The different molecular forms of urine neutrophil gelatinase-associated lipocalin present in dogs with urinary diseases

Neutrophil gelatinase-associated lipocalin (NGAL) is a useful biomarker for the early prediction of renal diseases. NGAL may exist as monomer, dimer and/or NGAL/MMP-9 complex forms in humans. In this study, the existence of various forms of NGAL in urine (uNGAL) was determined and whether these forms are related to the different urinary diseases found in dogs is further discussed

Post genomics era for orchid research

Among 300,000 species in angiosperms, Orchidaceae containing 30,000 species is one of the largest families. Almost every habitats on earth have orchid plants successfully colonized, and it indicates that orchids are among the plants with significant ecological and evolutionary importance. So far, four orchid genomes have been sequenced, including Phalaenopsis equestris, Dendrobium catenatum, Dendrobium officinale, and Apostaceae shengen. Here, we review the current progress and the direction of orchid research in the post genomics era. These include the orchid genome evolution, genome mapping (genome-wide association analysis, genetic map, physical map), comparative genomics (especially receptor-like kinase and terpene synthase), secondary metabolomics, and genome editing

Cytotoxic Phenylpropanoids and a New Triterpene, Turformosinic Acid, from Turpinia formosana Nakai

One new phenylpropanoid, turformosin A (1), and one new triterpene, turformosinic acid (2), together with 16 known compounds, were isolated from the stems of Turpinia formosana Nakai. All structures were elucidated on the basis of spectroscopic analysis, including 1D- and 2D-NMR techniques and MS analysis. Selected isolated compounds were evaluated for in vitro cytotoxicity against four human cancer cell lines and antioxidant scavenging effects on DPPH. (-)-(7'S,8'S)-threo-carolignan X (3) exhibited cytotoxicity against Hep2, WiDr, Daoy, and MCF-7 cell lines with ED(50) values of 3.60, 4.45, 6.07, and 13.7 μg/mL, respectively. Turformosin A (1), (-)-(7'S,8'S)- threo-carolignan X (3), methoxyhydroquinone-4-β-D-glucopyranoside (5), and methoxy-hydroquinone-1-β-D-glucopyranoside (6), exhibited similar anti-oxidative activity. Hep2 cells treated with 10 μg/mL of 3 showed elevation of sub-G1 population (from 20% at 8 h to 60% at 48 h), and activation of caspase-9/caspase-3/PARP cascade. Compound 3 induced intrinsic apoptotic pathway in Hep2 cells with dose and time dependence (10 μg/mL for 8 h)

Lessons Learned of NSPO’s Picosatellite Mission: Yamsat - 1A, 1B & 1C

The YamSat is the first developed picosatellite in National Space Program Office’s (NSPO), Taiwan, R.O.C. It is scheduled to flight in the CubeSat launch in 2003. The rapid-prototyping system engineering different from the past formal discipline opens a new satellite development model in NSPO. The YamSat Test Readiness Review Meeting was successfully held in January 2002 and the environmental tests were completed by end March 2002. Besides the breadboard model and engineering test bed to prove of operation concept are built, three YamSats (1A, 1B, & 1C) instead of one are manufactured with slightly different configurations and purposes. The YamSat- 1A is for flight with ambitious and novel R.O.C. made components, including 15 domestic organizations and companies’ participation. The YamSat-1B is basically for backup purpose and demonstration, whereas the YamSat-1C is for amateur communication experiment end-to-end field test, and for public education purpose. This new experience gives fruitful lessons learned and provides low cost space experimentation and education to the next built picosatellites in Taiwan’s universities. Detailed mission and lessons learned are addressed in this paper

BlueBerry Isolate, Pterostilbene, Functions as a Potential Anticancer Stem Cell Agent in Suppressing Irradiation-Mediated Enrichment of Hepatoma Stem Cells

For many malignancies, radiation therapy remains the second option only to surgery in terms of its curative potential. However, radiation-induced tumor cell death is limited by a number of factors, including the adverse response of the tumor microenvironment to the treatment and either intrinsic or acquired mechanisms of evasive resistance, and the existence of cancer stem cells (CSCs). In this study, we demonstrated that using different doses of irradiation led to the enrichment of CD133+ Mahlavu cells using flow cytometric method. Subsequently, CD133+ Mahlavu cells enriched by irradiation were characterized for their stemness gene expression, self-renewal, migration/invasion abilities, and radiation resistance. Having established irradiation-enriched CD133+ Mahlavu cells with CSC properties, we evaluated a phytochemical, pterostilbene (PT), found abundantly in blueberries, against irradiation-enriched CSCs. It was shown that PT treatment dose-dependently reduced the enrichment of CD133+ Mahlavu cells upon irradiation; PT treatment also prevented tumor sphere formation, reduced stemness gene expression, and suppressed invasion and migration abilities as well as increasing apoptosis of CD133+ Mahlavu CSCs. Based on our experimental data, pterostilbene could be used to prevent the enrichment of CD133+ hepatoma CSCs and should be considered for future clinical testing as a combined agent for HCC patients

High-resolution spatial and genomic characterization of coral-associated microbial aggregates in the coral Stylophora pistillata

Bacteria commonly form aggregates in a range of coral species [termed coral-associated microbial aggregates (CAMAs)], although these structures remain poorly characterized despite extensive efforts studying the coral microbiome. Here, we comprehensively characterize CAMAs associated with Stylophora pistillata and quantify their cell abundance. Our analysis reveals that multiple Endozoicomonas phylotypes coexist inside a single CAMA. Nanoscale secondary ion mass spectrometry imaging revealed that the Endozoicomonas cells were enriched with phosphorus, with the elemental compositions of CAMAs different from coral tissues and endosymbiotic Symbiodiniaceae, highlighting a role in sequestering and cycling phosphate between coral holobiont partners. Consensus metagenome--assembled genomes of the two dominant Endozoicomonas phylotypes confirmed their metabolic potential for polyphosphate accumulation along with genomic signatures including type VI secretion systems allowing host association. Our findings provide unprecedented insights into Endozoicomonas-dominated CAMAs and the first direct physiological and genomic linked evidence of their biological role in the coral holobiont

Chronic Kidney Disease, But Not Diabetes, Can Predict 30-Day Outcomes in Patients with ST-Elevation Myocardial Infarction after Primary Percutaneous Coronary Intervention: A Single-Center Experience

Background: Patients with acute coronary syndrome and impaired renal function have been shown to have high mortality. However, there is scarce literature to date addressing the impact of diabetes mellitus (DM) and renal function on clinical outcomes of ST elevation myocardial infarction (STEMI) in Taiwan. Method: This study enrolled 512 STEMI patients who received primary percutaneous coronary intervention. Patients were divided into 4 groups including group 1: patients without DM or CKD (nDM-nCKD); group 2: patients with DM but without CKD (DM-nCKD); group 3: patients with CKD but without DM (nDM-CKD); group 4: patients with DM and CKD (DM-CKD). Patients were also classified into four groups based on their estimated glomerular filtration rates (eGFR): stage 1 (eGFR 90 ml/min/1.73 m 2 , n = 163), stage 2 (eGFR = 89-60 ml/min/1.73 m 2 , n = 171), stage 3 (eGFR = 59-30 ml/min/1.73 m 2 , n = 136), and stage 4 (eGFR < 30 ml/min/1.73 m 2 , n = 42). The complication rates, length of hospital stay, and 30-day outcomes were analyzed. Results: The patients in both the nDM-CKD group and DM-CKD group had higher incidences of hypotension, intra-aortic balloon counterpulsation use, and respiratory failure (p < 0.005). They had significantly longer hospital stay and 30-day mortality rates (p < 0.001). The patients with CKD stage 3 and 4 had longer hospital stay and higher 30-day mortality rates (p < 0.001). However, DM was not an independent factor on the length of hospital stay and 30-day mortality rates. Conclusions: STEMI patients with impaired renal function, but not DM, had significantly longer hospital stay and higher 30-day mortality rates

Women with endometriosis have higher comorbidities: Analysis of domestic data in Taiwan

AbstractEndometriosis, defined by the presence of viable extrauterine endometrial glands and stroma, can grow or bleed cyclically, and possesses characteristics including a destructive, invasive, and metastatic nature. Since endometriosis may result in pelvic inflammation, adhesion, chronic pain, and infertility, and can progress to biologically malignant tumors, it is a long-term major health issue in women of reproductive age. In this review, we analyze the Taiwan domestic research addressing associations between endometriosis and other diseases. Concerning malignant tumors, we identified four studies on the links between endometriosis and ovarian cancer, one on breast cancer, two on endometrial cancer, one on colorectal cancer, and one on other malignancies, as well as one on associations between endometriosis and irritable bowel syndrome, one on links with migraine headache, three on links with pelvic inflammatory diseases, four on links with infertility, four on links with obesity, four on links with chronic liver disease, four on links with rheumatoid arthritis, four on links with chronic renal disease, five on links with diabetes mellitus, and five on links with cardiovascular diseases (hypertension, hyperlipidemia, etc.). The data available to date support that women with endometriosis might be at risk of some chronic illnesses and certain malignancies, although we consider the evidence for some comorbidities to be of low quality, for example, the association between colon cancer and adenomyosis/endometriosis. We still believe that the risk of comorbidity might be higher in women with endometriosis than that we supposed before. More research is needed to determine whether women with endometriosis are really at risk of these comorbidities

Concentration and forms of urine neutrophil gelatinase-associated lipocalin (NGAL) in dogs with urinary diseases

Neutrophil gelatinase-associated lipocalin（NGAL）為隸屬於lipocalin家族之大小約25 kDa分泌性醣蛋白，最初由人類嗜中性球中被發現。許多人醫研究皆指出，腎臟受損後NGAL會於短時間大量表現於血液及尿液中，因此NGAL於早期預測腎衰竭之角色也備受重視。除腎臟來源外，NGAL也會由嗜中性球分泌，有人醫研究發現，下泌尿道感染時尿中NGAL濃度的上升可作為早期偵測下泌尿道感染的指標之一，換句話說，泌尿道感染可能成為NGAL於腎臟疾病應用之干擾因子。NGAL以monomer、dimer及NGAL/MMP-9複合物以上3種形式存在，近期有研究以immunoblotting方式分析發現，腎臟上皮細胞分泌之NGAL以monomer為主，而嗜中性球分泌之型態多以dimer為主。本實驗室已成功製備出可用於偵測犬NGAL之多株抗體且建立ELISA系統，並證明NAGL可成為評估犬之腎衰竭一個有用的生化檢測；然而，白血球數量對犬尿中NGAL濃度及型態之影響仍尚未明瞭。本次實驗目的有二：第一，利用實驗室建立之ELISA系統評估血液及尿中白血球對尿中NGAL的影響；第二，以西方墨點法分析犬尿中NGAL型態，及表現型態是否與腎衰竭及尿中白球有關。由中興大學獸醫教學醫院共收集70隻就診犬之尿液樣本（24隻為腎衰竭病例[RF]，19隻為膿尿病例[PYU]，17隻為腎衰竭同時併發膿尿之病例[RF-PYU]，10隻為非腎衰竭及膿尿之其他疾病病例），以及11隻健康犬之尿液做為健康對照組。由統計分析結果得知，血中白血球數目與尿中NGAL濃度不具相關性（r2 <0.1）；然而，膿尿病例之尿中NGAL濃度則顯著高於對照組（p <0.01）（15.35 ng/mL [IQR 27.66] vs. 3.92 ng/mL [IQR 9.05]）；另外，腎衰竭及腎衰竭同時併發膿尿之病例尿中NGAL濃度顯著高於對照組（RF：23.77 ng/mL [IQR 32.73]、RF-PYU：18.97 ng/mL [IQR 70.32]），但兩組與膿尿病例則無顯著差異。西方墨點法分析所有尿液樣本發現，犬尿中NGAL存在3種型態：monomer（25 kDa）、dimer（50 kDa）及NGAL/MMP-9複合物（150 kDa）。在所有腎衰竭的病例中（RF及RF-PYU），monomer出現的比例顯著高於非腎衰竭病例；而所有膿尿的病例中（PYU及RF-PYU）出現dimer的比例則顯著高於為非膿尿病例。以上結果顯示，犬尿中白血球數目會影響尿中NGAL之濃度；而monomer的出現與腎衰竭有關，dimer的出現則較與尿中白血球有關，藉此有助於辨別犬尿中NGAL來源，以提高尿中NGAL於犬腎衰竭相關臨床應用。Neutrophil gelatinase-associated lipocalin (NGAL), a 25 kDa glycoprotein existing as monomer, homodimer and NGAL/MMP-9 complex, was found highly accumulated in human and mouse proximal tubule during acute tubular necrosis, thus regarded as an early predictor of acute kidney injury (AKI). Recent studies showed that the major molecular form of NGAL secreted by neutrophils is dimeric, whereas the major form secreted by kidney epithelial cells is monomeric. The aim of this study is to evaluate possible factors affecting the concentration and forms of urine NGAL (uNGAL). Urine samples of 70 dogs from Veterinary Medical Teaching Hospital of National Chung Hsing University (24 with renal failure (RF), 19 with pyuria (PYU), 17 with both renal failure and pyuria (RF-PYU), 10 with other diseases) and 11 healthy dogs as control were analyzed by western blotting and home-made ELISA for evaluation of NGAL concentration and forms in urine. Results show no correlation between blood leukocyte counts and uNGAL (r2 <0.1); however, patients with pyuria have higher uNGAL concentration then control (p <0.01). uNGAL in RF and RF-PYU patients were significantly higher than control (p <0.01), but with no significant difference between PYU patients. Three forms of uNGAL were identified in canine urine: monomer, dimer and NGAL/MMP-9 complex, and the rate of monomer in patients with renal failure (RF and RF-PYU) is significantly higher than patients without renal failure (p <0.01) whereas the rate of dimer in patients with pyuria (PYU and RF-PYU) is significantly higher than patients without pyuria(p <0.01). In conclusion, urine leucocyte may cause elevation of uNGAL concentration and could be a confounding factor of uNGAL in clinical use. Moreover, renal failure is associated with the presence of uNGAL monomer whereas dimer is associated with the presence of urine leukocyte, which indicates that the forms of uNGAL may aids in the differentiation of renal failure and pyuria. ii摘要 i Abstract ii 目次 iii 表次 vii 圖次 viii 第一章 緒言 1 第二章 文獻探討 2 第一節 NGAL蛋白之結構特性及介紹 2 一、名稱及起源 2 二、結構及構型 2 三、表現位置 2 第二節 NGAL之調控與其功能 3 一、抑菌功能 3 二、細胞調控因子 4 三、急性炎症蛋白（acute phase protein） 4 第三節 影響NGAL表現之相關疾病及臨床上之應用 5 一、NGAL與腎臟損傷 5 （一）腎臟損傷 5 （二）NGAL於腎損傷之應用 6 二、NGAL與泌尿道感染 8 （一）NGAL與泌尿道感染 8 （二）NGAL於泌尿道感染之應用 8 三、NGAL與胰臟炎 9 （一）胰臟炎 9 （二）NGAL於胰臟炎之應用 9 四、NGAL與心衰竭 9 （一）心衰竭與腎功能受損 9 （二）NGAL於心衰竭之應用 10 五、NGAL與敗血症 10 （一）敗血症 10 （二）NGAL於敗血症及敗血性腎損傷之應用 11 六、NGAL與腫瘤 12 第四節 NGAL之三種構型 12 一、NGAL之三種構型 12 （一）、NGAL之monomer 13 （二）、NGAL之dimer 13 （三）、NGAL/MMP-9之複合體 13 二、不同NGAL型態於臨床診斷上的影響 14 第五節 研究動機 15 第三章 材料與方法 17 第一節 樣本收集 17 一、樣本來源及記錄 17 二、病例分組之標準 17 （一）依臨床數值及診斷之分組 17 （二）依尿中NGAL之單一型態分組 19 三、樣本處理 19 第二節 NGAL於犬尿中之表現 19 一、以酵素連結免疫吸附法（Enzyme-linked immunosorbent assay）測定犬尿中NGAL之濃度 19 （一）重組蛋白Calibrator之訂定 19 （二）樣品處理 19 （三）ELISA 20 二、以西方墨點法（Western blotting）分析NGAL於犬尿中之表現 20 （一）樣品處理 20 （二）SDS（sodiun dodecyl sulfate）-polyacrylamide膠片製作 20 （三）膠體電泳分析 21 （四）西方墨點法 21 第三節 統計學分析 22 一、統計學方法及工具 22 二、統計學研究設計 23 （一）探討尿中出現白血球時，對尿中NGAL濃度的影響。 23 （二）探討尿中白血球是否會影響尿中NGAL於腎衰竭之應用。 23 （三）探討血中白血球數是否會影響尿中NGAL之濃度。 23 （四）尿中NGAL之各種型態存在與否，是否與臨床數值或疾病具關連性。 23 第四章 結果 24 第一節 ELISA結果分析 24 （一）尿中白血球與尿中NGAL濃度之相關性 24 （二）尿中白血球是否會影響尿中NGAL於腎衰竭病例之應用。 27 （三）血中WBC、segment neutrophil、neutrophil數目與尿中NGAL之相關性。 31 第二節 Western blotting分析犬尿液樣本之結果 35 （一）尿中NGAL型態分析 35 （二）尿中NGAL複合體之組成 36 第三節 Western blotting結果之統計學分析 37 （一）尿中不同NGAL型態的存在與尿中NGAL濃度、血液學檢查數值及尿液學檢查結果之相關性。 37 （二）尿中NGAL型態是否可分辨尿中NGAL之可能來源。 39 第五章 討論 41 第六章 結論 46 參考文獻 47 附錄 6

Role of Shortened QTc Dispersion in In-hospital Cardiac Events in Patients Undergoing Percutaneous Coronary Intervention for Acute Coronary Syndrome

QT dispersion (QTD) refers to the difference between maximal and minimal QT values on the electrocardiogram (ECG). QTD values are calculated and corrected with Bazett's formula (corrected QTD = QTcD = QTD/vRR). QTcD increases in patients with acute coronary syndrome (ACS). Recovery of increased QTcD (shortened QTcD) develops after successful revascularization, but prolonged QTcD occurs in certain patients. The aim of this study is to ascertain the clinical significance between shortened and prolonged QTcD groups after percutaneous coronary intervention (PCI). Methods: We retrospectively enrolled 128 patients with ACS who had received PCI. The values of QTcD were measured manually on 12-lead standard ECGs obtained within 3 days before and after PCI (pre-PCI QTcD and post-PCI QTcD). All the patients were divided into 2 groups. The shortened QTcD group was defined as those patients with a decrease in QTcD after PCI and the prolonged QTcD group as those with an increase in QTcD after PCI. The underlying diseases, various clinical classifications and some prognostic factors were taken into comparison and statistical analysis between these 2 groups. Results: The shortened QTcD group showed a significantly higher rate of in-hospital cardiac death (13% vs. 0%, p = 0.006) and a greater pre-PCI QTcD (100.8 ± 39.5 vs. 61.3 ± 24.1 ms, p < 0.001) than the prolonged QTcD group. There was a significantly greater pre-PCI QTcD in patients with cardiac death than those without cardiac death (111.6 ± 38.3 vs. 83.3 ± 38.3 ms, p = 0.027). Furthermore, the patients with in-hospital cardiac death presented with a significantly more frequent occurrence of in-hospital ventricular arrhythmia, compared with those without cardiac death (30.0% vs. 4.0%, p = 0.014). Conclusion: Among the patients with ACS undergoing PCI, directly divided into shortened and prolonged QTcD groups regardless of initial pre-PCI QTcD, the shortened QTcD group showed a higher occurrence of in-hospital cardiac death and a greater pre-PCI QTcD. Shortened QTcD might be 1 risk factor for in-hospital cardiac death