The Quality Improvement of Solid-State Fermentation with Cordyceps militaris by UVB Irradiation

Cordyceps militaris, ljekovita i jestiva gljiva, korištena je za fermentaciju na čvrstoj podlozi od heljde i prirodne riže. Svrha je istraživanja bila ispitati utjecaj ultraljubičastih B (UVB) zraka na udjele vitamina D2 i biološki aktivnih tvari te antioksidacijska svojstva heljde i prirodne riže tijekom fermentacije na čvrstoj podlozi s pomoću C. militaris. Ozračeni uzorci fermentirane heljde i prirodne riže imali su znatno veći udjel vitamina D2, i to od 0-0.3 do 1.18-16.79 μg/g, dok je u svježoj prirodnoj riži udjel vitamina D2 nakon fermentacije bio 16.79 μg/g. Udjeli adenozina, kordicepina i polisaharida u ozračenim sušenim uzorcima fermentiranim s pomoću C. militaris bili su od 0.08 do 11.5 mg/g, odnosno veći nego u svježim uzorcima fermentiranim s pomoću C. militaris (0.07–8.40 mg/g). Uzorci fermentirani s pomoću C. militaris imali su manje vrijednosti EC50 i veći udjel antioksidansa od nefermentiranih uzoraka. Nakon fermentacije na čvrstoj podlozi i zračenja UVB svjetlom, smanjili su se udjeli biološki aktivnih i antioksidacijskih sastojaka te antioksidacijska svojstva uzorka. No, usprkos tome, količina navedenih komponenata bila je dostatna.Cordyceps militaris, a medicinal and edible mushroom, was used to ferment buckwheat and embryo rice by solid-state fermentation (SSF). Our aim was to investigate the effect of ultraviolet B (UVB) light irradiation on the content of vitamin D2 and biologically effective components, and antioxidant properties of buckwheat and embryo rice in SSF with C. militaris. Irradiated samples of buckwheat and embryo rice fermented by C. militaris had significantly increased vitamin D2 content, from 0-0.3 to 1.18-16.79 μg/g, while the increase in fresh embryo rice fermented by C. militaris was up to 16.79 μg/g. The content of adenosine, cordycepin and polysaccharide in irradiated dry samples fermented by C. militaris was 0.08 to 11.15 mg/g, higher than that of the irradiated fresh samples fermented by C. militaris (0.07–8.40 mg/g). Samples fermented by C. militaris had lower EC50 values and higher content of antioxidants than did unfermented samples. When the solid-state fermented sample was irradiated with UVB light, the content of biologically effective and antioxidant components and antioxidant property of sample decreased. However, it still contained enough of these biologically effective and antioxidant components

Activations of Both Extrinsic and Intrinsic Pathways in HCT 116 Human Colorectal Cancer Cells Contribute to Apoptosis through p53-Mediated ATM/Fas Signaling by Emilia sonchifolia Extract, a Folklore Medicinal Plant

Emilia sonchifolia (L.) DC (Compositae), an herbaceous plant found in Taiwan and India, is used as folk medicine. The clinical applications include inflammation, rheumatism, cough, cuts fever, dysentery, analgesic, and antibacteria. The activities of Emilia sonchifolia extract (ESE) on colorectal cancer cell death have not been fully investigated. The purpose of this study explored the induction of apoptosis and its molecular mechanisms in ESE-treated HCT 116 human colorectal cancer cells in vitro. The methanolic ESE was characterized, and γ-humulene was formed as the major constituent (63.86%). ESE induced cell growth inhibition in a concentration- and time-dependent response by MTT assay. Apoptotic cells (DNA fragmentation, an apoptotic catachrestic) were found after ESE treatment by TUNEL assay and DNA gel electrophoresis. Alternatively, ESE stimulated the activities of caspase-3, -8, and -9 and their specific caspase inhibitors protected against ESE-induced cytotoxicity. ESE promoted the mitochondria-dependent and death-receptor-associated protein levels. Also, ESE increased ROS production and upregulated the levels of ATM, p53, and Fas in HCT 116 cells. Strikingly, p53 siRNA reversed ESE-reduced viability involved in p53-mediated ATM/Fas signaling in HCT 116 cells. In summary, our result is the first report suggesting that ESE may be potentially efficacious in the treatment of colorectal cancer

Limits and applications of high-precision coral 230Th dating

珊瑚骨骼中的地球化學指標已廣泛應用於古環境重建的研究。準確解釋珊瑚所紀錄的環境和氣候變化，需仰賴精確的定年技術。前人所提出最好的鈾釷定年技術之不準度為2-3年 (after Edwards et al., 1988)，而我們新發展的方法對於100年以內的珊瑚樣本，可以提供1年或更好的精確度。首先，我們精練了化學方法，在處理程序方面僅產生相當於2-3個月的誤差。非在珊瑚骨骼內之封閉系統中衰變的230Th (230Thnr)含量，即230Th初始值，是限制鈾釷定年準確性與精確性的主要參數。 230Th初始含量可藉由230Th/232Th vs. 234U/232Th等時線圖所得之30Th/232Th初始比值做進一步的估計。台灣南部南灣地區現生微孔珊瑚的230Th/232Th初始值為5.2 ± 1.1 × 10-6 (原子數比，在此之後的比值均是原子數比) ，與海水中溶解相的值 (4.0 ± 0.5 × 10-6) 一致，但高於海水中懸浮微粒物質的比値 (3.0 ± 0.7 × 10-6)。在蘇門答臘群島中的North Pagai，珊瑚230Th/232Th初始值和海水中溶解相的釷同位素比值亦相同。這些結果說明珊瑚中的230Thnr主要來自海水中的溶解相。南灣與蘇門答臘群島South Pagai 的研究顯示，區域性珊瑚骨骼中的230Th/232Th初始值在具有相同的水文環境下，沒有發現有變異性存在。North Pagai的現代標本及 475 年珊瑚化石之間的230Th/232Th初始值在時間上並沒有顯著不同。本研究將已建立好的230Th定年技術對蘇門答臘群島1935年大地震事件做了精確定年且進一步發現其250年的地震週期。古氣候的應用方面，利用西赤道太平洋的珊瑚建立一個精確的年代模式以重建其長期的氣候系統。Abstract Coral skeletal geochemical tracers have been used as proxies to retrieve paleo-conditions. Interpretation of coral-inferred environmental and climatic records relies on accurate dating. Our newly-developed 230Th dating technique offers a precision better than 1 year for corals, younger than 100 years, superior to 2-3 years given by previous methods (after Edwards et al., 1988). To approach this objective, chemistry has been refined and only brings about an equivalent age uncertainty of 2-3 months. The most important parameter limiting precision and accuracy of coral 230Th dating is the content of non-radiogenic 230Th (230Thnr) not due to closed system decay within the coral skeletal matrix, so called the initial 230Th level. It can be evaluated with an initial 230Th/232Th ratio earned from a 230Th/232Th vs. 234U/232Th isochron plot. The initial 230Th/232Th ratio for modern Porites corals, collected from Nanwan, southern Taiwan, is 5.2 ± 1.1 × 10-6 (atomic ratio, hereafter), consistent with a value of 4.0 ± 0.5 × 10-6 in the dissolved fraction of seawater and higher than that of 3.0 ± 0.7 × 10-6 in the suspended particulate matter. The initial 230Th/232Th ratio addressed in Porites corals in North Pagai, Sumatra, also captures the dissolved seawater values. The results indicate that 230Thnr is mainly attributed to the dissolved phase of seawater. Isochron plots for modern and fossil corals from Nanwan and from island South Pagai of Sumatra display little spatial variations of initial 230Th/232Th ratio at local settings. No significant temporal variation is found between initial 230Th/232Th values inferred from modern and 475-year-old corals from island North Pagai, Sumatra. Applications of this well-established 230Th dating technique given in this study include (1) a precise age determination of an earthquake in 1935 AD and (2) a discovery of a 250-year recurrence of giant earthquakes in the Sumatran Islands, and 2 (3) a precise age model for long-term climatic reconstruction with coral tracer records in the western equatorial Pacific Ocean.Table of Contents Page List of Figures III List of Tables V Abstract 1 摘 要 3 Chapter 1 Introduction 4 1.1 Coral chronological methods 4 1.2 U-Th theory 8 1.3 U-Th analytical technique 9 1.4 Constraints on high precision coral 230Th dating and strategy 12 Chapter 2 Methods and chemistry 17 2.1 Sites for seawater and coral samples 17 2.1.1 Nanwan 18 2.1.2 Sumatran Islands 20 2.1.3 Vanuatu 22 2.1.4 Coral subsampling 24 2.2 Experiment 27 2.2.1 Labware and chemicals 27 2.2.2 Coral 27 2.2.3 Seawater 29 2.3 Instrumentation 31 2.3.1 Inductively coupled plasma mass spectrometry (ICP-MS) 31 2.3.2 Bias 33 II 2.3.3 Spectral background and instrumental memory blank 33 Chapter 3 Results 35 3.1 Chemistry refinement and blanks 35 3.2 U-Th data 35 Chapter 4 Discussion 45 4.1 U and δ234U in Nanwan seawater 45 4.2 Th and 230Th/232Th in Nanwan seawater 46 4.3 Uranium and δ234U in corals 51 4.4 Thorium in corals 53 4.5 230Th/232Th ratio and source of 230Thnr in coral 54 4.6 Plot of 230Th age vs. banding age 57 4.7 Temporal and spatial variations of initial 230Th/232Th in Porites 59 4.8 Variability of initial 230Thnr in coral 64 4.9 Applications 65 Chapter 5 Conclusions 70 References 72 致謝 81 Appendix 8

Epigenetic Mechanism in Regulation of Endothelial Function by Disturbed Flow: Induction of DNA Hypermethylation by DNMT1.

There is increasing evidence that epigenetic mechanisms such as changes in DNA methylation and histone modification play an important role in regulating cellular functions in physiological and pathophysiological states. We investigated the effects of hemodynamic force disturbance, one of the risk factors for atherogenesis, on DNA methylation in HUVECs and rat carotid arteries. Our results demonstrated that athero-prone oscillatory shear flow (OS) without a clear direction induces DNA hypermethylation in comparison to the athero-protective pulsatile shear flow (PS) with a definite direction. Furthermore, OS increases the expression and nuclear translocation of DNA methyltransferase 1 (DNMT1), which is a major maintenance DNA methyltransferase that adds methyl groups to hemi-methylated DNA to repress gene expression. Pharmacological inhibition of DNMT1 by 5-Aza-2'-deoxycytidine abolished the OS-induced DNA hypermethylation. In vivo experiments also showed increases of DNMT1 expression and DNA methylation in the partially-ligated rat carotid arteries where the shear flow is disturbed. These in vitro and in vivo findings have provided novel evidence of the differential regulation of DNA methylation by different hemodynamic forces acting on vascular endothelium and identified DNMT1 as a key protein that governs the epigenetic changes in response to the pathophysiological stimuli due to disturbed flow

Epigenetic Mechanism in Regulation of Endothelial Function by Disturbed Flow: Induction of DNA Hypermethylation by DNMT1.

There is increasing evidence that epigenetic mechanisms such as changes in DNA methylation and histone modification play an important role in regulating cellular functions in physiological and pathophysiological states. We investigated the effects of hemodynamic force disturbance, one of the risk factors for atherogenesis, on DNA methylation in HUVECs and rat carotid arteries. Our results demonstrated that athero-prone oscillatory shear flow (OS) without a clear direction induces DNA hypermethylation in comparison to the athero-protective pulsatile shear flow (PS) with a definite direction. Furthermore, OS increases the expression and nuclear translocation of DNA methyltransferase 1 (DNMT1), which is a major maintenance DNA methyltransferase that adds methyl groups to hemi-methylated DNA to repress gene expression. Pharmacological inhibition of DNMT1 by 5-Aza-2'-deoxycytidine abolished the OS-induced DNA hypermethylation. In vivo experiments also showed increases of DNMT1 expression and DNA methylation in the partially-ligated rat carotid arteries where the shear flow is disturbed. These in vitro and in vivo findings have provided novel evidence of the differential regulation of DNA methylation by different hemodynamic forces acting on vascular endothelium and identified DNMT1 as a key protein that governs the epigenetic changes in response to the pathophysiological stimuli due to disturbed flow

Differences in the healthcare needs of older adults attending primary health centers in urban and rural areas of Taiwan

Abstract Purpose As the older adult population in Taiwan continues to increase, primary health centers (PHCs) play a crucial role in geriatric care. This study explored the differences in the PHC experiences and usage needs of older adults in urban versus rural areas. Methods A qualitative study was conducted. Twenty-one older adults were recruited from PHCs in northern, central, southern, and eastern Taiwan. Semistructured interviews were used to collect data, and the interview guidelines included their reasons for visiting PHCs, the health-care services they received, their evaluation of the services, and the advantages and disadvantages of these centers. The data were analyzed using thematic content analysis. Results The PHC usage needs of older adults in urban areas differ from those of older adults in rural areas in the following 3 aspects: (1) Medical services: older adults in rural areas demand clinics specializing in various medical domains, mobile and home medical care, and case management, whereas those in urban areas demand mobile health examinations. (2) Environment and transportation: older adults in rural areas demand diverse medical equipment, shuttle services, and accessible facilities, whereas those in urban areas demand recreational facilities. (3) Active aging: older adults in rural areas demand health education courses, and those in urban areas demand diverse senior citizen courses as well as opportunities to volunteer and build interpersonal relationships. Conclusion The older adults in urban and rural areas had different PHC usage needs. The older adults in rural areas generally focused on medical care and transportation services in PHCs, whereas those in urban areas generally focused on health promotion as a means of social participation and active aging

Antiinflammatory Activity of Gynura bicolor (紅鳳菜 Hóng Fèng Cài) Ether Extract Through Inhibits Nuclear Factor Kappa B Activation

This study investigated effects of the Gynura bicolor (Roxb. and Willd.) DC. ether extract (GBEE) on nitric oxide (NO) and prostaglandin (PG)E2 production on the lipopolysaccharide (LPS)-induced inflammatory response in RAW 264.7 cells. A composition analysis of GBEE showed that the major compounds were b-carotene, chlorophyll, and quercetin, respectively. Furthermore, NO and PGE2 levels of 120 μg/ml GBEE-treated cells were 70% and 9.8%, respectively, than those of cells treated with LPS alone. Immunoblots assays showed that the GBEE dose-dependently suppressed LPS-induced inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 protein levels. The GBEE significantly decreased cytosolic phosphorylated (p)-IκBa and nuclear p65 protein expressions. Electrophoresis mobility shift assays indicated that the GBEE effectively inhibited nuclear factor kappa B (NF-κB) activation induced by LPS. These results support a role of the GBEE in suppressing activation of NF-κB to inhibit NO and PGE2 production in the LPS-induced inflammatory response by RAW 264.7 cells

Opto Field-Effect Transistors for Detecting Quercetin–Cu²⁺ Complex

In this study, we explored the potential of applying biosensors based on silicon nanowire field-effect transistors (bio–NWFETs) as molecular absorption sensors. Using quercetin and Copper (Cu2+) ion as an example, we demonstrated the use of an opto–FET approach for the detection of molecular interactions. We found that photons with wavelengths of 450 nm were absorbed by the molecular complex, with the absorbance level depending on the Cu2+ concentration. Quantitative detection of the molecular absorption of metal complexes was performed for Cu2+ concentrations ranging between 0.1 μM and 100 μM, in which the photon response increased linearly with the copper concentration under optimized bias parameters. Our opto–FET approach showed an improved absorbance compared with that of a commercial ultraviolet-visible spectrophotometry

Primary Small Cell Carcinoma of the Stomach Successfully Treated With Cisplatin and Etoposide

We report a 44-year-old man with primary gastric small cell carcinoma who showed a remarkable response to chemotherapy specific for pulmonary small cell carcinoma. The patient had been admitted to another local hospital because of intermittent epigastralgia. An upper gastrointestinal examination there revealed an ulcerative tumor, 5 cm in diameter, on the lesser curvature side of the cardia, and endoscopic biopsy reported adenocarcinoma. Computed tomography revealed a mass over the lesser curvature of the stomach and some enlarged regional lymph nodes. Radical total gastrectomy, lymph node dissection, Roux-en-Y esophagojejunostomy and splenectomy were performed at our hospital. Pathology revealed gastric mucosa infiltrated by small-sized tumor cells with scanty cytoplasm and hyperchromatic nuclei. Immunohisto- chemically, the tumor cells were positive for synaptophysin, chromogranin A, and CD56. Primary gastric small cell carcinoma was diagnosed. The postoperative course, complicated by shock due to bleeding, wound infection and intra-abdominal abscess, took more than 2 months to resolve. Follow-up computed tomography showed tumor recurrence with multiple enlarged lymph nodes in the aortocaval region and hepatic hilum. The patient received palliative chemotherapy consisting of cisplatin 80 mg/m2 on day 1 and etoposide 80 mg/m2 on days 1–3 every 28 days, and had partial response to the chemotherapy, with a progression-free survival of 10 months. Chemotherapy with cisplatin and etoposide used for small cell carcinoma of the lung is a good treatment for gastric small cell carcinoma