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A regional geoid model for the Argentine part of the Isla Grande de Tierra del Fuego, established in previous works on the basis of GPS levelling, suffers a lack of observation data in the remote south-western investigation area. In order to improve the data distribution in this region, the mean lake level of Lago Fagnano has been regarded as a natural indicator for the local geoid. Using a GPS buoy and pressure tide gauges, a method to determine the mean lake surface topography with respect to the ellipsoid has been developed. It is shown that the obtained lake level geometry is essentially controlled by the regional gravity field. The derived information on the mean lake level has been included in the geoid model, which results in a more detailed and plausible representation of the regional geoid.

Introduction: Head and neck squamous cell carcinoma (HNSCC) patients suffer from frequent local recurrences that negatively impact on prognosis. Hence, distinguishing tumor and normal tissue is of clinical importance as it may improve the detection of residual tumor tissue in surgical resection margins and during imaging-based surgery planning. Differences in O 2 consumption (OC) can be used to this aim, as they provide options for improved surgical, image-guided approaches., Methods: In the present study, the potential of a fluorescent sensor foil-based technology to quantify OC in HNSCC was evaluated in an in vitro 3D model and in situ in patients., Results: In vitro measurements of OC using hypopharyngeal and esophageal cell lines allowed a specific detection of tumor cell spheroids embedded together with cancer-associated fibroblasts in type I collagen extracellular matrix down to a diameter of 440 µm. Pre-surgery in situ measurements were conducted with a handheld recording device and sensor foils with an oxygen permeable membrane and immobilized O 2 -reactive fluorescent dyes. Lateral tongue carcinoma and carcinoma of the floor of the mouth were chosen for analysis owing to their facilitated accessibility. OC was evaluated over a time span of 60 seconds and was significantly higher in tumor tissue compared to healthy mucosa in the vicinity of the tumor., Discussion: Hence, OC quantification using fluorescent sensor foil-based technology is a relevant parameter for the differentiation of tumor tissue of the head and neck region and may support surgery planning., Competing Interests: Co-authors RM and GL are employees of PreSens Precision Sensing GmbH, Regensburg, Germany. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Stocker, Blancke Soares, Liebsch, Meier, Canis, Gires and Haubner.)

After the development of 3D cell culture methods in the middle of the last century and the plethora of data generated with this culture configuration up to date, it could be shown that a three-dimensional arrangement of cells in most of the cases leads to a more physiological behavior of the generated tissue. However, a major determinant for an organotypic function, namely, the dissolved oxygen concentration in the used in vitro -system, has been neglected in most of the studies. This is due to the fact that the oxygen measurement in the beginning was simply not feasible and, if so, disturbed the measurement and/or the in vitro -system itself. This is especially true for the meanwhile more widespread use of 3D culture systems. Therefore, the tissues analyzed by these techniques can be considered as the Schrödinger's cat in 3D cell biology. In this perspective paper we will outline how the measurement and, moreover, the regulation of the dissolved oxygen concentration in vitro -3D culture systems could be established at all and how it may be possible to determine the oxygen concentration in organoid cultures and the respiratory capacity via mito stress tests, especially in spheroids in the size range of a few hundred micrometers, under physiological culture conditions, without disturbances or stress induction in the system and in a high-throughput fashion. By this, such systems will help to more efficiently translate tissue engineering approaches into new in vitro -platforms for fundamental and applied research as well as preclinical safety testing and clinical applications., Competing Interests: Author GL is employed by the company PreSens Precision Sensing GmbH. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Gottwald, Grün, Nies and Liebsch.)

Oxygen concentration plays a crucial role in (3D) cell culture. However, the oxygen content in vitro is usually not comparable to the in vivo situation, which is partly due to the fact that most experiments are performed under ambient atmosphere supplemented with 5% CO 2 , which can lead to hyperoxia. Cultivation under physiological conditions is necessary, but also fails to have suitable measurement methods, especially in 3D cell culture. Current oxygen measurement methods rely on global oxygen measurements (dish or well) and can only be performed in 2D cultures. In this paper, we describe a system that allows the determination of oxygen in 3D cell culture, especially in the microenvironment of single spheroids/organoids. For this purpose, microthermoforming was used to generate microcavity arrays from oxygen-sensitive polymer films. In these oxygen-sensitive microcavity arrays (sensor arrays), spheroids cannot only be generated but also cultivated further. In initial experiments we could show that the system is able to perform mitochondrial stress tests in spheroid cultures to characterize mitochondrial respiration in 3D. Thus, with the help of sensor arrays, it is possible to determine oxygen label-free and in real-time in the immediate microenvironment of spheroid cultures for the first time., Competing Interests: Author GL is employed by PreSens Precision Sensing GmbH. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Grün, Pfeifer, Liebsch and Gottwald.)

Background: pO 2 and pH are physiological parameters relevant for different processes in health and disease, including wound healing and cancer progression. Head and neck squamous cell carcinomas (HNSCC) and oesophageal squamous cell carcinomas (ESCC) have a high rate of local recurrence that is partly related to treatment-resistant residual tumour cells. Hence, novel diagnostic tools are required to visualise potential residual tumour cells and thereby improve treatment outcome for HNSCC and ESCC patients. We developed a device to spatiotemporally measure oxygen consumption rates (OCR) and extracellular acidification rates (ECAR) to distinguish HNSCC and ESCC cells from healthy cells in vitro, exploiting general metabolic differences between cancer cells and healthy cells., Methods: OCR and ECAR were measured via a newly developed device named STO 2 p-Q (SpatioTemporal O 2 and pH Quantification) using the VisiSens technology based on ratiometric fluorescence imaging, facilitating spatiotemporal resolution. Results were confirmed using extracellular flux analyses (Seahorse technology)., Results: STO 2 p-Q is described and used to measure OCR and ECAR in HNSCC and ESCC cell lines and normal fibroblast and epithelial cells as components of the tumour microenvironment. OCR measurements showed differences amongst HNSCC and ESCC cell lines and between HNSCC/ESCC and normal cells, which on average had lower OCR than HNSCC/ESCC cells. Both OCR and ECAR measurements were independently verified using the Seahorse technology. Additionally, using STO 2 p-Q, HNSCC/ESCC, and normal cells could be spatially resolved with a resolution in the low millimetre range., Conclusions: We developed a method to spatiotemporally measure OCR and ECAR of cells, which has many potential in vitro applications and lays the foundation for the development of novel diagnostic tools for the detection of cancerous tissue in HNSCC and ESCC patients in vivo.

Microphysiometry is a powerful technique to study metabolic parameters and detect changes to external stimuli. However, applying this technique for automated label-free and real-time measurements within cell-laden three-dimensional (3D) cell culture constructs remains a challenge. Herein, we present an entirely automated microphysiometry setup that combines needle-type microsensors with motorized sample and sensor positioning systems inside a standard tissue-culture incubator. The setup records dissolved oxygen as a metabolic parameter along the z -direction within cell-laden 3D constructs in a minimally invasive manner. The microphysiometry setup was applied to characterize the spatial oxygen distribution within thick cell-laden 3D constructs, study the time-dependent changes on the oxygen tension within 3D breast cancer models following a chemotherapeutic treatment, and identify kinetics and recovery effects after drug exposure over 5 weeks. Our data suggest that the microphysiometry setup enables highly reproducible measurements without human intervention, due to the high degree of automation and positional accuracy. The results demonstrate the applicability of the setup to provide valuable long-term insights into oxygenation within 3D models using minimally invasive, label-free, and entirely automated analysis methods.

Oxygen tension plays an important role in overall cell function and fate, regulating gene expression, and cell differentiation. Although there is extensive literature available that supports the previous statement, little information is to be found about accurate O 2 measurements during culture. In fact, O 2 concentration at the cell layer during culture is commonly assumed to be equal to that of the incubator atmosphere. This assumption does not consider oxygen diffusion properties, cell type, cell density, media composition, time in culture nor height of the cell culture medium column. In this study, we developed a non-invasive, optical sensor foil-based technique suitable for measuring the 3D oxygen gradient that is formed during cell culture as a result of normal cell respiration. For this propose, we created a 3D printed ramp to which surface an oxygen optode sensor foil was attached. The ramps were positioned inside the culture wells of 24 well plate prior cell seeding. This set up in conjunction with the VisiSens TD camera system allows to investigate the oxygen gradient formation during culture. Cultivation was performed with three different initial cell densities of the cell line A549 that were seeded on the plate containing the ramps with the oxygen sensors. The O 2 gradient obtained after 96 h of culture showed significantly lower O 2 concentrations closer to the bottom of the well in high cell density cultures compared to that of lower cell density cultures. Furthermore, it was very interesting to observe that even with low cell density culture, oxygen concentration near the cell layer was lower than that of the incubator atmosphere. The obtained oxygen gradient after 96 h was used to calculate the oxygen consumption rate (OCR) of the A549 cells, and the obtained value of ~100 fmol/h/cell matches the OCR value already reported in the literature for this cell line. Moreover, we found our set up to be unique in its ability to measure oxygen gradient formation in several wells of a cell culture plate simultaneously and in a non-invasive manner., (Copyright © 2020 Peniche Silva, Liebsch, Meier, Gutbrod, Balmayor and van Griensven.)

Tissue engineering (TE) approaches typically envisage the structural and functional reconstitution of previously damaged tissue in situ. An adequate three-dimensional environment is therefore of fundamental importance for the designated cells associated to the scaffold material. The sufficient supply with nutrients and oxygen in vitro and in vivo mark thereby critical challenges of TE. In this study, we intended to analyse the level of locally dissolved oxygen within 3-D cell-loaded collagen I gels in vitro. For the analysis of the oxygen levels in situ, we employed an optical fibre-based micro sensor setup, as well as a camera supported non-invasive optical sensor foil based technique. These complementary analytical tools enable the identification, localization, and temporal follow-up investigation of specified regions of interest within TE constructs. Human adipose-derived mesenchymal stem cells (hAdMSCs) cultured in collagen I gels under normoxic conditions were analysed periodically and kinetically up to 70 days - thereby revealing dynamic changes of the level of dissolved oxygen inside the gel constructs. Dependent on the applied cell concentration, the in vitro oxygen concentration (cO 2 ) within the gels reached physiological ranges (7-9%) after 21 days, or 35 days of culture. The minimal cO 2 was measured after 35 days in vitro, featuring an oxygen level of 4.8 ± 1.3%. Upon prolonged culture, a plateau-like status of the cO 2 around 8-9% established, indicating a change in the physiological activity of the cells under investigation. The expression patterns of BCL2, CASP3 and MCM5 revealed significant differences among the proliferative and apoptotic stages of the cell-loaded samples at the investigated time points of 7 and 70 days in culture. In summary, these data show the temporary dynamic nature of the oxygen distribution in cell-loaded gel constructs. The applied technique is an ideal tool for the evaluation of multiple parameters affecting the oxygen distribution in vitro. We conclude that it takes 5 weeks for establishing an equilibrium of cO 2 . Levels reached in a 3-D gel construct are comparable with physiological oxygenation ranges in bone-associated tissues., (Copyright © 2018. Published by Elsevier B.V.)

In many wetland plants, belowground transport of O 2 via aerenchyma tissue and subsequent O 2 loss across root surfaces generates small oxic root zones at depth in the rhizosphere with important consequences for carbon and nutrient cycling. This study demonstrates how roots of the intertidal salt-marsh plant Spartina anglica affect not only O 2 , but also pH and CO 2 dynamics, resulting in distinct gradients of O 2 , pH, and CO 2 in the rhizosphere. A novel planar optode system (VisiSens TD ® , PreSens GmbH) was used for taking high-resolution 2D-images of the O 2 , pH, and CO 2 distribution around roots during alternating light-dark cycles. Belowground sediment oxygenation was detected in the immediate vicinity of the roots, resulting in oxic root zones with a 1.7 mm radius from the root surface. CO 2 accumulated around the roots, reaching a concentration up to threefold higher than the background concentration, and generally affected a larger area within a radius of 12.6 mm from the root surface. This contributed to a lowering of pH by 0.6 units around the roots. The O 2 , pH, and CO 2 distribution was recorded on the same individual roots over diurnal light cycles in order to investigate the interlinkage between sediment oxygenation and CO 2 and pH patterns. In the rhizosphere, oxic root zones showed higher oxygen concentrations during illumination of the aboveground biomass. In darkness, intraspecific differences were observed, where some plants maintained oxic root zones in darkness, while others did not. However, the temporal variation in sediment oxygenation was not reflected in the temporal variations of pH and CO 2 around the roots, which were unaffected by changing light conditions at all times. This demonstrates that plant-mediated sediment oxygenation fueling microbial decomposition and chemical oxidation has limited impact on the dynamics of pH and CO 2 in S. anglica rhizospheres, which may in turn be controlled by other processes such as root respiration and root exudation.

Root-mediated CO 2 uptake, O 2 release and their effects on O 2 and CO 2 dynamics in the rhizosphere of Lobelia dortmanna were investigated. Novel planar optode technology, imaging CO 2 and O 2 distribution around single roots, provided insights into the spatiotemporal patterns of gas exchange between roots, sediment and microbial community. In light, O 2 release and CO 2 uptake were pronounced, resulting in a distinct oxygenated zone (radius: c. 3 mm) and a CO 2 -depleted zone (radius: c. 2 mm) around roots. Simultaneously, however, microbial CO 2 production was stimulated within a larger zone around the roots (radius: c. 10 mm). This gave rise to a distinct pattern with a CO 2 minimum at the root surface and a CO 2 maximum c. 2 mm away from the root. In darkness, CO 2 uptake ceased, and the CO 2 -depleted zone disappeared within 2 h. By contrast, the oxygenated root zone remained even after 8 h, but diminished markedly over time. A tight coupling between photosynthetic processes and the spatiotemporal dynamics of O 2 and CO 2 in the rhizosphere of Lobelia was demonstrated, and we suggest that O 2 -induced stimulation of the microbial community in the sediment increases the supply of inorganic carbon for photosynthesis by building up a CO 2 reservoir in the rhizosphere., (© 2018 The Authors. New Phytologist © 2018 New Phytologist Trust.)

Bone tissue engineering (BTE) utilizing biomaterial scaffolds and human mesenchymal stem cells (hMSCs) is a promising approach for the treatment of bone defects. The quality of engineered tissue is crucially affected by numerous parameters including cell density and the oxygen supply. In this study, a novel oxygen-imaging sensor was introduced to monitor the oxygen distribution in three dimensional (3D) scaffolds in order to analyze a new cell-seeding strategy. Immortalized hMSCs, pre-cultured in a monolayer for 30-40% or 70-80% confluence, were used to seed demineralized bone matrix (DBM) scaffolds. Real-time measurements of oxygen consumption in vitro were simultaneously performed by the novel planar sensor and a conventional needle-type sensor over 24 h. Recorded oxygen maps of the novel planar sensor revealed that scaffolds, seeded with hMSCs harvested at lower densities (30-40% confluence), exhibited rapid exponential oxygen consumption profile. In contrast, harvesting cells at higher densities (70-80% confluence) resulted in a very slow, almost linear, oxygen decrease due to gradual achieving the stationary growth phase. In conclusion, it could be shown that not only the seeding density on a scaffold, but also the cell density at the time point of harvest is of major importance for BTE. The new cell seeding strategy of harvested MSCs at low density during its log phase could be a useful strategy for an early in vivo implantation of cell-seeded scaffolds after a shorter in vitro culture period. Furthermore, the novel oxygen imaging sensor enables a continuous, two-dimensional, quick and convenient to handle oxygen mapping for the development and optimization of tissue engineered scaffolds. Biotechnol. Bioeng. 2017;114: 894-902. © 2016 Wiley Periodicals, Inc., (© 2016 Wiley Periodicals, Inc.)

The high-throughput analysis of respiratory activity has become an important component of many biological investigations. Here, a technological platform, denoted the "MultiSense tool," is described. The tool enables the parallel monitoring of respiration in 100 samples over an extended time period, by dynamically tracking the concentrations of oxygen (O 2 ) and/or carbon dioxide (CO 2 ) and/or pH within an airtight vial. Its flexible design supports the quantification of respiration based on either oxygen consumption or carbon dioxide release, thereby allowing for the determination of the physiologically significant respiratory quotient (the ratio between the quantities of CO 2 released and the O 2 consumed). It requires an LED light source to be mounted above the sample, together with a CCD camera system, adjusted to enable the capture of analyte-specific wavelengths, and fluorescent sensor spots inserted into the sample vial. Here, a demonstration is given of the use of the MultiSense tool to quantify respiration in imbibing plant seeds, for which an appropriate step-by-step protocol is provided. The technology can be easily adapted for a wide range of applications, including the monitoring of gas exchange in any kind of liquid culture system (algae, embryo and tissue culture, cell suspensions, microbial cultures).

Conventional oxygen (micro-) sensors assess oxygen concentration within a particular region or across a transect of tissue, but provide no information regarding its bidimensional distribution. Here, a novel imaging technology is presented, in which an optical sensor foil (i.e., the planar optode) is attached to the surface of the sample. The sensor converts a fluorescent signal into an oxygen value. Since each single image captures an entire area of the sample surface, the system is able to deduce the distribution of oxygen at a resolution level of few micrometers. It can be deployed to dynamically monitor oxygen consumption, thereby providing a detailed respiration map at close to cellular resolution. Here, we demonstrate the application of the imaging tool to developing plant seeds; the protocol is explained step by step and some potential pitfalls are discussed.

In Salmonella infection, the Salmonella pathogenicity island-2 (SPI-2)-encoded type three secretion system (T3SS2) is of key importance for systemic disease and survival in host cells. For instance, in the streptomycin-pretreated mouse model SPI-2-dependent Salmonella replication in lamina propria CD11c(-)CXCR1(-) monocytic phagocytes/macrophages (MΦ) is required for the development of colitis. In addition, containment of intracellular Salmonella in the gut critically depends on the antimicrobial effects of the phagocyte NADPH oxidase (PHOX), and possibly type 2 nitric oxide synthase (NOS2). For both antimicrobial enzyme complexes, oxygen is an essential substrate. However, the amount of available oxygen upon enteroinvasive Salmonella infection in the gut tissue and its impact on Salmonella-MΦ interactions was unknown. Therefore, we measured the gut tissue oxygen levels in a model of Salmonella enterocolitis using luminescence two-dimensional in vivo oxygen imaging. We found that gut tissue oxygen levels dropped from ∼78 Torr (∼11% O2) to values of ∼16 Torr (∼2% O2) during infection. Because in vivo virulence of Salmonella depends on the Salmonella survival in MΦ, Salmonella-MΦ interaction was analysed under such low oxygen values. These experiments revealed an increased intracellular replication and survival of wild-type and t3ss2 non-expressing Salmonella. These findings were paralleled by blunted nitric oxide and reactive oxygen species (ROS) production and reduced Salmonella ROS perception. In addition, hypoxia enhanced SPI-2 transcription and translocation of SPI-2-encoded virulence protein. Neither pharmacological blockade of PHOX and NOS2 nor impairment of T3SS2 virulence function alone mimicked the effect of hypoxia on Salmonella replication under normoxic conditions. However, if t3ss2 non-expressing Salmonella were used, hypoxia did not further enhance Salmonella recovery in a PHOX and NOS2-deficient situation. Hence, these data suggest that hypoxia-induced impairment of antimicrobial activity and Salmonella virulence cooperate to allow for enhanced Salmonella replication in MΦ., (© 2015 John Wiley & Sons Ltd.)

Tissue oxygenation plays a critical role in the pathogenesis of various diseases, but non-invasive, robust and user-friendly methods for its measurement in vivo still need to be established. Here, we are presenting an in vivo oxygen-detection system that uses ratiometric luminescence imaging (RLI) as a readout scheme to determine the skin oxygen tension of mouse hind footpads via side-by-side comparison with more established techniques including luminescence-lifetime imaging using planar sensor films and the polarographic electrode as the gold standard. We also demonstrate that this technology allows the detection of changes in mouse skin tissue oxygenation induced by subjecting mice to systemic hypoxia. The data demonstrate oxygen imaging based on RLI to be a most useful tool for reliably and easily analyzing and monitoring skin tissue oxygenation in vivo. This technology will advance our understanding of local regulation of skin tissue oxygenation in various disease conditions.

Background and Aims: Live imaging methods have become extremely important for the exploration of biological processes. In particular, non-invasive measurement techniques are key to unravelling organism-environment interactions in close-to-natural set-ups, e.g. in the highly heterogeneous and difficult-to-probe environment of plant roots: the rhizosphere. pH and CO2 concentration are the main drivers of rhizosphere processes. Being able to monitor these parameters at high spatio-temporal resolution is of utmost importance for relevant interpretation of the underlying processes, especially in the complex environment of non-sterile plant-soil systems. This study introduces the application of easy-to-use planar optode systems in different set-ups to quantify plant root-soil interactions., Methods: pH- and recently developed CO2-sensors were applied to rhizobox systems to investigate roots with different functional traits, highlighting the potential of these tools. Continuous and highly resolved real-time measurements were made of the pH dynamics around Triticum turgidum durum (durum wheat) roots, Cicer arietinum (chickpea) roots and nodules, and CO2 dynamics in the rhizosphere of Viminaria juncea., Key Results: Wheat root tips acidified slightly, while their root hair zone alkalized their rhizosphere by more than 1 pH unit and the effect of irrigation on soil pH could be visualized as well. Chickpea roots and nodules acidified the surrounding soil during N2 fixation and showed diurnal changes in acidification activity. A growing root of V. juncea exhibited a large zone of influence (mm) on soil CO2 content and therefore on its biogeochemical surrounding, all contributing to the extreme complexity of the root-soil interactions., Conclusions: This technique provides a unique tool for future root research applications and overcomes limitations of previous systems by creating quantitative maps without, for example, interpolation and time delays between single data points.

Background: In a preliminary trial, we were able to show first promising results in the analysis of perioperative and postoperative perfusion of free flaps by means of a new monitoring system for detecting thrombotic vessel occlusion before clinical signs become evident., Objective: We investigated whether flap monitoring by measuring perfusion-dependent parameters differs between radial forearm and fibular free flaps and whether a threshold value requiring anastomosis revision could be determined., Methods: 37 radial forearm flaps (RF) and 15 fibular flaps (FF) were harvested and transplanted. Perfusion was determined by measuring a fluorescent oxygen sensor foil covering a flap's skin surface with a handheld fluorescence microscope. The sensor contained an oxygen reservoir, which was consumed by the tissue corresponding to the perfusion status of the flap. Measurements were done before explantation, after successful anastomosis and one day after surgery., Results: We found a significant difference (p < 0.005) in the relative transdermal oxygen consumption (RTOC) between clinically well-perfused grafts (RF: mean: 0.13 ± 0.08; FF: mean: 0.15 ± 0.07) and clinically poorly perfused grafts (RF: mean: 0.40 ± 0.09; FF: mean: 0.55 ± 0.28). A threshold RTOC value of 0.3 for differentiating between well-perfused and poorly perfused flaps was confirmed for both RF and FF.

Biological samples are far from homogeneous, with complex compartmentation being the norm. Major physiological processes such as respiration do not therefore occur in a uniform manner within most tissues, and it is currently not possible to image its gradients in living plant tissues. A compact fluorescence ratiometric-based device is presented here, consisting of an oxygen-sensitive foil and a USB (universal serial bus) microscope. The sensor foil is placed on the sample surface and, based on the localized change in fluorescence signal over time, information about the oxygen consumption (respiration) or evolution (photosynthesis) can be obtained. Using this imaging technique, it was possible to demonstrate the spatial pattern of oxygen production and consumption at a c. 20-μm level of resolution, and their visualization in the rhizosphere, stem and leaf, and within the developing seed. The oxygen mapping highlighted the vascular tissues as the major stem sink for oxygen. In the leaf, the level of spatial resolution was sufficient to visualize the gas exchange in individual stomata. We conclude that the novel sensor set-up can visualize gradients in oxygen-consuming and producing processes, thereby facilitating the study of the spatial dynamics of respiration and photosynthesis in heterogeneous plant tissues., (© 2012 The Authors. New Phytologist © 2012 New Phytologist Trust.)

Monitoring p(bt)O(2) is a valuable supplemental -procedure for neurocritically ill patients. Here, we utilise an opto-chemical method for measuring cortical pO(2) during a reversibly introduced epidural mass lesion in a rat model. The sensor was placed in a cortical window of 17 ventilated Wistar rats. Inflating the balloon device over the contralateral hemisphere increased ICP. Physiological parameters and cortical pO(2) were recorded. The ICP increased from 6.2 ± 4.6 to 44.6 ± 12.6 mmHg (p < 0.001). Cortical pO(2) over arterioles changed from 28.9 ± 2.1 to 19.0 ± 2.1 mmHg (p < 0.001), over venules from 14.8 ± 1.2 to 9.9 ± 1.5 mmHg (p = 0.002) and over parenchyma from 4.1 ± 0.7 to 2.4 ± 0.8 mmHg respectively (p < 0.001), while basic physiological parameters remained constant (p > 0.05). During baseline, arterial pO(2) correlated significantly with cortex arteriole and venole pO(2), but not with cortex parenchyma pO(2). While ICP was raised, cortical pO(2) did not correlate with arterial pO(2). In this model, a moderate epidural mass lesion causes a significant decrease in cortical pO(2). Cortex parenchyma pO(2) appeared to be independent from arterial pO(2). The correlation of cortex vessel pO(2) with arterial pO(2) disappeared during the epidural mass lesion. These findings show the capability of the device to elucidate the behaviour of functionally different cortex areas at pathophysiological conditions.

Background: Free tissue transfer in head and neck reconstructions has a very high success rate, but thrombotic vessel occlusion is still a serious complication occurring in up to 10% of all cases. Thus, a simple, fast and reliable monitoring system for free flaps would be of advantage., Objective: The aim of this study was to investigate whether free flap monitoring by measuring perfusion-dependent parameters is a suitable method for discovering vessel thrombosis in free flaps., Methods: 10 patients requiring tissue reconstruction after tumour surgery or because of chronic wounds were included in this study. 10 microvascular flaps were harvested and transplanted. Perfusion was determined by measuring a fluorescent oxygen sensor foil covering the flap's skin surface by means of a USB-handheld fluorescence microscope prototype. The sensor contained an oxygen reservoir which was consumed by the tissue corresponding to the perfusion status of the flap. Measurements were done before explantation, after successful anastomosis and 1 day after surgery., Results: Clinically well-perfused grafts showed slope values between 0.07 and 0.27 (mean: 0.18 ± 0.07), and clinically poorly perfused grafts showed slope values between 0.35 and 0.75 (mean: 0.52 ± 0.19). In the present study, we used a threshold slope value of 0.3 for differentiating between well-perfused and poorly perfused flaps., Conclusion: Flap monitoring via oxygen imaging by means of fluorescent sensor foils appears to be a fast, non-invasive, cost-effective and thus suitable method for analyzing flap perfusion with the additional advantage of aiding decision making on flap revision.

Purpose: We present the results of the visualisation of radial oxygen gradients in rats' cortices and their potential use in neurocritical management., Methods: PO₂ maps of the cortex of ten sedated, intubated and controlled ventilated Wistar rats were obtained with a camera (SensiMOD, PCO, Kelheim, Germany). Those pictures were analysed and edited by a custom-made software. A virtual matrix, designed to evaluate the cortical O₂ partial pressure, was placed vertically to the artery under investigation, and afterwards multiple regions of interest were measured (width 10 pixels, length 15-50 pixels). The results showed a map of the cerebral oxygenation, which allowed us to calculate radial oxygen gradients over arterioles. Three groups were defined according to the level of the arterial pO₂: PaO₂ < 80, PaO₂ 80-120 and PaO₂ > 120. Gradients were analysed from the middle of the vessel to its border (1), from the border into the parenchyma next to the vessel (2) and a combination of both (3)., Results: Gradient 1 showed significantly different cortical pO₂ values between the three different groups. The mean pO₂ values were 2.62, 5.29 and 5.82 mmHg/mm. Gradient 2 measured 0.56, 0.90 and 1.02 mmHg/mm respectively. Gradient 3 showed significant results between the groups with values of 3.18, 6.19 and 6.84 mmHg/mm., Conclusion: Using these gradients, it is possible to describe and compare the distribution of oxygen to the brain parenchyma. With the presented technique, it is possible to detect pO₂ changes in the oxygen supply of the brain cortex.

We present a non invasive fluorescein based method to measure and visualise the partial oxygen pressure of the rat cortex in a 2D picture. We studied 10 Wistar rats. A trepanation was done over the hemisphere and the dura was opened. A PMMA cylinder with a calibrated optical membrane was fixed over the surface of the brain. The CCD camera with the light source is placed over the cylinder. This allows the generation of two-dimensional maps of the pO2 pressure. Using the white light picture we defined regions of interest (ROI) in an artery, vein, parenchyma and an overall ROI. For every ROI a mean emission value was calculated. We increased, stepwise, the FiO2 from 30% up to 100%. Thereafter we established ventilation with an FiO2 of 30% and induced a stepwise hypo- and hyperventilation. The ROI's showed significantly different pO2 values. The apO2 showed a good correlation to the pO2 in the ROIs. This new set up seems to give reliable absolute pO2 values of the brain surface. This method seems to be able for the first time to give a non invasive pO2 map of the brain surface reflecting oxygenation and ventilation effects.

Background: Oxygen-dependent quenching of luminescence using transparent planar sensor foils was shown to overcome the limitations of the polarographic electrode technique in an animal model. This method was then transferred to a clinical setting to measure the transcutaneous pO(2) (p(tc)O(2))., Methods: In six healthy subjects, a cuff on the upper arm was occluded up to 20 mmHg above systolic pressure and released after 8 min. P(tc)O(2) was measured at the lower arm every 30 s before, during, and up to 20 min after cuff occlusion (40 degrees C applied skin temperature) using luminescence lifetime imaging (LLI) of platinum(II)-octaethyl-porphyrin immobilized in a polystyrene matrix. For validation, the polarographic Clark electrode technique was applied in close proximity, and measurements were conducted simultaneously., Results: P(tc)O(2) measurements before (70.8+/-19.1 vs. 66.2+/-7.7 mmHg) and at the end of ischemic (2.7+/-1.2 vs. 3.6+/-1.7 mmHg) and reperfusion phases (72.2+/-3.6 vs. 68.4+/-8.9 mmHg) did not differ significantly using the Clark electrode vs. LLI. At both the initial ischemic and the reperfusion phases, the Clark electrode measured a faster decrease or increase, respectively, in p(tc)O(2) because of the oxygen consumption occurring in this method., Conclusion: The presented method provides accurate and reproducible p(tc)O(2) values under changing microcirculatory conditions. The lack of oxygen consumption during measurement allows both a more realistic estimation of p(tc)O(2) than compared with the gold standard and permanent use in regions with critical oxygen supply.

Background/purpose: Reliable information of cerebral oxygenation is-besides the monitoring of the intracranial pressure-of eminent interest when treating patients with brain injuries. In this study, we introduce a new, fast, and sensitive method capable of determining the cortical partial oxygen pressure on the surface of the cortex using a special sensor foil., Methods: The introduced method exploits the O2-dependent phosphorescence of a thin sensor foil, which is excited by a short light-emitting diode flash. The optical signal is registered by a charge-coupled device camera and analyzed with PC-based software. The adequacy of this method was tested in 10 animals. The sensor device was placed directly over the cortex after craniotomy and removal of the dura. Arterial oxygen pressure was systematically varied by modifying the ventilation gas mixture. A total of 225 measurements were performed within 4 regions of interest., Results: Obtained results were sufficient in each case. The pO2 over the cortex correlated well with arterial pO2. Measurements over arteries showed a correlation coefficient of 0.72 (P<0.001), over veins 0.58 (P<0.001), over cortical parenchyma 0.46 (P<0.001), and in a larger region of interest containing vessels and cortical tissue 0.59 (P<0.001). The frequency of the measurements was 7 Hz with a single measurement covering an area of 30 x 30 microm., Conclusions: For the first time, nearly online pO2 maps of a brain cortex can be generated, allowing simultaneously also separate measurements over distinct anatomic structures yielding a good spatial resolution.

Background: Measuring brain oxygenation in patients with TBI or SAH is of major interest. We present a new semi-invasive method for two dimensional measurements of cortical pO2., Methods: For this feasibility study, a porphyrin containing sensor foil was placed directly on the cortex of intubated and variably ventilated Wistar rats. The sensor was excited with a light pulse and pictures of the foil's pO2 dependant emissions were captured with a CCD camera. After online data processing, two-dimensional maps of cortex oxygenation were displayed and analyzed using ROIs (here: arteriole, vein, parenchyma) with a display rate of 7 Hz. The size of one single measurement pixel was 0.03 x 0.03 mm2., Findings: The mean pO2 over cortex arterioles was 20.3 +/- 0.69, over veins 17.1 +/- 0.5 and over parenchyma 9.1 +/- 0.6 (mmHg +/- SD). The arterial pO2 showed a good correlation to the pO2 in the ROIs (r = 0.46-0.72, p < 0.0001, n = 198). Comparing groups with different paO2 and paCO2 we found significantly different pO2 values in the ROIs of the cortex., Conclusions: This prototype is capable of obtaining cortical pO2 maps with excellent temporal and spatial resolution and provides simultaneous imaging of the cortex structures.

Objective: Oxygen-dependent quenching of luminescence of metal porphyrin complexes has been used to image the pO(2) distribution over tumor and normal tissue., Methods: An experimental setup is described using a platinum(II)-octaethyl-porphyrin immobilized in a polystyrene matrix as transparent planar sensor., Results: Sensitivity over a broad range is high at low pO(2) values (+/- 0.2 mm Hg at 0 mm Hg; +/- 1.5 mm Hg at 160 mm Hg pO(2)). Due to intrinsically referencing via lifetime encoding there was no modification of the sensor response in vivo in the dorsal skinfold chamber model with amelanotic melanoma (A-MEL-3) in awake hamsters when compared to the in vitro calibration. pO(2) measurements over normal tissue (25.8 +/- 5.1 mm Hg) and tumor tissue (9.2 +/- 5.1 mm Hg) were in excellent agreement with previous results obtained in this model using a surface multiwire electrode., Conclusions: Using the presented method the surface pO(2) distribution can be mapped with a high temporal resolution of approximately 100 ms and a spatial resolution of at least 25 mu m. Moreover, the transparent sensor allows the simultaneous visualization of the underlying microvasculature.

A cross-reactive array in a micro titer plate (MTP) format is described that is based on a versatile and highly flexible scheme. It makes use of rather unspecific metal ions probes having almost identical fluorescence spectra, thus enabling (a) interrogation at identical analytical wavelengths, and (b) imaging of the probes contained in the wells of the MTP using a CCD camera and an array of blue-light-emitting diodes as a light source. The unselective response of the indicators in the presence of mixtures of five divalent cations generates a characteristic pattern that was analyzed by chemometric tools. The fluorescence intensity of the indicators was transferred into a time-dependent parameter applying a scheme called dual lifetime referencing. In this method, the fluorescence decay profile of the indicator is referenced against the phosphorescence of an inert reference dye added to the system. The intrinsically referenced measurements also were performed using blue LEDs as light sources and a CCD camera without intensifiers as the detector. The best performance was observed if each well was excited by a single LED. The assembly allows the detection of dye concentrations in the nanomoles-per-liter range without amplification and the acquisition of 96 wells simultaneously. The pictures obtained form the basis for evaluation by pattern recognition algorithms. Support vector machines are capable of predicting the presence of significant concentrations of metal ions with high accuracy.

To improve efficacy of photodynamic therapy (PDT) with intravenously administered 5-aminolaevulinic acid (ALA) fractionating the light dose or reducing the light intensity may be a possibility. Therefore, Syrian Golden hamsters were fitted with dorsal skinfold chambers containing an amelanotic melanoma (n=26). PDT was performed (100 mW cm(-2), 100 J cm(-2), continuously or fractionated, and 25 mW cm(-2), 100 J cm(-2); continuously or fractionated) using an incoherent light source following i.v. application of ALA. Following fractionated irradiation, the light was paused after 20 J cm(-2) for 15 min. Prior to and up to 24 h after PDT tissue, pO(2) was measured using luminescence lifetime imaging. The efficacy was evaluated by measuring the tumour volume of amelanotic melanoma cells grown subcutaneously in the back of Syrian Golden hamsters (n=36). Only high-dose PDT resulted in a significant decrease of pO(2). Irrespective of the mode of irradiation only high-dose PDT induced complete remission of all tumours (13 out of 13). It could be shown that low-dose PDT failed to induce a significant decrease of pO(2). No significant effect of fractionated irradiation was shown regarding the therapeutic efficacy 28 days after PDT. Thus performing a fractionated PDT with ALA or reducing the light intensity seems not to be successful in clinical PDT according to the present data.

Nutrient and oxygen supply of cells are crucial to tissue engineering in general. If a sufficient supply cannot be maintained, the development of the tissue will slow down or even fail completely. Previous studies on oxygen supply have focused on measurement of oxygen partial pressures (pO(2)) in culture media or described the use of invasive techniques with spatially limited resolution. The experimental setup described here allows for continuous, noninvasive, high-resolution pO(2) measurements over the cross-section of cultivated tissues. Applying a recently developed technique for time-resolved pO(2) sensing using optical sensor foils, containing luminescent O(2)-sensitive indicator dyes, we were able to monitor and analyze gradients in the oxygen supply in a tissue over a 3-week culture period. Cylindrical tissue samples were immobilized on top of the sensors. By measuring the luminescence decay time, two-dimensional pO(2) distributions across the tissue section in contact with the foil surface were determined. We applied this technique to cartilage explants and to tissue-engineered cartilage. For both tissue types, changes were detected in monotonously decreasing gradients of pO(2) from the surface with high pO(2) to minimum pO(2) values in the center of the samples. Nearly anoxic conditions were observed in tissue constructs ( approximately 0 Torr) but not in excised cartilage discs ( approximately 20 Torr) after 1 day. Furthermore, the oxygen supply seemed to strongly depend on cell density and cell function. Additionally, histological analysis revealed a maximum depth of approximately 1.3 mm of regular cartilage development in constructs grown under the applied culture conditions. Correlating analytical and histological analysis with the oxygen distributions, we found that pO(2) values below 11 Torr might impair proper tissue development in the center. The results illustrate that the method developed is an ideal one to precisely assess the oxygen demand of cartilage cultures., (Copyright 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 80: 73-83, 2002.)

We present a referenced scheme for fluorescence intensity measurements that is useful for imaging applications. It is based on the conversion of the fluorescence intensity information into a time-dependent parameter. A phosphorescent dye is added in the form of approximately 10-microm particles to the sample containing the pH-sensitive fluorescent indicator. Both the reference dye and the pH probe are excited simultaneously by a blue LED, and an overall luminescence is measured. In the time-resolved imaging method presented here, two images taken at different time gates were recorded using a CCD camera. The first image is recorded during excitation and reflects the luminescence signal of both the fluorophore (pH) and the phosphor (reference). The second image, which is measured after a certain delay (after switching off the light source), is solely caused by the long-lived phosphorescent dye. Because the intensity of the fluorophore contains the information on pH, whereas phosphorescence is pH-independent, the ratio of the images displays a referenced intensity distribution that reflects the pH at each picture element (pixel). The scheme is useful for LED light sources and CCD cameras that can be gated with square pulses in the microsecond range. The fundamentals and potential of this new method, to which we refer as time domain dual lifetime referencing (t-DLR), are demonstrated.

The purpose of this study was to investigate the functional role the two corticotropin-releasing hormone (CRH) receptor subtypes play in regulating the behavioural performance of rats in various well-defined test situations. Antisense oligodeoxynucleotides (ODNs) corresponding to either the rat CRH1 or CRH2 receptor mRNA were infused chronically into the lateral ventricle of male rats via osmotic minipumps (5 microg/0.5 microl/h over 6 days). Control groups received infusions of either a scrambled sequence ODN or mixed bases ODN or vehicle. On day 4 after surgery, the rats were subjected to 10 min of social defeat and immediately afterwards tested on the elevated plus-maze. Compared to a scrambled sequence control ODN, CRH1 receptor antisense ODN infusion was found to exert an anxiolytic-like effect whereas CRH2 receptor antisense ODN infusion had no effect on defeat-induced anxiety-related behaviour. In contrast, the CRH2 receptor antisense ODN increased immobility in a forced swim test whereas CRH1 receptor ODN-treated rats did not differ from controls. No influence of either ODN was found on general locomotor activity in an open field or on short-term memory performance in a social discrimination test. Furthermore, the CRH2 receptor antisense ODN did not affect spatial learning in a Morris water maze task. An additional experiment comparing a mixture of both missense ODNs and a vehicle control group confirmed that the former failed to induce non-specific (toxic) side effects, further substantiating the specificity of the respective antisense effects measured in this study. The results support the hypothesis that the two CRH receptor subtypes selectively mediate differential effects of endogenous CRH or CRH-related peptides at the brain level with the CRHI receptor contributing predominantly to emotional behaviour and the CRH2 receptor being involved in the regulation of stress coping behaviour.

Two Wistar rat lines, selectively bred for high-anxiety-related behavior (HAB) and low-anxiety-related behavior (LAB) in the elevated plus-maze test, were tested for the susceptibility of their behavioral characteristics to anxiolytic treatment and for their endocrine and physiological reactivity to different stressors. Injection of 1 mg/kg diazepam failed to affect line differences in coping strategy but resulted in a marked (20-fold) decrease in plus-maze anxiety in HAB rats; whereas, the anxiolytic effect was less pronounced in LAB animals. Biotelemetrical measurements revealed that HAB and LAB rats do not significantly differ in their baseline body temperature, locomotor activity, food and water intake, or in stress-induced alterations of the diurnal rhythms in these parameters. However, line differences were found in acute changes in body temperature and locomotor activity following stress exposure, LAB rats responding with a greater, albeit shorter, increase in body temperature and activity than HAB animals. Basal ACTH and corticosterone plasma levels as well as pituitary reactivity to intravenously administered CRH (40 ng/kg) were similar in both lines, although, especially in response to plus-maze exposure, HAB rats tended toward higher ACTH secretion than LAB rats. These data confirm that animals with high or low basal levels of anxiety may be a promising model for studying the mechanisms of action of anxiolytic substances. Nevertheless, the endocrine findings support the notion that the reactivity of the hypothalamo-pituitary-adrenocortical system and anxiety-related behavior can be regulated independently.

Over the past years, two breeding lines, derived originally from outbred Wistar rats, have been established that differ markedly and consistently in their anxiety-related behaviour in the elevated plus-maze. At the age of ten weeks, rats were tested once on the elevated plus-maze and the males and females displaying the most anxious and the least anxious behaviour were sib-mated to start a new generation of the high anxiety-related behaviour (HAB) and the low anxiety-related behaviour (LAB) lines, respectively. The resulting difference in emotionality between these two lines was also evident in an open field test and correlated with differences in the forced swim test. In the open field, the HAB rats tended to be less active and explored the central zone of the open field much less than the LAB animals. In the forced swim test, HAB rats started floating earlier, spent significantly more time in this immobile posture and struggled less than LAB rats. However, in an olfactory-cued social discrimination task there was no difference between male and female animals from either line. The overall performance in these various behavioural tests suggests that selective breeding has resulted in rat lines not only differing markedly in their innate anxiety-related behaviour in the plus-maze, but also in other stress-related behavioural performances, suggesting a close link between the emotional evaluation of a novel and stressful situation and an individual's coping strategy.

In order to test the hypothesis that prenatal hormones influence the emotional maturation of the offspring, the hypothalamo-pituitary-adrenal (HPA) axis activity was studied at the end of pregnancy in two rat breeding lines differing consistently in their innate anxiety-related behaviour in the elevated plus-maze. Virgin and pregnant rats were fitted with a chronic jugular vein catheter and tested 5 days later. The high basal level of anxiety-related behaviour (HAB) described in males and females of the HAB breeding line persists in pregnancy as indicated by a significantly reduced number of entries into and time spent on the open arms of the elevated plus-maze between days 18 and 20 of pregnancy compared with pregnant rats of the breeding line with low anxiety-related behaviour (LAB). In general, an increase in anxiety was found in both breeding lines in pregnancy compared with the respective virgin controls. With respect to HPA axis activity, increased basal levels of adrenocorticotropin (ACTH) and corticosterone have been found in pregnant rats of the HAB line compared with pregnant LAB rats. ACTH and corticosterone secretion in response to emotional and complex physical stressors (exposure to the elevated plus-maze and forced swimming, respectively) did not differ between virgin and pregnant rats of either breeding line. However, independent of the inborn emotionality of the animals, a general attenuation in the HPA axis response to stressors and to exogenous CRH could be confirmed in pregnant rats. The basal and stress-induced activity of the hypothalamo-neurohypophysial system secreting oxytocin and vasopressin was also tested, and no differences were found relating to the emotionality or reproductive state of the animals except for a reduced vasopressin secretion in pregnant HAB rats after forced swimming. The elevated basal activity of the HPA axis, including enhanced circulating concentrations of corticosterone in pregnant HAB rats, may influence both the neuroendocrine and emotional development of their offspring. Thus, the passing-on of maternal behavioural characteristics via prenatal, hormonal 'imprinting' has to be considered as a possible contribution to emotional maturation during an individual's development.

1. The responsiveness of the rat hypothalamo-pituitary-adrenal (HPA) axis and hypothalamo-neurohypophysial system (HNS) to emotional (elevated plus-maze) and physical (forced swimming) stressors and to administration of synthetic corticotrophin-releasing hormone (CRH) was investigated during pregnancy and lactation. In addition to pregnancy-related adaptations at the adenohypophysial level, behavioural responses accompanying the neuroendocrine changes were studied. 2. Whereas basal (a.m.) plasma corticosterone, but not corticotrophin (adrenocorticotrophic hormone; ACTH), levels were increased on the last day (i.e. on day 22) of pregnancy, the stress-induced rise in both plasma hormone concentrations was increasingly attenuated with the progression of pregnancy beginning on day 15 and reaching a minimum on day 21 compared with virgin control rats. A similar attenuation of responses to both emotional and physical stressors was found in lactating rats. 3. Although the basal plasma oxytocin concentration was elevated in late pregnancy, the stress-induced rise in oxytocin secretion was slightly lower in day 21 pregnant rats. In contrast to vasopressin, oxytocin secretion was increased by forced swimming in virgin and early pregnant rats indicating a differential stress response of these neurohypophysial hormones. 4. The blunted HPA response to stressful stimuli is partly due to alterations at the level of corticotrophs in the adenohypophysis, as ACTH secretion in response to CRH in vivo (40 ng kg-1, i.v.) was reduced with the progression of pregnancy and during lactation. In vitro measurement of cAMP levels in pituitary segments demonstrated reduced basal levels of cAMP and a lower increase after CRH stimulation (10 nM, 10 min) in day 21 pregnant compared with virgin rats, further indicating reduced corticotroph responsiveness to CRH in pregnancy. 5. The reduced pituitary response to CRH in late pregnancy is likely to be a consequence of a reduction in CRH receptor binding as revealed by receptor autoradiography. [125I] CRH binding in the anterior pituitary was significantly reduced in day 11, 17 and 22 pregnant rats compared with virgin controls. 6. Anxiety-related behaviour of the animals as revealed by the time on and entries into the open arms of the elevated plus-maze was different between virgin and pregnant rats with decreased number of entries indicating increased anxiety with the progression of pregnancy (except on pregnancy day 18). The emotional behaviour, however, was not correlated with the neuroendocrine responses. 7. The results indicate that the reduced response of the HPA axis to stressors described previously during lactation is already manifested around day 15 of pregnancy in the rat and involves physiological adaptations at the adenohypophysial level. However, alterations in stressor perception at higher brain levels with the progression of pregnancy may also be involved.

In order to study the dynamics of ethanol drinking behaviour, male Wistar rats were given the free choice between tap water, and 5, 10 and 20% ethanol solutions. After 8 weeks of continuous access, the animals were repeatedly deprived of the ethanol solutions for 3 days every 4 weeks. In the first experiment, drinking patterns were recorded for 24 h with an electronic drinkometer device, at different time-points of ethanol experience and after an ethanol deprivation episode. The preference for more highly concentrated ethanol solutions as well as ethanol consumption increased with continuing ethanol experience. Furthermore, after the ethanol deprivation episode, the animals immediately and preferentially drank from the 20% ethanol solution, the most highly concentrated ethanol solution offered. Additionally, the number of drinking bouts, particularly at the 10 and 20% ethanol solutions, was increased during the first hour after ethanol re-presentation. In a second experiment, the effects of repeated ethanol deprivation experience, inherent in this self-administration paradigm, on anxiety-related behaviour were tested on the elevated plus-maze. Repeated ethanol deprivation proved to be more anxiogenic than the first deprivation experience. Taken together, these findings suggest that ethanol deprivation is anxiogenic in long-term voluntarily ethanol-drinking rats, which is increased by repeated ethanol deprivation experience. The possibility that anxiety during ethanol deprivation might contribute to the 'relapse'-like drinking behaviour is discussed.

The effects of the central and peripheral administration of atriopeptin II, a 23-amino acid residue peptide of atrial natriuretic peptide (Ser103-Arg125) on anxiety-related behavior and on locomotor activity, was studied in male Wistar rats. Their behavior on the elevated plus-maze after social defeat stress indicated that intracerebroventricular (2.5 and 5 micrograms) and intraperitoneal (50 micrograms) administration of atriopeptin II produced anxiolysis. A low dose of 0.25 micrograms atriopeptin II administered bilaterally into the central nucleus of the amygdala was also found to be anxiolytic. Because intracerebroventricular administration of 5 micrograms atriopeptin II did not affect locomotor activity in the open-field test, the possibility that the anxiolytic effect was secondary to sedation could be ruled out. The anxiolytic effects observed after central and peripheral administration support the idea that atrial natriuretic peptide, which is increased in panic-anxiety, may be involved in the tapering of anxiety-related behavior.