610 Medizin und Gesundheit
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Human feline leukaemia virus subgroup C receptor-related proteins 1 and 2 (FLVCR1 and 2) are major facilitator superfamily transporters from the solute carrier family 49. Dysregulation of these ubiquitous transporters has been linked to various haematological and neurological disorders. While both FLVCRs were initially proposed to hold a physiological function in heme transport, subsequent studies questioned this notion. Here, we used structural, computational and biochemical methods and conclude that these two FLVCRs function as human choline transporters. We present cryo-electron microscopy structures of FLVCRs in different inward- and outward-facing conformations, captured in the apo state or in complex with choline in their translocation pathways. Our findings provide insights into the molecular framework of choline coordination and transport, largely mediated by conserved cation-π interactions, and further illuminate the conformational dynamics of the transport cycle. Moreover, we identified a heme binding site on the protein surface of the FLVCR2 N-domain, and observed that heme actively drives the conformational transitions of the protein. This auxiliary binding site might indicate a potential regulatory role of heme in the FLVCR2 transport mechanisms. Our work resolves the contested substrate specificity of the FLVCRs, and sheds light on the process of maintaining cellular choline homeostasis at the molecular level.
Der Einfluss von Trauma-aktiviertem platelet-rich fibrin auf mesenchymale Stammzellen in vitro
(2023)
Ziel dieser Arbeit war es, die Wechselwirkung zwischen MSC und PRF von Traumapatienten herauszuarbeiten und zu überprüfen, ob ein therapeutischer Einsatz der Kombination von MSC und Trauma-aktiviertem PRF in Frakturen sinnvoll erscheint.
Hierfür wurden MSC mit PRF über einen Zeitraum von 24 oder 120 Stunden co-inkubiert und die metabolische Aktivität, definierte Faktoren im Überstand (IL-6, CXCL10, VEGF und IDO-2) und die Genexpression ausgewählter Faktoren (MAPK8, MAPK14, IL-6, CXCL10,IDO-1, TNFAIP6, VEGFA, RUNX2 und COL1A) in bis zu drei Messzeitpunkten überprüft. Die Versuche erfolgten vergleichend mit Traumapatienten und gesunden Probanden.
Es konnte gezeigt werden, dass Trauma-aktiviertes PRF verglichen mit PRF von gesunden Probanden die metabolische Aktivität von MSC nach 120 Stunden erhöhen kann. Zudem stellten sich erhöhte Werte des inflammatorischen Faktors IL-6 im Überstand bei Traumapatienten nach 24 und 72 Stunden dar, welche bis zum dritten Messzeitpunkt nach 120 Stunden wieder abfielen. Ein ähnlicher Verlauf zeigte sich bei den Messwerten von VEGFA, einem Faktor, welcher eine große Rolle bei der Angiogenese spielt.
Über alle Messzeitpunkte hinweg konnten niedrigere Werte in der Genexpression von COL1A und RUNX2 im Vergleich zu den Kontrollmessungen erfasst werden.
Die Ergebnisse zeigen, dass im Vergleich zu PRF gesunder Probanden Trauma-aktiviertes PRF die Möglichkeit besitzt, die Proliferation von MSC zu stimulieren. Außerdem wurde beobachtet, dass Trauma-aktiviertes PRF ein inflammatorisches und angiogenetisches Potenzial nach 72 Stunden besitzt, welches nach 120 Stunden allerdings wieder abfällt. Möglicherweise kann durch den Einfluss von MSC der Inflammation entgegengewirkt werden. Ein signifikantes osteogenes Potenzial des Trauma-aktivierten PRFs konnte zu keinem Messzeitpunkt nachgewiesen werden.
Inwiefern der Einsatz von MSC in Kombination mit Trauma-aktiviertem PRF die Frakturheilung verbessern kann, konnte in der vorliegenden Studie nicht eindeutig geklärt werden. Es kann allerdings davon ausgegangen werden, dass die Implantation von Trauma-aktiviertem PRF und MSC in den Defekt erst nach Verstreichen einer bestimmten Zeitperiode nach dem Trauma durchgeführt werden sollte. Die Gründe für diese Annahme sind das erhöhte proliferative und verringerte inflammatorische Potenzial im Verlauf. Eine Ursache für das Fehlen der osteogenen Differenzierung der MSC kann der kurze Messzeitraum sein.
Weitere Studien sollten folgen, um das Potenzial der Kombination von MSC und Trauma-aktiviertem PRF als therapeutisches Verfahren zu überprüfen.
A Corrigendum on "SARS-CoV-2 Omicron variants: burden of disease, impact on vaccine effectiveness and need for variant-adapted vaccines" by Pather S, Madhi SA, Cowling BJ, Moss P, Kamil JP, Ciesek S, Muik A and Türeci Ö (2023). . 14:1130539. doi: 10.3389/fimmu.2023.1130539
The ongoing COVID-19 pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is partly under control by vaccination. However, highly potent and safe antiviral drugs for SARS-CoV-2 are still needed to avoid development of severe COVID-19. We report the discovery of a small molecule, Z-Tyr-Ala-CHN2, which was identified in a cell-based antiviral screen. The molecule exerts sub-micromolar antiviral activity against SARS-CoV-2, SARS-CoV-1, and human coronavirus 229E. Time-of-addition studies reveal that Z-Tyr-Ala-CHN2 acts at the early phase of the infection cycle, which is in line with the observation that the molecule inhibits cathepsin L. This results in antiviral activity against SARS-CoV-2 in VeroE6, A549-hACE2, and HeLa-hACE2 cells, but not in Caco-2 cells or primary human nasal epithelial cells since the latter two cell types also permit entry via transmembrane protease serine subtype 2 (TMPRSS2). Given their cell-specific activity, cathepsin L inhibitors still need to prove their value in the clinic; nevertheless, the activity profile of Z-Tyr-Ala-CHN2 makes it an interesting tool compound for studying the biology of coronavirus entry and replication.
Trauma ist in Deutschland und weltweit eine der häufigsten Todesursache bei Personen unter 55 Jahren. Eine traumatische Verletzung von Gewebe führt zur Freisetzung von sogenannten damage-associated molecular patterns (DAMPs), die eine Entzündungskaskade auslösen, welche die Organfunktion negativ beeinträchtigt. Dies führt bei anhaltender Entzündung zunächst zu Organdysfunktion, was im weiteren Verlauf zu Organversagen führt und im Spätstadium im multiplen Organversagen (MOF) enden kann. In den meisten Fällen ist die inflammatorische Antwort des Immunsystems auf das Trauma adäquat und entspricht einem gut koordinierten Netzwerk von Immunzellen, Zytokinen und Chemokinen, welches zur Wiederherstellung des geschädigten Gewebes führt. Wenn dieses Netzwerk jedoch nicht im Gleichgewicht ist, kann die Entzündungsreaktion durch eine sogenannte feed-forward-loop von Inflammation und Gewebeschäden verstärkt werden. Wenn dieser Prozess systemisch wird, spricht man vom systemic inflammatory response syndrome (SIRS). Auf der anderen Seite gibt es ein Gegenstück zu SIRS, nämlich das sogenannte compensatory anti-inflammatory response syndrome (CARS). Auch ein Überschießen von CARS kann den Verlauf der posttraumatischen Inflammation negativ beeinträchtigen. Sowohl eine Verschiebung in Richtung von SIRS als auch in Richtung von CARS kann zu Organfunktionsstörungen, nosokomialen Infektionen und letztendlich zum Tod führen. Daher ist eine ausgeglichene, frühe posttraumatische Immunantwort für ein gutes Outcome von entscheidender Bedeutung.
Monozyten nehmen eine kritische Stellung sowohl in der primären Immunantwort nach Trauma als auch nach Infektion ein. Durch die Oberflächenexpression von diversen sogenannten pattern-recocnition receptors (PRRs), insbesondere Toll-Like-Rezeptoren (TLRs), können Monozyten Pathogene und sogenannte pathogen-associated molecular patterns (PAMPs) erkennen und neutralisieren. Darüber hinaus können Monozyten PAMPs über major histocompatibility complex class II -Moleküle (MHC-II) dem adaptiven Immunsystem präsentieren und somit als zelluläre Verbindung zwischen dem angeborenen und dem adaptiven Immunsystem fungieren. TLR2, eine Untergruppe der TLRs, ist einer der Hauptrezeptoren für PAMPs von grampositiven Bakterien wie S. aureus, dem Hauptkeim für posttraumatische Wundinfektionen und nosokomiale Infektionen.
Es gibt immer noch keinen Konsens über den Einfluss von Trauma auf die Funktion von Monozyten und deren Expressionsprofil von PRRs und MHC-II-Molekülen nach einem Trauma. Verschiedene Studien berichten von einer beeinträchtigten TLR2-Expression bei Monozyten nach Trauma, während sich in anderen Studien eine konstante oder sogar erhöhte TLR2-Expression bei Monozyten nach Trauma gezeigt hat. Die Daten sind auch in Bezug auf die posttraumatische Phagozytoseleistung von Monozyten unbeständig. Während in einigen Studien im frühen posttraumatischen Verlauf im Vergleich zu gesunden Probanden von einer erhöhten Anzahl zirkulierender Monozyten mit erhaltener Phagozytoseleistung berichtet wird, zeigten andere Studien eine verminderte Phagozytoseleistung von Monozyten nach Trauma. Darüber hinaus gibt es widersprüchliche Ergebnisse bezüglich der Fähigkeit von Monozyten, proinflammatorische Zytokine wie Interleukin (IL)-1β oder Tumor necrosis factor (TNF)-α freizusetzen.
Zusammengefasst ist die derzeitige Studienlage widersprüchlich. Zusätzlich wurde es in bisherigen Studien verpasst, die Kombination verschiedener Aspekte wie die Funktionalität der Monozyten in Bezug auf Phänotypisierung oder funktioneller Assays und deren Beobachtung über einen längeren Zeitraum miteinzubeziehen.
Die unterschiedlichen Ergebnisse der bisherigen Studien könnte durch die Natur des Polytraumas bedingt sein, beispielsweise durch verschiedene Verletzungsmuster und Verletzungsschwere, die unterschiedliche Erstversorgung am Unfallort, die Zeit bis zur Ankunft in der Notaufnahme oder die unterschiedliche primäre und definitive Versorgung der Verletzungen. All das kann zu einer unterschiedlichen Antwort des Immunsystems und somit zu unterschiedlichen Outcomes führen.
Daher war es Ziel unserer Studie, die oben genannten Variablen zu eliminieren, und eine kontrollierte Polytraumastudie am Schweinemodell durchzuführen, um die Funktionalität von Schweinemonozyten über einen Zeitraum von 72 Stunden nach Trauma zu untersuchen.
Zusätzlich und im Gegensatz zu früheren Studien haben wir die Phänotypisierung von Monozyten (TLR2 und MHC-II [SLA-DR]) mit einem funktionellen Phagozytose-Assay kombiniert und deren direkte Assoziation in einem unabhängigen Assay analysiert.
Peripheres Blut wurde vor (-1h) und direkt nach der Induktion des Polytraumas (PT) (0h) entnommen, sowie 3,5h, 5,5h, 24h und 72h später. Die Expression von H(S)LA-DR und TLR2 auf Schweine-Monozyten wurde untersucht. Außerdem wurde die Phagozytierungsaktivität von Schweinemonozyten gemessen.
Darüber hinaus wurden aus mechanistischen Gründen Blutproben von 10 gesunden Schweinen zunächst einem TLR2-neutralisierenden Antikörper und anschließend S. aureus-Partikeln ausgesetzt, bevor die Phagozytoseleistung der Monozyten untersucht wurde.
Die Anzahl der CD14 + -Monozyten aller zirkulierenden Leukozyten blieb während des Beobachtungszeitraums konstant, während der Prozentsatz der CD14 + H(S)LA-DR + -Monozyten direkt, 3,5h und 5,5h nach dem Trauma signifikant abnahm. Der Prozentsatz von TLR2 + exprimierenden Zellen aus allen Monozyten verringerte sich direkt, 3,5h und 5,5h nach dem Trauma signifikant. Der Prozentsatz der phagozytierenden Monozyten nahm direkt nach Trauma ab und blieb in den ersten 3,5 Stunden nach dem Trauma niedriger, stieg jedoch nach 24 Stunden an. Die Antagonisierung von TLR2 verringerte signifikant die Phagozytoseleistung der Monozyten.
Sowohl der verringerte Prozentsatz der aktivierten als auch der TLR2-exprimierenden Monozyten blieb bestehen, solange die verringerte Phagozytoseleistung beobachtet wurde.
Darüber hinaus führte auch die Neutralisation von TLR2 zu einer verminderten Phagozytoseleistung. Daher nehmen wir an, dass eine verringerte TLR2-Expression für die verringerte Phagozytoseleistung verantwortlich ist.
Background: Developmentally adapted cognitive processing therapy (D-CPT) is an effective treatment for posttraumatic stress disorder (PTSD) in adolescents and young adults. It is unclear if therapeutic adherence and competence in D-CPT are associated with higher PTSD treatment gains.
Objective: To assess if higher therapeutic adherence and competence in D-CPT are associated with higher symptom reduction of PTSD in adolescents and young adults, while controlling for therapeutic alliance.
Participants and setting: Participants were 38 patients (aged 14–21 years; M = 17.61 years, SD = 2.42 years) of a multicenter randomized controlled trial in which the efficacy of D-CPT was compared to a waitlist with treatment advice.
Methods: Videotaped therapy sessions were rated using validated ratings scales to assess adherence and competence. Therapeutic alliance was assessed via weekly patient ratings. We used hierarchical linear modelling to assess the relationship of adherence and competence on PTSD symptoms being measured by both clinician and patient while controlling for alliance.
Results: Neither adherence nor competence were related to treatment outcomes in clinician or patient rated PTSD symptom severity. Higher alliance was associated with a lower symptom severity at 12 months posttreatment in both clinician and patient rated PTSD symptoms.
Conclusions: In this study of young adults with PTSD, who were treated with D-CPT by well-trained therapists, therapeutic adherence and competence were not related to treatment outcome. This might be explained by a lack of range in therapist adherence and competence. Therapeutic alliance had a positive effect on PTSD symptom severity.
The role of lncRNAs in the CVS and the endothelium is highly diverse and has been subject to a substantial amount of research over the last decade. The identification of lncRNAs as clinically relevant biomarkers and as co-regulatory molecules let to the appreciation of the functional relevance of lncRNAs.
In the present study, LINC00607 was identified as an endothelial-enriched, human-specific lncRNA. With its distinct functions, LINC00607 maintains and supports the endothelial homeostasis especially in response to VEGF-A signalling.
In the first part of this study, LINC00607 was functionally characterized in human endothelial cells. LINC00607 is highly and specifically expressed in endothelial cells and is differentially regulated in CVDs. Depletion of LINC00607 resulted in decreased angiogenic sprouting, reduced integration of ECs in a newly formed vascular network in vivo, enhanced endothelial migration and differential expression of many important genes for endothelial cell homeostasis. Functionally, LINC00607 maintains ERG-driven endothelial gene expression programs through BRG1. BRG1 secures stably accessible enhancer regions as well as TSS of ERG target genes, thus enabling transcription of endothelial gene programs.
The second part of this study proposes an additional mode of action for LINC00607. The strongly impaired response to VEGF-A after LINC00607 KO can only be partially explained by its’ expression control of ERG target genes. It rather appears that LINC00607 is involved in the control of alternative splicing of VEGF receptor FLT1. The differential splicing of FLT1 produces the anti-angiogenic soluble isoform of FLT1. Even though further validation is needed to uncover the underlying mechanism, there is the potential of a more general role of LINC00607 in splicing control through BRG1. As AS of FLT1 is a clinical marker in preeclampsia, LINC00607 might qualify to be an additional marker for the onset and manifestation of the pregnancy disorder.
Taken together, LINC00607 is a target in future for molecular therapy in CVD to restore a healthy endothelial phenotype and has the potential to serve as a biomarker in preeclampsia.
Behavioral and psychological syndromes such as depression and psychosis often occur along with cognitive (esp. executive) deficits in vascular cognitive disorder (VCD) in the elderly. We present the case of an 85-year-old woman with deficits in executive functions as well as a persistent and clearly circumscribed paranoid hallucinatory syndrome (most probably due to VCD) which could not be adequately treated with antipsychotic medication. The patient also suffered from severe depression (independent of psychotic symptoms). Both psychosis and depression were successfully managed in a home treatment based on Flexible Assertive Community Treatment (FACT). Interestingly, a thematic association between the delusional contents and early childhood traumata could be reconstructed, and late-onset trauma-related symptoms could be successfully treated with cognitive-behavioral therapy (CBT) as well. In sum, behavioral management of psychotic syndromes is possible in the absence of adequate pharmacological treatment options, and multiprofessional and person-centered home treatment may be successful in the elderly, even in severe and complex disorders.
Febrile neutropenia is a common infectious complication in children and adolescents receiving chemotherapy for cancer, requiring immediate hospitalisation and empirical antibacterial therapy. The risk for a severe infection increases with lower neutrophil counts, but other factors such as underlying malignancy, remission state or the genetic background might also impact on the risk and severity of infection. Initial antibacterial treatment as well as modification and cessation of therapy depends on clinical performance, microbiological findings and haematological recovery. Although paediatric specific guidelines have been developed in the last decade, a number of questions are still unsolved. This article gives an overview on diagnostics and management of paediatric patients presenting with febrile neutropenia, on research gaps and will speculate on future perspective.
In this comprehensive review, we will dissect the impact of research on proteoglycans focusing on recent developments involved in their synthesis, degradation, and interactions, while critically assessing their usefulness in various biological processes. The emerging roles of proteoglycans in global infections, specifically the SARS-CoV-2 pandemic, and their rising functions in regenerative medicine and biomaterial science have significantly affected our current view of proteoglycans and related compounds. The roles of proteoglycans in cancer biology and their potential use as a next-generation protein-based adjuvant therapy to combat cancer is also emerging as a constructive and potentially beneficial therapeutic strategy. We will discuss the role of proteoglycans in selected and emerging areas of proteoglycan science, such as neurodegenerative diseases, autophagy, angiogenesis, cancer, infections and their impact on mammalian diseases.
A 24-year-old patient from Cameroon presented to our hospital because of a foreign structure in her left eye. To our knowledge, for the first time, fluorescent microscopy revealed motile microfilariae, and the diagnosis of loiasis was established. Despite substantial microfilaremia, eosinophilia only unmasked after the initiation of antiparasitic therapy.
Highlights
• Open pulmonary tuberculosis patient discharge policy was not reviewed for decades.
• After smear-negativity conversion, substantial cultural positivity may remain.
• It remains unclear, whether smear-negative patients still may be infective.
• The clinical relevance of this finding warrants further investigation.
Abstract
Objectives: Patients with open pulmonary tuberculosis (opTB) are subject to strict isolation rules. Sputum smear microscopy is used to determine infectivity, but sensitivity is lower than for culture. This study aimed to investigate the clinical relevance of this mismatch in contemporary settings.
Methods: Differential results between microscopy and culture were determined at the time of microscopic sputum conversion, from all patients with opTB between 01/2013 and 12/2017. In addition, data on HIV, multi/extensive drug-resistant TB status, time to smear- and cultural-negativity conversion were analyzed; and a Kaplan-Meier curve was developed.
Results: Of 118 patients with opTB, 58 had demographic data available for microbiological and clinical follow-up analysis; among these, 26 (44.8%) had still at least one positive culture result. Median time from opTB-treatment initiation to full microscopic sputum- or culture conversion, was 16.5 days (range 2-105), and 20 days (1-105), respectively (median difference: +3.5 days). Sixteen days after de-isolation, >90% had converted culturally. HIV- or multi/extensive drug-resistant TB status did not impact conversion time.
Conclusion: When patients with opTB were de-isolated after 3 negative sputum smear microscopy tests, a substantial part still revealed cultural growth of Mycobacterium tuberculosis complex, but it remains unclear, whether smear-negative and culturally-positive individuals on therapy are really infective. Thus, the clinical relevance of this finding warrants further investigation.
Abnormal venous atrial (VA) connections present a congenital heart disease (CHD) challenge for pediatric cardiologists. Fully anatomical evaluation is very difficult in prenatal and perinatal follow-up, but it has a profound impact on surgical correction and outcome. The echocardiogram is first-line imaging and represents the gold standard tool for simple abnormal VA connection. CT and MRI are mandatory for more complex heart disease and “nightmare cases”. 3D post-processing of volumetric CT and MRI acquisition helps to clarify anatomical relationships and allows for the creation of 3D printing models that can become crucial in customizing surgical strategy.
Approximately 80 % of persistent wound infections are affected by the presence of bacterial biofilms, resulting in a severe clinical challenge associated with prolonged healing periods, increased morbidity, and high healthcare costs. Unfortunately, in vitro models for wound infection research almost exclusively focus on early infection stages with planktonic bacteria. In this study, we present a new approach to emulate biofilm-infected human wounds by three-dimensional human in vitro systems. For this purpose, a matured biofilm consisting of the clinical key wound pathogen Pseudomonas aeruginosa was pre-cultivated on electrospun scaffolds allowing for non-destructive transfer of the matured biofilm to human in vitro wound models. We infected tissue-engineered human in vitro skin models as well as ex vivo human skin explants with the biofilm and analyzed structural tissue characteristics, biofilm growth behavior, and biofilm-tissue interactions. The structural development of biofilms in close proximity to the tissue, resulting in high bacterial burden and in vivo-like morphology, confirmed a manifest wound infection on all tested wound models, validating their applicability for general investigations of biofilm growth and structure. The extent of bacterial colonization of the wound bed, as well as the subsequent changes in molecular composition of skin tissue, were inherently linked to the characteristics of the underlying wound models including their viability and origin. Notably, the immune response observed in viable ex vivo and in vitro models was consistent with previous in vivo reports. While ex vivo models offered greater complexity and closer similarity to the in vivo conditions, in vitro models consistently demonstrated higher reproducibility. As a consequence, when focusing on direct biofilm-skin interactions, the viability of the wound models as well as their advantages and limitations should be aligned to the particular research question of future studies. Altogether, the novel model allows for a systematic investigation of host-pathogen interactions of bacterial biofilms and human wound tissue, also paving the way for development and predictive testing of novel therapeutics to combat biofilm-infected wounds.
For medicine to fulfill its promise of personalized treatments based on a better understanding of disease biology, computational and statistical tools must exist to analyze the increasing amount of patient data that becomes available. A particular challenge is that several types of data are being measured to cope with the complexity of the underlying systems, enhance predictive modeling and enrich molecular understanding.
Here we review a number of recent approaches that specialize in the analysis of multimodal data in the context of predictive biomedicine. We focus on methods that combine different OMIC measurements with image or genome variation data. Our overview shows the diversity of methods that address analysis challenges and reveals new avenues for novel developments.
Mitochondrial matrix peptidase CLPP is crucial during cell stress. Its loss causes Perrault syndrome type 3 (PRLTS3) with infertility, neurodegeneration and growth deficit. Its target proteins are disaggregated by CLPX, which also regulates heme biosynthesis via unfolding ALAS enzyme, providing access of pyridoxal-5’-phosphate (PLP). Despite efforts in diverse organisms with multiple techniques, CLPXP substrates remain controversial. Here, avoiding recombinant overexpression, we employed complexomics in mitochondria from three mouse tissues to identify endogenous targets. CLPP absence caused accumulation and dispersion of CLPX-VWA8 as AAA+ unfoldases, and of PLPBP. Similar changes and CLPX-VWA8 comigration were evident for mitoribosomal central protuberance clusters, translation factors like GFM1-HARS2, RNA granule components LRPPRC-SLIRP, and enzymes OAT-ALDH18A1. Mitochondrially translated proteins in testis showed reductions to <30% for MTCO1-3, misassembly of complex-IV supercomplex, and accumulated metal-binding assembly factors COX15-SFXN4. Indeed, heavy metal levels were increased for iron, molybdenum, cobalt and manganese. RT-qPCR showed compensatory downregulation only for Clpx mRNA, most accumulated proteins appeared transcriptionally upregulated. Immunoblots validated VWA8, MRPL38, MRPL18, GFM1 and OAT accumulation. Coimmunoprecipitation confirmed CLPX binding to MRPL38, GFM1 and OAT, so excess CLPX and PLP may affect their activity. Our data elucidate mechanistically the mitochondrial translation fidelity deficits, which underlie progressive hearing impairment in PRLTS3.
Viruses that carry a positive-sense, single-stranded (+ssRNA) RNA translate their genomes soon after entering the host cell to produce viral proteins, with the exception of retroviruses. A distinguishing feature of retroviruses is reverse transcription, where the +ssRNA genome serves as a template to synthesize a double-stranded DNA copy that subsequently integrates into the host genome. As retroviral RNAs are produced by the host cell transcriptional machinery and are largely indistinguishable from cellular mRNAs, we investigated the potential of incoming retroviral genomes to directly express proteins. Here we show through multiple, complementary methods that retroviral genomes are translated after entry. Our findings challenge the notion that retroviruses require reverse transcription to produce viral proteins. Synthesis of retroviral proteins in the absence of productive infection has significant implications for basic retrovirology, immune responses and gene therapy applications.
Viruses that carry a positive-sense, single-stranded RNA translate their genomes after entering the host cell to produce viral proteins, with the exception of retroviruses. A distinguishing feature of retroviruses is reverse transcription, where the ssRNA genome serves as a template to synthesize a double-stranded DNA copy that subsequently integrates into the host genome. As retroviral RNAs are produced by the host transcriptional machinery and are largely indistinguishable from cellular mRNAs, we investigated the potential of incoming retroviral genomes to express proteins. Here we show through various biochemical methods that HIV-1 genomes are translated after entry, in case of minimal or full-length genomes, envelopes using different cellular entry pathways and in diverse cell types. Our findings challenge the dogma that retroviruses require reverse transcription to produce viral proteins. Synthesis of retroviral proteins in the absence of productive infection has significant implications for basic retrovirology, immune responses and gene therapy applications.
Human feline leukemia virus subgroup C receptor-related proteins 1 and 2 (FLVCR1 and 2) are members of the major facilitator superfamily1. Their dysfunction is linked to several clinical disorders, including PCARP, HSAN, and Fowler syndrome2–7. Earlier studies concluded that FLVCR1 may function as a putative heme exporter8–12, while FLVCR2 was suggested to act as a heme importer13, yet conclusive biochemical and detailed molecular evidence remained elusive for the function of both transporters14–17. Here, we show that FLVCR1 and FLVCR2 facilitate the transport of choline and ethanolamine across human plasma membranes, utilizing a concentration-driven substrate translocation process. Through structural and computational analyses, we have identified distinct conformational states of FLVCRs and unraveled the coordination chemistry underlying their substrate interactions. Within the binding pocket of both transporters, we identify fully conserved tryptophan and tyrosine residues holding a central role in the formation of cation-π interactions, essential for choline and ethanolamine selectivity. Our findings not only clarify the mechanisms of choline and ethanolamine transport by FLVCR1 and FLVCR2, enhancing our comprehension of disease-associated mutations that interfere with these vital processes, but also shed light on the conformational dynamics of these MFS-type proteins during the transport cycle.
Zielsetzung der vorliegenden Dissertation war die Validierung einer deutschsprachigen Version, der bereits für andere Sprachen validierten Repetitiven Verhaltensskala – Revidiert (RBS-R) für Kinder- und Jugendliche im Alter zwischen 4-17 Jahren. Die RBS-R ist ein aus 43 Items bestehendes, gut untersuchtes Screeninginstrument, welches restriktives, repetitives Verhalten (RRV) erfasst. RRV stellen einen heterogenen Symptomkomplex dar, welcher durch stereotype motorische, sprachliche und kognitive Verhaltensmuster gekennzeichnet ist.
RRV sind neben Abweichungen sozialer Interaktion und Kommunikation eines der Hauptkriterien für eine Autismus-Spektrum-Störung (ASS). RRV sind jedoch nicht nur Teil des Symptomspektrums einer ASS, sondern treten häufig auch bei anderen psychiatrischen Erkrankungen (PE), einer Intelligenzminderung (IM) oder bei altersentsprechend entwickelten Kindern (AE), beispielsweise in bestimmten Phasen des Heranwachsens auf. Bisher wurde die RBS-R fast ausschließlich an ASS untersucht. Es besteht die Notwendigkeit für ein valides und zuverlässiges Messinstrument für den deutschen Sprachraum, welches RRV hinsichtlich ihrer Heterogenität transdiagnostisch erfassen und den Schweregrad der Ausprägung ermitteln kann. Das Ziel dieser Arbeit war, dies durch eine möglichst große und heterogene Stichprobe (n= 948) mit Inklusion von Probanden mit einer ASS (n= 218), IM (n= 120), PE (n= 166) und AE (n = 444) zu gewährleisten. Zur Überprüfung der psychometrischen Eignung erfolgte nach der Adaptation der RBS-R für den deutschen Sprachraum eine Evaluation gemäß den Prinzipien der klassischen Testtheorie, insbesondere hinsichtlich Validität und Reliabilität. Darüber hinaus erfolgte unter Einbeziehung der vier Stichproben eine explorative Faktoranalyse (EFA) um eine geeignete Faktorlösung zu finden. Diese sollte sowohl den Anspruch Klinisch-Tätiger als auch Forschender erfüllen und eine Anwendung der RBS-R auch außerhalb von ASS ermöglichen. Zudem sollten RRV in den unterschiedlichen Kohorten, sowie in Bezug auf Alters- und Geschlechtseffekte bei ASS untersucht werden.
Die Bedeutung der Ergebnisse dieser Arbeit ist, ob der schon breit erforschten Eigenschaften der RBS-R, vor allem im Kontext mit den vorliegenden Studien zu betrachten. Einzigartig für diese deutschsprachige Validierung der RBS-R ist die Diversität und große Anzahl der teilnehmenden Probanden.
Die vorliegende Validierung der RBS-R erbrachte gute Ergebnisse für die Reliabilität und konvergente Validität der Skalen, vergleichbar mit den Ergebnissen vorheriger Studien. Die untersuchte Itemschwierigkeit war relativ gering, was durch die geringe Antwortvarianz in der großen Kohorte der AE erklärbar ist. Die Itemgesamtkorrelation zeigte gute Werte. Auch Items mit niedrigen Ergebnissen für die Itemzustimmung wurden nicht exkludiert. Dies geschah konkordant zum Vorgehen vorheriger Studien und hatte den Hintergrund, eine internationale Vergleichbarkeit des RBS-R beizubehalten. Die EFA bestätigte die gute Anwendbarkeit einer Vielzahl von Faktorlösungen. Wir legten eine 4-Faktorlösung für die deutschsprachige Version der RBS-R fest. Grundlage hierfür war das Bestreben die ursprünglichen Subskalen für eine internationale Vergleichbarkeit so getreu wie möglich beizubehalten. Die 4 Faktoren bildeten treffend RRV niedriger und hoher Ordnung ab. Gegen eine 5- oder 6-Faktorlösung sprach hierbei, dass dies zu Subskalen mit wenig Items und schwachen psychometrischen Eigenschaften führte (Lam et al, kritisiert durch Georgiades et al). Gegen eine 2- oder 3-Faktorlösung sprach die eingeschränkte klinische Interpretierbarkeit durch Subskalen mit vielen kumulierten Items. Hinsichtlich der Kohortenzugehörigkeit kann der RBS-R übereinstimmend mit Fulceri et al valide zwischen RRV in ASS und AE unterscheiden. Eine valide Unterscheidung zwischen ASS, PE und IM konnten wir nicht verifizieren. Betreffend der untersuchten Alterseffekts bestätigte sich, dass RRV bei ASS mit dem Alter abnehmen. Im Gegensatz zu einigen vorherigen Studien, welche keinen Geschlechtsunterschied bezüglich RRV bei ASS fanden, zeigten die männlichen Probanden der ASS Kohorte bei uns häufiger und ausgeprägter RRV.
Background: Spondylodiscitis is a potentially life-threatening infection of the intervertebral disk and adjacent vertebral bodies, with a mortality rate of 2–20%. Given the aging population, the increase in immunosuppression, and intravenous drug use in England, the incidence of spondylodiscitis is postulated to be increasing; however, the exact epidemiological trend in England remains unknown.
Objective: The Hospital Episode Statistics (HES) database contains details of all secondary care admissions across NHS hospitals in England. This study aimed to use HES data to characterise the annual activity and longitudinal change of spondylodiscitis in England.
Methods: The HES database was interrogated for all cases of spondylodiscitis between 2012 and 2019. Data for the length of stay, waiting time, age-stratified admissions, and ‘Finished Consultant Episodes’ (FCEs), which correspond to a patient's hospital care under a lead clinician, were analysed.
Results: In total, 43135 FCEs for spondylodiscitis were identified between 2012 and 2022, of which 97.1% were adults. Overall admissions for spondylodiscitis have risen from 3 per 100,000 population in 2012/13 to 4.4 per 100,000 population in 2020/21. Similarly, FCEs have increased from 5.8 to 10.3 per 100,000 population, in 2012–2013 and 2020/21 respectively. The highest increase in admissions from 2012 to 2021 was recorded for those aged 70–74 (117% increase) and aged 75-59 (133% increase), among those of working age for those aged 60–64 years (91% increase).
Conclusion: Population-adjusted admissions for spondylodiscitis in England have risen by 44% between 2012 and 2021. Healthcare policymakers and providers must acknowledge the increasing burden of spondylodiscitis and make spondylodiscitis a research priority.
Oral e-Poster Presentations - Booth 2: Spine 1 (Trauma&Misc), September 25, 2023, 10:00 AM - 10:40 AM
Background: Spondylodiscitis is a prevalent type of spinal infection, with pyogenic spondylodiscitis being the most common subtype. While antibiotic therapy is the standard treatment, some argue that early surgery can aid in infection clearance, improve survival rates, and prevent long-term complications such as deformities. However, others view early surgery as excessively risky. Due to the high mortality rate of up to 20%, it is crucial to determine the most effective treatment.
Methods: The primary objective of this study was to compare the mortality rate, relapse rate, and length of hospital stay for conservative and early surgical treatments of pyogenic spondylodiscitis, including determinants of outcomes. The study was registered on PROSPERO with the registration number CRD42022312573. The databases MEDLINE, Embase, Scopus, PubMed, and JSTOR were searched for original studies comparing conservative and early surgical treatments of pyogenic spondylodiscitis. The included studies were assessed using the ROBINS-1 tool, and eligible studies were evaluated using meta-analyses, influence, and regression analyses.
Results: The systematic review included 31 studies. The meta-analysis, which had a pooled sample size of 10,954 patients from 21 studies, found that the pooled mortality rate among patients treated with early surgery was 8%, while the rate was 13% for patients treated conservatively. The mean proportion of relapse/failure was 15% for patients treated with early surgery and 21% for those treated conservatively. Furthermore, the analysis concluded that early surgical treatment is associated with a 40% and 39% risk reduction in relapse/failure and mortality rates, respectively, when compared to conservative management. Additionally, early surgical treatment resulted in a 7.75-day reduction in length of hospital stay per patient (p<0.01). The most highly significant predictors of treatment outcome were found to be intravenous drug use, diabetes, the presence of an epidural abscess, positive cultures, location of infection, and age (p<0.001).
Conclusions: Overall, early surgical management was found to be consistently significantly more effective than conservative management in terms of relapse/failure and mortality rates when treating pyogenic spondylodiscitis, particularly for non-spinal epidural abscess spondylodiscitis.
Introduction: Spondylodiscitis is the commonest form of infectious disease of the spine and harbours a high mortality rate of up to 20%. Recent demographic trends in Germany, such as an aging population, immunosuppression, and intravenous drug use, suggest that the incidence of spondylodiscitis may be on the rise. However, the exact epidemiological development of the disease remains uncertain. This study aims to analyse the burden on the tertiary healthcare system in Germany using data from the Federal Statistical Office of Germany (FSOG) database.
Materials and Methods: All cases of spondylodiscitis diagnosed between 2005 and 2021 were identified from the FSOG database. The study characterised the mean duration of hospital stays, total and population-adjusted number of diagnoses made, age-stratified incidence, and outcomes of hospitalised patients.
Results: A total of 131,982 diagnoses for spondylodiscitis were identified between 2005 and 2021. The number of diagnoses for spondylodiscitis has doubled during this period, from 5.4/100,000 population in 2005 to 11/100,000 population in 2021. The highest increase in admissions was recorded for those aged 90 years and above (+1307%), 80-89 (+376%) and 70-79 (+99%). Hospital discharges to rehabilitation facilities have increased by 160%, and discharges against medical advice by 91%. On the other hand, during the analysed period, the in-hospital mortality rate has decreased by 52%.
Conclusion: The population-adjusted incidence of spondylodiscitis in Germany has more than doubled between 2005 and 2021, highlighting the clinical relevance of this disease. During the same period, in-hospital mortality dropped by half. These findings suggest the need for further investigation into optimal therapy, particularly the role and timing of surgical treatment.
Investigators in the cognitive neurosciences have turned to Big Data to address persistent replication and reliability issues by increasing sample sizes, statistical power, and representativeness of data. While there is tremendous potential to advance science through open data sharing, these efforts unveil a host of new questions about how to integrate data arising from distinct sources and instruments. We focus on the most frequently assessed area of cognition - memory testing - and demonstrate a process for reliable data harmonization across three common measures. We aggregated raw data from 53 studies from around the world which measured at least one of three distinct verbal learning tasks, totaling N = 10,505 healthy and brain-injured individuals. A mega analysis was conducted using empirical bayes harmonization to isolate and remove site effects, followed by linear models which adjusted for common covariates. After corrections, a continuous item response theory (IRT) model estimated each individual subject’s latent verbal learning ability while accounting for item difficulties. Harmonization significantly reduced inter-site variance by 37% while preserving covariate effects. The effects of age, sex, and education on scores were found to be highly consistent across memory tests. IRT methods for equating scores across AVLTs agreed with held-out data of dually-administered tests, and these tools are made available for free online. This work demonstrates that large-scale data sharing and harmonization initiatives can offer opportunities to address reproducibility and integration challenges across the behavioral sciences.
Highlights
• Brain connectivity states identified by cofluctuation strength.
• CMEP as new method to robustly predict human traits from brain imaging data.
• Network-identifying connectivity ‘events’ are not predictive of cognitive ability.
• Sixteen temporally independent fMRI time frames allow for significant prediction.
• Neuroimaging-based assessment of cognitive ability requires sufficient scan lengths.
Abstract
Human functional brain connectivity can be temporally decomposed into states of high and low cofluctuation, defined as coactivation of brain regions over time. Rare states of particularly high cofluctuation have been shown to reflect fundamentals of intrinsic functional network architecture and to be highly subject-specific. However, it is unclear whether such network-defining states also contribute to individual variations in cognitive abilities – which strongly rely on the interactions among distributed brain regions. By introducing CMEP, a new eigenvector-based prediction framework, we show that as few as 16 temporally separated time frames (< 1.5% of 10 min resting-state fMRI) can significantly predict individual differences in intelligence (N = 263, p < .001). Against previous expectations, individual's network-defining time frames of particularly high cofluctuation do not predict intelligence. Multiple functional brain networks contribute to the prediction, and all results replicate in an independent sample (N = 831). Our results suggest that although fundamentals of person-specific functional connectomes can be derived from few time frames of highest connectivity, temporally distributed information is necessary to extract information about cognitive abilities. This information is not restricted to specific connectivity states, like network-defining high-cofluctuation states, but rather reflected across the entire length of the brain connectivity time series.
Objectives: To analyse carbapenemases in Proteus mirabilis and assess the performance of carbapenemase detection assays.
Methods: Eighty-one clinical P. mirabilis isolates with high-level resistance at least to ampicillin (>32 mg/L) or previous detection of carbapenemases were selected and investigated by three susceptibility testing methods (microdilution, automated susceptibility testing, and disk diffusion), six phenotypic carbapenemase assays (CARBA NP, modified carbapenemase inactivation method [CIM], modified zinc-supplemented CIM, simplified CIM, faropenem, and carbapenem-containing agar), two immunochromatographic assays, and whole-genome sequencing.
Results: Carbapenemases were detected in 43 of 81 isolates (OXA-48-like [n = 13]; OXA-23 [n = 12]; OXA-58 [n = 12]; New Delhi metallo-β-lactamase (NDM) [n = 2]; Verona integron–encoded metallo-β-lactamase (VIM) [n = 2]; Imipenemase (IMP) [n = 1]; Klebsiella pneumoniae carbapenemase (KPC) [n = 1]). Carbapenemase-producing Proteus were frequently susceptible to ertapenem (26/43; 60%), meropenem (28/43; 65%), ceftazidime (33/43; 77%), and some even to piperacillin-tazobactam (9/43; 21%). Sensitivity/specificity of phenotypic tests were 30% (CI: 17–46%)/89% (CI: 75–97%) for CARBA NP, 74% (CI: 60–85%)/82% (CI: 67–91%) for faropenem, 91% (CI: 78–97%)/82% (CI: 66–92%) for simplified CIM, and 93% (CI: 81–99%)/100% (CI: 91–100%) for modified zinc-supplemented CIM. An algorithm for improved detection was developed, which demonstrated sensitivity/specificity of 100% (CI: 92–100%)/100% (CI: 91–100%) on the 81 isolates, and 100% (CI: 29–100%)/100% (CI: 96–100%) in a prospective analysis of additional 91 isolates. Interestingly, several OXA-23-producing isolates belonged to the same clonal lineage reported previously from France.
Discussion: Current susceptibility testing methods and phenotypic tests frequently fail to detect carbapenemases in P. mirabilis, which could result in inadequate antibiotic treatment. In addition, the non-inclusion of blaOXA-23/OXA-58 in many molecular carbapenemase assays further impedes their detection. Therefore, the prevalence of carbapenemases in P. mirabilis is likely underestimated. With the herein proposed algorithm, carbapenemase-producing Proteus can be easily identified.
Chromosomal translocations (CTs) are a genetic hallmark of cancer. They could be identified as recurrent genetic aberrations in hemato-malignancies and solid tumors. More than 40% of all “cancer genes” were identified in recurrent CTs. Most of these CTs result in the production of oncofusion proteins of which many have been studied over the past decades. They influence signaling pathways and/or alter gene expression. However, a precise mechanism for how these CTs arise and occur in a nearly identical fashion in individuals remains to be elucidated. Here, we performed experiments that explain the onset of CTs: (1) proximity of genes able to produce prematurely terminated transcripts, which lead to the production of (2) trans-spliced fusion RNAs, and finally, the induction of (3) DNA double-strand breaks which are subsequently repaired via EJ repair pathways. Under these conditions, balanced chromosomal translocations could be specifically induced. The implications of these findings will be discussed.
IL-38 is the latest discovered cytokine of the IL-1 family and has been added to the IL-36 subfamily. Since its discovery in 2001, increasing evidence suggests predominantly anti-inflammatory properties of IL-38, which are most likely exerted through three potential receptors, the IL-1 Receptor 1 (IL-1R1), IL-36 Receptor (IL-36R) and the IL-1 Receptor Accessory Protein Like 1 (IL-1RAPL1). However, to this date detailed knowledge of IL-38 functioning remains to be examined. Importantly, how IL-38 is processed, secreted from cells and the exact mechanisms of target receptor binding and intracellular signaling are not fully understood. Further, IL-38 has been associated with regulatory functions in autoimmune diseases like systemic lupus erythematosus (SLE) and psoriasis. At the same time however, connections between B cells as indispensable part of immunity and IL-38 remain rare.
In this study we examined the influence of IL-38 in peripheral human blood B cells differentiating into antibody secreting cells using a three-step in vitro differentiation process. We first show that all potential IL-38 binding receptors are present on peripheral blood B cells on a gene expression level and remain detectable throughout B cell differentiation. Next, while B cells treated with exogenous IL-38 depict no differences in early B cell activation markers, the process of B cell differentiation revealed significant alterations in B cell phenotype created by IL-38 treatment. Predominantly on day 7 of the differentiation process, IL-38 treated B cells showed significantly reduced CD38 expression which depicts an important step in development towards plasma cells. We hypothesize that IL-38 acts antagonistically on the IL-1R1 pathway reducing Nuclear factor kappa B (NFκB) expression and consequently decreasing CD38 expression. Further IL-38 reduced early antibody production while increasing IgM secretion at the end stages of differentiation. Next, we repeated the differentiation assays under the influence of additional IL-21 stimulation to further enhance plasma cell development. In these experiments, the impact of IL-38 on B cell differentiation and immunoglobulin production were reduced, indicating a comparatively moderate relevance of IL-38 for B cell differentiation. We then examined how proliferation and cell death were impacted by exogenous IL-38 during B cell differentiation. IL-38 treatment alone significantly reduced B cell survival which was further augmented by IL-21 stimulation. We conclude that IL-38 and IL-21 act synergistically in promoting B cell apoptosis, also depicting an anti-inflammatory property of IL-38. Finally, using a siRNA we successfully performed an IL-38 knockdown experiment of human blood B cells reducing IL-38 expression to 44% measured on day 4 of B cell differentiation. In these experiments we observed reversed tendencies of CD38 expression compared to exogenous IL-38 treatment. Here, IL-38 knockdown cells showed increased CD38 expression indicating endogenous regulatory properties of IL-38 in B cell differentiation.
Our project, for the first time proves direct effects of IL-38 on human B cells. The results support previous research of IL-38 to act anti-inflammatory as it seems to modulate B cell differentiation, survival, and immunoglobulin production in a down-regulatory manner. These findings pave way for more detailed research on the connection between B cell homoeostasis and IL-38 function.
Refraktives und visuelles Ergebnis nach Neuausrichtung von rotierten torischen Intraokularlinsen
(2023)
Hintergrund und Ziel der Studie
Das Ziel der Studie ist die Beurteilung des refraktiven und visuellen Ergebnisses von Patienten mit fehlausgerichteten torischen IOLs nach operativer Neuausrichtung, mit und ohne Rückberechnung der torischen Achse nach Implantation der IOL.
Methoden
Dies ist eine retrospektive Analyse von 39 Patienten, die sich von August 2013 bis Dezember 2019 an der Klinik für Augenheilkunde der Goethe-Universität Frankfurt am Main einer zweiten Operation zur Neuausrichtung einer fehlausgerichteten torischen IOL unterzogen haben. Die zweite Operaton zur Korrektur der Achsenlage wurde auf zwei Weisen durchgeführt: In der ersten Gruppe wurde auf die primär berechnete Achse rotiert und in der zweiten Gruppe wurde die Rotation auf eine neu berechnete Achse mit dem Rückrechner von astigmatisfix.com vorgenommen.
Ergebnisse
In dieser retrospektiven Studie wurden 39 Augen von 39 Patienten eingeschlossen. Die Ergebnisse nach der ersten Operation (die Linsenoperation) zeigte eine Besserung der (BCVA) von 0,28 ± 0,22 logMAR (20/40) auf 0,15 ± 0,14 logMAR (20/32). Nach der Linsenoperation zeigten die torischen IOLs durchschnittlich eine postoperative Fehlstellung von 25,69° ± 26,06°. Verglichen vor und nach der Rotation der torischen IOL blieb der BCVA unverändert bei 0,14 ± 0,14 logMAR (20/32), während sich der UDVA signifikant von 0,39 ± 0,29 logMAR (20/50) auf 0,27 ± 0,18 logMAR (20/40) verbesserte. Das refraktive Ergebnis zeigte eine Reduktion sowohl in der Restsphäre sowie in dem Zylinder. Die Ergebnisse der Gruppenanalyse demonstrierte für die erste Gruppe (Neuausrichtung auf die präoperativ berechnete Achse) eine postoperative UDVA 0,24 ± 0,16 logMAR mit einem Zylinder von 0,90 ± 0,90 D. In der zweiten Gruppe (Ausrichtung auf eine zurückberechnete Achse) betrug die UDVA 0,32 ± 0,20 logMAR mit einem Zylinder von 0,76 ± 0,72 D. Die IOLs mit hoher Zylinderstärke (≥ 2 D) zeigten bei der Rückberechnung eine stärkere Abnahme des Restzylinders als IOL mit niedriger Zylinderstärke (< 2 D) (27 % vs. 9 %). Der mittlere sphärische Äquivalent-Vorhersagefehler der Rückberechnung betrug 0,54 ± 0,55 D.
Schlussfolgerung
Die Neuausrichtung von fehlausgerichteten torischen IOLs verbessert die Sehschärfe und reduziert verbleibende Brechungsfehler. Insbesondere bei IOL mit hoher Zylinderstärke kann ein besseres refraktives Ergebnis erzielt werden, wenn vor der Neuausrichtung eine Rückberechnung durchgeführt wird.
Epigenetic neural glioblastoma enhances synaptic integration and predicts therapeutic vulnerability
(2023)
Neural-tumor interactions drive glioma growth as evidenced in preclinical models, but clinical validation is nascent. We present an epigenetically defined neural signature of glioblastoma that independently affects patients survival. We use reference signatures of neural cells to deconvolve tumor DNA and classify samples into low- or high-neural tumors. High-neural glioblastomas exhibit hypomethylated CpG sites and upregulation of genes associated with synaptic integration. Single-cell transcriptomic analysis reveals high abundance of stem cell-like malignant cells classified as oligodendrocyte precursor and neural precursor cell-like in high-neural glioblastoma. High-neural glioblastoma cells engender neuron-to-glioma synapse formation in vitro and in vivo and show an unfavorable survival after xenografting. In patients, a high-neural signature associates with decreased survival as well as increased functional connectivity and can be detected via DNA analytes and brain-derived neurotrophic factor in plasma. Our study presents an epigenetically defined malignant neural signature in high-grade gliomas that is prognostically relevant.
Compressive knee joint contact force during walking is thought to be related to initiation and progression of knee osteoarthritis. However, joint loading is often evaluated with surrogate measures, like the external knee adduction moment, due to the complexity of computing joint contact forces. Statistical models have shown promising correlations between medial knee joint contact forces and knee adduction moments in particularly in individuals with knee osteoarthritis or after total knee replacements (R2 = 0.44–0.60). The purpose of this study was to evaluate how accurately model-based predictions of peak medial and lateral knee joint contact forces during walking could be estimated by linear mixed-effects models including joint moments for children and adolescents with and without valgus malalignment. Peak knee joint moments were strongly correlated (R2 > 0.85, p < 0.001) with both peak medial and lateral knee joint contact forces. The knee flexion and adduction moments were significant covariates in the models, strengthening the understanding of the statistical relationship between both moments and medial and lateral knee joint contact forces. In the future, these models could be used to evaluate peak knee joint contact forces from musculoskeletal simulations using peak joint moments from motion capture software, obviating the need for time-consuming musculoskeletal simulations.
Die rechtsmedizinische Begutachtung von Knochenfunden : eine hessenweite retrospektive Studie
(2023)
Einleitung: Zufällige Funde von Knochen(fragmenten) können von großem Interesse für z. B. Strafverfolgungs- und Denkmalbehörden oder Bauträger sein. Zur Beurteilung einer etwaigen strafrechtlichen oder historischen Relevanz tragen forensisch-osteologische Untersuchungen in einem hohen Maße bei. Die vorliegende Arbeit soll einen Überblick über die forensisch-osteologische Arbeit der beiden hessischen Institute für Rechtsmedizin geben und durch die Interpretation der Ergebnisse im (inter-)nationalen Vergleich regionale Besonderheiten und die Rolle der Rechtsmedizin bei der Begutachtung aufzeigen. Der daraus entstandene Erkenntnisgewinn soll ein Problembewusstsein schaffen sowie Fallarbeit und Lehre verbessern.
Material und Methoden: Zunächst wurden für den Zeitraum von 2005 bis 2019 die im Institut für Rechtsmedizin Gießen archivierten osteologischen Gutachten deskriptiv statistisch ausgewertet. Es wurden Daten zu Fundort, -umständen, Humanspezifität, postmortalem Intervall, Spuren von Gewalt und die Ergebnisse weiterführender Untersuchungen erhoben.
In einer zweiten Studie wurde die Auswertung auf archivierte forensisch- osteologische Gutachten des Instituts für Rechtsmedizin Frankfurt am Main ausgeweitet und der Auswertungszeitraum auf die Jahre 2011 bis einschließlich 2020 angepasst, um eine hessenweite Datengrundlage zu generieren.
Ergebnisse: In der ersten Studie wurden von den in dem 15-Jahres-Zeitraum begutachteten 172 Knochenfunden aus dem Institut für Rechtsmedizin Gießen 40 % in Wald- und Wiesengebieten aufgefunden. In 58 % enthielten die Funde menschliche Knochen, davon wurde in 32 % eine forensisch relevante Liegezeit von 50 Jahren oder weniger nicht ausgeschlossen, und 6% wiesen Zeichen perimortaler Gewalteinwirkung auf. Bei den Fällen, in denen eine Humanspezifität ausgeschlossen wurde, handelte es sich am häufigsten um Knochen von Hirsch bzw. Reh (32 %), Schwein (29 %) und Rind (14 %). In 20 % aller Fälle wurden
ergänzende DNA-Untersuchungen durchgeführt; in 62 % verlief die Typisierung humaner STR-Systeme erfolgreich und davon gelang in 41 % die Zuordnung zu einem antemortalen Profil einer vermissten Person.
In der zweiten, hessenweiten Studie, wurden 288 Knochenfunde eines 10-Jahres-Zeitraums ausgewertet. Mit 38,2 % wurden die meisten Funde in Wäldern, Wiesen und großen Parks getätigt. In 50,7 % der Fälle enthielten die Funde menschliche Knochen, davon wurde in 37,0 % eine forensisch relevante Liegezeit angenommen. Zeichen einer Gewalteinwirkung wurden in 77,4 % der Fälle mit menschlichen Knochen beschrieben, dabei handelte es sich mit 78,8 % am häufigsten um postmortale Beschädigungen, in 9,7 % um perimortale und in 11,5 % um antemortale Verletzungen. In 40,4 % der als human klassifizierten Knochenfunde wurden DNA-Untersuchungen durchgeführt. Dabei konnte in 81,3 % ein STR-Profil erstellt werden, das in 35,4% zu einer sicheren Identifizierung führte. Bei den zur Untersuchung überstellten nicht-menschlichen Knochen handelte es sich am häufigsten um Knochen vom Schwein (23,4 %), Hirsch (18,1 %), Rind (16,4 %), Reh (11,7 %) und Schaf (11,7 %).
Diskussion: Die Begutachtung von Knochen und Knochenfragmenten ist ein fester Bestandteil der rechtsmedizinischen Fallarbeit und zwingend erforderlich, um Fälle von forensischem Interesse zu identifizieren. Die makroskopische Befundung durch einen geschulten Untersucher erlaubt in der Regel bereits, Aussagen über die forensische Relevanz eines Fundes zu treffen und so die Weichen für das weitere Vorgehen zu stellen. Hilfreich bei der makroskopischen Beurteilung ist eine gute Kenntnis der regionalen Besonderheiten in dem Gebiet, in dem die Knochen gefunden wurden. Die makroskopische forensisch-osteologische Begutachtung stellt trotz der sich ständig weiterentwickelnden technischen Möglichkeiten ein unverzichtbares Werkzeug in der Bearbeitung von Knochenfunden dar. In Deutschland ist es Aufgabe der Rechtsmedizin, diese Expertise zu pflegen und vorzuhalten, mit dem Ziel, zur Aufklärung von Vermisstenfällen beizutragen und unentdeckte nichtnatürliche Todesfälle aufzuklären.
Highlights
• Early reconstruction of injured cruciate ligaments improves functional outcomes.
• Modern CT imaging can be used to rapidly identify patients with injury to the cruciate ligaments and streamline therapeutic pathways.
• Dual-energy CT demonstrates superior diagnostic accuracy compared to single-energy CT.
Abstract
Background: This study aimed to evaluate the clinical utility of modern single and dual-energy computed tomography (CT) for assessing the integrity of the cruciate ligaments in patients that sustained acute trauma.
Methods: Patients who underwent single- or dual-energy CT followed by 3 Tesla magnetic resonance imaging (MRI) or knee joint arthroscopy between 01/2016 and 12/2022 were included in this retrospective, monocentric study. Three radiologists specialized in musculoskeletal imaging independently evaluated all CT images for the presence of injury to the cruciate ligaments. An MRI consensus reading of two experienced readers and arthroscopy provided the reference standard. Diagnostic accuracy parameters and area under the receiver operator characteristic curve (AUC) were the primary metrics for diagnostic performance.
Results: CT images of 204 patients (median age, 49 years; IQR 36 – 64; 113 males) were evaluated. Dual-energy CT yielded significantly higher diagnostic accuracy and AUC for the detection of injury to the anterior (94% [240/255] vs 75% [266/357] and 0.89 vs 0.66) and posterior cruciate ligaments (95% [243/255] vs 87% [311/357] and 0.90 vs 0.61) compared to single-energy CT (all parameters, p <.005). Diagnostic confidence and image quality were significantly higher in dual-energy CT compared to single-energy CT (all parameters, p <.005).
Conclusions: Modern dual-energy CT is readily available and can serve as a screening tool for detecting or excluding cruciate ligament injuries in patients with acute trauma. Accurate diagnosis of cruciate ligament injuries is crucial to prevent adverse outcomes, including delayed treatment, chronic instability, or long-term functional limitations.
Purpose: To evaluate intermediate and long-term visual outcomes and safety of a phakic intraocular posterior chamber lens with a central hole (ICL V4c) for myopic eyes.
Methods: Retrospective, consecutive case study of patients that uneventfully received a ICL V4c for myopia correction, with a 5-year postoperative follow-up. Department of Ophthalmology, Goethe University Frankfurt, Germany.
Results: From 241 eyes that underwent ICL implantation, we included 45 eyes with a mean age at surgery of 33 years ± 6 (18–48 years), with a 5 years follow-up. CDVA improved from 0.05logMAR ± 0.15 CDVA preoperatively to − 0.00 ± 0,07 at 5 years and did not change significantly from 3 to 5 years’ time (p = 0.266). The mean spherical equivalent (SE) improved from -10.13D ± 3.39 to − 0.45D ± 0.69. The change in endothelial cell count showed a mean decrease of 1.9% per year throughout the follow-up. Safety and efficacy index were 1.16 and 0.78, respectively. Cataract formation was seen in 2 of 241 eyes (0.8%), but in none of the 45 eyes that finished the 5-year follow-up.
Conclusions: Our data show a good intermediate and long-term stability, efficiency, and safety of ICL V4c phakic lenses in myopic eyes comparable to other known literature.
Background/aim: The aim of this study was to analyze a population of patients who had suffered from traumatic dental injuries (TDIs) by using different patient-, trauma- and treatment-related parameters.
Material and methods: All dental records of patients ≥ 3 years old who had presented at the dental emergency service between Jan 1, 2009 and Dec 31, 2016 for the treatment of dental trauma were analyzed. A total of 2758 patients were invited for a recall examination at the Department for Dental Surgery and Implantology, ZZMK Carolinum, Goethe University Frankfurt, Germany; of these, 269 patients attended their recall appointments.
Results: The enrolled patient population consisted of 1718 males and 1040 females, with a mean age of 19.63 years (median 12.00 ± 17.354 years). A total of 4909 injured teeth were assessed, with a mean of 1.78 injured teeth per patient (median 2.00 ± 1.279). Males were found to be more frequently affected by TDIs compared to females (1.65:1). The majority of these injuries occurred in the first two decades of life (66.1%; n = 1824). The majority of the patients presented for initial treatment within 24 h of their accident (95.7%). The most frequent TDIs were isolated luxation injuries 49.4% (n = 2426) and isolated crown fractures 30% (n = 1472). Combination injuries were diagnosed in 20.6% of the cases (n = 1011).
Conclusions: Based on the findings of the present analysis, it can be concluded that males were more frequently affected by TDIs than females. Most patients had suffered from TDI before they had turned 10 years of age. Overall, the enamel–dentin fracture was found to be the most frequent injury, followed by concussions and lateral luxations.
MicroRNAs (miRNAs) are critical post-transcriptional regulators in many biological processes. They act by guiding RNA-induced silencing complexes to miRNA response elements (MREs) in target mRNAs, inducing translational inhibition and/or mRNA degradation. Functional MREs are expected to predominantly occur in the 3' untranslated region and involve perfect base-pairing of the miRNA seed. Here, we generate a high-resolution map of miR-181a/b-1 (miR-181) MREs to define the targeting rules of miR-181 in developing murine T-cells. By combining a multi-omics approach with computational high-resolution analyses, we uncover novel miR-181 targets and demonstrate that miR-181 acts predominantly through RNA destabilization. Importantly, we discover an alternative seed match and identify a distinct set of targets with repeat elements in the coding sequence which are targeted by miR-181 and mediate translational inhibition. In conclusion, deep profiling of MREs in primary cells is critical to expand physiologically relevant targetomes and establish context-dependent miRNA targeting rules.
Background: Epilepsy surgery is an established treatment for drug-resistant focal epilepsy (DRFE) that results in seizure freedom in about 60% of patients. Correctly identifying an epileptogenic lesion in magnetic resonance imaging (MRI) is challenging but highly relevant since it improves the likelihood of being referred for presurgical diagnosis. The epileptogenic lesion’s etiology directly relates to the surgical intervention’s indication and outcome. Therefore, it is vital to correctly identify epileptogenic lesions and their etiology presurgically.
Methods: We compared the final histopathological diagnoses of all patients with DRFE undergoing epilepsy surgery at our center between 2015 and 2021 with their MRI diagnoses before and after presurgical diagnosis at our epilepsy center, including MRI evaluations by expert epilepsy neuroradiologists. Additionally, we analyzed the outcome of different subgroups.
Results: This study included 132 patients. The discordance between histopathology and MRI diagnoses significantly decreased from 61.3% for non-expert MRI evaluations (NEMRIs) to 22.1% for epilepsy center MRI evaluations (ECMRIs; p < 0.0001). The MRI-sensitivity improved significantly from 68.6% for NEMRIs to 97.7% for ECMRIs (p < 0.0001). Identifying focal cortical dysplasia (FCD) and amygdala dysplasia was the most challenging for both subgroups. 65.5% of patients with negative NEMRI were seizure-free 12 months postoperatively, no patient with negative ECMRI achieved seizure-freedom. The mean duration of epilepsy until surgical intervention was 13.6 years in patients with an initial negative NEMRI and 9.5 years in patients with a recognized lesion in NEMRI.
Conclusions: This study provides evidence that for patients with DRFE—especially those with initial negative findings in a non-expert MRI—an early consultation at an epilepsy center, including an ECMRI, is important for identifying candidates for epilepsy surgery. NEMRI-negative findings preoperatively do not preclude seizure freedom postoperatively. Therefore, patients with DRFE that remain MRI-negative after initial NEMRI should be referred to an epilepsy center for presurgical evaluation. Nonreferral based on NEMRI negativity may harm such patients and delay surgical intervention. However, ECMRI-negative patients have a reduced chance of becoming seizure-free after epilepsy surgery. Further improvements in MRI technique and evaluation are needed and should be directed towards improving sensitivity for FCDs and amygdala dysplasias.
A broad range of neuropsychiatric disorders are associated with alterations in macroscale brain circuitry and connectivity. Identifying consistent brain patterns underlying these disorders by means of structural and functional MRI has proven challenging, partly due to the vast number of tests required to examine the entire brain, which can lead to an increase in missed findings. In this study, we propose polyconnectomic score (PCS) as a metric designed to quantify the presence of disease-related brain connectivity signatures in connectomes. PCS summarizes evidence of brain patterns related to a phenotype across the entire landscape of brain connectivity into a subject-level score. We evaluated PCS across four brain disorders (autism spectrum disorder, schizophrenia, attention deficit hyperactivity disorder, and Alzheimer’s disease) and 14 studies encompassing ∼35,000 individuals. Our findings consistently show that patients exhibit significantly higher PCS compared to controls, with effect sizes that go beyond other single MRI metrics ([min, max]: Cohen’s d = [0.30, 0.87], AUC = [0.58, 0.73]). We further demonstrate that PCS serves as a valuable tool for stratifying individuals, for example within the psychosis continuum, distinguishing patients with schizophrenia from their first-degree relatives (d = 0.42, p = 4 x 10−3, FDR-corrected), and first-degree relatives from healthy controls (d = 0.34, p = 0.034, FDR-corrected). We also show that PCS is useful to uncover associations between brain connectivity patterns related to neuropsychiatric disorders and mental health, psychosocial factors, and body measurements.
Background: Multiple studies have focused on medical and pharmacological treatments and outcome predictors of patients with status epilepticus (SE). However, a sufficient understanding of recurrent episodes of SE is lacking. Therefore, we reviewed recurrent SE episodes to investigate their clinical characteristics and outcomes in patients with relapses.
Methods: In this retrospective, multicenter study, we reviewed recurrent SE patient data covering 2011 to 2017 from the university hospitals of Frankfurt and Marburg, Germany. Clinical characteristics and outcome variables were compared among the first and subsequent SE episodes using a standardized form for data collection.
Results: We identified 120 recurrent SE episodes in 80 patients (10.2% of all 1177 episodes). The mean age at the first SE episode was 62.2 years (median 66.5; SD 19.3; range 21–91), and 42 of these patients were male (52.5%). A mean of 262.4 days passed between the first and the second episode. Tonic–clonic seizure semiology and a cerebrovascular disease etiology were predominant in initial and recurrent episodes. After subsequent episodes, patients showed increased disability as indicated by the modified Rankin Scale (mRS), and 9 out of 80 patients died during the second episode (11.3%). Increases in refractory and super-refractory SE (RSE and SRSE, respectively) were noted during the second episode, and the occurrence of a non-refractory SE (NRSE) during the first SE episode did not necessarily provide a protective marker for subsequent non-refractory episodes. An increase in the use of intravenous-available anti-seizure medication (ASM) was observed in the treatment of SE patients. Patients were discharged from hospital with a mean of 2.8 ± 1.0 ASMs after the second SE episode and 2.1 ± 1.2 ASMs after the first episode. Levetiracetam was the most common ASM used before admission and on discharge for SE patients.
Conclusions: This retrospective, multicenter study used the mRS to demonstrate worsened outcomes of patients at consecutive SE episodes. ASM accumulations after subsequent SE episodes were registered over the study period. The study results underline the necessity for improved clinical follow-ups and outpatient care to reduce the health care burden from recurrent SE episodes.
Rationale and objectives: To provide a detailed analysis of injury patterns of the spine following blunt trauma and establish the role of supplementary MRI by evaluating discrepancies in the detection rates of damaged structures in CT and MRI.
Method: 216 patients with blunt trauma to the spine who underwent CT followed by supplementary MRI were included in this study. Two board-certified radiologists blinded to clinical symptoms and injury mechanisms independently interpreted all acquired CT and MRI images. The interpretation was performed using a dedicated catalogue of typical findings associated with spinal trauma and assessed for spinal stability using the AO classification systems.
Results: Lesions to structures associated with spinal instability were present in 31.0% in the cervical spine, 12.3% in the thoracic spine, and 29.9% in the lumbar spine. In all spinal segments, MRI provided additional information regarding potentially unstable injuries. Novel information derived from supplementary MRI changed clinical management in 3.6% of patients with injury to the cervical spine. No change in clinical management resulted from novel information on the thoracolumbar spine. Patients with injuries to the vertebral body, intervertebral disc, or spinous process were significantly more likely to benefit from supplementary MRI.
Conclusion: In patients that sustained blunt spinal trauma, supplementary MRI of the cervical spine should routinely be performed to detect injuries that require surgical treatment, whereas CT is the superior imaging modality for the detection of unstable injuries in the thoracolumbar spine.
Ein intaktes Arbeitsgedächtnis ermöglicht es, Informationen, z.B. visuellen oder auditorischen Ursprungs temporär zu speichern, zu manipulieren und weiterzuverarbeiten. Es bildet die Grundlage zum Lernen, zum Begreifen und zum Verstehen von Sprache und somit die Grundlage zum tagtäglichen Funktionieren.
Beeinträchtigungen des Arbeitsgedächtnisses sind mit schlechten (Arbeits-) Leistungen, eingeschränkter zwischenmenschlicher Kommunikation, niedrigerem gesellschaftlichem Funktionsniveau und Arbeitslosigkeit vergesellschaftet und somit folgenschwer.
Psychiatrische Erkrankungen aus dem schizophrenen Spektrum, die unipolare Depression oder das Aufmerksamkeitsdefizit-Hyperaktivitätssyndrom (ADHS) gehen gehäuft mit kognitiven Beeinträchtigungen einher. Auch die bipolare affektive Störung zählt zu den psychiatrischen Erkrankungen, die mit kognitiver Beeinträchtigung assoziiert ist.
Das Ziel der Studie war es, darzustellen, dass kognitive Einschränkungen von Patienten, die an einer bipolaren affektiven Störung leiden, unter den exekutiven Funktionen insbesondere das Arbeitsgedächtnis betreffen und diese auch in euthymer Stimmungslage zu erfassen sind.
Hierzu wurde mittels einer Aufgabe zur Farbveränderung vier verschiedener Kreise, die um ein zentral platziertes, schwarzes Kreuz angeordnet waren, das visuelle Arbeitsgedächtnis anhand der Merkfähigkeit der Patienten, deren euthyme Stimmungslage mit den Ratingskalen MADRS und YMRS bestätigt wurde, im Vergleich zu einer Gruppe gesunder Kontrollprobanden, die mit der Patientengruppe in den Faktoren Alter, Geschlechterverteilung, prämorbide Intelligenz, Schulbildung und Händigkeit vergleichbar war, überprüft.
Anhand der erzielten Ergebnisse wurde die Arbeitsgedächtniskapazität K nach Pashler bestimmt. Es ergab sich eine bei mittlerer Effektstärke signifikant geringere Arbeitsgedächtniskapazität in der Gruppe der remittierten Patienten mit bipolarer affektiver Störung im Vergleich zur Kontrollgruppe gesunder Probanden.
Ein Einfluss der zum Zeitpunkt der Testung in der Patientengruppe angewandten Psychopharmaka auf die Arbeitsgedächtniskapazität konnte ausgeschlossen werden, sodass die ermittelten Defizite nicht als eine Form von Medikamentennebenwirkungen erklärt werden konnten. Interessanterweise betrug die mediane Erkrankungsdauer in der Patientengruppe drei Jahre, sodass davon auszugehen ist, dass die Ausprägung von Defiziten des Arbeitsgedächtnisses keinen langen Krankheitsverlauf bedingt, sondern bereits zu Beginn der Erkrankung vorhanden sein könnte.
Somit konnte die Studie darlegen, dass kognitive Defizite, insbesondere Störungen des Arbeitsgedächtnisses mit daraus resultierender signifikant erniedrigter Arbeitsgedächtniskapazität, bei Patienten mit bipolarer affektiver Störung in euthymer Stimmungslage bereits zu einem frühen Zeitpunkt des Krankheitsverlaufs bestehen, die eine Folge der Erkrankung selbst sind und keine Nebenwirkung von psychopharmazeutischer Medikation darstellen. Den Grund für das Auftreten kognitiver Defizite bei Patienten mit bipolarer affektiver Störung liefert diese Studie nicht, sie impliziert dennoch, dass das Verständnis und die Berücksichtigung kognitiver Störungen bei der bipolaren affektiven Störung zukünftig ein wichtiges Therapieziel darstellen sollte um das funktionelle Outcome von Patienten, die an dieser Störung leiden, entscheidend zu verbessern und somit eine Teilnahme am tagtäglichen Funktionieren weiterhin ermöglicht wird.
In die Zukunft blickend sollten unsere Ergebnisse mithilfe weiterer Untersuchungen validiert werden, insbesondere in Bezug auf genetische Komponenten, die die Arbeitsgedächtniskapazität regulieren. Es sollten Faktoren identifiziert werden, die eine reduzierte Arbeitsgedächtniskapazität bei Patienten mit bipolarer affektiver Störung bedingen. Die vorliegende Studie beantwortet nämlich nicht die Frage, wie genetische Faktoren und Umweltfaktoren die Arbeitsgedächtniskapazität auf kognitiver und neurophysiologischer Ebene beeinflussen könnten.
Angesichts der erwähnten Relevanz eines intakten Arbeitsgedächtnisses für die Funktionsfähigkeit der Patienten sollten zukünftige Studien prüfen, ob prokognitive Interventionen wie die kognitive Remediation oder sogar medikamentöse Therapieoptionen diese Defizite verbessern könnten.
Metastatic rhabdomyosarcoma (RMS) is one of the most challenging tumor entities in pediatric oncology caused by treatment resistances and immune escape. Novel chimeric antigen receptor (CAR) immunotherapies as specific, effective and safe treatment provide antitumor cytotoxicity by soluble factors and ligands/receptor signals. Besides its intrinsic potential as innate immune cell the ErbB2-sprecific CAR-engineered natural killer (NK)-92 cell line NK-92/5.28.z also provides CAR-mediated cytotoxicity, resulting in a high lytic capacity against 2D and 3D RMS cell structures in vitro. Also in a xenograft model using immune deficient NOD/Scid/IL2Rγ-/- (NSG) mice inhibited NK-92/5.28.z the tumor growth as long as the cells were administered and therefore prolonged the survival of the animals. The NK-92/5.28.z were distributed by the blood circulation and subsequently infiltrated the tumor tissue. Due to the malignant origin of the NK-92 cell line the cells must be irradiated prior to the use in patients. While the irradiation hampered the proliferation of NK-92/5.28.z cells, the cytotoxicity against RMS cells in vitro is retained for at least 24 hours. In the xenograft model irradiated NK-92/5.28.z cells inhibited the tumor growth but to a lower extent than untreated cells, as irradiated cells have only a limited life span in vivo no durable persistence and remission was achieved. Therefore, combinatorial approaches were focused and while blocking of the PD-1/PD-L1 axis did not resulted in a significantly enhanced tumor cell lysis, the combinatorial treatment with proteasome inhibitor bortezomib exhibited a significant enhanced cytotoxicity against RMS cells at least in vitro. Bortezomib itself induces caspase mediated apoptosis and also the upregulates the expression of TRAIL receptor DR5. The corresponding ligand TRAIL is expressed on the surface of the NK-92/5.28.z and pursuing experiments with purified TRAIL and bortezomib revealed a synergism. NK-92/5.28.z as an off-the-shelf product is therefore feasible for the therapy of metastatic RMS, but it might be necessary to support the cytotoxicity by additive agents like proteasome inhibitor bortezomib to archive durable remission.
Another cell population suitable for RMS CAR-immunotherapy are cytokine induced killer (CIK) cells, a heterogenous cell population generated from autologous PBMCs consisting of T, NK and T-NK cells. Lentivirally transduced ErbB2-specific CAR-CIK cells were previously shown to inhibit the tumor engraftment in a RMS xenograft model. However, lentiviral transduced adoptive immunotherapies bear risks for the transfer in patients, therefore the Sleeping Beauty Transposon System (SBTS) as a non-viral method, which integrates the CAR coding DNA by a cut-and-paste mechanism from a minicircle (MC) into the CIK cells genome is more feasible for the generation of CAR-CIK cells. The Sleeping beauty transposase mRNA and the MC were transferred in the cell by nucleofection, different factors influence the transfection efficiency and viability of the CIK cells in this harsh procedure. In preliminary experiments with MC Venus, a MC encoding eGFP, the highest transfection efficiency with the best proliferative capacity was achieved with cells on day 3 of CIK culture and without the addition of autologous monocytes as feeder cells. For the CAR construct the protocol was further improved by adjusting crucial factors, for this construct the best results were achieved on day 0, without irradiated PBMCs as feeder cells and cultivation in X-Vivo10 medium supplemented with human fresh frozen plasma. The X-Vivo10 medium enhanced the percentage of NK- and T-NK cells significantly compared to CAR-CIK cells cultured in RPMI. Since the gene transfer by SBTS resulted in CAR-CIK cells stably expressing a CAR in all subpopulations, resulting in a significantly enhanced cytotoxicity against RMS cells in vitro, these cells were compared to lentiviral transduced CAR-CIK cells in vitro and in vivo. While the SBTS CAR-CIK cells were superior to viral CAR-CIK cells in 2D short-term assays, the viral cells showed higher lytic capacity in 3D spheroid long-term assays. In a RMS xenograft model lentiviral CAR-CIK cells significantly prolonged the survival of mice and persisted, whereas SBTS CAR-CIKs did not favor the overall survival compared to untreated controls and also did not persist. Phenotypic analysis revealed a highly cytotoxic CD8+ and late effector memory dominant phenotype for SBTS CAR-CIK cells supporting short-term cytotoxicity but also more prone for exhaustion, while viral CAR-CIK cells showed a more balanced phenotype for memory and cytotoxicity. Therefore, the SBTS is feasible for the ErbB2-CAR gene transfer in CAR-CIK resulting in a stable CAR-expression with high short-term cytotoxicity, but these cells are also more prone to exhaustion and the protocol might be adapted further to prevent this limitation for in vivo application.
This work underlines the hard-to-treat characteristics of metastatic RMS, but also shows some approaches for further evaluation like the combination of NK-92/5.28.z cells with bortezomib and the feasibility of the generation of CAR-CIK cells via SBTS.
In dieser retrospektiven Studie wurden 47 Patienten mit einem histologisch nachgewiesenen Analkarzinom und im Anschluss aufgetretenen Rezidiv nach Beendigung der Radiochemotherapie und hinsichtlich der Risikofaktoren für das tumorfreie, sowie das Gesamtüberleben ausgewertet.
Auffällig in dieser Patientenkohorte war der hohe Anteil an männlichen Patienten (68,1 %), an HIV-positiven (36,2 %) sowie an Patienten mit nachgewiesenem Lymphknotenbefall bei Erstdiagnose (72,3 %).
Als signifikante Risikofaktoren für das tumorfreie Überleben wurden ein Primärtumor ab ≥ T3-Kategorie (Hazard-Ratio 1,87), ein Karnofsky-Performance-Status ≤ 80 % vor Beginn der Radiochemotherapie (Hazard-Ratio 3,25), sowie das initiale fehlende therapeutische Ansprechen der Radiochemotherapie (Hazard-Ratio 5,9) festgestellt. Auffällig war, dass kein signifikanter Einfluss bzgl. des biologischen Geschlechts, des Gradings, der N Kategorie oder des Alters ermittelt werden konnte.
In Bezug auf das Gesamtüberleben der Patienten ergaben sich folgende signifikante Risikofaktoren: eine T-Kategorie ≥ T3 (Hazard-Ratio 4,091), ein hohes UICC-Stadium (Hazard Ratio 2,89 für Stadium IIIC), das fehlende initiale therapeutische Ansprechen der Radiochemotherapie (Hazard-Ratio 9,59), ein Karnofsky-Performance-Status ≤ 80 % (Hazard-Ratio 12,23). Ein protektiver Faktor stellte ein längeres tumorfreies Überleben (Hazard-Ratio 0,935) dar.
Die Auswertung des gesamten und des tumorfreien Überlebens hinsichtlich des Befallsmusters der Rezidive ergab, dass mit zusätzlich zum lokalen Rezidiv nachgewiesenem lokoregionären Rezidiv und Fernmetastasen sich sowohl das tumorfreie Überleben (Ein-Jahres-tumorfreies Überleben 52,9 ± 12,1 % vs. 15,0 ± 8,0 %) als auch das Gesamtüberleben (5-Jahres-Gesamtüberleben 75,0 ± 12,5 % vs. 0,0 %) signifikant verringerten.
Im Vergleich der Merkmale der Patienten mit und ohne nachgewiesenen Fernmetastasen ergab sich ein signifikanter Unterschied in Bezug auf den Anteil der Tumore ≥ T3 (75 %).
In der Untergruppe, die mittels abdominoperinealer Rektumexstirpation therapiert wurde, konnte kein signifikanter Unterschied in Bezug auf die rpT-Kategorie, R-Klassikfikation, Pn-, V- und L-Klassifikation festgestellt werden.
Background: This study investigates (1) whether alterations in magnetic resonance imaging (MRI)-based structural global network organization is impaired in patients with major depressive disorder (MDD), (2) whether in-patient treatment including pharmacological, psychological and neurostimulation interventions is linked to changes in structural brain connectivity and (3) whether brain structural changes relate to changes in depression symptomatology.
Methods: One hundred seventy-eight subjects – 109 subjects diagnosed with current MDD and 55 healthy controls (HC) - participated in the present study (baseline + 6-weeks follow up). Fifty-six depressed patients were treated with electroconvulsive therapy (ECT) and 67 received in-patient treatment without ECT. Here, grey matter T1-weighted MRI was used to define nodes and DWI-based tractography to define the connections – or edges – between the nodes creating a structural connectome. Changes over time in depressions symptom severity was measured with the Hamilton Depression Ratings Scale.
Results: MDD patients showed reduced connectivity strength at baseline compared to healthy controls. MDD patients showed a significant increase of connectivity strength over time, an effect that was not detected in HC. An increase of connectivity strength was associated with a decrease in depression symptom severity. These effects were independent of treatment choice, suggesting a nonspecific effect that cannot be traced back to ECT.
Conclusion: We demonstrate an alleviation of structural brain dysconnectivity in MDD patients after successful antidepressive treatment, which is most prominent in those patients that show the greatest reduction in depressive symptomatology. This pattern of results suggests neuroplastic mechanisms involved in the successful treatment of depression and should be investigated as a potential treatment target in future studies.
Research Category and Technology and Methods: Clinical Research: 2. Electroconvulsive Therapy (ECT)
Introduction: The COVID-19 pandemic has necessitated a reduction in face-to-face consultations, resulting in significant limitations in healthcare for individuals with depression. To ensure safe and adequate care, e-health services, such as telemedicine, gained a more prominent role. Governments have eased restrictions on the use of telemedicine, enabling healthcare professionals to increasingly offer video and telephone consultations.
Objective: This study examines, 1) possible changes over the course of the pandemic in reported use of video and telephone consultations and intended future use of video consultations with healthcare professionals among adults with diagnosed depression; 2) their attitudes towards video and telephone consultations and perceived barriers towards using e-health after prolonged time of the pandemic; and 3) differences in results between subgroups based on sociodemographic and clinical characteristics.
Methods: Three population-representative online surveys were conducted in Germany at different timepoints (t) during the COVID-19 pandemic. Respondents aged 18–69 years with a professionally diagnosed depression were included in the present analyses (t1: June/July 2020 with n = 1094; t2: February 2021 with n = 1038; t3: September 2021 with n = 1255).
Results: The overall proportion of adults with depression who used video or telephone consultations did not change significantly in the time surveyed (t1: 16.51 %, n = 179; t2: 20.23 %, n = 210; t3: 18.47 %, n = 230). However, among users, reported use of video consultations with a psychotherapist increased significantly from t1 (34.83 %, n = 62) to t3 (44.98 %, n = 102, p = .023). Intended future use of VC for healthcare varied depending on the purpose of the consultation. Significant differences over time were only found for the purpose of using VC to discuss clinical findings, laboratory results and diagnostic analyses with a doctor, with higher intentions reported at t2 during lockdown in Germany. At t3, the majority of adults with depression felt that video and telephone consultations were too impersonal and considered them more as a helpful support rather than an alternative to face-to-face psychotherapy. Key barriers to using e-health were found within the societal context and the lacking support from significant others for using e-health, while knowledge and skills represented facilitators for using e-health.
Conclusion: Despite ambivalent attitudes towards video and telephone consultations among adults with depression, reported use of video consultations with a psychotherapist increased during the COVID-19 pandemic.
We recently described that monoacylglycerol lipase (MGL) is present in the tumor microenvironment (TME), increasing tumor growth. In this study we compare the implications of MGL deficiency in the TME in different tumor types.
We show that subcutaneous injection of KP (KrasLSL-G12D/p53fl/fl, mouse lung adenocarcinoma) or B16-F10 cells (mouse melanoma) induced tumor growth in MGL wild type (WT) and knockout (KO) mice. MGL deficiency in the TME attenuated the growth of KP cell tumors whereas tumors from B16-F10 cells increased in size. Opposite immune cell profiles were detected between the two tumor types in MGL KO mice. In line with their anti-tumorigenic function, the number of CD8+ effector T cells and eosinophils increased in KP cell tumors of MGL KO vs. WT mice whereas their presence was reduced in B16-F10 cell tumors of MGL KO mice. Differences were seen in lipid profiles between the investigated tumor types. 2-arachidonoylglycerol (2-AG) content significantly increased in KP, but not B16-F10 cell tumors of MGL KO vs. WT mice while other endocannabinoid-related lipids remained unchanged. However, profiles of phospho- and lysophospholipids, sphingomyelins and fatty acids in KP cell tumors were clearly distinct to those measured in B16-F10 cell tumors.
Our data indicate that TME-localized MGL impacts tumor growth, as well as levels of 2-AG and other lipids in a tumor specific manner.
Background: Hepatectomy is a complex procedure with high morbidity and mortality.
Early prediction/prevention of major complications is highly valuable for patient care.
Surgical APGAR score (SAS) has been validated to predict post-surgical complications (PCs).
Goal: We aimed to define a simple complications classification following hepatectomy based on a therapy-oriented severity Clavien-Dindo classification (CDC).
Methods: 119 patients undergoing liver resection were included. PCs were determined at follow-up based on CDC. Clinicopathological factors were used to calculate SAS. A receiver-operator characteristic (ROC) curve analysis estimated the predictive value of SAS for PCs. Circulating markers levels of liver injury were analyzed as critical elements on PCs.
Results: SAS (P=0.008), estimated blood-loss (P=0.018) and operation time (P=0.0008) were associated with PCs. SAS was reduced in patients with (+) compared to those without (-) complications (6.64±1.84 vs 5.70±1.79, P=0.0079). The area-under-the-curve was 0.646 by ROC, indicating an acceptable discrimination with 65% possibility to distinguish (-) and (+) groups (P=0.004).
Best cutoff value for SAS was ≤6/≥7, at which sensitivity and specificity were maximal.
ALT/ASL levels were significantly different within the group with 9-10 SAS points (P=0.01 and 0.02).
Conclusion: SAS provides accurate risk stratification for major PCs after hepatectomy, and might help improving the overall patient outcome.
Lungensonographie bei Patienten mit HIV und AIDS
Ziele und Methoden: Vor dem Hintergrund mehrerer retrospektiven Studien und Fallserien zur Anwendung des Lungenultraschalls zur Diagnose HIV-assoziierter Lungenerkrankungen führten wir eine prospektive Studie in der Abteilung für Infektiologie am Klinikum der Goethe-Universität Frankfurt durch. Wir schlossen hierbei Patientinnen und Patienten ein, bei denen durch konventionelle Diagnostik eine Lungenerkrankung nachzuweisen oder auszuschließen war, und führten zeitnah zur konventionellen Diagnostik verblindete Ultraschalluntersuchungen durch. Diese wurden zudem verblindet von zwei weiteren Ultraschalluntersuchern reevaluiert, basierend darauf wurde die Interrater-Reliabilität zwischen den drei Befunden errechnet. Die konventionell-radiologischen Untersuchungen wurden verblindet von einem weiteren Radiologen befundet.
Ergebnisse: Wir untersuchten 80 HIV-positive Patienten, von denen 54 nachweislich eine pulmonale Erkrankung hatten. Die häufigsten Diagnosen waren Pneumocystis jirovecii-Pneumonien (21 Fälle), bakterielle Pneumonien (17 Fälle) und andere Diagnosen (16 Fälle). Die Lungenultraschalluntersuchungen zeigten bei 90.7% der Patienten mit pulmonaler Diagnose und bei 34.5% der lungengesunden Patienten Auffälligkeiten. Die CT-Untersuchungen fanden Pathologien in 97.5% der Erkrankten und 27.3% der Gesunden. Röntgenuntersuchungen zeigten bei 78.1% der Erkrankten und bei 25% der Gesunden pathologische Befunde. Die häufigsten Pathologien in allen Modalitäten waren interstitielle Veränderungen. Diese zeigten in der Lungenultraschalluntersuchung keinen signifikanten Unterschied zwischen den verschiedenen Lungenerkrankungen, waren jedoch signifikant häufiger als bei Patienten ohne Lungenerkrankung. Konsolidierungen und Pleuraergüsse waren im Lungenultraschall zwar häufiger bei Erkrankten, aber nicht signifikant gegenüber den Gesunden. Die Interrater-Reliabilität des Lungenultraschalls war hoch für interstitielle Pathologien (ICC=0.82) ohne nennenswerte Änderungen im Studienverlauf (r=-0.11), und niedriger bei Konsolidierungen und Ergüssen (jeweils κ=0.12) mit positivem Zusammenhang zur Studiendauer (r=0.88 für Konsolidierungen, r=0.37 für Ergüsse).
Diskussion: Lungenerkrankungen sind bei HIV-Patienten im Ultraschall am häufigsten durch interstitielle Pathologien nachzuweisen, diese allein erlauben allerdings keine Unterscheidung zwischen verschiedenen Erkrankungen. Die Sensitivität der Ultraschalluntersuchung ist geringer als die der Computertomographie, jedoch höher als die Sensitivität des Röntgens. Interstitielle Pathologien werden bereits nach kurzer Lernphase reliabel identifiziert. Konsolidierungen und Ergüsse waren seltenere Befunde, und scheinen eine längere Lernphase zu benötigen, da die Interrater-Reliabilität im Verlauf der Studie ansteigt. In allen Modalitäten zeigten auch gesunde Patienten bildgebend Auffälligkeiten, was den positiv prädiktiven Wert bei niedrigerer Prävalenz negativ beeinflussen kann.
Lungensonographie bei Patienten mit COVID-19
Ziele und Methoden: Im Rahmen der seit Ende 2019 andauernden COVID-19 Pandemie boten sich die erlernten Sonographiekenntnisse für die bettseitige Diagnostik dieser neuen Lungenerkrankung an. Aufgrund der Pathogenität des neuartigen Erregers wandten wir die Lungensonographie zur Diagnostik unter Isolationsbedingungen an, um Transporte für konventionelle Bildgebung zu reduzieren. Bettseitig durchgeführte Lungensonographien bei SARS-CoV-2-positiven Patientinnen und Patienten wurden mit durchgeführter konventioneller Bildgebung im Rahmen dieser retrospektiven Fallserie verglichen.
Ergebnisse: Die Ultraschalluntersuchungen von 17 Patientinnen und Patienten wurden ausgewertet, hiervon zeigten 14 Untersuchungen Pathologien. In neun Fällen wurden interstitielle Pathologien beobachtet, fünf dieser Fälle zeigten zusätzlich Konsolidierungen.
Drei Patienten hatten außer Konsolidierungen keine weiteren Pathologien. Pleuraergüsse wurden in drei Fällen beobachtet, Pleuraplaques bei einem Patienten. In 5 von 7 Fällen mit auffälligem Thorax-CT zeigte die Lungensonographie Pathologien. Die Lungensonographie zeigte in jedem Fall Pathologien, in dem auch das Röntgen auffällig war.
Diskussion: Diese Fallserie zeigt die Anwendung der Sonographie als bettseitige Lungendiagnostik unter Isolationsbedingungen. Interstitielle Pathologien und Konsolidierungen scheinen die relevantesten Hinweise auf eine pulmonale Beteiligung im Rahmen der COVID-19-Erkrankung zu sein. Wegen der geringen Fallzahl und des retrospektiven Designs ist die Aussagekraft dieser Substudie gering, jedoch zeigt sie die Umsetzbarkeit der Sonographie als bildgebendes Verfahren, wenn aufgrund notwendiger Isolationsmaßnahmen und hoher Fallzahlen konventionelle Bildgebung erschwert ist.