Aberrant expression of claudin-6 contributes to malignant potentials and drug resistance of cervical adenocarcinoma.

Recent studies have revealed that aberrant expression of tight junction (TJ) proteins is a hallmark of various solid tumors and it is recognized as a useful therapeutic target. Claudin-6 (CLDN6), a member of the family of TJ transmembrane proteins, is an ideal therapeutic target because it is not expressed in human adult normal tissues. In this study, we found that CLDN6 is highly expressed in uterine cervical adenocarcinoma (ADC) and that high CLDN6 expression was correlated with lymph node metastasis and lymphovascular infiltration and was an independent prognostic factor. Shotgun proteome analysis revealed that cell-cell adhesion-related proteins and drug metabolism-associated proteins (aldo-keto reductase [AKR] family proteins) were significantly increased in CLDN6-overexpressing cells. Furthermore, overexpression of CLDN6 enhanced cell-cell adhesion properties and attenuated sensitivity to anticancer drugs including doxorubicin, daunorubicin, and cisplatin. Taken together, the results indicate that aberrant expression of CLDN6 enhances malignant potentials and drug resistance of cervical ADC, possibly due to increased cell-cell adhesion properties and drug metabolism. Our findings provide an insight into a new therapeutic strategy, a CLDN6-targeting therapy, against cervical ADC.

Signet-ring cell/histiocytoid carcinoma in the axilla: A case report with genetic analysis using next-generation sequencing.

Signet-ring cell/histiocytoid carcinoma (SRCHC) is a very rare skin appendage cancer, with an extremely rare occurrence in the axilla. This study describes the 11th case of SRCHC occurring in the axilla and reports the first gene alteration analysis performed for SRCHC. An 85-year-old Japanese male presented with a tumor in the left axilla. Biopsy of the axilla nodule demonstrated diffuse proliferation of histiocytoid neoplastic cells and signet-ring cells in the dermis and subcutis. Immunohistochemistry revealed loss of E-cadherin expression in these neoplastic cells. Accordingly, SRCHC of the axilla was diagnosed. Genetic analysis using next-generation sequencing demonstrated missense mutation of PIK3CA (c1633G>A, pGlu545Lys) and no CDH1 gene mutation.SRCHC of the axilla is considered equivalent to a histiocytoid variant of invasive lobular breast carcinoma. The present SRCHC case demonstrated a pathogenic PIK3CA mutation, which is observed in invasive lobular carcinoma. Additional large case studies are required to clarify the clinicopathological features and gene alterations in SRCHC of the axilla.

Moesin affects the plasma membrane expression and the immune checkpoint function of CD47 in human ovarian clear cell carcinoma.

Among major histological subtypes of epithelial ovarian cancer, a higher incidence of ovarian clear cell carcinoma (OCCC) is observed in East Asian populations, particularly in Japan. Despite recent progress in the immune checkpoint inhibitors for a wide variety of cancer cell types, patients with OCCC exhibit considerably low response rates to these drugs. Hence, urgent efforts are needed to develop a novel immunotherapeutic approach for OCCC. CD47, a transmembrane protein, is overexpressed in almost all cancer cells and disrupts macrophage phagocytic activity in cancer cells. Ezrin-Radixin-Moesin (ERM) family member of proteins serve as scaffold proteins by crosslinking certain transmembrane proteins with the actin cytoskeleton, contributing to their plasma membrane localization. Here, we examined the role of ERM family in the plasma membrane localization and functionality of CD47 in OCCC cell lines derived from Japanese women. Confocal laser scanning microscopy analysis showed colocalization of CD47 with all three ERM in the plasma membrane of OCCC cells. RNA interference-mediated gene silencing of moesin, but not others, decreased the plasma membrane expression and immune checkpoint function of CD47, as determined by flow cytometry and in vitro phagocytosis assay using human macrophage-like cells, respectively. Interestingly, clinical database analysis indicated that moesin expression in OCCC was higher than that in other histological subtypes of ovarian cancers, and the expression of CD47 and moesin increased with the cancer stage. In conclusion, moesin is overexpressed in OCCC and may be the predominant scaffold protein responsible for CD47 plasma membrane localization and function in OCCC.

Intraductal papillary neoplasm of the bile duct with metachronous development in the downstream bile duct after radical resection.

We report a case of intraductal papillary neoplasms of the bile duct (IPNB) that metachronously developed twice in the downstream bile duct after radical resection. The first lesion was located in the left intrahepatic bile duct, the second lesion in the perihilar bile duct, and the third lesion in the distal bile duct. All lesions were IPNBs with associated invasive carcinoma (pancreatobiliary type). The depth of invasion was to the Glisson's capsule in the first lesion, to the subserosa in the second lesion, and to the fibromuscular layer in the third lesion, without lympho-vascular/perineural invasion and lymph-node metastasis. These were resected radically and had no biliary intraepithelial neoplasia and hyperplasia in the surrounding mucosa. In immunohistochemical examination, each lesion showed a different pattern. Although the downstream occurrence suggests intrabiliary dissemination, the mechanism of these metachronous developments may be multicentric. A literature review revealed that most metachronous cholangiocarcinomas have a grossly papillary appearance and tend to arise downstream. Our findings suggest that IPNB may develop metachronously in the residual bile duct after radical surgery, which may assist in early detection.

Porcine circoviruses in wild boars in Nagano Prefecture, Japan.

We investigated the prevalence of porcine circovirus 2 (PCV2) and 3 (PCV3) in wild boars captured or found dead in Nagano Prefecture in 2020. Based on PCR testing, 21 of 254 (8.3%) wild boars were PCV2-positive and 43 of 256 (16.8%) wild boars were PCV3-positive, 5 of 253 (2.0%) wild boars were both PCV2-positive and PCV3-positive. The frequency of detecting PCV3 in wild boars was significantly higher in adults than in juveniles (P=0.014). The PCV2-positive wild boars were found in all districts except for the North Alps and Hokushin, while PCV3-positive wild boars were found in all districts except for Suwa. This is the first report of PCV2 and PCV3 detected in wild boars in Japan.

Cellular Membrane Localization of Innate Immune Checkpoint Molecule CD47 Is Regulated by Radixin in Human Pancreatic Ductal Adenocarcinoma Cells.

In the past decade, immune checkpoint inhibitors have exhibited potent antitumor efficacy against multiple solid malignancies but limited efficacy against pancreatic ductal adenocarcinoma (PDAC). Cluster of differentiation (CD) 47, a member of the immunoglobulin G superfamily, is overexpressed in the surface membrane of PDAC and independently correlates with a worse clinical prognosis. Furthermore, CD47 functions as a dominant macrophage checkpoint, providing a potent "do not eat me" signal to enable cancer cells to evade the innate immune system. Thus, the blockade of CD47 is a promising immunotherapeutic strategy for PDAC. In this study, we determined whether ezrin/radixin/moesin (ERM) family members, which post-translationally modulate the cellular membrane localization of numerous transmembrane proteins by crosslinking with the actin cytoskeleton, contribute to the cellular membrane localization of CD47 in KP-2 cells derived from human PDAC. Immunofluorescence analysis showed that CD47 and ezrin/radixin were highly co-localized in the plasma membrane. Interestingly, gene silencing of radixin but not ezrin dramatically decreased the cell surface expression of CD47 but had little effects on its mRNA level. Furthermore, CD47 and radixin interacted with each other, as determined by a co-immunoprecipitation assay. In conclusion, radixin regulates the cellular membrane localization of CD47 as a scaffold protein in KP-2 cells.

Radixin modulates the plasma membrane localization of CD47 in human uterine cervical adenocarcinoma cells.

Despite the dramatic success of immune checkpoint blockers in treating numerous cancer cell types, current therapeutic modalities provide clinical benefits to a subset of patients with cervical cancers. CD47 is commonly overexpressed in a broad variety of cancer cells, correlates with poor clinical prognosis, and acts as a dominant macrophage checkpoint by interacting with receptors expressed on macrophages. It allows cancer cells to escape from the innate immune system and hence is a potential therapeutic target for developing novel macrophage checkpoint blockade immunotherapies. As the intracellular scaffold proteins, ezrin/radixin/moesin (ERM) family proteins post-translationally regulate the cellular membrane localization of numerous transmembrane proteins, by crosslinking them with the actin cytoskeleton. We demonstrated that radixin modulates the plasma membrane localization and functionality of CD47 in HeLa cells. Immunofluorescence analysis and co-immunoprecipitation assay using anti-CD47 antibody showed the colocalization of CD47 and all three ERM families in the plasma membrane, and the molecular interactions between CD47 and all three ERM. Interestingly, gene silencing of only radixin, reduced the CD47 plasma membrane localization and functionality by means of flow cytometry and phagocytosis assay but had little influence on its mRNA expression. Together, in HeLa cells radixin may function as a principal scaffold protein responsible for the CD47 plasma membrane localization.

Analytical Performance of a Novel Latex Turbidimetric Immunoassay, "Nanopia TARC", for TARC/CCL17 Measurement: A Retrospective Observational Study.

Thymus- and activation-regulated chemokine (TARC, also known as CCL17) is used as a biomarker for atopic dermatitis. The methods currently used for its measurement are complex, time-consuming, and require large machinery, warranting the need for a method that is simple, has a quick turnaround time, and requires less complex machinery. We evaluated the analytical performance of a novel latex turbidimetric immunoassay method, "Nanopia TARC", on 174 residual serum samples from patients with skin or allergic diseases. This evaluation included the assessment of the limit of blank/detection/quantification (LOB/D/Q), precision, accuracy, linearity, interference, and commutability between Nanopia TARC and "HISCL TARC", based on the chemiluminescent enzyme immunoassay (CLEIA) method. The LOB/D/Q values were 13, 57, and 141 pg/mL, respectively. The coefficient of variation of the repeatability was 0.9-3.8%, and that of the intermediate precision was 2.1-5.4%. The total error of the accuracy was 1.9-13.4%. The linearity was 141 and 19,804 pg/mL for TARC. The correlation coefficient between Nanopia TARC and HISCL TARC determined using the Passing-Bablok regression analysis was 0.999. Furthermore, the concordance of diagnostic criteria with AD was 92%. Nanopia TARC was confirmed to have the same analytical performance for TARC measurement as the existing CLEIA method.

Efficient direct membrane filtration (DMF) of municipal wastewater for carbon recovery: Application of a simple pretreatment and selection of an appropriate membrane pore size.

Considerable attention has been paid in recent years to the recovery and effective utilization of organic matter in municipal wastewater for the establishment of a circular economy. Direct membrane filtration (DMF) of municipal wastewater using microfiltration (MF) or ultrafiltration (UF) membranes to retain and concentrate the organic matter in municipal wastewater could be a practical option for this purpose. However, severe membrane fouling and high concentrations of organic matter remaining in the DMF permeate are concerns to be addressed. Application of a simple pretreatment using fixed biofilms was investigated to address these issues. In this study, experiments were carried out at an existing municipal wastewater treatment plant. A moving bed biofilm reactor (MBBR) process operated under a very short HRT of 1 h and DO concentration of 0.5 mg/L selectively degraded low-molecular-weight dissolved organic matter in municipal wastewater without degradation of membrane-recoverable suspended and colloidal organic matter. Application of the pretreatment did not reduce the amount of organic carbon recovered by DMF using an MF membrane (approximately 70% of the influent COD being recovered), while it dramatically mitigated the membrane fouling probably due to the alteration of characteristics of dissolved organic matter in wastewater. The pretreatment also reduced the concentration of organic matter in the DMF permeate by 41%: COD concentration in the DMF permeate was as low as 40 mg/L. With the established MBBR pretreatment, performances of MF (0.1 µm) and UF (molecular weight cut-off: 150,000) membranes for DMF were compared in parallel. It was found that the increase of the recoverable amount of organic matter by using UF was marginal (about 5%), whereas fouling in UF was much more severe than that in MF. The severe fouling in UF was caused by inorganic colloids such as FeS that could pass through MF membranes but be retained by UF membranes. Based on the results obtained in this study, it is concluded that MF is more suitable than UF for efficient DMF.

Enhancing the blood safety program in Myanmar: Report on projects of global extension of medical technologies of Japan.

The National Center for Global Health and Medicine has long collaborated with the blood program in Myanmar, and the Center started a new project in 2015 to enhance blood transfusion safety as part of a new set of projects of global extension of medical technologies that aims to improve public health and medicine in developing countries under public-private partnerships. The project resulted in remarkable achievements, including maintaining a high proportion of voluntary blood donations despite a rapidly growing demand for blood, ensuring blood safety from the donor to the recipient, and creating public-private partnerships. The project supported the introduction of blood grouping using the tube method at hospital blood banks, safety measures during blood transfusions, and effective use of blood products including component blood. The project identified the need for medical devices such as leukocyte filters, serofuges, and refrigerators to store blood products. The success of the project may depend on mutual understanding and trust based on the duration of collaboration, improvement of the requirement for medical safety (including blood safety) in the country, and shifting the mindset of partner companies in public-private partnerships to create new demand by encouraging improvement of the quality of care and requiring the safety of medical care. In this era of sustainable development goals, the hopes are that these experiences will help other countries seeking to improve their public health through public-private partnerships.

Successful measures to prevent the spread of bovine papular stomatitis in a dairy farm.

Nasal papules and oral ulcers were observed in calves that were group-housed at a dairy farm. The calves were diagnosed with bovine papular stomatitis (BPS) due to parapoxvirus (PPV) infection based on virologic examinations using polymerase chain reaction to detect PPV. To prevent the spread of BPS, we isolated the affected calves, made procedural changes so that the affected herd was managed after the healthy herd, disinfected the bedding with slaked lime, disinfected the stalls and fences with invert soap, and changed the animals' feed to soft grass which does not damage the oral cavity. As a result, we succeeded in control the infection quickly.

[Combination of percutaneous balloon angioplasty and aggressive medical intervention improves symptomatic basilar artery stenosis with a tortuous access route: case report].

UNLABELLED: The prognosis of symptomatic intracranial artery stenosis (SIAS) by medical treatment is poor. Percutaneous balloon angioplasty (PTA) or stenting is effective for SIAS. However, recently, aggressive medical intervention (AMI) has progressed and some drugs have been reported to improve stenosis or prevent the progression of stenosis. We describe a case where combination of PTA and the AMI improved symptomatic basilar artery stenosis with a tortuous access route. CASE: A 78-year-old man was admitted to our hospital suffering from acute brain infarction due to severe basilar artery stenosis. The AMI including cilostazol, statin, and eicosapentaenoic acid failed to prevent recurrence of the brain infarction. We performed PTA and 45% of the residual stenosis remained. We continued the AMI and the stenosis improved to 30% after 3 months. CONCLUSION: PTA with minimal risk should be considered for SIAS resistant to AMI. Even if residual stenosis remains, continuation of AMI may prevent recurrence of a brain infarction. The strategy of a combination of PTA with minimal risk and AMI may result in a better prognosis for SIAS.

Oxidative Kinetic Resolution of Acyclic Amines Based on Equilibrium Control.

An oxidative kinetic resolution of racemic acyclic amines was developed using an imine derivative as the resolving reagent and chiral phosphoric acid as the catalyst to give enantiomers in good yields with high to excellent enantioselectivities. The key to success of the title reaction was the equilibrium control by adjusting the ratio of the resolving reagent, and unique enantiodivergence was observed depending on the equilibrium displacement.

Characteristics of physical prefrailty among Japanese healthy older adults.

AIM: The purpose of the present study was to clarify the characteristics of frailty at an early stage (prefrailty) in a healthy elderly Japanese population. METHODS: The participants were 620 healthy older adults (age range 60-89 years) who were current students or graduates of a community college for older adults in Japan. All participants were evaluated using the Kihon Checklist, a tool developed to screen for frailty in Japan. The participants were categorized by the Fried criteria (lost weight, handgrip strength, walking speed, exhaustion, physical activity) into either a prefrailty (1-2 criteria) or a non-frailty (0 criterion) group. RESULTS: In the logistic regression model for the prevalence of prefrailty, significant and independent determinants were chronic constipation (odds ratio [OR] 2.1, 95% confidence interval [CI] 1.15-3.90, P = 0.016), occurrence of incontinence (OR 2.0, 95% CI 1.39-2.82, P < 0.001), unable to climb stairs (OR 3.0, 95% CI 1.26-7.02, P = 0.013), dry mouth (OR 1.5, 95% CI 1.04-2.22, P = 0.029), a lack of fulfillment (OR 3.2, 95% CI 1.26-7.93, P = 0.015), found easy tasks difficult (OR 2.8, 95% CI 1.30-5.85, P = 0.008) and felt helpless (OR 2.1, 95% CI 1.02-4.39, P = 0.044). CONCLUSIONS: These results suggest that the appearance of autonomic failure, oral malfunction and some psychological factors in relation to depressed mood, but not anthropometric measurements, are characteristic of prefrailty. Geriatr Gerontol Int 2017; 17: 1568-1574.

Involvement of substance P in the development of cisplatin-induced acute and delayed pica in rats.

BACKGROUND AND PURPOSE: Although substance P (SP) and neurokinin NK1 receptors have been reported to be involved in cisplatin-induced acute and delayed emesis, their precise roles remain unclear. Pica, the consumption of non-nutrient materials such as kaolin in rats, can be used as a model of nausea in humans. We investigated the time-dependent changes in cisplatin-induced pica and the involvement of SP and NK1 receptors in this behaviour. EXPERIMENTAL APPROACH: Rats were administered cisplatin with or without a daily injection of a 5-HT3 receptor antagonist (granisetron) or an NK1 receptor antagonist (aprepitant), and kaolin intake was then monitored for 5 days. The effects of granisetron on the cisplatin-induced expression of preprotachykinin-A (PPT-A) mRNA, which encodes mainly for SP, and on SP release in the medulla, measured by in vivo brain microdialysis, were also investigated. KEY RESULTS: Cisplatin induced pica within 8 h of its administration that continued for 5 days. Granisetron inhibited the acute phase (day 1), but not the delayed phase (days 2-5), of pica, whereas aprepitant abolished both phases. Within 24 h of the injection of cisplatin, PPT-A mRNA expression and SP release in the medulla were significantly increased; these findings lasted during the observation period and were inhibited by granisetron for up to 24 h. CONCLUSIONS AND IMPLICATIONS: The profiles of cisplatin-induced pica in rats are similar to clinical findings for cisplatin-induced emesis in humans, and we showed that SP production in the medulla and activation of NK1 receptors are involved in this cisplatin-induced pica.

Involvement of hypothalamic cyclooxygenase-2, interleukin-1ß and melanocortin in the development of docetaxel-induced anorexia in rats.

Docetaxel, a taxane derivative, is frequently used for the treatment of advanced breast cancer, non-small cell lung cancer, and metastatic prostate cancer. Clinical reports demonstrated that docetaxel-based chemotherapy often induces anorexia, but the etiology is not completely understood. To elucidate possible mechanisms, we investigated the involvement of central interleukin (IL)-1ß, cyclooxygenase (COX)-2, and pro-opiomelanocortin (POMC) in the development of docetaxel-induced anorexia in rats. Rats received docetaxel (10mg/kg, i.p.) with or without pretreatment with selective COX-2 inhibitors, NS-398 (10 and 30 mg/kg, i.g.) or celecoxib (10 and 30 mg/kg, i.g.), and a non-selective COX inhibitor, indomethacin (10mg/kg, i.g.), then food intake was monitored for 24h after administration. We also examined expression of IL-1ß, COX-2, and POMC mRNA in hypothalamus of docetaxel-treated rats and the effect of a COX-2 inhibitor on docetaxel-induced POMC mRNA expression. Food consumption in rats was significantly decreased 24h after administration of docetaxel and anorexia was partially reversed by all COX inhibitors. Administration of docetaxel increased IL-1ß, COX-2, and POMC mRNA expression in the hypothalamus of rats. The time required to increase these gene expressions was comparable to the latency period of docetaxel-induced anorexia in rats. In addition, pretreatment with COX-2 inhibitors suppressed docetaxel-induced expression of POMC mRNA. These results suggest that IL-1ß and COX-2 mRNA expression and subsequent activation of POMC in the hypothalamus may contribute to the development of docetaxel-induced anorexia in rats.