Glutathione treatment protects the rat liver against injury after warm ischemia and Kupffer cell activation

Background/Aim: The generation of reactive oxygen species by activated Kupffer cells (KC) may contribute to reperfusion injury of the liver during liver transplantation or resection. The aim of our present studies was to investigate (1) prevention of hepatic reperfusion injury after warm ischemia by administration of the antioxidant glutathione (GSH) and (2) whether GSH confers protection through influences on KC toxicity. Methods: Isolated perfused rat livers were subjected to 1 h of warm ischemia followed by 90 min of reperfusion without (n = 5) or with GSH or catalase (n = 4-5 each). Selective KC activation by zymosan (150 mug/ml) in continuously perfused rat livers was used to investigate KC-related liver injury. Results: Postischemic infusion of 0.1, 0.5, 1.0 and 2.0 mM GSH, but not 0.05 mM GSH prevented reperfusion injury after warm ischemia as indicated by a marked reduction of sinusoidal LDH efflux by up to 83 +/- 13% (mean +/- SD; p < 0.05) and a concomitant significant improvement of postischemic bile flow by 58 +/- 27% (p < 0.05). A similar protection was conveyed by KC blockade with gadolinium chloride indicating prevention of KC-related reperfusion injury by postischemic GSH treatment. Postischemic treatment with catalase (150 U/ml) resulted in a reduction of LDH efflux by 40 +/- 9% (p < 0.05). Accordingly, catalase as well as GSH (0.1-2.0 mM) nearly completely prevented the increase in LDH efflux following selective :KC activation by zymosan in continously perfused rat livers. Conclusion: Postischemic administration of GSH protects the liver against reperfusion injury after warm ischemia. Detoxification of KC-derived hydrogen peroxide seem to be an important feature of the protective mechanisms. Copyright (C) 2002 S. Karger AG, Basel

TissUExM protocol for ultrastructure expansion microscopy of zebrafish larvae and mouse embryos

Expansion microscopy of millimeter-large mechanically heterogeneous tissues, such as whole vertebrate embryos, has been limited, particularly when combined with post-expansion immunofluorescence. Here, we present a protocol to perform ultrastructure expansion microscopy of whole vertebrate embryos, optimized to perform post-expansion labeling. We describe steps for embedding and denaturing zebrafish larvae or mouse embryos. We then detail procedures for hydrogel handling and mounting. This protocol is particularly well suited for super-resolution imaging of macromolecular protein complexes in situ but does not preserve lipids. For complete details on the use and execution of this protocol, please refer to Steib et al.1

Amiloride reduces portal hypertension in rat liver cirrhosis

Objective This study aimed to investigate the effect of amiloride on portal hypertension. Amiloride is known to inhibit Na(+)/H(+) exchangers on activated hepatic stellate cells. Methods Liver cirrhosis in rats was induced by bile duct ligation (BDL) or thioacetamide (TAA) administration. The effects of zymosan for Kupffer cell (KC) activation or a thromboxane (TX) analogue (U46619) were tested in isolated perfused livers of cirrhotic rats and in vivo. Downstream mechanisms were investigated using Rho kinase inhibitor (Y-27632) or amiloride. Acute and chronic effects of amiloride and canrenoate on portal pressure were compared in perfused livers and in vivo. TXB(2) efflux was measured by ELISA. The phosphorylation state of moesin (p-moesin) as an indicator of Rho kinase activity and expression of the thromboxane synthase were assessed by western blot analyses. The activity of hepatic stellate cells was analysed by western blot and staining for alpha-smooth muscle actin (alpha-SMA). Results In BDL rats, KC activation via zymosan increased portal pressure. This was attenuated by the Rho kinase inhibitor Y-27632. Increased thromboxane efflux following zymosan infusion remained unaltered by Y-27632. The infusion of amiloride attenuated zymosan- and U46619-induced increases in portal perfusion pressure. In vivo, direct administration of amiloride, but not of canrenoate, lowered portal pressure. In TAA and BDL rats, treatment with amiloride for 3 days reduced basal portal pressure and KC-induced increases in portal pressure whereas canrenoate had no effect. In livers of amiloride-treated animals, the phosphorylation state of moesin and the number of alpha-SMA positive cells were reduced. Conclusions Amiloride lowers portal pressure in rat liver cirrhosis by inhibition of intrahepatic vasocontraction. Therefore, patients with cirrhosis and portal hypertension may benefit from amiloride therapy

Feasibility, effectiveness and acceptability of two perturbation-based treadmill training protocols to improve reactive balance in fall-prone older adults (FEATURE): protocol for a pilot randomised controlled trial

\ua9 Author(s) (or their employer(s)) 2023. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.Introduction Perturbation-based balance training (PBT) targets the mechanism of falls (eg, slipping, tripping) to specifically train the recovery actions needed to avoid a fall. This task-specific training has shown great promise as an effective and efficient intervention for fall prevention in older adults. However, knowledge about the dose-response relationship of PBT, as well as its feasibility and acceptability in older adults with increased risk of falling is still limited. Thus, the aim of this study is to compare the effectiveness of two different treadmill PBT protocols for improving reactive balance control in fall-prone older adults, and to evaluate the feasibility and acceptability of these protocols. Methods and analysis The study is designed as a pilot randomised controlled trial with a 6-week intervention and 6-week follow-up period. Thirty-six community-dwelling, fall-prone (Timed Up and Go &gt;12 s, habitual gait speed &lt;1.0 m/s and/or fall history) older adults will be randomised (1:1) to receive six (weeks 1-6) or two treadmill PBT sessions (weeks 1+6) plus four conventional treadmill training sessions (weeks 2-5). Training sessions are conducted 1 7/week for 30 min. Each PBT will include 40 perturbations in anterior-posterior and mediolateral directions. Reactive balance after perturbations in standing (Stepping Threshold Test (STT)) and walking (Dynamic Stepping Threshold Test (DSTT)) will be assessed as the primary outcome for effectiveness. Secondary outcomes are spatiotemporal and kinematic parameters collected during STT, DSTT and PBT, maximum perturbation magnitude for each PBT session, static and dynamic balance, physical capacity, physical activity, concerns with falling and executive functions. Feasibility will be assessed via training adherence, drop-out rate, perturbations actually performed and adverse events; and acceptability via self-designed questionnaire and focus groups. Ethics and dissemination The study has been approved by the Ethics Committee of the Medical Faculty Heidelberg (S-602/2022). Findings will be disseminated through publications in peer-reviewed journals and conference presentations. Trial registration number DRKS00030805

Retrieving and recontextualising VET theory

To what extent can we speak of theory specific to vocational education and training (VET) and what is its relevance today? This Special Issue 19 of bwp@ aims to (re)ignite academic discourse on VET theory, retrieving earlier theorisation specific to this field, mainly from Germany, Austria, and Switzerland (the DACH countries) and connecting it both with international perspectives and contemporary debates. We invite papers in English or German that engage with these debates. From an international perspective, the DACH countries are extraordinary both in their proposition of theories of VET sui generis and especially in their influence on the field of policy and practice. By contrast, for example in English-speaking countries, influential theories that address the question of vocational study, whilst drawing extensively on philosophical and social science concepts, developed largely in opposition to policy and practices that positioned vocational learning as an inferior pathway and narrowed its educational scope. The dual apprenticeship model has been widely imitated internationally but without regard to the social partnerships, labour markets and education workforce developed in Germany, and the theories that shaped this system are neither translated nor widely discussed in other languages. While there are several approaches to VET theory, the core of all these approaches is a framework of normative goals of education, a characterisation of how these goals can be reached through vocational education in particular and the formulation of (education) policy implications that are necessary for successful implementation. For example, Kerschensteiner (1901, 1966/1904) in his emphasis on civic virtues as a central aim of education, drew attention to the possibility of attaining such virtues through work, but acknowledged the necessity of a foundation in general education. In contrast, post-war critical approaches, which regularly draw on critical theory (see Habermas 1968; Horkheimer & Adorno 1947), and can be dated to the 1960s and 1970s, identify autonomy and emancipation as central objectives of education (e.g. Lempert 1971; Blankertz 1974, 1979, 1982). The main challenge of VET theory was to explain how these goals also can be reached through vocational education. Blankertz’s answer lay in emphasizing the role of the VET school in widening and deepening knowledge associated with the workspace. The value of these approaches for contemporary VET is dependent on their adaptation to contemporary problems that VET and society are experiencing: migration and integration; climate change and resource consumption, digitisation and globalisation. The tertiarisation of both the economy and of education, as the service sector employs a greater proportion of the population and a growing number of young people enter higher education, even in Germany, also calls into question the relevance of established VET theories. As the structures and forms of organisation that have sustained VET since the 1970s have given way to new social formations and new forms of precarity, the relevance of theories developed during the long period of post-war growth is called into question. Correspondingly, whether a critical standpoint can still be clearly located after the “fall of metaphysics” (Adorno 1998/1965) has also been deemed as questionable (cf. Schäfer 2005). Furthermore, a succession of de-centring approaches including post-structuralist, postmodernist and post-anthropocentric paradigms has suggested the erosion of earlier ‘grand narratives’, the supersession of the ‘enlightenment project’ and even questioned the progressive potential of human labour that explicitly VET theories tend to take for granted. Call for Papers bwp@ Spezial 19 2 Against this background, the bwp@ Special invites contributions that address both VET theory and its application to contemporary issues. The aim is to stimulate a discussion on the continuing significance of VET theory. The aim is to provide space for ideas on how the discipline and its subject could position itself, both nationally and beyond national borders, in its normative-theoretical contours and/or in relation to VET policy and VET practice

Survivor of a traumatic atlanto-occipital dislocation

AbstractAtlanto-occipital dislocation is a devastating ligamentous injury that most often turns fatal. However, because of on-site resuscitation improvements, the emergency teams are increasingly dealing with this condition. We report a rare case of atlanto-occipital dislocation (AOD) in a surviving patient with more than one-year follow-up. The mechanism of injury appears to be an extreme hyperextension applied to the head. This injury occurs more frequently in children since they are anatomically predisposed (flat articulation between the occiput and the atlas, increased ligamentous laxity). The diagnosis should be suggested by severe neurological injury after high trauma but also post-traumatic cardiorespiratory deficit. There have been reports of atlanto-occipital dilocations without neurologic impairment. A radiographic examination must be performed and lateral cervical radiographs should be acquired. However, additional imaging with CT or MRI may be required to aid diagnosis of AOD in cases in which radiographic findings are equivocal. Once the diagnosis of AOD has been confirmed, an anatomical classification should be made according to the magnitude of displacement. Fatal lesions are of neurological and vascular origin and some authors advocate the systematic use of angiography. Consensus regarding the management of AOD in adults has been achieved. Occipito-cervical arthrodesis is the recommended treatment option. We advocate a two-stage surgery: the patient is initially fitted with a halo vest then occipitocervical fusion is performed. Surgical treatment should be combined with cardiorespiratory management. The emergency teams should get familiar with this injury since they will be increasingly confronted to it. Early recognition and standard appropriate management is essential to avoid delayed treatment and complications

Comments about birefringence dispersion, with group and phase birefringence measurements in polarization-maintaining fibers

A recent JEOS-RP publication proposed Comments about Dispersion of Light Waves, and we present here complementary comments for birefringence dispersion in polarization-maintaining (PM) fibers, and for its measurement techniques based on channeled spectrum analysis. We start by a study of early seminal papers, and we propose additional explanations to get a simpler understanding of the subject. A geometrical construction is described to relate phase birefringence to group birefringence, and it is applied to the measurement of several kinds of PM fibers using stress-induced photo-elasticity, or shape birefringence. These measurements confirm clearly that the difference between group birefringence and phase birefringence is limited to 15–20% in stress-induced PM fibers (bow-tie, panda, or tiger-eye), but that it can get up to a 3-fold factor with an elliptical-core (E-core) fiber. There are also surprising results with solid-core micro-structured PM fibers, that are based on shape birefringence, as E-core fibers

Identification of chlamydomonas central core centriolar proteins reveals a role for human WDR90 in ciliogenesis

Centrioles are evolutionarily conserved macromolecular structures that are fundamental to form cilia, flagella, and centrosomes. Centrioles are 9-fold symmetrical microtubule-based cylindrical barrels comprising three regions that can be clearly distinguished in the Chlamydomonas reinhardtii organelle: an ∼100-nm-long proximal region harboring a cartwheel; an ∼250-nm-long central core region containing a Y-shaped linker; and an ∼150-nm-long distal region ending at the transitional plate. Despite the discovery of many centriolar components, no protein has been localized specifically to the central core region in Chlamydomonas thus far. Here, combining relative quantitative mass spectrometry and super-resolution microscopy on purified Chlamydomonas centrioles, we identified POB15 and POC16 as two proteins of the central core region, the distribution of which correlates with that of tubulin glutamylation. We demonstrated that POB15 is an inner barrel protein within this region. Moreover, we developed an assay to uncover temporal relationships between centriolar proteins during organelle assembly and thus established that POB15 is recruited after the cartwheel protein CrSAS-6 and before tubulin glutamylation takes place. Furthermore, we discovered that two poc16 mutants exhibit flagellar defects, indicating that POC16 is important for flagellum biogenesis. In addition, we discovered that WDR90, the human homolog of POC16, localizes to a region of human centrioles that we propose is analogous to the central core of Chlamydomonas centrioles. Moreover, we demonstrate that WDR90 is required for ciliogenesis, echoing the findings in Chlamydomonas. Overall, our work provides novel insights into the identity and function of centriolar central core components

WDR90 is a centriolar microtubule wall protein important for centriole architecture integrity

Centrioles are characterized by a nine-fold arrangement of microtubule triplets held together by an inner protein scaffold. These structurally robust organelles experience strenuous cellular processes such as cell division or ciliary beating while performing their function. However, the molecular mechanisms underlying the stability of microtubule triplets, as well as centriole architectural integrity remain poorly understood. Here, using ultrastructure expansion microscopy for nanoscale protein mapping, we reveal that POC16 and its human homolog WDR90 are components of the microtubule wall along the central core region of the centriole. We further found that WDR90 is an evolutionary microtubule associated protein. Finally, we demonstrate that WDR90 depletion impairs the localization of inner scaffold components, leading to centriole structural abnormalities in human cells. Altogether, this work highlights that WDR90 is an evolutionary conserved molecular player participating in centriole architecture integrity.</jats:p