アルツハイマー型認知症者を対象にした机上で実施可能なIADL検査の開発と妥当性の検討

The instrumental activities of daily living scale (IADL scale) for Alzheimer’s type of dementia (ATD) does not provide a simple and easy way for evaluators to directly evaluate the targeted individuals. Therefore, we developed an experimental method, which allows a desk evaluation of IADL (on-the-desk evaluation of IADL). The study involved a group of 24 subjects with normal control (MMSE, 28.9 ± 1.4) and a group of 21 subjects with ATD (MMSE, 19.8 ± 4.4). The desk evaluation of IADL involved 9 tasks, which included boiling water with an electric kettle, pulling the plug out of the outlet once the water boils, and making a telephone call. For the test, the subjects were instructed to figure out by themselves the sequence in which they would perform the tasks. The scoring was performed on the basis of the appropriateness or inappropriateness of their behavior in each task and the time it took them to complete each task. The maximum score was set to a total of 59 points. Additionally, in order to examine the validity of the test and the factors which may affect the desk assessment of IADL, various types of neuropsychological tests were performed. The IADL Scale and the Frenchay Activities Index (FAI) were used in the evaluation of IADL. The results showed significant difference in the scores between the 2 groups. The at-the-desk evaluation of IADL revealed a strong correlation between the IADL Scale and the FAI (r＝0.89, r＝0.82, p＜0.001), and a multiple regression analysis of the IADL scores from the desk evaluation showed a high explanation rate (R2＝0.84) by “the Behavioural Assessment of the Dysexecutive Syndrome, “the Wechsler Memory Scale-Revised/digit span backwards test,” the “Rivermead Behavioural Memory Test/appointment”, “Composing task”. The high correlation between the desk evaluation of IADL and the IADL evaluation scale and the results of the multiple regression analysis indicated that the desk evaluation of IADL reflected cognitive functions in ATD and was highly reliable and valid. アルツハイマー型認知症（ATD）用の手段的日常生活活動（IADL）スケールは、評価 者が直接対象者に対して簡便に評価可能なものはない。そこで、我々は机上で実施可能な IADL評価方法（机上IADL検査）の開発を試みた。対象は、健常群２４名（MMSE２８９. ±１４. ） とATD群２１名（MMSE１９８. ±４４. ）とした。机上IADL検査の課題は、電気ポットでお湯を 沸かす、お湯が沸いたらコンセントを抜く、電話をかける、などのIADLの課題を９つ設定 した。また、検査規則として、課題実施順番は自分で考えること、などを伝えた。得点化 は、各課題の適切・不適切行動と検査実施時間、実施順序を対象に行い、合計５９点満点と した。さらに、机上IADL検査に影響する因子と妥当性を検討するために、各神経心理学的 検査を実施し、IADL評価スケールとして、IADL ScaleとFenchay Activities Index（FAI） を実施した。その結果、２群の得点比較では有意差が認められた。また、机上IADL検査 は IADL Scale、FAI ともに強い相関を認め（r =０ ８.９、r =０ ８.２、p＜００.０１）、重回帰分析で は、机上IADL検査得点に対し「遂行機能症候群の行動評価」、「改訂ウェクスラー記憶検 査・逆唱課題」「リバミード行動記憶検査・約束課題」「構成課題」により、高い寄与率 （R２＝０８.４）が示された。机上 IADL 検査は、IADL 評価スケールとの相関が高いこと、重 回帰分析の結果から、ATDの認知機能をよく反映した併存妥当性の高い検査と考えられた

細菌の接種によって誘導されるアコヤガイ中の抗菌性タンパク質の探索

The aim of this research was to characterize immune-related antibacterial substances from pearl oyster Pinctada fucata induced by bacterial invasion. Bacteria inoculation was performed by injecting 0.1 ml of 1.0 × 1012 colony-forming units/ml Vibrio parahaemolyticus into adductor muscle. Acidic extracts were prepared with 0.1% trifluoroacetic acid from different tissues after 8 h of injection, and antibacterial activity against V. parahaemolyticus was determined via the microdilution broth method. The acidic extracts from gills of inoculated oysters (AEg) showed stronger antibacterial activity than those from non-inoculated ones. Based on this result, antibacterial proteins were purified from AEg via two-step gel filtration chromatography, followed by high-performance liquid chromatography using a TSkgel G3000 column. Protein components were analyzed by both sodium dodecyl sulfate and native polyacrylamide gel electrophoresis. As a result, two antibacterial proteins, APg-1 (with a molecular mass of approximately 210 kDa) and APg-2 (of approximately 30 kDa), were obtained from AEg. Matrix-assisted laser desorption/ionization time of flight mass spectrometry analysis and partial amino acid sequences revealed that these proteins might be novel antibacterial proteins. These results indicate that antibacterial proteins are potentially upregulated in the gill of pearl oysters or released therefrom for defense against bacterial invasion.本研究で対象とするアコヤガイは，日本において重要な養殖真珠の母貝として用いられる二枚貝である。腸炎ビブリオをアコヤガイ閉殻筋に直接接種したところ，鰓から得られた酸抽出物に，非接種の対照よりも強い抗菌活性を示す成分が存在することを見出した。酸抽出物はグラム陽性菌および陰性菌に抗菌活性を示し，とくにビブリオ属に強く作用した。鰓より2種の抗菌タンパク質APg-1（分子量約210 kDa）およびAPg-2（分子量約30 kDa）を分離し，これらはMALDI-TOF MS分析により，新規の抗菌タンパク質である可能性が推察された

アルツハイマー病者のIADL に影響する高次脳機能障害についての検討 : 机上IADL 検査を用いて

はじめに：アルツハイマー病（AD）の軽度段階では、いくつかの手段的日常生活活動（IADL） の能力が低下し、高次脳機能の低下やうつがIADL に影響していると考えられている。し かし、IADL と高次脳機能、うつとの関連について検討した報告は少ない。そこで、我々 は「机上IADL 検査」を用いて、高次脳機能やうつがIADL にどのように影響しているの かについて検討した。方法：対象は軽度AD 群25 名と健常高齢者群22 名とし、7 種類の IADL 課題で構成され信頼性と妥当性が証明されている、「机上IADL 検査」と神経心理 学的検査、簡易うつ検査（GDS-15）を実施した。結果：軽度AD 群では、「服をしまう」、「ご みの分別」、「本の分別」の課題については、実施割合が高い結果となった。しかし、すべ てのIADL 課題で誤反応が健常高齢者より有意に多い結果となった。7 種類のIADL 課題 と神経心理学的検査とGDS-15、年齢との関連を検討したところ、神経心理学的検査によっ て相関の強さの違いがみられた。また各IADL 課題は、短期記憶との関連が強かった。 考察：軽度AD 者は、誤り行動を含みながらIADL タスクを実施していると考えられた。 また、軽度AD 者へのIADL 支援を行う際は、影響する認知機能の種類に配慮した検査の 選択が必要と考えられた

A high-quality, haplotype-phased genome reconstruction reveals unexpected haplotype diversity in a pearl oyster

Homologous chromosomes in the diploid genome are thought to contain equivalent genetic information, but this common concept has not been fully verified in animal genomes with high heterozygosity. Here we report a near-complete, haplotype-phased, genome assembly of the pearl oyster, Pinctada fucata, using hi-fidelity (HiFi) long reads and chromosome conformation capture data. This assembly includes 14 pairs of long scaffolds (>38 Mb) corresponding to chromosomes (2n = 28). The accuracy of the assembly, as measured by an analysis of k-mers, is estimated to be 99.99997%. Moreover, the haplotypes contain 95.2% and 95.9%, respectively, complete and single-copy BUSCO genes, demonstrating the high quality of the assembly. Transposons comprise 53.3% of the assembly and are a major contributor to structural variations. Despite overall collinearity between haplotypes, one of the chromosomal scaffolds contains megabase-scale non-syntenic regions, which necessarily have never been detected and resolved in conventional haplotype-merged assemblies. These regions encode expanded gene families of NACHT, DZIP3/hRUL138-like HEPN, and immunoglobulin domains, multiplying the immunity gene repertoire, which we hypothesize is important for the innate immune capability of pearl oysters. The pearl oyster genome provides insight into remarkable haplotype diversity in animals

Phylogenetic comparisons reveal mosaic histories of larval and adult shell matrix protein deployment in pteriomorph bivalves

Molluscan shells are organo-mineral composites, in which the dominant calcium carbonate is intimately associated with an organic matrix comprised mainly of proteins and polysaccharides. However, whether the various shell matrix proteins (SMPs) date to the origin of hard skeletons in the Cambrian, or whether they represent later deployment through adaptive evolution, is still debated. In order to address this issue and to better understand the origins and evolution of biomineralization, phylogenetic analyses have been performed on the three SMP families, Von Willebrand factor type A (VWA) and chitin-binding domain-containing protein (VWA-CB dcp), chitobiase, and carbonic anhydrase (CA), which exist in both larval and adult shell proteomes in the bivalves, Crassostrea gigas and Pinctada fucata. In VWA-CB dcp and chitobiase, paralogs for larval and adult SMPs evolved before the divergence of these species. CA-SMPs have been taken as evidence for ancient origins of SMPs by their presumed indispensable function in biomineralization and ubiquitous distribution in molluscs. However, our results indicate gene duplications that gave rise to separate deployments as larval and adult CA-SMPs occurred independently in each lineage after their divergence, which is considerably more recent than hitherto assumed, supporting the "recent heritage and fast evolution" scenario for SMP evolution

Dual Gene Repertoires for Larval and Adult Shells Reveal Molecules Essential for Molluscan Shell Formation

Molluscan shells, mainly composed of calcium carbonate, also contain organic components such as proteins and polysaccharides. Shell organic matrices construct frameworks of shell structures and regulate crystallization processes during shell formation. To date, a number of shell matrix proteins (SMPs) have been identified, and their functions in shell formation have been studied. However, previous studies focused only on SMPs extracted from adult shells, secreted after metamorphosis. Using proteomic analyses combined with genomic and transcriptomic analyses, we have identified 31 SMPs from larval shells of the pearl oyster, Pinctada fucata, and 111 from the Pacific oyster, Crassostrea gigas. Larval SMPs are almost entirely different from those of adults in both species. RNA-seq data also confirm that gene expression profiles for larval and adult shell formation are nearly completely different. Therefore, bivalves have two repertoires of SMP genes to construct larval and adult shells. Despite considerable differences in larval and adult SMPs, some functional domains are shared by both SMP repertoires. Conserved domains include von Willebrand factor type A (VWA), chitin-binding (CB), carbonic anhydrase (CA), and acidic domains. These conserved domains are thought to play crucial roles in shell formation. Furthermore, a comprehensive survey of animal genomes revealed that the CA and VWA-CB domain-containing protein families expanded in molluscs after their separation from other Lophotrochozoan linages such as the Brachiopoda. After gene expansion, some family members were co-opted for molluscan SMPs that may have triggered to develop mineralized shells from ancestral, nonmineralized chitinous exoskeletons

Electroextraction of Insoluble Proteins from the Organic Matrix of the Nacreous Layer of the Japanese Pearl Oyster, <i>Pinctada fucata</i>

The nacreous layer of shells and pearls is composed of aragonite crystals arranged in an organic matrix. The organic matrix contains chitin and several proteins that regulate the formation of the nacreous layer. Owing to their strong interactions in the organic matrix, the current method for extraction of insoluble proteins from the pre-powdered nacreous layer involves heating to high temperatures in the presence of a detergent (e.g., sodium dodecyl sulfate, SDS) and reductant (e.g., dithiothreitol, DTT), which is likely to induce protein degradation. Therefore, we have developed an electroextraction method to isolate proteins from the organic matrix of a nacreous organic sheet, that was obtained following the decalcification of shells in their original shape. Our electroextraction method employs milder conditions without heating or detergent. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) patterns of the electro-extracted proteins (EEPs) under non-reduced and reduced conditions revealed that this method yielded a greater number of different proteins compared with the conventional extraction method and the isolated EEPs retained their disulfide bonds. Our method is able to easily extract insoluble proteins from the nacreous layer under mild conditions and will undoubtedly aid future analyses into the functions of the nacreous layer proteins