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The oncogene DJ-1 has been originally identified as a suppressor of PTEN. Further on, loss-of-function mutations have been described as a causative factor in Parkinson's disease (PD). DJ-1 has an important function in cellular antioxidant responses, but its role in central metabolism of neurons is still elusive. We applied stable isotope assisted metabolic profiling to investigate the effect of a functional loss of DJ-1 and show that DJ-1 deficient neuronal cells exhibit decreased glutamine influx and reduced serine biosynthesis. By providing precursors for GSH synthesis, these two metabolic pathways are important contributors to cellular antioxidant response. Down-regulation of these pathways, as a result of loss of DJ-1 leads to an impaired antioxidant response. Furthermore, DJ-1 deficient mouse microglia showed a weak but constitutive pro-inflammatory activation. The combined effects of altered central metabolism and constitutive activation of glia cells raise the susceptibility of dopaminergic neurons towards degeneration in patients harboring mutated DJ-1. Our work reveals metabolic alterations leading to increased cellular instability and identifies potential new intervention points that can further be studied in the light of novel translational medicine approaches.

Introduction: The Earth experienced three major extinctions during the Paleozoic Era, usually related to rapid and sudden climate changes that were caused by one or both internal and external events such as plate tectonics, large volcanic eruptions, changes in the Earth's orbit, and/or asteroid impacts [1]. Indeed, the Devonian-Carboniferous boundary is characterized by a record of widespread oceanic anoxia, which coincided with massive marine biodiversity losses. Astronomical insolation changes, linked to Earth's eccentricity, may have played a role in the increase of ocean anoxia and other environmental perturbations [2]. The flux of extraterrestrial material to the Earth does not appear to have been constant over time. Discrete events in the solar system may have led to increases in the flux of asteroids, meteorites, and micrometeorites with time. While most of this material weathers away quickly under the oxidizing conditions at the Earth surface, some refractory minerals or fossil micrometeorites may resist alteration and diagenesis [3,4]. To date, various increases in the extraterrestrial influx have been identified in the geological record, based on enrichments in spinel group minerals but also I-type micrometeorites in various types of sediments [5]. However, none of these higher extraterrestrial fluxes has convincingly been linked to a mass extinction event, or changes in the eccentricity of the Earth's orbit. Schmitz et al. [6] worked on the sedimentary records at the Frasnian-Famennian boundary, which marks another Late Devonian extinction (the Upper Kellwasser Event), searching for an increase in the extraterrestrial materials, looking for Cr-spinel as a proxy using heavy acid dissolution of the rocks. They did not find clear evidence for an increased flux of extraterrestrial Cr-spinel crystals, potentially due to insufficiently high amounts of those minerals in the rock. However, Cr-spinels are not found in all types of extraterrestrial materials, and heavy acid extraction methods could have dissolved any remaining extraterrestrial silicate-rich material, which represents an important fraction of the extraterrestrial flux. In addition, Suttle and Genge [7] reported the discovery of fossil micrometeorites, as pristine silicates, iron-type spherules and magnetite or Fe-silicate pseudomorphic spherules, from Late Cretaceous chalk. They investigated both acid dissolution and mechanical separation and discovered different results, both in abundance and type of extraterrestrial materials depending on the extraction methods. In this study, we will compare different methods to extract fossil micrometeorites and other extraterrestrial materials from different lithologies, in order to evaluate the most effective method for the extraction of extraterrestrial materials, while trying to limit any procedural biases. Methods: We collected rocks (carbonates, limestones, and shales) from around the Devonian-Carboniferous boundary that records the Hangenberg extinction event. We plan to apply various extraction methods of extraterrestrial materials on the different lithologies: (i) HCl dissolution, (ii) consecutive HCl and HF dissolution, (iii) consecutive HCl, HF and H2SO4, (iv) surfactant Rewoquat following Jarochowska et al. method [8], (v) heavy liquids separation, and (vi) mechanical disaggregation (crushing). We will examine the residue of those methods and look for extraterrestrial materials, including Cr-spinel crystals as well as whole micrometeorites (silicates, glassy, oxides and irontypes), using optical microscopy, scanning electron microscope (SEM) and micro-X-ray fluorescence (XRF). Discussion: We will first discuss the comparison between the efficiency of the different acids processing, the Rewoquat processing, the heavy liquids processing and the manual separation, with the various difficulties caused by the rock compositions, and the different methodological biases arising from the differences between extraction procedures. We propose the most effective extraction methods to extract the various types of extraterrestrial material (from silicate to iron-type micrometeorites) to assess the global extraterrestrial flux, and compare our results to more conventional proxies, such as the iridium and helium-3 contents of the bulk rocks, to trace past variations in the extraterrestrial fluxes to the Earth. We also aim to determine the best extraction methods to preserve the oxidized iron-type spherules, as their oxygen isotopic compositions may contain information on their oxidation during atmospheric entry during the late Devonian. By comparison with other time intervals as well as current Antarctic and urban micrometeorite collections we aim to build a framework that will allow us to evaluate fluctuations in the extraterrestrial material influx on Earth, as well as any effects from preservation biases. [ABSTRACT FROM AUTHOR]

Introduction: Nakhlites are a group of basaltic meteorites originating from the mid-Amazonian igneous crust of Mars [1]. They represent an interesting set of samples to study the aqueous alteration at the Martian subsurface, as they contain aqueous alteration phases in olivine called "iddingsite" (e.g. [2]). Studying iddingsite in nakhlites can reveal the water-crust interactions and the hydrothermal physico-chemical conditions that prevailed at the Martian subsurface, with implications for the past habitability of Mars, and in particular for microbial life during the Amazonian period. We studied iddingsite in Caleta el Cobre (CeC) 022, a nakhlite that contains a high amount of alteration products, and compared this meteorite to 7 other nakhlites. Methods: A serie of analyses were conducted on eight nakhlites polished sections: CeC 022, Governador Valadares, Lafayette, Miller Range (MIL) 03346, Nakhla, North-West Africa (NWA) 817, NWA 998 and Yamato (Y) 000953 to assess the mineralogy, the chemical and the isotopic composition of the alteration products in nakhlites: electronic microscopy (FEG-SEM and TEM), EDX microanalyses and chemical maps, LA-ICPMS major elements analyses, Raman spectra, and finally, hydrogen isotopic measurements by SIMS. Results: In all nakhlites, the alteration veins show at least two types of iddingsite (Fig. 1A): (i) a coarse iddingsite in contact with olivine at the border of the alteration veins, showing 50-200 nm crystals, composed of a mixture of 1:1 phyllosilicates of greenalite-cronstedtite composition, and Fe-oxyhydroxides; (ii) a fine iddingsite, in the inner part of the alteration veins, with <10 nm crystals, with a composition close to saponite (2:1 phyllosilicate). Complex chemical zoning of Mg, Ca, Mn, S, P and Al, has been observed on EDS-TEM mappings in NWA 10153. In most nakhlites, olivine grains also display planes of secondary inclusions, composed of pyroxene, magnetite and a void potentially filled by a fluid (Fig. 1B). In most nakhlites, magnetite-pyroxene symplectites are found in olivine grains, often at the border of the secondary inclusion planes. Those secondary inclusions and symplectites can also be observed at the center of the iddingsite veins (Fig. 1A). Also, sulfide-magnetite veinlets are observed at the center of the iddingsite veins, and also crosscutting olivine and pyroxene grains and mesostasis. Finally, organic matter is observed on Raman spectra in the iddingsite of many nakhlites, and is located in coarse iddingsite, as shown by TEM observations of NWA 10153. Discussion and conclusion: Our favored scenario is that the secondary inclusions and the symplectites are formed by a first fluid alteration event triggered by late magmatic fluids, prior to the iddingsite formation event,. These secondary inclusions represent weakness planes that facilitate the circulation of the alteration fluid forming the iddingsite inside the olivine grains. The composition and texture of both types of iddingsite is suggestive of a crystallization by filling of existing fractures, with an alteration fluid enriched in elements from basaltic glass and host olivine dissolution, and changes in the alteration conditions or fluid composition. The sulfide-magnetite veinlets represent either the late stage of the same alteration event as the iddingsite formation, or a different later fluid injection. With the two types of iddingsite, the complex centers of veins and the chemical zoning in the fine iddingsite, we suggest that fluid alteration in the nakhlites has a complex multistage history of fluid injections. [ABSTRACT FROM AUTHOR]

List of abstracts and authors: 1. Bipolar disorder not otherwise specified -overdiagnosed or underdiagnosed? E Allers 2. The prognosis of major depression untreated and treated: Does the data reflect the true picture of the prognosis of this very common disorder? E Allers 3. Can we prolong our patients' life expectancy? Providing a better quality of life for patients with severe mental illness O A Betencourt 4. The scope of ECT practice in South Africa J Benson-Martin, P Milligan 5. Biomarkers for schizophrenia: Can we evolve like cancer therapeutics? P Buckley 6. Relapse in schizophrenis: Major challenges in prediction and prevention P Buckley 7. Informed consent in biological treatments: The right to know the duty to inform I Chetty 8. Effectiveness of a long-acting injectable antipsychotic plus an assertive monitoring programme in first-episode schizophrenia B Chiliza, L Asmal, O Esan, A Ojagbemi, O Gureje, R Emsley 9. Name, shame, fame P Cilliers 10. Can we manage the increasing incidence of violent raging children? We have to! H Clark 11. Serotonin, depression and antidepressant action P Cowen 12. Prevalence and correlates of comorbid psychiatris illness in patients with heroin use disorder admitted to Stikland Opioid Detoxification Unit L Dannatt, K J Cloete, M Kidd, L Weich 13. Investigating the association between diabetes mellitus, depression and psychological distress in a cohort of South African teachers A K Domingo, S Seedat, T M Esterhuizen, C Laurence, J Volmink, L Asmal 14. Neuropeptide S -emerging evidence for a role in anxiety K Domschke 15. Pathogenetics of anxiety K Domschke 16. The effects of HIV on the fronto-striatal system S du Plessis, M Vink, J Joska, E Koutsilieri, C Scheller, B Spottiswoode, D Stein, R Emsley 17. Effects of acute antipsychotic treatment on brain morphology in schizophrenia R Emsley, L Asmal, B Chiliza, S du Plessis, J Carr, A Goosen, M Kidd, M Vink, R Kahn 18. Development of a genetic database resource for monitoring of breast cancer patients at risk of physical and psychological complications K Grant, F J Cronje, K Botha, J P Apffelstaedt, M J Kotze 19. Unipolar mania reconsidered: Evidence from a South African study C Grobler 20. Antipsychotic-induced movement disorders: Occurence and management P Haddad 21. The place of observational studies in assessing the effectiveness of long-acting injectable antipsychotics P Haddad 22. Molecular mechanisms of d-cycloserine in fear extinction: Insights from RNS sequencing S Hemmings, S Malan-Muller, L Fairbairn, M Jalali, E J Oakeley, J Gamieldien, M Kidd, S Seedat 23. Schizophrenia: The role of inflammation DC Henderson 24. Addictions: Emergent trends and innovations V Hitzeroth 25. The socio-cultural-religious context of biological psychiatric practice B Janse van Rensburg 26. Biochemical markers for identifying risk factors for disability progression in multiple sclerosis S Janse van Rensburg, M J Kotze, F J Cronje, W Davis, K Moremi, M Jalali Sefid Dashti, J Gamieldien, D Geiger, M Rensburg, R van Toorn, M J de Klerk, G M Hon, T Matsha, S Hassan, R T Erasmus 27. Alcohol-induced psychotic disorder: Brain perfusion and psychopathology - before and after antipsychotic treatment G Jordaan, J M Warwick, D G Nel, R Hewlett, R Emsley 28.'Pump and dump': Harm reduction strategies for breastfeeding while using substances L Kramer 29. Adolescent neuropsychiatry - an emerging field in South African adolescent psychiatric services A Lachman 30. Recovery versus remission, or what it means to be healthy for a psychiatric patient? B Latecki 31. Holistic methods utilised to normalise behaviours in youth diagnosed with neuro-biochemical disorders P Macqueen 32. Candidate genes and novel polymorphisms for anxiety disorder in a South African cohort N McGregor, J Dimatelis, S M J Hemmings, C J Kinnear, D Stein, V Russel, C Lochner 33. Higher visual functioning A Moodley 34. The effects of prenatal methylmercury exposure on trace element and antioxidant levels in rat offspring following 6-hydroxydopamine-induced neuronal insult Z M Moosa, W M U Daniels, M V Mabandla 35. Paediatric neuropsychiatric movement disorders L Mubaiwa 36. The South African national female offenders study M Nagdee, L Artz, C de Clercq, P de Wet, H Erlacher, S Kaliski, C Kotze, L Kowalski, J Naidoo, S Naidoo, J Pretorius, M Roffey, F Sokudela, U Subramaney 37. Neurobiological consequences of child abuse C Nemeroff 38. What do Stellenbosch Unviversity medical students think about psychiatry - and why should we care? G Nortje, S Suliman, K Seed, G Lydall, S Seedat 39. Neurological soft skins in Nigerian Africans with first episode schizophrenia: Factor structure and clinical correlates A Ojagbemi, O Esan, O Gureje, R Emsley 40. Should psychiatric patients know their MTHFR status? E Peter 41. Clinical and functional outcome of treatment refractory first-episode schizophrenia L Phahladira, R Emsley, L Asmal, B Chiliza 42. Bioethics by case discussion W Pienaar 43. Reviewing our social contract pertaining to psychiatric research in children, research in developing countries and distributive justice in pharmacy W Pienaar 44. The performance of the MMSE in a heterogenous elderly South African population S Ramlall, J Chipps, A I Bhigjee, B J Pillay 45. Biological basis addiction (alocohol and drug addiction) S Rataemane 46. Volumetric brain changes in prenatal methamphetamine-exposed children compared with healthy unexposed controls A Roos, K Donald, G Jones, D J Stein 47. Single voxel proton magnetic resonance spectroscopy of the amygdala in social anxiety disorder in the context of early developmental trauma D Rosenstein, A Hess, S Seedat, E Meintjies 48. Discussion of HDAC inhibitors, with specific reference to supliride and its use during breastfeeding J Roux 49. Prevalence and clinical correlates of police contact prior to a first diagnosis of schizophrenia C Schumann, L Asmal, K Cloete, B Chiliza, R Emsley 50. Are dreams meaningless? M Solms 51. The conscious id M Solms 52. Depression and resilience in HIV-infected women with early life stress: Does trauma play a mediating role? G Spies, S Seedat 53. State of affairs analysis for forensic psychiatry in SA U Subramaney 54. Escitalopram in the prevention of post-traumatic stress disorder: A pilot randomised controlled trial S Suliman, S Seedat, J Pingo, T Sutherland, J Zohar, D J Stein 55. Epigenetic consequences of adverse early social experiences in primates S Suomi 56. Risk, resilience, and gene x environment interactions in primates S Suomi 57. Biological aspects of anorexia nervosa C Szabo 58. Agents used and profiles of non-fatal suicidal behaviour in East London H Uys 59. The contributions of G-protein coupled receptor signalling to opioid dependence J van Tonder 60. Emerging trend and innovation in PTSD and OCD J Zohar 61. Making the SASOP treatment guidelines operational E Allers Poster Presentations 62. Neuropsychological deficits in social anxiety disorder in the context of early developmental trauma S Bakelaar, D Rosenstein, S Seedat 63.Social anxiety disorder in patients with or without early childhood trauma: Relationship to behavioral inhibition and activation and quality of life S Bakelaar, C Bruijnen, A Sambeth, S Seedat 64. Exploring altered affective processing in obssessive compulsive disorder symptom subtypes E Breet, J Ipser, D Stein, C Lochner 65. To investigate the bias toward recognising the facial expression of disgust in obsessive compulsive disorder as well as the effect of escitalopram E Breet, J Ipser, D Stein, C Lochner 66. A fatal-case of nevirapine-induced Stevens-Johnson's syndrome in HIV mania A Bronkhorst, Z Zingela, W M Qwesha, B P Magigaba 67. Association of the COMT G472A (met/met) genotype with lower disability in people diagnosed with multiple sclerosis W Davis, S J van Rensburg, L Fisher, F J Cronje, D Geiger, M J Kotze 68. Homocycsteine levels are associated with the fat mass and obesity associated gene FTO(intron 1 T>A) polymorphism in MS patients W Davis, S J Van Rensburg, M J Kotze, L Fisher, M Jalali, F J Cronje, K Moremi, J Gamieldien, D Geiger, M Rensburg, R van Toorn, M J de Klerk, G M Hon, T Matsha, S Hassan, R T Erasmus 69. Analysis of the COMT 472 G>A (rs4680) polymorphism in relation to environmental influences as contributing factors in patients with schizophrenia D de Klerk, S J van Rensburg, R A Emsley, D Geiger, M Rensburg, R T Erasmus, M J Kotze 70. Dietary folate intake, homocysteine levels and MTHFR mutation detection in South African patients with depression: Test development for clinical application D Delport, N vand der Merwe, R Schoeman, M J Kotze 71. The use ofexome sequencing for antipsychotic pharmacogenomic applications in South African schizophrenia patients B Drogmoller, D Niehaus, G Wright, B Chiliza, L Asmal, R Emsley, L Warnich 72. The effects of HIV on the ventral-striatal reward system S du Plessis, M Vink, J Joska, E Koutsilieri, C Scheller, B Spottiswoode, D Stein, R Emsley 73. Xenomelia relates to asymmetrical insular activity: A case study of fMRI S du Plessis, M Vink, L Asmal 74. Maternal mental helath: A prospective naturalistic study of the outcome of pregancy in women with major psychiatric disorders in an African country E du Toit, L Koen, D Niehaus, B Vythilingum, E Jordaan, J Leppanen 75. Prefrontal cortical thinning and subcortical volume decrease in HIV-positive children with encephalopathy J P Fouche, B Spottiswoode, K Donald, D Stein, J Hoare 76. H-magnetic resonance spectroscopy metabolites in schizophrenia F Howells, J Hsieh, H Temmingh, D J Stein 77. Hypothesis for the development of persistent methamphetamine-induced psychosis J Hsieh, D J Stein, F M Howells 78. Culture, religion, spirituality and psychiatric practice: The SASOP Spirituality and Psychiatry Special Interest Group Action Plan for 2012-2014 B Janse van Rensburg 79. Cocaine reduces the efficiency of dopamine uptake in a rodent model of attention-deficit/hyperactivity disorder: An in vivo electrochemical study L Kellaway, J S Womersley, D J Stein, G A Gerhardt, V A Russell 80. Kleine-Levin syndrome: Case in an adolescent psychiatric unit A Lachman 81. Increased inflammatory stress specific clinical, lifestyle and therapeutic variables in patients receiving treatment for stress, anxiety or depressive symptoms H Luckhoff, M Kotze, S Janse van Rensburg, D Geiger 82. Catatonia: An eight-case series report M Mabenge, Z Zingela, S van Wyk 83. Relationship between anxiety sensitivity and childhood trauma in a random sample of adolescents from secondary schools in Cape Town L Martin, M Viljoen, S Seedat 84. 'Making ethics real'. An overview of an ethics course presented by Fraser Health Ethics Services, BC, Canada JJ McCallaghan 85. Clozapine discontinuation rates in a public healthcare setting M Moolman, W Esterhuysen, R Joubert, J C Lamprecht, M S Lubbe 86. Retrospective review of clozapine monitoring in a publica sector psychiatric hospital and associated clinics M Moolman, W Esterhuysen, R Joubert, J C Lamprecht, M S Lubbe 87. Association of an iron-related TMPRSS6 genetic variant c.2007 C>7 (rs855791) with functional iron deficiency and its effect on multiple sclerosis risk in the South African population K Moremi, S J van Rensburg, L R Fisher, W Davis, F J Cronje, M Jalali Sefid Dashti, J Gamieldien, D Geiger, M Rensburg, R van Toorn, M J de Klerk, G M Hon, T Matsha, S Hassan, R T Erasmus, M Kidd, M J Kotze 88. Identifying molecular mechanisms of apormophine-induced addictive behaviours Z Ndlazi, W Daniels, M Mabandla 89. Effects of lifestyle factors and biochemistry on the major neck blood vessels in patients with mutiple sclerosis M Nelson, S J van Rensburg, M J Kotze, F Isaacs, S Hassan 90. Nicotine protects against dopamine neurodegenration and improves motor deficits in a Parkinsonian rat model N Ngema, P Ngema, M Mabandla, W Daniels 91. Cognition: Probing anatomical substrates H Nowbath 92. Chronic exposure to light reverses the effects of maternal separation on the rat prefrontal cortex V Russel, J Dimatelis 93. Evaluating a new drug to combat Alzheimer's disease S Sibiya, W M U Daniels, M V Mabandla 94. Structural brain changes in HIV-infected women with and without childhood trauma G Spies, F Ahmed, C Fennema-Notestine, S Archibald, S Seedat 95. Nicotine-stimulated release of hippocampal norepinephrine is reduced in an animal model of attention-deficit/ hyperactivity disorder: the spontaneously hypertensive rat T Sterley 96. Brain-derive neurotrophic factor (BDNF) protein levels in anxiety disorders: Systematic review and meta-regression analysis S Suliman, S M J Hemmings, S Seedat 97. A 12-month retrospective audit of the demographic and clinical profile of mental healthcare users admitted to a district level hospital in the Western Cape, South Africa E Thomas, K J Cloete, M Kidd, H Lategan 98. Magnesium recurarization: A comparison between reversal of neuromuscular block with sugammadex v. neostigmine/ glycopyrrolate in an in vivo rat model M van den Berg, M F M James, L A Kellaway 99. Identification of breast cancer patients at increased risk of 'chemobrain': Case study and review of the literature N van der Merwe, R Pienaar, S J van Rensburg, J Bezuidenhout, M J Kotze 100. The protective role of HAART and NAZA in HIV Tat protein-induced hippocampal cell death S Zulu, W M U Daniels, M V Mabandla

Conjoined twins are rare variants of monozygotic twins. There are many types of conjoined twins, and they are usually classified at the point at which they are fused. Our patient fitted into a thoraco-omphalopagus type, where there was fusion of the thorax as well as mid-trunk [3]. The most common type of renal fusion anomaly is the horseshoe kidney. It consists of 2 distinct functioning kidneys on each side of the midline, and they are more likely to be connected at the lower poles [4]. We describe here a 30 year-old patient who presented to the antenatal clinic of Chris Hani Baragwanath hospital in Soweto, South Africa at 34+ weeks gestation. Foetal magnetic resonance imaging revealed thoraco-omphalopagus conjoined twins with complex thoracic and abdominal structures, including a horseshoe kidney that was confirmed on post-natal computed tomography.

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An on-chip asymmetric directional coupler (DC) can convert fundamental modes to higher-order modes and is one of the core components of mode-division multiplexing (MDM) technology. In this study, we propose that waveguides of the asymmetric DC can be trimmed by silicon ion implantation to tune the effective refractive index and facilitate mode conversion into higher-order modes. Through this method of tuning, transmission changes of up to 18 dB have been realized with one ion implantation step. In addition, adjusting the position of the ion implantation on the waveguide can provide a further degree of control over the transmission into the resulting mode. The results of this work present a promising new route for the development of high-efficiency, low-loss mode converters for integrated photonic platforms, and aim to facilitate the application of MDM technology in emerging photonic neuromorphic computing.

Micrometeorites are estimated to represent the main part of the present flux of extraterrestrial matter found on the Earth's surface and provide valuable samples to probe the interplanetary medium. Here, we describe large and representative collections of micrometeorites currently available to the scientific community. These include Antarctic collections from surface ice and snow, as well as glacial sediments from the eroded top of nunataks-summits outcropping from the icesheet-and moraines. Collections extracted from deep-sea sediments (DSS) produced a large number of micrometeorites, in particular, iron-rich cosmic spherules that are rarer in other collections. Collections from the old and stable surface of the Atacama Desert show that finding large numbers of micrometeorites is not restricted to polar regions or DSS. The advent of rooftop collections marks an important step into involving citizen science in the study of micrometeorites, as well as providing potential sampling locations over all latitudes to explore the modern flux. We explore their strengths of the collections to address specific scientific questions and their potential weaknesses. The future of micrometeorite research will involve the finding of large fossil micrometeorite collections and benefit from recent advances in sampling cosmic dust directly from the air. This article is part of the theme issue 'Dust in the Solar System and beyond'.

Efficient fiber-to-chip couplers for multi-port access to photonic integrated circuits are paramount for a broad class of applications, ranging, e.g., from telecommunication to photonic computing and quantum technologies. Grating-based approaches are often desirable for providing out-of-plane access to the photonic circuits. However, on photonic platforms characterized by a refractive index ≃ 2 at telecom wavelength, such as silicon nitride or thin-film lithium niobate, the limited scattering strength has thus far hindered the achievement of coupling efficiencies comparable to the ones attainable in silicon photonics. Here we present a flexible strategy for the realization of highly efficient grating couplers on such low-index photonic platforms. To simultaneously reach a high scattering efficiency and a near-unitary modal overlap with optical fibers, we make use of self-imaging gratings designed with a negative diffraction angle. To ensure high directionality of the diffracted light, we take advantage of a metal back-reflector patterned underneath the grating structure by cryogenic deep reactive ion etching of the silicon handle. Using silicon nitride as a testbed material, we experimentally demonstrate coupling efficiency up to - 0.55 dB in the telecom C-band with high chip-scale device yield., (© 2024. The Author(s).)

Almost all mitochondrial proteins are synthesized in the cytosol and subsequently targeted to mitochondria. The accumulation of nonimported precursor proteins occurring upon mitochondrial dysfunction can challenge cellular protein homeostasis. Here we show that blocking protein translocation into mitochondria results in the accumulation of mitochondrial membrane proteins at the endoplasmic reticulum, thereby triggering the unfolded protein response (UPR ER ). Moreover, we find that mitochondrial membrane proteins are also routed to the ER under physiological conditions. The level of ER-resident mitochondrial precursors is enhanced by import defects as well as metabolic stimuli that increase the expression of mitochondrial proteins. Under such conditions, the UPR ER is crucial to maintain protein homeostasis and cellular fitness. We propose the ER serves as a physiological buffer zone for those mitochondrial precursors that cannot be immediately imported into mitochondria while engaging the UPR ER to adjust the ER proteostasis capacity to the extent of precursor accumulation.

Background: Upper gastrointestinal bleeding (UGIB) from malignancies is associated with a poor outcome. Only a small number of studies on gastrointestinal tumor bleeding have been published so far, focusing mainly on bleeding from gastric cancer. Since the information on patients with UGIB from esophageal cancer appears insufficient, this study aimed to present clinical and endoscopic findings, treatment options as well as clinical outcomes such as rebleeding and survival of those patients., Methods: This retrospective analysis included all patients admitted with UGIB from esophageal cancer at our university hospital during a 10-year period., Results: 45 patients were analyzed of whom 26 (57.8%) already had cancer stage IV at index bleeding. 22 (48.9%) patients presented with hemodynamic instability and 30 (66.7%) patients received blood transfusions. Active bleeding was present in 24 (53.3%) patients, of whom 20 (83.3%) received endoscopic therapy. Successful hemostasis was achieved in 18 (90%) of 20 patients with Argon plasma coagulation used most frequently (52.4%). Early and delayed rebleeding occurred in 5 (12.5%) and 11 (27.5%) of all inoperable patients, respectively. Intake of anticoagulation or anti-platelet drugs were risk factors for delayed rebleeding and the median overall survival after index bleeding was 1.2 months., Conclusion: UGIB from esophageal cancer occurred most frequently in advanced tumor stages and was associated with significant blood loss. Even though initial endoscopic therapy was effective, rebleeding occurred in a significant number of patients. Those taking anticoagulants or anti-platelet drugs should be closely monitored for rebleeding. The overall survival after index bleeding was poor.

RNA-sequencing has been proposed as a valuable technique to develop individualized therapy concepts for cancer patients based on their tumor-specific mutational profiles. Here, we aimed to identify drugs and inhibitors in an individualized therapy-based drug repurposing approach focusing on missense mutations for 35 biopsies of cancer patients. The missense mutations belonged to 9 categories (ABC transporter, apoptosis, angiogenesis, cell cycle, DNA damage, kinase, protease, transcription factor, tumor suppressor). The highest percentages of missense mutations were observed in transcription factor genes. The mutational profiles of all 35 tumors were subjected to hierarchical heatmap clustering. All 7 leukemia biopsies clustered together and were separated from solid tumors. Based on these individual mutation profiles, two strategies for the identification of possible drug candidates were applied: Firstly, virtual screening of FDA-approved drugs based on the protein structures carrying particular missense mutations. Secondly, we mined the Drug Gene Interaction (DGI) database (https://www.dgidb.org/) to identify approved or experimental inhibitors for missense mutated proteins in our dataset of 35 tumors. In conclusion, our approach based on virtual drug screening of FDA-approved drugs and DGI-based inhibitor selection may provide new, individual treatment options for patients with otherwise refractory tumors that do not respond anymore to standard chemotherapy., Competing Interests: Declaration of competing interest H.J.G. and T.E. are inventors of patent #EP18211231. The other authors do not have any potential conflict of interest in relation to this study., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Hundreds of nucleus-encoded mitochondrial precursor proteins are synthesized in the cytosol and imported into mitochondria in a post-translational manner. However, the early processes associated with mitochondrial protein targeting remain poorly understood. Here, we show that in Saccharomyces cerevisiae, the cytosol has the capacity to transiently store mitochondrial matrix-destined precursors in dedicated deposits that we termed MitoStores. Competitive inhibition of mitochondrial protein import via clogging of import sites greatly enhances the formation of MitoStores, but they also form during physiological cell growth on nonfermentable carbon sources. MitoStores are enriched for a specific subset of nucleus-encoded mitochondrial proteins, in particular those containing N-terminal mitochondrial targeting sequences. Our results suggest that MitoStore formation suppresses the toxic potential of aberrantly accumulating mitochondrial precursor proteins and is controlled by the heat shock proteins Hsp42 and Hsp104. Thus, the cytosolic protein quality control system plays an active role during the early stages of mitochondrial protein targeting through the coordinated and localized sequestration of mitochondrial precursor proteins., (© 2023 The Authors. Published under the terms of the CC BY 4.0 license.)

The DNMT3A DNA methyltransferase and MECP2 methylation reader are highly expressed in neurons. Both proteins interact via their DNMT3A-ADD and MECP2-TRD domains, and the MECP2 interaction regulates the activity and subnuclear localization of DNMT3A. Here, we mapped the interface of both domains using peptide SPOT array binding, protein pull-down, equilibrium peptide binding assays, and structural analyses. The region D529-D531 on the surface of the ADD domain was identified as interaction point with the TRD domain. This includes important residues of the histone H3 N-terminal tail binding site to the ADD domain, explaining why TRD and H3 binding to the ADD domain is competitive. On the TRD domain, residues 214-228 containing K219 and K223 were found to be essential for the ADD interaction. This part represents a folded patch within the otherwise largely disordered TRD domain. A crystal structure analysis of ADD revealed that the identified H3/TDR lysine binding pocket is occupied by an arginine residue from a crystallographic neighbor in the ADD apoprotein structure. Finally, we show that mutations in the interface of ADD and TRD domains disrupt the cellular interaction of both proteins in NIH3T3 cells. In summary, our data show that the H3 peptide binding cleft of the ADD domain also mediates the interaction with the MECP2-TRD domain suggesting that this binding site may have a broader role also in the interaction of DNMT3A with other proteins leading to complex regulation options by competitive and PTM specific binding., (© 2022 The Authors. Protein Science published by Wiley Periodicals LLC on behalf of The Protein Society.)

Current recommendations suggest neoadjuvant treatment in node-positive esophageal cancer or tumors staged T3 and upwards but some T2 N0 patients might benefit from neoadjuvant therapy. It is of clinical relevance to identify this subgroup. Loss of epithelial apicobasal polarity is a key factor in the development of invasive capabilities of carcinoma. The oncofetal stem/progenitor cell marker NOPE is expressed in adult depolarized murine hepatocytes and in murine/human hepatocellular carcinoma. We analyzed NOPE expression in 363 patients with esophageal adenocarcinoma using an RNA Scope Assay on a tissue microarray and correlated results with clinical data. Median follow-up was 57.7 months with a 5-year survival rate of 26.6%. NOPE was detectable in 32 patients (8.8%). In pT1/2 stages, NOPE expression was associated with a significantly reduced median OS of 6.3 months (95% CI 1.2-19.4 months), the median OS is not reached in the NOPE-negative group (calculated mean OS 117.1 months) (P = 0.012). In advanced tumor stages, a NOPE dependent survival difference was not detected. This is the first report of NOPE expression demonstrating a prognostic value in esophageal cancer. Early stage, NOPE positive patients are at a high risk of tumor progression and may benefit from neoadjuvant treatment analogous to advanced stage cancer., (© 2022. The Author(s).)

The formation of protein aggregates is a hallmark of neurodegenerative diseases. Observations on patient samples and model systems demonstrated links between aggregate formation and declining mitochondrial functionality, but causalities remain unclear. We used Saccharomyces cerevisiae to analyze how mitochondrial processes regulate the behavior of aggregation-prone polyQ protein derived from human huntingtin. Expression of Q97-GFP rapidly led to insoluble cytosolic aggregates and cell death. Although aggregation impaired mitochondrial respiration only slightly, it considerably interfered with the import of mitochondrial precursor proteins. Mutants in the import component Mia40 were hypersensitive to Q97-GFP, whereas Mia40 overexpression strongly suppressed the formation of toxic Q97-GFP aggregates both in yeast and in human cells. Based on these observations, we propose that the post-translational import of mitochondrial precursor proteins into mitochondria competes with aggregation-prone cytosolic proteins for chaperones and proteasome capacity. Mia40 regulates this competition as it has a rate-limiting role in mitochondrial protein import. Therefore, Mia40 is a dynamic regulator in mitochondrial biogenesis that can be exploited to stabilize cytosolic proteostasis., (© 2021 The Authors. Published under the terms of the CC BY 4.0 license.)

Most mitochondrial proteins are synthesized as precursors in the cytosol and post-translationally transported into mitochondria. The mitochondrial surface protein Tom70 acts at the interface of the cytosol and mitochondria. In vitro import experiments identified Tom70 as targeting receptor, particularly for hydrophobic carriers. Using in vivo methods and high-content screens, we revisit the question of Tom70 function and considerably expand the set of Tom70-dependent mitochondrial proteins. We demonstrate that the crucial activity of Tom70 is its ability to recruit cytosolic chaperones to the outer membrane. Indeed, tethering an unrelated chaperone-binding domain onto the mitochondrial surface complements most of the defects caused by Tom70 deletion. Tom70-mediated chaperone recruitment reduces the proteotoxicity of mitochondrial precursor proteins, particularly of hydrophobic inner membrane proteins. Thus, our work suggests that the predominant function of Tom70 is to tether cytosolic chaperones to the outer mitochondrial membrane, rather than to serve as a mitochondrion-specifying targeting receptor., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 The Author(s). Published by Elsevier Inc. All rights reserved.)

Most mitochondrial proteins are synthesized in the cytosol and subsequently translocated as unfolded polypeptides into mitochondria. Cytosolic chaperones maintain precursor proteins in an import-competent state. This post-translational import reaction is under surveillance of the cytosolic ubiquitin-proteasome system, which carries out several distinguishable activities. On the one hand, the proteasome degrades nonproductive protein precursors from the cytosol and nucleus, import intermediates that are stuck in mitochondrial translocases, and misfolded or damaged proteins from the outer membrane and the intermembrane space. These surveillance activities of the proteasome are essential for mitochondrial functionality, as well as cellular fitness and survival. On the other hand, the proteasome competes with mitochondria for nonimported cytosolic precursor proteins, which can compromise mitochondrial biogenesis. In order to balance the positive and negative effects of the cytosolic protein quality control system on mitochondria, mitochondrial import efficiency directly regulates the capacity of the proteasome via transcription factor Rpn4 in yeast and nuclear respiratory factor (Nrf) 1 and 2 in animal cells. In this review, we provide a thorough overview of how the proteasome regulates mitochondrial biogenesis., (© 2020 The Authors. FEBS Letters published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.)

The biogenesis of mitochondria requires the import of hundreds of precursor proteins. These proteins are transported post-translationally with the help of chaperones, meaning that the overproduction of mitochondrial proteins or the limited availability of chaperones can lead to the accumulation of cytosolic precursor proteins. This imposes a severe challenge to cytosolic proteostasis and triggers a specific transcription program called the mitoprotein-induced stress response, which activates the proteasome system. This coincides with the repression of mitochondrial proteins, including many proteins of the intermembrane space. In contrast, herein we report that the so-far-uncharacterized intermembrane space protein Mix23 is considerably up-regulated when mitochondrial import is perturbed. Mix23 is evolutionarily conserved and a homolog of the human protein CCDC58. We found that, like the subunits of the proteasome, Mix23 is under control of the transcription factor Rpn4. It is imported into mitochondria by the mitochondrial disulfide relay. Mix23 is critical for the efficient import of proteins into the mitochondrial matrix, particularly if the function of the translocase of the inner membrane 23 is compromised such as in temperature-sensitive mutants of Tim17. Our observations identify Mix23 as a novel regulator or stabilizer of the mitochondrial protein import machinery that is specifically up-regulated upon mitoprotein-induced stress conditions., Competing Interests: Conflict of interest—The authors declare that they have no conflicts of interest with the contents of this article., (© 2020 Zöller et al.)

In the version of this article originally published, parts of Figure 5 were misaligned because of a shift during production. In a, one data point was outside of the graph border. In b, axes lines were not connected, and graph lines did not reach the data points. In c and d, the axes lines were not connected. In e and g, the axes lines were not connected, and error bars and columns were not aligned. Shown below are the original and corrected versions of Figure 5. The errors have been corrected in the PDF and HTML versions of the paper.

The cytosolic accumulation of mitochondrial precursors is hazardous to cellular fitness and is associated with a number of diseases. However, it is not observed under physiological conditions. Individual mechanisms that allow cells to avoid cytosolic accumulation of mitochondrial precursors have recently been discovered, but their interplay and regulation remain elusive. Here, we show that cells rapidly launch a global transcriptional programme to restore cellular proteostasis after induction of a 'clogger' protein that reduces the number of available mitochondrial import sites. Cells upregulate the protein folding and proteolytic systems in the cytosol and downregulate both the cytosolic translation machinery and many mitochondrial metabolic enzymes, presumably to relieve the workload of the overstrained mitochondrial import system. We show that this transcriptional remodelling is a combination of a 'wideband' core response regulated by the transcription factors Hsf1 and Rpn4 and a unique mitoprotein-induced downregulation of the oxidative phosphorylation components, mediated by an inactivation of the HAP complex.

Background: Multimorbid older adults suffering from a long-term health condition like depression, diabetes mellitus type 2, dementia or frailty are at high risk of losing their autonomy. Disability and multimorbidity in the older population are associated with social inequality and lead to soaring costs. Our local, collaborative, stepped and personalised care management for older people with chronic diseases (LoChro-Care) aims at improving outcomes for older multimorbid patients with chronic conditions whose social and medical care must be improved., Methods: The study will evaluate the effects of LoChro-Care on functional health, depressive symptoms and satisfaction with care, resource utilisation as well as health costs in older persons with long-term conditions. The trial will compare the effectiveness of LoChro-Care and usual care in a cross-sectoral setting from hospital to community care. We will recruit 606 older adults (65+) admitted to local hospital inpatient or outpatient departments who are at risk of loss of independence. Half of them will be randomised to receive the LoChro-Care intervention, comprising seven to 16 contacts with chronic care managers (CCM) within 12 months. The hypothesis that LoChro-Care will result in better patient-centred outcomes will be tested through mixed-method process and outcome evaluation and valid measures completed at baseline and at 12 and 18 months. Cost-effectiveness analyses from the healthcare perspective will include incremental cost-effectiveness ratios., Discussion: The trial will provide evidence about the effectiveness of local, collaborative, stepped and personalised care management for multimorbid patients with more than one functional impairment or chronic condition. Positive results will be a first step towards the implementation of a systematic cross-sectoral chronic care management to facilitate the appropriate use of available medical and nursing services and to enhance self-management of older people., Trial Registration: German Clinical Trials Register (DRKS): DRKS00013904 ; Trial registration date: 02. February 2018.

Background: Internet- and mobile-based interventions are effective for the treatment of chronic pain. However, little is known about patients' willingness to engage with these types of interventions and how the uptake of such interventions can be improved., Objective: The aim of this study was to identify people's acceptance, uptake, and adherence (primary outcomes) with regard to an internet- and mobile-based intervention for chronic pain and the influence of an information video as an acceptance-facilitating intervention (AFI)., Methods: In this randomized controlled trial with a parallel design, we invited 489 individuals with chronic pain to participate in a Web-based survey assessing the acceptance of internet- and mobile-based interventions with the offer to receive an unguided internet- and mobile-based intervention for chronic pain after completion. Two versions of the Web-based survey (with and without AFI) were randomly sent to two groups: one with AFI (n=245) and one without AFI (n=244). Participants who completed the Web-based survey with or without AFI entered the intervention group or the control group, respectively. In the survey, the individuals' acceptance of pain interventions, measured with a 4-item scale (sum score ranging from 4 to 20), predictors of acceptance, sociodemographic and pain-related variables, and physical and emotional functioning were assessed. Uptake rates (log in to the intervention) and adherence (number of completed modules) to the intervention was assessed 4 months after intervention access. To examine which factors influence acceptance, uptake rate, and adherence in the internet- and mobile-based interventions, we conducted additional exploratory subgroup analyses., Results: In total, 57 (intervention group) and 58 (control group) participants in each group completed the survey and were included in the analyses. The groups did not differ with regard to acceptance, uptake rate, or adherence (P=.64, P=.56, P=.75, respectively). Most participants reported moderate (68/115, 59.1%) to high (36/115, 31.3%) acceptance, with 9.6% (11/115) showing low acceptance (intervention group: mean 13.91, SD 3.47; control group: mean 13.61, SD 3.50). Further, 67% (38/57, intervention group) and 62% (36/58, control group) had logged into the intervention. In both groups, an average of 1.04 (SD 1.51) and 1.14 (SD 1.90) modules were completed, respectively., Conclusions: The informational video was not effective with regard to acceptance, uptake rate, or adherence. Despite the high acceptance, the uptake rate was only moderate and adherence was remarkably low. This study shows that acceptance can be much higher in a sample participating in an internet- and mobile-based intervention efficacy trial than in the target population in routine health care settings. Thus, future research should focus not only on acceptance and uptake facilitating interventions but also on ways to influence adherence. Further research should be conducted within routine health care settings with more representative samples of the target population., Trial Registration: German Clinical Trial Registration DRKS00006183; http://www.drks.de/drks&#95;web/navigate.do ?navigationId=trial.HTML&TRIAL&#95;ID=DRKS00006183 (Archived by WebCite at http://www.webcitation.org/70ebHDhne)., (©Jiaxi Lin, Bianca Faust, David Daniel Ebert, Lena Krämer, Harald Baumeister. Originally published in the Journal of Medical Internet Research (http://www.jmir.org), 21.08.2018.)

Following publication of the original article [1], the author reported the errors in the figure and in text reference in the published article. These errors are detailed below. The original article has been corrected.

Within a global physical theory, a notion of locality allows us to find and justify information-processing primitives, like non-signalling between distant agents. Here, we propose exploring the opposite direction: to take agents as the basic building blocks through which we test a physical theory, and recover operational notions of locality from signalling conditions. First, we introduce an operational model for the effective state spaces of individual agents, as well as the range of their actions. We then formulate natural secrecy conditions between agents and identify the aspects of locality relevant for signalling. We discuss the possibility of taking commutation of transformations as a primitive of physical theories, as well as applications to quantum theory and generalized probability frameworks. This 'it from bit' approach establishes an operational connection between local actions and local observations, and gives a global interpretation to concepts like discarding a subsystem or composing local functions.This article is part of a discussion meeting issue 'Foundations of quantum mechanics and their impact on contemporary society'., (© 2018 The Authors.)

Background: Due to limited resources, waiting periods for psychotherapy are often long and burdening for those in need of treatment and the health care system. In order to bridge the gap between initial contact and the beginning of psychotherapy, web-based interventions can be applied. The implementation of a web-based depression intervention during waiting periods has the potential to reduce depressive symptoms and enhance well-being in depressive individuals waiting for psychotherapy., Methods: In a two-arm randomized controlled trial, effectiveness and acceptance of a guided web-based intervention for depressive individuals on a waitlist for psychotherapy are evaluated. Participants are recruited in several German outpatient clinics. All those contacting the outpatient clinics with the wish to enter psychotherapy receive study information and a depression screening. Those adults (age ≥ 18) with depressive symptoms above cut-off (CES-D scale > 22) and internet access are randomized to either intervention condition (treatment as usual and immediate access to the web-based intervention) or waiting control condition (treatment as usual and delayed access to the web-based intervention). At three points of assessment (baseline, post-treatment, 3-months-follow-up) depressive symptoms and secondary outcomes, such as quality of life, attitudes towards psychotherapy and web-based interventions and adverse events are assessed. Additionally, participants' acceptance of the web-based intervention is evaluated, using measures of intervention adherence and satisfaction., Discussion: This study investigates a relevant setting for the implementation of web-based interventions, potentially improving the provision of psychological health care. The results of this study contribute to the evaluation of innovative and resource-preserving health care models for outpatient psychological treatment., Trial Registration: This trial has been registered on 13 February 2017 in the German clinical trials register (DRKS); registration number DRKS00010282 .

Currently, changes in metabolic fluxes following consumption of stable isotope-enriched foods are usually limited to the analysis of postprandial kinetics of glucose. Kinetic information on a larger diversity of metabolites is often lacking, mainly due to the marginal percentage of fully isotopically enriched plant material in the administered food product, and hence, an even weaker 13 C enrichment in downstream plasma metabolites. Therefore, we developed an analytical workflow to determine weak 13 C enrichments of diverse plasma metabolites with conventional gas chromatography-mass spectrometry (GC-MS). The limit of quantification was increased by optimizing (1) the metabolite extraction from plasma, (2) the GC-MS measurement, and (3) most importantly, the computational data processing. We applied our workflow to study the catabolic dynamics of 13 C-enriched wheat bread in three human subjects. For that purpose, we collected time-resolved human plasma samples at 16 timepoints after the consumption of 13 C-labeled bread and quantified 13 C enrichment of 12 metabolites (glucose, lactate, alanine, glycine, serine, citrate, glutamate, glutamine, valine, isoleucine, tyrosine, and threonine). Based on isotopomer specific analysis, we were able to distinguish catabolic profiles of starch and protein hydrolysis. More generally, our study highlights that conventional GC-MS equipment is sufficient to detect isotope traces below 1% if an appropriate data processing is integrated., Competing Interests: D.M.J. is an employee of Unilever, which manufactures and markets consumer food products, including the flour used for the flatbreads in the human intervention study.

To date, it is well-established that mitochondrial dysfunction does not only play a vital role in cancer but also in other pathological conditions such as neurodegenerative diseases and inflammation. An important tool for the analysis of cellular metabolism is the application of stable isotope labeled substrates, which allow for the tracing of atoms throughout metabolic networks. While such analyses yield very detailed information about intracellular fluxes, the determination of compartment specific fluxes is far more challenging. Most approaches for the deconvolution of compartmented metabolism use computational models whereas experimental methods are rare. Here, we developed an experimental setup based on selective permeabilization of the cytosolic membrane that allows for the administration of stable isotope labeled substrates directly to mitochondria. We demonstrate how this approach can be used to infer metabolic changes in mitochondria induced by either chemical or genetic perturbations and give an outlook on its potential applications., (Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.)

One way to adjust the properties of materials is by changing its microstructure. This concept is not easily applicable on bulk metallic glasses (BMGs), because they do not consist of grains or different phases and so their microstructure is very homogeneous. One obvious way to integrate inhomogeneities is to produce bulk metallic glass composites (BMGCs). Here we show how to generate BMGCs via high-pressure torsion (HPT) starting from powders (amorphous Zr-MG and crystalline Cu). Using this approach, the composition can be varied and by changing the applied shear strains, the refinement of the microstructure is adjustable. This process permits to produce amorphous/crystalline composites where the scale of the phases can be varied from the micro- to the nanometer regime. Even mixing of the two phases and the generation of new metallic glasses can be achieved. The refinement of microstructure increases the hardness and a hardness higher than the initial BMG can be obtained.

The field of nutrition has evolved rapidly over the past century. Nutrition scientists and policy makers in the developed world have shifted the focus of their efforts from dealing with diseases of overt nutrient deficiency to a new paradigm aimed at coping with conditions of excess-calories, sedentary lifestyles and stress. Advances in nutrition science, technology and manufacturing have largely eradicated nutrient deficiency diseases, while simultaneously facing the growing challenges of obesity, non-communicable diseases and aging. Nutrition research has gone through a necessary evolution, starting with a reductionist approach, driven by an ambition to understand the mechanisms responsible for the effects of individual nutrients at the cellular and molecular levels. This approach has appropriately expanded in recent years to become more holistic with the aim of understanding the role of nutrition in the broader context of dietary patterns. Ultimately, this approach will culminate in a full understanding of the dietary landscape-a web of interactions between nutritional, dietary, social, behavioral and environmental factors-and how it impacts health maintenance and promotion.

Bone fracture with subsequent immobilization of the injured limb can cause complex regional pain syndrome (CRPS) in humans. Mechanisms of CRPS are still not completely understood but bone fracture with casting in mice leads to a similar post-traumatic inflammation as seen in humans and might therefore be an analog to human CRPS. In this article we report behavioral and spinal electrophysiological changes in mice that developed swelling of the paw, warming of the skin, and pain in the injured limb after bone fracture. The receptive field sizes of spinal neurons representing areas of the hind paws increased after trauma and recovered over time-as did the behavioral signs of inflammation and pain. Interestingly, both sides-the ipsi- and the contralateral limb-showed changes in mechanical sensitivity and neuronal network organization after the trauma. The characteristics of evoked neuronal responses recorded in the dorsal horn of the mice were similar between uninjured controls and fractured animals. However, we saw a caudal extension of the represented area of the hind paw in the spinal cord at the injured side and an occurrence of large receptive fields of wide dynamic range neurons. The findings in mice compare with human symptoms in CRPS with ipsi- and also contralateral allodynia and pain. In all mice tested, all signs subsided 12 weeks after trauma. Our data suggest a significant reorganization of spinal circuitry after limb trauma, in a degree more comprehensive than most models of neuropathies. This process seems to be reversible in the rodent., Perspective: The discovery of enlarged spinal neuronal receptive fields and caudal extension of the representation area of the injured body part, which subsides several weeks after a bone trauma in mice, might give hope to patients of CRPS if-in the future-we are able to translate the rodent recovery mechanisms to post-traumatic humans., (Copyright © 2017 American Pain Society. Published by Elsevier Inc. All rights reserved.)

Background: Common variants in the gene GATA binding protein 4 (GATA4) show association with alcohol dependence (AD). The aim of this study was to identify rare variants in GATA4 in order to elucidate the role of this gene in AD susceptibility. Identification of rare variants may provide a more complete picture of the allelic architecture at this risk locus., Methods: Sanger sequencing of all 6 coding exons of GATA4 was performed in 528 patients and 517 controls. Four in silico prediction tools were used to determine the effect of a DNA variant on the amino acid sequence and protein function. Five variants were included in the replication step. Of these, 4 were successfully genotyped in our replication cohort of 655 patients and 1,501 controls. All patients fulfilled DSM-IV criteria for AD, and all individuals were of German descent., Results: In the discovery step, 19 different heterozygous variants were identified. Four patient-specific and potentially functionally relevant variants were followed up. Only the variant S379S (c.1137C>T) remained patient specific (1/1,166 patients vs. 0/1,997 controls). None of the variants showed a statistically significant association with AD., Conclusions: The present study elucidated the role of GATA4 in AD susceptibility by identifying rare variants via Sanger sequencing and subsequent replication. Although novel patient-specific rare variants of GATA4 were identified, none received support in the independent replication step. However, given previous robust findings of association with common variants, GATA4 remains a promising candidate gene for AD., (Copyright © 2016 The Authors. Alcoholism: Clinical and Experimental Research published by Wiley Periodicals, Inc. on behalf of Research Society on Alcoholism.)

The oncogene DJ-1 has been originally identified as a suppressor of PTEN. Further on, loss-of-function mutations have been described as a causative factor in Parkinson's disease (PD). DJ-1 has an important function in cellular antioxidant responses, but its role in central metabolism of neurons is still elusive. We applied stable isotope assisted metabolic profiling to investigate the effect of a functional loss of DJ-1 and show that DJ-1 deficient neuronal cells exhibit decreased glutamine influx and reduced serine biosynthesis. By providing precursors for GSH synthesis, these two metabolic pathways are important contributors to cellular antioxidant response. Down-regulation of these pathways, as a result of loss of DJ-1 leads to an impaired antioxidant response. Furthermore, DJ-1 deficient mouse microglia showed a weak but constitutive pro-inflammatory activation. The combined effects of altered central metabolism and constitutive activation of glia cells raise the susceptibility of dopaminergic neurons towards degeneration in patients harboring mutated DJ-1. Our work reveals metabolic alterations leading to increased cellular instability and identifies potential new intervention points that can further be studied in the light of novel translational medicine approaches., (Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.)

Upon stimulation with Th1 cytokines or bacterial lipopolysaccharides, resting macrophages shift their phenotype toward a pro-inflammatory state as part of the innate immune response. LPS-activated macrophages undergo profound metabolic changes to adapt to these new physiological requirements. One key step to mediate this metabolic adaptation is the stabilization of HIF1α, which leads to increased glycolysis and lactate release, as well as decreased oxygen consumption. HIF1 abundance can result in the induction of the gene encoding pyruvate dehydrogenase kinase 1 (PDK1), which inhibits pyruvate dehydrogenase (PDH) via phosphorylation. Therefore, it has been speculated that pyruvate oxidation through PDH is decreased in pro-inflammatory macrophages. However, to answer this open question, an in-depth analysis of this metabolic branching point was so far lacking. In this work, we applied stable isotope-assisted metabolomics techniques and demonstrate that pyruvate oxidation is maintained in mature pro-inflammatory macrophages. Glucose-derived pyruvate is oxidized via PDH to generate citrate in the mitochondria. Citrate is used for the synthesis of the antimicrobial metabolite itaconate and for lipogenesis. An increased demand for these metabolites decreases citrate oxidation through the tricarboxylic acid cycle, whereas increased glutamine uptake serves to replenish the TCA cycle. Furthermore, we found that the PDH flux is maintained by unchanged PDK1 abundance, despite the presence of HIF1. By pharmacological intervention, we demonstrate that the PDH flux is an important node for M(LPS) macrophage activation. Therefore, PDH represents a metabolic intervention point that might become a research target for translational medicine to treat chronic inflammatory diseases., (© 2016 by The American Society for Biochemistry and Molecular Biology, Inc.)

Predictive in vitro models alternative to in vivo animal will have a significant impact in toxicology. Conventional 2D models do not reflect the complexity of a 3D organ resulting in discrepancies between experimental in vitro and in vivo data. Using 3D HepaRG organotypic cultures we tested four drugs (aflatoxin B1, amiodarone, valproic acid and chlorpromazine) for toxic effects and compared the results with 2D HepaRG and HepG2 cultures. We show that 3D HepaRG cultures are more sensitive than the other tested cultures to aflatoxin B1 which is only toxic upon metabolic activation in the liver. We observed that CYP3A4 activity is higher in the 3D HepaRG cultures compared to the 2D HepaRG cultures. Furthermore, we investigated repeated dose toxicity of chlorpromazine and assessed its effects on glucose and lactate metabolism. Sub-toxic concentrations of chlorpromazine induced significant metabolic changes in both 2D and 3D HepaRG cultures upon acute and repeated dose (3 doses) exposure. In summary, our data support the hypothesis that 3D cell culture models better mimic the in vivo tissue and improve cellular functionality. The 3D HepaRG organotypic cultures represent a high throughput system for drug toxicity screening. This system is therefore a promising tool in preclinical testing of human relevance which can allow reducing and/or replacing animal testing for drug adverse effects., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Membrane fusion at late endosomes and vacuoles depends on a conserved machinery, which includes Rab GTPases, their binding to tethering complexes and SNAREs. Fusion is initiated by the interaction of Rabs with tethering complexes. At the endosome, the CORVET complex interacts with the Rab5 GTPase Vps21, whereas the homologous HOPS complex binds the Rab7-like Ypt7 at the late endosome and vacuole. Activation of Ypt7 requires the recruitment of the Mon1-Ccz1 complex to the late endosome, which occurs via the CORVET complex. The interaction of Rab and the tethering complex is followed by the assembly of SNAREs, which leads to bilayer mixing. In this review, we will summarize our current knowledge on the mechanisms and regulation of endosome and vacuole membrane dynamics, and their role in organelle physiology., (Copyright © 2011 Elsevier GmbH. All rights reserved.)

Membrane fusion within the endomembrane system follows a defined order of events: membrane tethering, mediated by Rabs and tethers, assembly of soluble N-ethylmaleimide-sensitive factor (NSF) attachment protein receptor (SNARE) complexes, and lipid bilayer mixing. Here we present evidence that the vacuolar HOPS tethering complex controls fusion through specific interactions with the vacuolar SNARE complex (consisting of Vam3, Vam7, Vti1, and Nyv1) and the N-terminal domains of Vam7 and Vam3. We show that homotypic fusion and protein sorting (HOPS) binds Vam7 via its subunits Vps16 and Vps18. In addition, we observed that Vps16, Vps18, and the Sec1/Munc18 protein Vps33, which is also part of the HOPS complex, bind to the Q-SNARE complex. In agreement with this observation, HOPS-stimulated fusion was inhibited if HOPS was preincubated with the minimal Q-SNARE complex. Importantly, artificial targeting of Vam7 without its PX domain to membranes rescued vacuole morphology in vivo, but resulted in a cytokinesis defect if the N-terminal domain of Vam3 was also removed. Our data thus support a model of HOPS-controlled membrane fusion by recognizing different elements of the SNARE complex.

The two-domain, 325 residue outer membrane protein OmpA is one of the most abundant proteins of Escherichia coli, playing a role in the maintenance of the integrity of the cell surface. The N-terminal domain, consisting of about 170 amino acid residues, is embedded in the membrane, presumably in the form of a beta-barrel consisting of eight amphipathic transmembrane strands. Pairs of these proposed transmembrane strands were permuted at the DNA level, in order to dissect the process of membrane assembly. All three possible circular permutations led to variants, which were, in comparison with the wild-type protein, less efficiently assembled. In contrast, no membrane assembly could be detected in any of 18 non-circularly permuted variants. We take this as an indication that the "right" (wild-type) order of beta-strands is a necessary and sufficient prerequisite for at least partially successful membrane assembly. This may be the consequence of packing constraints and/or a failure to adopt the wild-type arrangement of beta-strands, which require crossing of the periplasmic turns.

E. coli phages of the T4 family (T4, TuIa, TuIb) recognize their cellular receptors with a C-terminal region of protein 37. This protein, common to all three phages, is present as a dimer located at the distal part of the long tail fibers and possesses a C-terminal domain consisting of 40 to 70 highly conserved C-terminal residues, followed by a variable region of 50 to 80 residues which is again followed by a highly conserved area. Protein 38, not being a component of the mature virion, is required for dimerization of protein 37; this represents a non-covalent association of a structural protein. Seven host range mutants of TuIa or TuIb were analyzed which were able to use proteinaceous receptors other than those recognized by their parents. All had suffered amino acid substitutions within the variable region. It is concluded that in all probability it is this region which interacts directly with the cellular receptors. Conditional mutants of T4 are known which, when propagated at the non-permissive temperature (42 degrees C), yield phage of normal morphology but these are more or less unable to adsorb to cells. The causative amino acid substitutions were found both downstream and upstream from the variable area. Distortion of it in the mutants could suggest a "snap-back" conformation of the tail fiber; the conserved C-terminal region may fold back and expose the variable region as a loop at the tip of the fiber. One of the phage mutants (L93), when grown at the permissive temperature, had lost the ability to use the OmpC porin (a receptor for T4) as a receptor. A secondary mutant, able to do so, was isolated. An additional mutation, leading to one amino acid substitution, had occurred in gene 38. This mutant gene acted in trans and caused a much enhanced temperature-sensitivity of infectivity without conferring temperature-sensitivity per se, i.e. the mutant protein 38 apparently altered the conformation of the receptor-recognizing area of the dimer of protein 37. A gene from phage lambda, about 40% identical to gene 38 of T4, complements gene 38 amber mutants. The corresponding protein also restored the ability of L93 to recognize OmpC but did not cause any such temperature-sensitivity. Hence, protein 38, classifying as a chaperone, appears to act instructively in conveying steric information to the target polypeptide.