Cell adhesion molecule 1 (CADM1), a single-pass transmembrane protein, is involved in oncogenesis. We previously demonstrated the therapeutic efficacy of anti-CADM1 ectodomain monoclonal antibodies against mesothelioma; however, the underlying mechanism is unclear. In the present study, we explored the molecular behavior of anti-CADM1 antibodies in CADM1-expressing tumor cells. Sequencing analyses revealed that the anti-CADM1 chicken monoclonal antibodies 3E1 and 9D2 are IgY and IgM isotype antibodies, respectively. Co-administration of 3E1 and 9D2 altered the subcellular distribution of CADM1 from the detergent-soluble fraction to the detergent-resistant fraction in tumor cells. Using recombinant chicken-mouse chimeric antibodies that had been isotype-switched from IgG to IgM, we demonstrated that the combination of the variable region of 3E1 and the constant region of IgM was required for CADM1 relocation. Cytochemical studies showed that 3E1 colocalized with late endosomes/lysosomes after co-administration with 9D2, suggesting that the CADM1-antibody complex is internalized from the cell surface to intracellular compartments by lipid-raft mediated endocytosis. Finally, 3E1 was conjugated with the antimitotic agent monomethyl auristatin E (MMAE) via a cathepsin-cleavable linker. Co-administration of 3E1-monomethyl auristatin E and 9D2 suppressed the growth of multiple types of tumor cells, and this anti-tumor activity was confirmed in a syngeneic mouse model of melanoma. 3E1 and 9D2 are promising drug delivery vehicles for CADM1-expressing tumor cells.

Aims: Cell adhesion molecule 1 (CADM1) mediates interepithelial adhesion and is upregulated in crowded epithelial monolayers. This study aimed to examine CADM1 expression in the human endometrium of proliferative and secretory phases, and its transcriptional regulation in terms of estrogen stimuli and higher cellularity. Main methods: CADM1 immunohistochemistry was conducted on endometrial tissues from women in their 40s and adult mice subcutaneously injected with estradiol following ovariectomy. Dual-luciferase reporter assays were conducted using human endometrial HEC-50B and HEC-1B cells and reporter plasmids harboring the human CADM1 3.4-kb promoter and its deleted and mutated forms. Cells were transfected with estrogen receptor alpha cDNA and reporter plasmids, and treated with estradiol before luciferase activity measurement. Key findings: Immunohistochemistry revealed that CADM1 was clearly expressed on the lateral membranes of the simple columnar glandular cells in the proliferative phase, but not in the secretory phase, from both women and the mouse model. The glandular cell density increased two-fold in the proliferative phase. Reporter assays identified three Sp1-binding sites as estradiol-responsive elements in the proximal region (from -223 to -84) of the transcription start site (+1) in HEC-50B cells. When the cell culture was started at eight-fold higher cell density, the CADM1 3.4-kb promoter was transactivated at a two-fold higher level in HEC-50B cells. This cell density effect was not detected for the CADM1 2.3-kb or 1.6-kb promoter. Significance: Two (proximal and distal) promoter regions are suggested to function additively to transactivate CADM1 in endometrial glandular cells that crowd in the proliferative phase.

Aims: Stroke-prone spontaneously hypertensive rats (SHRSP) show significantly lower body weight than normotensive Wistar-Kyoto rats (WKY). Our hypotheses are as follows: weight loss of the skeletal muscle is related to hypertension-related diseases, and muscle hypotrophy is useful as a therapeutic target for hypertension and hypertension-related diseases. In this study, we aimed to investigate the pathophysiological characteristics of muscle hypotrophy in SHRSP to determine the therapeutic target molecule(s). Main methods: The difference in skeletal muscles in the lower leg between WKY and SHRSP was evaluated mainly through weight/tibial length, histological, gene expression, and protein expression analyses. Key findings: SHRSP had a significantly lower weight/tibial length in soleus and gastrocnemius, but not in plantaris and tibialis anterior, indicating that muscles consisting of a relatively high amount of slow muscle fiber were affected. This result was confirmed by the histological analysis of soleus, showing that type I fiber mainly decreased the fiber size. Microarray and protein expression analyses showed that the muscle-specific ubiquitin ligase, muscle RING finger 1 (MuRF1), but not atrogin-1, was highly expressed in soleus, but not in plantaris, in SHRSP. TNF-like weak inducer of apoptosis receptor (TWEAKR) was predicted as a MuRF1 up-regulator by Ingenuity Pathway Analysis and immunostained only in type II fiber in WKY but in both type I and II fibers in SHRSP. Significance: TWEAKR is a type II-specific receptor in the skeletal muscle. Ectopic TWEAKR expression in type I fiber of SHRSP is most likely involved in slow muscle-specific hypotrophy through MuRF1 overexpression.

Elevation of intraocular pressure is a major risk factor for glaucoma development, which causes the loss of retinal ganglion cells (RGCs). Lipocalin 2 (Lcn2) is upregulated in glaucomatous retinae; however, whether Lcn2 is directly involved in glaucoma is debated. In this study, retinal explant cultures were subjected to increased water pressure using a two-chamber culture device, and Lcn2 protein levels were examined by immunoblotting. In situ TdT-mediated dUTP nick and labeling (TUNEL) and glial fibrillary acidic protein (GFAP) immunohistochemical assays were performed to assess apoptosis and gliosis, respectively. The neurotoxicity of Lcn2 in the retinal explant culture was determined with exogenous administration of recombinant Lcn2. The Lcn2 protein levels, percentage of TUNEL-positive cells, and GFAP-positive area were significantly higher in retinae cultured under 50 cm H 2 O pressure loads compared to those cultured under 20 cm H 2 O. We found that Lcn2 exhibited neurotoxicity in retinae at dose of 1 μg/ml. The negative effects of increased hydrostatic pressure were attenuated by the iron chelator deferoxamine. This is the first report demonstrating the direct upregulation of Lcn2 by elevating hydrostatic pressure. Modulating Lcn2 and iron levels may be a promising therapeutic approach for retinal degeneration.

When epithelial cells in vivo are stimulated to proliferate, they crowd and often grow in height. These processes are likely to implicate dynamic interactions among lateral membranous proteins, such as cell adhesion molecule 1 (CADM1), an immunoglobulin superfamily member. Pulmonary epithelial cell lines that express CADM1, named NCI-H441 and RLE-6TN, were grown to become overconfluent in the polarized 2D culture system, and were examined for the expression of CADM1. Western analyses showed that the CADM1 expression levels increased gradually up to 3 times in a cell density-dependent manner. Confocal microscopic observations revealed dense immunostaining for CADM1 on the lateral membrane. In the overconfluent monolayers, CADM1 knockdown was achieved by two methods using CADM1-targeting siRNA and an anti-CADM1 neutralizing antibody. Antibody treatment experiments were also done on 6 other epithelial cell lines expressing CADM1. The CADM1 expression levels were reduced roughly by half, in association with cell height decrease by half in 3 lines. TUNEL assays revealed that the CADM1 knockdown increased the proportion of TUNEL-positive apoptotic cells approximately 10 folds. Increased expression of CADM1 appeared to contribute to cell survival in crowded epithelial monolayers.

As one of the contributions of this paper, this paper first explores the upper bound of context-based neural machine translation and attempt to utilize previously un-used context information. We found that, if we could appropriately select the most informative context sentence for a given input source sentence, we could boost translation accuracy as much as approximately 10 BLEU points. This paper next explores a criterion to select the most informative context sentences that give the highest BLEU score. Applying the proposed criterion, context sentences that yield the highest forced back-translation probability when back-translating into the source sentence are selected. Experimental results with Japanese and English parallel sentences from the OpenSubtitles2018 corpus demonstrate that, when the context length of five preceding and five subsequent sentences are examined, the proposed approach achieved significant improvements of 0.74 (Japanese to English) and 1.14 (English to Japanese) BLEU scores compared to the baseline 2-to-2 model, where the oracle translation achieved upper bounds improvements of 5.88 (Japanese to English) and 9.10 (English to Japanese) BLEU scores.

In this paper, we propose a multi-hop attention for the Transformer. It refines the attention for an output symbol by integrating that of each head, and consists of two hops. The first hop attention is the scaled dot-product attention which is the same attention mechanism used in the original Transformer. The second hop attention is a combination of multi-layer perceptron (MLP) attention and head gate, which efficiently increases the complexity of the model by adding dependencies between heads. We demonstrate that the translation accuracy of the proposed multi-hop attention outperforms the baseline Transformer significantly, +0.85 BLEU point for the IWSLT-2017 German-to-English task and +2.58 BLEU point for the WMT-2017 German-to-English task. We also find that the number of parameters required for a multi-hop attention is smaller than that for stacking another self-attention layer and the proposed model converges significantly faster than the original Transformer.

The Breast JournalVolume 26, Issue 2 p. 269-270 BREAST IMAGES Phyllodes tumor with extensive intraductal spread radiologically mimicking ductal carcinoma in situ Tomoyuki Otani MD, Corresponding Author tomoyu@gmail.com orcid.org/0000-0002-4351-324X Department of Pathology, Kindai University Faculty of Medicine, Osaka-Sayama, Japan Correspondence Tomoyuki Otani, Department of Pathology, Kindai University Faculty of Medicine, 377-2 Ohno-Higashi, Osaka-Sayama 589-8511, Osaka, Japan. Email: tomoyu@gmail.comSearch for more papers by this authorMika Hamada MD, Division of Breast and Endocrine Surgery, Department of Surgery, Kindai University Faculty of Medicine, Osaka-Sayama, JapanSearch for more papers by this authorYukihiko Hashimoto MD, Division of Breast and Endocrine Surgery, Department of Surgery, Kindai University Faculty of Medicine, Osaka-Sayama, JapanSearch for more papers by this authorChihiro Higashi MD, Division of Breast and Endocrine Surgery, Department of Surgery, Kindai University Faculty of Medicine, Osaka-Sayama, JapanSearch for more papers by this authorHirofumi Kanaizumi MD, Division of Breast and Endocrine Surgery, Department of Surgery, Kindai University Faculty of Medicine, Osaka-Sayama, JapanSearch for more papers by this authorYumiko Tanaka MD, orcid.org/0000-0001-5042-5208 Division of Breast and Endocrine Surgery, Department of Surgery, Kindai University Faculty of Medicine, Osaka-Sayama, JapanSearch for more papers by this authorWataru Shinzaki MD, Division of Breast and Endocrine Surgery, Department of Surgery, Kindai University Faculty of Medicine, Osaka-Sayama, JapanSearch for more papers by this authorRyuichiro Kimura PhD, Department of Pathology, Kindai University Faculty of Medicine, Osaka-Sayama, JapanSearch for more papers by this authorYoshifumi Komoike MD, PhD, Division of Breast and Endocrine Surgery, Department of Surgery, Kindai University Faculty of Medicine, Osaka-Sayama, JapanSearch for more papers by this authorAkihiko Ito MD, PhD, Department of Pathology, Kindai University Faculty of Medicine, Osaka-Sayama, JapanSearch for more papers by this author Tomoyuki Otani MD, Corresponding Author tomoyu@gmail.com orcid.org/0000-0002-4351-324X Department of Pathology, Kindai University Faculty of Medicine, Osaka-Sayama, Japan Correspondence Tomoyuki Otani, Department of Pathology, Kindai University Faculty of Medicine, 377-2 Ohno-Higashi, Osaka-Sayama 589-8511, Osaka, Japan. Email: tomoyu@gmail.comSearch for more papers by this authorMika Hamada MD, Division of Breast and Endocrine Surgery, Department of Surgery, Kindai University Faculty of Medicine, Osaka-Sayama, JapanSearch for more papers by this authorYukihiko Hashimoto MD, Division of Breast and Endocrine Surgery, Department of Surgery, Kindai University Faculty of Medicine, Osaka-Sayama, JapanSearch for more papers by this authorChihiro Higashi MD, Division of Breast and Endocrine Surgery, Department of Surgery, Kindai University Faculty of Medicine, Osaka-Sayama, JapanSearch for more papers by this authorHirofumi Kanaizumi MD, Division of Breast and Endocrine Surgery, Department of Surgery, Kindai University Faculty of Medicine, Osaka-Sayama, JapanSearch for more papers by this authorYumiko Tanaka MD, orcid.org/0000-0001-5042-5208 Division of Breast and Endocrine Surgery, Department of Surgery, Kindai University Faculty of Medicine, Osaka-Sayama, JapanSearch for more papers by this authorWataru Shinzaki MD, Division of Breast and Endocrine Surgery, Department of Surgery, Kindai University Faculty of Medicine, Osaka-Sayama, JapanSearch for more papers by this authorRyuichiro Kimura PhD, Department of Pathology, Kindai University Faculty of Medicine, Osaka-Sayama, JapanSearch for more papers by this authorYoshifumi Komoike MD, PhD, Division of Breast and Endocrine Surgery, Department of Surgery, Kindai University Faculty of Medicine, Osaka-Sayama, JapanSearch for more papers by this authorAkihiko Ito MD, PhD, Department of Pathology, Kindai University Faculty of Medicine, Osaka-Sayama, JapanSearch for more papers by this author First published: 19 September 2019 https://doi.org/10.1111/tbj.13545Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinked InRedditWechat No abstract is available for this article. Volume26, Issue2February 2020Pages 269-270 RelatedInformation

Cell adhesion molecule 1 (CADM1) is an immunoglobulin superfamily member strongly expressed on renal tubular epithelia in the urinary tract. Enzymatic cleavage of its ectodomain increases in chronic kidney disease (CKD), and is assumed to contribute to tubulointerstitial lesion formation. Because the cleaved ectodomain fragments are likely to be released into the urine, a sandwich enzyme-linked immunosorbent assay (ELISA) system for urinary CADM1 was developed using two anti-ectodomain antibodies. Urinary CADM1 concentrations in patients with CKD based on various forms of glomerulonephritis and nephropathy (n = 127) were measured. A total of 44 patients (35%) had elevated CADM1 concentrations over the normal upper limit (362 pg/mL), with a mean of 1,727 pg/mL. Renal biopsy specimens of all patients were pathologically scored for tubulointerstitial lesions using epithelial degeneration, interstitial inflammation, and fibrosis. There were no correlations between urinary CADM1 concentrations and pathological scores or any widely used renal markers, including glomerular filtration rate (GFR), but there was a weak inverse correlation between pathological scores and GFR (R2 = 0.292). Notably, this correlation gradually increased in patients with increasing CADM1 concentrations, and reached a maximum R 2 (0.899) at a cutoff of 1,569 pg/mL. The results of this study suggest that urinary CADM1 is a useful marker indicating tubulointerstitial damage from elevated GFR levels in CKD.

Among recent progresses of neural machine translation models, the invention of the Transformer model is one of the most important progresses. It is well-known that the key technologies of the Transformer include multi-head attention mechanism. This paper introduces the multi-head attention mechanism into the traditional RNN-based neural machine translation model. Moreover, inspired by the existing multi-hop architectures such as end-to-end memory networks and convolutional sequence to sequence learning model, this paper proposes an RNN based NMT model with a multi-hop attention mechanism. The proposed multi-hop attention model has two heads, where for each head, a context vector is calculated based on the states of the encoder and the decoder. Then, in the second turn of the context vector calculation, those context vectors are updated depending not only on one’s own context vector but also on the context vector of the other head. Experimental re-sults show that the proposed model signifi-cantly outperforms the baseline in BLEU score in Japanese-to-English/English-to-Japanese machine translation tasks with and without extended context.

Polypeptide N‑acetylgalactosaminyltransferase 6 (GALNT6), which is involved in the initiation of O‑glycosylation, has been reported to play crucial roles in mammary carcinogenesis through binding to several substrates; however, its biological roles in mediating growth‑promoting effects remain unknown. The present study demonstrated a crucial pathophysiological role of GALNT6 through its O‑glycosylation of lectin galactoside‑binding soluble 3 binding protein (LGALS3BP), a secreted growth‑promoting glycoprotein, in breast cancer growth. The Cancer Genome Atlas data analysis revealed that high expression levels of GALNT6 were significantly associated with poor prognosis of breast cancer. GALNT6 O‑glycosylated LGALS3BP in breast cancer cells, whereas knockdown of GALNT6 by siRNA led to the inhibition of both the O‑glycosylation and secretion of LGALS3BP, resulting in the suppression of breast cancer cell growth. Notably, LGALS3BP is potentially O‑glycosylated at three sites (T556, T571 and S582) by GALNT6, thereby promoting autocrine cell growth, whereas the expression of LGALS3BP with three Ala substitutions (T556A, T571A and S582A) in cells drastically reduced GALNT6‑dependent LGALS3BP O‑glycosylation and secretion, resulting in suppression of autocrine growth‑promoting effect. The findings of the present study suggest that the GALNT6‑LGALS3BP axis is crucial for breast cancer cell proliferation and may be a therapeutic target and biomarker for mammary tumors.

あらまし ニューラル機械翻訳 (NMT) の弱点の一つとして,扱える語彙に限りがある点が知られている. NMTにおいては,語彙辞書に含まれていない単語は未知語トークンとして出力されるため,これが誤訳となる. 従来法では,出力文に含まれた未知語トークンが対応する原言語の単語を推定しその訳語に置き換えることに よって,NMT において出力可能となる語彙の規模を拡大した.しかし,この方式は,単語単位での語彙規模の 拡大にとどまる点が弱点であった.本論文においては,ニューラル翻訳において,大規模フレーズ語彙に対応す る方式を提案する.具体的には,訓練用対訳文においてフレーズ間の二言語対応の情報を収集し,二言語間で対 応済みのフレーズ対訳対を同一のトークンに置き換えた後,NMT モデルの訓練を行う.翻訳時には,NMT モ デルの語彙集合中の語彙部分に対しては,NMT モデルによる訳文生成がなされ,一方,その他のフレーズまた は単語語彙部分に対しては,SMT モデルによる翻訳がなされる.日中,中日,日英,英日の各方向の翻訳にお いて評価を行い,提案手法の有効性を検証した. キーワード NMT,SMT,大語彙,フレーズ翻訳

AIMS:To determine cellular distribution of cell adhesion molecule 1 (CADM1), an immunoglobulin superfamily member, in the human oxyntic gastric mucosa, and to explore possible involvement in the development and peritoneal dissemination of signet ring cell (SRC) gastric carcinoma, which often develops in the oxyntic mucosa.MAIN METHODS:Immunohistochemistry and double immunofluorescence were conducted on surgical specimens of normal and SRC-bearing stomachs and peritoneal metastatic foci of SRCs. KATO-III (lacking CADM1) and HSC-43 (expressing CADM1) SRC cell lines were cocultured on a Met-5A mesothelial or TIG-1 fibroblastic cell monolayer.KEY FINDINGS:In the oxyntic gland, some neck and nearly all base glandular cells were CADM1-positive, and mucin 5AC-positive cells were CADM1-negative, while some mucin 6-positive neck cells were CADM1-positive. Foveolar-epithelial, parietal, and endocrine cells were CADM1-negative. CADM1 was negative in all SRC carcinomas that were confined within the submucosa (n = 11) and all but one of those invading deeper (n = 15). In contrast, peritoneal metastatic foci of SRCs were CADM1-positive in five out of eleven cases (P < 0.01). In the cocultures, exogenous CADM1 made KATO-III cells adhere more and grow faster on a Met-5A monolayer, not on TIG-1 monolayers. HSC-43 cells adhered more and grew faster on Met-5A than on TIG-1 monolayers, which were partly counteracted by a function-neutralizing anti-CADM1 antibody.SIGNIFICANCE:Nearly all chief cells and a part of mucous neck cells express CADM1. SRC gastric carcinoma appears to emerge as a CADM1-negative tumor, but CADM1 may help SRCs develop peritoneal dissemination through promoting their adhesion and growth in the serosal tissue.

Neural machine translation (NMT), a new approach to machine translation, has achieved promising results comparable to those of traditional approaches such as statistical machine translation (SMT). Despite its recent success, NMT cannot handle a larger vocabulary because the training complexity and decoding complexity proportionally increase with the number of target words. This problem becomes even more serious when translating patent documents, which contain many technical terms that are observed infrequently. Long et al. (2016) proposed a method that enables NMT to translate patent sentences comprising a large vocabulary of technical terms. The proposed NMT system is trained on bilingual data wherein technical terms are replaced with technical term tokens; this allows it to translate most of the source sentences except technical terms. The selected phrases are then replaced with tokens during training and post-translated by the phrase translation table of SMT. Based on the discussion as well as experimental evaluation results reported in Long et al. (2016), this paper further studies the effect of the proposed NMT model with phrase translation by the SMT model with respect to reducing untranslated content. The issue of untranslated content is among those most important problems of NMT. This paper employs the back translation probability which is proposed by Goto and Tanaka (2017) to apply to the task of detecting untranslated content in NMT. Then, we show the evaluation results of both predicting untranslated contents and of manually counting the numbers of words in the input Japanese sentences untranslated into English in the task of Japanese to English NMT, where the proposed NMT model with phrase translation by the SMT model outperforms the baseline NMT model.