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Abstract Introduction Mutated human epidermal growth factor receptor 2 (HER2) is an oncogene with critical pathogenic roles in breast cancer. HER2-low-positive breast cancer is a recently described subtype. We aimed to explore the clinical and molecular characteristics of gastric cancer with low HER2 expression, drawing on recent developments in breast cancer subtypes. Materials and methods This retrospective study involved 129 patients with HER2-non-amplified gastric cancer treated in Iwate prefectural Iwai Hospital from 2013 to 2019. Tumors were classified as HER2-null or low-positive based on immunohistochemistry score 0 or 1 + or 2 + with HER2 negativity in situ hybridization, respectively. Statistical analyses, including Kaplan–Meier analyses and Cox proportional hazards model were conducted. Results Low HER2 expression was present in 26% (33/129) of the patients. Clinicopathological characteristics were not significantly different between the HER2-low and null groups. Kaplan–Meier analysis of overall survival was significantly longer in the HER2-low group than in the HER2-null group (P = 0.01). In multivariate Cox regression analysis, HER2-null status was associated with worse survival (hazard ratio 3.01; 95% confidence interval 1.18–7.65; and P = 0.02). Conclusion This study highlights the prognostic importance of low HER2 expression in gastric cancer, similar to that observed in HER2-low-positive breast cancer, and suggests reclassification of gastric cancer to improve personalized treatment. Future studies should elucidate the molecular underpinnings of low HER2 expression in gastric cancer to guide novel therapeutic strategies and improve outcomes.

Abstract The current study aimed to identify the indications for manual reduction in incarcerated obturator hernias (OH). Further, whether time to symptom onset and manual reduction outcomes can be predictors of bowel viability and the need for bowel resection in incarcerated OH were examined. This retrospective study included 26 patients with incarcerated OH who underwent surgery. All patients underwent manual reduction, and computed tomography scan after manual reduction confirmed hernia release. Multivariate analyses were performed to determine the predictors of bowel resection. The bowel resection group had a significantly longer average time to symptom onset than the nonbowel resection group (88 vs 36 h). Further, the bowel resection group was more likely to have failed manual reduction than the nonbowel resection group. A time to symptom onset of ≥ 72 h and failed manual reduction were significant predictors of bowel viability. Age, sex, hernia localization, American Society of Anesthesiologists physical status score, and laboratory findings did not differ significantly between the bowel resection and nonbowel resection groups. Time to symptom onset and manual reduction outcomes are significant predictors of bowel viability in incarcerated OH. Patients with a time to symptom onset of ≥ 72 h and failed manual reduction require surgical evaluation due to a high risk of bowel nonviability. Therefore, a cautious approach is required in the management of OH, and further research on optimized treatment protocols should be conducted.

The area offshore of Minamitorishima Island, Northwestern Pacific Ocean, contains large amounts of seafloor mineral resources such as ferromanganese (Fe–Mn) nodules, Fe–Mn crusts, and rare-earth element and yttrium (REY)-rich muds. In this study, we applied stratigraphic Os isotopic dating to a Fe–Mn nodule for the first time to date its formation/depositional age, and mineralogical and texturally characterized a complementary nodule. Based on macroscopic and microscopic observations, the studied Fe–Mn nodules can be divided into three layers: Layers L2, L1, and L0 from core to rim. Under the microscope, the Fe–Mn nodules are dominated by vernadite and Fe-oxyhydroxide. In particular, Layer L1 is dominated by banded-columnar vernadite and contains lower amounts of clay minerals derived from the detrital component than Layers L2 and L0. The bulk major and trace element geochemical compositions of sampled layers in the Fe–Mn nodules all plotted in the hydrogenous field in several discrimination diagrams. The Os isotopic ages determined by fitting to the paleo-seawater 187Os/188Os curve can be divided into three clusters (35.7–31.0, 19.0–9.0, and 4.0–1.0 Ma, corresponding to Layers L2, L1, and L0, respectively). These Os isotopic ages indicate that two periods of very slow growth or growth hiatuses occurred during the formation of the Fe–Mn nodule; these age gaps are related to the intermittent (discontinuous) timings of the beginning of Fe–Mn nodule formation offshore Minamitorishima Island.

Abstract Emergent surgery is a common approach for incarcerated obturator hernias, with high morbidity and mortality rates. Moreover, there have been reports of elective surgery cases after noninvasive manual reduction. For a decade, the initial approach in our institution is also manual reduction unless bowel viability is poor. This study aimed to clarify the efficacy and safety of manual reduction followed by elective surgery. We retrospectively reviewed 50 cases of incarcerated obturator hernia from 2010 to 2022 at Iwate Prefectural Iwai Hospital. Manual reduction was attempted in 31 (62%) patients. The reduction was successful in 21 (42%) patients, and most of them received mesh repair using the extraperitoneal approach as elective surgery. However, two patients underwent emergent surgery in the waiting period because of late-onset constriction and a small bowel perforation. Patients with irreducible hernia underwent emergent surgery, except for two patients who received the best supportive care. Postoperative complications were observed in 5% and 22% of reducible and irreducible cases, respectively. Postoperative mortality was zero in both groups. Manual reduction is useful in some cases, but careful observation is needed because late-onset constriction and perforation could occur.

Abstract Background Laparoscopic surgery has reduced surgical morbidity and postoperative duration of hospital stay. Gas embolism is commonly known as a risk factor for all laparoscopic procedures. We report a case of severe cerebral infarction presumably caused by paradoxical CO2 embolism in laparoscopic partial hepatectomy with an insufflation management system. Case presentation A male in his 60 s was diagnosed with recurrence of liver metastasis in the right hepatic lobe after laparoscopic lower anterior resection for rectal cancer. We performed laparoscopic partial hepatectomy with an AirSeal® under 10 mmHg of intra-abdominal pressure. During the surgery, the patient’s end-tidal CO2 and percutaneous oxygen saturation dropped from approximately 40–20 mmHg and 100–90%, respectively, while the heart rate increased from 60 to 120 beats/min; his blood pressure remained stable. Postoperatively, the patient developed right hemiplegia and aphasia. Brain magnetic resonance imaging showed cerebral infarction in the broad area of the left cerebral cortex. Thereafter, transesophageal echocardiography revealed a patent foramen ovale, suggesting cerebral infarction due to paradoxical gas embolism. Conclusions A patent foramen ovale is found in approximately 15–20% of healthy individuals. While gas embolism is a rare complication of laparoscopic surgery, cerebral infarction must be considered a possible complication even if the intra-abdominal pressure is constant under 10 mmHg with an insufflation management system.

Dendritic cell immunoreceptor (DCIR) is an inhibitory C-type lectin receptor that acts as a negative regulator in the immune system and bone metabolism. We previously revealed that DCIR deficiency enhanced osteoclastogenesis and antigen presentation of dendritic cells, and that asialo-biantennary N-glycan (NA2) functions as a ligand for DCIR. NA2 binding to DCIR suppressed murine and human osteoclastogenesis that occurs in the presence of M-CSF and RANKL. The DCIR-NA2 axis, therefore, plays an important role in regulating osteoclastogenesis in both mice and humans, although the underlying mechanisms remain unclear. Here we found that Dcir−/− bone marrow–derived macrophages (BMMs) exhibited greater proliferative and differentiation responses to M-CSF and RANKL, respectively, than wild-type (WT) BMMs. Moreover, Dcir−/− osteoclasts (OCs) increased resorptive activity and cell fusion more significantly than WT OCs. DCIR deficiency affects gene expression patterns in OCs, and we found that the expression of neuraminidase 4 was increased in Dcir−/− OCs. Furthermore, DCIR-NA2 interaction in WT BMMs, but not Dcir−/− BMMs, decreased Akt phosphorylation in response to M-CSF and RANKL. These data suggest that DCIR regulates osteoclastogenesis by downregulating M-CSF and RANKL signaling, and that DCIR-mediated signaling may contribute to the terminal modification of oligosaccharides by controlling the expression of glycosylation enzymes.

Objective: Adhesion barriers have not traditionally been used during laparoscopic cholecystectomy (lap-chole), and so there are no data confirming the safety of the dextrin hydrogel adhesion barrier (AdSpray®) for the use during lap-chole. The aim of this prospective study is to investigate the safety of AdSpray® for elective lap-chole. Methods: Between July 2019 and March 2021, we conducted a prospective study at Saiseikai Kumamoto Hospital to assess the safety of applying AdSpray® around the hepatoduodenal ligament, liver surface, and port area after extraction of the gallbladder during elective lap-chole. We assessed patient morbidity, mortality, and laboratory data on postoperative day 1 and at the outpatient follow-up visit. Results: A total of 100 patients (43 men, 57 women) received AdSpray® application during elective lap-chole at our institution. The mean patient age was 56.5 ± 14.8 y, and the mean body mass index was 25.1 ± 4.8 kg/m2. Preoperative gallbladder drainage was performed in 3 (3.0%) patients as conservative treatment for acute cholecystitis. The mean operative time was 91.7 ± 35.1 min, and the median blood loss was 5 mL (range, 5–120 mL). Intraoperative gallbladder perforation was observed in 4.0% of patients (n = 4). There was no intraoperative bile duct injury. One (1.0%) patient had a postoperative subhepatic fluid collection without bacterial infection. The mean white blood cell count and C-reactive protein level was significantly elevated on postoperative day 1 but returned to preoperative levels by the time of the follow-up visit. All patients were successfully discharged after surgery, and the median postoperative hospital stay was 2 d (range, 2–9 d). Conclusion: Applying AdSpray® during elective lap-chole is safe, with an acceptable rate of postoperative complications.

Abstract Surface ion traps are among the most promising technologies for scaling up quantum computing machines, but their complicated multi-electrode geometry can make some tasks, including compensation for stray electric fields, challenging both at the level of modeling and of practical implementation. Here we demonstrate the compensation of stray electric fields using a gradient descent algorithm and a machine learning technique, which trained a deep learning network. We show automated dynamical compensation tested against induced electric charging from UV laser light hitting the chip trap surface. The results show improvement in compensation using gradient descent and the machine learner over manual compensation. This improvement is inferred from an increase of the fluorescence rate of 78% and 96% respectively, for a trapped $$^{171}$$ 171 Yb $$^+$$ + ion driven by a laser tuned to $$-7.8$$ - 7.8 MHz of the $$^2$$ 2 S $$_{1/2}\leftrightarrow ^2$$ 1 / 2 ↔ 2 P $$_{1/2}$$ 1 / 2 Doppler cooling transition at 369.5 nm.

Abstract 2p15p16.1 microdeletion syndrome is a recently recognized congenital disorder characterized by developmental delay and dysmorphic features. RP2-associated retinal disorder (RP2-RD) is an X-linked inherited retinal disease with a childhood onset caused by a loss-of-function variant in the RP2 gene. Here, we describe a 14-year-old boy with double diagnoses of 2p15p16.1 microdeletion syndrome and RP2-RD. The recurrence risk of each condition and the indication for potential therapeutic options for RP2-RD are discussed.

Abstract Hydrogen isotopes have been widely used as powerful tracers to understand the origin of terrestrial water and the water circulation between the surface and the deep interior of the Earth. However, further quantitative understanding is hindered due to a lack of observations about the changes in D/H ratios of a slab during subduction. Here, we report hydrogen isotope data of olivine-hosted melt inclusions from active volcanoes with variable depths (90‒550 km) to the subducting Pacific slab. The results show that the D/H ratio of the slab fluid at the volcanic front is lower than that of the slab fluid just behind the volcanic front. This demonstrates that fluids with different D/H ratios were released from the crust and the underlying peridotite portions of the slab around the volcanic front. The results also show that the D/H ratios of slab fluids do not change significantly with slab depths from 300 to 550 km, which demonstrates that slab dehydration did not occur significantly beyond the arc. Our estimated δD‰ value for the slab materials that accumulated in the mantle transition zone is > − 90‰, a value which is significantly higher than previous estimates.

Abstract This 10-year retrospective observational study investigated longitudinal losses in psoas major and paraspinal muscle area in 1849 healthy individuals (1690 male, 159 female) screened using computed tomography. Logistic regression analysis revealed significant decreases in psoas major and paraspinal muscle area at 10 years relative to the baseline area regardless of age or sex, starting at 30 years of age. Only aging [≥ 50 s (odds ratio [OR]: 1.72; 95% confidence interval [CI] 1.05–2.84; p = 0.03) and ≥ 60 s (OR: 2.67; 95% CI 1.55–4.60; p 45 min daily (Calories = METs (walking: 3.0) × duration of time (h) × weight (60 kg) × 1.05) can reduce paraspinal muscle loss, which may in turn decrease the risk of falls, low-back pain, and sarcopenia.

Abstract Background Subtotal cholecystectomy in patients with severe acute cholecystitis is considered a “bailout” option when the safety of the bile duct cannot be guaranteed. However, subtotal cholecystectomy has a long-term risk of remnant cholecystitis. The appropriate management of remnant cholecystitis has not been fully elucidated. Case presentation Case 1 was a 66-year-old man who had undergone subtotal cholecystectomy 14 years prior to the development of remnant cholecystitis. We first performed endoscopic gallbladder drainage to minimize inflammation, and then proceeded with elective surgery. We performed a reconstituting procedure for the residual gallbladder due to significant adhesions between the cystic and common bile ducts. Case 2 was a 56-year-old man who had undergone subtotal cholecystectomy for abscess-forming perforated cholecystitis 2 years prior to the development of remnant cholecystitis. He underwent endoscopic drainage followed by complete remnant cholecystectomy 4 months later. Conclusion Endoscopic gallbladder drainage is a useful strategy to improve inflammation and reduce the risk of bile duct injury during remnant cholecystectomy.

Abstract Aim Severe postoperative pleural effusion (sPOPE) after hepatectomy can lead to respiratory distress and may require thoracic drainage, leading to prolonged hospitalization. Preventive chest tube insertion may be useful for patients at high risk for sPOPE. We aimed to develop a predictive model for sPOPE after hepatectomy and evaluate indications for preventive chest tube insertion using our model. Methods We evaluated all patients who underwent hepatectomy from 2013 to 2020. Risk factors for sPOPE were used to develop a predictive model for sPOPE, which was validated in a cohort that received preventative chest tube placement postoperatively. Results A total of 325 patients were analyzed. Thirty‐one (9.5%) patients had a preventive chest tube placed at the end of their operation. Twenty‐one patients out of the remaining 294 patients developed sPOPE. Multivariate analysis identified resection containing segment 8 [relative risk (RR) 3.24, P = .022], intraoperative bleeding ≥ 500 g (RR 4.02, P = .008), intraoperative diaphragmatic incision (RR 6.96, P = .042) and open hepatectomy (RR 7.51, P = .016) as independently associated with sPOPE. The estimated probability of sPOPE ranged from 0.4% in patients with none of these factors to 73.4% in the presence of all factors. Among the 31 patients who received a preventive chest tube, more patients in the high‐risk group defined by the model had postoperative pleural effusions compared to the low‐risk group (P = .012). Conclusion Our predictive model for sPOPE using four risk factors allows for reliable prediction and may be useful for selection of preventive chest tube in patients undergoing hepatectomy.

TARM1 is a LILR family member that drives cell signalling via interactions with FcRγ. Here the authors show that TARM1 binds collagens to activate dendritic cells and thereby is an effector of inflammatory arthritis, plus provide a soluble TARM-Fc fusion protein that can limit collagen-induced arthritis in mice.

Abstract Introduction Our previous clinical studies have demonstrated the short-term efficacy and safety of the sirolimus gel for patients with tuberous sclerosis complex (TSC). However, long-term clinical evidence is lacking. Our objective was to assess the safety and efficacy of long-term treatment with the sirolimus gel for the skin lesions of TSC patients. Methods We conducted a multicenter, open-label, uncontrolled clinical trial in 94 Japanese patients with TSC. Patients applied the 0.2% sirolimus gel on their face or head twice daily for > 52 weeks (maximum 136 weeks for safety). The safety endpoints were the rate of adverse event (AE)-caused discontinuation (primary endpoint) and the incidence of AEs. The efficacy endpoint was the response rate of angiofibromas, cephalic plaques, and hypomelanotic macules. Results Among 94 enrolled patients (mean age, 21 years; range 3–53 years), the rate of AE-caused discontinuation was 2.1% (2/94 patients). Although application site irritation and dry skin occurred relatively frequently, none of the drug-related AEs were serious; most of the drug-related AEs resolved rapidly. The major drug-related AEs (≥ 5% in incidence) were application site irritation (30.9%), dry skin (27.7%), acne (20.2%), eye irritation (8.5%), pruritus (8.5%), erythema (7.4%), dermatitis acneiform (6.4%), and dermatitis contact (5.3%). The response rates of angiofibromas, cephalic plaques, and hypomelanotic macules were 78.2% [95% confidence interval (CI) 68.0–86.3%], 66.7% (95% CI 51.1–80.0%), and 72.2% (95% CI 46.5–90.3%), respectively. Conclusions The gel was well tolerated for a long time by patients with TSC involving facial skin lesions and continued to be effective. Trial Registration ClinicalTrials.gov identifier: NCT02634931.

Abstract Subduction‐related lavas have higher Fe3+/∑Fe than midocean ridge basalts (MORB). Hypotheses for this offset include imprint from subducting slabs and differentiation in thickened crust. These ideas are readily tested through examination of the time‐dependent evolution of slab‐derived signatures, thickening crust of the overriding plate, and evolving redox during subduction initiation. Here, we present Fe3+/ΣFe and volatile element abundances of volcanic glasses recovered from International Ocean Discovery Program (IODP) Expedition 352 to the Izu‐Bonin‐Mariana (IBM) forearc. The samples include forearc basalts (FAB) that are stratigraphically overlain by low‐ and high‐silica boninite lavas. The FAB glasses have 0.18–0.85 wt% H2O, 75–233 ppm CO2, S contents controlled by saturation with a sulfide phase (602–1,386 ppm), Ba/La from 3.9‐10, and Fe3+/ΣFe ratios from 0.136 to 0.177. These compositions are similar to MORB and suggest that decompression melting of dry and reduced mantle dominates the earliest stages of subduction initiation. Low‐ and high‐silica boninite glasses have 1.51–3.19 wt% H2O, CO2 below detection, S contents below those required for sulfide saturation (5–235 ppm), Ba/La from 11 to 29, and Fe3+/∑Fe from 0.181 to 0.225. The compositions are broadly similar to modern arc lavas in the IBM arc. These data demonstrate that the establishment of fluid‐fluxed melting of the mantle, which occurs in just 0.6–1.2 my after subduction initiation, is synchronous with the production of oxidized, mantle‐derived magmas. The coherence of high Fe3+/∑Fe and Ba/La ratios with high H2O contents in Expedition 352 glasses and the modern IBM arc rocks strongly links the production of oxidized arc magmas to signatures of slab dehydration.

Abstract Background Human 15q11–13 is responsible for Prader-Willi syndrome (PWS) and Angelman syndrome (AS) and includes several imprinted genes together with bipartite elements named AS-IC (imprinting center) and PWS-IC. These concertedly confer allele specificity on 15q11–13. Here, we report DNA methylation status of 15q11–13 and other autosomal imprinted differentially methylated regions (iDMRs) in cases with various deletions within the PWS/AS-responsible region. Methods We performed array-based methylation analysis and examined the methylation status of CpG sites in 15q11–13 and in 71 iDMRs in six cases with various microdeletions, eight cases with conventional deletions within 15q11–13, and healthy controls. Results We detected 89 CpGs in 15q11–13 showing significant methylation changes in our cases. Of them, 14 CpGs in the SNORD116s cluster presented slight hypomethylation in the PWS cases and hypermethylation in the AS cases. No iDMRs at regions other than 15q11–13 showed abnormal methylation. Conclusions We identified CpG sites and regions in which methylation status is regulated by AS-IC and PWS-IC. This result indicated that each IC had unique functions and coordinately regulated the DNA methylation of respective alleles. In addition, only aberrant methylation at iDMRs in 15q11–13 leads to the development of the phenotypes in our cases.

Volatile exchange between the Earth’s interior and surface layers is one of the central issues in mantle geochemistry. Here the authors present evidence that chlorine is transferred from the surface to the deep mantle by subducted oceanic crust, forming a chlorine-rich mantle reservoir.

Abstract International Ocean Discovery Program Expedition 352 to the Izu‐Bonin forearc cored over 800 m of basement comprising boninite and boninite‐series lavas. This is the most extensive, well‐constrained suite of boninite series lavas ever obtained from in situ oceanic crust. The boninites are characterized as high‐silica boninite (HSB), low‐silica boninite (LSB), or basaltic boninite based on their SiO2‐MgO‐TiO2 relations. The principal fractionation products of all three series are high‐Mg andesites (HMA). Lavas recovered >250 meters below seafloor (mbsf) erupted at a forearc spreading axis and are dominated by LSB and HMA. Lavas recovered from

Abstract International Ocean Discovery Program Expedition 352 to the Bonin forearc drilled the sequence of volcanic rocks erupted in the immediate aftermath of subduction initiation along the western margin of the Pacific Plate. Pristine volcanic glasses collected during this expedition were analyzed for major and trace elements, halogens, sulfur, and H and O isotopes with goals of characterizing the fluids and melts of subducted materials that were involved in generating the nascent upper plate crust. Incompatible trace element compositions of the oldest lavas (forearc basalts [FAB]) are similar to those of the most depleted mid‐ocean ridge basalts globally. Most FAB were generated by decompression melting during seafloor spreading in a near‐trench, supra‐subduction zone environment with only minor involvement of diverse and generally dilute water‐rich fluids from the subducting plate. Boninite series glasses are enriched in incompatible trace elements mobilized from the subducting plate, but strongly depleted in other elements, such as the middle‐heavy rare‐earth elements. These traits are attributed to generation of boninites largely by flux melting involving water‐rich melts first derived from the leading edge of subducted basaltic crust and then from both subducted crust and sediment. These melts were generated at low pressures as the shallow, embryonic slab extracted heat from hot asthenosphere near the trench. The progressive depletion of the mantle source for the FAB‐through‐boninite sequence suggests that the asthenospheric mantle remained trapped above the nascent subducting plate for the first several million years of subduction beneath the Philippine Sea Plate.

Abstract Background Deafness, dystonia, and cerebral hypomyelination (DDCH) is an X‐linked disorder due to hemizygous mutations of BCAP31. Methods We report an 8‐year‐old boy with DDCH who possibly accompanied mitochondrial dysfunction. Clinical evaluation, respiratory chain enzyme assay, and whole exome sequencing analysis were performed. Results Mitochondrial dysfunction was suspected by respiratory chain enzyme assay on his cultured skin fibroblasts which showed significantly decreased complex I enzyme activity. Whole exome sequencing analysis revealed a recurrent BCAP31 mutation (c.97C>T:p.Gln33*) which confirmed the diagnosis of DDCH for the patient. Conclusion We speculate that mitochondrial dysfunction may be a feature in patients with DDCH.

In adult liver transplantation, renoportal anastomosis (RPA) has been introduced as a useful technique for patients with grade 4 portal vein thrombosis and a splenorenal shunt. Here, we report a pediatric case in which RPA allowed a left lateral lobe living donor liver transplantation (LDLT) despite portal vein thrombosis and a large splenorenal shunt. At 36 days old, the patient underwent a Kasai operation for biliary atresia. At 17 months old, she underwent LDLT because of repetitive cholangitis. Pretransplant examinations revealed a large splenorenal shunt and portal vein thrombosis. Simple end-to-end portal reconstruction and clamping of the collateral route after removing the thrombosis were unsuccessful. Thus, RPA was performed using a donor superficial femoral vein as an interpositional graft. The portal vein pressure was 20 mm Hg after arterial reperfusion. Ligation of the splenic artery reduced the portal vein pressure. Although she developed severe acute cellular rejection and chylous ascites, there were no signs of portal vein complications. She was discharged 73 days after transplantation without any signs of renal dysfunction. The patient’s condition was good at her last follow-up, 22 months after transplantation. To our knowledge, this is the youngest case of RPA in pediatric left lateral lobe LDLT. Additionally, this is the first case of RPA with splenic artery ligation and using the donor’s superficial femoral vein as the venous graft for RPA. Although long-term follow-up is necessary, RPA could be a salvage option in LDLT in infants if other methods are unsuccessful.

Abstract Skeletal muscle metastasis from pancreatic cancer is rare. We present a 72-year-old female patient with unresectable pancreatic tail cancer. Fifteen months after the introduction of the chemoradiotherapy, an 18-mm elastic hard tumor was found in her right chest wall and resected after confirmation of no other metastatic lesions. Postoperative pathological examination diagnosed it as a muscle metastasis from the pancreatic cancer, and the patient has since been continuing chemotherapy for 10 months. A review of the literature regarding skeletal muscle metastasis from pancreatic cancer is also presented.

Cancer-immunotherapy targeting programmed cell death 1 (PD-1) activates tumor-specific T cells and provides clinical benefits in various cancers. However, the molecular basis of PD-1 function is still enigmatic. Especially, it is unclear which signaling pathway PD-1 primarily targets. Besides, the capacity of PD-1 to inhibit the T cell receptor (TCR)-dependent activation of T cells in the presence of co-stimulation is also controversial. Here we used co-culture systems of T cells and antigen-presenting cells with targeted deletion and overexpression of co-receptors and ligands and examined the inhibitory potency of PD-1 against T cell activation upon TCR stimulation with CD28 and ICOS co-stimulation. As an unambiguous criterion of T cell activation, we used the acquisition of cytokine production capacity, which represents one of the most important functions of T cells. PD-1 inhibited functional T cell activation upon TCR stimulation in the absence as well as in the presence of CD28 co-stimulation, indicating that PD-1 can directly inhibit TCR signal. Notably, CD28 co-stimulation rather attenuated the efficiency of PD-1 in inhibiting TCR-dependent functional T cell activation. In addition, PD-1 inhibited TCR-dependent functional T cell activation with ICOS co-stimulation as efficiently as that with CD28 co-stimulation. Furthermore, we found that the maintenance of antigen-induced follicular helper T (TFH) cells that required ICOS co-stimulation was persistently restrained by PD-1 in vivo. These findings indicate that PD-1 primarily targets TCR signal in the inhibition of functional T cell activation. Thus, PD-1 functions as the rheostat of T cell activation rather than an inhibitor of a specific stimulatory co-receptor.

Toxoplasma gondii is a major protozoan parasite and infects human and many other warm-blooded animals. The infection leads to Toxoplasmosis, a serious issue in AIDS patients, organ transplant recipients and pregnant women. Neospora caninum, another type of protozoa, is closely related to Toxoplasma gondii. Infections of the protozoa in animals also causes serious diseases such as Encephalomyelitis and Myositis-Polyradiculitis in dogs or abortion in cows. Both Toxoplasma gondii and Neospora caninum have similar nucleoside triphosphate hydrolases (NTPase), NcNTPase and TgNTPase-I in Neospora caninum and Toxoplasma gondii, respectively. These possibly play important roles in propagation and survival. Thus, we targeted the enzymes for drug discovery and tried to establish a novel high-standard assay by a combination of original biochemical enzyme assay and fluorescent assay to determine ADP content. We then validated whether or not it can be applied to high-throughput screening (HTS). Then, it fulfilled criterion to carry out HTS in both of the enzymes. In order to identify small molecules having inhibitory effects on the protozoan enzyme, we also performed HTS using two synthetic compound libraries and an extract library derived from marine bacteria and then, identified 19 compounds and 6 extracts. Nagasaki University collected many extracts from over 18,000 marine bacteria found in local Omura bay, and continues to compile an extensive collection of synthetic compounds from numerous drug libraries established by Japanese chemists.

Very recently, the immunotherapies against cancer, autoimmune diseases, and infection have been feasible and promising. Thus, we have examined the possibility whether or not human gamma delta T cells can be applied for the novel immunotherapies. We previously established the cells stably maintaining NFkB-driven human secreted embryonic alkaline phosphatase (SEAP) expression. The cells can be used to determine the transcription activity of NFkB with high-standard dynamic range and accuracy. Because IL-18 is a kind of cytokines that enhances cytotoxicity and activity of human gamma delta T cells through NFkB activation, we have focused on the activity and signaling of IL-18. In this study, we modified the previous reporter cell that can determine the transcription activity of NFkB to express two subunits consisted of human IL-18 receptor. The modified cells secreted SEAP in response to treatment with human recombinant IL-18 in a concentration-dependent manner. We also observed the concentration-dependently enhancement of NFkB activity in the cells treated with mouse recombinant IL-18 although the affinity was lower compared to human recombinant IL-18. We also previously established the cells stably expressing and secreting human recombinant IL-18 and then validated whether or not the conditioned medium from the cells activate NFkB transcription activity using this assay. Our university has kept collecting many extracts from over 18,000 marine bacteria in our local sea around Omura bay—fungi, plants for Chinese herbal medicine, and so on—and also have kept gathering synthetic compounds from many Japanese chemists as drug libraries. Finally, in order to identify drugs mimicking IL-18 biological activity or possessing inhibitory effects on IL-18-induced NFkB, we demonstrated drug screening using number of extracts derived from marine bacteria and synthetic compounds.

Expression of ABCA1 is regulated by transcription of the gene and calpain-mediated proteolytic degradation, and inhibition ABCA1 degradation results in increased ABCA1 and HDL biogenesis in vitro. We examined whether this approach could be a potential antiatherogenic treatment. Although probucol inhibits both the activity and degradation of ABCA1, its oxidized products, spiroquinone and diphenoquinone, reduce degradation of ABCA1 without inhibiting its activity or altering transcription of the ABCA1 gene. Accordingly, both compounds enhanced apolipoprotein A-I/ABCA1-dependent generation of HDL in vitro, and increased hepatic ABCA1 and plasma HDL without increasing antioxidant activity in plasma when given to rabbits. Both compounds also decreased vascular lipid deposition in cholesterol-fed rabbits. We therefore conclude that stabilization of ABCA1 against calpain-mediated degradation is a novel and potentially important strategy to increase HDL formation and prevent atherosclerosis. Spiroquinone and diphenoquinone are potential seeds for development of such drugs.

The purpose of this study is to make clear the polishing characteristics of the manganese oxide slurry for wire material of Cu. To examine the polishing characteristics of the manganese oxide slurry, a series of polishing experiments for oxygen free copper have been carried out by using MnO2, Mn2O3 and Al2O3 as abrasive grains and polishing properties have been discussed. As a result, the best result among three kinds of slurries is obtained in polishing with Mn2O3 slurry at the viewpoints of the stock removal and surface roughness. Furthermore polished surfaces have been analyzed using SEM and atomic force microscope. While a large number of mechanical scratches are observed in the polished surface with Al2O3 slurry, the polished surfaces with manganese oxide slurries are covered with fine protrusions. It seems that the fine protrusions result from interaction between mechanical and chemical effects.

We examined the influence of imipramine, a traditional tricyclic antidepressant, on the binding to serotonin (5-HT)2receptors and levels of 5-HT2A–receptor mRNA in the frontal cortex of rats treated with adrenocorticotropic hormone (ACTH). Chronic treatment with ACTH significantly increased the binding of [3H]-ketanserin to 5-HT2receptors and the expression of 5-HT2A–receptor mRNA in the frontal cortex. However, it did not alter the concentration of 5-HT or 5-hydroxyindole acetic acid. The effect of chronic ACTH treatment on 5-HT2receptor and 5-HT2A–receptor mRNA levels was not altered by the chronic administration of imipramine. Also, imipramine did not affect the hyperfunction of 5-HT2Areceptors caused by chronic ACTH treatment. These findings suggest that chronic treatment with ACTH acts to increase 5-HT2A– receptor synthesis through increased gene transcription, without modulating presynaptic serotonergic neurotransmission. Keywords:: 5-HT2Areceptor, adrenocorticotropic hormone, wet-dog shake, receptor binding, receptor mRNA

Atherothrombotic disease is a leading public health problem. Although current antiplatelet agents, such as aspirin and adenosine diphosphate (ADP)-receptor antagonists, reduce the morbidity and mortality associated with atherothrombotic disease, the residual risk for ischemic events remains substantial. The high residual risk despite dual antiplatelet therapy can be attributed to the fact that platelets possess multiple pathways of activation that are not all inhibited by aspirin and ADP-receptor antagonists. Among these, binding of thrombin to the proteinase-activated receptor 1 (PAR1) is the most potent platelet activation pathway. In addition, the PAR1 pathway does not appear to be essential for initiating hemostasis. Inhibition of the PAR1 receptor thus offers a possible new therapeutic approach with a potentially improved benefit-torisk profile for treatment of patients with atherothrombotic disease. Preclinical and clinical studies have confirmed that SCH 530348, a potent, orally active thrombin-receptor antagonist selective for PAR1, does not increase bleeding liability when added to dual antiplatelet therapy. Currently, two large ongoing phase 3 clinical trials are evaluating the efficacy and safety of SCH 530348 in combination with the standard of care in patients with acute coronary syndromes as well as for secondary prevention in patients with previous history of atherothrombotic disease. Keywords:: atherothrombosis, antiplatelet therapy, proteinase-activated receptor, 1 (PAR1), thrombin-receptor antagonist, SCH 530348

Micro RNAs (miRNAs) regulate the expression of target genes posttranscriptionally by pairing incompletely with mRNA in a sequence-specific manner. About 30% of human genes are regulated by miRNAs, and a single miRNA is capable of reducing the production of hundreds of proteins by means of incomplete pairing upon miRNA-mRNA binding. Lately, evidence implicating miRNAs in the development of lung cancers has been emerging. In particular, miR-19a, which is highly expressed in malignant lung cancer cells, is considered the key miRNA for tumorigenesis. However, its direct targets remain underreported. In the present study, we focused on six potential miR-19a target genes selected by miRNA target prediction software. To evaluate these genes as direct miR-19a target genes, we performed luciferase, pull-down, and western blot assays. The luciferase activity of plasmids with each miR-19a-binding site was observed to decrease, while increased luciferase activity was observed in the presence of anti-miR-19a locked nucleic acid (LNA). The pull-down assay showed biotinylated miR-19a to bind to AGO2 protein and to four of six potential target mRNAs. Western blot analysis showed that the expression levels of the four genes changed depending on treatment with miR-19a mimic or anti-miR-19a-LNA. Finally, FOXP1, TP53INP1, TNFAIP3, and TUSC2 were identified as miR-19a targets. To examine the function of these four target genes in lung cancer cells, LK79 (which has high miR-19a expression) and A549 (which has low miR-19a expression) were used. The expression of the four target proteins was higher in A549 than in LK79 cells. The four miR-19a target cDNA expression vectors suppressed cell viability, colony formation, migration, and invasion of A549 and LK79 cells, but LK79 cells transfected with FOXP1 and TP53INP1 cDNAs showed no difference compared to the control cells in the invasion assay.

Synovial sarcoma is a relatively rare high-grade soft tissue sarcoma that often develops in the limbs of young people and induces the lung and the lymph node metastasis resulting in poor prognosis. In patients with synovial sarcoma, specific chromosomal translocation of t(X; 18) (p11.2;q11.2) is observed, and SS18-SSX fusion protein expressed by this translocation is reported to be associated with pathogenesis. However, role of the fusion protein in the pathogenesis of synovial sarcoma has not yet been completely clarified. In this study, we focused on the localization patterns of SS18-SSX fusion protein. We constructed expression plasmids coding for the full length SS18-SSX, the truncated SS18 moiety (tSS18) and the truncated SSX moiety (tSSX) of SS18-SSX, tagged with fluorescent proteins. These plasmids were transfected in synovial sarcoma SYO-1 cells and we observed the expression of these proteins using a fluorescence microscope. The SS18-SSX fusion protein showed a characteristic speckle pattern in the nucleus. However, when SS18-SSX was co-expressed with tSSX, localization of SS18-SSX changed from speckle patterns to the diffused pattern similar to the localization pattern of tSSX and SSX. Furthermore, cell proliferation and colony formation of synovial sarcoma SYO-1 and YaFuSS cells were suppressed by exogenous tSSX expression. Our results suggest that the characteristic speckle localization pattern of SS18-SSX is strongly involved in the tumorigenesis through the SSX moiety of the SS18-SSX fusion protein. These findings could be applied to further understand the pathogenic mechanisms, and towards the development of molecular targeting approach for synovial sarcoma.

The miR-17-92 cluster encodes 7 miRNAs inside a single polycistronic transcript, and is known as a group of oncogenic miRNAs that contribute to tumorigenesis in several cancers. However, their direct targets remain unclear, and it has been suggested that a single miRNA is capable of reducing the production of hundreds of proteins. The majority of reports on the identification of miRNA targets are based on computational approaches or the detection of altered mRNA levels, despite the fact that most miRNAs are thought to regulate their targets primarily by translational inhibition in higher organisms. In this study, we examined the target profiles of miR-19a, miR-20a and miR-92-1 in MCF-7 breast cancer cells by a quantitative proteomic strategy to identify their direct targets. A total of 123 proteins were significantly increased after the endogenous miR-19a, miR-20a and miR-92-1 were knocked down, and were identified as potential targets by two-dimensional electrophoresis and a mass spectrometric analysis. Among the upregulated proteins, four (PPP2R2A, ARHGAP1, IMPDH1 and NPEPL1) were shown to have miR-19a or miR-20a binding sites on their mRNAs. The luciferase activity of the plasmids with each binding site was observed to decrease, and an increased luciferase activity was observed in the presence of the specific anti-miRNA-LNA. A Western blot analysis showed the expression levels of IMPDH1 and NPEPL1 to increase after treatment with anti-miR-19a, while the expression levels of PPP2R2A and ARHGAP1 did not change. The expression levels of IMPDH1 and NPEPL1 did not significantly change by anti-miR-19a-LNA at the mRNA level. These results suggest that the IMPDH1 and NPEPL1 genes are direct targets of miR-19a in breast cancer, while the exogenous expression of these genes is not associated with the growth suppression of MCF-7 cells. Furthermore, our proteomic approaches were shown to be valuable for identifying direct miRNA targets.