Lipid redistribution due to a cell-cell fusion pore.

We consider the redistribution of lipids comprising the plasma membranes during cell-cell fusion, particularly due to the presence of a fusion pore. Assuming the membranes are of constant thickness, we find that the mole fraction of cholesterol increases in the directly apposed exoplasmic leaflets, and is decreased in the cytoplasmic leaflets. The redistribution of the phospholipids is obtained as well.

Lipid redistribution in the highly curved footprint of Piezo1.

We investigate the effects on the distribution of lipids in the plasma membrane that are caused by the insertion of a protein, Piezo1, that significantly distorts the membrane toward the cytosol. From coarse-grained molecular dynamics simulations, we find that the major effects occur in the outer, extracellular, leaflet. The mol fraction of cholesterol increases significantly in the curved region of the membrane close to Piezo1, while those of phosphatidylcholine and of sphingomyelin decrease. In the inner leaflet, mol fractions of cholesterol and of phosphatidylethanolamine decrease slightly as the protein is approached, while that of phosphatidylserine increases slightly. The mol fraction of phosphatidylcholine decreases markedly as the protein is approached. Most of these results are understood in the context of a theoretical model that utilizes two elements: 1) a coupling between the leaflets' actual curvatures and their compositionally dependent spontaneous curvatures and 2) the dependence of the spontaneous curvatures not only on the mol fractions of the phospholipids, but also on the effect that cholesterol has on the spontaneous curvatures of the phospholipids.

On the force between "rafts".

"Rafts" in the plasma membrane are nanoscopic domains rich in sphingolipids and cholesterol. Groups of them are thought to provide stable platforms for various proteins. How several rafts can form a larger stable platform has yet to be made clear. Here, we show that in the microemulsion theory of rafts, in which they, and the sea in which they float, are characterized by a naturally occurring length, rafts separated by a sea larger than this length will attract one another, thus making the sea smaller. Similarly, if rafts are separated by a sea that is smaller than the characteristic size, they will repel one another thus allowing the sea to increase in size. Such an elastic force can clearly stabilize a platform of many rafts.

A Theoretical Basis for Nanodomains.

We review the current theories of nanodomain, or "raft," formation. We emphasize that the idea that they are co-exisiting Lo and Ld phases is fraught with difficulties, as is the closely related idea that they are due to critical fluctuations. We then review an alternate theory that the plasma membrane is a two-dimensional microemulsion, and that the mechanism that drives to zero the line tension between Lo and Ld phases is the coupling of height and composition fluctuations. The theory yields rafts of SM and cholesterol in the outer leaf and POPS and POPC in the inner leaf. The "sea" between rafts consists of POPC in the outer leaf and POPE and cholesterol in the inner leaf. The characteristic size of the domain structures is tens of nanometers.

Model Plasma Membrane Exhibits a Microemulsion in Both Leaves Providing a Foundation for "Rafts".

We consider a model lipid plasma membrane, one that describes the outer leaf as consisting of sphingomyelin, phosphatidylcholine, and cholesterol and the inner leaf of phosphatidylethanolamine, phosphatidylserine, phosphatidylcholine, and cholesterol. Their relative compositions are taken from experiment; the cholesterol freely interchanges between leaves. Fluctuations in local composition are coupled to fluctuations in the local membrane curvature, as in the Leibler-Andelman mechanism. Structure factors of components in both leaves display a peak at nonzero wavevector. This indicates that the disordered fluid membrane is characterized by structure of the corresponding wavelength. The scale is given by membrane properties: its bending modulus and its surface tension, which arises from the membrane's connections to the cytoskeleton. From measurements on the plasma membrane, this scale is on the order of 100 nm. We find that the membrane can be divided into two different kinds of domains that differ not only in their composition but also in their curvature. The first domain in the outer, exoplasmic leaf is rich in cholesterol and sphingomyelin, whereas the inner, cytoplasmic leaf is rich in phosphatidylserine and phosphatidylcholine. The second kind of domain is rich in phosphatidylcholine in the outer leaf and in cholesterol and phosphatidylethanolamine in the inner leaf. The theory provides a tenable basis for the origin of structure in the plasma membrane and an illuminating picture of the organization of lipids therein.

Cholesterol-Dependent Bending Energy Is Important in Cholesterol Distribution of the Plasma Membrane.

We consider the plasma membrane that contains a cholesterol molar fraction of 0.4 and ask how that cholesterol is distributed between the two leaves. Because of the rapid flip-flop of cholesterol between leaves, we assume that its distribution is determined by the equality of its chemical potentials in the two leaves. When we consider only the contributions of entropy and interactions to the cholesterol chemical potential in our model system, we find, not surprisingly, that the cholesterol is mostly in the outer leaf because of the strong attraction between cholesterol and sphingomyelin (SM), which is predominantly in that leaf. We find 72% there. We then include the contribution from the bending energy in each leaf that must be overcome to join the leaves in a flat bilayer. The product of bending modulus and spontaneous curvature is obtained from simulation. We find that the addition of cholesterol to the outer leaf reduces the spontaneous curvature, which is initially positive, until it passes through zero when the molar fraction of cholesterol in the outer leaf is 0.28. Additional cholesterol is driven toward the inner leaf by the sphingomyelin phosphatidylcholine mixture. This is resisted by the bending energy contribution to the inner leaf. We find, again by simulation, that the addition of cholesterol monotonically increases the magnitude of the spontaneous curvature of the inner leaf, which is negative. This increases its bending energy. We conclude that, as a result of these competing effects, the percentage of cholesterol in the outer leaf is reduced to ∼63 ± 6%.

The Effect of Solutes on the Temperature of Miscibility Transitions in Multicomponent Membranes.

We address questions posed by experiments that show small-chain alcohols reduce the miscibility transition temperature when added to giant plasma membrane vesicles (GPMVs), but increase that temperature when added to giant unilamellar vesicles. In giant unilamellar vesicles the change in temperature displays a definite minimum, between decanol and tetradecanol, as a function of alcohol chain length; in GPMVs there is no such minimum. To emphasize the competition between internal entropies of the components and the interactions between them, we model the system as consisting of three different linear polymers. Two of them are the constituents of a liquid, one that can undergo a miscibility transition. To this liquid is added the third polymer component, which represents the short-chain alcohol. We show that, within Flory-Huggins theory, the addition of alcohol causes an increase or decrease of the miscibility transition temperature depending upon the competition of two effects. The first is the dilution of the interactions between the two components of the liquid caused by the introduction of the alcohol. This tends to lower the transition temperature. The second effect is the preferential partitioning of the alcohol into one phase of the liquid or the other. This tends to raise the transition temperature irrespective of which phase the alcohol prefers. This second effect is the smallest, and the decrease in transition temperature the largest, when the alcohol partitions equally between the two phases. Such equal partitioning occurs when the effect of the entropic excluded volume interactions (which cause the alcohol to prefer one phase) just balances the effect of the direct interactions, which cause it to prefer the other. These results allow us to make several predictions, and to propose an explanation for the different behavior of the transition temperature in GPMVs and giant unilamellar vesicles that results from the addition of alcohols.

[Phosphodiesterase 4 inhibition for treatment of endothelial barrier and microcirculation disorders in sepsis]. / Phosphodiesterase-4-Inhibition zur Therapie der endothelialen Schranken- und Mikrozirkulationsstörung in der Sepsis.

Sepsis is commonly associated with loss of microvascular endothelial barrier function (capillary leak) and dysfunctional microcirculation, which both promote organ failure. The development of a distinct therapy of impaired endothelial barrier function and disturbed microcirculation is highly relevant because both of these phenomena constitute crucial processes which critically influence the prognosis of patients. Numerous in vivo and in vitro trials over the past years have fostered a better understanding of the pathophysiology of capillary leak. Furthermore, promising data in animal models show that therapeutic modulation of endothelial barrier function and microcirculation can be achieved by stabilizing endothelial cAMP (cyclic adenosine monophosphate) levels followed by activation of Rho-GTPase Rac1, e. g. by phosphodiesterase 4 inhibitors. This review summarizes and discusses recent findings of cellular mechanisms and in vivo trials.

Effects of medical causes, role concepts and treatment stages on quality of life in involuntary childless men.

Goal of this study was to investigate differences in quality of life in men contingent upon various fertility treatment stages, infertility causes and adoption of roles. A quantitative study with n = 115 men in three German fertility centres was devised. Participants completed a standardised, fertility-specific questionnaire devised for men (TLMK), sociodemographic and role items. Men having experienced severe medical conditions, for example cancer, reported significant higher quality of life compared to men with other infertility reasons [F(1,56) = 12.77, P = 0.001]. Furthermore, allocating participants into distinctive groups by means of kind and duration of treatment revealed significant group differences [F(2,111) = 4.94, P = 0.009], with quality of life decreasing with the use of more invasive fertility methods. A higher satisfaction with life was also stated by men adopting many tasks in the treatment process. The high quality of life displayed by men having experienced severe medical conditions contains valuable and far-reaching information about possible resilience factors that need to be researched more in detail. The finding of decreasing quality of life in men with the use of more invasive methods in treatment applies for increased psychosocial services in fertility clinics.

Effect of mobile ions on the electric field needed to orient charged diblock copolymer thin films.

We examine the behavior of lamellar phases of charged/neutral diblock copolymer thin films containing mobile ions in the presence of an external electric field. We employ self-consistent field theory and focus on the aligning effect of the electric field on the lamellae. Of particular interest are the effects of the mobile ions on the critical field, the value required to reorient the lamellae from the parallel configuration favored by the surface interaction to the perpendicular orientation favored by the field. We find that the critical field depends strongly on whether the neutral or charged species is favored by the substrates. In the case in which the neutral species is favored, the addition of charges decreases the critical electric field significantly. The effect is greater when the mobile ions are confined to the charged lamellae. In contrast, when the charged species is favored by the substrate, the addition of mobile ions stabilizes the parallel configuration and thus results in an increase in the critical electric field. The presence of ions in the system introduces a new mixed phase in addition to those reported previously.

Improving ergonomics in milking parlors: empirical findings for optimal working heights in five milking parlor types.

Milking postures have shifted from seated milking in tethered stalls to milking in a standing position in parlors. However, the musculoskeletal workload of dairy farmers remains high. Previous studies have shown that different working heights affect ergonomics, but they could not objectively evaluate and quantify the workload. The aim of the present study was to assess the effect of working height in different milking parlor types on the milker's workload during the task of attaching milking clusters. Computer-assisted recording and long-term analysis of movements were used to record positions of joints and body regions while performing certain tasks in terms of angular degrees of joints (ADJ) according to the neutral zero method. The 5th, 50th, and 95th percentiles described the distribution of angular degree values measured for each joint. The ADJ were evaluated according to international standards and other scientific literature on the issue to assess the muscular load. The workload was compared between 5 parlor types (auto tandem, herringbone 30°, herringbone 50°, parallel, and rotary) on 15 farms with 2 subjects per parlor and 1 milking period per subject. The working height was defined as a coefficient based on the milker's body height, the floor level, and the cow's udder height. The data recorded during the attachment task were analyzed using generalized linear mixed-effects models taking into account the hierarchical experimental design. The results indicated that the interaction of the cow's udder height, the milker's body height, and the parlor type had a larger effect on ergonomics than each parameter had independently. The interaction was significant in at least 1 of the 3 percentiles in 28 out of 31 ADJ. The postural differences between parlor types, however, were minor. A milking health formula was created to calculate the ideal depth of pit by considering the parlor type, the milker's height, and the mean herd udder height. This formula can be used to develop individual recommendations for future parlor construction.

How cholesterol could be drawn to the cytoplasmic leaf of the plasma membrane by phosphatidylethanolamine.

In the mammalian plasma membrane, cholesterol can translocate rapidly between the exoplasmic and cytoplasmic leaves, so that its distribution between them should be given by the equality of its chemical potential in the leaves. Due to its favorable interaction with sphingomyelin, which is almost entirely in the outer leaf, one expects the great majority of cholesterol to be there also. Experimental results do not support this, implying that there is some mechanism attracting cholesterol to the inner leaf. We hypothesize that it is drawn there to reduce the bending free energy of the membrane caused by the presence of PE (phosphatidylethanolamine). It does this in two ways: first by simply diluting the amount of PE in the inner leaf, and second by ordering the tails of the PE to reduce its spontaneous curvature. Incorporating this mechanism into a model free energy for the bilayer, we find that between 50 and 60% of the total cholesterol should be in the inner leaf of human erythrocytes.

Macroscopic phase separation, modulated phases, and microemulsions: a unified picture of rafts.

We simulate a simple phenomenological model describing phase behavior in a multicomponent membrane, a model capable of producing macroscopic phase separation, modulated phases, and microemulsions, all of which have been discussed in terms of raft phenomena. We show that one effect of thermal fluctuations on the mean-field phase diagram is that it permits a direct transition between either one of the coexisting liquid phases to a microemulsion. This implies that one system exhibiting phase separation can be related to a similar system exhibiting the heterogeneities characteristic of a microemulsion. The two systems could differ in their average membrane composition or in the relative compositions of their exoplasmic and cytoplasmic leaves. The model provides a unified description of these raft-associated phenomena.

[Vibration as a risk of mastitis during milking]. / Vibration als Mastitis-Risiko beim Melken.

INTRODUCTION: Mastitis is one of the most important factor diseases in dairy cattle worldwide. Milking technique represents one of the factors involved in the development of mastitis. The purpose of this study was to investigate the influence of vibrations during milking on the rate of clinical and subclinical mastitis. For this purpose, milking measurements, tank milk analyses and survey forms (general farm data, assessment of milking work and milking hygiene, teat condition, feeding, farm problems, animal behavior) were assessed in 8 Swiss dairy farms. The results show a correlation between present vibrations at the output of the milk meter and increasing bulk milk somatic cell count. Further, a tendency was shown for vibrations at the input of the milk meter to influence bulk milk somatic cell count. Also, a tendency regarding vibrations at the outlet of the milk meter and acute phase protein milk amyloid A was evident. In conclusion, the results suggest that vibration during milking might have a negative effect on udder health. However, further research with a larger number of dairies is needed to make a more generally valid statement.

INTRODUCTION: La mammite est l'une des maladies les plus importantes chez les vaches laitières dans le monde entier. La technique de traite représente l'un des facteurs impliqués dans le développement de la mammite. L'objectif de cette étude était d'étudier l'influence des vibrations pendant la traite sur le taux de mammites cliniques et subcliniques. Pour ce faire, des mesures de traite, des analyses de lait de tank et des formulaires d'enquête (données générales de l'exploitation, évaluation du travail de traite et de l'hygiène de la traite, état des trayons, alimentation, problèmes de l'exploitation, comportement des animaux) ont été évalués dans 8 exploitations laitières suisses. Les résultats montrent une corrélation entre les vibrations présentes à la sortie du compteur à lait et l'augmentation du nombre de cellules somatiques du lait en vrac. En outre, les vibrations à l'entrée du compteur à lait ont tendance à influencer le nombre de cellules somatiques du lait en vrac. De même, une tendance concernant les vibrations à la sortie du compteur à lait et la protéine amyloïde A du lait de phase aiguë a été mise en évidence. En conclusion, les résultats suggèrent que les vibrations pendant la traite pourraient avoir un effet négatif sur la santé de la mamelle. Cependant, des recherches supplémentaires avec un plus grand nombre de laiteries sont nécessaires pour faire une déclaration plus généralement valable.

Model of a raft in both leaves of an asymmetric lipid bilayer.

We present a theory of inhomogeneities in the plasma membrane, or rafts, that can exist in both leaves of the plasma membrane. We note that although neither of the major phospholipid components of the outer leaf, sphingomyelin (SM) nor phosphatidylcholine (PC), evinces a tendency to form phases characterized by nonzero curvature, one of the major components of the inner leaf, phosphatidylethanolamine (PE), displays a strong tendency to do so whereas the other, phosphatidylserine (PS), does not. Therefore, we posit that the concentration difference of PS and PE couples to height fluctuations of the plasma membrane bilayer. This brings about a microemulsion in the inner leaf. Coupling of the concentration difference between PS and PE in the inner leaf and SM and PC in the outer leaf propagates the microemulsion to that leaf as well. The characteristic size of the inhomogeneities is equal to the square-root of the ratio of the bending modulus of the bilayer to its surface tension, a size which is ~100 nm for the plasma membrane. If the coupling between leaves were to be provided by the interchange of cholesterol, then our model raft would consist of SM and cholesterol in the outer leaf and PS and cholesterol in the inner leaf floating in a sea of PC and PE in both leaves.

Hip strength assessment using handheld dynamometry is subject to intertester bias when testers are of different sex and strength.

Handheld dynamometry (HHD) is a promising tool for obtaining reliable hip strength measurements in the clinical setting, but intertester reliability has been questioned, especially in situations where testers exhibit differences in upper-extremity muscle strength (male vs female). The purpose of this study was to examine the intertester reliability concerning strength assessments of hip abduction, adduction, external and internal rotation, flexion and extension using HHD, and to test whether systematic differences in test values exist between testers of different upper-extremity strength. Fifty healthy individuals (29 women), aged 25 ± 5 years were included. Two physiotherapist students (one female, one male) of different upper-extremity strength performed the measurements. The tester order and strength test order were randomized. Intraclass correlation coefficients were used to quantify reliability, and ranged from 0.82 to 0.91 for the six strength test. The female tester systematically measured lower strength values for all isometric strength tests (P < 0.05). In hip strength assessments using HHD, systematic bias exists between testers of different sex, which is likely explained by differences in upper-extremity strength. Hence, to improve intertester reliability, the dynamometer likely needs external fixation, as this will eliminate the influence of differences in upper-extremity strength between testers.

Phase behavior of lipid bilayers under tension.

Given the proposed importance of membrane tension in regulating cellular functions, we explore the effects of a finite surface tension on phase equilibrium using a molecular theory that captures the quantitative structure of the phase diagram of the tensionless DPPC/DOPC/Cholesterol lipid bilayer. We find that an increase in the surface tension decreases the temperature of the transition from liquid to gel in a pure DPPC system by ∼1.0 K/(mN/m), and decreases the liquid-disordered to liquid-ordered transition at constant chemical potentials by approximately the same amount. Our results quantitatively isolate the role of tension in comparison to other thermodynamic factors, such as pressure, in determining the phase behavior of lipid bilayers.

Desflurane-induced and ischaemic postconditioning against myocardial infarction are mediated by Pim-1 kinase.

BACKGROUND: Anaesthetic-induced (APOST) and ischaemic postconditioning (IPOST) against myocardial infarction are mediated via phosphatidylinositol-3-kinase/Akt. Pim-1 kinase is acting downstream of Akt and has recently been demonstrated to enhance cardiomyocyte survival. We tested the hypothesis that both APOST and IPOST are mediated by Pim-1 kinase. METHODS: Pentobarbital-anaesthetized male C57BL/6 mice were subjected to 45-min coronary artery occlusion (CAO) and 3-h reperfusion. Animals received either no intervention, the Pim-1 kinase inhibitor II (10 µg/g intraperitoneally) or its vehicle dimethy sulfoxide (10 µl/g intraperitoneally). Three minutes prior to the end of CAO, 1.0 minimum alveolar concentration desflurane was administered for 18 min alone or in combination with Pim-1 kinase inhibitor II. IPOST was induced by three cycles of each 10-s ischaemia/reperfusion, and animals received either IPOST alone or in combination with Pim-1 kinase inhibitor II. Infarct size was determined with triphenyltetrazolium chloride and area at risk with Evans blue. Protein expression of Pim-1 kinase, Bad, phospho-Bad(Ser112) and B-cell lymphoma 2 was determined using Western immunoblotting analysis. RESULTS: Infarct size in control animals (CON) was 46 ± 3%. Dimethylsulfoxide (47 ± 3%) and Pim-1 kinase inhibitor II (44 ± 5%) did not significantly reduce infarct size. Desflurane (16 ± 2%*; *P < 0.05 vs. CON) and IPOST (21 ± 2%*) significantly reduced infarct size compared with CON. Inhibition of Pim-1 kinase abolished desflurane-induced postconditioning (46 ± 4%) and IPOST (44 ± 5%). Western blot analysis revealed that only desflurane enhances phosphorylation of Bad at serine 112 that was abrogated by Pim-1 kinase inhibitor II. CONCLUSION: These data suggest that Pim-1 kinase mediates both desflurane-induced postconditioning and IPOST in mice.

Effects of oral rehydration and external cooling on physiology, perception, and performance in hot, dry climates.

Only limited research evaluates possible benefits of combined drinking and external cooling (by pouring cold water over the body) during exercise. Therefore, this study examined cold water drinking and external cooling on physiological, perceptual, and performance variables in hot, dry environments. Ten male runners completed four trials of walking 90 min at 30% VO(2max) followed by running a 5-km time trial in 33 ± 1 °C and 30 ± 4% relative humidity. Trials examined no intervention (CON), oral rehydration (OR), external cooling (EC), and oral rehydration plus external cooling (OR + EC). Investigators measured rectal temperature, skin temperatures, heart rate, thirst, thermal sensation, and ratings of perceived exertion (RPE). Oral rehydration (OR and OR + EC) significantly lowered heart rate (P < 0.001) and thirst (P < 0.001) compared with nondrinking (CON and EC) during low-intensity exercise. External cooling (EC and OR + EC) significantly reduced chest and thigh temperature (P < 0.001), thermal sensation (P < 0.001), and RPE (P = 0.041) compared with non-external cooling (CON and OR) during low-intensity exercise. Performance exhibited no differences (CON = 23.86 ± 4.57 min, OR = 22.74 ± 3.20 min, EC = 22.96 ± 3.11 min, OR + EC = 22.64 ± 3.73 min, P = 0.379). Independent of OR, pouring cold water on the body benefited skin temperature, thermal sensation, and RPE during low-intensity exercise in hot, dry conditions but failed to influence high-intensity performance.

'A sorrow shared ': a qualitative content analysis of what couples with recurrent miscarriages expect from one another and their families and friends.

STUDY QUESTION: When couples have to face recurrent pregnancy loss (RPL), what are the partners' wishes and needs and what is their perception of helpful and unhelpful factors with regard to their own, their partners' and their families' and friends' ways of dealing with the problem? SUMMARY ANSWER: Women and men with repeated miscarriages want open communication about their losses, but expect a sensitive and empathetic attitude from others, not pity or trivialization. WHAT IS KNOWN ALREADY: RPL not only causes the women affected and their partners considerable emotional distress; it also has an impact on the couples' relationships and the way they relate to their families and friends. Studies suggest that women have a greater need than their male partners to talk about their losses and that these differences may lead to dissatisfaction and cause relational tension. In addition, men often assume a 'mainstay' role, supporting their partners and displaying fortitude in the face of distress. As yet, however, little research has been conducted so far on the question of what the members of couples with RPL expect from one another and from their families and friends. STUDY DESIGN SIZE DURATION: The study sample consisted of 147 couples and 17 women with at least 2 miscarriages attending the special unit for RPL at the University Women's Hospital in Heidelberg (Germany) for the first time between September 2018 and October 2020 (response rate: 82.7%). The patients were asked to participate in this combined qualitative and questionnaire study. PARTICIPANTS/MATERIALS SETTING METHODS: In order to explore the wishes and needs of those affected in more detail, the free text responses obtained were examined in this study by using qualitative content analysis. Categories and subcategories were created inductively to summarize and systematize content. MAIN RESULTS AND THE ROLE OF CHANCE: Patients affected by RPL want their partners and their families and friends to deal with the topic openly and empathically. In the partnership itself, acceptance of individual grieving modes and sharing a common goal are important factors. Men, in particular, want their partners to be optimistic in facing up to the situation. Regarding communication with family and friends, it transpired that 'good advice', playing the matter down, inquiries about family planning, pity and special treatment are explicitly not appreciated. LIMITATIONS REASONS FOR CAUTION: The sample was a convenience sample, so self-selection effects cannot be excluded. In addition, the level of education in the sample was above average. Accordingly, the sample cannot be regarded as representative. The results of the content analysis are based on the respondents' written answers to open-ended questions in the questionnaire. Unlike qualitative interview studies, further questioning was not possible in the case of ambiguities or to request more details. WIDER IMPLICATIONS OF THE FINDINGS: Frank and sincere communication about miscarriages and about one's own emotions and needs should be promoted both in the partnership and among family members and friends in order to strengthen the potential of social support as a resource. Open communication about the different needs of both partners is necessary to create mutual understanding. The results show the importance not only of empathy and consideration for the couples concerned but also their desire not to be pitied. Striking a fine balance between fellow-feeling and pity may also lead to tension, and this potential dilemma should be addressed in psychosocial counselling. Overall, the study contributes to a better understanding of what couples want from their families and friends when they are attempting to come to terms with RPL and highlights potential challenges in the interaction between affected couples and their families and friends. STUDY FUNDING/COMPETING INTERESTS: No funding was received for this study. None of the authors declared any conflicts of interest. TRIAL REGISTRATION NUMBER: DRKS00014965.