TY - JOUR A1 - Dongmo, Saustin A1 - Leyk, Janina A1 - Dosche, Carsten A1 - Richter-Landsberg, Christiane A1 - Wollenberger, Ursula A1 - Wittstock, Gunther T1 - Electrogeneration of O-2(center dot-) and H2O2 Using Polymer-modified Microelectrodes in the Environment of Living Cells JF - Electroanalysis : an international journal devoted to fundamental and practical aspects of electroanalysis N2 - Microelectrodes modified with electropolymerized plumbagin (PLG) were used for the generation of superoxide radical (O-2(center dot-)) and hydrogen peroxide (H2O2) during oxygen reduction reaction (ORR) in an aqueous medium, specifically in serum-free cell culture media. This is enabled by the specific design of a polymer film on the microelectrode. The generation and diffusion of O-2(center dot-) during electrocatalytic ORR at a positionable PLG polymer-modified microelectrode was followed by fluorescence microscopy with the selective dye 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl) and by amperometric detection using a cytochrome c-modified electrode at + 0.13 V. H2O2 production, either by direct oxygen reduction or as product of O-2(center dot-) disproportionation, was monitored by the reaction with Amplex UltraRed. The PLG polymer-modified microelectrodes were used to expose mammalian B6-RPE07 retinal cells to defined local fluxes of reactive oxygen species (ROS), and cellular responses and morphological alterations were observed. The use of a controllable source of ROS opens many possibilities to study how living cells respond to the presence of a certain flux of specific ROS. KW - reactive oxygen species KW - microelectrode KW - scanning electrochemical microscopy KW - biosensor KW - polymer-modified electrode KW - oxygen reduction reaction Y1 - 2016 U6 - https://doi.org/10.1002/elan.201600267 SN - 1040-0397 SN - 1521-4109 VL - 28 SP - 2400 EP - 2407 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Riebe, Daniel A1 - Beitz, Toralf A1 - Dosche, Carsten A1 - Löhmannsröben, Hans-Gerd A1 - Raab, Volker A1 - Raab, Corinna A1 - Unverzagt, Matthias T1 - High-resolution spectrometer using combined dispersive and interferometric wavelength separation for raman and laser-induced Breakdown Spectroscopy (LIBS) JF - Applied spectroscopy : an international journal of spectroscopy ; official publication of the Society for Applied Spectroscopy N2 - In this paper the concept of a compact high-resolution spectrometer based on the combination of dispersive and interferometric elements is presented. Dispersive elements are used to spectrally resolve the light in one direction with coarse resolution (Delta lambda < 0.5 nm), while perpendicular to that direction an etalon provides high spectral resolution (Delta lambda < 50 pm). This concept for two-dimensional spectroscopy has been implemented for the wavelength range lambda = 350-650 nm. Appropriate algorithms for reconstructing spectra from the two-dimensional raw data and for wavelength calibration were established in an analysis software. Potential applications for this new spectrometer are Raman and laser-induced breakdown spectroscopy (LIBS). Resolutions down to 28 pm (routinely 54 pm) could be realized for these applications. KW - Raman spectroscopy KW - Laser-induced breakdown spectroscopy KW - LIBS KW - Fabry-Perot etalon KW - High-resolution spectrometer Y1 - 2014 U6 - https://doi.org/10.1366/13-07426 SN - 0003-7028 SN - 1943-3530 VL - 68 IS - 9 SP - 1030 EP - 1038 PB - Society for Applied Spectroscopy CY - Frederick ER - TY - JOUR A1 - Zude, Manuela A1 - Pflanz, Michael A1 - Spinelli, Lorenzo A1 - Dosche, Carsten A1 - Torricelli, Alessandro T1 - Non-destructive analysis of anthocyanins in cherries by means of Lambert-Beer and multivariate regression based on spectroscopy and scatter correction using time-resolved analysis JF - Journal of food engineering N2 - In high-value sweet cherry (Prunus avium), the red coloration - determined by the anthocyanins content - is correlated with the fruit ripeness stage and market value. Non-destructive spectroscopy has been introduced in practice and may be utilized as a tool to assess the fruit pigments in the supply chain processes. From the fruit spectrum in the visible (Vis) wavelength range, the pigment contents are analyzed separately at their specific absorbance wavelengths. A drawback of the method is the need for re-calibration due to varying optical properties of the fruit tissue. In order to correct for the scattering differences, most often the spectral intensity in the visible spectrum is normalized by wavelengths in the near infrared (NIR) range, or pre-processing methods are applied in multivariate calibrations. In the present study, the influence of the fruit scattering properties on the Vis/NIR fruit spectrum were corrected by the effective pathlength in the fruit tissue obtained from time-resolved readings of the distribution of time-of-flight (DTOF). Pigment analysis was carried out according to Lambert-Beer law, considering fruit spectral intensities, effective pathlength, and refractive index. Results were compared to commonly applied linear color and multivariate partial least squares (PLS) regression analysis. The approaches were validated on fruits at different ripeness stages, providing variation in the scattering coefficient and refractive index exceeding the calibration sample set. In the validation, the measuring uncertainty of non-destructively analyzing fruits with Vis/NIR spectra by means of PLS or Lambert-Beer in comparison with combined application of Vis/NIR spectroscopy and DTOF measurements showed a dramatic bias reduction as well as enhanced coefficients of determination when using both, the spectral intensities and apparent information on the scattering influence by means of DTOF readings. Corrections for the refractive index did not render improved results. KW - Cherry KW - DTOF KW - Effective pathlength KW - Fruit maturity KW - Lambert-Beer KW - NIR KW - Non-invasive KW - Pigments KW - PLS KW - Ripeness KW - Sensor fusion KW - Spectroscopy KW - Time-resolved spectroscopy KW - TIRF KW - Vis KW - Scattering Y1 - 2011 U6 - https://doi.org/10.1016/j.jfoodeng.2010.09.021 SN - 0260-8774 VL - 103 IS - 1 SP - 68 EP - 75 PB - Elsevier CY - Oxford ER - TY - JOUR A1 - Borwankar, Tejas A1 - Roethlein, Christoph A1 - Zhang, Gong A1 - Techen, Anne A1 - Dosche, Carsten A1 - Ignatova, Zoya T1 - Natural osmolytes remodel the aggregation pathway of mutant huntingtin exon 1 JF - Biochemistry N2 - In response to stress small organic compounds termed osmolytes are ubiquitously accumulated in all cell types to regulate the intracellular solvent quality and to counteract the deleterious effect on the stability and function of cellular proteins. Given the evidence that destabilization of the native state of a protein either by mutation or by environmental changes triggers the aggregation in the neurodegenerative pathologies, the modulation of the intracellular solute composition with osmolytes is an attractive strategy to stabilize an aggregating protein. Here we report the effect of three natural osmolytes on the in vivo and in vitro aggregation landscape of huntingtin exon 1 implicated in the Huntington's disease. Trimethylamine N-oxide (TMAO) and proline redirect amyloid fibrillogenesis of the pathological huntingtin exon 1 to nonamyloidogenic amorphous assemblies via two dissimilar molecular mechanisms. TMAO causes a rapid formation of bulky amorphous aggregates with minimally exposed surface area, whereas proline solubilizes the monomer and suppresses the accumulation of early transient aggregates. Conversely, glycine betaine enhances fibrillization in a fashion reminiscent of the genesis of functional amyloids. Strikingly, none of the natural osmolytes can completely abrogate the aggregate formation; however, they redirect the amyloidogenesis into alternative, nontoxic aggregate species. Our study reveals new insights into the complex interactions of osmoprotectants with polyQaggregates. Y1 - 2011 U6 - https://doi.org/10.1021/bi1018368 SN - 0006-2960 VL - 50 IS - 12 SP - 2048 EP - 2060 PB - American Chemical Society CY - Washington ER - TY - JOUR A1 - Lahn, Mattes A1 - Dosche, Carsten A1 - Hille, Carsten T1 - Two-photon microscopy and fluorescence lifetime imaging reveal stimulus-induced intracellular Na+ and Cl- changes in cockroach salivary acinar cells JF - American journal of physiology : Cell physiology N2 - Lahn M, Dosche C, Hille C. Two-photon microscopy and fluorescence lifetime imaging reveal stimulus-induced intracellular Na+ and Cl- changes in cockroach salivary acinar cells. Am J Physiol Cell Physiol 300: C1323-C1336, 2011. First published February 23, 2011; doi: 10.1152/ajpcell.00320.2010.-The intracellular ion homeostasis in cockroach salivary acinar cells during salivation is not satisfactorily understood. This is mainly due to technical problems regarding strong tissue autofluorescence and ineffective ion concentration quantification. For minimizing these problems, we describe the successful application of two-photon (2P) microscopy partly in combination with fluorescence lifetime imaging microscopy (FLIM) to record intracellular Na+ and Cl- concentrations ([Na+](i), [Cl-](i)) in cockroach salivary acinar cells. Quantitative 2P-FLIM Cl- measurements with the dye N-(ethoxycarbonylmethyl)-6-methoxy-quinolinium bromide indicate that the resting [Cl-](i) is 1.6 times above the Cl- electrochemical equilibrium but is not influenced by pharmacological inhibition of the Na+-K+-2Cl(-) cotransporter (NKCC) and anion exchanger using bumetanide and 4,4'-diisothiocyanatodihydrostilbene-2,2'-disulfonic acid disodium salt. In contrast, rapid Cl- reuptake after extracellular Cl- removal is almost totally NKCC mediated both in the absence and presence of dopamine. However, in physiological saline [Cl-](i) does not change during dopamine stimulation although dopamine stimulates fluid secretion in these glands. On the other hand, dopamine causes a decrease in the sodium-binding benzofuran isophthalate tetra-ammonium salt (SBFI) fluorescence and an increase in the Sodium Green fluorescence after 2P excitation. This opposite behavior of both dyes suggests a dopamine-induced [Na+](i) rise in the acinar cells, which is supported by the determined 2P-action cross sections of SBFI. The [Na+](i) rise is Cl- dependent and inhibited by bumetanide. The Ca2+-ionophore ionomycin also causes a bumetanide-sensitive [Na+](i) rise. We propose that a Ca2+-mediated NKCC activity in acinar peripheral cells attributable to dopamine stimulation serves for basolateral Na+ uptake during saliva secretion and that the concomitantly transported Cl- is recycled back to the bath. KW - epithelial ion transport KW - Na+-K+-2Cl(-) cotransporter KW - MQAE KW - SBFI KW - 2P cross section Y1 - 2011 U6 - https://doi.org/10.1152/ajpcell.00320.2010 SN - 0363-6143 VL - 300 IS - 6 SP - C1323 EP - C1336 PB - American Chemical Society CY - Bethesda ER - TY - JOUR A1 - Mutig, Kerim A1 - Kahl, Thomas A1 - Saritas, Turgay A1 - Godes, Michael A1 - Persson, Pontus A1 - Bates, James A1 - Raffi, Hajamohideen A1 - Rampoldi, Luca A1 - Uchida, Shinichi A1 - Hille, Carsten A1 - Dosche, Carsten A1 - Kumar, Satish A1 - Castaneda-Bueno, Maria A1 - Gamba, Gerardo A1 - Bachmann, Sebastian T1 - Activation of the Bumetanide-sensitive Na+, K+,2Cl(-) Cotransporter (NKCC2) Is Facilitated by Tamm-Horsfall Protein in a Chloride-sensitive Manner JF - The journal of biological chemistry N2 - Active transport of NaCl across thick ascending limb (TAL) epithelium is accomplished by Na+, K+,2Cl(-) cotransporter (NKCC2). The activity of NKCC2 is determined by vasopressin (AVP) or intracellular chloride concentration and includes its amino-terminal phosphorylation. Co-expressed Tamm-Horsfall protein (THP) has been proposed to interact with NKCC2. We hypothesized that THP modulates NKCC2 activity in TAL. THP-deficient mice (THP-/-) showed an increased abundance of intracellular NKCC2 located in subapical vesicles (+47% compared with wild type (WT) mice), whereas base-line phosphorylation of NKCC2 was significantly decreased (-49% compared with WT mice), suggesting reduced activity of the transporter in the absence of THP. Cultured TAL cells with low endogenous THP levels and low base-line phosphorylation of NKCC2 displayed sharp increases in NKCC2 phosphorylation (+38%) along with a significant change of intracellular chloride concentration upon transfection with THP. In NKCC2-expressing frog oocytes, co-injection with THP cRNA significantly enhanced the activation of NKCC2 under low chloride hypotonic stress (+112% versus +235%). Short term (30 min) stimulation of the vasopressin V2 receptor pathway by V2 receptor agonist (deamino-cis-D-Arg vasopressin) resulted in enhanced NKCC2 phosphorylation in WT mice and cultured TAL cells transfected with THP, whereas in the absence of THP, NKCC2 phosphorylation upon deamino-cis-D-Arg vasopressin was blunted in both systems. Attenuated effects of furosemide along with functional and structural adaptation of the distal convoluted tubule in THP-/- mice supported the notion that NaCl reabsorption was impaired in TAL lacking THP. In summary, these results are compatible with a permissive role for THP in the modulation of NKCC2-dependent TAL salt reabsorptive function. Y1 - 2011 U6 - https://doi.org/10.1074/jbc.M111.222968 SN - 0021-9258 VL - 286 IS - 34 SP - 30200 EP - 30210 PB - American Society for Biochemistry and Molecular Biology CY - Bethesda ER - TY - JOUR A1 - Riebe, Daniel A1 - Zühlke, Martin A1 - Zenichowski, Karl A1 - Beitz, Toralf A1 - Dosche, Carsten A1 - Löhmannsröben, Hans-Gerd T1 - Characterization of rhodamine 6G release in electrospray ionization by means of spatially resolved fluorescence spectroscopy JF - Zeitschrift für physikalische Chemie : international journal of research in physical chemistry and chemical physics N2 - In the present work, the density distribution of rhodamine 6G ions (R6G) in the gas phase and the droplets of an electrospray plume was studied by spatial and spectral imaging. The intention is to contribute to the fundamental understanding of the release mechanism of gaseous R6G in the electrospray ionization (ESI) process. Furthermore, the influence of ESI-parameters on the release efficiency of R6G, e. g. solvent flow, R6G and salt concentration were examined via direct fluorescence imaging of R6G. A solvent-shift of the fluorescence maximum,lambda(max) = 555 nm in methanolic solution and lambda(max) = 505 nm in gas phase, allows the discrimination between solvated and gaseous R6G. Two experimental setups were used for our measurements. In the first experiment, the R6G fluorescence and the light scattered from the spray plume were imaged in two spatial dimensions using a tunable wavelength filter. The second experiment was designed for obtaining 1-dimensional spatially resolved emission spectra of the spray. Here, the intensity distribution of solvated and gaseous R6G as well as scattered light (lambda = 355 nm) were measured simultaneously. The results show the distribution of gaseous R6G in the plane, orthogonal to the ESI capillary, decreasing slightly towards the spray center and showing maxima at the cone margins. The distribution of gaseous R6G confirms the preferred release of gaseous ions from nano-droplets, indicating the ion evaporation model (IEM) to be the dominating release mechanism. Up to now, only a few fluorescence spectra of ionic compounds in the gas phase were published because the measurement of emission spectra of mass-selected ions in an ion trap is experimentally challenging. The fluorescence spectrum of gaseous lucigenin at atmospheric pressure is reported for the first time. This spectrum of lucigenin in the gas phase exhibits a blue shift of about Delta lambda = 10 nm in comparison to the corresponding spectrum in methanol. KW - Fluorescence KW - Electrospray Ionization KW - Rhodamine 6G KW - Gaseous Ions KW - Lucigenin Y1 - 2011 U6 - https://doi.org/10.1524/zpch.2011.0149 SN - 0942-9352 VL - 225 IS - 9-10 SP - 1055 EP - 1072 PB - De Gruyter Oldenbourg CY - München ER - TY - JOUR A1 - Kammer, Stefan A1 - Starke, Ines A1 - Pietrucha, Andreas A1 - Kelling, Alexandra A1 - Mickler, Wulfhard A1 - Schilde, Uwe A1 - Dosche, Carsten A1 - Kleinpeter, Erich A1 - Holdt, Hans-Jürgen T1 - 1,12-Diazaperylene and 2,11-dialkylated-1,12-diazaperylene iridium(III) complexes [Ir((CN)-N-boolean AND)(2)((NN)-N-boolean AND)]PF6: new supramolecular assemblies JF - Dalton transactions : a journal of inorganic chemistry, including bioinorganic, organometallic, and solid-state chemistry N2 - A series of new monocationic iridium(III) complexes [Ir((CN)-N-boolean AND)(2)((NN)-N-boolean AND)]PF6 with "large-surface" alpha,alpha'-diimin ligands (NN)-N-boolean AND (dap = 1,12-diazaperylene, dmedap = 2,11-dimethyl-1,12-diazaperylene, dipdap = 2,11-diisopropyl-1,12-diazaperylene) and different cyclometalating ligands (CN)-N-boolean AND (piq = 1-phenylisoquinoline, bzq = benzo[h]quinoline, ppz = 1-phenylpyrazole, thpy = 2-(2-thienyl)pyridine, ppy = 2-phenylpyridine, meppy = 2-(4-methylphenyl)pyridine, dfppy = 2-(2,4-difluorophenyl)pyridine) were synthesized. The solid structures of the complexes [Ir(piq)(2)(dap)]PF6, [Ir(bzq)(2)(dap)]PF6, [Ir(ppy)(2)(dipdap)]PF6, [Ir(piq)(2)(dmedap)]PF6, [Ir(ppy)(2)(dap)]PF6 and [Ir(ppz)(2)(dap)]PF6 are reported. In [Ir(piq)(2)(dap)]PF6, the dap ligand and one of the piq ligands of each cationic complex are involved in pi-pi stacking interactions forming supramolecular channels running along the crystallographic c axis. In the crystalline [Ir(bzq)(2)(dap)]PF6 pi-pi stacking interactions between the metal complexes lead to the formation of a 2D layer structure. In addition, CH-pi interactions were found in all compounds, which are what stabilizes the solid structure. In particular, a significant number of them were found in [Ir(piq)(2)(dap)]PF6 and [Ir(bzq)(2)(dap)]PF6. The crystal structures of [Ir(ppy)(2)(dipdap)]PF6 and [Ir(ppy)(2)(dmedap)]PF6 are also presented, being the first examples of bis-cyclometalated iridium(III) complexes with phenanthroline-type alpha,alpha'-diimin ligands bearing bulky alkyl groups in the neighbourhood of the N-donor atoms. These ligands implicate a distorted octahedral coordination geometry that in turn destabilized the Ir-N-N boolean AND N bonds. The new iridium (III) complexes are not luminescent. All compounds show an electrochemically irreversible anodic peak between 1.15 and 1.58 V, which is influenced by the different cyclometalated ligands. All of the new complexes show two reversible successive one-electron "large-surface" ligand-centred reductions around -0.70 V and -1.30 V. Electrospray ionisation mass spectrometry (ESI-MS) and collision induced decomposition (CID) measurements were used to investigate the stability of the new complexes. Thereby, the stability agreed well with the order of the Ir-N-N boolean AND N bond lengths. Y1 - 2012 U6 - https://doi.org/10.1039/c2dt30412k SN - 1477-9226 VL - 41 IS - 34 SP - 10219 EP - 10227 PB - Royal Society of Chemistry CY - Cambridge ER - TY - JOUR A1 - Chowdhury, Mita Mullick A1 - Dosche, Carsten A1 - Loehmannsröben, Hans-Gerd A1 - Leimkühler, Silke T1 - Dual role of the molybdenum cofactor biosynthesis protein MOCS3 in tRNA thiolation and molybdenum cofactor biosynthesis in humans JF - The journal of biological chemistry N2 - We studied two pathways that involve the transfer of persulfide sulfur in humans, molybdenum cofactor biosynthesis and tRNA thiolation. Investigations using human cells showed that the two-domain protein MOCS3 is shared between both pathways. MOCS3 has an N-terminal adenylation domain and a C-terminal rhodanese-like domain. We showed that MOCS3 activates both MOCS2A and URM1 by adenylation and a subsequent sulfur transfer step for the formation of the thiocarboxylate group at the C terminus of each protein. MOCS2A and URM1 are beta-grasp fold proteins that contain a highly conserved C-terminal double glycine motif. The role of the terminal glycine of MOCS2A and URM1 was examined for the interaction and the cellular localization with MOCS3. Deletion of the C-terminal glycine of either MOCS2A or URM1 resulted in a loss of interaction with MOCS3. Enhanced cyan fluorescent protein and enhanced yellow fluorescent protein fusions of the proteins were constructed, and the fluorescence resonance energy transfer efficiency was determined by the decrease in the donor lifetime. The cellular localization results showed that extension of the C terminus with an additional glycine of MOCS2A and URM1 altered the localization of MOCS3 from the cytosol to the nucleus. Y1 - 2012 U6 - https://doi.org/10.1074/jbc.M112.351429 SN - 0021-9258 VL - 287 IS - 21 SP - 17297 EP - 17307 PB - American Society for Biochemistry and Molecular Biology CY - Bethesda ER - TY - JOUR A1 - Techen, Anne A1 - Hille, Carsten A1 - Dosche, Carsten A1 - Kumke, Michael Uwe T1 - Fluorescence study of drug-carrier interactions in CTAB/PBS buffer model systems JF - Journal of colloid and interface science N2 - The well-known cationic surfactant hexadecyltrimethylammonium bromide (CTAB) was used as a model carrier to study drug-carrier interactions with fluorescence probes (5-hexadecanoylaminofluorescein (HAF) and 2,10-bis-(3-aminopropyloxy)dibenzo[aj]perylene-8,16-dione (NIR 628) by applying ensemble as well as single molecule fluorescence techniques. The impact of the probes on the micelle parameters (critical micelle concentration, average aggregation number, hydrodynamic radius) was investigated under physiological conditions. In the presence of additional electrolytes, such as buffer, the critical micelle concentration decreased by a factor of about 10. In contrast, no influence of the probes on the critical micelle concentration and on average aggregation number was observed. The results show that HAF does not affect the characteristics of CTAB micelles. Analyzing fluorescence correlation spectroscopy data and time-resolved anisotropy decays in terms of the "two-step" in combination with the "wobbling-in-cone" model, it was proven that HAF and NIR 628 are differently associated with the micelles. Based on ensemble and single molecule fluorescence experiments, intra- and intermicellar energy transfer process between the two dyes were probed and characterized. KW - Hexadecyltrimethylammonium bromide KW - 5-Hexadecanoylaminofluorescein KW - 2,10-Bis-(3-aminopropyloxy)dibenzo[aj]perylene-8,16-dione KW - Fluorescence correlation spectroscopy KW - Fluorescence anisotropy KW - Single-molecule FRET Y1 - 2012 U6 - https://doi.org/10.1016/j.jcis.2012.03.063 SN - 0021-9797 VL - 377 SP - 251 EP - 261 PB - Elsevier CY - San Diego ER -