Step-wise thermal denaturation of cobrotoxin,a snake venom neurotoxin fromNaja naja atra: A proton nuclear magnetic resonance study

Temperature dependence of proton nuclear magnetic resonance spectra has been followed for cobrotoxin, a postsynaptic neurotoxin fromNaja naja atra venom. Several aromatic amino-acid residues, including the functionally essential Trp-29 located at the tip of the central loop of the molecule, have been found to undergo a thermal structural transition above the global thermal denaturation temperature. It is suggested that a local structure around these residues behaves somehow independently of the rest of the molecule, and that such structural organization may be favorable for a conformational change of a neurotoxin molecule on binding to acetylcholine receptor.     

New Attempt at the Synthesis of Lavandulol by a Claisen Type Rearrangement

The use of the Claisen type rearrangement previously studied by K. C. Brannock¹⁾ on some alkyl allyl esters leads in the present study from I, I-dimethyl-2-propenyl senecioate to a mixture of lavandulic acid 70% and iso-lavandulic acid 30%,     

CBP60g and SARD1 play partially redundant critical roles in salicylic acid signaling

Arabidopsis thaliana calmodulin binding protein 60g (CBP60g) contributes to production of salicylic acid (SA) in response to recognition of microbe‐associated molecular patterns (MAMPs) such as flg22, a fragment of bacterial flagellin. Calmodulin binding is required for the function of CBP60g in limiting growth of the bacterial pathogen Pseudomonas syringae pv. maculicola (Pma) ES4326 and activation of SA synthesis. Here, we describe a closely related protein, SARD1. Unlike CBP60g, SARD1 does not bind calmodulin. Growth of Pma ES4326 is enhanced in sard1 mutants. In cbp60g sard1 double mutants, growth of Pma ES4326 is greatly enhanced, and SA levels and expression of PR‐1 and SID2 are dramatically reduced. Expression profiling placed the CBP60g/SARD1 node between the PAD4/EDS1 and SA nodes in the defense signaling network, and indicated that CBP60g and SARD1 affect defense responses in addition to SA production. A DNA motif bound by CBP60g and SARD1, GAAATTT, was significantly over‐represented in promoters of CBP60g/SARD1‐dependent genes, suggesting that expression of these genes is modulated by CBP60g/SARD1 binding. Gene expression patterns showed a stronger effect of cbp60g mutations soon after activation of a defense response, and a stronger effect of sard1 mutations at later times. The results are consistent with a model in which CBP60g and SARD1 comprise a partially redundant protein pair that is required for activation of SA production as well as other defense responses, with CBP60g playing a more important role early during the defense response, and SARD1 to playing a more important role later.     

Studies on Phenolic Lactones

Isohibalactone, the geometric isomer of hibalactone, was synthesized by the following route. Piperonylsuccinic acid anhydride was converted into thioethyl methyl ester ans was reduced to piperonylbutyrolactone by Raney nickel catalyst. Piperonylbutyrolactone was also prepared from piperonylsuccinic anhydride by the reduction with amalgamated aluminum. Condensation of piperdnal with the lactone in the presence of potassium amide afforded α-(3,4-methyIenedioxy-phenyl-hydroxymethyl)-β-(3,4-methylenedioxybenzyl)-butyrolactone, m.p. 151~2°C. Dehydration of the hydroxylactone with p-toluenesulfonic acid gave isohibalactone, m.p. 156~6.5°C.     

Studies on Phenolic Lactones

The synergistic action of phenolic lactones on allethrin and pyrethrin were investigated from the mortality of synergized pyrethroids against rice-weevil by petri-dish method.

In the series of α-benzylidene-γ-butyrolactones, (±) hibalactone and α-piperonylidenebutyrolactone were appreciably sy~ergistic on allethrin although less effective than piperonyl butoxide. (±) Hinokinin, 2-piperonylidene-3-piperonyl-l,4-butanediol also showed week activation, but α-benzylidene-, α-anisylidene-, α-veratrylidene-butyrolactone, α-piperonylidenea-α’-piperonyl-tetrahydrofuran, α-trirnethoxybenzylidene-β-trimethoxybenzyl-butyrolactone were not synergistic on the insecticidal action.

In the series of synergized pyrethrin, (±) hibalactone and α-piperonylidene-butyrolactone showed week synergism but the other test compounds showed no appreciable synergism.     

Synthetic Studies on Colchicine-related Tropolones

Pschorr reaction of 4-[3-(3,4,5-trimethoxyphenyl)-propyl]-5-amino-tropolone (VII) gave desacetamidocolchiceine (IIa), though the oxidative coupling reaction of 4-[3-(3,4-dihydroxy- 5-methoxy phenyl)-propyl]-tropolone (Xa) failed to give the same compound (IIa).     

Sex-biased natal dispersal in Hokkaido brown bears revealed through mitochondrial DNA analysis

Understanding natal dispersal patterns is fundamental in the ecology and conservation biology of large wild carnivores. In this study, we used two approaches to determine genetic variation and dispersal patterns of brown bears in the Shiretoko Peninsula, eastern Hokkaido, Japan. The first approach was a large-scale genetic analysis. We analyzed haplotypes from the mitochondrial DNA (mtDNA) control region of 760 individual samples collected throughout the peninsula during 1998–2016. We detected seven haplotypes, including two that were confirmed for the first time. In females, the distribution of haplotypes was geographically structured, whereas haplotypes in males were distributed widely throughout the peninsula. Only some males in the lower peninsula had haplotypes that were not detected within the peninsula. The second approach was a local-scale genetic analysis, including intensive focal sampling in the Rusha area, a special wildlife protection area on the peninsula. Proportions of mtDNA haplotypes in adult bears were investigated and compared between the sexes. Although more than half of the females had the same haplotype, males had more diverse haplotypes, suggesting that they came to the Rusha area from other regions. Thus, our study revealed that mtDNA haplotype distribution has been maintained by female philopatry, and that bears exhibit male-biased dispersal. Furthermore, the lower peninsula appears to act as a contact zone between the peninsula and mainland Hokkaido, which is important for maintaining genetic diversity.