Nutritional intervention is effective in reducing the risk of neural tube defects (NTDs). To determine the effects of nutritional supplementation on human metabolism, a metabonomic study was carried out on 96 women of reproductive age. Subjects with nutritional intervention were given fortified wheat flour (containing folic acid, vitamin B(1), vitamin B(2), ferric sodium edetate and zinc oxide) for 8 months. Serum metabolic fingerprinting was detected via ultraperformance liquid chromatography in tandem with time-of-flight mass spectrometry (UPLC-TOF MS), and data acquired was processed by multivariate statistical analysis. The result revealed a significant difference between the control and intervention group. Twenty potential biomarkers, including fructose 6-phosphate, sphingosine 1-phosphate, docosahexaenoic acid and hexadecanoic acid, were located and identified by the accurate mass measurement of TOF MS. These compounds are believed to be functionally related to anti-oxidative competence in vivo. In conclusion, metabonomics study is a valuable approach in exploring the effect mechanism of nutritional intervention on NTD prevention. Copyright (c) 2010 John Wiley & Sons, Ltd.

In this study, two WRKY genes were isolated from Erysiphe necator-resistant Chinese wild Vitis pseudoreticulata W. T. Wang 'Baihe-35-1', and designated as VpWRKY1 (GenBank accession no. GQ884198) and VpWRKY2 (GenBank accession no. GU565706). Nuclear localization of the two proteins was demonstrated in onion epidermal cells, while trans-activation function was confirmed in the leaves of 'Baihe-35-1'. Expression of VpWRKY1 and VpWRKY2 was induced rapidly by salicylic acid treatment in 'Baihe-35-1'. Expression of VpWRKY1 and VpWRKY2 was also induced rapidly by E. necator infection in 11 grapevine genotypes; the maximum induction of VpWRKY1 was greater in E. necator-resistant grapevine genotypes than in susceptible ones post E. necator inoculation. Furthermore, ectopic expression of VpWRKY1 or VpWRKY2 in Arabidopsis enhanced resistance to powdery mildew Erysiphe cichoracearum, and enhanced salt tolerance of transgenic plants. VpWRKY2 also enhanced cold tolerance of transgenic plants. In addition, the two proteins were shown to regulate the expression of some defense marker genes in Arabidopsis and grapevine. The data suggest that VpWRKY1 and VpWRKY2 may underlie the resistance in transgenic grapevine to E. necator and tolerance to salt and cold stresses.

The microbial production of 1,3-propaneidol (1,3-PD) by Klebsiella pneumoniae in continuous fermentation was investigated under low, medium and high glycerol concentrations in the absence and presence of oxygen. The production of 1,3-PD increased with increasing glycerol concentrations, reaching a maximum (266 mmol l(-1)) under high glycerol concentration (760 mmol l(-1)) with air sparging at 0.04 vvm. The yield of 1,3-PD, however, decreased gradually with increasing glycerol concentrations, with the highest yield (0.52 mol mol(-1)) obtained for low glycerol concentration (270 mmol l(-1)) under anaerobic condition. Enzyme activity assays showed that the specific activity of glycerol dehydratase was highest (0.04 U mg(-1)) for culture sparged with 0.04 vvm air under high glycerol concentration. The specific activities of glycerol dehydrogenase and 1,3-propanediol oxidoreductase were also improved for all glycerol concentrations and in the presence of oxygen, implying that the dha operon was not repressed under microaerobic conditions. Analysis of metabolic fluxes showed that more carbon flux was shifted to the oxidative pathway with increasing glycerol concentrations, resulting in a reduced flux to 1,3-PD formation. However, the increases in carbon fluxes were not evenly distributed among the oxidative branches of the pathway. Furthermore, ethanol and acetic acid levels were slightly increased whereas 2,3-butanediol and lactic levels were greatly enhanced.

It is well documented that Provirus integration site for Moloney murine leukemia virus 1 (Pim-1), as a proto-oncogene encoding a serine/threonine kinase with multiple cellular functions, is tightly associated with the occurrence and development of tumors. Overexpression of Pim-1 plays a critical role in the progression of several different tumors. In this study, esophageal squamous cell carcinoma (ESCC) EC9706 cells with Pim-1 siRNA treatment resulted in a clear decrease of Pim-1 levels, followed by inhibiting cell proliferation and inducing apoptosis. Further, Pim-1 siRNA reduced phosphorylation of Akt and Bad, and increased cleaved caspase-3/-9 activities and expression levels. These data suggest that Pim-1 siRNA-mediated apoptosis is closely related to the decrease in Akt and Bad phosphorylation and increase in cleaved caspase-3/-9 activities, and thus manipulation of Pim-1 is a potential target for molecular therapy in the clinical treatment of patients with ESCC.

Telomere integrity (including telomere length and capping) is critical in overall genomic stability. Telomere repeat binding factors and their associated proteins play vital roles in telomere length regulation and end protection. In this study, we explore the protein network surrounding telomere repeat binding factors, TRF1, TRF2, and POT1 using dual-tag affinity purification in combination with multidimensional protein identification technology liquid chromatography - tandem mass spectrometry (MudPIT LC-MS/MS). After control subtraction and data filtering, we found that TRF2 and POT1 co-purified all six members of the telomere protein complex, while TRF1 identified five of six components at frequencies that lend evidence towards the currently accepted telomere architecture. Many of the known TRF1 or TRF2 interacting proteins were also identified. Moreover, putative associating partners identified for each of the three core components fell into functional categories such as DNA damage repair, ubiquitination, chromosome cohesion, chromatin modification/remodeling, DNA replication, cell cycle and transcription regulation, nucleotide metabolism, RNA processing, and nuclear transport. These putative protein-protein associations may participate in different biological processes at telomeres or, intriguingly, outside telomeres.

Current evidence suggests that angiotensin-converting enzyme inhibitors (ACEIs) and angiotensin II receptor blockers (ARBs) have emerged as novel drugs for preventing the development of atrial fibrillation (AF). A meta-analysis was performed of 26 randomized controlled clinical trials evaluating the effect of ACEIs or ARBs on the prevention of AF. Overall, ACEIs and ARBs revealed statistically significant preventive effects on AF (odds ratio (OR), 0.65; 95% confidence interval (CI), 0.55-0.76). The preventive effect was similar in the two classes of drugs (ACEI: OR, 0.68; ARB: OR, 0.69). ACEIs and ARBs showed greater preventive effects on recurrent AF (OR, 0.45; 95% CI, 0.31-0.65) than on new-onset AF (OR, 0.80; 95% CI, 0.70-0.92). Prevention was greatest in patients with AF who were receiving amiodarone as a basic treatment drug (OR, 0.35; 95% CI, 0.26-0.48). In patients with heart failure, there appeared to be a large effect (OR, 0.497), but the credible interval (CrI) limits were wide (95% CrI, 0.187-1.161).

Brain injury after focal cerebral ischemia, the most common cause of stroke, develops from a series of pathological processes, including excitotoxicity, inflammation, and apoptosis. While NMDA receptors have been implicated in excitotoxicity, attempts to prevent ischemic brain damage by blocking NMDA receptors have been disappointing. Disruption of neuroprotective pathways may be another avenue responsible for ischemic damage, and thus preservation of neuronal survival may be important for prevention of ischemic brain injury. Here, we report that suppression of proteolytic degradation of transient receptor potential canonical 6 (TRPC6) prevented ischemic neuronal cell death in a rat model of stroke. The TRPC6 protein level in neurons was greatly reduced in ischemia via NMDA receptor-dependent calpain proteolysis of the N-terminal domain of TRPC6 at Lys16. This downregulation was specific for TRPC6 and preceded neuronal death. In a rat model of ischemia, activating TRPC6 prevented neuronal death, while blocking TRPC6 increased sensitivity to ischemia. A fusion peptide derived from the calpain cleavage site in TRPC6 inhibited degradation of TRPC6, reduced infarct size, and improved behavioral performance measures via the cAMP response element-binding protein (CREB) signaling pathway. Thus, TRPC6 proteolysis contributed to ischemic neuronal cell death, and suppression of its degradation preserved neuronal survival and prevented ischemic brain damage.

Parasite dihydrofolate reductase (DHFR) and dihydropteroate synthase (DHPS) are known target enzymes of antifolate drugs used for the treatment and prophylaxis of persons with malaria. We sequenced the Plasmodium vivax dihydrofolate reductase (pvdhfr) and dihydropteroate synthase (pvdhps) genes to examine the prevalence and extent of point mutations in isolates from malaria-endemic countries. Double mutations (S58R and S117N) or quadruple mutations (F57L/I, S58R, T61M, and S117T) in the pvdhfr gene were found in isolates from Thailand (96.4%) and Myanmar (71.4%), but in only one isolate (1.0%) from Korea, where sulfadoxine-pyrimethamine has never been used. The pvdhfr point mutations correlated strongly with the pvdhps point mutations and ranged from single to triple mutations (S382A, A383G, and A553G), among isolates from Thailand, Myanmar, and Korea. These findings suggests that the prevalence of mutations in pvdhfr and pvdhps in P. vivax isolates from different malaria-endemic countries is associated with selection pressure imposed by sulfadoxine-pyrimethamine.

Activins stimulate FSH synthesis and secretion by pituitary gonadotrope cells. Activin A induction of porcine and murine FSHbeta (Fshb) gene transcription in immortalized gonadotropes is dependent on homolog of Drosophila mothers against decapentaplegic (SMAD) proteins as well as the forkhead transcription factor L2 (FOXL2). Using both heterologous and homologous cell models, we demonstrate that FOXL2 functionally synergizes with SMAD3/4 to stimulate porcine Fshb promoter-reporter activity. We further show that endogenous FOXL2 and SMAD2/3 physically interact in homologous cells. We identify two composite cis-elements of adjacent FOXL2 and SMAD binding sites in the proximal porcine Fshb promoter that mediate activin A, FOXL2, and SMAD3 actions. FOXL2 can bind these elements independently of SMADs, whereas SMAD3/4 binding requires high-affinity FOXL2 binding. Conversely, FOXL2 alone is insufficient to regulate Fshb transcription and requires SMADs to induce promoter activity. Collectively, our data suggest a model in which activins stimulate formation and nuclear accumulation of SMAD3/4 complexes, which interact with FOXL2 bound to at least two proximal promoter elements. This association stabilizes SMAD3/4 binding to adjacent SMAD binding elements. SMAD-FOXL2 complexes then mediate activation of transcription through a currently unknown mechanism. Conservation of one of the two composite cis-elements suggests that this may form part of a general mechanism whereby activins regulate Fshb subunit transcription and FSH synthesis.

The arctiin and arctigenin in the fruit of Arctium lappa L. were extracted by matrix solid-phase dispersion (MSPD) and determined by high-performance liquid chromatography (HPLC) with fluorescence detection. The experimental conditions for the MSPD were optimized. Silica gel was selected as dispersion adsorbent and methanol as elution solvent. The calibration curve showed good relationship (r>0.9998) in the concentration range of 0.010-5.0mugmL(-1) for arctiin and 0.025-7.5mugmL(-1) for arctigenin. The recoveries were between 74.4% and 100%. The proposed method consumed less sample, time and solvent compared with conventional methods, including ultrasonic and Soxhlet extraction.

PURPOSE:: To evaluate the effect of transcatheter administration of 10-hydroxycamptothecin (HCPT), a hypoxia-inducible factor-1alpha (HIF-1alpha) inhibitor, on HIF-1alpha expression and angiogenesis in liver tumors after transcatheter arterial embolization in an animal model. MATERIALS AND METHODS:: VX2 tumors were implanted in the livers of 30 rabbits. The animals were divided randomly into three groups of 10 animals each. Group 1 animals received hepatic intraarterial infusion of distilled water. Group 2 animals received iodized oil infusion followed by embolization with 150-250 mum of polyvinyl alcohol particles. Group 3 animals received infusion of a mixture of HCPT (1 mg/kg body weight) with iodized oil followed by the particle embolization. Six hours or 3 days after transcatheter treatment, the animals were sacrificed, and the tumor samples were harvested. Immunohistochemical staining was performed to evaluate the levels of HIF-1alpha and vascular endothelial growth factor (VEGF) protein as well as microvessel density. RESULTS:: The levels of HIF-1alpha and VEGF and microvessel density in tumors of group 2 were significantly higher than those of group 1 or 3 (P < .05). However, no significant differences were noted in tumors between group 1 and 3 (P > .05). HIF-1alpha levels were significantly correlated with VEGF levels (r = .587, P = .001) and microvessel density (r = .527, P = .003). CONCLUSIONS:: Transcatheter infusion of HCPT has an inhibitory effect on HIF-1alpha expression and angiogenesis in liver tumors after transcatheter arterial embolization.

Co(3)O(4) nanofibers were fabricated by a two-step procedure consisting of electrospinning and subsequent calcination. Scanning electron microscopy and transmission electron microscopy were employed to characterize the as-prepared Co(3)O(4) nanofibers. Fourier transform infrared spectroscopy and Raman spectroscopy were used to confirm the degradation of the polymer matrix and the complete conversion of cobalt nitrate to cobalt oxide. Furthermore, the composition and crystal structure of the final product were investigated using X-ray diffraction and X-ray photoelectron spectroscopy. The as-prepared Co(3)O(4) nanofibers were applied to construct a non-enzymatic sensor for glucose detection in alkaline solution. The developed sensor showed a fast response time (less than 7s), a high sensitivity of 36.25muAmM(-1)cm(-2), good reproducibility and selectivity, and a detection limit of 0.97muM (S/N=3). The high concentration of NaCl does not poison the electrode. Its application for the detection of glucose in human blood serum sample shows good agreement with the results obtained from commercial glucose meter. The Langmuir isothermal theory was employed to fit the obtained calibration curve. The mechanisms for the glucose oxidation promoted by Co(3)O(4) nanofibers and the good selectivity against uric acid and ascorbic acid at an applied potential of +0.59V vs. Ag/AgCl were also proposed. These results demonstrate that Co(3)O(4) nanofibers have great potential applications in the development of sensors for enzyme-free detection of glucose.

Phytochemical investigation of the methanolic extract from the aerial parts of Agrimonia pilosa led to the isolation of three compounds, (-)-aromadendrin 3-O-beta-d-glucopyranoside (1), desmethylagrimonolide 6-O-beta-d-glucopyranoside (2), and 5,7-dihydroxy-2-propylchromone 7-O-beta-d-glucopyranoside (3), together with nine known compounds, agrimonolide 6-O-glucoside, takanechromone C, astragalin, afzelin, tiliroside, luteolin, quercetin, isoquercetrin, and quercitrin. Their structures were determined by various spectroscopic analysis and chemical transformations.

OBJECTIVE: A series of research works had proven that high-resolution magnetic resonance imaging (MRI) was an effective tool to identify the morphological appearance of atherosclerotic vulnerable plaque. We performed further study to compare ex vivo vascular morphology with MRI and histological examination using the specimens of carotid endarterectomy and femoral artery (FA) prosthetic reconstruction, and described the detailed histology and immunohistochemistry features ofplaque. METHODS: Eight consecutive patients (five males, mean age 67.4 years) who were recruited into our study had high-grade stenosis or occlusion of carotid artery and FA (70 to 100%). Four patients accepted carotid endarterectomy and others underwent FA prosthetic reconstruction. RESULTS: The high level of accuracy was shown in classification of the plaque type. The sections of plaques detected by MRI were classified as follows: 12 (22.2%) were modified American Heart Association (AHA) type III, 11 (20.4%) were type IV-V, 26 (48.2%) were type VI and 5 (9.3%) were type VII. According to the histological observations, the overall accurate rate by MRI showed a high level accuracy of 90.7% (49/54). Immunohistochemistry of cellular components in plaque shoulder demonstrated that CD68-positive macrophages were strongly dominated along the arterial wall. Reduced expression of smooth muscle actin (SMA)-positive smooth muscle cells were detected in the fibrous cap of atheroma. CD31 stain showed the increased presence of CD31-positive neomicrovessels. CONCLUSIONS: Atherosclerosis is a systematic disease that can involve whole body vascular beds. Noninvasive high-resolution MRI will provide useful information in morphological studies of the arterial walls and classification of the plaque type.

Various in situ injectable hydrogels have been developed for protein delivery in treating human diseases. However, most hydrogels are highly permeable, which can lead to the rapid release of loaded proteins. The purpose of this study is to apply nucleic acid aptamers to functionalize an in situ injectable hydrogel model to control the release of proteins. The aptamers were studied using secondary structural predictions and binding analyses. The results showed that the structural predictions were different from the experimental measurements in numerous cases. The affinity of the aptamer was significantly affected by the mutations of the essential nucleotides, whereas it was not significantly affected by the variations of the nonessential nucleotides. The mutated aptamers were then used to functionalize the injectable hydrogel model. The results showed that the aptamer-functionalized hydrogel could prolong protein release. Moreover, the release rates could be controlled by adjusting the affinity of the aptamer.

BACKGROUND: Sorafenib and sunitinib are novel small molecule tyrosine kinase inhibitors with multiple targets on tumor cells, which have been demonstrated to be beneficial in the treatment of several carcinomas. Combining the usage of molecular targeted agents and adoptive cellular immunotherapy (ACI) against drug-resistant relapse nasopharyngeal carcinoma which had no standard therapeutic regimen was investigated by our research in order to study whether synergistic effects exist and related mechanisms. METHODS: Human multidrug-resistant nasopharyngeal carcinoma cell line CNE2/DDP with high and low expressions of ABCG(2) (abbreviated to ABCG (2) (High) CNE2/DDP and ABCG (2) (Low) CNE2/DDP) cells and NK cells were isolated by magnetic activated cell sorting, and the purity of isolated cells was detected by flow cytometry. mRNA expressions of drug-resistant gene ABCG(2), Bcl-2, MDR1, MRP and MGMT in ABCG (2) (High) CNE2/DDP and ABCG (2) (Low) CNE2/DDP cells were detected by reverse transcription polymerase chain reaction (RT-PCR). Drug sensitivity of two kinds of cells to fluorouracil, cisplatin, vincristine, carboplatin, epirubicin, daunorubicin, paclitaxel, mitomycin, sorafenib, and sunitinib were detected by MTT assay. FCM was used to evaluate the expressions of NKG2D ligands (NKG2DLs,) on target cells before and after incubated with sorafenib and sunitinib. Subsequently, the cytotoxic sensitivity of incubated and un-incubated ABCG (2) (High) CNE2/DDP and ABCG (2) (Low) CNE2/DDP cells to NK cells was measured by CytoTox 96((R)) Non-Radioactive Cytotoxicity Assay. RESULTS: The results revealed that target cells' cytotoxic sensitivity to natural killer (NK) cells increased in association with up-regulation of NKG2DLs on tumor cells after incubation with sorafenib and sunitinib. Furthermore, up-regulation in sunitinib group was much higher than in sorafenib group when it came to the expressions of NKG2DLs on tumor cells. For another, ABCG (2) (High) CNE2/DDP was much more sensitive to the regulation than ABCG (2) (Low) CNE2/DDP. CONCLUSIONS: Our research revealed for the first time that sorafenib and sunitinib could up-regulate NKG2DLs on tumor cells resulting in markedly increased tumor cells cytotoxic sensitivity to NK cells, which suggested that combining usage of molecular targeted agents and ACI may result in great benefits in clinical practice for the therapy-resistant cases and drug-resistant relapse.

Structured light illumination by means of phase shifting patterns is a widely employed method for three-dimensional (3-D) image acquisition that is robust to ambient light and object albedo but may be especially susceptible to sensor and environment noise. In this paper, we study the specific technique of phase measuring profilometry (PMP) and the maximization of a pattern's signal to noise ratio (SNR). By treating the design of an N-pattern PMP process as placing points in an N-dimensional coding space, we define a pattern's SNR in terms of a pattern set's computational length and the number of coded phase periods in the projected patterns. Then, without introducing phase ambiguities, we propose a so-called edge pattern strategy that maximizes the computational length and number of periods. Theoretically, the edge pattern technique improves the SNR by 1.2381 times when using three component patterns and by 15.5421 times when using five patterns. Experimental results further demonstrate the improved SNR of the proposed edge pattern technique such that more accurate 3-D results are achieved using fewer component patterns.

We derive the fourth-order oriented partial-differential equations (PDEs) for noise removal of two-photon fluorescence images. We consider it from two aspects: one is based on a variational method; the other is based on controlling the diffusion direction. Our filtering model makes the diffusion along only the special orientation-decided via all the information in the established filtering window, so the edges are protected during filtering. Compared with related PDEs models, our model shows superior performance in terms of both objective criteria and subjective human vision via processing simulated and experimental noisy images.

The study of Lamb shift plays a unique role in quantum electrodynamics because it provides an excellent test of the theory on photonic crystals (PCs). In this Letter, we present the first observation of large Lamb shift in the YBO(3):Eu(3+) inverse opals fabricated by the polystyrene templating method. In addition, it is very interesting to observe that the luminescent dynamics of Eu(3+) decayed with a faster power law (t(-3)), followed by a slower exponential process due to the coexistence of the diffusion field and the propagating field in the PCs.

KRAS mutations are found in approximately 90% of human pancreatic ductal adenocarcinomas (PDAC). However, mice genetically engineered to express Kras(G12D) from its endogenous locus develop PDACs only after a prolonged latency, indicating that other genetic events or pathway alterations are necessary for PDAC progression. The PTEN-controlled phosphatidylinositol 3-kinase (PI3K)/AKT signaling axis is dysregulated in later stages of PDAC. To better elucidate the role of PTEN/PI3K/AKT signaling in Kras(G12D)-induced PDAC development, we crossed Pten conditional knockout mice (Pten(lox/lox)) to mice with conditional activation of Kras(G12D). The resulting compound heterozygous mutant mice showed significantly accelerated development of acinar-to-ductal metaplasia (ADM), malignant pancreatic intraepithelial neoplasia (mPanIN), and PDAC within a year. Moreover, all mice with Kras(G12D) activation and Pten homozygous deletion succumbed to cancer by 3 weeks of age. Our data support a dosage-dependent role for PTEN, and the resulting dysregulation of the PI3K/AKT signaling axis, in both PDAC initiation and progression, and shed additional light on the signaling mechanisms that lead to the development of ADM and subsequent mPanIN and pancreatic cancer. Cancer Res; 70(18); OF1-11. (c)2010 AACR.