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Find video protocols related to scientific articles indexed in Pubmed.
Silencing of OPR3 in tomato reveals the role of OPDA in callose deposition during activation of defense responses against Botrytis cinerea.
Plant J.
PUBLISHED: 11-06-2014
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OPDA likely plays signaling roles in plant defense that do not depend on its further conversion to the phytohormone jasmonic acid. To elucidate the role of OPDA in tomato plant defense, we have silenced the 12-oxophytodienoate reductase 3 (OPR3) gene. Two independent transgenic tomato lines (SiOPR3-1 and SiOPR3-2) showed significantly reduced OPR3 expression upon infection with the necrotrophic pathogen Botrytis cinerea. Moreover, SiOPR3 plants are more susceptible to this pathogen, and this susceptibility is accompanied by a significant decrease in the OPDA levels and an almost abolished JA-Ile production. OPR3 silencing also leads to a major reduction in the expression of other genes of the jasmonic acid (JA) synthesis and signaling pathways after infection. These results confirm that in tomato plants, as in Arabidopsis, OPR3 determines OPDA availability for JA biosynthesis. In addition, we show that an intact JA biosynthetic pathway is required for proper callose deposition since its pathogen-induced accumulation is reduced in SiOPR3 plants. Interestingly, OPDA, but not JA, treatment restored basal resistance to B. cinerea and induced callose deposition in SiOPR3-1 and SiOPR3-2 transgenic plants. These results provide clear evidence that OPDA by itself plays a major role in basal defense of tomato plants against this necrotrophic pathogen. This article is protected by copyright. All rights reserved.
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Absence of endo-1,4-?-glucanase KOR1 alters the jasmonate-dependent defence response to Pseudomonas syringae in Arabidopsis.
J. Plant Physiol.
PUBLISHED: 07-23-2014
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During plant-pathogen interactions, the plant cell wall forms part of active defence against invaders. In recent years, cell wall-editing enzymes, associated with growth and development, have been related to plant susceptibility or resistance. Our previous work identified a role for several tomato and Arabidopsis endo-1,4-?-glucanases (EGs) in plant-pathogen interactions. Here we studied the response of the Arabidopsis thaliana T-DNA insertion mutant lacking EG Korrigan1 (KOR1) infected with Pseudomonas syringae. KOR1 is predicted to be an EG which is thought to participate in cellulose biosynthesis. We found that kor1-1 plants were more susceptible to P. syringae, and displayed severe disease symptoms and enhanced bacterial growth if compared to Wassilewskija (Ws) wild-type plants. Hormonal and gene expression analyses revealed that the jasmonic acid (JA) pathway was activated more in kor1-1 plants with an increase in the JA-biosynthesis gene LOX3 and a greater accumulation of JA. Upon infection the accumulation of JA and JA-isoleucine (JA-Ile) was higher than in wild-type plants and increased the induction of LOX3 and the JA-responsive PDF1.2 gene. In addition, the increase of salicylic acid (SA) in healthy and infected kor1-1 may reflect the complex interaction between JA and SA, which results in the more susceptible phenotype displayed by the infected mutant plants. Callose deposition was enhanced in infected kor1-1 and an increase in pathogen-induced hydrogen peroxide took place. The susceptible phenotype displayed by KOR1-deficient plants was coronatine-independent. No significant changes were detected in the hormonal profile of the kor1-1 plants infected by coronatine-deficient P. syringae cmaA, which supports that absence of EG KOR1 alters per se the plant response to infection. We previously reported increased resistance of kor1-1 to B. cinerea, hence, the lack of this EG alters cell wall properties and plant responses in such a way that benefits P. syringae colonisation but restricts B. cinerea invasion.
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The deubiquitinating enzyme USP5 modulates neuropathic and inflammatory pain by enhancing Cav3.2 channel activity.
Neuron
PUBLISHED: 07-10-2014
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T-type calcium channels are essential contributors to the transmission of nociceptive signals in the primary afferent pain pathway. Here, we show that T-type calcium channels are ubiquitinated by WWP1, a plasma-membrane-associated ubiquitin ligase that binds to the intracellular domain III-IV linker region of the Cav3.2 T-type channel and modifies specific lysine residues in this region. A proteomic screen identified the deubiquitinating enzyme USP5 as a Cav3.2 III-IV linker interacting partner. Knockdown of USP5 via shRNA increases Cav3.2 ubiquitination, decreases Cav3.2 protein levels, and reduces Cav3.2 whole-cell currents. In vivo knockdown of USP5 or uncoupling USP5 from native Cav3.2 channels via intrathecal delivery of Tat peptides mediates analgesia in both inflammatory and neuropathic mouse models of mechanical hypersensitivity. Altogether, our experiments reveal a cell signaling pathway that regulates T-type channel activity and their role in nociceptive signaling.
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Electrochemical immunosensor for anti-tissue transglutaminase antibodies based on the in situ detection of quantum dots.
Talanta
PUBLISHED: 07-10-2014
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A miniaturized electrochemical biosensor array with in situ detection of quantum dots (QDs) was developed for the detection of anti-transglutaminase IgG antibodies (a celiac disease biomarker) in human sera. For the fabrication of the sensor, a 8-channel screen-printed carbon electrochemical arrays were used as transducers and modified with tissue-transglutaminase by adsorption. The immunologic reaction was carried out in a few simple steps: reaction with human serum, which contains the analyte of interest, followed by the immunoreaction with anti-human IgG labeled with CdSe/ZnS QDs and electrochemical detection of Cd(2+) released from QDs. All steps were performed on the screen-printed arrays as the solid support, and the detection of Cd(2+) was performed in situ after acid attack of the QDs without a transfer step by voltammetric stripping. The electrochemical response was correlated with the anti-transglutaminase IgG concentration. The developed electrochemical immunosensor is a trustful screening tool for celiac disease diagnosis discriminating between positive and negative sera samples with high sensitivity.
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Angiogenesis and proliferation markers in adjacent cirrhotic tissue could predict hepatocellular carcinoma outcome after liver transplantation.
Eur J Gastroenterol Hepatol
PUBLISHED: 06-13-2014
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The current staging systems for hepatocellular carcinoma (HCC) do not sufficiently predict outcomes after liver transplantation (LT). The present study assessed whether some tissue markers related to proliferation and angiogenesis have prognostic value.
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ATP-mediated kinome selectivity: the missing link in understanding the contribution of individual JAK Kinase isoforms to cellular signaling.
ACS Chem. Biol.
PUBLISHED: 05-28-2014
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Kinases constitute an important class of therapeutic targets being explored both by academia and the pharmaceutical industry. The major focus of this effort was directed toward the identification of ATP competitive inhibitors. Although it has long been recognized that the intracellular concentration of ATP is very different from the concentrations utilized in biochemical enzyme assays, little thought has been devoted to incorporating this discrepancy into our understanding of translation from enzyme inhibition to cellular function. Significant work has been dedicated to the discovery of JAK kinase inhibitors; however, a disconnect between enzyme and cellular function is prominently displayed in the literature for this class of inhibitors. Herein, we demonstrate utilizing the four JAK family members that the difference in the ATP Km of each individual kinase has a significant impact on the enzyme to cell inhibition translation. We evaluated a large number of JAK inhibitors in enzymatic assays utilizing either 1 mM ATP or Km ATP for the four isoforms as well as in primary cell assays. This data set provided the opportunity to examine individual kinase contributions to the heterodimeric kinase complexes mediating cellular signaling. In contrast to a recent study, we demonstrate that for IL-15 cytokine signaling it is sufficient to inhibit either JAK1 or JAK3 to fully inhibit downstream STAT5 phosphorylation. This additional data thus provides a critical piece of information explaining why JAK1 has incorrectly been thought to have a dominant role over JAK3. Beyond enabling a deeper understanding of JAK signaling, conducting similar analyses for other kinases by taking into account potency at high ATP rather than Km ATP may provide crucial insights into a compound's activity and selectivity in cellular contexts.
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Identification of (R)-6-(1-(4-cyano-3-methylphenyl)-5-cyclopentyl-4,5-dihydro-1H-pyrazol-3-yl)-2-methoxynicotinic acid, a highly potent and selective nonsteroidal mineralocorticoid receptor antagonist.
J. Med. Chem.
PUBLISHED: 04-30-2014
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A novel series of nonsteroidal mineralocorticoid receptor (MR) antagonists identified as part of our strategy to follow up on the clinical candidate PF-03882845 (2) is reported. Optimization departed from the previously described pyrazoline 3a and focused on improving the selectivity for MR versus the progesterone receptor (PR) as an approach to avoid potential sex-hormone-related adverse effects and improving biopharmaceutical properties. From this effort, (R)-14c was identified as a potent nonsteroidal MR antagonist (IC50 = 4.5 nM) with higher than 500-fold selectivity versus PR and other related nuclear hormone receptors, with improved solubility as compared to 2 and pharmacokinetic properties suitable for oral administration. (R)-14c was evaluated in vivo using the increase of urinary Na(+)/K(+) ratio in rat as a mechanism biomarker of MR antagonism. Treatment with (R)-14c by oral administration resulted in significant increases in urinary Na(+)/K(+) ratio and demonstrated this novel compound acts as an MR antagonist.
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Electrochemical biotin detection based on magnetic beads and a new magnetic flow cell for screen printed electrode.
Talanta
PUBLISHED: 04-25-2014
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The use of the first flow-cell for magnetic assays with an integrated magnet is reported here. The flow injection analysis system (FIA) is used for biotin determination. The reaction scheme is based on a one step competitive assay between free biotin and biotin labeled with horseradish peroxidase (B-HRP). The mixture of magnetic beads modified with streptavidin (Strep-MB), biotin and B-HRP is left 15 min under stirring and then a washing step is performed. After that, 100 ?L of the mixture is injected and after 30s 100 ?L of 3,3',5,5'-Tetramethylbenzidine (TMB) is injected and the FIAgram is recorded applying a potential of -0.2V. The linear range obtained is from 0.01 to 1 nM of biotin and the sensitivity is 758 nA/nM. The modification and cleaning of the electrode are performed in an easy way due to the internal magnet of the flow cell.
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Phase I clinical trial of temsirolimus and vinorelbine in advanced solid tumors.
Cancer Chemother. Pharmacol.
PUBLISHED: 03-31-2014
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To determine the maximal tolerated dose (MTD) of the combination of weekly temsirolimus and every other week vinorelbine in patients with advanced or refractory solid tumors.
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Multiple determinations of sperm DNA fragmentation show that varicocelectomy is not indicated for infertile patients with subclinical varicocele.
Biomed Res Int
PUBLISHED: 02-06-2014
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Varicocele is one of the most common causes of low semen quality, which is reflected in high percentages of sperm cells with fragmented DNA. While varicocelectomy is usually performed to ameliorate a patient's fertility, its impact on sperm DNA integrity in the case of subclinical varicocele is poorly documented. In this study, multiple DNA fragmentation analyses (TUNEL, SCD, and SCSA) were performed on semen samples from sixty infertile patients with varicocele (15 clinical varicoceles, 19 clinical varicoceles after surgical treatment, 16 subclinical varicoceles, and 10 subclinical varicoceles after surgical treatment). TUNEL, SCD, and SCSA assays all showed substantial sperm DNA fragmentation levels that were comparable between subclinical and clinical varicocele patients. Importantly, varicocelectomy did improve sperm quality in patients with clinical varicocele; however, this was not the case in patients with subclinical varicocele. In summary, although infertile patients with clinical and subclinical varicocele have similar sperm DNA quality, varicocelectomy should only be advised for patients with clinical varicocele.
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Effect of location on primary stability and healing of dental implants.
Implant Dent
PUBLISHED: 01-04-2014
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To study implant primary stability and bone healing using resonance frequency analysis in different anatomical locations 4 months after placement.
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Priming of plant resistance by natural compounds. Hexanoic acid as a model.
Front Plant Sci
PUBLISHED: 01-01-2014
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Some alternative control strategies of currently emerging plant diseases are based on the use of resistance inducers. This review highlights the recent advances made in the characterization of natural compounds that induce resistance by a priming mechanism. These include vitamins, chitosans, oligogalacturonides, volatile organic compounds, azelaic and pipecolic acid, among others. Overall, other than providing novel disease control strategies that meet environmental regulations, natural priming agents are valuable tools to help unravel the complex mechanisms underlying the induced resistance (IR) phenomenon. The data presented in this review reflect the novel contributions made from studying these natural plant inducers, with special emphasis placed on hexanoic acid (Hx), proposed herein as a model tool for this research field. Hx is a potent natural priming agent of proven efficiency in a wide range of host plants and pathogens. It can early activate broad-spectrum defenses by inducing callose deposition and the salicylic acid (SA) and jasmonic acid (JA) pathways. Later it can prime pathogen-specific responses according to the pathogen's lifestyle. Interestingly, Hx primes redox-related genes to produce an anti-oxidant protective effect, which might be critical for limiting the infection of necrotrophs. Our Hx-IR findings also strongly suggest that it is an attractive tool for the molecular characterization of the plant alarmed state, with the added advantage of it being a natural compound.
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Resistance inducers modulate Pseudomonas syringae pv. tomato strain DC3000 response in tomato plants.
PLoS ONE
PUBLISHED: 01-01-2014
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The efficacy of hexanoic acid (Hx) as an inducer of resistance in tomato plants against Pseudomonas syringae pv. tomato DC3000 was previously demonstrated, and the plant response was characterized. Because little is known about the reaction of the pathogen to this effect, the goal of the present work was to determine whether the changes in the plant defence system affect the pathogen behaviour. This work provides the first demonstration of the response of the pathogen to the changes observed in plants after Hx application in terms of not only the population size but also the transcriptional levels of genes involved in quorum sensing establishment and pathogenesis. Therefore, it is possible that Hx treatment attenuates the virulence and survival of bacteria by preventing or diminishing the appearance of symptoms and controlling the growth of the bacteria in the mesophyll. It is interesting to note that the gene transcriptional changes in the bacteria from the treated plants occur at the same time as the changes in the plants. Hx is able to alter bacteria pathogenesis and survival only when it is applied as a resistance inducer because the changes that it promotes in plants affect the bacteria.
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Comparison of microstructures between block grafts from the mandibular ramus and calvarium for horizontal bone augmentation of the maxilla: a case series study.
Int J Periodontics Restorative Dent
PUBLISHED: 10-12-2013
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The primary purpose of this clinical study was to compare architectural metric parameters using microcomputed tomography (micro-CT) between sites grafted with blocks harvested from the mandibular ramus and calvarium for horizontal bone augmentation in the maxilla. The second aim was to compare the primary stability of implants placed in both types of block grafts. Ten consecutive healthy partially edentulous patients requiring extensive horizontal bone reconstruction in the maxilla were included. A total of 14 block grafts (7 each from the mandibular ramus and calvarium) were studied. After 4 to 6 months of healing, 41 implants were placed: 24 implants (58.5%) in calvarial (group 1) and 17 (41.5%) in ramus grafts (group 2). A resonance frequency analysis (RFA) was performed to test implant stability. Furthermore, two biopsy specimens were randomly selected for histomorphometric analysis. Micro-CT analyses showed no significant difference in the morphometric parametric values analyzed between groups. Furthermore, RFA also showed no difference between groups. However, slightly higher RFA values were noted for implants placed in ramus grafts. Bone quality, as assessed by micro-CT and histomorphometric analyses, was similar in both ramus and calvarial block grafts. In addition, there was no difference in primary implant stability between groups.
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Two schedules of etirinotecan pegol (NKTR-102) in patients with previously treated metastatic breast cancer: a randomised phase 2 study.
Lancet Oncol.
PUBLISHED: 10-04-2013
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New therapeutic options are needed for patients with heavily pretreated breast cancer. Etirinotecan pegol is a long-acting topoisomerase-I inhibitor designed to provide prolonged tumour-cell exposure to SN38, the active metabolite. We aimed to assess the efficacy and safety of two etirinotecan pegol dosing schedules in patients with previously treated metastatic breast cancer to determine an optimum dosing schedule for phase 3 trials.
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Fine tuning of reactive oxygen species homeostasis regulates primed immune responses in Arabidopsis.
Mol. Plant Microbe Interact.
PUBLISHED: 10-04-2013
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Selected stimuli can prime the plant immune system for a faster and stronger defense reaction to pathogen attack. Pretreatment of Arabidopsis with the chemical agent ?-aminobutyric acid (BABA) augmented H2O2 and callose production after induction with the pathogen-associated molecular pattern (PAMP) chitosan, or inoculation with the necrotrophic fungus Plectosphaerella cucumerina. However, BABA failed to prime H2O2 and callose production after challenge with the bacterial PAMP Flg22. Analysis of Arabidopsis mutants in reactive oxygen species (ROS) production (rbohD) or ROS scavenging (pad2, vtc1, and cat2) suggested a regulatory role for ROS homeostasis in priming of chitosan- and P. cucumerina-inducible callose and ROS. Moreover, rbohD and pad2 were both impaired in BABA-induced resistance against P. cucumerina. Gene expression analysis revealed direct induction of NADPH/respiratory burst oxidase protein D (RBOHD), ?-glutamylcysteine synthetase 1 (GSH1), and vitamin C defective 1 (VTC1) genes after BABA treatment. Conversely, ascorbate peroxidase 1 (APX1) transcription was repressed by BABA after challenge with chitosan or P. cucumerina, probably to provide a more oxidized environment in the cell and facilitate augmented ROS accumulation. Measuring ratios between reduced and oxidized glutathione confirmed that augmented defense expression in primed plants is associated with a more oxidized cellular status. Together, our data indicate that an altered ROS equilibrium is required for augmented defense expression in primed plants.
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Randomized multicenter phase II trial comparing two schedules of etirinotecan pegol (NKTR-102) in women with recurrent platinum-resistant/refractory epithelial ovarian cancer.
J. Clin. Oncol.
PUBLISHED: 09-30-2013
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Etirinotecan pegol (NKTR-102) is a unique, long-acting topoisomerase-I inhibitor with prolonged systemic exposure to SN38 (7-ethyl-10-hydroxycamptothecin), the active metabolite of irinotecan. This randomized phase II trial investigated two dosing schedules of etirinotecan pegol in patients with platinum-resistant/refractory ovarian carcinoma.
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Design and synthesis of aryl sulfonamide-based nonsteroidal mineralocorticoid receptor antagonists.
Bioorg. Med. Chem. Lett.
PUBLISHED: 08-28-2013
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Hit-to-lead medicinal chemistry efforts are described starting from a screening hit 1, leading to a new class of aryl sulfonamide-based MR antagonist, exemplified by 17, that possesses favourable MR binding affinity, selectivity profile against closely related NHRs, physicochemical properties and metabolic stability.
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A systematic review on marginal bone loss around short dental implants (<10 mm) for implant-supported fixed prostheses.
Clin Oral Implants Res
PUBLISHED: 07-03-2013
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This systematic review aimed to evaluate the effect of implant length on peri-implant marginal bone loss (MBL) and its associated influencing factors.
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Combining Hexanoic Acid Plant Priming with Bacillus thuringiensis Insecticidal Activity against Colorado Potato Beetle.
Int J Mol Sci
PUBLISHED: 04-25-2013
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Interaction between insect herbivores and host plants can be modulated by endogenous and exogenous compounds present in the source of food and might be successfully exploited in Colorado potato beetle (CPB) pest management. Feeding tests with CPB larvae reared on three solanaceous plants (potato, eggplant and tomato) resulted in variable larval growth rates and differential susceptibility to Bacillus thuringiensis Cry3Aa toxin as a function of the host plant. An inverse correlation with toxicity was observed in Cry3Aa proteolytic patterns generated by CPB midgut brush-border membrane vesicles (BBMV) from Solanaceae-fed larvae, being the toxin most extensively proteolyzed on potato, followed by eggplant and tomato. We found that CPB cysteine proteases intestains may interact with Cry3Aa toxin and, in CPB BBMV from larvae fed all three Solanaceae, the toxin was able to compete for the hydrolysis of a papain substrate. In response to treatment with the JA-dependent plant inducer Hexanoic acid (Hx), we showed that eggplant reduced OPDA basal levels and both, potato and eggplant induced JA-Ile. CPB larvae feeding on Hx-induced plants exhibited enhanced Cry3Aa toxicity, which correlated with altered papain activity. Results indicated host-mediated effects on B. thuringiensis efficacy against CPB that can be enhanced in combination with Hx plant induction.
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Electrochemical determination of mercury: a review.
Talanta
PUBLISHED: 04-24-2013
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Mercury is a metal that has been extensively studied, in large part due to its high toxicity. Therefore, mercury levels must be monitored in different sample types using analytical methods. This review summarizes the electrochemical methods that have been used for mercury analysis in a variety of samples. A critical evaluation of the methods and electrode materials employed for mercury analysis is presented according to the following classifications: bare electrodes, chemically modified electrodes and nanostructured electrodes. The advantages and disadvantages of each type of electrode material regarding mercury analysis are also presented.
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Influence of Atrophic Posterior Maxilla Ridge Height on Bone Density and Microarchitecture.
Clin Implant Dent Relat Res
PUBLISHED: 04-24-2013
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PURPOSE: There is limited evidence available on the influence of residual ridge height (RH) on bone density. Therefore, this study aimed to investigate the correlation between the atrophic posterior RH in the maxilla and its bone density as determined by microcomputed tomography (?-CT). MATERIAL AND METHODS: Thirty-two subjects with atrophic posterior maxilla of residual RH <8?mm were included in this study. A preoperative cone beam CT scan with a radiographic stent was taken for each patient. A bone core biopsy was thus obtained from the predetermined surgical site. Out of 32 biopsies, 27 were intact and sent for ?-CT analysis. RESULTS: A statistically significant positive correlation between bone volumetric fraction (BV/TV) and RH was identified (r?=?0.417, p?=?.03). A statistically significant negative correlation between trabecular pattern factor and RH was also found (r?=?-0.415, p?=?.03). The rest of the morphometric parameters analyzed did not have any significant correlation to RH. CONCLUSION: BV/TV is potentially influenced by the residual bone height at the posterior maxilla. The lesser the RH, the lower the bone quantity and quality present.
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Eribulin mesylate versus ixabepilone in patients with metastatic breast cancer: a randomized Phase II study comparing the incidence of peripheral neuropathy.
Breast Cancer Res. Treat.
PUBLISHED: 04-04-2013
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Peripheral neuropathy is a common toxicity associated with tubulin-targeted chemotherapeutic agents. This Phase II study compares the incidence and severity of neuropathy associated with eribulin mesylate or ixabepilone in metastatic breast cancer (MBC). The primary objective was to assess the incidence of neuropathy; the study was designed to detect a difference in neuropathy rate of 35 % for eribulin versus 63 % for ixabepilone (odds ratio 0.316, 80 % power, 0.05 two-sided significance level). Eligibility criteria included: MBC; prior taxane therapy; at least one chemotherapy for advanced disease; no or minimal pre-existing neuropathy (Grade 0 or 1). The intent-to-treat population comprised 104 patients randomized (1:1) to eribulin mesylate (1.4 mg/m(2), 2-5 min intravenous on days 1 and 8) or ixabepilone (40 mg/m(2), 3 h intravenous on day 1) on a 21-day cycle. 101 patients in the safety population received a median of 5.0 eribulin and 3.5 ixabepilone cycles. Incidence of neuropathy (any grade) was 33.3 and 48.0 %, and peripheral neuropathy was 31.4 and 44.0 % for eribulin and ixabepilone, respectively. After controlling for pre-existing neuropathy and number of prior chemotherapies, these differences were not significant. Compared with ixabepilone, fewer patients receiving eribulin discontinued treatment due to neuropathy (3.9 vs. 18.0 %) or adverse events (AEs) in general (11.8 vs. 32.0 %). Time to onset of neuropathy was 35.9 weeks for eribulin and 11.6 weeks for ixabepilone, and time to resolution was 48 versus 10 weeks, respectively; other AEs were comparable. Objective responses were 15.4 versus 5.8 % and clinical benefit rates were 26.9 versus 19.2 %. In conclusion, after controlling for pre-existing neuropathy and number of prior chemotherapies, the differences in the incidence of neuropathy with eribulin and ixabepilone were not statistically significant. Onset of neuropathy tended to occur later with eribulin and resolve later.
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Functional analysis of endo-1,4-?-glucanases in response to Botrytis cinerea and Pseudomonas syringae reveals their involvement in plant-pathogen interactions.
Plant Biol (Stuttg)
PUBLISHED: 03-25-2013
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Plant cell wall modification is a critical component in stress responses. Endo-1,4-?-glucanases (EGs) take part in cell wall editing processes, e.g. elongation, ripening and abscission. Here we studied the infection response of Solanum lycopersicum and Arabidopsis thaliana with impaired EGs. Transgenic TomCel1 and TomCel2 tomato antisense plants challenged with Pseudomonas syringae showed higher susceptibility, callose priming and increased jasmonic acid pathway marker gene expression. These two EGs could be resistance factors and may act as negative regulators of callose deposition, probably by interfering with the defence-signalling network. A study of a set of Arabidopsis EG T-DNA insertion mutants challenged with P. syringae and Botrytis cinerea revealed that the lack of other EGs interferes with infection phenotype, callose deposition, expression of signalling pathway marker genes and hormonal balance. We conclude that a lack of EGs could alter plant response to pathogens by modifying the properties of the cell wall and/or interfering with signalling pathways, contributing to generate the appropriate signalling outcomes. Analysis of microarray data demonstrates that EGs are differentially expressed upon many different plant-pathogen challenges, hormone treatments and many abiotic stresses. We found some Arabidopsis EG mutants with increased tolerance to osmotic and salt stress. Our results show that impairing EGs can alter plant-pathogen interactions and may contribute to appropriate signalling outcomes in many different biotic and abiotic plant stress responses.
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Bevacizumab and ovarian cancer.
Ther Adv Med Oncol
PUBLISHED: 03-02-2013
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Bevacizumab is a recombinant humanized monoclonal IgG1 antibody that targets vascular endothelial growth factor-A, and is indicated in the treatment of various tumors (colon, lung, renal, and glioblastoma). It has been recently approved for the treatment of ovarian cancer in various countries. This review summarizes the activity and toxicity of bevacizumab in the treatment of ovarian cancer, both as single-agent drug and in combination with cytotoxic chemotherapy. As a single-agent drug, it has shown response rates of 16-21% in the treatment of recurrent ovarian cancer. Two phase III randomized trials have been published evaluating the addition of bevacizumab to standard chemotherapy as front-line treatment of advanced ovarian cancer. In addition, trials evaluating the combination with chemotherapy in recurrent ovarian cancer (platinum-sensitive and platinum-resistant disease) have also been reported. All these trials showed a statistically significant improvement in progression-free survival although no improvement in overall survival has been reported. The main adverse event is hypertension. Other serious, but uncommon adverse events include gastrointestinal perforation as well as renal and central nervous system toxicity.
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An electrochemical deamidated gliadin antibody immunosensor for celiac disease clinical diagnosis.
Analyst
PUBLISHED: 02-12-2013
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The first electrochemical immunosensor (EI) for the detection of antibodies against deamidated gliadin peptides (DGP) is described here. A disposable nanohybrid screen-printed carbon electrode modified with DGP was employed as the transducers sensing surface. Real serum samples were successfully assayed and the results were corroborated with an ELISA kit. The presented EI is a promising analytical tool for celiac disease diagnosis.
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Comparison of implant primary stability between maxillary edentulous ridges receiving intramembranous origin block grafts.
Med Oral Patol Oral Cir Bucal
PUBLISHED: 02-05-2013
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The purposes of the present study were: to compare the resonance frequency analysis (RFA) values of implant placed in either ramus or calvaria block grafts; and to determine if implant diameter influences RFA implant stability quotient (ISQ) value.
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Listening in on difficult conversations: an observational, multi-center investigation of real-time conversations in medical oncology.
BMC Cancer
PUBLISHED: 02-04-2013
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The quality of communication in medical care has been shown to influence health outcomes. Cancer patients, a highly diverse population, communicate with their clinical care team in diverse ways over the course of their care trajectory. Whether that communication happens and how effective it is may relate to a variety of factors including the type of cancer and the patients position on the cancer care continuum. Yet, many of the routine needs of cancer patients after initial cancer treatment are often not addressed adequately. Our goal is to identify areas of strength and areas for improvement in cancer communication by investigating real-time cancer consultations in a cross section of patient-clinician interactions at diverse study sites.Methods/designIn this paper we describe the rationale and approach for an ongoing observational study involving three institutions that will utilize quantitative and qualitative methods and employ a short-term longitudinal, prospective follow-up component to investigate decision-making, key topics, and clinician-patient-companion communication dynamics in clinical oncology.
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[Acute coronary syndrome in nonagenarians: Clinical evolution and validation of the main risk scores.]
Rev Esp Geriatr Gerontol
PUBLISHED: 02-03-2013
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Several risk scores regarding the probability of death/complications in the acute setting and during the follow-up of patients admitted with acute coronary syndromes (ACS) have been published, such as the GRACE, TIMI and ZWOLLE risk score. Our objective was to assess the prognosis of nonagenarians admitted to a coronary care unit with an ACS, as well as the usefulness of each of these scores.
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Nafion® modified-screen printed gold electrodes and their carbon nanostructuration for electrochemical sensors applications.
Talanta
PUBLISHED: 01-16-2013
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Screen printed electrodes are frequently used in electroanalytical applications because of their properties such as small size, low detection limit, fast response time, high reproducibility and disposable nature. On the other hand, since the discovery of carbon nanotubes there has been enormous interest in exploring and exploiting their properties, especially for their use in chemical (bio)sensors and nanoscale electronic devices. This paper reports the characterization of gold screen printed electrodes, modified with Nafion(®) and nanostructured with carbon nanotubes and carbon nanofibers dispersed on Nafion(®). The dispersing agent and the nanostructure have a marked effect on the analytical signal that, in turn depends on the intrinsic characteristics of the analyte. Several model analytes have been employed in this study. Anionic, cationic and neutral species such as methylene blue, dopamine, iron (III) sulfate, potassium ferrycianide and urea were considered. The importance for the development of nanostructured sensors relies on the fact that depending on these factors the situation may vary from a notorious enhancement of the signal to a blocking or even decrease.
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Role of 9-lipoxygenase and ?-dioxygenase oxylipin pathways as modulators of local and systemic defense.
Mol Plant
PUBLISHED: 12-22-2011
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Plant 9-lipoxygenases (9-LOX) and ?-dioxygenases (?-DOX) initiate the synthesis of oxylipins after bacterial infection. Here, the role of these enzymes in plants defense was investigated using individual Arabidopsis thaliana lox1 and dox1 mutants and a double lox1 dox1 mutant. Studies with Pseudomonas syringae pv. tomato (Pst) revealed the enhanced susceptibility of lox1 to the virulent strain Pst DC3000 and the partial impairment of lox1 and dox1 mutants to activate systemic acquired resistance. Notably, both defects were enhanced in the lox1 dox1 plants as compared with individual mutants. We found that pre-treatment with 9-LOX- and ?-DOX-generated oxylipins protected plant tissues against bacterial infection. The strongest effect in this respect was exerted by 9-ketooctadecatrienoic acid (9-KOT), which is produced from linolenic acid by 9-LOX. Quantification of 9-KOT revealed its accumulation after bacterial infection. The levels were reduced in lox1 and lox1 dox1 plants but strongly increased in the dox1 mutant due to metabolic interaction of the two pathways. Transcriptional analyses indicated that 9-KOT pre-treatment modifies hormone homeostasis during bacterial infection. The nature of the changes detected suggested that 9-KOT interferes with the hormonal changes caused by bacterial effectors. This notion was substantiated by the finding that 9-KOT failed to reduce the growth of PstDC3000hrpA, a mutant compromised in effector secretion, and of the avirulent strain Pst DC3000 avrRpm1. Further support for the action of the 9-LOX- and ?-DOX-oxylipin pathways as modulators of hormone homeostasis was the observation that lox1 dox1 seedlings are hypersensitive to the growth-inhibitory effect of ABA and showed enhanced activation of ABA-inducible marker genes as compared with wild-type plants.
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A deletion in NRT2.1 attenuates Pseudomonas syringae-induced hormonal perturbation, resulting in primed plant defenses.
Plant Physiol.
PUBLISHED: 12-12-2011
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For an efficient defense response against pathogens, plants must coordinate rapid genetic reprogramming to produce an incompatible interaction. Nitrate Trasnporter2 (NRT2) gene family members are sentinels of nitrate availability. In this study, we present an additional role for NRT2.1 linked to plant resistance against pathogens. This gene antagonizes the priming of plant defenses against the bacterial pathogen Pseudomonas syringae pv tomato DC3000 (Pst). The nrt2 mutant (which is deficient in two genes, NRT2.1 and NRT2.2) displays reduced susceptibility to this bacterium. We demonstrate that modifying environmental conditions that stimulate the derepression of the NRT2.1 gene influences resistance to Pst independently of the total level of endogenous nitrogen. Additionally, hormonal homeostasis seemed to be affected in nrt2, which displays priming of salicylic acid signaling and concomitant irregular functioning of the jasmonic acid and abscisic acid pathways upon infection. Effector-triggered susceptibility and hormonal perturbation by the bacterium seem to be altered in nrt2, probably due to reduced sensitivity to the bacterial phytotoxin coronatine. The main genetic and metabolic targets of coronatine in Arabidopsis (Arabidopsis thaliana) remain largely unstimulated in nrt2 mutants. In addition, a P. syringae strain defective in coronatine synthesis showed the same virulence toward nrt2 as the coronatine-producing strain. Taken together, the reduced susceptibility of nrt2 mutants seems to be a combination of priming of salicylic acid-dependent defenses and reduced sensitivity to the bacterial effector coronatine. These results suggest additional functions for NRT2.1 that may influence plant disease resistance by down-regulating biotic stress defense mechanisms and favoring abiotic stress responses.
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A phase II evaluation of lapatinib in the treatment of persistent or recurrent epithelial ovarian or primary peritoneal carcinoma: a gynecologic oncology group study.
Gynecol. Oncol.
PUBLISHED: 08-12-2011
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Activation and dimerization of the ERBB family play a role in the pathogenesis and progression of ovarian cancer. We conducted a phase II trial to evaluate the activity and tolerability of lapatinib in patients with recurrent or persistent epithelial ovarian cancer (EOC) and to explore the clinical value of expression levels of epidermal growth factor receptors (EGFR), phosphorylated EGFR, HER-2/neu, and Ki-67, and the presence of EGFR mutations.
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Differential clustering of sperm subpopulations in infertile males with clinical varicocele and carriers of rearranged genomes.
J. Androl.
PUBLISHED: 08-11-2011
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Some methods for determining sperm DNA fragmentation, such as the sperm chromatin structure assay (SCSA) and the sperm chromatin dispersion test (SCD), provide additional information about particular subgroups of spermatozoa with specific irregularities. Thus, SCSA recognizes a specific sperm subpopulation, the high-DNA stainability sperm subpopulation (HDS), and SCD recognizes the so-called DNA-degraded sperm (DDS) subpopulation. Although some studies associate the presence of these subpopulations with specific aspects related to infertility, the relationship between both sperm subpopulations and their preponderance in specific clinical groups of infertile males has not been extensively investigated. In this study, HDS and DDS subpopulations were determined in a total of 37 human males: 8 males with proven fertility, 9 infertile males with asthenoteratozoospermia, 10 carriers of chromosomal reorganizations, and 10 infertile males with clinical varicocele. Results showed a significant increase of the DDS subpopulation (P < .001) in both the varicocele patient (16.85 ± 7.24) and carrier of rearranged genome (11.6 ± 5.23) groups, but not in patients with asthenoteratozoospermia (3.88 ± 1.55) or fertile donors (2.62 ± 1.68). No statistical differences were detected for the HDS subpopulation (P = .542), but the highest values were found in the varicocele and rearranged-genome groups. However, no correlation between the HDS and DDS subpopulations were found (r = 0.196; P = .244), suggesting that both represent a different class of sperm subpopulation in the ejaculate. A significant increase in HDS, and especially DDS, can be associated with the presence of varicocele or the rearrangement of chromosomes. Specific diagnostic tests to confirm the diagnosis must be performed in patients with increased DDS and HDS values.
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Celiac disease detection using a transglutaminase electrochemical immunosensor fabricated on nanohybrid screen-printed carbon electrodes.
Biosens Bioelectron
PUBLISHED: 08-03-2011
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Celiac disease is a gluten-induced autoimmune enteropathy characterized by the presence of tissue tranglutaminase (tTG) autoantibodies. A disposable electrochemical immunosensor (EI) for the detection of IgA and IgG type anti-tTG autoantibodies in real patients samples is presented. Screen-printed carbon electrodes (SPCE) nanostructurized with carbon nanotubes and gold nanoparticles were used as the transducer surface. This transducer exhibits the excellent characteristics of carbon-metal nanoparticle hybrid conjugation and led to the amplification of the immunological interaction. The immunosensing strategy consisted of the immobilization of tTG on the nanostructured electrode surface followed by the electrochemical detection of the autoantibodies present in the samples using an alkaline phosphatase (AP) labelled anti-human IgA or IgG antibody. The analytical signal was based on the anodic redissolution of enzymatically generated silver by cyclic voltammetry. The results obtained were corroborated with a commercial ELISA kit indicating that the electrochemical immunosensor is a trustful analytical screening tool.
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Amperometric fructose sensor based on ferrocyanide modified screen-printed carbon electrode.
Talanta
PUBLISHED: 07-20-2011
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The first fructose sensor using a commercial screen-printed ferrocyanide/carbon electrodes (SPFCE) is reported here. The ferrocyanide is included in the carbon ink of the commercial screen-printed carbon electrode. The immobilization of enzyme d-fructose dehydrogenase (FDH) was carried out in an easy way. An aliquot of 10?L FDH was deposited on the electrode surface and left there until dried (approximately 1h) at room temperature. The sensor, so constructed, shows a good sensitivity to fructose (1.25?A/mM) with a slope deviation of ±0.02?A/mM and a linear range comprised between 0.1 and 1mM of fructose, with a limit of detection of 0.05mM. These sensors show good intersensors reproducibility after a previous pretreatment and a high stability. Fructose was determined in real samples as honey, Cola, fruit juices (orange, tomato, apple and pineapple), red wine, red and white grapes, musts and liquor of peach with a good accuracy.
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Discovery of a series of imidazo[4,5-b]pyridines with dual activity at angiotensin II type 1 receptor and peroxisome proliferator-activated receptor-?.
J. Med. Chem.
PUBLISHED: 05-26-2011
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Mining of an in-house collection of angiotensin II type 1 receptor antagonists to identify compounds with activity at the peroxisome proliferator-activated receptor-? (PPAR?) revealed a new series of imidazo[4,5-b]pyridines 2 possessing activity at these two receptors. Early availability of the crystal structure of the lead compound 2a bound to the ligand binding domain of human PPAR? confirmed the mode of interaction of this scaffold to the nuclear receptor and assisted in the optimization of PPAR? activity. Among the new compounds, (S)-3-(5-(2-(1H-tetrazol-5-yl)phenyl)-2,3-dihydro-1H-inden-1-yl)-2-ethyl-5-isobutyl-7-methyl-3H-imidazo[4,5-b]pyridine (2l) was identified as a potent angiotensin II type I receptor blocker (IC(50) = 1.6 nM) with partial PPAR? agonism (EC(50) = 212 nM, 31% max) and oral bioavailability in rat. The dual pharmacology of 2l was demonstrated in animal models of hypertension (SHR) and insulin resistance (ZDF rat). In the SHR, 2l was highly efficacious in lowering blood pressure, while robust lowering of glucose and triglycerides was observed in the male ZDF rat.
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Reciprocal regulation between AtNRT2.1 and AtAMT1.1 expression and the kinetics of NH(4)(+) and NO(3)(-) influxes.
J. Plant Physiol.
PUBLISHED: 05-05-2011
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Our results show that AtNRT2.1 expression has a positive effect on the NH(4)(+) ion influx, mediated by the HATS, as also occurs with AtAMT1.1 expression on the NO(3)(-) ion influx. AtNRT2.1 expression plays a key role in the regulation of AtAMT1.1 expression and in the NH(4)(+) ion influx, differentiating the nitrogen source, and particularly, the lack of it. Nitrogen starvation produces a compensatory effect by AtAMT1.1 when there is an absence of the AtNRT2.1 gene. Our results also show that, in the atnrt2 mutant lacking both AtNRT2.1 and AtNRT2.2, gene functions present different kinetic parameters on the NH(4)(+) ion influx mediated by the HATS, according to the source and availability of nitrogen. Finally, the absence of AMT1.1 also produces changes in the kinetic parameters of the NO(3)(-) influx, showing different V(max) values depending on the source of nitrogen available.
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Transarterial chemoembolization and selective internal radiation for the treatment of patients with metastatic neuroendocrine tumors: a comparison of efficacy and cost.
Oncologist
PUBLISHED: 04-20-2011
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Hepatic arterial therapy (HAT) has been proven to be effective at palliation of hormonal symptoms of metastatic neuroendocrine tumors (NETs), as well as a means of cytoreduction. Recently, the newer modalities of yttrium-90 and drug-eluting beads with doxorubicin (DEBDOX) have been reported to be effective in the treatment of metastatic NETs. The aim of this study was to compare the safety, efficacy, and cost of selective internal radiation with DEB therapy.
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Use of thermogravimetric analysis to monitor the effects of natural laccase mediators on flax pulp.
Bioresour. Technol.
PUBLISHED: 03-18-2011
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The effects of 1-hydroxybenzotriazole (HBT) and the natural laccase mediators gallic acid, caffeic acid and p-hydroxybenzoic acid, on the enzymatic bleaching of flax pulp were compared. The treatment was performed under atmospheric air and oxygen pressure, and, for the first time, monitored by thermogravimetric analysis (TGA) for comparison with chemical analysis, FTIR and X-ray diffraction (XRD) spectroscopies. Thermogravimetric data were simulated by applying a nucleation kinetic equation to a combustion model based on four pseudo-components (hemicellulose, amorphous and crystalline cellulose, and lignin). The results thus obtained show that the effects of the natural mediators are similar to those of HBT and lead to increased order in cellulose microfibril surfaces. An increase in pulp crystallinity was additionally exposed by the XRD and FTIR techniques, and a reduction in microfibril equatorial size by the XRD patterns. Simulated DTG curves were used to determine the kinetic parameters for thermal degradation.
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Activation of the epithelial sodium channel by the metalloprotease meprin ? subunit.
Channels (Austin)
PUBLISHED: 01-01-2011
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The Epithelial Na(+) Channel (ENaC) is an apical heteromeric channel that mediates Na(+) entry into epithelial cells from the luminal cell surface. ENaC is activated by proteases that interact with the channel during biosynthesis or at the extracellular surface. Meprins are cell surface and secreted metalloproteinases of the kidney and intestine. We discovered by affinity chromatography that meprins bind ?-ENaC, a subunit of the ENaC hetero-oligomer. The physical interaction involves NH(2)-terminal cytoplasmic residues 37-54 of ?-ENaC, containing a critical gating domain immediately before the first transmembrane domain, and the cytoplasmic COOH-terminal tail of meprin ? (residues 679-704). This potential association was confirmed by co-expression and co-immunoprecipitation studies. Functional assays revealed that meprins stimulate ENaC expressed exogenously in Xenopus oocytes and endogenously in epithelial cells. Co-expression of ENaC subunits and meprin ? or ?/? in Xenopus oocytes increased amiloride-sensitive Na(+) currents approximately two-fold. This increase was blocked by preincubation with an inhibitor of meprin activity, actinonin. The meprin-mediated increase in ENaC currents in oocytes and epithelial cell monolayers required meprin ?, but not the ? subunit. Meprin ? promoted cleavage of ? and ?-ENaC subunits at sites close to the second transmembrane domain in the extracellular domain of each channel subunit. Thus, meprin ? regulates the activity of ENaC in a metalloprotease-dependent fashion.
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Empirical evaluation of a simple analytical formula for the ultraviolet index.
Photochem. Photobiol.
PUBLISHED: 12-13-2010
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This paper focuses on the estimation of the UV Index (UVI) for all sky conditions using a simple analytical parameterization involving three independent variables: the solar zenith angle, the total ozone column and the clearness index. Measurements of the UVI made at Badajoz and Cáceres (Southwestern Spain) from January 2006 to December 2007 are used to estimate optimal fitting parameters for the model formula, while measurements from January to December 2008 are used to show that the formula-based estimations have mean absolute errors lower than 6% and R(2) ca 0.99.
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The cystic fibrosis transmembrane conductance regulator impedes proteolytic stimulation of the epithelial Na+ channel.
J. Biol. Chem.
PUBLISHED: 08-13-2010
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Cystic fibrosis (CF) is caused by mutations in the CF transmembrane conductance regulator (CFTR) that prevent its proper folding and trafficking to the apical membrane of epithelial cells. Absence of cAMP-mediated Cl(-) secretion in CF airways causes poorly hydrated airway surfaces in CF patients, and this condition is exacerbated by excessive Na(+) absorption. The mechanistic link between missing CFTR and increased Na(+) absorption in airway epithelia has remained elusive, although substantial evidence implicates hyperactivity of the epithelial Na(+) channel (ENaC). ENaC is known to be activated by selective endoproteolysis of the extracellular domains of its ?- and ?-subunits, and it was recently reported that ENaC and CFTR physically associate in mammalian cells. We confirmed this interaction in oocytes by co-immunoprecipitation and found that ENaC associated with wild-type CFTR was protected from proteolytic cleavage and stimulation of open probability. In contrast, ?F508 CFTR, the most common mutant protein in CF patients, failed to protect ENaC from proteolytic cleavage and stimulation. In normal airway epithelial cells, ENaC was contained in the anti-CFTR immunoprecipitate. In CF airway epithelial cultures, the proportion of full-length to total ?-ENaC protein signal was consistently reduced compared with normal cultures. Our results identify limiting proteolytic cleavage of ENaC as a mechanism by which CFTR down-regulates Na(+) absorption.
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The Cav? subunit prevents RFP2-mediated ubiquitination and proteasomal degradation of L-type channels.
Nat. Neurosci.
PUBLISHED: 08-10-2010
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It is well established that the auxiliary Cav? subunit regulates calcium channel density in the plasma membrane, but the cellular mechanism by which this occurs has remained unclear. We found that the Cav? subunit increased membrane expression of Cav1.2 channels by preventing the entry of the channels into the endoplasmic reticulum-associated protein degradation (ERAD) complex. Without Cav?, Cav1.2 channels underwent robust ubiquitination by the RFP2 ubiquitin ligase and interacted with the ERAD complex proteins derlin-1 and p97, culminating in targeting of the channels to the proteasome for degradation. On treatment with the proteasomal inhibitor MG132, Cav?-free channels were rescued from degradation and trafficked to the plasma membrane. The coexpression of Cav? interfered with ubiquitination and targeting of the channel to the ERAD complex, thereby facilitating export from the endoplasmic reticulum and promoting expression on the cell surface. Thus, Cav?? regulates the ubiquitination and stability of the calcium channel complex.
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Phase II trial of bryostatin-1 in combination with cisplatin in patients with recurrent or persistent epithelial ovarian cancer: a California cancer consortium study.
Invest New Drugs
PUBLISHED: 07-21-2010
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The California Cancer Consortium has performed a Phase II trial of infusional bryostatin, a protein kinase C inhibitor isolated from the marine invertebrate bryozoan, Bugula Neritina, a member of the phylum Ectoprocta, in combination with cisplatin, in patients (pts) with recurrent platinum-sensitive or resistant ovarian cancer (OC).
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SPLUNC1 expression reduces surface levels of the epithelial sodium channel (ENaC) in Xenopus laevis oocytes.
Channels (Austin)
PUBLISHED: 07-04-2010
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Throughout the body, the epithelial Na(+) channel (ENaC) plays a critical role in salt and liquid homeostasis. In cystic fibrosis airways, for instance, improper regulation of ENaC results in hyperabsorption of sodium that causes dehydration of airway surface liquid. This dysregulation then contributes to mucus stasis and chronic lung infections. ENaC is known to undergo proteolytic cleavage, which is required for its ability to conduct Na(+) ions. We have previously shown that the short, palate lung and nasal epithelial clone (SPLUNC1) binds to and inhibits ENaC in both airway epithelia and in Xenopus laevis oocytes. In this study, we found that SPLUNC1 was more potent at inhibiting ENaC than either SPLUNC2 or long PLUNC1 (LPLUNC1), two other PLUNC family proteins that are also expressed in airway epithelia. Furthermore, we were able to shed light on the potential mechanism of SPLUNC1s inhibition of ENaC. While SPLUNC1 did not inhibit proteolytic activity of trypsin, it significantly reduced ENaC currents by reducing the number of ENaCs in the plasma membrane. A better understanding of ENaCs regulation by endogenous inhibitors may aid in the development of novel therapies designed to inhibit hyperactive ENaC in cystic fibrosis epithelia.
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Priming for JA-dependent defenses using hexanoic acid is an effective mechanism to protect Arabidopsis against B. cinerea.
J. Plant Physiol.
PUBLISHED: 05-30-2010
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Soil drench treatments with hexanoic acid can effectively protect Arabidopsis plants against Botrytis cinerea through a mechanism based on a stronger and faster accumulation of JA-dependent defenses. Plants impaired in ethylene, salicylic acid, abscisic acid or glutathion pathways showed intact protection by hexanoic acid upon B. cinerea infection. Accordingly, no significant changes in the SA marker gene PR-1 in either the SA or ABA hormone balance were observed in the infected and treated plants. In contrast, the JA signaling pathway showed dramatic changes after hexanoic acid treatment, mainly when the pathogen was present. The impaired JA mutants, jin1-2 and jar1, were unable to display hexanoic acid priming against the necrotroph. In addition, hexanoic acid-treated plants infected with B. cinerea showed priming in the expression of the PDF1.2, PR-4 and VSP1 genes implicated in the JA pathways. Moreover, JA and OPDA levels were primed at early stages by hexanoic acid. Treatments also stimulated increased callose accumulation in response to the pathogen. Although callose accumulation has proved an effective IR mechanism against B. cinerea, it is apparently not essential to express hexanoic acid-induced resistance (HxAc-IR) because the mutant pmr4.1 (callose synthesis defective mutant) is protected by treatment. We recently described how hexanoic acid treatments can protect tomato plants against B. cinerea by stimulating ABA-dependent callose deposition and by priming OPDA and JA-Ile production. We clearly demonstrate here that Hx-IR is a dependent plant species, since this acid protects Arabidopsis plants against the same necrotroph by priming JA-dependent defenses without enhancing callose accumulation.
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Protamine 1 to protamine 2 ratio correlates with dynamic aspects of DNA fragmentation in human sperm.
Fertil. Steril.
PUBLISHED: 04-22-2010
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To investigate the relationship between the protamine 1 to protamine 2 (P1/P2) ratio and the rate of sperm DNA fragmentation in sperm samples from human males with proven fertility and three different cohorts of male patients.
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A phase I trial of dose-dense (biweekly) carboplatin combined with paclitaxel and pegfilgrastim: a feasibility study in patients with untreated Stage III and IV ovarian, tubal or primary peritoneal cancer: a Gynecologic Oncology Group study.
Gynecol. Oncol.
PUBLISHED: 03-19-2010
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Dose-dense regimens have been shown to improve outcome when given as adjuvant therapy to patients with breast cancer compared with their three weekly counterparts. We investigated the feasibility of a dose-dense regimen with carboplatin/paclitaxel followed by pegfilgrastim in patients with advanced ovarian cancer. We also investigated the toxicities including the percentage of patients with grade 2 or greater peripheral neurotoxicity and the clinical response of this regimen.
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Celiac disease diagnosis and gluten-free food analytical control.
Anal Bioanal Chem
PUBLISHED: 02-11-2010
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Celiac disease (CD) is an autoimmune enteropathy, characterized by an inappropriate T-cell-mediated immune response to the ingestion of certain dietary cereal proteins in genetically susceptible individuals. This disorder presents environmental, genetic, and immunological components. CD presents a prevalence of up to 1% in populations of European ancestry, yet a high percentage of cases remain underdiagnosed. The diagnosis and treatment should be made early since untreated disease causes growth retardation and atypical symptoms, like infertility or neurological disorders. The diagnostic criteria for CD, which requires endoscopy with small bowel biopsy, have been changing over the last few decades, especially due to the advent of serological tests with higher sensitivity and specificity. The use of serological markers can be very useful to rule out clinical suspicious cases and also to help monitor the patients, after adherence to a gluten-free diet. Since the current treatment consists of a life-long gluten-free diet, which leads to significant clinical and histological improvement, the standardization of an assay to assess in an unequivocal way gluten in gluten-free foodstuff is of major importance.
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Simulation of the thermogravimetry analysis of three non-wood pulps.
Bioresour. Technol.
PUBLISHED: 01-13-2010
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In a study of three non-wood pulps (rice straw, EFB, and Hesperaloe), the standard pyrolysis model for biomass based on hemicellulose, cellulose, and lignin pseudo-components, was modified to include char volatilization. As a result, abnormally high values for lignin content have been avoided. On the other hand, the consideration of autocatalysis (Prout-Tompkins equation) for TGA data simulation under inert and oxidative atmospheres, allows obtaining a stable set of kinetic parameters that describe volatilization and char oxidation for different heating rates, including char ignition. These simulations allow us to investigate certain effects like oxygen influence on cellulose-char formation (including reduction in activation energy) and to calculate the composition of samples.
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Assessment of the ozone-nitrogen oxide-volatile organic compound sensitivity of Mexico City through an indicator-based approach: measurements and numerical simulations comparison.
J Air Waste Manag Assoc
PUBLISHED: 10-22-2009
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The ozone (O3) sensitivity to nitrogen oxides (NOx, or nitric oxide [NO] + nitrogen dioxide [NO2]) versus volatile organic compounds (VOCs) in the Mexico City metropolitan area (MCMA) is a current issue of scientific controversy. To shed light on this issue, we compared measurements of the indicator species O3/NOy (where NOy represents the sum of NO + NO2 + nitric acid [HNO3] + peroxyacetyl nitrate [PAN] + others), NOy, and the semiempirically derived O3/NOz(surrogate) (where NOz(surrogate) is the derived surrogate NOz, and NOz represents NOx reaction products, or NOy - NOx) with results of numerical predictions reproducing the transition regimes between NOx and VOC sensitivities. Ambient air concentrations of O3, NOx, and NOy were measured from April 14 to 25, 2004 in one downwind receptor site of photochemically aged air masses within Mexico City. MCMA-derived transition values for an episode day occurring during the same monitoring period were obtained through a series of photochemical simulations using the Multiscale Climate and Chemistry Model (MCCM). The comparison between the measured indicator species and the simulated spatial distribution of the indicators O3/ NOy, O3/NOz(surrogate), and NOy in MCMA suggest that O3 in this megacity is likely VOC-sensitive. This is in opposition to past studies that, on the basis of the observed morning VOC/NOx ratios, have concluded that O3 in Mexico City is NOx-sensitive. Simulated MCMA-derived sensitive transition values for O3/NOy, hydrogen peroxide (H2O2)/HNO3, and NOy were found to be in agreement with threshold criteria proposed for other regions in North America and Europe, although the transition crossover for O3/NOz and O3/HNO3 was not consistent with values reported elsewhere. An additional empirical evaluation of weekend/weekday differences in average maximum O3 concentrations and 6:00- to 9:00-a.m. NOx and NO levels registered at the same site in April 2004 indirectly confirmed the above results. A preliminary conclusion is that additional reductions in NOx emissions in MCMA might cause an increase in presently high O3 levels.
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Hexanoic acid-induced resistance against Botrytis cinerea in tomato plants.
Mol. Plant Microbe Interact.
PUBLISHED: 10-09-2009
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We have demonstrated that root treatment with hexanoic acid protects tomato plants against Botrytis cinerea. Hexanoic acid-induced resistance (Hx-IR) was blocked in the jasmonic acid (JA)-insensitive mutant jai1 (a coi1 homolog) and in the abscisic acid (ABA)-deficient mutant flacca (flc). Upon infection, the LoxD gene as well as the oxylipin 12-oxo-phytodienoic acid and the bioactive molecule JA-Ile were clearly induced in treated plants. However, the basal ABA levels were not altered. Hexanoic acid primed callose deposition against B. cinerea in a cultivar-dependent manner. Treated plants from Ailsa Craig, Moneymaker, and Rheinlands Ruhm showed increased callose deposition but not from Castlemart. Hexanoic acid did not prime callose accumulation in flc plants upon B. cinerea infection; therefore, ABA could act as a positive regulator of Hx-IR by enhancing callose deposition. Furthermore, although hexanoic acid protected the JA-deficient mutant defensless1 (def1), the priming for callose was higher than in the wild type. This suggests a link between JA and callose deposition in tomato. Hence, the obtained results support the idea that callose, oxylipins, and the JA-signaling pathway are involved in Hx-IR against B. cinerea. Moreover our data support the relevance of JA-signaling for basal defense against this necrotroph in tomato. Hexanoic acid also protected against Pseudomonas syringae, indicating a broad-spectrum effect for this new inducer.
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Multiple-point electrochemical detection for a dual-channel hybrid PDMS-glass microchip electrophoresis device.
Electrophoresis
PUBLISHED: 10-06-2009
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A new PDMS-based dual-channel MCE with multiple-point amperometric detection has been evaluated. Electrophoresis has been optimised in a single-channel device. Pretreatment with 0.1 M NaOH is very important for increasing and stabilising the EOF. The precision is adequate for a days work in terms of both peak current and migration time. The RSD of the peak current for five successive signals was 1.9, 2.4 and 3.1% for dopamine, p-aminophenol and hydroquinone. RSD for the migration time was always less than 1.3%, which demonstrates the stability of the EOF and the possibility of running multiple experiments in the same microchip. The adequate inter-microchip precision as well as the rapid and simple manufacturing procedure indicates the disposable nature of the PDMS microchips. A dual-channel device with very simple multiple-point amperometric detection is proposed here. Elasticity of the PDMS allows removing the polymer slightly and aligning gold wires working electrodes. Injection can be performed from each of the sample reservoirs or from both simultaneously. The distance between the separation channels is critical for obtaining adequate signals as well as the introduction of a high-voltage electrode in the buffer reservoir. Simultaneous measurement of the same analytes in both channels is possible by applying the same potential. Moreover, since no cross-separation is produced, different analytes or samples can be simultaneously measured.
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[Description of Melolonthidae (Coleoptera) third instar larvae associated to Agave tequilana var. Azul and their population fluctuation in Jalisco, Mexico].
Neotrop. Entomol.
PUBLISHED: 09-02-2009
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Third instars of white grubs of six species associated to agave tequilero in Jalisco, México were described from 1,145 specimens collected from soil samples from September 2006 to August 2007, in the municipalities of Ixtlahuacán del Río, Tepatitlán de Morelos and San Juan de Escobedo, Jalisco, México. Diagnostic characters were illustrated and a key was also included. Cyclocephala comata (Bates) was the most abundant species (63.2%), followed by Phyllophaga ravida (Blanchard) (21.9%), Phyllophaga polyphylla (Bates) (9.4%), Phyllophaga misteca (Bates) (4.1%), Strategus aloeus (L.) (1.1%) and Anomala hoepfneri (Bates) (0.3%). Phyllophaga ravida and A. hoepfneri are reported for the first time on the agave plant and the latter is a new record for the State of Jalisco. All Melolonthidae species showed a marked seasonality with lower number of larvae in June 2007 and high number in August 2007, which is associated with the regions rainy season and the agave plant age, respectively.
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New saponins from Sechium mexicanum.
Magn Reson Chem
PUBLISHED: 07-29-2009
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The chemical study of Sechium mexicanum roots led to the isolation of the two new saponins {3-O-beta-D-glucopyranosyl (1 --> 3)-beta-D-glucopyranosyl-2beta,3beta,16alpha,23-tetrahydroxyolean-12-en-28-oic acid 28-O-alpha-L-rhamnopyranosyl-(1 --> 3)-beta-D-xylopyranosyl-(1 --> 4)-alpha-L-rhamnopyranosyl-(1 --> 2)-alpha-L-arabinopyranoside} (1) and {3-O-beta-D-glucopyranosyl (1 --> 3)-beta-D-glucopyranosyl-2beta,3beta,16alpha,23-tetrahydroxyolean-12-en-28-oic acid 28-O-alpha-L-rhamnopyranosyl-(1 --> 3)-beta-D-xylopyranosyl-(1 --> 4)-[beta-D-apiosyl-(1 --> 3)]-alpha-L-rhamnopyranosyl-(1 --> 2)-alpha-L-arabinopyranoside} (2), together with the known compounds {3-O-beta-D-glucopyranosyl-(1 --> 3)-beta-D-glucopyranosyl-2beta,3beta,6beta,16alpha,23-pentahydroxyolean-12-en-28-oic acid 28-O-alpha-L-rhamnopyranosyl-(1 --> 3)-beta-D-xylopyranosyl-(1 --> 4)-alpha-L-rhamnopyranosyl-(1 --> 2)-alpha-L-arabinopyranoside} (3), tacacosides A(1) (4) and B(3) (5). The structures of saponins 1 and 2 were elucidated using a combination of (1)H and (13)C 1D-NMR, COSY, TOCSY, gHMBC and gHSQC 2D-NMR, and FABMS of the natural compounds and their peracetylated derivates, as well as by chemical degradation. Compounds 1-3 are the first examples of saponins containing polygalacic and 16-hydroxyprotobasic acids found in the genus Sechium, while 4 and 5, which had been characterized partially by NMR, are now characterized in detail.
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Synthesis and evaluation of novel alpha-heteroaryl-phenylpropanoic acid derivatives as PPARalpha/gamma dual agonists.
Bioorg. Med. Chem.
PUBLISHED: 07-16-2009
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The synthesis of a new series of phenylpropanoic acid derivatives incorporating an heteroaryl group at the alpha-position and their evaluation for binding and activation of PPARalpha and PPARgamma are presented in this report. Among the new compounds, (S)-3-{4-[3-(5-methyl-2-phenyl-oxazol-4-yl)-propyl]-phenyl}-2-1,2,3-triazol-2-yl-propionic acid (17j), was identified as a potent human PPARalpha/gamma dual agonist (EC(50)=0.013 and 0.061 microM, respectively) with demonstrated oral bioavailability in rat and dog. 17j was shown to decrease insulin levels, plasma glucose, and triglycerides in the ZDF female rat model. In the human apolipoprotein A-1/CETP transgenic mouse model 17j produced increases in hApoA1 and HDL-C and decreases in plasma triglycerides. The increased potency for binding and activation of both PPAR subtypes observed with 17j when compared to previous analogs in this series was explained based on results derived from crystallographic and modeling studies.
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Fixed-dose rate gemcitabine plus carboplatin in relapsed, platinum-sensitive ovarian cancer patients: results of a three-arm Phase I study.
Gynecol. Oncol.
PUBLISHED: 06-24-2009
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Standard infusion of gemcitabine plus carboplatin showed improved efficacy compared to carboplatin alone in patients with platinum-sensitive (Pt-S) ovarian cancer (OC). Fixed-dose rate (FDR) administration of gemcitabine produces more efficient intracellular phosphorylation of gemcitabine to its active form. This study was designed to identify the maximum tolerated dose (MTD), toxicity profile, and response rate of FDR gemcitabine plus carboplatin in Pt-S OC.
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SPLUNC1 regulates airway surface liquid volume by protecting ENaC from proteolytic cleavage.
Proc. Natl. Acad. Sci. U.S.A.
PUBLISHED: 06-18-2009
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Many epithelia, including the superficial epithelia of the airways, are thought to secrete "volume sensors," which regulate the volume of the mucosal lining fluid. The epithelial Na(+) channel (ENaC) is often the rate limiting factor in fluid absorption, and must be cleaved by extracellular and/or intracellular proteases before it can conduct Na(+) and absorb excess mucosal liquid, a process that can be blocked by proteases inhibitors. In the airways, airway surface liquid dilution or removal activates ENaC. Therefore, we hypothesized that endogenous proteases are membrane-anchored, whereas endogenous proteolysis inhibitors are soluble and can function as airway surface liquid volume sensors to inhibit ENaC activity. Using a proteomic approach, we identified short palate, lung, and nasal epithelial clone (SPLUNC)1 as a candidate volume sensor. Recombinant SPLUNC1 inhibited ENaC activity in both human bronchial epithelial cultures and Xenopus oocytes. Knockdown of SPLUNC1 by shRNA resulted in a failure of bronchial epithelial cultures to regulate ENaC activity and airway surface liquid volume, which was restored by adding recombinant SPLUNC1 to the airway surface liquid. Despite being able to inhibit ENaC, recombinant SPLUNC1 had little effect on extracellular serine protease activity. However, SPLUNC1 specifically bound to ENaC, preventing its cleavage and activation by serine proteases. SPLUNC1 is highly expressed in the airways, as well as in colon and kidney. Thus, we propose that SPLUNC1 is secreted onto mucosal surfaces as a soluble volume sensor whose concentration and dilution can regulate ENaC activity and mucosal volumes, including that of airway surface liquid.
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MCE-electrochemical detection for following interactions of ssDNA and dsDNA with methylene blue.
Electrophoresis
PUBLISHED: 06-12-2009
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The interaction between the organic dye, methylene blue and DNA has been studied by MCE with electrochemical detection. Interaction produces two different signals, one corresponding to free methylene blue and other, for the complex methylene blue-DNA. The hybridization between a ssDNA and a complementary sequence, specific to the severe acute respiratory syndrome virus, has been performed and studied in a thermoplastic olefin polymer of amorphous structure CE-microchip with an end-channel gold wire detector. Moreover, studies with a longer dsDNA, an expression vector involved in the transitory or stable expression in mammals cells, pFLAG-CMV4, has also been performed.
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Ammonium transport and CitAMT1 expression are regulated by N in Citrus plants.
Planta
PUBLISHED: 04-16-2009
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Citrus seedlings (Citrus sinensis L. Osbeck x Poncirus trifoliata Blanco) were used to describe the effects of different N treatments on the NH4+ influx mediated by high- and low-affinity transport systems (HATS and LATS, respectively) and CitAMT1 gene expression. Results show that Citrus plants favor NH4+ over NO3- influx mediated by HATS and LATS when both N sources are present in the nutrient solution and Citrus plants display a much higher capacity to take up NH4+ than NO3-. Furthermore, NH4+ exerts a regulatory effect on NH4+ HATS activity and CitAMT1 expression, both are down-regulated by high N status of the plant, but specifically stimulated by NH4+ and the balance between these two opposite effects depends on the prior nutrition regime of the plant. On the other hand, supply of NO3- inhibits CitAMT1 expression but doesnt affect NH4+ HATS activity on the roots. To explain this discrepancy, it is possible that other CitAMT1 transporters, up-regulated by N limitation, but not repressed by NO3- could be involved in the stimulation of NH4+ HATS activity under pure NO3- nutrition or CitAMT1 transporter could be regulated at the post-transcriptional level.
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Electrochemical study and flow injection analysis of paracetamol in pharmaceutical formulations based on screen-printed electrodes and carbon nanotubes.
Anal. Chim. Acta
PUBLISHED: 02-11-2009
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Acetaminophenol or paracetamol is one of the most commonly used analgesics in pharmaceutical formulations. Acetaminophen is electroactive and voltammetric mechanistic studies for the electrode processes of the acetaminophenol/N-acetyl-p-quinoneimine redox system are presented. Carbon nanotubes modified screen-printed electrodes with enhanced electron transfer properties are used for the study of the electrochemical-chemical oxidation mechanism of paracetamol at pH 2.0. Quantitative analysis of paracetamol by using its oxidation process (in a Britton-Robinson buffer solution pH 10.0) at +0.20 V (vs. an Ag pseudoreference electrode) on an untreated screen-printed carbon electrode (SPCE) was carried out. Thus, a cyclic voltammetric based reproducible determination of acetaminophen (R.S.D., 2.2%) in the range 2.5x10(-6) M to 1x10(-3) M, was obtained. However, when SPCEs are used as amperometric detectors coupled to a flow injection analysis (FIA) system, the detection limit achieved for paracetamol was 1x10(-7) M, one order of magnitude lower than that obtained by voltammetric analysis. The repeatability of the amperometric detection with the same SPCE is 2% for 15 successive injections of 10(-5) M acetaminophen and do not present any memory effect. Finally, the applicability of using screen-printed carbon electrodes for the electrochemical detection of paracetamol (i.e. for quality control analysis) was demonstrated by using two commercial pharmaceutical products.
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Use of autocatalytic kinetics to obtain composition of lignocellulosic materials.
Bioresour. Technol.
PUBLISHED: 01-26-2009
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Non-isothermal thermogravimetric analysis (TGA) data of biomasses and pulps originating from non-wood and alternatives materials (i.e., Tagasaste or rice straw) have been fitted with refined models, which include autocatalytic kinetics. Data sets were obtained for different experimental conditions, such as variations of heating rate and atmosphere, i.e., inert (pyrolysis) versus oxidative atmosphere (combustion). Besides the access to classical kinetic parameters (pre-exponential factor, activation energy, and reaction order), the improved data analysis enabled the determination of the chemical composition of the samples (cellulose, hemicellulose, extractives, lignin). The latter compared very well with those obtained by conventional methods (chemical analysis, HPLC). Given the reduced environmental impact and rapidness of the method, potential applications for research related to new biomasses and industrial processes can be foreseen. The herein implemented method is based on the assumption that samples contain pseudo-components, which independently degrade, and that combustion is the combination of an initial volatilization process (similar to pyrolysis) and a subsequent char oxidation process. Further, it was found that for a reliable modeling of the volatilization stage, extractives should be considered as well, together with the classical pseudo-components: hemicellulose, cellulose and lignin. The char oxidation stage has been simulated as a sum of the oxidation of three char types, one for each main pseudo-component. Importantly, fitting of TGA curves under consideration of autocatalytic kinetics allows the determination of a consistent set of kinetic parameters at different heating rates and leads to significant suppression of the compensation effect. While autocatalysis (characterized by the nucleation order) is not very significant for pyrolysis of biomasses, it can reach high levels for combustion, especially when high heating rates are used. In cellulosic char oxidation a nucleation order larger than one was fitted. The autocatalysis level of the char oxidation can rapidly increase with small modifications of the heating rate (i.e. to pass from 5 to 10 degrees C/min). In this case, the classically applied nth-order kinetic is particularly insufficient to fit experimental data with the same set of the kinetic parameters.
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Simultaneous detection of free and total prostate specific antigen on a screen-printed electrochemical dual sensor.
Biosens Bioelectron
PUBLISHED: 01-23-2009
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Voltammetric enzyme dual sensors for simultaneous determination of free and total prostate specific antigen (fPSA and tPSA) are described. Alkaline Phosphatase (AP) and a mixture solution of 3-indoxyl phosphate and silver ions were used as the enzymatic label and substrate, respectively. 8A6 or 5G6 antibodies specific for free and total PSA, respectively, were immobilized on different screen-printed electrodes (SPEs)--screen-printed carbon electrodes, screen-printed gold electrodes and screen-printed carbon electrodes modified with nanogold--in order to be able to select one of the surfaces as the most adequate one to develop the dual sensor. Screen-printed carbon electrodes modified with nanogold were the SPEs with the best analytical characteristics and lead to the most repeatable bioelectrodes, so they were selected for the development of the dual sensor. On Dualsensor-nAu electrodes, 8A6 antibody was immobilized on one working electrode and 5G6 antibody was immobilized on the other one by deposition of a drop of solution of each antibody and left overnight at 4 degrees C. Biotinylated anti-PSA antibody and streptavidin-AP conjugate were used as detection reagents, giving rise, to our knowledge, to the first simultaneous electrochemical biosensor for free and total PSA. The PSA dual sensor was used to monitor PSA production from three different cultures of human androgen-sensitive prostate tumor cells.
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Exploration of 4,4-disubstituted pyrrolidine-1,2-dicarboxamides as potent, orally active Factor Xa inhibitors with extended duration of action.
Bioorg. Med. Chem.
PUBLISHED: 01-22-2009
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Aiming to improve upon previously disclosed Factor Xa inhibitors, a series of 4,4-disubstituted pyrrolidine-1,2-dicarboxamides were explored with the intent of increasing the projected human half-life versus 5 (projected human t(1/2)=6 h). A stereospecific route to compounds containing a 4-aryl-4-hydroxypyrrolidine scaffold was developed, resulting in several compounds that demonstrated an increase in the half-life as well as an increase in the in vitro potency compared to 5. Reported herein is the discovery of 26, containing a (2R,4S)-4-hydroxy-4-(2,4-difluorophenyl)-pyrrolidine scaffold, which is a selective, orally bioavailable, efficacious Factor Xa inhibitor that appears suitable for a once-daily dosing (projected human t(1/2)=23 h).
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Underivatized polyamine analysis in plant samples by ion pair LC coupled with electrospray tandem mass spectrometry.
Plant Physiol. Biochem.
PUBLISHED: 01-20-2009
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Polyamines are key regulators of cell development and many plant responses to environmental challenges, however, their functions still remain unclear in complex interactions with other hormones and in biotic or abiotic stress. This lack of knowledge derives from the difficulties on measuring natural polyamines in plants. Here, we present a fast multiresidue method for putrescine (Put), 1,3-diaminopropane (DAP), l-ornithine, spermidine (Spd) and spermine (Spn) measurements in plant samples. Polyamine determination is based on a perchloric acid extraction followed by a simple filtration procedure without previous derivatization. Polyamines are resolved by HPLC in a C18 common column and quantified by electrospray ionization tandem mass spectrometry. (13)C(4)-putrescine and 1,7-diaminoheptane standards were added prior to sample extraction to achieve an accurate quantification in a single run. Chromatography of polyamines presents poor retention when reverse phase C18 common columns are used, because they are very polar compounds and contain several positive charges. To circumvent this problem ionic pairing technique has been used successfully with heptafluorobutyric acid (HFBA) at 1mM in the aqueous phase and 25mM in the sample. Improvement of the signal depleted by HFBA has been achieved by adding 1% of propionic acid to the aqueous and organic eluents. All together, gives a method accurate enough to determine polyamines in plants. To demonstrate the usefulness of the method it has been validated in Arabidopsis thaliana samples and polyamines have been determined in several genotypes that over express (35S::ADC2 line 3.6) or are disrupted (adc2) in the Arginine Decarboxylase2 (ADC2) gene.
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Fabrication of SU-8 based microchip electrophoresis with integrated electrochemical detection for neurotransmitters.
Talanta
PUBLISHED: 01-15-2009
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A new SU-8 based microchip capillary electrophoresis (MCE) device has been developed for the first time with integrated electrochemical detection. Embedded electrophoretic microchannels have been fabricated with a multilayer technology based on bonding and releasing steps of stacked SU-8 films. This technology has allowed the monolithic integration in the device of the electrochemical detection system based on platinum electrodes. The fabrication of the chips presented in this work is totally compatible with reel-to-reel techniques, which guarantee a low cost and high reliability production. The influence of relevant experimental variables, such as the separation voltage and detection potential, has been studied on the SU-8 microchip with an attractive analytical performance. Thus, the effective electrical isolation of the end-channel amperometric detector has been also demonstrated. The good performance of the SU-8 device has been proven for separation and detection of the neurotransmitters, dopamine (DA) and epinephrine (EP). High efficiency (30,000-80,000 N/m), excellent precision, good detection limit (450 nM) and resolution (0.90-1.30) has been achieved on the SU-8 microchip. These SU-8 devices have shown a better performance than commercial Topas (thermoplastic olefin polymer of amorphous structure) microchips. The low cost and versatile SU-8 microchip with integrated platinum film electrochemical detector holds great promise for high-volume production of disposable microfluidic analytical devices.
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Hexanoic acid is a resistance inducer that protects tomato plants against Pseudomonas syringae by priming the jasmonic acid and salicylic acid pathways.
Mol. Plant Pathol.
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Hexanoic acid-induced resistance (Hx-IR) is effective against several pathogens in tomato plants. Our study of the mechanisms implicated in Hx-IR against Pseudomonas syringae pv. tomato?DC3000 suggests that hexanoic acid (Hx) treatment counteracts the negative effect of coronatine (COR) and jasmonyl-isoleucine (JA-Ile) on the salicylic acid (SA) pathway. In Hx-treated plants, an increase in the expression of jasmonic acid carboxyl methyltransferase (JMT) and the SA marker genes PR1 and PR5 indicates a boost in this signalling pathway at the expense of a decrease in JA-Ile. Moreover, Hx treatment potentiates 12-oxo-phytodienoic acid accumulation, which suggests that this molecule might play a role per se in Hx-IR. These results support a positive relationship between the SA and JA pathways in Hx-primed plants. Furthermore, one of the mechanisms of virulence mediated by COR is stomatal re-opening on infection with P.?syringae. In this work, we observed that Hx seems to inhibit stomatal opening in?planta in the presence of COR, which suggests that, on infection in tomato, this treatment suppresses effector action to prevent bacterial entry into the mesophyll.
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Design, synthesis, and evaluation of imidazo[4,5-c]pyridin-4-one derivatives with dual activity at angiotensin II type 1 receptor and peroxisome proliferator-activated receptor-?.
Bioorg. Med. Chem. Lett.
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Identification of a series of imidazo[4,5-c]pyridin-4-one derivatives that act as dual angiotensin II type 1 (AT1) receptor antagonists and peroxisome proliferator-activated receptor-? (PPAR?) partial agonists is described. Starting from a known AT1 antagonist template, conformational restriction was introduced by incorporation of an indane ring that when combined with appropriate substitution at the imidazo[4,5-c]pyridin-4-one provided novel series 5 possessing the desired dual activity. The mode of interaction of this series with PPAR? was corroborated through the X-ray crystal structure of 12b bound to the human PPAR? ligand binding domain. Modulation of activity at both receptors through substitution at the pyridone nitrogen led to the identification of potent dual AT1 antagonists/PPAR? partial agonists. Among them, 21b was identified possessing potent dual pharmacology (AT1 IC(50) = 7 nM; PPAR? EC(50) = 295 nM, 27% max) and good ADME properties.
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