Home About us Contact | |||
Time-course Analysis (time-course + analysis)
Selected AbstractsAcute Blood Pressure Changes After the Onset of Atrioventricular Nodal Reentrant Tachycardia: A Time-Course AnalysisJOURNAL OF CARDIOVASCULAR ELECTROPHYSIOLOGY, Issue 10 2005MEHDI RAZAVI M.D. Introduction: We aimed to characterize blood pressure (BP) response at the beginning of atrioventricular nodal reentrant tachycardia (AVNRT) and its relationship to orthostatic challenge and variable atrioventricular interval. Methods and Results: In this prospective study of 17 consecutive patients with documented AVNRT, mean BP was analyzed in the supine and upright positions during sinus rhythm, AVNRT, and pacing with atrioventricular delay of 150 msec (AV150) and 0 msec (AV0). Mean BPs were compared at 3,5 seconds, 8,10 seconds, and 28,30 seconds after the onset of AVNRT or pacing. BP decreased immediately after AVNRT initiation, with gradual recovery during the first 30 seconds from 71.9 ± 16.5 mmHg to 86 ± 13.8 mmHg, P < 0.01. A similar pattern was observed during AV0, but not during AV150, pacing. While supine, mean BP decrease was more pronounced during AVNRT and AV0 pacing (,26.1% and ,32.1%, respectively) than during AV150 pacing (,8%, P = 0.02 and P = 0.07, respectively). This difference subsided 30 seconds after the onset of AVNRT or pacing. When upright, the mean BP time course was similar, but mean BP recovery during AVNRT was slower, and the difference between mean BP during AVNRT and AV150 persisted at 30 seconds. Conclusions: The initial mean BP decrease during AVNRT recovered gradually within 30 seconds. A short atrioventricular interval is associated with a greater mean BP decrease at the onset of tachycardia. These observations may explain clinical symptoms immediately after the onset of AVNRT. [source] S25.4: Time-course Analysis in Crossover TrialsBIOMETRICAL JOURNAL, Issue S1 2004Berthold Schneider No abstract is available for this article. [source] Carbon Monoxide Alleviates Salt-Induced Oxidative Damage in Wheat Seedling LeavesJOURNAL OF INTEGRATIVE PLANT BIOLOGY, Issue 3 2006Ben-Kai Huang Abstract Carbon monoxide (CO), a by-product released during the degradation of heme by heme oxygenases (EC 1.14.99.3) in animals, is regarded as an important physiological messenger or bioactive molecule involved in many biological events that has been recently reported as playing a major role in mediating the cytoprotection against oxidant-induced lung injury. In the present study, we first determined the protective effect of exogenous CO against salt-induced oxidative damage in wheat seedling leaves. Wheat seedlings treated with 0.01 ,mol/L hematin as the CO donor demonstrated significant reversal of chlorophyll decay, dry weight, and water loss induced by 300 mmol/L NaCl stress. Interestingly, the increase in lipid peroxidation observed in salt-treated leaves was reversed by 0.01 nmol/L hematin treatment. Time-course analyses showed that application of 0.01 ,mol/L hematin enhanced guaiacol peroxidase, superoxide dismutase, ascorbate peroxidase and catalase activities in wheat seedling leaves subjected to salt stress. These effects are specific for CO because the CO scavenger hemoglobin (1.2 mg/L) blocked the actions of the CO donor hematin. However, higher concentration of the CO donor (1.0 ,mol/L) did not alleviate dry weight and water loss of salt-stressed wheat seedlings. These results suggest that exogenous application of low levels of a CO donor may be advantageous against salinity toxicity. (Managing editor: Ping He) [source] Long-lasting coexpression of nestin and glial fibrillary acidic protein in primary cultures of astroglial cells with a major participation of nestin+/GFAP, cells in cell proliferationJOURNAL OF NEUROSCIENCE RESEARCH, Issue 8 2006Solène Sergent-Tanguy Abstract Nestin, a currently used marker of neural stem cells, is transiently coexpressed with glial fibrillary acidic protein (GFAP) during development and is induced in reactive astrocytes following brain injury. Nestin expression has also been found in cultures of astroglial cells, but little is known about the fate and the mitotic activity of nestin-expressing cells in this in vitro model. The present study reveals a long-lasting expression of nestin in primary cultures of astroglial cells derived from the rat brain. Over 70% of the cells were nestin+ at 12 weeks, with a large majority coexpressing the GFAP astrocytic marker. Time-course analyses supported a transition from a nestin+/GFAP, to a nestin+/GFAP+ phenotype over time, which was further increased by cell cycle arrest. Interestingly, double staining with Ki67 revealed that over 90% of cycling cells were nestin+ whereas only 28% were GFAP+ in a population consisting of almost equivalent numbers of nestin+ and GFAP+ cells. These observations indicated that nestin+/GFAP, cells are actively engaged in mitotic activity, even after 2 weeks in vitro. Part of these cells might have retained properties of neural stem cells, insofar as 10% of cells in a primary culture of glial cells were able to generate neurospheres that gave rise to both neurons and astrocytes. Further studies will be necessary to characterize fully the proliferating cells in primary cultures of glial cells, but our present results reveal a major contribution of the nestin+/GFAP, cells to the increase in the number of astrocytes, even though nestin+/GFAP+ cells proliferate also. © 2006 Wiley-Liss, Inc. [source] Cancer biomarker discovery via low molecular weight serum proteome profiling , Where is the tumor?PROTEOMICS - CLINICAL APPLICATIONS, Issue 12 2007Michael T. Davis Abstract Time-course analyses of rapidly processed serum performed in parallel by SELDI and nanoscale LC-MS/MS have revealed the temporal correlation of several literature-based disease markers with ex vivo driven events such that their in vivo existence in healthy subjects is questionable. Identification by MS/MS reveals these putative biomarkers to be byproducts of the coagulation cascade and platelet activation and suggests plasmatic analysis may be preferred. In a pilot plasmatic study, a cohort of naïve prostate cancer (PCa) samples were uniformly distinguished from their age-matched controls (n,=,20) on the basis of multiple peptidic components; most notably by a derivative of complement C4 at 1863,m/z (GLEEELQFSLGSKINVK, C41353,1369). The fully tryptic nature of this and other putative PCa discriminants is consistent with the cleavage specificity of common blood proteases and questions the need for tumor-derived proteolytic activities as has been proposed. In light of the known correlation of disregulated hemostasis with malignant disease, we suggest the underlying differentiating phenomena in these types of analyses may lie in the temporal disparity of sample activation such that the case (patient) samples are preactivated while the control samples are not. [source] Osteoclastogenesis-Related Antigen, a Novel Molecule on Mouse Stromal Cells, Regulates Osteoclastogenesis,JOURNAL OF BONE AND MINERAL RESEARCH, Issue 4 2003Satoshi Arai Abstract Osteoclastogenesis is regulated by RANKL expressed on stromal cells. In this study, we sought to isolate a new surface molecule regulating osteoclastogenesis on stromal cells by generating monoclonal antibodies. A rat was immunized with the mouse stromal cell line, TSB13, which can support osteoclastogenesis, and a monoclonal antibody, A15-1, was obtained. A15-1 bound to a surface antigen on TSB13 cells, termed osteoclastogenesis-related antigen (OCRA), and immunoprecipitation with this antibody revealed that OCRA was a 220-kDa molecule. By means of flow cytometry, the A15-1 antigen (OCRA) was found to be expressed on various mesenchymal cell lines but not on hematopoietic cell lines, and the expression level of OCRA on the TSB13 cells was slightly increased by treatment with 1,,25(OH)2D3. When osteoclast progenitors and TSB13 cells were co-cultured in the presence of 1,,25(OH)2D3, the addition of A15-1 inhibited osteoclast differentiation in a dose-dependent manner; however, no significant inhibition of soluble RANKL-induced osteoclastogenesis was observed, suggesting that A15-1 inhibited only stromal cell-dependent osteoclastogenesis. The same inhibitory effect of A15-1 was also observed when primary bone marrow-derived stromal cells were used. The osteoclastogenesis-promoting effects of other osteotropic factors, such as parathyroid hormone (PTH) and interleukin (IL)-1,, were also inhibited by A15-1. Time-course analysis of osteoclast differentiation in vitro indicated that the initial 2 days of treatment with A15-1 was sufficient for inhibition, suggesting that A15-1 inhibits the early stages of osteoclast differentiation. Finally, we investigated the in vivo effects of A15-1 on PTH-induced hypercalcemia in mice. Treatment with A15-1 significantly decreased the osteoclast surface in the PTH-administered mice. Taken together, our data indicate that OCRA, a novel A15-1-detected antigen, regulates stromal cell-dependent osteoclastogenesis. [source] Differential control of apoptosis by DJ-1 in prostate benign and cancer cellsJOURNAL OF CELLULAR BIOCHEMISTRY, Issue 6 2004Yaacov Hod Abstract DJ-1 is a conserved protein reported to be involved in diverse cellular processes ranging from cellular transformation, control of protein,RNA interaction, oxidative stress response to control of male infertility, among several others. Mutations in the human gene have been shown to be associated with an autosomal recessive, early onset Parkinson's disease (PARK7). The present study examines the control of DJ-1 expression in prostatic benign hyperplasia (BPH-1) and cancer (PC-3) cell lines in which DJ-1 abundance differs significantly. We show that while BPH-1 cells exhibit low basal level of DJ-1 expression, stress-inducing agents such as H2O2 and mitomycin C markedly increase the intracellular level of the polypeptide. In contrast, DJ-1 expression is relatively high in PC-3 cells, and incubation with the same cytotoxic drugs does not modulate further the level of the polypeptide. In correlation with the expression of DJ-1, both cytotoxic agents activate the apoptotic pathway in the prostatic benign cells but not in PC-3 cells, which are resistant to their action. We further demonstrate that incubation of BPH-1 cells with TNF-related-apoptosis-inducing-ligand/Apo-2L (TRAIL) also enhances DJ-1 expression and that TRAIL and H2O2 act additively to stimulate DJ-1 accumulation but synergistically in the activation of the apoptotic pathway. Time-course analysis of DJ-1 stimulation shows that while DJ-1 level increases without significant lag in TRAIL-treated cells, there is a delay in H2O2 -treated cells, and that the increase in DJ-1 abundance precedes the activation of apoptosis. Unexpectedly, over-expression of DJ-1 de-sensitizes BPH-1 cells to the action of apoptotic-inducing agents. However, RNA-interference-mediated silencing of DJ-1 expression results in sensitization of PC-3 cells to TRAIL action. These results are consistent with a model in which DJ-1 is involved in the control of cell death in prostate cell lines. DJ-1 appears to play a differential role between cells expressing a low but inducible level of DJ-1 (e.g., BPH-1 cells) and those expressing a high but constitutive level of the polypeptide (e.g., PC-3 cells). © 2004 Wiley-Liss, Inc. [source] Amplification of low quantity bacterial RNA for microarray studies: time-course analysis of Leptospirillum ferrooxidans under nitrogen-fixing conditions,ENVIRONMENTAL MICROBIOLOGY, Issue 6 2006Mercedes Moreno-Paz Summary We have developed a method for the amplification of low quantity total bacterial RNA for DNA microarrays analysis. Current methods are based on the linear amplification by the in vitro transcription from the T7 promoter, similar to that used for eukaryotic mRNA amplification. For the incorporation of T7 promoter, the prokaryotic RNA must be enzymatically modified for the incorporation of a polyA tail at the 3, end to emulate the eukaryotic mRNA. The method we describe and validate herein avoids this step by the direct and random incorporation of the T7 promoter. From 500 ng of total bacterial RNA, we obtained 130,150 µg of antisense RNA, such products being good substrate for fluorescent labelling and DNA microarray analysis. The method was validated with bacterial samples from which it is very difficult to obtain sufficient amounts and quality of total RNA for global gene expression analysis. This is critical for low cell density growing microorganisms, environmental samples, or many extremophiles where the composition of the cultural media severely affects the RNA yield, like in the case of the acidophile and iron oxidizer Gram-negative bacterium Leptospirillum ferrooxidans. We further validated our amplification method in parallel experiments with non-amplified RNA by following the expression of the L. ferrooxidans nif regulon along the time-course of growth. [source] Benzidine transformation processes in natural sedimentsENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 8 2006Joel Harden Abstract Aromatic amines, such as benzidine and 3,3,-dichlorobenzidine, are chemicals used in the pigment and dye processes. Release of these compounds into the environment is important because of their carcinogenic and toxic nature. In the present study, the sediment and water samples were collected from Lake Macatawa (Holland, MI, USA) and subsequently spiked with benzidine. The grain size distribution of the sediment samples investigated here ranged in composition from sandy to silty-clay sediment types. The sediment,water systems spiked with benzidine were incubated under anaerobic conditions at 4, 15, and 23°C for 211 d. Degradation of benzidine was observed over the time-course analysis of the sediment,water mixtures. Three possible metabolites (aniline, 2-ethyl-1-hexanol, and 1-amino-2-hexene) were observed during this investigation as a result of gas chromatography/mass spectrometry and liquid chromatography/mass spectrometry. No metabolites were observed in autoclaved bottles, suggesting that the transformation of benzidine in the sediment,water mixtures was the result of microbial activity. From sediment,water distribution experiments, benzidine demonstrated higher sorption affinity for the different sediment phases than its degradation product, aniline. Therefore, microbially mediated transformation of benzidine to aniline is expected to yield a greater total concentration of the more mobile compound, aniline, in the water phase and a greater possibility for transport of aniline in the water phase. [source] Catalase inhibition alters suberization and wound healing in potato (Solanum tuberosum) tubersPHYSIOLOGIA PLANTARUM, Issue 3 2007Mohammed Bajji In response to wounding, potato (Solanum tuberosum L.) tubers generate hydrogen peroxide (H2O2) in association with suberization, a critical phase of the wound-healing process. In the present study, the effect of aminotriazole (AT), a catalase (CAT, EC 1.11.1.6) inhibitor, on cut tubers was investigated using fresh weight (FW) loss and pathogen attack symptoms as indicators of wound-healing efficiency. Seven days after treatment, AT-treated tuber halves lost more FW and developed infection signs compared with the controls. Thiourea, another CAT inhibitor, as well as exogenous H2O2 treatments induced the same effects as AT suggesting that the alteration of the wound healing may be caused by CAT inhibition and the resulting accumulation of H2O2. Using transgenic tubers, FW losses 1 week after wounding were either higher (CAT repression) or lower (CAT overexpression) than those of the wild-type. When tuber halves were allowed to wound heal for different periods before treatment, AT had no effect on the progress of their wound healing if wound-healed for at least 3 days. This implies that AT may affect early wound-healing-related events, especially those occurring before or during suberization. A time-course analysis of the effects of AT treatment on wounded tuber tissues revealed that AT prevented the deposition of the polyphenolic domain of suberin in association with CAT inhibition and H2O2 accumulation. These data are important in identifying factors that may be required to regulate suberization and contribute to a better understanding of this critical process to hasten its rate and limit wound-related losses in stored potato tubers. [source] Control of misincorporation of serine for asparagine during antibody production using CHO cellsBIOTECHNOLOGY & BIOENGINEERING, Issue 1 2010Anurag Khetan Abstract A recombinant monoclonal antibody produced by Chinese hamster ovary (CHO) cell fed-batch culture was found to have amino acid sequence misincorporation upon analysis by intact mass and peptide mapping mass spectrometry. A detailed analysis revealed multiple sites for asparagine were being randomly substituted by serine, pointing to mistranslation as the likely source. Results from time-course analysis of cell culture suggest that misincorporation was occurring midway through the fed-batch process and was correlated to asparagine reduction to below detectable levels in the culture. Separate shake flask experiments were carried out that confirmed starvation of asparagine and not excess of serine in the medium as the root cause of the phenomenon. Reduction in serine concentration under asparagine starvation conditions helped reduce extent of misincorporation. Supplementation with glutamine also helped reduce extent of misincorporation. Maintenance of asparagine at low levels in 2,L bench-scale culture via controlled supplementation of asparagine-containing feed eliminated the occurrence of misincorporation. This strategy was implemented in a clinical manufacturing process and scaled up successfully to the 200 and 2,000,L bioreactor scales. Biotechnol. Bioeng. 2010;107: 116,123. © 2010 Wiley Periodicals, Inc. [source] |