Molecular characterization and validation of commercially available methods for haptoglobin measurement in bottlenose dolphin

AbstractHaptoglobin (Hp) is a positive acute-phase protein and a valuable marker of inflammation in both human and veterinary medicine. The aim of this study was to validate the molecular characterization of Hp in dolphins and to validate commercially available Hp measurement methods such as Hp-ELISA (originally designed for pigs) and Hp–hemoglobin (Hb) binding assay. The dolphin Hp (dHp) amino acid sequence appeared most similar to pig Hp by sequence homology and phylogenetic clustering. Amino acid sequence analysis revealed that dHp comprises the Hp1 form of α1 and β chains. The anti-pig Hp antibody cross-reacted with both recombinant dHp, expressed by Escherichia coli, and dHp from serum. The intra- and inter-assay levels of imprecision of pig Hp-ELISA and the Hp–Hb binding assay were found to be tolerable for the determination of Hp in dolphin, and there was no significant discrepancy between the two determination methods. The ability of the assay to differentiate between healthy and inflammation groups was investigated, and a significant increase in Hp concentration was detected in inflammatory conditions. Thus, Hp is a useful inflammation marker for dolphin, and the Hp concentration in dolphin serum samples can be reliably measured using commercially available pig Hp-ELISA and Hp–Hb binding assay

Identification of Personality-Related Candidate Genes in Thoroughbred Racehorses Using a Bioinformatics-Based Approach Involving Functionally Annotated Human Genes

Considering the personality traits of racehorses (e.g., flightiness, anxiety, and affability) is considered essential to improve training efficiency and decrease accident frequency, especially when retraining for a second career that may involve contact with inexperienced personnel after retiring from racing. Studies on human personality-related genes are frequently conducted; however, such studies are rare in horses because a consistent methodology for personality evaluation is lacking. Using the recently published whole genome variant database of 101 Thoroughbred horses, we compared horse genes orthologous to human genes related to the Big Five personality traits, and identified 18 personality-related candidate genes in horses. These genes include 55 variants that involve non-synonymous substitutions that highly impact the encoded protein. Moreover, we evaluated the allele frequencies and functional impact on the proteins in terms of the difference in molecular weights and hydrophobicity levels between reference and altered amino acids. We identified 15 newly discovered genes that may affect equine personality, but their associations with personality are still unclear. Although more studies are required to compare genetic and behavioral information to validate this approach, it may be useful under limited conditions for personality evaluation

Synthesis, Structures, and Properties of π‑Extended Double Helicene: A Combination of Planar and Nonplanar π‑Systems

The synthesis, structures, and properties of a π-extended double helicene <b>1</b> are described. This double helicene <b>1</b> was synthesized by a four-fold oxidative C–H biphenylation of naphthalene followed by the Scholl reaction or via five steps including the Suzuki–Miyaura cross-coupling reaction and the Scholl reaction. Due to the two helical substructures, <b>1</b> has three isomers, i.e., two enantiomers in a twisted form [(<i>P</i>,<i>P</i>) and (<i>M</i>,<i>M</i>)] and one diastereoisomer in a meso form. X-ray crystallographic analysis of the twisted isomers (<i>twisted</i>-<b>1</b>) revealed a tightly offset packing pattern of (<i>P</i>,<i>P</i>)- and (<i>M</i>,<i>M</i>)-twisted isomers, affording a three-dimensional lamellar stacking structure. A high isomerization barrier (43.5 kcal mol^–1) and the relative thermal stability of <i>twisted</i>-<b>1</b> isomer over <i>meso</i>-<b>1</b> by 0.9 kcal mol^–1 were estimated by DFT calculations. The three isomers were successfully separated by chiral HPLC and characterized by circular dichroism spectroscopy as well as by TD-DFT studies. Electronic state variation resulting from the molecular geometry difference between the two diastereoisomers (<i>twisted</i>-<b>1</b> and <i>meso</i>-<b>1</b>) was observed by UV–vis absorption and fluorescence spectra