Please press and hold the button until it turns completely green If you believe this is an error, please contact our support team. 147.45.197.102 : 94f46542-1bfb-4ccb-9531-a839eae4 Volume 16 - 2022 | https://doi.org/10.3389/fnins.2022.807473 This article is part of the Research TopicAdvances in the Imaging Techniques of Radiologically subtle CNS DisordersView all 13 articles more than 100 transgenic (Tg) mouse models of Alzheimer’s disease (AD) have been generated in which mutant amyloid precursor protein (APP) or APP/presenilin 1 (PS1) cDNA is overexpressed (1st generation models) Although many of these models successfully recapitulate major pathological hallmarks of the disease such as amyloid β peptide (Aβ) deposition and neuroinflammation they have suffered from artificial phenotypes in the form of overproduced or mislocalized APP/PS1 and their functional fragments as well as calpastatin deficiency-induced early lethality Such artifacts bring two important uncertainties into play and (2) how they affect the interpretation of experimental results destruction of endogenous gene loci in some Tg lines by transgenes has been reported single App knock-in mouse models harboring the Swedish and Beyreuther/Iberian mutations with or without the Arctic mutation (AppNL–G–F and AppNL–F mice) were developed (2nd generation models) While these models are interesting given that they exhibit Aβ pathology and cognitive impairment in an age-dependent manner which exhibits an extensive pathology as early as 6 months of age is not suitable for investigating Aβ metabolism and clearance because the Aβ in this model is resistant to proteolytic degradation and is therefore prone to aggregation it cannot be used for preclinical immunotherapy studies owing to the discrete affinity it shows for anti-Aβ antibodies The weakness of the latter model (without the Arctic mutation) is that the pathology may require up to 18 months before it becomes sufficiently apparent for experimental investigation this model was successfully applied to modulating Aβ pathology by genome editing to revealing the differential roles of neprilysin and insulin-degrading enzyme in Aβ metabolism and to identifying somatostatin receptor subtypes involved in Aβ degradation by neprilysin we also provide here a technical guide for the application of App knock-in mice to AD research a new double knock-in line carrying the AppNL–F and Psen1P117L/WT mutations was generated the pathogenic effect of which was found to be synergistic A characteristic of this 3rd generation model is that it exhibits more cored plaque pathology and neuroinflammation than the AppNL–G–F line and thus is more suitable for preclinical studies of disease-modifying medications targeting Aβ a derivative AppG–F line devoid of Swedish mutations which can be utilized for preclinical studies of β-secretase modifier(s) was recently created we introduce a new model of cerebral amyloid angiopathy that may be useful for analyzing amyloid-related imaging abnormalities that can be caused by anti-Aβ immunotherapy Use of the App knock-in mice also led to identification of the α-endosulfine-KATP channel pathway as components of the somatostatin-evoked physiological mechanisms that reduce Aβ deposition via the activation of neprilysin Such advances have provided new insights for the prevention and treatment of preclinical AD Because tau pathology plays an essential role in AD pathogenesis knock-in mice with human tau wherein the entire murine Mapt gene has been humanized were generated the carboxy-terminal PDZ ligand of neuronal nitric oxide synthase (CAPON) was discovered as a mediator linking tau pathology to neurodegeneration and showed that tau humanization promoted pathological tau propagation we describe and discuss the current status of mutant human tau knock-in mice and a non-human primate model of AD that we have successfully created Mislocalization of APP in APP-overexpressing mice APP23 (APP-overexpressing mice) and App KI mice (AppNL–F/NL–F) were subjected to immunohistochemistry using antibodies to APP a synaptic vesicle marker (lower panels) as indicated App KO mice were used as negative controls for APP staining While APP is selectively expressed in the axons of WT and KI mice APP23 expresses unphysiologically high levels of APP not only in the axons but also in the soma and dendrites indicating that the mutant mice are relevant models for studying AD in general Second generation mouse models of Alzheimer’s disease Successful application of the 2nd generation mouse models AppG–F mice suitable for studies of BACE1 inhibitors The AppG–F line is devoid of the Swedish mutation that influences the β-secretase activity and elevates the quantity of CTFβ (The AppG–F line instead carries a wild-type sequence: KM.) The AppG–F model would be appropriate for use in preclinical studies of β-secretase inhibitors without the interference of the Swedish mutation List of mutant mice that are and will be made available to the research community which is too long for researchers to wait in a practical sense a new mouse model that accumulates wild-type human Aβ as quickly as the AppNL–G–F model but did not depend on the presence of the Arctic mutation was desired Scheme of AppNL–F × Psen1P117L double-mutant mice For the generation of the double-mutant mice the AppNL–F line was crossbred with the Psen1P117L line whose mutation was introduced in the endogenous Psen1 gene utilizing base editing technology The synergistic effects of the pathogenic mutations in the App and Psen1 genes strongly accelerates the deposition of wild-type human Aβ in mouse brains AD pathology in the hippocampus of a 3rd generation model mouse A 12-month-old AppNL–F X Psen1P117L/WT mouse was analyzed by immunohistochemistry Blue: Aβ plaques; red: microglia; green: astrocytes Characteristics of the App knock-in mouse lines There are several precautions to be aware of to make the best use of the App knock-in mice A number of the App knock-in mouse users use incorrect nomenclature such as APP-NLF APPNLF and APPNL–F instead of the AppNL–F mice which accords with international rules of standard genomic nomenclature Genetic names always need to be italicized The method allows the most sensitive quantification of both soluble and insoluble Aβ with the smallest protocol deviations Outlined protocols for extraction and quantification of Aβ from tissues there is no restriction regarding use of antibodies for the AppNL–F X Psen1P117L mice Reactivity of different antibodies to Arctic Aβ in AppNL–G–F mice (A) Epitope map of anti-Aβ antibodies (B,C) Quantification of Arctic Aβ species using BNT77 as a capture antibody BNT77 binds to the mid-portion of Aβ [see epitope map (A)] Japan) was used to quantify Aβx-40 (C) and Aβx-42 (D) (D,E) Quantification of Arctic Aβ species using BAN50 as a capture antibody BAN50 binds to the N-terminal region of Aβ [see epitope map (A)] Japan) was used to quantify Aβx-40 (D) and Aβx-42 (E) BNT77 and BAN50 captured Arctic Aβ more weakly than wild-type Aβ (F) Immunohistochemistry using various anti-Aβ antibodies Brain sections derived from AppNL–F mice (24 months old) were immunostained using antibodies with different epitopes after antigen retrieval as indicated (upper panels); those of AppNL–G–F mice (9 months old) were similarly immunostained (lower panels) it’s possible that low level of endothelial APP expression in mice could be one of the reasons that App knock-in mice exhibit mild CAA pathology a mouse line that specifically expresses human APP770 in endothelial cells has just been generated (unpublished) floxed hAPP770NL mice under the CMV early enhancer/chicken β-actin promoter were first generated These mice were then crossed with Tie2-Cre mice in which the Tie2 promoter directs the expression of Cre recombinase in the endothelial cells to obtain double transgenic (Tg) mice App knock-in mouse models were previously produced by Li et al. (2014) who used multiple pathogenic mutations. These mice carry the Swedish (K670N/M671L), Dutch (E693Q), and London (V717I) mutations with the humanized Aβ sequence. The Dutch mutation results in an intensive CAA pathology in humans, thereby causing brain hemorrhage and early mortality (Levy et al., 1990; Van Broeckhoven et al., 1990) This mutation is therefore not specifically responsible for causing FAD These mice did not develop prominent Aβ deposits over their lifespan but when they were crossbred with Psen1M146V knock-in mice an age-dependent deposition of Aβ was seen in the resultant double knock-in mice The deposition of Aβ was detected not only in the parenchyma of the cerebral cortex but also in the cerebral vasculature Double knock-in mice that did not have the Dutch mutation exhibited virtually no vascular pathology if the authors had used the Beyreuther/Iberian or Austrian mutation instead of the London mutation in the mouse App gene then they probably would not have had to introduce the Psen1 knock-in mice Knock-in mice harboring the Dutch mutation can still serve as relevant models for CAA; however they may not be appropriate for examining the effect of immunotherapy on CAA because the Dutch mutation is present in the middle of the Aβ sequence rather IDE appears to be involved in metabolism of AICD their roles in Aβ metabolism in vivo have never been compared in an impartial and side-by-side manner Once double mutants crossbred single App knock-in mice with NEP (Mme) KO mice and with IDE (Ide) KO mice were analyzed in detail for their biochemical properties and Aβ pathology properties it would be clear their distinct roles in APP metabolism and the AD pathogenesis our results imply that the aging-associated decrease in NEP expression is a primary cause of SAD and could thus be a target for the treatment of preclinical AD once other factors such as apolipoprotein E genotypes have also been considered Somatostatin receptor subtypes 1 and 4 (SST1/4) regulate the Aβ-degrading enzyme NEP The neuropeptide somatostatin (SRIF) was identified as a regulator of NEP activity through in vitro screening Subsequent analysis of the effect of genetic deletion of somatostatin receptor (SST) subtypes in mice revealed that SST1 and SST4 regulate NEP in a redundant manner This was further confirmed by concurrently deleting SST1 and SST4 in App KI mice SST1/4 can be either a combination of SST1 and SST4 homodimers or an SST1/SST4 heterodimer Although SRIF is known to regulate Aβ catabolism by enhancing NEP-catalyzed proteolytic degradation, the mechanism by which SRIF actually regulates NEP activity is yet to be fully elucidated. Proteomic analyses enabled α-endosulfine (ENSA), an endogenous ligand of the ATP-sensitive potassium (KATP) channel, to be identified as a negative regulator of NEP downstream of SRIF signaling (Watamura et al., 2021a) The expression of ENSA is significantly increased in AD mouse models and in patients with AD NEP directly contributes to the degradation of ENSA suggesting a substrate-dependent feedback loop regulating NEP activity Role of ENSA in the regulation of NEP activity Schematic illustration of the mechanism describing NEP activity in the brain a downstream protein of SST-SST1/4 signaling plays a role as a ligand of the KATP channel composed of sulfonylurea receptor subunit 1 (SUR1) and inwardly rectifying K+ channel 6.2 (Kir6.2) To date, most if not all, mouse models of tauopathy have been unable to recapitulate the tau pathology without overexpressing mutant human tau protein. As a novel in vivo platform for studying human tauopathy, human MAPT knock-in mice have been developed in which the entire Mapt gene including all exons and introns are humanized (Hashimoto et al., 2019) the MAPT and Mapt genes encoded human and murine tau proteins This was done by crossing MAPT knock-in mice with single App knock-in mice in order to study the role of the Aβ-tau axis in the etiology of AD The double knock-in mice exhibited a more pronounced tau phosphorylation status than single MAPT knock-in mice but lacked evidence of tau pathology and neurodegeneration (in a manner similar to that of single App knock-in mice) even after waiting until the mice were 24 months old These mice exhibit aging-dependent tau aggregation and cognitive impairment in a manner accelerated by Aβ pathology and are being provided to the research community upon request Propagation of tau in each mouse line was observed 3 months after AD-tau injection Brain sections were immunostained with AT8 (red) Humanization of the host animal tau affects the transmission of the pathogenic agents AppNL–G–F/MAPT dKI mice exhibited greater pathological propagation than AppNL–G–F KI mice We also previously identified a NMDAR-related molecule as a tau binding protein which is involved in tau pathology- induced neurodegeneration To elucidate key molecules underlying tau accumulation-induced neurodegeneration, a comprehensive screening of tau-interacting proteins (tau interactome) was constructed. Tau-binding proteins were isolated by immunoprecipitation-LC-MS/MS (IP-MS) using a Flag-tag antibody and wild-type tau Tg (wtau-Tg) mice, which is expressing human tau tagged with a flag epitope (Kimura et al., 2007) Considering that tau is a microtubule-binding protein we validated the methods used to generate the tau interactome by identifying the tubulin beta-4A chain as one of the tau-binding proteins implying that CAPON may play an important role in the pathogenesis of AD The mechanism(s) underlying these effects nevertheless remain(s) unknown an intervention in the interaction between CAPON-tau or CAPON-nNOS could be a new approach for the treatment of AD P301S-Tau-Tg or nos1ap-/-/P301S-Tau-Tg mice stained by the conventional method using hematoxylin and eosin (H&E) A CAPON (Nos1ap) deficiency restores AD-related pathological phenotypes in P301S-Tau-Tg mice Aβ pathology elevates the level and localization of CAPON in hippocampal pyramidal cells CAPON-induced neuronal cell death is closely associated with the pathological tau protein although there appears to be a tau-independent mechanism in play as well Photograph of common marmosets (Callithrix jacchus) The photo shows members of captive common marmoset family and high cognitive functions are a suitable model for neuroscience The photograph of marmosets was taken by WK at CIEA All authors listed have made a substantial Part of the work described was supported by the following research grants: JSPS KAKENHI Grant Number JP18K07402 (HS); JSPS KAKENHI Grant Numbers JP15K19036 and JP19K16271 and RIKEN Special Postdoctoral Research program (SH); JSPS KAKENHI Grant Number JP21K15378 (NW); AMED Grant Number JP19dm0207065 (ES) AMED Grant Number JP18am0101036 (SK); The Swedish Research Council 2018-02843 (BW); Hållsten Research Foundation (PN); JSPS KAKENHI Grant Number JP20H03564/AMED Grant Numbers JP21gm1210010s0102 and JP21dk0207050h001/JST [Moonshot R&D] Grant Number JPMJMS2024/Grant-in-aid for Research in Nagoya 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This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY) distribution or reproduction in other forums is permitted provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited in accordance with accepted academic practice distribution or reproduction is permitted which does not comply with these terms *Correspondence: Takashi Saito, c2FpdG8tdEBtZWQubmFnb3lhLWN1LmFjLmpw; Nobuhisa Iwata, aXdhdGEtbkBuYWdhc2FraS11LmFjLmpw; Takaomi C. Saido, dGFrYW9taS5zYWlkb0ByaWtlbi5qcA== Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher 94% of researchers rate our articles as excellent or goodLearn more about the work of our research integrity team to safeguard the quality of each article we publish Metrics details Base Editor (BE) and Target-AID (activation-induced cytidine deaminase) are engineered genome-editing proteins composed of Cas9 and cytidine deaminases These base-editing tools convert C:G base pairs to T:A at target sites we inject either BE or Target-AID mRNA together with identical single-guide RNAs (sgRNAs) into mouse zygotes and compare the base-editing efficiencies of the two distinct tools in vivo BE consistently show higher base-editing efficiency (10.0–62.8%) compared to that of Target-AID (3.4–29.8%) unexpected base substitutions and insertion/deletion formations are also more frequently observed in BE-injected mice or zygotes We are able to generate multiple mouse lines harboring point mutations in the mouse presenilin 1 (Psen1) gene by injection of BE or Target-AID These results demonstrate that BE and Target-AID are highly useful tools to generate mice harboring pathogenic point mutations and to analyze the functional consequences of the mutations in vivo researchers need to design template DNA for each different mutation it remains unclear whether it works in mammals Here we introduced either BE or Target-AID mRNA together with identical sgRNAs that targeted the Psen1 or App gene into mouse zygotes Multiple animal models were generated with a number of distinct disease-related and disease-unrelated point mutations thereby enabling the functional consequences of these mutations to be evaluated in vivo Generation of Psen1-P436S mice by BE and Target-AID The activity windows of BE (pink) and Target-AID (light blue) are indicated b Schematic representation of the injection of mouse zygotes for the generation of Psen1-P436S mice BE or Target-AID mRNA was injected together with sgRNA-Psen1-P436 into the cytoplasm of mouse zygotes to generate mutant Psen1 mice d Sanger sequencing chromatograms in BE-injected (c) and Target-AID-injected (d) mice Magenta arrowheads indicate expected substitutions while green arrowheads indicate undesired substitutions The panels on the right represent Sanger sequencing chromatograms of F1 mice generated from the #89 (BE-injected F0) mouse (c) or the #156 (Target-AID-injected F0) mouse (d) The column on the right indicates the frequency of each genotype in the F1 generation and these mutations could have influenced our results These results indicate that BE has a high base-editing efficiency but with less precision whereas Target-AID edits bases less efficiently but with relatively high accuracy Base-editing tools should thus facilitate exploration of the unidentified functional consequences of each pathogenic mutation It is therefore possible that base excision repair initiated by uracil DNA glycosirase may be responsible for causing DSB and subsequent indel formation which might explain the low base-editing efficiency in mouse embryos in the present study Although the base-editing efficiency of Target-AID was lower than that of BE in mice the frequency of undesired conversion and indel formation was also lower In addition to targeting the 2nd and 3rd Cs in sgRNA sequences Target-AID might also be a better option than BE for introducing precise point mutations targeting the 4th C Screening using base-editing technology could therefore prove valuable for the identification of such unknown pathogenic mutations or protective variants The sgRNAs were synthesized using the MEGAshortscript T7 Transcription Kit (Thermo Fisher Scientific Transcribed mRNAs and sgRNAs were purified using the MEGAclear Transcription Clean-Up Kit (Thermo Fisher Scientific and 30 ng/μl for Psen1-P117) were injected into the cytoplasm of C57BL/6J zygotes embryos at the 2-cell-stage were transferred to host ICR female mice The targeted deep sequencing was performed using an Illumina Miseq system Immunoreactive bands on the membrane were visualized with ECL Select (GE Healthcare) and scanned with a LAS-3000mini Lumino Image analyzer (Fujifilm) Immunohistochemical staining against Aβ (N1D 1:200) was performed in 9-month-old Psen1-P436S mice and 24-month-old NL-F mice After deparaffinization of paraffin-embedded mouse brain sections antigen retrieval was performed by autoclave treatment (121 °C for 5 min) and then incubated with primary antibody in blocking solution overnight A biotinylated secondary antibody and tyramide signal amplification (PerkinElmer) were used for detection of amyloid pathology Photographic data were captured using a NanoZoomer Digital Pathology C9600 (Hamamatsu Photonics) we used one-way analysis of variance followed by Tukey’s post hoc analysis The data were collected and processed in a randomized and blinded manner Sequencing information has been uploaded to DNA Data Bank of Japan (DDBJ) Sequencing Read Archive (DRA) with the accession code DRA007028 The data related to the findings of this study are available from the corresponding authors on request Guidelines for investigating causality of sequence variants in human disease Of mice and CRISPR: the post-CRISPR future of the mouse as a model system for the human condition CRISPR-based technologies for the manipulation of eukaryotic genomes Targeted nucleotide editing using hybrid prokaryotic and vertebrate adaptive immune systems Increasing the genome-targeting scope and precision of base editing with engineered Cas9-cytidine deaminase fusions Highly efficient RNA-guided base editing in mouse embryos Improving the DNA specificity and applicability of base editing through protein engineering and protein delivery Programmable base editing of zebrafish genome using a modified CRISPR-Cas9 system Highly efficient and precise base editing in discarded human tripronuclear embryos Highly efficient base editing in human tripronuclear zygotes Targeted base editing in rice and tomato using a CRISPR-Cas9 cytidine deaminase fusion Pathogenic presenilin 1 mutations (P436S & I143F) in early-onset Alzheimer’s disease in the UK Alzheimer’s disease genetics: from the bench to the clinic Cas-OFFinder: a fast and versatile algorithm that searches for potential off-target sites of Cas9 RNA-guided endonucleases COSMID: a web-based tool for identifying and validating CRISPR/Cas off-target sites Photoactivated gamma-secretase inhibitors directed to the active site covalently label presenilin 1 Increased vulnerability of hippocampal neurons to excitotoxic necrosis in presenilin-1 mutant knock-in mice Accumulation of murine amyloidbeta42 in a gene-dosage-dependent manner in PS1 ‘knock-in’ mice Presenilin-1 P264L knock-in mutation: differential effects on abeta production Massive CA1/2 neuronal loss with intraneuronal and N-terminal truncated Abeta42 accumulation in a novel Alzheimer transgenic model Enzymatic characteristics of I213T mutant presenilin-1/gamma-secretase in cell models and knock-in mouse brains: familial Alzheimer disease-linked mutation impairs gamma-site cleavage of amyloid precursor protein C-terminal fragment beta Potent amyloidogenicity and pathogenicity of Abeta43 Presenilin-1 knockin mice reveal loss-of-function mechanism for familial Alzheimer’s disease The Psen1-L166P-knock-in mutation leads to amyloid deposition in human wild-type amyloid precursor protein YAC transgenic mice Structural biology of presenilins and signal peptide peptidases The amyloid hypothesis of Alzheimer’s disease at 25 years Orthogonal gene knockout and activation with a catalytically active Cas9 nuclease Programmable base editing of A*T to G*C in genomic DNA without DNA cleavage Evolved Cas9 variants with broad PAM compatibility and high DNA specificity Improved base excision repair inhibition and bacteriophage Mu Gam protein yields C:G-to-T:A base editors with higher efficiency and product purity Transgenic mice with Alzheimer presenilin 1 mutations show accelerated neurodegeneration without amyloid plaque formation Hippocampal neuron loss exceeds amyloid plaque load in a transgenic mouse model of Alzheimer’s disease Murine Abeta over-production produces diffuse and compact Alzheimer-type amyloid deposits Single App knock-in mouse models of Alzheimer’s disease Genetic screening of Alzheimer’s disease genes in Iberian and African samples yields novel mutations in presenilins and APP Mechanism of the cleavage specificity of Alzheimer’s disease gamma-secretase identified by phenylalanine-scanning mutagenesis of the transmembrane domain of the amyloid precursor protein motif and transmembrane domain 9 of presenilin 1 are involved in the formation of the catalytic pore of the gamma-secretase C terminus of presenilin is required for overproduction of amyloidogenic Abeta42 through stabilization and endoproteolysis of presenilin Download references We thank Yukiko Nagai for secretarial assistance and Erika Sasaki and Wakako Kumita (Central Institute for Experimental Animals) for valuable discussions and advice This work was supported by AMED under Grant Number JP18dm0207001 (Brain Mapping by Integrated Neurotechnologies for Disease Studies (Brain/MINDS)) (T.C.S.) JSPS Grant-in-Aid for Scientific Research (C) Grant Number 18K07402 (H.S.) and the Special Postdoctoral ResearchersProgram from RIKEN (S.H.) including injections of BE and Target-AID into zygotes and embryo transfer were performed at the RIKEN CBS-Research Resource Division (RRD) We are grateful to the RIKEN Center for Brain Science RRD for technical help with DNA sequencing analyses These authors contributed equally: Hiroki Sasaguri Department of Neurology and Neurological Science Department of Molecular and Cellular Biology analyzed the data and prepared the figures All authors provided feedback and agreed on the final manuscript a RIKEN venture based at the Center for Brain Science a RIKEN venture based at the Brain Science Institute The remaining authors declare no competing interests Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations Download citation DOI: https://doi.org/10.1038/s41467-018-05262-w Anyone you share the following link with will be able to read this content: a shareable link is not currently available for this article Functional & Integrative Genomics (2024) Sign up for the Nature Briefing newsletter — what matters in science Your browser does not support JavaScript, or it is disabled．Please check the site policy for more information National Report a woman accused of being in a domineering relationship denied any role in the starvation death of her friend’s 5-year-old son or bilking his mother out of her money of controlling the life of the boy’s mother and not providing young Shojiro Ikari with enough food to live They were both separately indicted on charges of abandoning their guardian responsibilities resulting in death and Ikari was convicted in her own trial in June was found dead from starvation in April 2020 in Sasaguri But Akahori said during the lay judge hearing at the Fukuoka District Court on Aug 29 that she “did not give any instructions” that the boy’s diet should be restricted The defense maintained Akahori’s innocence and said she was asked by Ikari to cooperate in restricting the boy’s diet The two had met because their children attended the same kindergarten Ikari was “the first friend (Akahori) had in a long time,” the defense said so Akahori “kept company with (Ikari) because she did not want Ikari to dislike her.” According to the indictment and other sources Akahori and Ikari conspired to start limiting the amount of food Shojiro could eat from around August 2019 Akahori was also charged with fraud and theft for scamming Ikari out of about 2 million yen ($14,430) including her allowance for dependent children Akahori denied those charges as well at the hearing and said she was asked by Ikari to withdraw the cash The prosecution said in its opening statement that Akahori had repeatedly lied to Ikari telling her that the two were being sued by another mother Akahori told Ikari she needed the money to settle the dispute and scammed Ikari out of almost all the cash she had Akahori gradually took control over all aspects of Ikari’s life But the defense countered that it was a fictitious story The defense said Ikari was the one who asked Akahori to “cooperate in order to follow the rules,” which included restricting Shojiro’s diet The prosecution showed testimony from a kindergarten staff member his daily bento lunch became modest and the boy started becoming thin Shojiro often asked for seconds of the kindergarten-provided lunch and ate many of the potatoes provided for children during a kindergarten event The kindergarten thought something was wrong and sent a letter to Ikari Ikari is currently appealing her five-year prison sentence She is expected to appear at Akahori’s trial as a witness (This article was written by Naoki Nakayama and Daichi Itakura.) accused of starving 5-year-old boy to death Mother handed 5-year prison term for starving son 2-year-old girl left home alone for 11 hours dies from heatstroke Kanagawa boy returned to mother became 4th sibling to die nurse indicted over death of boy trapped in bus Rampant abuse reported at nursery where boy died in bus Information on the latest cherry blossom conditions Please right click to use your browser’s translation function.) A series based on diplomatic documents declassified by Japan’s Foreign Ministry Here is a collection of first-hand accounts by “hibakusha” atomic bomb survivors chefs and others involved in the field of food introduce their special recipes intertwined with their paths in life A series about Japanese-Americans and their memories of World War II In-house News and Messages No reproduction or republication without written permission Please view the main text area of the page by skipping the main menu. The page may not be displayed properly if the JavaScript is deactivated on your browser Some popular tourist sites in Japan are refusing entry to foreign tourists Caretakers at the sites have said this is because of the “bad manners” and “abhorrent actions” of visitors from abroad, the Asahi Shimbun reported Asahi Shimbun also reported that Nanzoin temple in Sasaguri has posted signs in more than 10 languages at its precincts and the nearby station describing Nanzoin as an important place of prayer and telling non-Japanese group travelers that they are not welcome Yatsushiro-gu shrine in Yatsushiro, Kumamoto Prefecture temporarily stopped accepting worshippers after the number of cruise ships arriving at a nearby port increased six-fold With further reports claiming the no-foreigners policy is spreading throughout Japan tourist sites risk the wrath of the country’s anti-discrimination laws and the United Nations’ International Convention on the Elimination of All Forms of Racial Discrimination was ordered to pay compensation for refusing to allow a naturalised Japanese citizen who was born in the United States to take a bath because of his appearance But according to a Justice Ministry official non-Japanese people who are denied entry to a place would have to report to the ministry to determine if the policy constitutes a human rights violation “It would be difficult for visiting tourists to file complaints,” the official said FUKUOKA--Two women in a "master-subordinate relationship" were arrested on March 2 here in connection with the starving death of one of the women's 5-year-old son abandoned their duties as guardians and left Ikari’s son Shojiro to die in April 2020 The women are accused of conspiring with each other to make Ikari’s third-born son malnourished by cutting down the frequency of his meals and the amount of food he received Police said the practice started around August 2019 and continued until the boy died at an apartment where Ikari lived in Sasaguri in the prefecture on April 18 found her son unconscious and asked a third party to call for an ambulance Shojiro looked extremely thin and weighed around 10 kilograms about half the average weight for a 5-year-old boy the boy’s thymus became atrophic when he died a sign that he was under severe stress likely caused by abuse Ikari told investigators that Akahori “sometimes hit Shojiro when he pushed back against her and did not obey what she told him to do.” police are also investigating the allegation that Akahori had assaulted the boy both physically and mentally for a long time Ikari and Akahori became acquainted when their children attended the same kindergarten in April 2016 Police said the unusual dependence relationship between the two women began with a manipulative lie in which Akahori told Ikari that her husband was having an affair “You have to maintain a simple lifestyle and practice frugal eating in order to win a lawsuit against your ex-husband,” police said Ikari at the time lived with her three sons started meddling in Ikari’s family affairs running the family’s household budgets and telling her how to raise her children Akahori gave precise instructions to Ikari on the amount of food to feed them “Ikari was under mind control,” an investigative source said Akahori regularly demanded money from Ikari for an “infidelity investigation on your former husband,” which she made up Ikari not only handed the livelihood protection subsidy and child care allowance she received to Akahori but also came up with extra cash by selling her car and borrowing from friends to give to the woman Ikari received a monthly income on welfare of around 200,000 yen Police believe that the total amount Ikari gave Akahori exceeded 10 million yen ($93,500) Akahori spent the rest of the money on clothes and other things police have concluded that Akahori was the de facto manager of the family budget and therefore had a responsibility to protect Shojiro when he became frail Akahori has denied the charge and told investigators “I had no involvement in the dietary management of the family.” Ikari has admitted to the charge and told investigators I couldn’t protect my (son) as his mother,” according to police Police did not find any external injuries on Ikari's three children but reported the case to a child consultation center a child consultation center took Ikari's two other children into protective custody (This article was written by Daichi Itakura and Tsubasa Yokoyama.) Record 193,780 child abuse cases responded to in Japan in FY2019 73 children die of abuse in Japan in FY2018 No stone left unturned in quest to probe all child deaths Abusive father’s ex-colleague sets out to help other abused children dies in Tokyo as mother visits boyfriend in Kyushu Ministry plans database of child-abusing baby-sitters Kyushu Grand Sumo Tournament winner Abi on Monday described his first career top-division championship as a long time coming To win a championship "was a dream when I was a rookie wrestler," the 28-year-old former sekiwake told an online press conference a day after winning the tournament at Fukuoka Kokusai Center in a rare three-way playoff "I began to realize how sweet it is to win a tournament when I returned to my room and took pictures with stablemates." a ninth-ranked maegashira for the just-ended tournament forced the playoff by defeating overnight leader and No 1 maegashira Takayasu to improve his record to 12-3 before beating Takayasu again and ozeki Takakeisho in the first three-way playoff since the 1994 Spring Grand Sumo Tournament could not travel to Fukuoka to see Abi's moment of victory due to health problems such as arrhythmias "I got this result after I followed what my boss told me: concentrate on one bout at a time," Abi said This year's makuuchi-division sumo saw different champions in all six tournaments: Mitakeumi Sumo: Abi wins 3-way playoff to claim maiden championship To have the latest news and stories delivered to your inbox Simply enter your email address below and an email will be sent through which to complete your subscription Please check your inbox for a confirmation email Thank you for reaching out to us.We will get back to you as soon as possible FUKUOKA--A mother who allowed her 5-year-old son to starve to death while in a relationship with a domineering older woman was found guilty of abandoning her parental responsibilities and sentenced to five years in prison The Fukuoka District Court’s June 17 decision for Rie Ikari took into account the fact that she herself was a victim because her actions were under the control of Emiko Akahori The latter has been indicted separately on a charge of abandoning her guardian responsibilities resulting in death was found dead in their apartment in Sasaguri “The despair that must have been felt by the victim is unfathomable He did not receive sufficient protection from the one person he should have been able to depend upon The court determined that Ikari was unable to think coherently because she herself was deprived of food and sleep while under Akahori’s control Although the court conceded that Ikari felt she had no choice but to follow Akahori’s rules it also said the defendant deserved censure for failing to fulfill her responsibility by turning to relatives for help in keeping her son safe you will stand by the growth of (her two other sons) Please live strongly until that day arrives.” The case was heard by citizen judges and four agreed to a news conference after the verdict was read admitted it was difficult to comprehend the control exerted by Akahori as it was so far removed from what would be considered normal in daily life an associate professor of clinical psychology at Tokyo Future University said a person in Ikari’s situation would be less than coherent in a relationship based on dominance coupled with a lack of sufficient food and sleep Iume speculated that Akahori might have been able to reinforce her hold on Ikari by instilling fear in the woman and causing her to distrust others around her (This article was written by Naoki Nakayama Husband of woman slain in Fukuoka tries to help others FUKUOKA--The Fukuoka District Court on Sept 21 handed down a 15-year prison sentence sought by prosecutors to Emiko Akahori over the starving death of her friend’s son Akahori was found guilty of being an accomplice in the crime of abandoning guardian responsibilities resulting in the death of Shojiro Ikari The court found that Akahori domineered over every aspect of Ikari’s life and told her to feed her boy less he was fed only a tiny amount of food,” the court said “He could not enjoy the protection of his mother whom he should have been able to depend on The court said Akahori “knew that Shojiro was malnourished but did not end her psychological control (over his mother) and continued to not offer protection for him.” That made Akahori an accomplice with Ikari who is currently appealing her five-year prison sentence after being found guilty of abandoning her parental responsibilities in connection with Shojiro's death Akahori also swindled about 2 million yen ($13,700) from Ikari under the pretense of fees for investigating her husband’s affairs and other things Akahori denied the charges and proclaimed her innocence Friend of mother denies role in starving young boy to death Man sentenced to 7 years in prison over sex abuse of stepdaughter Mother fed newborn her blood to get better medical help The issue of women from other countries being forced into sexual servitude as "comfort women" under the Japanese military during wartime has been well documented But the voices of Japanese women who say they were sexually exploited by their countrymen have rarely been heard "For the first time we focus on Japanese women despite their painful experiences during World War II," said Mina Watanabe secretary general of the Women's Active Museum on War and Peace in Tokyo which has organized several exhibitions about comfort women since its launch in 2005 by displaying official historical documents and correspondence from Japanese soldiers as well as testimonies of the former comfort women that Japanese comfort women were procured by various means while others were deceived into service as prostitutes in 1937 after working at a licensed quarter in Japan After going through medical checkups for sexually transmitted diseases Sasaguri accompanied Japanese troops for six years from China "We will lose our prestige if it is widely known that the Imperial Army accompanied such women Kanayo Kurose was born into a poor family of tenant farmers who became indebted after their rice crop business was damaged by typhoons and an economic slump where she helped recruit Japanese and Korean women "We should say it was illegal to detain them in sexual servitude even if they were indebted," Watanabe said One woman gives testimony of traveling to Taiwan in 1941 at the age of 15 as she was asked to help a hotel operation there she was transferred to Manila on the order of the military where she was exploited as a comfort woman "I didn't expect to be used as a comfort woman," she says in a panel display She was required to regularly undergo medical checkups Others were tricked into believing that they could qualify as nurses if they volunteered unaware that they would be forced to have sex at military brothels a civilian worker for the military notes in a panel display it is impossible to generalize about Japanese comfort women and to paint their whole picture," Watanabe said "We present only fragments of their suffering." Comfort stations were also located in many parts of Japan and similar operations continued even after the end of the war An organization known as the Recreation and Amusement Association set up a comfort station to provide organized prostitution in Tokyo less than two weeks after the war to serve the occupying U.S "It reflected an assumption that soldiers require comfort stations soldiers would do the same thing the Japanese soldiers had done -- rape in occupied areas," Watanabe said resulting in many of the Japanese women who found themselves suddenly unemployed going on to become street prostitutes The exhibition suggests most victimized Japanese women have had no other choice but to remain silent because of the notion that they are responsible for being targets of sexual exploitation "They have been led to feel bad about what they went through and have been hesitant to come out to struggle for their rights," Watanabe said One person who is still alive decided against displaying a testimonial after once approving it because she feared being exposed as a former comfort woman Japan and South Korea agreed in 2015 to "finally and irreversibly" resolve the longstanding comfort women issue Japan has demanded the removal of statues symbolizing Korean comfort women installed in front of Japanese diplomatic offices Watanabe said the issue is not only a bilateral one between Japan and South Korea "It is necessary for Japan to consider legislation so it provides reparations to victimized women regardless of their nationalities." Taiwan remembers vanishing "comfort women" Several Japanese attractions are turning away foreign tourists or those visiting in large groups This content can also be viewed on the site it originates from some don't flush toilets after use and some even leave chewing gum on the soles of the Buddha's feet Signs around town and in the trains of Sasaguri now explain that Nanzo-in is a place of worship and that non-Japanese tourists are not welcome And even Japanese tourists who misbehave will be turned away This content can also be viewed on the site it originates from stopped accepting tourists temporarily on days cruise ships arrived at the port after the government deployed staff to give instructions to tourists on how to respect and obey Japanese customs Use of this website constitutes acceptance of our Terms of Service (updated April 1 2021) and Policy and Cookie Statement (updated April 1 The material on this site may not be reproduced except with the prior written permission of Condé Nast Japanese version Metrics details Histidine decarboxylase is one of the enzymes of the amine precursor uptake and decarboxylation system and is known to be distributed in mast cells and enterochromaffin-like cells With the hypothesis that histidine decarboxylase expression is a marker for neuroendocrine differentiation we studied the immunoreactivity of histidine decarboxylase in neuroendocrine cells and tumors of the thyroid gland Formalin-fixed paraffin sections were subjected to immunohistochemistry using anti-histidine decarboxylase antibody and the sensitivity and specificity were compared with those of conventional neuroendocrine markers (CD56 Enterochromaffin or enterochromaffin-like cells and thyroid C-cells were positive for histidine decarboxylase medullary carcinoma) showed a high percentage of positive staining we used the antibody to distinguish small cell lung carcinoma from non-neuroendocrine lung carcinoma and also to detect neuroendocrine differentiation in large-cell neuroendocrine carcinoma and gastrointestinal small-cell carcinoma The anti-histidine decarboxylase antibody stained most small cell lung carcinoma (18 of 23 and was rarely reactive with non-neuroendocrine lung tumors (2 of 44; specificity These values were close to those obtained from CD56 staining (sensitivity/specificity Histidine decarboxylase was also positive for 6 of 12 large cell neuroendocrine carcinomas and 4 of 7 gastrointestinal small cell carcinomas we demonstrated that histidine decarboxylase is useful to distinguish between small cell lung carcinoma and non-neuroendocrine carcinoma and to demonstrate neuroendocrine differentiation no systematic immunohistochemical study of HDC expression in neuroendocrine or endocrine tumors has been reported We planned this study to screen the expression of HDC in lung and gastrointestinal neuroendocrine tumors we compared the sensitivity and specificity of HDC staining with those of widely used neuroendocrine markers The diagnosis of small cell lung carcinoma and other carcinoma is based primarily on light-microscopic observation Twenty-three cases of carcinoid tumors and adenocarcinoma/squamous cell carcinomas with small cell carcinoma or neuroendocrine carcinoma components of the gastrointestinal tract were also examined 15 cases of adrenal pheochromocytomas and 4 cases of medullary carcinomas of the thyroid gland were also selected The diagnosis of all these cases was confirmed by at least three board-certified surgical pathologists The second peroxidase-labeled antibody was applied and incubated with a solution of 20 mg of 3,3′-diaminobenzidene tetrahydrochloride and 20 μL of H2O2 in 100 mL of Tris-HCl (50 mm After counterstaining with Meyer’s hematoxylin the sections were observed by light microscopy staining was considered to be positive if ≥5% of tumor cells were reactive and was classified into two groups: (1) focally positive staining in <10% of tumor cells and (2) strongly positive staining in ≥10% of tumor cells All pheochromocytomas (15 cases) and medullary carcinomas of the thyroid gland (4 cases) tested showed diffuse and strong cytoplasmic immunoreactivity with anti-HDC antibody (Figures 1A,B), as did normal adrenal chromaffin cells and thyroid gland C-cells (not shown). Immunohistochemical demonstration of HDC in pheochromocytoma (A) Diffuse cytoplasmic staining of HDC is evident No HDC staining was observed in squamous cell carcinomas (0 of 21) and adenosquamous cell carcinomas (0 of 3) 7 of 72 non-small and non-neuroendocrine tumors (9.7%) HDC staining in gastrointestinal neuroendocrine cells and tumors Scattered HDC-positive cells are located in the gastric (A) and colonic (B) mucosa (C) The cells of duodenal carcinoid tumor are strongly reactive with anti-HDC antibody (D) Rectal cancer admixed with adenocarcinoma and small cell carcinoma Note that HDC is positive only in the small cell carcinoma component Results of staining for NSE, SYN, CGA, and CD56 are shown in Table 1 and bronchial and gastrointestinal-carcinoid tumors With regard to other lung and gastrointestinal carcinomas an immunohistochemical panel was used to compare the sensitivity and specificity of HDC with those of other neuroendocrine markers Staining for CD56 showed the highest (87% positive) score and all the large cell neuroendocrine carcinomas were positive for at least one of the neuroendocrine markers large-cell carcinoma lacked positive staining for CD56 one case of adenocarcinoma that was focally positive for HDC was also focally positive for CD56 Another case each of adenosquamous cell carcinoma and squamous cell carcinoma were focally reactive for SYN The percentage of positive staining for NSE was 41% in adenocarcinomas one adenocarcinoma was focally positive for both NSE and HDC positive staining for the panel of antibodies was similar to that of large cell neuroendocrine carcinomas in the lung and CGA and SYN were reactive in 43% (3 of 7) and 57% (4 of 7) cases Among the small cell lung carcinoma, one case was negative for all markers including HDC. On the other hand, three cases were positive for all markers. In Table 2 we summarized the frequency of the reactivity of five neuroendocrine markers in the small cell lung carcinoma Seventy-eight percent of cases (18/23) of the small cell carcinoma were positive for three or more neuroendocrine markers and 94% of HDC-positive cases (17/18) were also reactive to the other two or more markers we evaluated the possibility that HDC expression is a novel neuroendocrine differentiation marker we investigated HDC expression in non-tumor neuroendocrine cells and gastrointestinal and bronchial neuroendocrine cells were strongly positive for HDC we expanded the application to neuroendocrine tumors which also showed intense and diffuse staining This immunohistochemical demonstration of HDC in normal neuroendocrine cells and neuroendocrine tumors as a positive control indicated that HDC is an excellent marker for neuroendocrine differentiation but no neuroendocrine morphology; (3) large cell carcinoma with neuroendocrine morphology without neuroendocrine differentiation; and (4) classic large cell carcinoma with no neuroendocrine morphology or differentiation by special studies including immunohistochemistry or electron microscope (EM) and we had no cases with the appearance of Category 2 or 3 if we use the immunohistochemical results to identify neuroendocrine differentiation If we consider that HDC is a useful marker to of neuroendocrine differentiation the five large cell carcinoma cases may be categorized as large cell carcinoma with neuroendocrine differentiation showing no neuroendocrine morphology four of the five HDC-positive large cell carcinomas were also reactive for NSE These two markers showed a relatively higher sensitivity compared with CGA and SYN The difference in subcellular localization of marker proteins may influence their detectability we demonstrated that HDC is a useful diagnostic tool for distinction between small cell lung carcinoma and non-neuroendocrine carcinoma in combination with other conventional markers Detection of chromogranin in neuroendocrine cells with a monoclonal antibody Immunohistology of neuroendocrine and neuroectodermal tumors Chromogranin as a marker of neuroendocrine cells in cytologic material—an immunocytochemical study and chromogranin as markers for neuroendocrine lung tumors Immunoelectron microscopy for chromogranin A in small cell neuroendocrine carcinoma of lung Immunohistochemical localization of gamma-enolase in normal human tissue other than nervous and neuroendocrine tissues Utility of 123C3 monoclonal antibody against CD56 (NCAM) for the diagnosis of small cell carcinomas on paraffin sections Microtubule-associated protein-2: a new sensitive and specific marker for pulmonary carcinoid tumor and small cell carcinoma Intracellular localization of the 74- and 53-kDa forms of l-histidine decarboxylase in a rat basophil/mast cell line Electron microscopic identification of histidine decarboxylase-containing endocrine cells of the rat gastric mucosa Increase in histidine decarboxylase activity in skin of genetically mast-cell-deficient W/W mice after application of phorbol 12-myristate 13-acetate: evidence for the presence of histamine-producing cells without basophilic granules Proc Natl Acad Sci U S A 1982; 79: 6837–6841 Effects of histamine and interleukin-4 synthesized in arterial intima on phagocytosis by monocytes/macrophages in relation to atherosclerosis Histamine: an early messenger in inflammatory and immune reactions Molecular biology and role of histamine in physiological and pathological reactions Analysis of vasoconstrictor responses to histamine in the hind limb vascular bed of the rabbit Gastric carcinoid with histamine production histamine transporter and expression of somatostatin receptors Cimetidine inhibits in vivo growth of human colon cancer and reverses histamine stimulated in vitro and in vivo growth Effect of cimetidine on intratumoral cytokine expression in an experimental tumor Biochem Biophys Res Commun 2001; 281: 1113–1116 Suppression of melanoma cell proliferation by histidine decarboxylase specific antisense oligonucleotides The amine precursor uptake and decarboxylation cell concept and its implications in pathology The amine precursor uptake and decarboxylation cell system and its neoplasm: observation on the significance and limitations of the concept and l-dopa decarboxylase as markers for neuroendocrine differentiation in lung cancer cell lines Expression of neuroendocrine cell markers l-dopa decarboxylase and dense core granules in human tumors of endocrine and neuroendocrine origin Expression of histidine decarboxylase and synthesis of histamine by human small cell lung carcinoma World Health Organization Histological typing of lung and pleural tumors Neuroendocrine tumors of the lung with proposed criteria for large-cell neuroendocrine carcinoma Immunohistochemical localization of aromatic l-amino acid decarboxylase in human and mouse bronchopulmonary and gastrointestinal endocrine cells Immunohistochemical study of small cell carcinoma with special reference to the neuroendocrine markers aromatic l-amino acid decarboxylase and gastrin-releasing peptide Immunohistologic analysis of gastrointestinal and pulmonary carcinoid tumors Immunostaining in the diagnosis of pulmonary neuroendocrine carcinomas: an immunohistochemical study with ultrastructural correlations Immunoreactivity for epithelial and neuroendocrine antibodies are useful in the differential diagnosis of lung carcinomas Neuroendocrine differentiation in endocrine and neuroendocrine lung carcinomas The neural cell adhesion molecule (NCAM) as a regulator of cell-cell interactions Download references University of Occupational and Environmental Health Download citation DOI: https://doi.org/10.1097/01.MP.0000044485.14910.3A Journal of Cancer Research and Clinical Oncology (2017)