Ehime University Hospital

561 Background: The expression of prostate-specific membrane antigen (PSMA) protein increases as prostate cancer advances and metastasizes. Recently, 68Ga-labeled PSMA PET has been used in diagnosing prostate cancer. Our previous studies demonstrated that PSMA-positive vesicles secreted by prostate cancer cells enhance the angiogenic activity of vascular endothelial cells, suggesting a role for PSMA in tumor angiogenesis. In this study, we focus on renal cell carcinoma, examining the relationship between PSMA expression in peritumoral blood vessels and clinical outcomes, particularly tumor recurrence, in surgically excised renal cancer specimens. Methods: Immunohistochemical analysis was performed on renal cell carcinoma samples from 45 patients who underwent nephrectomy or partial nephrectomy at our institution between January 2018 and December 2022. The correlation between PSMA expression levels in tumor-associated vessels and the rate of recurrence was evaluated. In addition, PSMA levels in human umbilical vein endothelial cells (HUVECs) treated with conditioned medium (CM) from renal cancer cell lines Caki1 and ACHN were analyzed by Western blotting (WB) and immunofluorescence (IF). Angiogenic activity was measured using a tube formation assay, and RNA sequencing (RNA-seq) was conducted to examine gene expression changes in endothelial cells influenced by vesicles secreted from renal cancer cells. Spatial gene expression analysis was performed to compare differences in gene expression between tumor vessels near and vessels away from the tumor. Results: Immunohistochemical staining for PSMA and CD31 revealed that only peritumoral vessels exhibited PSMA positivity. PSMA expression was categorized into three levels: weak (18 cases), moderate (11 cases), and strong (16 cases), with these levels corresponding to different recurrence rates. When the 10,000 g pellet fraction of CM from renal cancer cell lines was applied to HUVECs, they became PSMA-positive, leading to enhanced tube formation. RNA-seq analysis further demonstrated that HUVECs exposed to CM from renal cancer cells showed upregulation of pathways associated with angiogenesis. Conclusions: Vesicles secreted by renal cancer cells promote PSMA expression in vascular endothelial cells and boost angiogenic activity. Future research will aim to elucidate the mechanism by which PSMA becomes expressed in normal vascular endothelial cells, with the potential to pave the way for new anti-angiogenic therapies.

AbstractAimPortal hypertension resulting from liver cirrhosis (LC) can lead to pancreatic congestion and impaired insulin secretion. Therefore, this prospective study aimed to assess the association between pancreatic congestion and exocrine pancreatic function in patients with LC.MethodsIn our clinical study, pancreatic congestion and exocrine pancreatic function were evaluated using shear wave dispersion (SWD) and fecal elastase‐1 (FE‐1). Additionally, pancreatic acinar cells, venous walls, and fibrosis were assessed in an autopsy study.ResultsThe FE‐1 levels were lower in the LC group (n = 41) than in the control group (n = 41) (312 ± 89 vs. 442 ± 100 μg/g, p < 0.001). The LC group included six patients (14.6%) with exocrine pancreatic insufficiency, whereas there were none in the control group. Pancreatic SWD values were significantly higher in the LC group than in the control group (14.8 ± 2.3 vs. 10.0 ± 1.28 [m/s]/kHz, p < 0.001). Fecal elastase‐1 was significantly negatively correlated with pancreatic SWD (R = −0.55, p < 0.001). As for the autopsy study, the percentage of the trypsin‐positive area was significantly lower in the LC group (n = 11) than in the control group (n = 10) (38.1 ± 10.1% vs. 26.5 ± 3.0%, p = 0.0055). The percentage of trypsin‐positive area was significantly negatively correlated with the wall thickness of the pancreatic vein (R = −0.76, p < 0.001).ConclusionsExocrine pancreatic function was reduced and significantly correlated with pancreatic congestion in patients with LC. Portal hypertension may affect the exocrine pancreatic function.

AbstractObjectivesEpidemiologic evidence on the association between intake of milk and dairy products and dental caries is limited, particularly in Asia. This cross‐sectional study examined the association between the consumption of milk and dairy products and dental caries among Japanese children aged 3 years.MethodsThe study subjects were 6221 children. Parents or guardians completed a questionnaire, including a self‐administered food frequency questionnaire for children. Dentists assessed dental caries, and these data were recorded in each child's maternal and child health handbook. Parents or guardians transcribed these data from the handbook to our questionnaire. Children were classified as having dental caries if they had one or more decayed or filled primary teeth. Associations with dental caries were assessed using logistic regression analysis with adjustments in demographics, dietary and lifestyle factors, and parental socioeconomic status.ResultsThe prevalence of dental caries was 14.6%. Intakes of milk, cheese, and yogurt were associated with 21%, 26%, and 35% decreases, respectively, in the odds of the prevalence of dental caries (p = 0.02, 0.001, and 0.002, respectively), whereas the intake of other dairy products, such as probiotic milk, ice cream, or custard pudding, was associated with a 2.3‐fold increase in the odds of the prevalence of dental caries (p < 0.0001). There was no association between intake of total dairy products and dental caries.ConclusionsConsumption of milk, cheese, or yogurt had a beneficial effect on childhood dental caries, even in Japan where people consume relatively less milk and dairy products.

Abstract Purpose To evaluate the effects of four-dimensional noise reduction filtering using a similarity algorithm (4D-SF) on the image quality and tumor visibility of low-dose dynamic computed tomography (CT) in evaluating breast cancer. Materials and methods Thirty-four patients with 38 lesions who underwent low-dose dynamic breast CT and were pathologically diagnosed with breast cancer were enrolled. Dynamic CT images were reconstructed using iterative reconstruction alone or in combination with 4D-SF. We selected the peak enhancement phase image of breast cancer for each patient for quantitative and qualitative evaluations of image quality and measurement of the maximum diameter of breast cancer. The signal-to-noise and contrast-to-noise ratios were calculated for quantitative evaluation. The maximum diameters of the breast cancer were measured from the images obtained with and without 4D-SF (4D-SF ±) (size-4D-SF + and size-4D-SF-) and for the pathological specimen (size-PS) and compared. Results The median and interquartile ranges of the signal-to-noise ratio [4D-SF-: 3.03 (2.54–4.17) vs 4D-SF + : 5.52 (4.75–6.66)] and contrast-to-noise ratio [4D-SF-: 2.88 (2.00–3.60) vs 4D-SF + : 7.84 (4.65–10.35)] were significantly higher for 4D-SF + than for 4D-SF- (p < 0.001). The overall image quality (Observer 1, p < 0.001; Observer 2, p < 0.001) and tumor margin sharpness scores (Observer 1, p = 0.003; Observer 2, p < 0.001) were significantly higher for 4D-SF + than for 4D-SF-. The tumor contrast scores for 4D-SF + and 4D-SF- were not significantly different (Observers 1, 2; p = 0.083). Size-4D-SF- was significantly smaller than size-PS (p < 0.001); size-4D-SF + was also smaller than size-PS, but the difference was not significant (p = 0.088). The Spearman’s rank correlation coefficient was 0.65 for size-PS and size-4D-SF- and 0.77 for size-PS and size-4D-SF + . Conclusion The 4D-SF can improve the image quality and tumor visibility of low-dose dynamic CT in evaluating breast cancer extent due to noise reduction.

Prostate-specific membrane antigen (PSMA) protein expression is induced during prostate cancer progression and metastasis. Recently, we reported that PSMA-positive vesicles released by prostate cancer cell lines enhanced vascular endothelial cell angiogenesis and that PSMA may be involved in tumor angiogenesis. Similarly, it is known that PSMA is upregulated in peritumoral vessels in renal cell carcinoma (RCC). In this study, we investigated the significance and molecular function of PSMA in RCC. PSMA immunohistochemical staining confirmed PSMA presence only in perinephric tumor vessels, and PSMA intensity was strongly correlated with recurrence rate and venous invasion. Spatial gene expression analysis revealed that FOLH1 expression, which codes PSMA, was upregulated in tumor blood vessels around renal cancer, and that angiogenesis-related pathways were enhanced. The 10,000 g pellet fraction of the renal cancer cell lines Caki1- and ACHN-conditioned medium (CM) induced PSMA positivity in human umbilical vein endothelial cells (HUVECs) and enhanced tube formation. Mass spectrometry indicated that the 10,000 g pellet fraction contained various kinds of growth factors, like GDF15 and MYDGF. RNA sequencing showed that supplementing HUVECs with RCC cell CM-enhanced angiogenesis-related signaling pathways. Conclusively, microvesicle components secreted by RCC cells transform vascular endothelial cells into PSMA-positive cells, enhancing angiogenesis.

Allergic diseases and ulcerative colitis (UC) share pathophysiological similarities. This study aimed to investigate the unclear association between allergic diseases and mucosal healing (MH), an important factor in the prognosis of UC. We studied 289 Japanese patients with UC. Information on allergic diseases (bronchial asthma, atopic dermatitis, pollen allergy, food allergy, and drug allergy), as diagnosed by physicians, was collected through self-reported questionnaires. The definition of MH was Mayo Endoscopic Score 0. The association between each allergic disease and its multimorbidity with MH was evaluated using multivariate logistic regression analyses. Pollen allergy was the most common allergic condition (36.3%). Pollen allergy and food allergy were independently associated with MH (pollen allergy adjusted OR: 1.82 [95% CI: 1.01–3.26]; food allergy adjusted OR: 3.47 [95% CI: 1.26–9.68]). The rates of MH for 0 and 3 or more allergic diseases were 24.6% and 4.2%, respectively. After adjustment for confounders, 3 or more allergic diseases were independently associated with MH (adjusted OR: 8.13 [95% CI: 2.17–34.04], p for trend = 0.020). This study demonstrates a significant positive association between specific allergic diseases (pollen and food allergies) and MH in UC patients, with a stronger association in cases of allergic multimorbidity.

AbstractBone is a unique organ crucial for locomotion, mineral metabolism, and hematopoiesis. It maintains homeostasis through a balance between bone formation by osteoblasts and bone resorption by osteoclasts, which is regulated by the basic multicellular unit (BMU). Abnormal bone metabolism arises from an imbalance in the BMU. Osteoclasts, derived from the monocyte‐macrophage lineage, are regulated by the RANKL‐RANK‐OPG system, which is a key factor in osteoclast differentiation. RANKL activates osteoclasts through its receptor RANK, while OPG acts as a decoy receptor that inhibits RANKL. In trabecular bone, high turnover involves rapid bone formation and resorption, influenced by conditions such as malignancy and inflammatory cytokines that increase RANKL expression. Cortical bone remodeling, regulated by aged osteocytes expressing RANKL, is less understood, despite ongoing research into how Rett syndrome, characterized by MeCP2 abnormalities, affects RANKL expression. Balancing trabecular and cortical bone involves mechanisms that preserve cortical bone, despite overall bone mass reduction due to aging or oxidative stress. Research into genes like sFRP4, which modulates bone mass, highlights the complex regulation by BMUs. The roles of the RANKL‐RANK‐OPG system extend beyond bone, affecting processes such as aortic valve formation and temperature regulation, which highlight the interconnected nature of biological research.

Objectives: While the effectiveness of metabolic/bariatric surgery has been confirmed, understanding the factors associated with weight loss is paramount for providing guidance in postoperative treatment strategies. Here, we aimed to examine the factors associated with long-term maintenance of weight loss after laparoscopic sleeve gastrectomy (LSG). Methods: This prospective observational cohort included patients who underwent LSG at a single academic health center between January 2017 and June 2022. We examined their body composition using InBody 720 or 770 and analyzed the factors associated with the percentage of total weight loss (%TWL) for 24 months. Results: The median body mass index (BMI) was 38.8 (interquartile range [IQR]: 35.6–46.7) preoperatively, 32.7 kg/m2 (IQR: 28.2–38.7) at 12 months postoperatively, and 33.9 kg/m2 (IQR: 29.1–40.1) at 24 months postoperatively. The lowest BMI was observed at 12 months (p < 0.001 vs. preoperative), followed by a significant increase at 24 months (p = 0.003). However, BMI remained significantly lower at 24 months than preoperatively (p < 0.001). The skeletal muscle mass to fat mass ratio (SMM/FM) was 0.59 (IQR: 0.50–0.71) preoperatively, 0.79 (IQR: 0.58–1.26) at 12 months, and 0.70 (IQR: 0.54–1.05) at 24 months, peaking at 12 months (p < 0.001 vs. preoperative) and decreasing significantly by 24 months (p < 0.001). Nevertheless, the SMM/FM ratio at 24 months remained higher than preoperative values (p < 0.001). Median body weight and %TWL were 86.0 kg and 15.6%, respectively, at 24 months after LSG. The SMM/FM ratio at 12 months was positively correlated with %TWL at 24 months after adjusting for age and sex. Conclusions: The effects of LSG persisted for up to 24 months postoperatively. The SMM/FM ratio 12 months after LSG was associated with the rate of weight loss at 24 months.

Abstract Background A simple suspension method, where solid formulations are disintegrated and suspended by being soaked in warm water followed by tube administration, is widely used, especially for elderly patients with dysphagia in Japanese clinical settings. However, there is insufficient information on drug stability in the simple co-suspension of multiple formulations especially including acidic or alkaline ones. The influence of occasional prolonged soakage on drug stability is also of concern. In this study, the chemical stability of typical β-lactam antibiotics, amoxicillin, and cefcapene pivoxil hydrochloride, was investigated in simple co-suspensions with magnesium oxide (MgO), which is frequently used as an alkaline laxative for the elderly. Methods Amoxicillin (capsule) or cefcapene pivoxil hydrochloride (tablet) was placed with or without MgO (tablet) in a centrifuge tube containing warm water (55 °C). The tube was allowed to stand for 10 min or 5 h at room temperature and simple suspensions were prepared. The suspensions were then treated with large amounts of solvents and neutralized using a weakly acidic cation exchange resin. The resulting solutions were analyzed by high-performance liquid chromatography. The degradation products were identified by mass spectrometry and nuclear magnetic resonance spectroscopy. Results Amoxicillin was found to be partially degraded to amoxicilloic acid and amoxicillin diketopiperazine by the co-suspension with MgO. The degree of degradation increased with the prolonged soaking. The recovery rates of cefcapene pivoxil decreased due to the poor solubility in the co-suspensions with MgO and no degradation product of the drug was observed. Conclusions Amoxicillin and MgO should be independently suspended because of the chemical instability of amoxicillin. This study has also indicated there is a degradation risk after prolonged soaking. It should be noted that the poor water solubility of cefcapene pivoxil under alkaline conditions may affect the absorption process as well as tube passability.

RNA-dependent protein kinase (PKR) may have a positive regulatory role in controlling tumor growth and progression in hepatocellular carcinoma (HCC). However, the downstream substrates and the molecular mechanism of PKR in the growth and progression of HCC have not been clarified. In this study, mass spectrometry analysis was performed with immunoprecipitated samples, and 4.1R was identified as a protein that binds to PKR. In transfected COS7 cells, an immunoprecipitation experiment showed that 4.1R binds to wild-type PKR, but not to a kinase-deficient mutant PKR, suggesting that PKR binds to 4.1R in a kinase activity-dependent manner. In HCC cell lines, HuH7 and HepG2, the expression level of 4.1R protein was shown to be regulated by protein expression and activation of PKR. Interestingly, high expression of 4.1R, as well as PKR, is associated with a worse prognosis in HCC. PKR increased HCC cell growth in both anchorage-dependent and anchorage-independent manners, whereas 4.1R was involved in HCC cell growth only in an anchorage-independent manner, not in an anchorage-dependent manner. The rescue experiment indicated that increased anchorage-independent growth of HCC cells by PKR might be caused by 4.1R. In conclusion, PKR associates with 4.1R and promotes anchorage-independent growth of HCC. The PKR-4.1R axis might be a new therapeutic target in HCC.

Introduction: Avatrombopag (AVA) is an orally administered thrombopoietin receptor (TPO-RA) agonist widely approved for the treatment of adults with chronic immune thrombocytopenia (ITP). This study aims to assess AVA in Japanese patients with chronic ITP. Methods: This Phase 3, multicenter, open-label study (a 26-week core phase [completed] followed by an extension phase [ongoing until market authorization in Japan]) evaluates the efficacy and safety of AVA (initial dose 20 mg/day) in Japanese adults (aged ≥18 years) with chronic ITP (≥12 months duration), insufficient response to prior treatment (investigator assessed), and an average of two platelet counts (PCs) <30×109/L. The core phase included a 1-day baseline assessment, a 6-week dosage titration phase, a 12-week concomitant medication reduction phase, and 8 weeks of maintenance treatment. The primary endpoint was the cumulative number of weeks in which the PC was ≥50×109/L during 26 weeks of treatment in the absence of rescue therapy. The key secondary endpoint was the platelet response rate at Day 8 (percentage of patients with a PC ≥50×109/L). Additional efficacy endpoints included durable platelet response (six of eight weekly PC ≥50×109/L during the last 8 weeks of treatment in the absence of rescue therapy), continuous platelet response (the number of consecutive weeks of platelet response ≥50×109/L in the absence of rescue therapy), International Working Group (IWG) complete platelet response (PC ≥100×109/L and absence of bleeding or rescue therapy at each analysis visit), IWG platelet response (PC ≥30×109/L and at least a two-fold increase in baseline platelet count and absence of bleeding or rescue therapy at each analysis visit), and concomitant ITP medication reduction/discontinuation. The primary, key secondary, and additional efficacy endpoints, and the maximum grade of treatment-emergent adverse events [TEAEs] and bleeding events in the core phase [using the World Health Organization (WHO) Bleeding Scale]) are reported. Results: In total, 19 patients (mean age 56.0 years; 78.9% female) were enrolled at 19 sites in Japan; 15 patients (78.9%) completed the core phase, and four patients (21.1%) discontinued the study (due to: adverse events [one patient], lack of efficacy at the highest dose [one patient], and use of prohibited concomitant medication [two patients]). Eight patients (42.1%) had a baseline PC ≤15×109/L, and nine patients (47.4%) used concomitant ITP medication at baseline. Three patients (15.8%) had at least one previous significant bleeding event prior to the core phase. The mean cumulative number of weeks of platelet response was 13.47 weeks (95% confidence interval [CI]: 9.13, 17.80). At Day 8, 12 patients (63.2%; 95% CI: 38.4, 83.7) had a platelet response (PC ≥50×109/L), and eight patients (42.1%; 95% CI: 20.3, 66.5) had a durable platelet response during the last 8 weeks of the core phase. The mean maximum duration of continuous platelet response was 7.63 weeks (standard deviation: 6.907). IWG complete platelet response and IWG platelet response were achieved by 5 of 19 (26.3%) and 9 of 19 (47.4%) patients by Day 8, and by 4 of 15 (26.7%) and 9 of 15 (60.0%) patients, respectively, at Week 26. Of the nine patients who used concomitant ITP medications at baseline, five (55.6%) reduced their use, and one patient discontinued use. Most TEAEs were Grade 1 or 2 (15 patients, 79.0%). A Grade 3 TEAE occurred in two patients (diffuse large B-cell lymphoma and heavy menstrual bleeding) and a Grade 4 TEAE (autoimmune hepatitis) occurred in one patient. No deaths or thromboembolic events were reported. Most bleeding events were mild (WHO Grade 1); no patients had a WHO Grade 3 or 4 bleeding event. Conclusion: AVA effectively provided a rapid and durable platelet response and induced complete platelet response by IWG criteria in Japanese patients with chronic ITP. Furthermore, AVA was well tolerated in these patients. These data add to the existing body of evidence supporting the efficacy and safety of AVA in different populations.