Cucurbitane-type triterpenoids from the vines of Momordica charantia and their anti-inflammatory, cytotoxic, and antidiabetic activity.

Phytochemical investigation of the ethanol extract from wild Momordica charantia vines has resulted in isolation of seven cucurbitane-type triterpenoids, including six undescribed compounds, kuguaovins H‒M, and the known compound, momordicoside K. The structures of the isolated compounds were elucidated on the basis of spectroscopic analyses, including 1D and 2D NMR, and MS experiments. The chemical structure of momordicoside K was determined for the first time by X-ray crystallographic analysis and its absolute configuration assigned. The cytotoxicity against four human tumor cell lines and anti-inflammatory activities on LPS-stimulated RAW264.7 macrophages were evaluated. Of the isolates, kaguaovin L exhibited potential cytotoxicity against MCF-7, HEp-2, Hep-G2, and WiDr cancer cell lines and showed moderate anti-NO production activity. In addition, kuguaovins H and J also showed the stimulatory effect of GLP-1 secretion on the murine intestinal secretin tumor cell line (STC-1).

Cucurbitane-Type Triterpenoids from the Vines of Momordica charantia and Their Anti-inflammatory Activities.

Seven new cucurbitane-type triterpenoids, kuguaovins A-G (1-7), and five known ones were isolated from the rattans of wild Momordica charantia. Their structures were established by spectroscopic data analyses, including 1D and 2D NMR, IR, and MS techniques. The absolute configurations of the cucurbitanes were determined from NOESY data and partially by X-ray crystallographic analysis. In pharmacological studies, compounds 1-7 and 9-12 exhibited weak anti-inflammatory effects (IC50 = 15-35 µM), based on an anti-NO production assay.

Development of analytical method of melamine in placenta from pregnant women by isotope-dilution liquid chromatography/tandem mass spectrometry.

RATIONALE: Melamine is ubiquitously present in our daily life. It has a known effect on the kidneys, but it may also adversely affect the reproduction system. We have developed an analytical method for measuring melamine levels in maternal placenta and correlated these levels with melamine concentrations in urine, a necessary step in finding out if melamine might cross the placenta and enter the circulation of the fetus. METHODS: We used liquid-liquid extraction, clean up by solid-phase extraction (SPE), and isotope-dilution liquid chromatography/tandem mass spectrometry (LC/MS/MS) to measure melamine in placenta specimens. The results of this method were assessed for linearity, limits of quantitation (LOQs), and intra- and inter-assay precision as well as accuracy, matrix effect, and recovery rate. RESULTS: Calibration curves indicated good linearity (r >0.995) over concentrations ranging from 5 to 500 ng/mL in placenta specimens, intra- and inter-assay precision from 0.89% to 27.07%, and accuracy from 92.4% to123.5%. Recovery ranged from 63.9 to 83.9%, and the LOQ was 5 ng/mL in placenta (0.2 g). Placental melamine levels ranged from 7.87 to19.64 ng/mL, all detectable (n = 8). Pregnant women with higher levels of urinary melamine had higher placenta melamine levels than those with non-detectable urinary melamine, though the results were not significantly different (p = 0.149, n = 4 in each group). CONCLUSIONS: The results of this study suggest that pregnant women were exposed to low doses of melamine in their daily lives as measured in urine samples and placenta specimens. It is unclear whether placenta melamine concentrations can better represent long-term exposure than urine or whether melamine in the uterus can enter the fetus via this route.

The First Naked Bismuth-Chalcogen Metal Carbonyl Clusters: Extraordinary Nucleophilicity of the Bi Atom and Semiconducting Characteristics.

Mixed bismuth-chalcogen-iron clusters [{EFe3(CO)9}Bi]- [E = Te (1a) or Se (1b)] were produced via the reduction of BiCl3 with [EFe3(CO)9]2- under mild conditions. X-ray analysis showed that both clusters 1a and 1b had a square-pyramidal geometry, where the naked Bi and chalcogen both adopted a distorted trigonal-pyramidal configuration with a stereoactive lone pair. Complexes 1a and 1b can be further functionalized by methylation and metalation, which permits the nucleophilicity of the 6s/5s and 6s/4s lone pairs to be compared. In the metalation, the 6s pair of the Bi atom in 1a and 1b had an extraordinary nucleophilicity toward the unsaturated Cr(CO)5 fragment, even in the presence of the more chemically active 5s or 4s pair, whereas in the case of methylation, only the 4s pair of Se could be selectively alkylated. Upon oxidation of 1a and 1b with suitable oxidizing agents, NaBiO3 or K2SeO3, Bi-E bonded tetrahedral complexes [{EFe2(CO)6}Bi]- [E = Te (4a) or Se (4b)] were formed by the elimination of one Fe(CO)3 vertex. X-ray photoelectron spectroscopy, X-ray absorption near-edge structure, and density functional theory (DFT) calculations showed that all of the Bi atoms in these complexes had oxidation states close to +1. Due to the electropositive character of the Bi atom, pronounced induced Bi···E inter- and intramolecular interactions were evident in 1a (1b), 4a (4b), and the metalated 3a (3b), where their linear-like ···Bi···E··· or zigzag-like ···Bi-E··· (E = Te or Se) chain or the Bi···E···E···Bi (E = Te or Se) dimeric chain can further expand into the two-dimensional network via nonclassical C-H···O(carbonyl) interactions, supported by noncovalent interaction index and DFT calculations. These positively charged Bi-induced Bi···E (E = Te or Se) and carbonyl-aided weak interactions can facilitate efficient electron transport within these ternary Bi-E-Fe or quaternary Bi-E-Fe-Cr cluster-based frameworks, resulting in semiconducting behavior with surprising ultranarrow energy gaps of 1.01-1.21 eV.

Hepatitis C virus core protein potentiates proangiogenic activity of hepatocellular carcinoma cells.

Increased angiogenic activity has been demonstrated in hepatitis C virus (HCV)-related hepatocellular carcinoma (HCC), but the mechanism was unclear. To study the role of HCV core protein, we used tube formation and Matrigel plug assays to assess the proangiogenic activity of an HCC cell line, HuH7, and 2 of its stable clones-HuH7-core-high and HuH7-core-low, with high and low HCV core protein expression, respectively. In both assays, HuH7-core-high and HuH7-core-low cells dose-dependently induced stronger angiogenesis than control cells. HuH7 cells with HCV core protein expression showed increased mRNA and protein expression of vascular endothelial growth factor (VEGF). VEGF inhibition by bevacizumab reduced the proangiogenic activity of HuH7-core-high cells. The promotor region of VEGF contains the binding site of activator protein-1 (AP-1). Compared with controls, HuH7-core-high cells had an increased AP-1 activity and nuclear localization of phospho-c-jun. AP-1 inhibition using either RNA knockdown or AP-1 inhibitors reduced the VEGF mRNA expression and the proangiogenic activity of HuH7-core-high cells. Among 131 tissue samples from HCC patients, HCV-related HCC revealed stronger VEGF expression than did hepatitis B virus-related HCC. In conclusion, increased VEGF expression through AP-1 activation is a crucial mechanism underlying the proangiogenic activity of the HCV core protein in HCC cells.

Semiconducting Coordination Polymers Based on the Predesigned Ternary Te-Fe-Cu Carbonyl Cluster and Conjugation-Interrupted Dipyridyl Linkers.

A series of semiconducting cluster-incorporated Cu-based coordination polymers, namely, 1D zigzag polymers [{TeFe3 (CO)9 Cu2 }(L)]n (L=1,2-bis(4-pyridyl)ethane (bpea), 1; L=1,2-bis(4-pyridyl)ethylene (bpee), 5), 2D honeycomb-like polymers [{TeFe3 (CO)9 Cu}Cu(L)2.5 ]n (L=bpea, 2; L=bpee, 6), and 2D wave-like cation-anion polymer [{Cu2 (L)4 }({TeFe3 (CO)9 Cu}2 (L))]n (L=1,3-bis(4-pyridyl)propane (bpp), 4), as well as the macrocycle [{TeFe3 (CO)9 Cu2 }2 (bpp)2 ] (3) have been quantitatively synthesized via the liquid-assisted grinding from the pre-designed cluster [TeFe3 (CO)9 Cu2 (MeCN)2 ] with conjugated or conjugation-interrupted dipyridyl linkers. Notably, the most conjugation-interrupted bpp-bridged polymer 4 exhibited extraordinary semiconducting characteristics with an ultra-narrow bandgap of 1.43 eV and a DC conductivity of 1.5×10-2 Ω-1  cm-1 , which violates our knowledge, mainly attributed to the through-space electron transport via non-classical C-H⋅⋅⋅O(carbonyl) hydrogen bonds and aromatic C-H⋅⋅⋅π interactions. The incorporated Te-Fe-CO anions can not only provide numerous possibilities for secondary interactions within these Cu-based polymers but also serve as a redox-active coordination ligand to promote their conductivities. The intriguing structure-property relationships were studied by X-ray and DFT analyses and further demonstrated by significant change in the oxidation state of Cu atoms by XPS and Cu K-edge XANES.

Clinical Decision Support System for Diabetes Based on Ontology Reasoning and TOPSIS Analysis.

Introduction: Although a number of researchers have considered the positive potential of Clinical Decision Support System (CDSS), they did not consider that patients' attitude which leads to active treatment strategies or HbA1c targets. Materials and Methods: We adopted the American Diabetes Association (ADA) and the European Association for the Study of Diabetes (EASD) published to propose an HbA1c target and antidiabetic medication recommendation system for patients. Based on the antidiabetic medication profiles, which were presented by the American Association of Clinical Endocrinologists (AACE) and American College of Endocrinology (ACE), we use TOPSIS to calculate the ranking of antidiabetic medications. Results: The endocrinologist set up ten virtual patients' medical data to evaluate a decision support system. The system indicates that the CDSS performs well and is useful to 87%, and the recommendation system is suitable for outpatients. The evaluation results of the antidiabetic medications show that the system has 85% satisfaction degree which can assist clinicians to manage T2DM while selecting antidiabetic medications. Conclusions: In addition to aiding doctors' clinical diagnosis, the system not only can serve as a guide for specialty physicians but also can help nonspecialty doctors and young doctors with their drug prescriptions.

Iron carbonyl cluster-incorporated Cu(I) NHC complexes in homocoupling of arylboronic acids: an effective [TeFe3(CO)9](2-) ligand.

A new type of TeFe3(CO)9-incorporated dicopper NHC complex was obtained directly from one-pot reactions. By the introduction of the cluster anion [TeFe3(CO)9](2-) and NHCs as the ligands, these di-Cu(i)-based complexes exhibited pronounced catalytic activities toward the homocoupling of arylboronic acids with low Cu loadings and high yields (up to 98%).

High Serum Transforming Growth Factor-ß1 Levels Predict Outcome in Hepatocellular Carcinoma Patients Treated with Sorafenib.

BACKGROUND: The TGF-ß signaling pathway is crucial in the progression and metastasis of malignancies. We investigated whether the serum TGF-ß1 level was related to the outcomes of patients treated with sorafenib for advanced hepatocellular carcinoma (HCC). EXPERIMENTAL DESIGN: We selected patients who had received sorafenib-containing regimens as first-line therapy for advanced HCC between 2007 and 2012. Serum TGF-ß1 levels were measured and correlated with the treatment outcomes. The expression TGF-ß1 and the sensitivity to sorafenib were examined in HCC cell lines. RESULTS: Ninety-one patients were included; 62 (68%) were hepatitis B virus surface antigen (+), and 11 (12%) were anti-hepatitis C virus (+). High (≥ median) pretreatment serum TGF-ß1 levels (median 13.7 ng/mL; range, 3.0-41.8) were associated with high α-fetoprotein levels, but not with age, gender, or disease stage. Patients with high pretreatment serum TGF-ß1 levels exhibited significantly shorter progression-free survival (median, 2.5 vs. 4.3 months; P = 0.022) and overall survival (median 5.6 vs. 11.6 months; P = 0.029) than did patients with low serum TGF-ß1 levels. Compared with pretreatment levels, the serum TGF-ß1 levels were significantly increased at disease progression (n = 29, P = 0.010). In preclinical models of HCC, higher TGF-ß1 expression levels were associated with poorer sensitivity to sorafenib. CONCLUSIONS: High pretreatment serum TGF-ß1 levels were associated with poor prognoses, and increased serum TGF-ß1 levels were associated with the disease progression of advanced HCC patients. TGF-ß pathway may be explored as a therapeutic target for advanced HCC.

Construction of a clinical decision support system for undergoing surgery based on domain ontology and rules reasoning.

OBJECTIVE: To construct a clinical decision support system (CDSS) for undergoing surgery based on domain ontology and rules reasoning in the setting of hospitalized diabetic patients. MATERIALS AND METHODS: The ontology was created with a modified ontology development method, including specification and conceptualization, formalization, implementation, and evaluation and maintenance. The Protégé-Web Ontology Language editor was used to implement the ontology. Embedded clinical knowledge was elicited to complement the domain ontology with formal concept analysis. The decision rules were translated into JENA format, which JENA can use to infer recommendations based on patient clinical situations. RESULTS: The ontology includes 31 classes and 13 properties, plus 38 JENA rules that were built to generate recommendations. The evaluation studies confirmed the correctness of the ontology, acceptance of recommendations, satisfaction with the system, and usefulness of the ontology for glycemic management of diabetic patients undergoing surgery, especially for domain experts. CONCLUSIONS: The contribution of this research is to set up an evidence-based hybrid ontology and an evaluation method for CDSS. The system can help clinicians to achieve inpatient glycemic control in diabetic patients undergoing surgery while avoiding hypoglycemia.

The germline BIM deletion polymorphism is not associated with the treatment efficacy of sorafenib in patients with advanced hepatocellular carcinoma.

OBJECTIVES: A germline BIM deletion polymorphism has been proposed to predict a poor treatment efficacy of certain kinase inhibitors. The current study aimed to explore whether the BIM deletion polymorphism predicts the treatment efficacy of sorafenib for advanced hepatocellular carcinoma (HCC). METHODS: All patients who were enrolled in clinical trials to receive sorafenib-containing regimens as first-line therapy for advanced HCC and consented to providing peripheral blood samples were included. Polymerase chain reaction followed by gel electrophoresis was used to detect the germline BIM deletion polymorphism. RESULTS: A total of 89 patients were enrolled; 69 (77%) patients had chronic hepatitis B infection, and 18 (20%) had chronic hepatitis C infection. The heterozygous BIM deletion polymorphism was identified in 9 (10%) patients. Patients with and without the BIM deletion polymorphism had similar response rates (11 vs. 6%) and disease control rates (56 vs. 61%). The time to progression, progression-free survival, and overall survival were similar between patients with and without the BIM deletion polymorphism. After adjusting for basic clinicopathologic variables and treatment regimens, the BIM polymorphism still could not predict treatment outcomes. CONCLUSIONS: The BIM deletion polymorphism was not associated with the treatment efficacy of sorafenib for advanced HCC.

Trigonal-bipyramidal and square-pyramidal chromium-manganese chalcogenide clusters, [E2CrMn2(CO)n](2-) (E=S, Se, Te; n=9, 10): synthesis, electrochemistry, UV/Vis absorption, and computational studies.

The reactions of E powder (E=S, Se) with a mixture of Cr(CO)6 and Mn2(CO)10 in concentrated solutions of KOH/MeOH produced two new mixed Cr-Mn-carbonyl clusters, [E2CrMn2(CO)9](2-) (E=S, 1; Se, 2). Clusters 1 and 2 were isostructural with one another and each displayed a trigonal-bipyramidal structure, with the CrMn2 triangle axially capped by two µ3-E atoms. The analogous telluride cluster, [Te2CrMn2(CO)9](2-) (3), was obtained from the ring-closure of Te2Mn2 ring complex [Te2Mn2Cr2(CO)18](2-) (4). Upon bubbling with CO, clusters 2 and 3 were readily converted into square-pyramidal clusters, [E2CrMn2(CO)10](2-) (E=Se, 5; Te, 6), accompanied with the cleavage of one Cr-Mn bond. According to SQUID analysis, cluster 6 was paramagnetic, with S=1 at room temperature; however, the Se analogue (5) was spectroscopically proposed to be diamagnetic, as verified by TD-DFT calculations. Cluster 6 could be further carbonylated, with cleavage of the Mn-Mn bond to produce a new arachno-cluster, [Te2CrMn2(CO)11](2-) (7). The formation and structural isomers, as well as electrochemistry and UV/Vis absorption, of these clusters were also elucidated by DFT calculations.

Improved execution efficiency of model-based roentgen stereophotogrammetric analysis: simplification and segmentation of model meshes.

Recently, the model-based roentgen stereophotogrammetric analysis (RSA) method has been developed as an in vivo tool to estimate static pose and dynamic motion of the instrumented prostheses. The two essential inputs for the RSA method are prosthetic models and roentgen images. During RSA calculation, the implants are often reversely scanned and input in the form of meshes to estimate the outline error between prosthetic projection and roentgen images. However, the execution efficiency of the RSA iterative calculation may limit its clinical practicability, and one reason for inefficiency may be very large number of meshes in the model. This study uses two methods of mesh manipulation to improve the execution efficiency of RSA calculation. The first is to simplify the model meshes and the other is to segment and delete the meshes of insignificant regions. An index (i.e. critical percentage) of an optimal element number is defined as the trade-off between execution efficiency and result accuracy. The predicted results are numerically validated by total knee prosthetic system. The outcome shows that the optimal strategy of the mesh manipulation is simplification and followed by segmentation. On average, the element number can even be reduced to 1% of the original models. After the mesh manipulation, the execution efficiency can be increased about 75% without compromising the accuracy of the predicted RSA results (the increment of rotation and translation error: 0.06° and 0.02 mm). In conclusion, prosthetic models should be manipulated by simplification and segmentation methods prior to the RSA calculation to increase the execution efficiency and then to improve clinical applicability of the RSA method.

The expression levels of outer membrane proteins STM1530 and OmpD, which are influenced by the CpxAR and BaeSR two-component systems, play important roles in the ceftriaxone resistance of Salmonella enterica serovar Typhimurium.

Significant increases in STM3031, STM1530, and AcrD protein levels and significant decreases in OmpC and OmpD protein levels are present when the ceftriaxone-resistant Salmonella enterica serovar Typhimurium R200 strain is compared with the ceftriaxone-susceptible strain 01-4. AcrD is known to be involved in drug export, and STM3031 seems to play a key role in ceftriaxone resistance. Here, we examine the roles of STM1530, OmpC, and OmpD in ceftriaxone resistance. An ompD gene deletion mutant showed 4-fold higher ceftriaxone resistance than 01-4. An ompC gene deletion mutant showed 4-fold higher cephalothin and erythromycin resistance than 01-4, but there was no effect on ceftriaxone resistance. However, a stm1530 deletion mutant did show >64-fold lower ceftriaxone resistance than R200. Moreover, the STM3031 protein was significantly decreased in R200(Δstm1530) compared to R200. STM3031 expression has been shown to be influenced by the two-component system regulator gene baeR. CpxR seems to modulate BaeR. A cpxA-cpxR gene deletion mutant showed >2,048-fold lower ceftriaxone resistance than R200. The outer membrane protein profile of R200(ΔcpxAR) showed significant decreases in STM3031 and STM1530 compared to R200, while OmpD had returned to the level found in 01-4. Furthermore, the stm3031, stm1530, and ompD mRNA levels were correlated with their protein expression levels in these strains, while decreases in the mRNA levels of the efflux pump acrB, acrD, and acrF genes were found in R200(ΔcpxAR). Findings similar to those for R200(ΔcpxAR) were found for R200(ΔbaeSR). These results, together with those for STM3031 and the fact that STM1530 is an outer membrane protein, suggest that STM1530 and OmpD are influenced by the CpxAR and BaeSR two-component systems and that this contributes to S. enterica serovar Typhimurium ceftriaxone resistance.

Performance assessment of a nested-PCR assay (the RAPID BAP-MTB) and the BD ProbeTec ET system for detection of Mycobacterium tuberculosis in clinical specimens.

The performance of a nested PCR-based assay (the RAPID BAP-MTB; AsiaGen, Taichung, Taiwan) and the BD ProbeTec ET (DTB) system (Becton Dickinson, Sparks, Md.) for detection of Mycobacterium tuberculosis was evaluated with 600 consecutive clinical samples. These samples, including 552 respiratory specimens and 48 nonrespiratory specimens, were collected from 333 patients treated at National Taiwan University Hospital from September to October 2003. The results of both assays were compared to the gold standard of combined culture results and clinical diagnosis. The overall sensitivity and specificity of the RAPID BAP-MTB assay for respiratory specimens were 66.7% and 97.2%, respectively, and for the DTB assay they were 56.7% and 95.3%, respectively. The positive and negative predictive values for the RAPID BAP-MTB were 74.1% and 96.0%, respectively, and for the DTB assay they were 59.6% and 94.7%, respectively. For smear-negative samples, the sensitivity of the RAPID BAP-MTB and DTB assays was 57.1% and 40.5%, respectively. The RAPID BAP-MTB assay produced 14 false-positive results in 14 samples, including one of the six samples yielding Mycobacterium abscessus, one of the six samples yielding Mycobacterium avium intracellulare, one sample from a patient with a history of pulmonary tuberculosis with complete treatment, and three samples from three patients with a previous diagnosis of tuberculosis who were under treatment at the time of specimen collection. Among the 48 nonrespiratory specimens, the RAPID BAP-MTB assay was positive in one biopsy sample from a patient with lumbar tuberculous spondylitis and one pus sample from a patient with tuberculous cervical lymphadenopathy. Our results showed that the RAPID BAP-MTB assay is better than the DTB assay for both respiratory specimens and nonrespiratory specimens. The overall time for processing this assay is only 5 h. In addition, its diagnostic accuracy in smear-negative samples is as high as in smear-positive samples.