2023 - Vol. 6
Adenosine Acts as an Active Antiplatelet Constituent in Strawberries (Fragaria × ananassa) | Vol.6, No.1, p.27-32 |
---|---|
Natsuki Ichihara , Shiho Maekawa , Nanako Ogawa , Aiko Yamada , Tomoka Nagasato , Ikuro Maruyama , Kazuyoshi Sone , Masaharu Yasuda , Kenji Matsushita , Chihiro Ito , Yoshiaki Takaya | |
Received: January 20, 2023 | |
Accepted: February 04, 2023 | |
Released: March 01, 2023 | |
Abstract | Full Text PDF[1M] |
Many foods have been reported to be effective against thrombosis, but most of them may be difficult to ingest in effective doses to prevent the disease. In this study, we focused on strawberries as one of the foods reported to have antithrombotic activities because they are highly palatable to many people and have several intake methods (e.g., raw fruits, juices, jams, etc.). In this study, strawberry ‘Minomusume’ was selected as a material to explore antiplatelet active compounds. The activity-guided fractionation of the strawberry extract resulted in the active compound being adenosine. Furthermore, we investigated the correlation between adenosine content and the antiplatelet activity of strawberries. As a result, as for the cultivars whose adenosine was detected, significant positive correlation was observed.
Elucidation of the Usefulness of Glypican-3, a Hepatocellular Carcinoma Biomarker, with the Fully Automated LUMIPULSE® System | Vol.6, No.1, p.21-26 |
---|---|
Hisashi Nojima , Tomonori Nishii , Kumiko Iida , Paul Franck Adeyissimi Adjou Moumouni , Katsumi Aoyagi , Shintaro Yagi | |
Received: November 15, 2022 | |
Accepted: January 24, 2023 | |
Released: February 15, 2023 | |
Abstract | Full Text PDF[1M] |
Background: Glypican-3 (GPC3) is expressed in most of hepatocellular carcinoma (HCC) and GPC3 immunohistochemical staining is widely used in the clinical setting, but it has not been recommended as a blood biomarker, mainly due to its heterogeneous nature and the lack of established assay system. Here, we developed and evaluated the basic performance of fully automated GPC3 immunoassay kits which detect the full-length or the N-terminal fragments. We analyzed the molecular forms of GPC3 in HCC serum and evaluated the diagnostic performance of GPC3 and other biomarkers. Methods: We examined the analytical performance of the GPC3 kits. Then, the automated GPC3 assays were compared with an established ELISA kit. Afterwards, we determined the clinical cutoff of GPC3 and compared its diagnostic performance to alpha-fetoprotein (AFP) and protein induced by vitamin K absence or antagonist-II (PIVKA-II) using 180 serum samples from clinically diagnosed patients. Results: GPC3 assays showed good analytical performance. The level of GPC3 in HCC was higher than recorded in healthy or other liver diseases’ sera. The AUC of GPC3 was 0.90, whereas the AUCs of AFP and PIVKA-II were 0.89 and 0.76, respectively. Conclusion: Automated GPC3 assays with stable performance against GPC3 in screening HCC have been established and the diagnostic accuracy of GPC3 was as good as AFP.
Dose-Related Difference and Involvement of Tyrosine Phosphorylation in Methylmercury-Induced Morphological Change in Cultured Astrocytes Obtained from Rat Cerebral Hemisphere | Vol.6, No.1, p.16-20 |
---|---|
Tatsumi Adachi , Hideaki Shimizu , Kohei Kawasumi , Yuka Suzuki , Tomoharu Hayashi , Ryota Tsurusaki , Yosuke Watanabe , Mitsunori Iwasaki , Yasuhide Yanagisawa , Ichiro Yasuda | |
Received: January 10, 2023 | |
Accepted: January 17, 2023 | |
Released: February 01, 2023 | |
Abstract | Full Text PDF[2M] |
Astrocytes obtained from the cerebral hemisphere were maintained in serum-free defined medium containing dibutyryl cyclic AMP (dbcAMP) for 4 h, at which time almost all astrocytes showed a process-bearing stellate shape. They were then exposed to methylmercury (MeHg) at 1–3 μM or solvent alone (control) for up to 24 h. Astrocytes showed a flat polygonal shape after MeHg exposure at 3 µM for 1 h or at 2 µM for 3 h, whereas the shape was not affected after exposure at less than 1 µM for 24 h. Mercury accumulation levels were similar after MeHg exposure at 3 µM for 1 h and at 2 µM for 3 h, while the level after exposure at 1 µM did not reach those levels. The shape of astrocytes exposed to MeHg at 2 µM returned to stellate from polygonal after at least 24 h, although it remained polygonal in astrocytes exposed to MeHg at 3 µM. The viable cell number was significantly lower than in the control culture only in the culture exposed to MeHg at 3 µM for 24 h. In addition, genistein, a tyrosine kinase inhibitor, inhibited the morphological changes (spreading) induced by MeHg at 2–3 µM. These results suggest that a dose-dependent difference is observed in shape changes and cell death caused by MeHg in astrocytes, and that the responses to MeHg correlate to its accumulation levels, especially for the first few hours. They also suggest that tyrosine phosphorylation may play an important role in MeHg-induced spreading in astrocytes.
Evaluation of Recombinant Cascade Reagent PyroSmart NextGen® and Limulus Amebocyte Lysate Equivalency in a Plate and Tube Reader for Bacterial Endotoxins Testing | Vol.6, No.1, p.11-15 |
---|---|
Madeline Kelley , Ingrid Stevens , Nathaniel Marchessault , Junko Akiyoshi , Edwin G. Jahngen | |
Received: November 19, 2022 | |
Accepted: December 08, 2022 | |
Released: January 18, 2023 | |
Abstract | Full Text PDF[1M] |
PyroSmart NextGen® is a recombinant cascade reagent (rCR) for the detection and quantification of bacterial endotoxins developed using cloned genes derived from the Limulus polyphemus horseshoe crab genome. Requirements for use of this alternative reagent include analysis of analytical performance, method suitability, and test result equivalency to Limulus amoebocyte lysate (LAL) reagents used in the compendial Bacterial Endotoxins Test (BET). The plate reader evaluation has been expanded to address two long-standing user preferences, with the inclusion of the tube reader method increasing the sensitivity of endotoxin detection from 0.005 EU/mL to 0.001 EU/mL. The utilization of PyroSmart NextGen® with the two different instrument types also allows for a more comprehensive equivalency analysis. Furthermore, the comparison results demonstrate that PyroSmart NextGen® detects equivalent levels of autochthonous endotoxin in water samples when compared to LAL reagents. Overall, this study provides the first large-scale example of equivalency analysis utilizing a robust rCR and has verified that PyroSmart NextGen® meets the expectations for alternative reagents.
Inhibitory Effects of Olive-Derived Phytochemicals on SARS-CoV-2 3C-Like Protease (3CLpro) | Vol.6, No.1, p.8-10 |
---|---|
Yoshimitsu Kiriyama , Hiromi Nochi | |
Received: November 22, 2022 | |
Accepted: December 22, 2022 | |
Released: January 12, 2023 | |
Abstract | Full Text PDF[761K] |
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the virus that caused the global epidemic of COVID-19. 3C-like protease (3CLpro), called the main protease (Mpro), cleaves 11 sites in the long-stranded protein synthesized from viral genomic RNA. After the cleavage of the long-stranded protein, proteins such as RNA-dependent RNA polymerase are cut off and activated. Therefore, 3CLpro is crucial for SARS-CoV-2 replication and is the target of drugs against SARS-CoV-2. Polyphenols and triterpenoids in olives can bind to SARS-CoV-2 3CLpro in in silico and computational studies. However, not all phytochemicals have been shown to inhibit SARS-CoV-2 3CLpro. Here, we examined the inhibitory effects of olive-derived phytochemicals on SARS-CoV-2 3CLpro. Among these phytochemicals, luteolin and oleanolic acid inhibited the 3CLpro activity by about 40%. The actual enzyme inhibitory action of oleanolic acid has not been examined. Thus, we examined the dose response in the inhibition of 3CLpro by oleanolic acid. Oleanolic acid significantly inhibited the enzymatic activity of 3CLpro at concentrations above 50 μM, and it may be a potent molecule that inhibit SARS-CoV-2 replication via 3CLpro inhibition.
Effects of Hochuekkito on Lenvatinib-Induced Fatigue in Mice | Vol.6, No.1, p.1-7 |
---|---|
Jinyang Xu , Ikuo Nakamura , Makoto Sudo , Satoshi Noda , Naoki Fujitsuka , Sachiko Mogami , Keiko Mitani , Masaharu Tada , Yasuhiro Fujimoto , Tomohiro Terada , Seiko Hirono , Etsuro Hatano | |
Received: November 23, 2022 | |
Accepted: December 08, 2022 | |
Released: January 10, 2023 | |
Abstract | Full Text PDF[1M] |
Lenvatinib has been approved for treating various cancers; however, it exerts numerous adverse effects. Hochuekkito, a Japanese Kampo medicine, can alleviate these adverse effects. Here, we aimed to evaluate the effect of Hochuekkito on lenvatinib-induced chronic fatigue in a murine model. BALB/C mice were fed a control diet or a diet supplemented with 1.5% Hochuekkito for six weeks. On days 15–42, the mice were intraperitoneally injected with dimethyl sulfoxide or lenvatinib. Accordingly, the mice were divided into control/dimethyl sulfoxide, control/lenvatinib, 1.5% Hochuekkito/dimethyl sulfoxide, and 1.5% Hochuekkito/lenvatinib groups. Body weight and food intake were recorded daily. Nesting tests were performed once a week, and the serum interleukin-6 (IL-6) concentration was measured. Liver drug-metabolizing enzyme, CYP3A4, breast cancer resistance protein (BCRP), and P-glycoprotein (P-gp) levels were determined. The serum lenvatinib concentration and CYP3A4, BCRP, and P-gp levels did not differ significantly between the control/lenvatinib and 1.5% Hochuekkito/lenvatinib groups. The usage rate of nesting material on day 42 was higher in the 1.5% Hochuekkito/lenvatinib group than in the control/lenvatinib group (p < 0.05). The serum IL-6 level was lower in the control/dimethyl sulfoxide and 1.5% Hochuekkito/lenvatinib groups than in the control/lenvatinib group (p < 0.05). Overall, Hochuekkito may alleviate lenvatinib-induced fatigue through IL-6 inhibition.