TOXICOLOGICAL EVALUATION OF LEPISTA NUDA (BULL. EX FR.) COOKE MYCELIUM PRODUCED BY AN IN VITRO CULTURE METHODOLOGY

Objective: Considering the interest in L. nuda as a source of ingredients for the development of functional food and nutraceuticals has increased, the objective of this study was to evaluate its general toxicity and possible genotoxic effects in rats to assess its safety.Methods: This study evaluated the safety of L. nuda mycelium by using genotoxicity assays (reverse mutation, chromosomal aberration, and micronuclei tests) and a short-term toxicity test.Results: Our results have indicated that L. nuda mycelium did not significantly increase the number of revertant colonies and chromosomal aberration in both in vitro assays. Furthermore, it did not induce any increase in micronuclei formation in mouse bone marrow.Conclusion: In summary, no mutagenic effects and no evidence of systemic toxicity were found in this safety assessment, and the use of L. nuda mycelia is safe at a dose of 3 g/kg body weight in S-D rats. Using a safety factor of 100, the calculated acceptable daily intake in humans is 30 mg/kg body weight/d

Studies on the Bioactive Components Isolated from the Hot Water Extract of Glechoma hederacea

金錢薄荷(Glechoma hederacea L.)為唇形花科(Lamiaceae)連錢草屬(Glechoma)，為多年生匍匐性草本植物，原產地為歐洲至高加索、北美洲地區，台灣全境低海拔山區、路旁、荒地、陰溼地皆可見其蹤影，分布廣泛，具抗發炎、抗菌及抗氧化等活性。本研究將進行金錢薄荷抽取成份之單離與鑑定。 將金錢薄荷全草以水為溶劑進行加熱迴流萃取，過濾之後經冷凍乾燥及減壓濃縮得到的萃取物利用大孔吸附樹脂層析法(Diaion HP-20)、膠體過濾法(Sephadex LH-20)及逆向ODS填充管柱等過程共分離純化得到8個黃酮類化合物(GF1-GF8)，分別為Apigenin 6,8-di-C-β-glucopyranoside (vicenin-2) (GF1)、Apigenin 6-C-α-arabinopyranosyl-8-C-β -glucopyranoside (isoschaftoside) (GF2)、Luteolin 7-O-β-glucoside (GF3)、Luteolin 7-O-β-glucuronide (GF4)、Apigenin 7-O-β-glucoside (GF5)、Apigenin 7-O-β-glucuronide (GF6)、Luteolin (GF7)和Apigenin (GF8)，均經光譜分析與比對，確認其結構。 定性方面採用高效能液相層析儀進行分析。將金錢薄荷熱水萃取物之指紋圖譜與分離所得8個標準品比對，其滯留時間分別為11.4 min (GF1)、12.8 min (GF2)、15.6 min (GF3)、15.9 min (GF4)、17.5 min (GF5)、18.3 min (GF6)、23.2 min (GF7)及26.7 min (GF8)。Glechoma hederacea L. is a perennial creeping herbal plant of family Lamiaceae, genus Glechoma . It originate in Europe to Caucasus mountain and the region of North America, also growing at roadsides、 wasteland and damp ground in Taiwan. It is widely distributed, and the anti-inflammatory、antibacterial and antioxidant activities have also been reported. In this research, bioactive compounds were isolated and identified for the Glechoma hederacea L.. The hot water extract of dried whole plant of Glechoma hederacea L. was separated by column chromatography on macroporous resin chromatography (Diaion HP-20)、gel filtration (Sephadex LH-20) and reverse phase ODS column to give eight flavonoids (GF1-GF8). The eight compounds were as follows: Apigenin 6,8-di-C-β-glucopyranoside (vicenin-2) (GF1)、Apigenin 6-C-α-arabinopyranosyl-8-C-β -glucopyranoside (isoschaftoside) (GF2)、Luteolin 7-O-β-glucoside (GF3)、Luteolin 7-O-β-glucuronide (GF4)、Apigenin 7-O-β-glucoside (GF5)、Apigenin 7-O-β-glucuronide (GF6)、Luteolin (GF7) and Apigenin (GF8). We established the qualitative analysis of the bioactive components in the Glechoma hederacea L. by HPLC/UV. The chromatogram of the hot water extracts of Glechoma hederacea L. shows eight peaks which were coinside with the standards isolated from it, and their retention time were 11.4 min (GF1)、12.8 min (GF2)、15.6 min (GF3)、15.9 min (GF4)、17.5 min (GF5)、18.3 min (GF6)、23.2 min (GF7) and 26.7 min (GF8)致謝 i 摘要 ii Abstract iii 目次 v 表次 vii 圖次 viii 第一章 緒論 第一節 金錢薄荷簡介 1 第二節 文獻回顧之生物活性 2 第三節 文獻回顧之成份單離 4 第四節 研究目的 7 第二章 金錢薄荷成分之抽取與分離 第一節 金錢薄荷之萃取 8 第二節 金錢薄荷熱水萃取物之成分分離 8 第三章 金錢薄荷化合物結構之研究 第一節Apigenin 6,8-di-C-β-glucopyranoside (GF1)之結構研究 10 第二節Apigenin 6-C-α-arabinopyranosyl-8-C-β -glucopyranoside (isoschaftoside) (GF2)之結構研究 19 第三節Luteolin 7-O-β-glucoside (GF3) 32 第四節Luteolin 7-O-β-glucuronide (GF4) 40 第五節Apigenin 7-O-β-glucoside (GF5)之結構研究 50 第六節Apigenin 7-O-β-glucuronide (GF6)之結構研究 61 第七節Luteolin (GF7)之結構研究 68 第八節Apigenin (GF8)之結構研究 74 第四章 金錢薄荷熱水萃取物成分之定性分析 第一節 高效能液相層析儀及層析條件 79 第二節 樣品溶液製備 79 第三節 實驗結果 80 第五章 結果與討論 第一節 黃酮類之物性 82 第二節 成分之生物活性文獻回顧 83 第六章 結論 86 第七章 實驗部分 儀器與藥品 87 實驗數據 89 參考文獻 9

Safety assessment of HEA‐enriched Cordyceps cicadae mycelia on the central nervous system (CNS), cardiovascular system, and respiratory system in ICR male mice

Abstract Cordyceps cicadae, an entomopathogenic fungus, is a source of traditional Chinese medicine in China. Due to the low yield of wild C. cicadae, artificial cultivation approaches will be needed to meet the increasing market demand. Using bioreactor culture can increase mass production and the abundance of the active component, N6‐(2‐hydroxyethyl)‐adenosine (HEA). Here, we describe a safety assessment for a novel mycelium preparation method. Many studies have confirmed the safety of C. cicadae mycelia. However, the acute safety pharmacology of the C. cicadae enriched with the high HEA (3.90 mg/g) compound has not been evaluated. This study evaluated the central nervous system (CNS), cardiovascular system, and respiratory system in ICR male mice via oral gavage administration. For each requested item, two batches of eight mice tested on a vehicle (0.5% carboxymethyl cellulose, CMC) and C. cicadae mycelia (1,000 mg/kg) were performed. The heart rate at 60 min for the vehicle and C. cicadae mycelium treatment was 700.3 ± 55.4 and 603.0 ± 42.3 bpm, respectively (p = .4279). For echocardiographic analysis, the LV mass of the vehicle and drug treatment was 86.7 ± 6.4 and 80.2 ± 7.7, respectively (p = .0933). In the respiratory test, the tidal volume of the vehicle and drug treatments was 0.11 ± 0.01 and 0.14 ± 0.01 at 60 min, respectively (p = .4262). These results demonstrate that the oral administration of HEA‐enriched C. cicadae mycelia is safe for the CNS, cardiovascular, and respiratory systems

Probiotic Formula Ameliorates Renal Dysfunction Indicators, Glycemic Levels, and Blood Pressure in a Diabetic Nephropathy Mouse Model

One-third of patients with end-stage chronic kidney disease (CKD) experience diabetic nephropathy (DN), which worsens the progression of renal dysfunction. However, preventive measures for DN are lacking. Lactobacillus acidophilus TYCA06, Bifidobacterium longum subsp. infantis BLI-02, and Bifidobacterium bifidum VDD088 probiotic strains have been demonstrated to delay CKD progression. This study evaluated their biological functions to stabilize blood-glucose fluctuations and delay the deterioration of renal function. The db/db mice were used to establish a DN animal model. This was supplemented with 5.125 × 109 CFU/kg/day (high dose) or 1.025 × 109 CFU/kg/day (low dose) mixed with probiotics containing TYCA06, BLI-02, and VDD088 for 8 weeks. Blood urea nitrogen (BUN), serum creatinine, blood glucose, and urine protein were analyzed. Possible mechanisms underlying the alleviation of DN symptoms by probiotic strains were evaluated through in vitro tests. Animal experiments revealed that BUN, serum creatinine, and blood glucose upon probiotic administration were significantly lower than in the control group. The rate of change of urine protein decreased significantly, and blood pressure, glucose tolerance, and renal fibrosis were improved. In vitro testing indicated that TYCA06 and BLI-02 significantly increased acetic acid concentration. TYCA06, BLI-02, and VDD088 were associated with better antioxidation, anti-inflammation, and glucose consumption activities relative to the control. A combination of the probiotics TYCA06, BLI-02, and VDD088 attenuated renal function deterioration and improved blood-glucose fluctuation in a diabetes-induced CKD mouse model

Probiotic Lactobacillus fermentum TSF331, Lactobacillus reuteri TSR332, and Lactobacillus plantarum TSP05 improved liver function and uric acid management-A pilot study.

Metabolic-associated fatty liver disease (MAFLD) is predominantly associated with metabolic disturbances representing aberrant liver function and increased uric acid (UA) levels. Growing evidences have suggested a close relationship between metabolic disturbances and the gut microbiota. A placebo-controlled, double-blinded, randomized clinical trial was therefore conducted to explore the impacts of daily supplements with various combinations of the probiotics, Lactobacillus fermentum TSF331, Lactobacillus reuteri TSR332, and Lactobacillus plantarum TSP05 with a focus on liver function and serum UA levels. Test subjects with abnormal levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and UA were recruited and randomly allocated into six groups. Eighty-two participants successfully completed the 60-day intervention without any dropouts or occurrence of adverse events. The serum AST, ALT, and UA levels were significantly reduced in all treatment groups (P < 0.05). The fecal microbiota analysis revealed the intervention led to an increase in the population of commensal bacteria and a decrease in pathobiont bacteria, especially Bilophila wadsworthia. The in vitro study indicated the probiotic treatments reduced lipid accumulation and inflammatory factor expressions in HepG2 cells, and also promoted UA excretion in Caco-2 cells. The supplementation of multi-strain probiotics (TSF331, TSR332, and TSP05) together can improve liver function and UA management and may have good potential in treating asymptomatic MAFLD. Trial registration. The trial was registered in the US Library of Medicine (clinicaltrials.gov) with the number NCT06183801 on December 28, 2023

Understanding ADC variation by fat content effect using a dual-function MRI phantom

Abstract Background A dual-function phantom designed to quantify the apparent diffusion coefficient (ADC) in different fat contents (FCs) and glass bead densities (GBDs) to simulate the human tissues has not been documented yet. We propose a dual-function phantom to quantify the FC and to measure the ADC at different FCs and different GBDs. Methods A fat-containing diffusion phantom comprised by 30 glass-bead-containing fat-water emulsions consisting of six different FCs (0, 10, 20, 30, 40, and 50%) multiplied by five different GBDs (0, 0.1, 0.25, 0.5, and 1.0 g/50 mL). The FC and ADC were measured by the “iterative decomposition of water and fat with echo asymmetry and least squares estimation-IQ,” IDEAL-IQ, and single-shot echo-planar diffusion-weighted imaging, SS-EP-DWI, sequences, respectively. Linear regression analysis was used to evaluate the relationship among the fat fraction (FF) measured by IDEAL-IQ, GBD, and ADC. Results The ADC was significantly, negatively, and linearly associated with the FF (the linear slope ranged from -0.005 to -0.017, R2 = 0.925 to 0.986, all p < 0.001). The slope of the linear relationship between the ADC and the FF, however, varied among different GBDs (the higher the GBD, the lower the slope). ADCs among emulsions across different GBDs and FFs were overlapped. Emulsions with low GBDs plus high FFs shared a same lower ADC range with those with median or high GBDs plus median or lower FFs. Conclusions A novel dual-function phantom simulating the human tissues allowed to quantify the influence of FC and GBD on ADC. Relevance statement The study developed an innovative dual-function MRI phantom to explore the impact of FC on ADC variation that can affect clinical results. The results revealed the superimposed effect on FF and GBD density on ADC measurements. Key points • A dual-function phantom made of glass bead density (GBD) and fat fraction (FF) emulsion has been developed. • Apparent diffusion coefficient (ADC) values are determined by GBD and FF. • The dual-function phantom showed the mutual ADC addition between FF and GBD. Graphical Abstrac