Seebeck, Peltier, and Soret effects: On different formalisms for transport equations in thermogalvanic cells.

Thermogalvanic cells convert waste heat directly to electric work. There is an abundance of waste heat in the world and thermogalvanic cells may be underused. We discuss theoretical tools that can help us understand and therefore improve on cell performance. One theory is able to describe all aspects of the energy conversion: nonequilibrium thermodynamics. We recommend to use the theory with operationally defined, independent variables, as others have done before. These describe well-defined experiments. Three invariance criteria serve as a basis for any description: of local electroneutrality, entropy production invariance, and emf's independence of the frame of reference. Alternative formalisms, using different sets of variables, start with ionic or neutral components. We show that the heat flux is not the same in the two formalisms and derive a new relationship between the heat fluxes. The heat flux enters the definition of the Peltier coefficient and is essential for the understanding of the Peltier heat at the electrode interfaces and of the Seebeck coefficient of the cell. The Soret effect can occur independently of any Seebeck effect, but the Seebeck effect will be affected by the presence of a Soret effect. Common misunderstandings are pointed out. Peltier coefficients are needed for the interpretation and design of experiments.

Legendre-Fenchel transforms capture layering transitions in porous media.

We have investigated the state of a nanoconfined fluid in a slit pore in the canonical and isobaric ensembles. The systems were simulated with molecular dynamics simulations. The fluid has a transition to a close-packed structure when the height of the slit approaches the particle diameter. The Helmholtz energy is a non-convex function of the slit height if the number of particles does not exceed that of one monolayer. As a consequence, the Legendre transform cannot be applied to obtain the Gibbs energy. The Gibbs energy of a non-deformable slit pore can be transformed into the Helmholtz energy of a deformable slit pore using the Legendre-Fenchel transform. The Legendre-Fenchel transform corresponds to the Maxwell construction of equal areas.

Perspectives on Thermoelectric Energy Conversion in Ion-Exchange Membranes.

By thermoelectric power generation we mean the creation of electrical power directly from a temperature gradient. Semiconductors have been mainly used for this purpose, but these imply the use of rare and expensive materials. We show in this review that ion-exchange membranes may be interesting alternatives for thermoelectric energy conversion, giving Seebeck coefficients around 1 mV/K. Laboratory cells with Ag|AgCl electrodes can be used to find the transported entropies of the ions in the membrane without making assumptions. Non-equilibrium thermodynamics can be used to compute the Seebeck coefficient of this and other cells, in particular the popular cell with calomel electrodes. We review experimental results in the literature on cells with ion-exchange membranes, document the relatively large Seebeck coefficient, and explain with the help of theory its variation with electrode materials and electrolyte concentration and composition. The impact of the membrane heterogeneity and water content on the transported entropies is documented, and it is concluded that this and other properties should be further investigated, to better understand how all transport properties can serve the purpose of thermoelectric energy conversion.

Fine-tuned ATP signals are acute mediators in osteocyte mechanotransduction.

Osteocytes are considered the primary mechanosensors of bone, but the signaling pathways they apply in mechanotransduction are still incompletely investigated and characterized. A growing body of data strongly indicates that P2 receptor signaling among osteoblasts and osteoclasts has regulatory effects on bone remodeling. Therefore, we hypothesized that ATP signaling is also applied by osteocytes in mechanotransduction. We applied a short fluid pulse on MLO-Y4 osteocyte-like cells during real-time detection of ATP and demonstrated that mechanical stimulation activates the acute release of ATP and that these acute ATP signals are fine-tuned according to the magnitude of loading. ATP release was then challenged by pharmacological inhibitors, which indicated a vesicular release pathway for acute ATP signals. Finally, we showed that osteocytes express functional P2X2 and P2X7 receptors and respond to even low concentrations of nucleotides by increasing intracellular calcium concentration. These results indicate that in osteocytes, vesicular ATP release is an acute mediator of mechanical signals and the magnitude of loading. These and previous results, therefore, implicate purinergic signaling as an early signaling pathway in osteocyte mechanotransduction.

In vivo experimental stroke and in vitro organ culture induce similar changes in vasoconstrictor receptors and intracellular calcium handling in rat cerebral arteries.

Cerebral arteries subjected to different types of experimental stroke upregulate their expression of certain G-protein-coupled vasoconstrictor receptors, a phenomenon that worsens the ischemic brain damage. Upregulation of contractile endothelin B (ET(B)) and 5-hydroxytryptamine 1B (5-HT(1B)) receptors has been demonstrated after subarachnoid hemorrhage and global ischemic stroke, but the situation is less clear after focal ischemic stroke. Changes in smooth muscle calcium handling have been implicated in different vascular diseases but have not hitherto been investigated in cerebral arteries after stroke. Here, we evaluate changes of ET(B) and 5-HT(1B) receptors, intracellular calcium levels, and calcium channel expression in rat middle cerebral artery (MCA) after focal cerebral ischemia and in vitro organ culture, a proposed model of vasoconstrictor receptor changes after stroke. Rats were subjected to 2 h MCA occlusion followed by reperfusion for 1 or 24 h. Alternatively, MCAs from naïve rats were cultured for 1 or 24 h. ET(B) and 5-HT(1B) receptor-mediated contractions were evaluated by wire myography. Receptor and channel expressions were measured by real-time PCR and immunohistochemistry. Intracellular calcium was measured by FURA-2. Expression and contractile functions of ET(B) and 5-HT(1B) receptors were strongly upregulated and slightly downregulated, respectively, 24 h after experimental stroke or organ culture. ET(B) receptor-mediated contraction was mediated by calcium from intracellular and extracellular sources, whereas 5-HT(1B) receptor-mediated contraction was solely dependent on extracellular calcium. Organ culture and stroke increased basal intracellular calcium levels in MCA smooth muscle cells and decreased the expression of inositol triphosphate receptor and transient receptor potential canonical calcium channels, but not voltage-operated calcium channels.

DoloTest in General Practice Study: Sensitivity and Specificity Screening for Depression.

Background. Coexistence of pain and depression has significant impact on the patient's quality of life and treatment outcome. DoloTest is a pain and HRQoL assessment tool developed to provide shared understanding between the clinician and the patient of the condition by a visual profile. Aim. To find the sensitivity and specificity of DoloTest as a screening tool for depression for patients in primary care. Methods. All patients coming to a primary care clinic were asked to fill in a DoloTest and a Major Depression Inventory. Results. 715 (68.5%) of 1044 patients entered the study. 34.4% came due to pain. 16.1% met depression criteria, and 26.8% of patients coming due to pain met criteria for depression. 65.6% of the men and 54.2% of the women meeting the criteria for depression came due to pain. Depressed patients had statistically significant higher scores on all DoloTest domains. Selecting the cutoff value for the domain "low spirits" to be "65" (0-100) for depression gave a sensitivity of 78% (70-85%) and a specificity of 95% (93-96%) for meeting depression criteria. Conclusion. DoloTest can with a high sensitivity and specificity identify persons meeting criteria for depression and is an easy-to-use screening tool to identify patients with the coexistence of pain and depression.

Cellular and subcellular localization of flavin-monooxygenases involved in glucosinolate biosynthesis.

Glucosinolates are amino acid-derived secondary metabolites with diverse biological activities dependent on chemical modifications of the side chain. Five flavin-monooxygenases FMO(GS-OX1-5) have recently been identified as aliphatic glucosinolate side chain modification enzymes in Arabidopsis thaliana that catalyse the generation of methylsulphinylalkyl glucosinolates, which can be hydrolysed to products with distinctive benefits for human health and plant defence. Though the localization of most aliphatic glucosinolate biosynthetic enzymes has been determined, little is known about where the side chain modifications take place despite their importance. Hence, the spatial expression pattern of FMO(GS-OX1-5) genes in Arabidopsis was investigated by expressing green fluorescent protein (GFP) and ß-glucuronidase (GUS) fusion genes controlled by FMO(GS-OX1-5) promoters. The cellular compartmentation of FMO(GS-OX1) was also detected by transiently expressing a FMO(GS-OX1)-yellow fluorescent protein (YFP) fusion protein in tobacco leaves. The results showed that FMO(GS-OX1-5) were expressed basically in vascular tissues, especially in phloem cells, like other glucosinolate biosynthetic genes. They were also found in endodermis-like cells in flower stalk and epidermal cells in leaf, which is a location that has not been reported for other glucosinolate biosynthetic genes. It is suggested that the spatial expression pattern of FMO(GS-OX1-5) determines the access of enzymes to their substrate and therefore affects the glucosinolate profile. FMO(GS-OX1)-YFP fusion protein analysis identified FMO(GS-OX1) as a cytosolic protein. Together with the subcellular locations of the other biosynthetic enzymes, an integrated map of the multicompartmentalized aliphatic glucosinolate biosynthetic pathway is discussed.

Neurogenic inflammation: a study of rat trigeminal ganglion.

Calcitonin gene-related peptide (CGRP) is linked to neurogenic inflammation and to migraine. Activation of the trigeminovascular system plays a prominent role during migraine attacks with the release of CGRP. The trigeminal ganglion (TG) contains three main cell types: neurons, satellite glial cells (SGC) and Schwann cells; the first two have before been studied in vitro separately. Culture of rat TG provides a method to induce inflammation and the possibility to evaluate the different cell types in the TG simultaneously. We investigated expression levels of various inflammatory cytokines on mRNA level using RT-PCR arrays and qRT-PCR; furthermore expression at protein level was studied using immunohistochemistry. We report that (1) organ culture of the TG is possible with preserved morphology, (2) organ culture is associated with enhanced expression of cytokines and mitogen-activated protein kinases (MAPKs) primarily in neurons, (3) CGRP can induce expression of some cytokines and (4) cytokine expression is still upregulated following MAPK pathway inhibition by MEK inhibitor U0126 and pp38 inhibitor SB202192, but the cytokine expression is abolished when co-incubating with the JNK inhibitor SP600125. This method may be of value to examine local TG inflammation, putatively involved in the pathophysiology of some forms of primary headaches.

LPS from Porphyromonas gingivalis increases the sensitivity of contractile response mediated by endothelin-B (ET(B)) receptors in cultured endothelium-intact rat coronary arteries.

The purpose of our study was to examine if lipopolysaccharide (LPS) from Porphyromonas gingivalis (P.g.) modifies the vasomotor responses to Endothelin-1 (ET-1) and Sarafotoxin 6c (S6c) in rat coronary arteries. The arteries were studied directly or following organ culture for 24 h in absence and presence of 2.5EU/ml LPS. The contractile responses of coronary arteries were investigated by using the selective ETB receptor agonist S6c (1 pM-0.3 µM) and ET-1 (1 pM-0.3 µM). The functional studies demonstrated an augmented contractile response only to S6c in isolated rat coronary arteries after organ culture (with or without LPS). These contractile responses by S6c were blocked by the selective ETB receptor antagonist BQ788 in both vessel groups. The augmented contractile response to S6c was supported by immunohistochemistry, where a significant increase in fluorescence intensity for ETB receptors in smooth muscle cells was observed after organ culture. The presence of LPS in the culture medium significantly increased the sensitivity of endothelium-intact coronary artery to S6c as compared to endothelium-denuded segments. Our results showed a significant increase in both ETB receptor protein levels and S6c-induced maximal contraction in coronary arteries upon 24 h of organ culture, which was further sensitized by LPS.

Introduction and Validation of DoloTest(®): a new health-related quality of life tool used in pain patients.

PURPOSE: The aim of the publication is to introduce DoloTest(®) and to document the validation of DoloTest(®) . DoloTest(®) is a health-related quality of life (HRQoL) tool for pain patients with low-time burden involving the patient in interpretation of the test result by creating a graphic presentation of the test result in a DoloTest(®) Profile and therefore, suitable use in clinical setting. METHODS: Validation against SF-36, 246 participants in four pain clinics. MAIN OUTCOME MEASURES: Face validity (domains) cognitive interviewing (time and patient's perception), factor analysis (factors and eigenvalues), construct validity (correlation coefficients), Reliability: Intern consistency (Cronbach's alpha), test-retest stability (correlation coefficients). RESULTS: Face validation: the eight domains most important for assessment of the patients with persistent pain were selected. The factor analysis shows equal relevance and weighting of all domains. Construct validation against SF-36™ shows positive correlation within each domain; correlation coefficients were between 0.47 and 0.69. Reliability was tested with Cronbach's alpha and test-retest. Cronbach's alpha values were 0.615 to 0.715. Correlation coefficients for linear regressions of test and retest for the DoloTest(®) and SF-36 were all positive. INTRODUCTION AND VALIDATION OF DOLOTEST(®): A new HRQoL tool used in pain patients.

Monitoring reactive oxygen species formation and localisation in living cells by use of the fluorescent probe CM-H(2)DCFDA and confocal laser microscopy.

Reactive oxygen species (ROS) develop as a consequence of wounding, light stress and chemical imbalances but act also as signals in living cells. The integrity of cells is seriously endangered, if ROS cannot be controlled by scavenging molecules and other repair mechanisms of the cell. For studying ROS development and signalling under stress, a reliable indicator is needed. We have tested the ROS sensitive dye 5-(and-6) chloromethyl-2',7' dichlorodihydrofluorescein diacetate acetyl ester (CM-H(2)DCFDA) using onion bulb scale and leaf epidermis as well as Arabidopsis leaves and protoplasts. ROS were generated by several fundamentally different methods-externally applied hydrogen peroxide, heat shock, high light or wounding. Confocal microscopy and fluorescence quantification over time showed that the indicator responds in an additive and dose-dependent manner. The response to externally applied hydrogen peroxide followed saturation kinetics, consistent with a channel-mediated uptake of the stressor across the plasma membrane. An inherent problem of the tested indicator was the uneven uptake in tissues, as compared with protoplasts, making it difficult to discriminate an uneven indicator distribution from an uneven ROS distribution. However, in protoplasts and under carefully designed preparation conditions CM-H(2)DCFDA is a useful general ROS indicator. Subcellularly, the de-esterified probe localised to the cytosol, to mitochondria and to chloroplasts.

Non-invasive method for in vivo detection of chlorophyll precursors.

Traditionally chlorophyll (Chl) and Chl precursors have been studied in vitro or in leaf tissue at low temperature. These methods are destructive and make it impossible to work with the same individual plant later on. In this paper we present a method for in vivo detection of Chl and its precursors in seedling plants which can be rescued for further studies. Multi-photon microscopy, which we show to be more reliable in vivo than UV-laser microscopy, is used to detect precursors in the biosynthetic pathway leading to Chl. The sensitivity and ability to distinguish different precursors with this system is compared to current methods. Furthermore, we report on optimization of the spectral scanning method with the aim to minimize the excitation light-evoked photo-conversion of the chlorophyll precursors.

Arabidopsis protein kinase PKS5 inhibits the plasma membrane H+ -ATPase by preventing interaction with 14-3-3 protein.

Regulation of the trans-plasma membrane pH gradient is an important part of plant responses to several hormonal and environmental cues, including auxin, blue light, and fungal elicitors. However, little is known about the signaling components that mediate this regulation. Here, we report that an Arabidopsis thaliana Ser/Thr protein kinase, PKS5, is a negative regulator of the plasma membrane proton pump (PM H+ -ATPase). Loss-of-function pks5 mutant plants are more tolerant of high external pH due to extrusion of protons to the extracellular space. PKS5 phosphorylates the PM H+ -ATPase AHA2 at a novel site, Ser-931, in the C-terminal regulatory domain. Phosphorylation at this site inhibits interaction between the PM H+ -ATPase and an activating 14-3-3 protein in a yeast expression system. We show that PKS5 interacts with the calcium binding protein SCaBP1 and that high external pH can trigger an increase in the concentration of cytosolic-free calcium. These results suggest that PKS5 is part of a calcium-signaling pathway mediating PM H+ -ATPase regulation.

Specific aquaporins facilitate the diffusion of hydrogen peroxide across membranes.

The metabolism of aerobic organisms continuously produces reactive oxygen species. Although potentially toxic, these compounds also function in signaling. One important feature of signaling compounds is their ability to move between different compartments, e.g. to cross membranes. Here we present evidence that aquaporins can channel hydrogen peroxide (H2O2). Twenty-four aquaporins from plants and mammals were screened in five yeast strains differing in sensitivity toward oxidative stress. Expression of human AQP8 and plant Arabidopsis TIP1;1 and TIP1;2 in yeast decreased growth and survival in the presence of H2O2. Further evidence for aquaporin-mediated H2O2 diffusion was obtained by a fluorescence assay with intact yeast cells using an intracellular reactive oxygen species-sensitive fluorescent dye. Application of silver ions (Ag+), which block aquaporin-mediated water diffusion in a fast kinetics swelling assay, also reversed both the aquaporin-dependent growth repression and the H2O2-induced fluorescence. Our results present the first molecular genetic evidence for the diffusion of H2O2 through specific members of the aquaporin family.

Molecular identification of mycorrhizal fungi in Neuwiedia veratrifolia (Orchidaceae).

We here apply a previously described method for identification of single peloton orchid mycorrhiza to a key orchid group and extend the usefulness in the heterobasidiomycetes of an existing fungal database for identification of mycorrhizal fungi. We amplified and sequenced mitochondrial ribosomal large subunit DNA from fungi in roots of Neuwiedia veratrifolia (Orchidaceae), a member of the small subfamily Apostasioideae that is sister to the remainder of Orchidaceae, and used the extended database to identify the mycorrhizal fungi. Sequences from fungi cultured from Neuwiedia roots and from direct peloton amplifications were analyzed cladistically with sequences determined from reference fungal collections and published sequences. The fungi from Neuwiedia are referred to the heterobasidiomycetous orders Tulasnellales and Ceratobasidiales, indicating that apostasioids utilize the same fungi as other photosynthetic orchids. The majority of Neuwiedia mycobionts came together in a clade with Tulasnella species, but some were most closely related to Thanatephorus. In some cases members of these two clades were isolated from the same orchid plant, providing another example of multiple mycobionts occurring in a single plant.