248 research outputs found

    Technical Note: Field experiences using UV/VIS sensors for high-resolution monitoring of nitrate in groundwater

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    peer-reviewedTwo different in situ spectrophotometers are compared that were used in the field to determine nitrate-nitrogen (NO3-N) concentrations at two distinct spring discharge sites. One sensor was a double wavelength spectrophotometer (DWS) and the other a multiple wavelength spectrophotometer (MWS). The objective of the study was to review the hardware options, determine ease of calibration, accuracy, influence of additional substances and to assess positive and negative aspects of the two sensors as well as troubleshooting and trade-offs. Both sensors are sufficient to monitor highly time-resolved NO3-N concentrations in emergent groundwater. However, the chosen path length of the sensors had a significant influence on the sensitivity and the range of detectable NO3-N. The accuracy of the calculated NO3-N concentrations of the sensors can be affected if the content of additional substances such as turbidity, organic matter, nitrite or hydrogen carbonate significantly varies after the sensors have been calibrated to a particular water matrix. The MWS offers more possibilities for calibration and error detection but requires more expertise compared with the DWS.The authors would like to acknowledge the Teagasc Walsh Fellowship scheme for funding the study in Ireland, and the German federal Ministry of Education and Research (BMBF) for sponsoring the SMART-project (grant no. 02WM1079-1086, 02WM1211-1212) for the study in Jordan.Teagasc Walsh Fellowship Programm

    Assorted effects of TGFβ and chondroitinsulfate on p38 and ERK1/2 activation levels in human articular chondrocytes stimulated with LPS

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    SummaryObjectivesInadequate cellular response of chondrocytes to stress frequently terminates in osteoarthritis (OA). Adequate response is fundamentally modulated by concerted cytokine signaling events, directing degradation and synthesis of cartilage on articular surfaces where and whenever necessary. Transforming growth factor (TGF)β is a prominent mediator in cartilage anabolism, although particular catabolic activities are occasionally reported. Clearly, before the TGFβ signal gets through to the gene regulatory machinery, cross talk with modulators occurs.MethodWe tested the hypothesis whether chondroitinsulfate (CS) modulates cell signaling. TGFβ and/or soluble CS was added to human articular chondrocytes (HACs) and activation of p38 and extracellular signal related kinase (ERK)1/2 was determined by immunoblot analysis. Expression levels of mRNA of matrix metalloproteinase (MMP)-2, -3 and -13 were determined by real-time polymerase chain reaction (PCR).ResultsNo significant effects were observed unless cells were stimulated with lipopolysaccharide (LPS), invigorating catabolic metabolism in chondrocytes. LPS effects, however, were profoundly modulated by TGFβ, CS and both applied in combination. Most prominent, the silencing of p38 stress signal by CS was superimposable to that of TGFβ. Phospho-ERK1/2 levels were raised by TGFβ three-fold over LPS induced levels. In contrast, CS treatment, alone or combined with TGFβ, reduced phosphorylation significantly below LPS induced levels. Finally, suppression of LPS induced MMP-13 mRNA levels resulted with CS.ConclusionSoluble CS modulates signaling events in chondrocytes concurrent with MMP-13 down regulation. The effects observed suggest a feedback signaling mechanism cross talking with TGFβ-signal pathways and may serve an explanation, on the cellular level, for the beneficial effects found in clinical studies with pharmacologic application of CS

    WormBase 2007

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    WormBase (www.wormbase.org) is the major publicly available database of information about Caenorhabditis elegans, an important system for basic biological and biomedical research. Derived from the initial ACeDB database of C. elegans genetic and sequence information, WormBase now includes the genomic, anatomical and functional information about C. elegans, other Caenorhabditis species and other nematodes. As such, it is a crucial resource not only for C. elegans biologists but the larger biomedical and bioinformatics communities. Coverage of core areas of C. elegans biology will allow the biomedical community to make full use of the results of intensive molecular genetic analysis and functional genomic studies of this organism. Improved search and display tools, wider cross-species comparisons and extended ontologies are some of the features that will help scientists extend their research and take advantage of other nematode species genome sequences

    On-Chip Contactless Four-Electrode Conductivity Detection for Capillary Electrophoresis Devices

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    In this contribution, a capillary electrophoresis microdevice with an integrated on-chip contactless fourelectrode conductivity detector is presented. A 6-cm-long, 70-µm-wide, and 20-µm-deep channel was etched in a glass substrate that was bonded to a second glass substrate in order to form a sealed channel. Four contactless electrodes (metal electrodes covered by 30-nm silicon carbide) were deposited and patterned on the second glass substrate for on-chip conductivity detection. Contactless conductivity detection was performed in either a two-or a four-electrode configuration. Experimental results confirmed the improved characteristics of the fourelectrode configuration over the classical two-electrode detection setup. The four-electrode configuration allows for sensitive detection for varying carrier-electrolyte background conductivity without the need for adjustment of the measurement frequency. Reproducible electrophoretic separations of three inorganic cations (K + , Na + , Li + ) and six organic acids are presented. Detection as low as 5 µM for potassium was demonstrated. In the development and optimization of miniaturized analytical systems, a delicate combination of science and technology originating from microelectronic device fabrication, electrical engineering, and analytical chemistry is essential. In this multidisciplinary field, microtechnology experts combine the demands from analytical chemistry and electronic instrumentation in the design and fabrication of novel analytical devices. 1,2 Chemical analysis systems, such as high-performance liquid chromatography (HPLC) or capillary electrophoresis (CE), always consist of the combination of a separation and a detection system. For separation, CE or CE-based separation techniques are highly suitable for implementation on the microchip format. Electrokinetic control of fluid transport eliminates the need for external components such as pumps and valves. The separation efficiency is relatively independent of the separation path length and is, therefore, more compatible with miniaturization than, for instance, chromatographic techniques. As far as detection is concerned, laser-induced fluorescence (LIF) is, at present, the most widely used detection technique in miniaturized analysis systems because of its high sensitivity. The drawbacks of LIF are its limited compatibility with miniaturization and on-chip integration and the requirement for labeling of most (bio) chemically relevant compounds. External devices such as the relatively large laser and the photodetector system strongly prohibit further miniaturization. The development of alternative detection methods compatible with miniaturization and full onchip integration is highly desirable. Since electrode deposition is a well-established process in microfabrication, the implementation of detection techniques utilizing integrated electrodes has become an attractive approach. Successful coupling of conventional CE with potentiometry, 3 amperometry, 4,5 and conductometry 6-10 has been reported in the literature. In addition, both amperometric and potentiometric detection were also implemented in chip-based CE systems. [11][12][13] The primary advantage of amperometric and potentiometric detection over conductivity detection is the high selectivity induced by the electrochemical reactions that take place at the electrode surface. Only electrochemically active compounds * Corresponding author: (tel) +31 (0) 15 278 6518; (fax) +31 (0) 15 278 5755

    Cell-surface residence of sphingosine 1-phosphate receptor 1 on lymphocytes determines lymphocyte egress kinetics

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    The sphingosine 1-phosphate receptor 1 (S1P1) promotes lymphocyte egress from lymphoid organs. Previous work showed that agonist-induced internalization of this G protein–coupled receptor correlates with inhibition of lymphocyte egress and results in lymphopenia. However, it is unclear if S1P1 internalization is necessary for this effect. We characterize a knockin mouse (S1p1rS5A/S5A) in which the C-terminal serine-rich S1P1 motif, which is important for S1P1 internalization but dispensable for S1P1 signaling, is mutated. T cells expressing the mutant S1P1 showed delayed S1P1 internalization and defective desensitization after agonist stimulation. Mutant mice exhibited significantly delayed lymphopenia after S1P1 agonist administration or disruption of the vascular S1P gradient. Adoptive transfer experiments demonstrated that mutant S1P1 expression in lymphocytes, rather than endothelial cells, facilitated this delay in lymphopenia. Thus, cell-surface residency of S1P1 on T cells is a primary determinant of lymphocyte egress kinetics in vivo

    Experimental identification and characterization of 97 novel npcRNA candidates in Salmonella enterica serovar Typhi

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    We experimentally identified and characterized 97 novel, non-protein-coding RNA candidates (npcRNAs) from the human pathogen Salmonella enterica serovar Typhi (hereafter referred to as S. typhi). Three were specific to S. typhi, 22 were restricted to Salmonella species and 33 were differentially expressed during S. typhi growth. We also identified Salmonella Pathogenicity Island-derived npcRNAs that might be involved in regulatory mechanisms of virulence, antibiotic resistance and pathogenic specificity of S. typhi. An in-depth characterization of S. typhi StyR-3 npcRNA showed that it specifically interacts with RamR, the transcriptional repressor of the ramA gene, which is involved in the multidrug resistance (MDR) of Salmonella. StyR-3 interfered with RamR–DNA binding activity and thus potentially plays a role in regulating ramA gene expression, resulting in the MDR phenotype. Our study also revealed a large number of cis-encoded antisense npcRNA candidates, supporting previous observations of global sense–antisense regulatory networks in bacteria. Finally, at least six of the npcRNA candidates interacted with the S. typhi Hfq protein, supporting an important role of Hfq in npcRNA networks. This study points to novel functional npcRNA candidates potentially involved in various regulatory roles including the pathogenicity of S. typhi
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