[Development of Analytical Method and Surveillance ofGibberellic Acid in Banana, Cherry, and Kiwi Fruit].

Gibberellic acid (GA3) is commonly used as a plant growth regulator in many food crops owing to its essential signaling functions during plant growth and development. In Japan, a threshold for administrative action for GA3 content of 0.3 mg/kg applies in produce in which maximum residue limits have not been established. Although the threshold is based on previous studies, the GA3 concentrations in individual foods are still unknown. Thus, we surveyed the concentrations of GA3 in banana, cherry, and kiwi fruit on the Japanese market. We developed and validated a method for the analysis of GA3 using solid-phase extraction and LC-MS/MS in accordance with accepted criteria of trueness, repeatability, and selectivity. The limits of detection and of quantification were determined as 0.005 and 0.05 mg/kg, respectively, in all fruits. Concentrations of GA3 did not exceed 0.3 mg/kg regardless of ripeness, suggesting the reasonability of the current regulation of GA3 in banana, cherry, and kiwi fruit. These findings can support prompt administrative action on these fruits, contributing to the regulation of GA3 in Japan.

[Inter-laboratory Study on the Modified Methods for Analyzing Bisphenol A Content for Migration Tests from Polycarbonate Food Apparatuses, Containers, and Packaging].

An inter-laboratory study involving 24 laboratories was conducted to validate the modified analytical method for the migration solution of heptane for the determination of bisphenol A migrating from polycarbonate food processing materials. In this study, two concentrations of samples were blindly coded. Each laboratory determined the analyte (bisphenol A, phenol and p-tert-butylphenol) concentration in each sample according to the established protocol. The obtained values were analyzed statistically using internationally accepted guidelines. Horwitz ratios were calculated based on the reproducibility relative standard deviation (RSDR), which was estimated from the inter-laboratory study, and predicted RSDR, which was calculated using the Horwitz/Thompson equation. Horwitz ratios of the two samples ranged from 0.15 to 0.37 for the three compounds, meeting the performance criteria of less than 2 set by the Codex Alimentarius for analytical method approval. These results showed that this modified analytical method shows good performance as an analytical method for the migration solution of heptane.

Student athlete well-being framework: an empirical examination of elite college student athletes.

The current study proposes a multidimensional student athlete well-being framework (SAWBF). The authors used 12 items to capture SAWBF comprised of four well-being dimensions (i.e., physical, hedonic, psychological, and social well-being). To empirically assess the reliability and validity of the framework, data from elite collegiate student athletes in Japan (N = 546) were procured. The results indicated sufficient convergent and discriminant validities of SAWBF. The authors also assessed predictive validity correlations of the framework by focusing on the oft-supported well-being outcome-organizational citizenship behavior, which were also found to be associated with SAWBF. The findings indicated the usefulness of SAWBF; and coaches and staff members can utilize the framework to multi-dimensionally understand well-being status of their student athletes, potentially boosting adaptive behaviors.

[Modified Methods for Analyzing Bisphenol A Content for Migration Tests from Polycarbonate Food Apparatuses, Containers, and Packaging].

A validation study was performed on the modified analytical method for the migration solution of heptane, 20% ethanol and 4% acetic acid for the determination of bisphenol A migrating from polycarbonate food apparatuses, containers, and packaging. The analytes for the method were bisphenol A, phenol and p-tert-butylphenol. The repeatability, within-laboratory reproducibility and trueness of the method was estimated in the range of 0.2-1.8%, 0.4-2.6% and 95-102% respectively. These results showed that the method is useful as an analytical method for the migration solution of heptane, 20% ethanol and 4% acetic acid. Furthermore, the applicability of the determination methods with a fluorescence detector was verified. As a result of the validation study, the repeatability, within-laboratory reproducibility and trueness of the method was estimated in the range of 0.1-2.9%, 0.2-3.1% and 94-101% respectively. It was confirmed that the measurement with a fluorescence detector is also available.

Preoperative C-reactive protein-to-albumin ratio as a prognostic factor in biliary tract cancer: A systematic review and meta-analysis.

BACKGROUND: The preoperative C-reactive protein-to-albumin ratio is a novel inflammation-based prognostic marker in various cancers. However, its prognostic role in biliary tract cancer is unknown. We conducted a systematic review and meta-analysis to evaluate the prognostic value of preoperative C-reactive protein-to-albumin ratio in biliary tract cancer. METHODS: A systematic search of the literature for studies evaluating the prognostic value of C-reactive protein-to-albumin ratio in patients undergoing surgery for biliary tract cancer was conducted, and a random effects meta-analysis of overall survival and recurrence-free survival was performed. RESULTS: Nine studies with 1292 participants were included. The preoperative C-reactive protein-to-albumin ratio negatively correlated with overall survival (hazard ratio, 2.44 [95% confidence interval: 1.98-2.90]; P < .001) and recurrence-free survival (hazard ratio, 2.73 [95% confidence interval: 2.01-3.70]; P < .001). Subgroup analysis showed that an elevated preoperative C-reactive protein-to-albumin ratio predicted poor overall survival, regardless of the cutoff value, sample size, histological type, and treatment. CONCLUSION: An elevated preoperative C-reactive protein-to-albumin ratio is significantly associated with poor prognosis in patients undergoing surgery for biliary tract cancer. The C-reactive protein-to-albumin ratio may be an independent prognostic biomarker for overall survival and recurrence-free survival in patients undergoing surgery for biliary tract cancer.

[Estimation of the Distribution of Dietary Exposure to Lead Using Two-Dimensional Monte Carlo Simulation: An Attempt to Apply to Total Diet Samples].

The average dietary exposure to lead (Pb) in male and female Japanese individuals >1 year of age was estimated using 280 total diet samples representing 14 food groups from 10 areas over a two-year period. A probabilistic exposure estimation was performed using a two-dimensional Monte Carlo simulation (2D-MCS) with a Bayesian estimation that consided the uncertainty of the estimation process. The Bayesian estimation was performed using the likelihood function with cumulative distribution function between the lower and upper boundary values for no-detected values. The median dietary exposure to Pb was estimated as 5.82 µg/person/day. The 90% interval was 2.51-16.9 µg/person/day. Comparison with previously reported Pb exposure values indicates that the estimation of Pb exposure distribution using total diet samples is reasonable. The contribution to Pb exposure was highest in the order of food group 8 (light-colored vegetables, mushrooms, and seaweeds: 20.0±16.1%)>food group 1 (rice and rice products: 12.3±19.0%)>food group 10 (fish and shellfish: 10.5±13.9%). Owing to the high uncertainties of contribution ratios, it was not possible to identify dominant food groups contributing to Pb exposure. However, it was evident that the uncertainty of the estimation of Pb exposure was influenced by the uncertainty of Pb concentration than the uncertainty of food consumption rate. In particular, the effect of uncertainty from the Pb concentration of the food group 1 was 68.2%. When the margin of exposures were calculated, the estimated probabilities that a value would be <1 were 14.5% for developmental neurotoxicity to children (1-6 years old), 0.13% for blood pressure and 0.93% for kidney disease in Japanese individuals ≥1 year of age. The findings suggest that the health risk due to dietary Pb exposure is small but not negligible.

Lymphocyte-to-C-Reactive Protein Ratio Predicts Prognosis in Patients With Colorectal Liver Metastases Post-hepatic Resection: A Retrospective Study.

BACKGROUND/AIM: Preoperative systemic inflammation has been reported to predict survival in patients with various cancer types. In patients with colorectal liver metastasis (CRLM), the prognosis is poor despite therapeutic advances in the field. Here, we aimed to evaluate the prognostic role of the lymphocyte-to-C-reactive protein (CRP) ratio (LCR) in patients with CRLM after hepatic resection. PATIENTS AND METHODS: This retrospective study included 104 patients who underwent hepatic resection for CRLM between October 2010 and 2021 at the National Hospital Organization Fukuyama Medical Center, Hiroshima, Japan. The association between clinicopathological variables, including various inflammatory biomarkers [LCR, platelet-to-lymphocyte ratio (PLR), neutrophil-to-lymphocyte ratio (NLR), CRP-to-albumin ratio (CAR), and prognostic nutritional index (PNI)], and overall survival of the patients was investigated using univariate and multivariate analyses. RESULTS: The optimal cut-off values for each biomarker by receiver-operating characteristic analysis were as follows: LCR: 12,720; PLR: 150; NLR: 4; CAR: 0.023; and PNI: 44.8. The 1-, 3-, and 5-year overall survival rates were 97.0%, 71.3%, and 56.8%, respectively. On univariate analysis, LCR<12, 720, PLR<0.14, body mass index <24 kg/m2, carbohydrate antigen 19-9 ≥37 U/ml, multiple tumours, and largest hepatic tumour ≥5 cm were significant factors predictive of poorer survival. The multivariate analysis revealed that LCR<12, 720 (hazard ratio=2.156, 95% confidence interval=1.060-4.509, p=0.034) and multiple tumours (HR=2.336, 95% CI=1.125-4.925, p=0.023) were independent predictors of poor overall survival. CONCLUSION: LCR may be an independent prognostic predictor in patients after hepatic resection for CRLM. Therefore, the assessment of LCR as a biomarker may help in treatment planning.

Preoperative lymphocyte-to-C-reactive protein ratio predicts hepatocellular carcinoma recurrence after surgery.

Purpose: This study was performed to determine the prognostic value of lymphocyte-to-CRP ratio after curative resection for hepatocellular carcinoma. Methods: Between July 2010 and October 2021, 173 consecutive patients (144 male, 29 female) who underwent surgical resection for pathologically confirmed hepatocellular carcinoma were included in this retrospective study. Cox regression analysis was used to evaluate the relationship between clinicopathological characteristics and recurrence-free survival (RFS) and overall survival (OS). A P-value of <0.05 was considered statistically significant. Results: The patients (mean age, 71 years) were stratified into high (≥9,500, n = 108) and low (<9,500, n = 65) lymphocyte-to-CRP ratio groups. The low lymphocyte-to-CRP ratio group had significantly worse RFS and OS. Low lymphocyte-to-CRP ratio (hazard ratio [HR], 1.865; 95% confidence interval [CI], 1.176-2.960; P = 0.008), multiple tumors (HR, 3.333; 95% CI, 2.042-5.343; P < 0.001), and microvascular invasion (HR, 1.934; 95% CI, 1.178-3.184; P = 0.009) were independently associated with RFS, whereas low albumin-to-globulin ratio (HR, 2.270; 95% CI, 1.074-4.868; P = 0.032), α-FP of ≥25 ng/mL (HR, 2.187; 95% CI, 1.115-4.259; P = 0.023), and poor tumor differentiation (HR, 2.781; 95% CI, 1.041-6.692; P = 0.042) were independently associated with OS. Lymphocyte-to-CRP ratio had a higher area under the curve (0.635) than other inflammation-based markers (0.51-0.63). Conclusion: Lymphocyte-to-CRP ratio is superior to other inflammation-based markers as a predictor of RFS in patients with surgically resected hepatocellular carcinoma.

Presence of nano-sized mercury-containing particles in seafoods, and an estimate of dietary exposure.

The toxicity of nano-sized particles of mercury (NP-Hg), which are thought to be generated during the detoxification of methyl mercury (MeHg), may differ from that of MeHg, elemental Hg (Hg0), and inorganic Hg (I-Hg). From a human health perspective, it is important to evaluate the presence of NP-Hg in seafoods. We investigated the in vivo formation of NP-Hg in fish and shellfish, which are the main sources of Hg exposure in humans. NP-Hg was measured in 90 fish samples with single-particle inductively coupled plasma mass spectrometry (spICP-MS) after enzyme degradation with pancreatin and lipase. In addition to NP-Hg, total Hg (T-Hg), MeHg, and selenium (Se) concentrations were evaluated. Transient Hg signals were detected as nanoparticles from almost all samples by using spICP-MS. Higher particle number concentrations (CPN) were observed in the tuna-swordfish group than in the shellfish group (17.7 × 107 vs. 1.2 × 106 particles/g, respectively). Although the CPN and maximum particle mass increased significantly with increasing T-Hg concentration, the increase in CPN was greater than those in maximum particle mass. Assuming that the NP-Hg detected was HgSe (tiemannite) and spherical based on previous reports, the maximum particle diameter was estimated to be 89 nm. The mean dietary exposures to NP-Hg, T-Hg, and MeHg were estimated to be 0.067, 5.75, and 5.32 µg/person per day, respectively. Generation of NP-Hg was inferred to be widespread in marine animals, with a preferential increase in the number of particles rather than an increase in particle size. The mean dietary exposure to NP-Hg in Japanese people was estimated to be 1.2 ng/kg body weight (BW) per day. Compared to PTWI of 4 µg/kg BW per week (0.57 µg/kg BW per day) derived by JECFA (2011), the health risk from redissolved I-Hg from NP-Hg is small.

[Pharmacological characteristics and clinical study results of Pemigatinib (Pemazyre® Tablets), a selective fibroblast growth factor receptor (FGFR) inhibitor].

Pemigatinib (Pemazyre® Tablets 4.5 mg) is a novel fibroblast growth factor receptor (FGFR) inhibitor, created by Incyte Corporation. The product was approved in March 2021 and was launched in June 2021 for the treatment of patients with locally advanced or metastatic biliary tract cancer (BTC) with a fibroblast growth factor receptor 2 (FGFR2) fusion or rearrangement that has progressed after at least one prior line of systemic therapy. Pemigatinib was shown to selectively inhibit kinase activity of FGFR1～3 (IC50; 0.39～1.2 nM). In cultured cells, pemigatinib inhibited the phosphorylation of FGFR1 and its downstream signals, ERK1/2 and STAT5 in a concentration-dependent manner. Pemigatinib also potently inhibited the growth of various types of cell lines with FGFR 1～3 gene alteration. Pemigatinib was shown to induce concentration-dependent tumor regression in a tumor xenograft model mice in which tumor tissue sections from patients with cholangiocarcinoma (CCA) harboring FGFR2 gene fusions were transplanted. Pemigatinib was well tolerated in Japanese and overseas Phase1 studies (INCB 54828-101 and 202). In the global phase2 study (INCB 54828-202) conducted in CCA patients with FGFR2 gene fusions or rearrangements, significant improvement in the overall response rate was observed. Although several adverse reactions were observed which was based on the mechanism of action of pemigatinib, the safety profile and management of the adverse reactions were favorable. Pemigatinib is expected to contribute to second-line drug treatment after failure of standard therapies in biliary tract cancer.

Quantification of progesterone in beef with marbling using liquid chromatography-tandem mass spectrometry with stable isotope-labelled standards.

Progesterone (P4) is contained naturally in animal tissue, and it is also used as a veterinary drug in cattle for treatment purposes. To assess the risk from P4 residues in beef derived from treated cattle, it is essential to quantify the P4 contained naturally in cattle tissue (endogenous P4). Therefore, we performed a method validation for the quantification of endogenous P4 (method quantification limit = 0.06 ng g-1) by using isotope-labelled P4s, and investigated the P4 contents in Japanese beef (n= 112; 0.07 to 121 ng g-1). The P4 contents in cattle muscle ranged from 0.07 to 54.3 ng g-1 in males, and from 0.27 to 121 ng g-1 in females. Our investigation also indicated that the developed method using both 13C- and deuterium-labelled P4 standards could be used to certify the recovery of P4 from cattle muscle containing various amounts of intramuscular fat, and enabled the determination of the P4 content in all Japanese beef samples that exceeded the method quantification limit.

Switching to nilotinib in patients with chronic myeloid leukemia in chronic phase with molecular suboptimal response to frontline imatinib: SENSOR final results and BIM polymorphism substudy.

Optimal management of patients with chronic myeloid leukemia in chronic phase with suboptimal molecular response (MR) to frontline imatinib is undefined. We report final results from SENSOR, which evaluated efficacy/safety of nilotinib in this setting. A substudy assessed whether BIM polymorphisms impacted response to nilotinib. In this single-arm, multicenter study, Japanese patients with suboptimal MR per European LeukemiaNet 2009 criteria (complete cytogenetic response, but not major MR [MMR]) after ≥18 months of frontline imatinib received nilotinib 400mg twice daily for 24 months. MR, BCR-ABL1 mutations/variants, and BIM polymorphisms were evaluated in a central laboratory. Primary endpoint was the MMR rate at 12 months (null hypothesis of 40%). Of 45 patients (median exposure, 22.08 months), 39 completed the study and six discontinued. At 12 and 24 months, 51.1% (95% CI, 35.8%-66.3%) and 66.7% (95% CI, 51.0%-80.0%) achieved MMR, respectively. Cumulative MMR incidence by 24 months was 75.6%. Of 40 patients analyzed, 10 of 12 (83.3%) with and 17 of 28 (60.7%) without BIM polymorphisms achieved MMR at 24 months. The safety profile was manageable with dose reductions and interruptions. Nilotinib provided clinical benefit for patients with suboptimal response to imatinib, and BIM polymorphisms did not influence MMR achievement. ClinicalTrials.gov: NCT01043874.

Alginic acid-based macromolecular chemiluminescent probe for universal protein assay on a solid-phase membrane.

A novel chemiluminescent (CL) technique for the rapid determination of proteins on a membrane is described. The method utilizes an interaction between luminol-labeled alginic acid macromolecule and proteins. The synthesis of the macromolecular probe consists of the oxidation of alginic acid with NaIO(4), the introduction of luminol through imine formation as a CL tag, and the reduction of the conjugate with NaBH(4) to obtain the stable probe. The analytical protocol consists of adsorbing proteins on a poly(vinylidene difluoride) (PVDF) membrane, incubating the membrane for 30 min with the probe solution in the presence of boric acid and a surfactant, two short washing steps in order to remove an excess of the probe, and detection of CL intensity with hemin, tetra-n-propylammonium hydroxide and H(2)O(2). This proposed CL assay for proteins can be finished within 1 h, and indicates the detection limit of 15-250 ng of proteins on the membrane. The CL signals in the calibration curves for some proteins such as albumin show proportional intensities against the amounts of the proteins less than ca. 125 ng, though there is a logarithmic relationship between the CL signals and the protein amounts larger than ca. 125 ng. However, some other proteins indicate the proportional CL intensities against the increasing amounts in wider range up to 500 ng of the proteins. The synthesised alginic acid-based probe indicates specific selectivity towards proteins, and should be used as a CL probe for the universal detection of various proteins on a solid-phase membrane even in the presence of DNA and RNA.

A safety, pharmacokinetic and pharmacodynamic investigation of deferasirox (Exjade, ICL670) in patients with transfusion-dependent anemias and iron-overload: a Phase I study in Japan.

The pharmacokinetics (PK) and pharmacodynamics (PD) of the once-daily, oral ironchelating agent, deferasirox (Exjade, ICL670), have been evaluated further in a Phase I, openlabel, multicenter, dose-escalation study in Japanese patients with myelodysplastic syndromes, aplastic anemia, and other anemias. Deferasirox was initially administered as a single dose of 5 (n = 6), 10 (n = 7), 20 (n = 6) or 30 (n = 7) mg/(kg day) and then after 7 days seven daily doses were administered. Linear PK (C (max) and AUC) were observed at all doses after a single dose and at steady state, and dose-dependent iron excretion was observed. Pharmacokinetic/pharmacodynamic parameters were similar to those reported in a Caucasian beta-thalassemia cohort. Following the single- and multiple-dose phases, 21 of 26 patients progressed to a 3-year extension phase of the study, where dose reductions and increases [5-30 mg/(kg day)] were allowed following safety and efficacy assessments. In the interim, 1-year data show that deferasirox was well tolerated, with generally infrequent and mild adverse events. Reductions in serum ferritin levels were observed and a negative iron balance achieved at doses of 20-30 mg/(kg day). These data suggest that deferasirox has a stable and predictable PK/PD profile, irrespective of underlying disease or race, and a predictable and manageable safety profile suitable for chronic administration.

[Preclinical and clinical profile of imatinib mesilate, a potent protein-tyrosine kinase inhibitor for CML therapy].

Imatinib mesilate (Glivec) is a protein-tyrosine kinase inhibitor that potently inhibits the Bcr-Abl tyrosine kinase as well as the receptors for platelet-derived growth factor (PDGF) and stem cell factor (SCF), c-Kit, at in vitro and cellular kinase assay levels. Since Bcr-Abl tyrosine kinase plays a key role in chronic myelogenous leukemia (CML) patients, treatment with imatinib mesilate that potently inhibits Bcr-Abl tyrosine kinase could be a promising therapeutic approach to CML. Imatinib mesilate was shown to inhibit proliferation of bcr-abl-positive cell lines and suppress the formation of bcr-abl-positive colonies in cells derived from bone marrow of CML patients. This compound induced apoptosis in a variety of bcr-abl-positive cells. Moreover, in vivo data indicated that imatinib mesilate suppress growth and formation of bcr-abl-positive tumors in mice. As the profile expected from the preclinical studies, imatinib mesilate showed impressive hematological and cytogenic responses in the clinical trials, including interferon-alpha-resistant or intolerant patients.