients. The phenotypical consequences of different EGFR mutations may vary dramatically, but the majority of uncommon EGFR mutations have never been functionally evaluated. Results: We demonstrate that the relative kinase activity and erlotinib sensitivity of different EGFR mutants can be readily evaluated using transfection of an YFP-tagged fragment of the EGFR intracellular domain (YFP-EGFR-ICD), followed by immunofluorescence microscopy analysis. Using this assay, we show that the exon 20 insertions Ins770SVD and Ins774HV confer increased kinase activity, but no erlotinib sensitivity. We also show that, in contrast to the common L858R mutation, the uncommon exon 21 point mutations P848L and A859T appear to behave like functionally silent polymorphisms. Conclusion: The ability to rapidly obtain functional information on EGFR variants of unknown relevance using the YFP-EGFR-ICD assay might prove important in the future for the management of NSCLC patients bearing uncommon EGFR mutations. In addition, our assay may be used to determine the response of resistant EGFR mutants to novel second-generation TKIs.

Molecular Modeling and Affinity Determination of scFv Antibody: Proper Linker Peptide Enhances Its Activity

One of existing strategies to engineer active antibody is to link VH and VL domains via a linker peptide. How the composition, length, and conformation of the linker affect antibody activity, however, remains poorly understood. In this study, a dual approach that coordinates molecule modeling, biological measurements, and affinity evaluation was developed to quantify the binding activity of a novel stable miniaturized anti-CD20 antibody or singlechain fragment variable (scFv) with a linker peptide. Upon computer-guided homology modeling, distance geometry analysis, and molecular superimposition and optimization, three new linker peptides PT1, PT2, and PT3 with respective 7, 10, and 15 residues were proposed and three engineered antibodies were then constructed by linking the cloned VH and VL domains and fusing to a derivative of human IgG1. The binding stability and activity of scFv-Fc chimera to CD20 antigen was quantified using a micropipette adhesion frequency assay and a Scatchard analysis. Our data indicated that the binding affinity was similar for the chimera with PT2 or PT3 and ~24-fold higher than that for the chimera with PT1, supporting theoretical predictions in molecular modeling. These results further the understanding in the impact of linker peptide on antibody structure and activity.

Developing peptide based capture agents for diagnostics and therapeutics

Iterative in situ click chemistry (IISCC) is a robust general technology for development of high throughput, inexpensive protein detection agents. In IISCC, the target protein acts as a template and catalyst, and assembles its own ligand from modular blocks of peptides. This process of ligand discovery is iterated to add peptide arms to develop a multivalent ligand with increased affinity and selectivity. The peptide based protein capture agents (PCC) should ideally have the same degree of selectivity and specificity as a monoclonal antibody, along with improved chemical stability. We had previously reported developing a PCC agent against bovine carbonic anhydrase II (bCAII) that could replace a polyclonal antibody. To further enhance the affinity or specificity of the PCC agent, I explore branching the peptide arms to develop branched PCC agents against bCAII. The developed branched capture agents have two to three fold higher affinities for the target protein. In the second part of my thesis, I describe the epitope targeting strategy, a strategy for directing the development of a peptide ligand against specific region or fragment of the protein. The strategy is successfully demonstrated by developing PCC agents with low nanomolar binding affinities that target the C-terminal hydrophobic motif of Akt2 kinase. One of the developed triligands inhibits the kinase activity of Akt. This suggests that, if targeted against the right epitope, the PCC agents can also influence the functional properties of the protein. The exquisite control of the epitope targeting strategy is further demonstrated by developing a cyclic ligand against Akt2. The cyclic ligand acts as an inhibitor by itself, without any iteration of the ligand discovery process. The epitope targeting strategy is a cornerstone of the IISCC technology and opens up new opportunities, leading to the development of protein detection agents and of modulators of protein functions.

Identification of thermally-tagged coherent structures in the zero pressure gradient turbulent boundary layer

A zero pressure gradient boundary layer over a flat plate is subjected to step changes in thermal condition at the wall, causing the formation of internal, heated layers. The resulting temperature fluctuations and their corresponding density variations are associated with turbulent coherent structures. Aero-optical distortion occurs when light passes through the boundary layer, encountering the changing index of refraction resulting from the density variations. Instantaneous measurements of streamwise velocity, temperature and the optical deflection angle experienced by a laser traversing the boundary layer are made using hot and cold wires and a Malley probe, respectively. Correlations of the deflection angle with the temperature and velocity records suggest that the dominant contribution to the deflection angle comes from thermally-tagged structures in the outer boundary layer with a convective velocity of approximately 0.8U∞. An examination of instantaneous temperature and velocity and their temporal gradients conditionally averaged around significant optical deflections shows behavior consistent with the passage of a heated vortex. Strong deflections are associated with strong negative temperature gradients, and strong positive velocity gradients where the sign of the streamwise velocity fluctuation changes. The power density spectrum of the optical deflections reveals associated structure size to be on the order of the boundary layer thickness. A comparison to the temperature and velocity spectra suggests that the responsible structures are smaller vortices in the outer boundary layer as opposed to larger scale motions. Notable differences between the power density spectra of the optical deflections and the temperature remain unresolved due to the low frequency response of the cold wire.

A biologically active peptide in the skin of lampreys (Eudontomyzon danfordi vladykovi). [Translation from: Z.Naturforsch. (B), 26(10), 1021-1023, 1971. ]

In recent years interest in the production and description of kinin-type substances has been greatly intensified. So, for example, bradykinin, phyllokinin, physalaemin, ranatensin and caerulein could be extracted from the skin of amphibians as well as. eledoisin out of the salivary glands of Eledon moschata. An examination of lampreys seemed to us particularly profitable in the search for the incidence of further kinins. Ammocoetes of different sizes and also adults of both sexes of the species Eudontomyzon danfordi vladykovi were studied in this research. This species is found in many tributaries of the Danube. Skin extracts were tested on on isolated rat uterus, rat duodenum, guinea pig ileum and rabbit jejunum, further tests were done in order to determine a peptide character of the biologically active substance.

The structure specificity of alpha-chymotrypsin. I. Polypeptides as substrates. II. N-acylated peptide esters as substrates. III. Some reactive esters of N-acylated amino acids as substrates

The structural specificity of α-chymotrypsin for polypeptides and denatured proteins has been examined. The primary specificity of the enzyme for these natural substrates is shown to closely correspond to that observed for model substrates. A pattern of secondary specificity is proposed.

A series of N-acetylated peptide esters of varying length have been evaluated as substrates of α-chymotrypsin. The results are interpreted in terms of proposed specificity theories.

The α-chymotrypsin-catalyzed hydrolyses of a number of N-acetylated dipeptide methyl esters were studied. The results are interpreted in terms of the available specificity theories and are compared with results obtained in the study of polypeptide substrates. The importance of non-productive binding in determining the kinetic parameters of these substrates is discussed. A partial model of the locus of the active site which interacts with the R’₁CONH- group of a substrate of the form R’₁CONHCHR₂COR’₃ is proposed.

Finally, some reactive esters of N-acetylated amino acids have been evaluated as substrates of α-chymotrypsin. Their reactivity and stereo-chemical behavior are discussed in terms of the specificity theories available. The importance of a binding interaction between the carboxyl function of the substrate and the enzyme is suggested by the results obtained.

I. Nuclear magnetic resonance analysis of ferrocenylcarbonium ion. II. Ferrocene catalyzed photochemistry

I. Nuclear magnetic resonance spectra of appropriately substituted ferrocenylcarbonium ions reveal the α-protons of the substituted ring to be more shielded than β-protons. The observation is discussed in terms of various models proposed for the ferrocenylcarbonium ion and is found to support a model in which the iron is bonded to all six carbona of the substituted ring.

II. Ferrocene catalyzes the photoisomerization of the piperylenes and the photodimerization of isoprene. Our results suggest a mechanism in which a complex of ferrocene and diene is excited to its second singlet state which dissociates to a triplet-state ferrocene molecule and a triplet-state diene molecule. The triplet-state diene, then, proceeds to isomerize or attack ground-state diene to form dimers.

An investigation of the recaptures rates for tagged brown trout stocked into the River Ribble

The angling season for non-migratory brown trout, in the Environment Agency (EA) North West Region, runs from March 15th to September 30th. Each year, large numbers of farm reared brown trout are stocked into the rivers of the North West Region's Central Area. In 1994, approximately 20,000 brown trout were introduced into the River systems of the Lune, Wyre and Ribble by local angling clubs and fishery owners. Most of these fish were stocked at a length greater than that defined by local byelaws as the takeable size (200mm). Introductions are made to supplement the existing wild brown trout populations within the river and increase the probability of an angler catching a fish. Stocking with fish of a sufficient length allows the successful angler to remove the catch for their own use. In this way, stretches of the rivers are effectively managed as "put and take" fisheries for brown trout. A number of brown trout fingerlings are also introduced each year by angling clubs and fishery owners. These are stocked with the expectation that the fish will survive in the river to grow, over-winter, and eventually attain a takable size with an increased degree of "wildness". The lower cost of fingerlings, as opposed to trout of a takable length, makes their introduction more attractive to angling clubs since a greater number can be stocked for a given cost. Although the practise of stocking brown trout has occurred for many years in the Central Area, there is little information of its success in terms of increasing anglers catches, or the survival offish introduced. This study was initiated to determine the recapture rates by angling of brown trout following their introduction into a river fishery. The information gained from this study can then be used to give guidance to angling clubs and fishery owners on the optimal strategies for stocking fish.