Engineering lysine‐rich caleosins as carrier proteins to render biotin as a hapten on artificial oil bodies for antibody production

It has been demonstrated that caleosin alone is sufficient to stabilize artificial oil bodies. A series of recombinant caleosins, mutated with 3, 5, 8, 11, 13, 15, and 17 extra Lys residues and over‐expressed in Escherichia coli, were used as carrier proteins to render biotin as a hapten on the surface of artificial oil bodies for antibody production. Biotinylation levels of the recombinant caleosins were step‐wisely elevated as the number of extra Lys residues increased, and the biotinylated Lys residues were identified by mass spectrometric analysis. Polyclonal antibodies against biotin were successfully generated in rats injected with artificial oil bodies constituted with each of the biotinylated caleosins. Moreover, those generated via the biotinylated caleosins with eight or more extra Lys residues no longer recognized caleosin. It appears that engineered Lys‐rich caleosins are suitable carrier proteins for the production of antibodies against small molecules. © 2011 American Institute of Chemical Engineers Biotechnol. Prog., 2011     

Reactivities of the coordinated organonitriles in molybdenum(0) and tungsten(0) phosphine complexes: protonation of the nitrile carbon and cleavage of the CN triple bond

The molybdenum and tungsten dinitrogen-organonitrile complexes trans-[M(N₂)(NCR)(dppe)₂] (2, M=Mo; 4, M=W; R=Ph, C₆H₄Me-p, C₆H₄OMe-p, Me; dppe=Ph₂PCH₂CH₂PPh₂) underwent double protonation at the nitrile carbon atom with loss of N₂ and a change in oxidation state to +4 on treatment with hydrochloric acid to afford the cationic imido complexes trans-[MCl(NCH₂R)(dppe)₂]⁺. The solid-state structure of trans-[WCl(NCH₂CH₃)(dppe)₂][PF₆]·CH₂Cl₂ was determined by single-crystal X-ray analysis. Protonation of complexes 2 by fluoroboric acid or hydrobromic acid also formed the similar imido complexes trans-[MoX(NCH₂R)(dppe)₂]⁺ (X=F, Br). In contrast, the dinitrogen complex trans-[Mo(N₂)₂(dppe)₂] reacted with two equiv. of benzoylacetonitrile, a nitrile with acidic CH hydrogen atoms, to give the nitrido complex trans-[Mo(N)(NKCCHCOPh)(dppe)₂] (12), which was accompanied by evolution of dinitrogen and the formation of 1-phenyl-2-propen-1-one in high yields. For complex 12, the zwitterionic structure, where the anionic enolate ligand PhC(O⁺)=CHCN coordinates to the cationic Mo(IV) center through its nitrogen atom, was confirmed by spectroscopic measurements and single-crystal X-ray analysis. A unique intermolecular aromatic C---HO hydrogen bonding was observed in that crystal structure. Complex 12 is considered to be formed via the cleavage of the CN triple bond of benzoylacetonitrile on the metal. A reaction mechanism is proposed, which includes the double protonation of the nitrile carbon atom of the ligating benzoylacetonitrile on a low-valent molybdenum center.     

A study of the occurrence of regulator secondary structures in globular proteins

The distribution of regular secondary structures, viz. α-helices and β-strands, along the length of over 70 properties whose secondary structural details have been reported, has been analysed. The occurrence of these regular structures tends to be a maximum at the N- and C-termini. Our analysis suggests that both these free ends could possibly serve as nucleating centers for secondary structures and could play an important role in the folding of proteins.     

Plant action: Multiple levels of regulation

This lecture is devoted to the relative contribution of various levels of regulation of the actin cytoskeleton functioning in the cell. Regulation at the levels of gene expression, mRNA and protein synthesis and stability, processes of actin polymerization/depolymerization and actin structures reorganization is briefly considered. Novel information about the pathways of signal transduction to the actin cytoskeleton with the involvement of Arp2/3 complex and RIC proteins is highlighted.     

Tree-ring dating of “brush structures” in Kluane National Park and Reserve,Yukon, Canada

“Brush structures” are temporary wooden structures built with unmodified local materials and used as shelters by First Nation Peoples in the forests of the Yukon prior to European contact. This paper reports a preliminary attempt to date these structures using dendrochronology. Investigations were carried out of four njel (“teepee like”) structures and eight män-ku (low 2–3 sided wall structures) at four main sites. The primary material cored was poles (dead spruce trunks), often only 10–20 cm diameter, with narrow, sometimes extremely suppressed ring sequences. These structures are dated between 1865 and 1887, based on the latest (outermost) ring in the sampled material. The limited sampling and use of old wood in these structures (whether fire-kill, standing dead or reused from previous features) makes it difficult to give precise dates for the initial evidence of First Nation activity at these sites: more extensive sampling could provide further insight into the settlement history and construction techniques used. The sites investigated date from the latter half of the nineteenth century shortly before the first European gold rush to this region.     

Using Echo Ultrasound from Schooling Fish to Detect and Classify Fish Types

Fish finders have already been widely available in the fishing market for a number of years.However,the sizes of these fishfinders are too big and their prices are expensive to suit for the research of robotic fish or mini-submarine.The goal of thisresearch is to propose a low-cost fish detector and classifier which suits for underwater robot or submarine as a proximity sensor.With some pre-condition in hardware and algorithms,the experimental results show that the proposed design has good per-formance,with a detection rate of 100 % and a classification rate of 94 %.Both the existing type of fish and the group behaviorcan be revealed by statistical interpretations such as hovering passion and sparse swimming mode.     

The effects of different PMSG doses on estrus behavior and pregnancy rate in Angora goats

Artificial insemination protocols depend on efficient behavioral estrus detection and insemination time in Angora goat. Therefore, we aim to determine the accuracy of an estrus scoring system in Angora goats with different PMSG doses during the breeding season. Does (n: 260) were randomly divided into three groups: group-1 (n: 93), group-2 (n: 85) and group-3 (n: 82). All animals received an intravaginal sponge on day 0 for 11 days, and on the day of sponge insertion 150 μg prostaglandin F2Α was administered. Pregnant mare’s serum gonadotropin was injected 300, 400 and 500 IU intramuscularly 24 h before sponge removal to groups 1, 2 and 3, respectively. Estrus signs were detected with a teaser buck, 24 h after sponge removal according to a visual scoring system. Artificial insemination was performed with 0.25 ml fresh diluted semen at 43 to 45 h after sponge removal. Differences were observed within PMSG groups in terms of standing, tail wagging, courtship behavior, vaginal discharge and vaginal hyperemia (P<0.001). Nevertheless, the most accurate indicators of estrus that result in pregnancy were tail wagging and courtship behavior followed by standing estrus (P<0.05). According to the results obtained, 300 IU PMSG dose is sufficient, both to inseminate at a fixed time (43 to 45 h after sponge removal) and to record the estrus behavior by teaser male 24 h after sponge removal. Higher PMSG doses (400 to 500 IU) altered the timing of ovulation; specifically, 500 IU dose shortened the duration of estrus behaviors. In conclusion, even though the different doses of PMSG displayed similar effects on estrus synchronization and pregnancy rates, we concluded that tail wagging, courtship behavior and standing heat are the most reliable estrus signs for artificial insemination in Angora goat.     

Validation of the first step of the heuristic refinement method for the derivation of solution structures of proteins from NMR data

A new method for the analysis of NMR data in terms of the solution structure of proteins has been developed. The method consists of two steps: first a systematic search of the conformational space to define the region allowed by the initial set of experimental constraints, and second, the narrowing of this region by the introduction of additional constraints and optional refinement procedures. The search of the conformational space is guided by heuristics to make it computationally feasible. The method is therefore called the heuristic refinement method and is coded in an expert system called PROTEAN. The paper describes the validation of the first step of the method using an artificial NMR data set generated from the known crystal structure of sperm whale carbon monoxymyoglobin. It is shown that the initial search procedure yields a low-resolution structure of the myoglobin molecule, accurately reproducing its main topological features, and that the precision of the structure depends on the quality of the initial data set.