We experimentally detect high-refractive-index media (n > 1.5) using a surface plasmon resonance (SPR) sensor with a diffraction grating. While SPR sensors are generally based on the attenuated total reflection method using metal films, here, we focus on a method using a diffraction grating, which can detect relatively higher refractive-index media and is suitable for device miniaturization. In this study, we used the rigorous coupled-wave analysis method to simulate the dependence of the reflectance on an incident angle for media with refractive index values up to 1.700. In the experiment, a medium (n = 1.660 -1.700) was successfully detected using this grating. Under the conditions of the grating (period: 600 nm, Au thickness: 40 nm) using a red laser (λ: 635 nm), a sharp decline in the reflectance and a rise in the transmittance at certain angles were confirmed, demonstrating the extraordinary transmission enabled by SPR. Because excitation angles changed with changes in the refractive index, we concluded that this method can be applied to sensors that detect high-refractive-index media.
Sperm motility-initiating substance (SMIS) is an oviductal protein critical for internal fertilization in urodeles. It contributes to the establishment of various reproductive modes in amphibians and is thus a unique research model for the gene evolution of gamete-recognizing ligands that have diversified among animal species. In this study, a paralogous SMIS gene, smis2, was identified via the RNA sequencing of the oviduct of the newt, Cynops pyrrhogaster. The base sequence of the smis2 gene was homologous (>90%) to that of the original smis gene (smis1), and deduced amino acid sequences of both genes conserved six cysteine residues essential for the cysteine knot motif. Furthermore, smis2 complementary DNA was identified in the oviduct of Cynops ensicauda, and the base substitution patterns also suggested that the smis gene was duplicated in the Salamandridae. Nonsynonymous/synonymous substitution ratios of smis1 and smis2 genes were 0.79 and 2.6, respectively, suggesting that smis2 gene evolution was independently driven by positive selection. Amino acid substitutions were concentrated in the cysteine knot motif of SMIS2. The smis2 gene was expressed in some organs in addition to the oviduct; in contrast, SMIS1 was only expressed in the oviduct. The SMIS2 protein was suggested to be produced and secreted at least in the oviduct and redundantly act in sperm. These results suggest that smis1 plays the original role in the oviduct, whereas smis2 may undergo neofunctionalization, which rarely occurs in gene evolution.
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