Publications by authors named "Michael Schmitt"

439 Publications

Dual Effects of Cyclooxygenase Inhibitors in Combination With CD19.CAR-T Cell Immunotherapy.

Front Immunol 2021 26;12:670088. Epub 2021 May 26.

Department of Internal Medicine V, University Clinic Heidelberg, Heidelberg University, Heidelberg, Germany.

Chimeric antigen receptor T (CAR-T) cells targeting CD19 came into clinical practice for the treatment of B cell lymphoma in 2018. However, patients being treated for B cell lymphoma often suffer from comorbidities such as chronic pain, cardiovascular diseases and arthritis. Thus, these patients frequently receive concomitant medications that include nonsteroidal anti-inflammatory drugs (NSAIDs) like cyclooxygenase (COX) inhibitors. Celecoxib, a selective COX-2 inhibitor, and aspirin, a non-selective COX-1 and COX-2 inhibitor, are being used as anti-inflammatory, analgesic and anti-pyretic drugs. In addition, several studies have also focused on the anti-neoplastic properties of COX-inhibitors. As the influence of COX-inhibitors on CD19.CAR-T cells is still unknown, we investigated the effect of celecoxib and aspirin on the quantity and quality of CD19.CAR-T cells at different concentrations with special regard to cytotoxicity, activation, cytokine release, proliferation and exhaustion. A significant effect on CAR-T cells could be observed for 0.1 mmol/L of celecoxib and for 4 mmol/L of aspirin. At these concentrations, we found that both COX-inhibitors could induce intrinsic apoptosis of CD19.CAR-T cells showing a significant reduction in the ratio of JC-10 red to JC-10 green CAR-T cells from 6.46 ± 7.03 (mean ± SD) to 1.76 ± 0.67 by celecoxib and to 4.41 ± 0.32 by aspirin, respectively. Additionally, the ratios of JC-10 red to JC-10 green Daudi cells were also decreased from 3.41 ± 0.30 to 0.77 ± 0.06 by celecoxib and to 1.26 ± 0.04 by aspirin, respectively. Although the cytokine release by CD19.CAR-T cells upon activation was not hampered by both COX-inhibitors, activation and proliferation of CAR-T cells were significantly inhibited diminishing the NF-ĸB signaling pathway by a significant down-regulation of expression of CD27 on CD4 and CD8 CAR-T cells, followed by a clear decrease of phosphorylated NF-ĸB p65 in both CD4 and CD8 CAR-T cells by a factor of 1.8. Of note, COX-inhibitors hampered expansion and induced exhaustion of CAR-T cells in an antigen stress assay. Collectively, our findings indicate that the use of COX-inhibitors is a double-edged sword that not only induces apoptosis in tumor cells but also impairs the quantity and quality of CAR-T cells. Therefore, COX-inhibitors should be used with caution in patients with B cell lymphoma under CAR-T cell therapy.
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http://dx.doi.org/10.3389/fimmu.2021.670088DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8189155PMC
May 2021

Diagnosing Systemic Amyloidosis Presenting as Carpal Tunnel Syndrome: A Risk Nomogram to Guide Biopsy at Time of Carpal Tunnel Release.

J Bone Joint Surg Am 2021 Jun 7. Epub 2021 Jun 7.

Division of Plastic and Reconstructive Surgery, Department of Surgery, Oregon Health & Science University, Portland, Oregon.

Background: As carpal tunnel syndrome often precedes other signs of systemic amyloidosis, tenosynovial biopsy at the time of carpal tunnel release may facilitate early diagnosis and treatment. However, evidence-based guidelines for amyloidosis screening during carpal tunnel release have not been established. We sought to develop a predictive model for amyloidosis after carpal tunnel release to inform screening efforts.

Methods: We performed a retrospective cohort study of adults without known amyloidosis undergoing at least 1 carpal tunnel release from 2000 to 2019 with use of the national Veterans Health Administration database. After estimating the cumulative incidence of amyloidosis after carpal tunnel release, we identified risk factors, constructed a predictive nomogram based on a multivariable subdistribution-hazard competing-risks model, and performed cross-validation.

Results: Among 89,981 patients undergoing at least 1 carpal tunnel release, 310 were subsequently diagnosed with amyloidosis at a median interval of 4.5 years, corresponding to a cumulative incidence of 0.55% (95% confidence interval [CI]: 0.47% to 0.63%) at 10 years. Amyloidosis diagnosis following carpal tunnel release was associated with an increased hazard of heart failure (hazard ratio [HR], 4.68; 95% CI: 4.26 to 5.55) and death (HR, 1.27; 95% CI: 1.07 to 1.51) after adjustment for potential confounders. Age, male sex, Black race, monoclonal gammopathy of undetermined significance or multiple myeloma, rheumatoid arthritis, atrial fibrillation, spinal stenosis, and bilateral carpal tunnel syndrome were independently associated with increased risk of amyloidosis diagnosis and were included in the risk nomogram.

Conclusions: Amyloidosis diagnosis after carpal tunnel release is rare but is associated with poor outcomes. We present an amyloidosis-risk nomogram to help guide tenosynovial biopsy at time of carpal tunnel release.

Level Of Evidence: Prognostic Level IV. See Instructions for Authors for a complete description of levels of evidence.
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http://dx.doi.org/10.2106/JBJS.20.02093DOI Listing
June 2021

Recent developments in the social identity approach to the psychology of climate change.

Curr Opin Psychol 2021 May 1;42:95-101. Epub 2021 May 1.

Simon Fraser University, Canada.

We review recent literature on the social psychology of climate change, focusing on the application of social identity theory and self-categorization theory. These two theories, together forming the social identity approach, point to ways in which collective identities influence responses to climate change. Recent research demonstrates that collective identities influence attitudes, beliefs and behavior relevant to climate change, and they do this through processes such as group norms and social influence, collective efficacy, and collective emotions. The SIA suggests that, in general, people are motivated to protect the identity and status of their ingroups. Indeed, recent studies find that groups who are of higher status, and thus have more to gain from protecting the status quo, tend to be less concerned about addressing climate change than lower status groups, who are more likely to be harmed by climate change. However, individuals from both high and low status groups will be more likely to work towards pro-environmental social change when they perceive current social systems that perpetuate climate change as illegitimate and when they can imagine cognitive alternatives to the status quo, where humans have a more sustainable relationship with nature.
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http://dx.doi.org/10.1016/j.copsyc.2021.04.009DOI Listing
May 2021

Computational tissue staining of non-linear multimodal imaging using supervised and unsupervised deep learning.

Biomed Opt Express 2021 Apr 23;12(4):2280-2298. Epub 2021 Mar 23.

Institute of Physical Chemistry and Abbe Center of Photonics, Friedrich-Schiller-University, Jena, Germany.

Hematoxylin and Eosin (H&E) staining is the 'gold-standard' method in histopathology. However, standard H&E staining of high-quality tissue sections requires long sample preparation times including sample embedding, which restricts its application for 'real-time' disease diagnosis. Due to this reason, a label-free alternative technique like non-linear multimodal (NLM) imaging, which is the combination of three non-linear optical modalities including coherent anti-Stokes Raman scattering, two-photon excitation fluorescence and second-harmonic generation, is proposed in this work. To correlate the information of the NLM images with H&E images, this work proposes computational staining of NLM images using deep learning models in a supervised and an unsupervised approach. In the supervised and the unsupervised approach, conditional generative adversarial networks (CGANs) and cycle conditional generative adversarial networks (cycle CGANs) are used, respectively. Both CGAN and cycle CGAN models generate pseudo H&E images, which are quantitatively analyzed based on mean squared error, structure similarity index and color shading similarity index. The mean of the three metrics calculated for the computationally generated H&E images indicate significant performance. Thus, utilizing CGAN and cycle CGAN models for computational staining is beneficial for diagnostic applications without performing a laboratory-based staining procedure. To the author's best knowledge, it is the first time that NLM images are computationally stained to H&E images using GANs in an unsupervised manner.
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http://dx.doi.org/10.1364/BOE.415962DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8086483PMC
April 2021

Excited state dipole moments and lifetimes of 2-cyanoindole from rotationally resolved electronic Stark spectroscopy.

Phys Chem Chem Phys 2021 May 19;23(17):10196-10204. Epub 2021 Mar 19.

Heinrich-Heine-Universität, Institut für Physikalische Chemie I, D-40225 Düsseldorf, Germany.

The permanent dipole moments of 2-cyanoindole (cyanoindole = CNI) in its ground and lowest excited singlet states have been determined from rotationally resolved electronic Stark spectroscopy under jet-cooled conditions. From the orientation of the transition dipole moment and the geometry changes upon electronic excitation the lowest excited singlet state could be shown to be of L-symmetry. The general statement, that the L-state has the larger permanent dipole moment of the two lowest excited singlet states, will be challenged in this contribution. On the basis of the different electronic nature of the first excited singlet state the behavior of 2-, 3-, 4- and 5-CNI is discussed. The excited state lifetime of isolated 2-CNI in the gas phase has been determined to be 9.4 ns. This value is compared to the excited state lifetime in ethyl acetate solution of 2.6 ns, which was quantified with a Strickler-Berg analysis. Using water as solvent shortens the 2-CNI lifetime to <40 ps. The reason for this drastic shortening is discussed in detail. Additionally, the rotationally resolved electronic spectrum of 2-CNI(1-d) has been measured and analyzed.
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http://dx.doi.org/10.1039/d1cp00097gDOI Listing
May 2021

l-citrulline: A preclinical safety biomarker for the small intestine in rats and dogs in repeat dose toxicity studies.

J Pharmacol Toxicol Methods 2021 May 1;110:107068. Epub 2021 May 1.

Merck KGaA, Chemical and Preclinical Safety, Darmstadt, Germany.

Introduction: Gastrointestinal (GI) toxicity is still an issue within drug development, especially for novel oncology drugs. The identification of GI mucosal damage at an early stage with high sensitivity and specificity across preclinical species and humans remains difficult. To date, in preclinical studies, no qualified mechanistic, diagnostic or prognostic biomarkers exist for GI mucosal toxicity. l-citrulline is one of the most promising biomarker candidates used in clinical settings to quantify enterocyte integrity in various small intestinal diseases. l-citrulline is an intermediate metabolic amino acid produced mainly by functional enterocytes of the small intestine, whereby enterocyte loss will cause a drop in circulating l-citrulline.

Methods: In several repeat-dose toxicity studies, plasma l-citrulline has been evaluated as a potential safety biomarker for intestinal toxicity in beagle dogs and Wistar (Han) rats treated with different oncological drug candidates in drug development. Clinical observations and body weight determinations were performed during the pretreatment, treatment and treatment-free recovery period as well as toxicokinetic, gross and histopathology examinations. The quantitative determination of plasma l-citrulline levels during the pretreatment (only dogs), treatment and treatment-free recovery period were performed using an HPLC MS/MS assay. In cynomolgus monkeys, the first investigations on baseline l-citrulline levels were performed.

Results: In dogs, a dose- and exposure-dependent decrease of up to 50% in plasma l-citrulline was seen without histopathological alterations. However, a decrease of more than 50% in comparison to the individual animal pretreatment value of l-citrulline correlated very well with histopathological findings (intestinal crypt necrosis, villus atrophy, enterocyte loss) and clinical signs (bloody faeces and diarrhoea). During a treatment-free recovery period, a trend of increasing levels was observed in dogs. In rats, absolute l-citrulline plasma levels of treated animals decreased compared to the values of the concurrent control group. This decrease also correlated with the histopathological findings in the small intestine (single cell necrosis and mucosa atrophy). Because of a large physiological variation in l-citrulline plasma levels in dogs and rats, a clear cut-off value for absolute l-citrulline levels predictive of intestinal mucosal toxicity was difficult to establish. However, a > 50% decrease in l-citrulline plasma levels during the treatment period strongly correlated with histopathological findings.

Discussion: Based on the performed analysis, a longitudinal investigation of l-citrulline plasma levels for individual animals in the control and treatment groups is essential and pretreatment values of l-citrulline levels in rodents would be highly informative. Overall, further cross-species comparison (Cynomolgus monkey, mouse) and implementation in clinical trials as exploratory biomarker is essential to foster the hypothesis and to understand completely the clinical relevance of l-citrulline as a small intestine biomarker.
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http://dx.doi.org/10.1016/j.vascn.2021.107068DOI Listing
May 2021

Infection Complications after Lymphodepletion and Dosing of Chimeric Antigen Receptor T (CAR-T) Cell Therapy in Patients with Relapsed/Refractory Acute Lymphoblastic Leukemia or B Cell Non-Hodgkin Lymphoma.

Cancers (Basel) 2021 Apr 2;13(7). Epub 2021 Apr 2.

Department of Internal Medicine V-Hematology, Oncology & Rheumatology, University Hospital Heidelberg, 69120 Heidelberg, Germany.

Chimeric antigen receptor T (CAR-T) cell therapy has proven to be very effective in patients with relapsed/refractory acute lymphoblastic leukemia (ALL) and non-Hodgkin lymphoma (NHL). However, infections-related either due to lymphodepletion or the CAR-T cell therapy itself-can result in severe and potentially life-threatening complications, while side effects such as cytokine release syndrome (CRS) might complicate differential diagnosis. Sixty-seven dosings of CAR-T cells in sixty adult patients with NHL (85%) and ALL (15%) receiving CAR-T cell therapy were assessed for infectious complications. Almost two-thirds of patients (61%) developed fever following lymphodepletion and CAR-T cell dosing. Microbiological or radiological findings were observed in 25% of all cases (bacterial 12%, viral 5%, fungal 8%). Inpatient infections were associated with more lines of therapy and more severe CRS. However, overall serious complications were rare after CAR-T therapy, with one patient dying of infection. Pathogen detection after inpatient stay was infrequent and mostly occurred in the first 90 days after dosing. Infections in CAR-T cell treated patents are common. Fast and suitable identification and treatment are crucial in these heavily pretreated and immunocompromised patients. In most cases infectious complications are manageable. Nonetheless, standardized anti-infective prophylaxis and supportive therapy are mandatory to reduce morbidity and mortality in CAR-T cell therapy.
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http://dx.doi.org/10.3390/cancers13071684DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8038233PMC
April 2021

Kinetic-Model-Free Analysis of Transient Absorption Spectra Enabled by 2D Correlation Analysis.

J Phys Chem Lett 2021 May 23;12(17):4148-4153. Epub 2021 Apr 23.

Leibniz Institute of Photonic Technology, Albert-Einstein-Str. 9, 07745 Jena, Germany.

Here, we present, to the best of our knowledge for the first time, a systematic study of utilizing 2D correlation analysis in the field of femtosecond transient absorption (fs-TA) spectroscopy. We present that the application of 2D correlation spectroscopy (2DCOS) to fs-TA spectroscopy enables a model-free means to analyze excited state kinetics, which is demonstrated on the model system [(tbbpy)Ru(dppz)] in different solvents. We show that TA-2DCOS is able to determine the number of processes contributing to the time-resolved spectral changes in fs-TA data sets, as well as extract the spectral response of these components. Overall, the results show that TA-2DCOS leads to the same results as obtained with methods relying on global lifetime analysis or multivariate curve resolution but without the need to specify a predetermined kinetic model. The work presented therefore highlights the potential of TA-2DCOS as a model-free approach for analyzing fs-TA spectral data sets.
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http://dx.doi.org/10.1021/acs.jpclett.1c00835DOI Listing
May 2021

A vaccine targeting mutant IDH1 in newly diagnosed glioma.

Nature 2021 Apr 24;592(7854):463-468. Epub 2021 Mar 24.

Neurology Clinic, Heidelberg University Hospital, University of Heidelberg, Heidelberg, Germany.

Mutated isocitrate dehydrogenase 1 (IDH1) defines a molecularly distinct subtype of diffuse glioma. The most common IDH1 mutation in gliomas affects codon 132 and encodes IDH1(R132H), which harbours a shared clonal neoepitope that is presented on major histocompatibility complex (MHC) class II. An IDH1(R132H)-specific peptide vaccine (IDH1-vac) induces specific therapeutic T helper cell responses that are effective against IDH1(R132H) tumours in syngeneic MHC-humanized mice. Here we describe a multicentre, single-arm, open-label, first-in-humans phase I trial that we carried out in 33 patients with newly diagnosed World Health Organization grade 3 and 4 IDH1(R132H) astrocytomas (Neurooncology Working Group of the German Cancer Society trial 16 (NOA16), ClinicalTrials.gov identifier NCT02454634). The trial met its primary safety endpoint, with vaccine-related adverse events restricted to grade 1. Vaccine-induced immune responses were observed in 93.3% of patients across multiple MHC alleles. Three-year progression-free and death-free rates were 0.63 and 0.84, respectively. Patients with immune responses showed a two-year progression-free rate of 0.82. Two patients without an immune response showed tumour progression within two years of first diagnosis. A mutation-specificity score that incorporates the duration and level of vaccine-induced IDH1(R132H)-specific T cell responses was associated with intratumoral presentation of the IDH1(R132H) neoantigen in pre-treatment tumour tissue. There was a high frequency of pseudoprogression, which indicates intratumoral inflammatory reactions. Pseudoprogression was associated with increased vaccine-induced peripheral T cell responses. Combined single-cell RNA and T cell receptor sequencing showed that tumour-infiltrating CD40LG and CXCL13 T helper cell clusters in a patient with pseudoprogression were dominated by a single IDH1(R132H)-reactive T cell receptor.
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http://dx.doi.org/10.1038/s41586-021-03363-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8046668PMC
April 2021

In vivo coherent anti-Stokes Raman scattering microscopy reveals vitamin A distribution in the liver.

J Biophotonics 2021 Jun 6;14(6):e202100040. Epub 2021 Apr 6.

Institute of Physical Chemistry and Abbe Center of Photonics, Friedrich Schiller University, Jena, Germany.

Here we present a microscope setup for coherent anti-Stokes Raman scattering (CARS) imaging, devised to specifically address the challenges of in vivo experiments. We exemplify its capabilities by demonstrating how CARS microscopy can be used to identify vitamin A (VA) accumulations in the liver of a living mouse, marking the positions of hepatic stellate cells (HSCs). HSCs are the main source of extracellular matrix protein after hepatic injury and are therefore the main target of novel nanomedical strategies in the development of a treatment for liver fibrosis. Their role in the VA metabolism makes them an ideal target for a CARS-based approach as they store most of the body's VA, a class of compounds sharing a retinyl group as a structural motive, a moiety that is well known for its exceptionally high Raman cross section of the C═C stretching vibration of the conjugated backbone.
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http://dx.doi.org/10.1002/jbio.202100040DOI Listing
June 2021

CD70-specific CAR T-cells have potent activity against Acute Myeloid Leukemia (AML) without HSC toxicity.

Blood 2021 Mar 15. Epub 2021 Mar 15.

Baylor College of Medicine, Houston, Texas, United States.

The prognosis of patients with acute myeloid leukemia (AML) remains dismal highlighting the need for novel innovative treatment strategies. The application of chimeric antigen receptor (CAR) T-cell therapy to AML patients has been limited in particular by the lack of a tumor-specific target antigen. CD70 is a promising antigen to target AML as it is expressed on the majority of leukemic blasts, whereas little or no expression is detectable in normal bone marrow samples. To target CD70 on AML cells, we generated a panel of CD70-CARs that contained a common single chain variable fragment (scFv) for antigen detection but differed in size and flexibility of the extracellular spacer, and in the transmembrane and the co-stimulatory domains. These CD70scFv CARs were compared with a CAR construct that contained the human CD27, the ligand of CD70 fused to the CD3z chain (CD27z). The structural composition of the CAR strongly influenced expression levels, viability, expansion and cytotoxic capacities of CD70scFv CAR T-cells, but the CD27z-CAR T-cells demonstrated superior proliferation and anti-tumor activity in vitro and in vivo, compared to all CD70scFv-CARs. While CD70-CAR T-cells recognized activated virus-specific T-cells (VSTs) that expressed CD70, they did not prevent colony formation by normal hematopoietic stem cells (HSCs). Thus, CD70-targeted immunotherapy is a promising new treatment strategy for patients with CD70-positive AML that does not affect normal hematopoiesis but will require monitoring of virus-specific T-cell responses.
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http://dx.doi.org/10.1182/blood.2020008221DOI Listing
March 2021

An Endoplasmic Reticulum Specific Pro-amplifier of Reactive Oxygen Species in Cancer Cells.

Angew Chem Int Ed Engl 2021 05 8;60(20):11158-11162. Epub 2021 Apr 8.

Friedrich-Alexander University Erlangen-Nürnberg (FAU), Department of Chemistry and Pharmacy, Organic Chemistry Chair II, Nikolaus-Fiebiger-Str. 10, 91058, Erlangen, Germany.

The folding and export of proteins and hydrolysis of unfolded proteins are disbalanced in the endoplasmic reticulum (ER) of cancer cells, leading to so-called ER stress. Agents further augmenting this effect are used as anticancer drugs including clinically approved proteasome inhibitors bortezomib and carfilzomib. However, these drugs can affect normal cells, which also rely strongly on ER functions, leading, for example, to accumulation of reactive oxygen species (ROS). To address this problem, we have developed ER-targeted prodrugs activated only in cancer cells in the presence of elevated ROS amounts. These compounds are conjugates of cholic acid with N-alkylaminoferrocene-based prodrugs. We confirmed their accumulation in the ER of cancer cells, their anticancer efficacy, and cancer cell specificity. These prodrugs induce ER stress, attenuate mitochondrial membrane potential, and generate mitochondrial ROS leading to cell death via necrosis. We also demonstrated that the new prodrugs are activated in vivo in Nemeth-Kellner lymphoma (NK/Ly) murine model.
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http://dx.doi.org/10.1002/anie.202100054DOI Listing
May 2021

Peptide Vaccination against Cytomegalovirus Induces Specific T Cell Response in Responses in CMV Seronegative End-Stage Renal Disease Patients.

Vaccines (Basel) 2021 Feb 6;9(2). Epub 2021 Feb 6.

Department of Internal Medicine V, University of Heidelberg, 69117 Heidelberg, Germany.

Introduction: Cytomegalovirus (CMV) reactivation occurs in seronegative patients after solid organ transplantation (SOT) particularly from seropositive donors and can be lethal. Generation of CMV-specific T cells helps to prevent CMV reactivation. Therefore, we initiated a clinical phase I CMVpp65 peptide vaccination trial for seronegative end-stage renal disease patients waiting for kidney transplantation.

Methods: The highly immunogenic nonamer peptide NLVPMVATV derived from CMV phosphoprotein 65(CMVpp65) in a water-in-oil emulsion (Montanide™) plus imiquimod (Aldara™) as an adjuvant was administered subcutaneously four times biweekly. Clinical course as well as immunological responses were monitored using IFN-γ ELISpot assays and flow cytometry for CMV-specific CD8 T cells.

Results: Peptide vaccination was well tolerated, and no drug-related serious adverse events were detected except for Grade I-II local skin reactions. Five of the 10 patients (50%) mounted any immune response (responders) and 40% of the patients presented CMV-specific CD8 T cell responses elicited by these prophylactic vaccinations. No responders experienced CMV reactivation in the 18 months post-transplantation, while all non-responders reactivated.

Conclusion: CMVpp65 peptide vaccination was safe, well tolerated, and clinically encouraging in seronegative end-stage renal disease patients waiting for kidney transplantation. Further studies with larger patient cohorts are planned.
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http://dx.doi.org/10.3390/vaccines9020133DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7915922PMC
February 2021

Evaluation of Production Protocols for the Generation of NY-ESO-1-Specific T Cells.

Cells 2021 01 14;10(1). Epub 2021 Jan 14.

Department of Internal Medicine V, Heidelberg University Hospital, 69120 Heidelberg, Germany.

NY-ESO-1-specific T cells have shown promising activity in the treatment of soft tissue sarcoma (STS). However, standardized protocols for their generation are limited. Particularly, cost-effectiveness considerations of cell production protocols are of importance for conducting clinical studies. In this study, two different NY-ESO-1-specific T cell production protocols were compared. Major differences between protocols 1 and 2 include culture medium, interleukin-2 and retronectin concentrations, T cell activation strategy, and the transduction process. NY-ESO-1-specific T cells generated according to the two protocols were investigated for differences in cell viability, transduction efficiency, T cell expansion, immunophenotype as well as functionality. NY-ESO-1-specific T cells showed similar viability and transduction efficiency between both protocols. Protocol 1 generated higher absolute numbers of NY-ESO-1-specific T cells. However, there was no difference in absolute numbers of NY-ESO-1-specific T cell subsets with less-differentiated phenotypes accounting for efficient in vivo expansion and engraftment. Furthermore, cells generated according to protocol 1 displayed higher capacity of TNF-α generation, but lower cytotoxic capacities. Overall, both protocols provided functional NY-ESO-1-specific T cells. However, compared to protocol 1, protocol 2 is advantageous in terms of cost-effectiveness. Cell production protocols should be designed diligently to achieve a cost-effective cellular product for further clinical evaluation.
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http://dx.doi.org/10.3390/cells10010152DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7828728PMC
January 2021

Intra-host changes in Kaposi sarcoma-associated herpesvirus genomes in Ugandan adults with Kaposi sarcoma.

PLoS Pathog 2021 01 19;17(1):e1008594. Epub 2021 Jan 19.

University of Washington, Department of Microbiology, Seattle, Washington, United States of America.

Intra-host tumor virus variants may influence the pathogenesis and treatment responses of some virally-associated cancers. However, the intra-host variability of Kaposi sarcoma-associated herpesvirus (KSHV), the etiologic agent of Kaposi sarcoma (KS), has to date been explored with sequencing technologies that possibly introduce more errors than that which occurs in the viral population, and these studies have only studied variable regions. Here, full-length KSHV genomes in tumors and/or oral swabs from 9 Ugandan adults with HIV-associated KS were characterized. Furthermore, we used deep, short-read sequencing using duplex unique molecular identifiers (dUMI)-random double-stranded oligonucleotides that barcode individual DNA molecules before library amplification. This allowed suppression of PCR and sequencing errors to ~10-9/base as well as afforded accurate determination of KSHV genome numbers sequenced in each sample. KSHV genomes were assembled de novo, and rearrangements observed were confirmed by PCR and Sanger sequencing. 131-kb KSHV genome sequences, excluding major repeat regions, were successfully obtained from 23 clinical specimens, averaging 2.3x104 reads/base. Strikingly, KSHV genomes were virtually identical within individuals at the point mutational level. The intra-host heterogeneity that was observed was confined to tumor-associated KSHV mutations and genome rearrangements, all impacting protein-coding sequences. Although it is unclear whether these changes were important to tumorigenesis or occurred as a result of genomic instability in tumors, similar changes were observed across individuals. These included inactivation of the K8.1 gene in tumors of 3 individuals and retention of a region around the first major internal repeat (IR1) in all instances of genomic deletions and rearrangements. Notably, the same breakpoint junctions were found in distinct tumors within single individuals, suggesting metastatic spread of rearranged KSHV genomes. These findings define KSHV intra-host heterogeneity in vivo with greater precision than has been possible in the past and suggest the possibility that aberrant KSHV genomes may contribute to aspects of KS tumorigenesis. Furthermore, study of KSHV with use of dUMI provides a proof of concept for utilizing this technique for detailed study of other virus populations in vivo.
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http://dx.doi.org/10.1371/journal.ppat.1008594DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7845968PMC
January 2021

Shape-Memory Metallopolymers Based on Two Orthogonal Metal-Ligand Interactions.

Adv Mater 2021 Feb 14;33(7):e2006655. Epub 2021 Jan 14.

Laboratory of Organic and Macromolecular Chemistry (IOMC), Friedrich Schiller University Jena, Humboldstr. 10, Jena, 07743, Germany.

A new shape-memory polymer is presented, in which both the stable phase as well as the switching unit consist of two different metal complexes. Suitable metal ions, which simultaneously form labile complexes with histidine and stable ones with terpyridine ligands, are identified via isothermal titration calorimetry (ITC) measurements. Different copolymers are synthesized, which contain butyl methacrylate as the main monomer and the metal-binding ligands in the side chains. Zn(TFMS) and NiCl are utilized for the dual crosslinking, resulting in the formation of metallopolymer networks. The switching temperature can simply be tuned by changing the composition as well as by the choice of the metal ion. Strain fixity rates (about 99%) and very high strain recovery rates (up to 95%) are achieved and the mechanism is revealed using different techniques such as Raman spectroscopy.
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http://dx.doi.org/10.1002/adma.202006655DOI Listing
February 2021

FLIM data analysis based on Laguerre polynomial decomposition and machine-learning.

J Biomed Opt 2021 Jan;26(2)

University of Jena, Institute of Physical Chemistry, Abbe Center of Photonics, Jena, Germany.

Significance: The potential of fluorescence lifetime imaging microscopy (FLIM) is recently being recognized, especially in biological studies. However, FLIM does not directly measure the lifetimes, rather it records the fluorescence decay traces. The lifetimes and/or abundances have to be estimated from these traces during the phase of data processing. To precisely estimate these parameters is challenging and requires a well-designed computer program. Conventionally employed methods, which are based on curve fitting, are computationally expensive and limited in performance especially for highly noisy FLIM data. The graphical analysis, while free of fit, requires calibration samples for a quantitative analysis.

Aim: We propose to extract the lifetimes and abundances directly from the decay traces through machine learning (ML).

Approach: The ML-based approach was verified with simulated testing data in which the lifetimes and abundances were known exactly. Thereafter, we compared its performance with the commercial software SPCImage based on datasets measured from biological samples on a time-correlated single photon counting system. We reconstructed the decay traces using the lifetime and abundance values estimated by ML and SPCImage methods and utilized the root-mean-squared-error (RMSE) as marker.

Results: The RMSE, which represents the difference between the reconstructed and measured decay traces, was observed to be lower for ML than for SPCImage. In addition, we could demonstrate with a three-component analysis the high potential and flexibility of the ML method to deal with more than two lifetime components.

Conclusions: The ML-based approach shows great performance in FLIM data analysis.
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http://dx.doi.org/10.1117/1.JBO.26.2.022909DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7790506PMC
January 2021

Novel Biobased Self-Healing Ionomers Derived from Itaconic Acid Derivates.

Macromol Rapid Commun 2021 Apr 28;42(8):e2000636. Epub 2020 Dec 28.

Laboratory of Organic and Macromolecular Chemistry (IOMC), Friedrich Schiller University Jena, Humboldstr. 10, Jena, 07743, Germany.

This article presents novel biobased ionomers featuring self-healing abilities. These smart materials are synthesized from itaconic acid derivates. Large quantities of itaconic acid can be produced from diverse biomass like corn, rice, and others. This study presents a comprehensive investigation of their thermal and mechanical properties via differential scanning calorimetry (DSC), thermo gravimetric analysis (TGA), and FT-Raman and FT-IR measurements as well as dynamic mechanic analysis. Within all these measurements, different kinds of structure-property relationships could be derived from these measurements. For example, the proportion of ionic groups enormously influences the self-healing efficiency. The investigation of the self-healing abilities reveals healing efficiencies up to 99% in 2 h at 90 °C for the itaconic acid based ionomer with the lowest ionic content.
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http://dx.doi.org/10.1002/marc.202000636DOI Listing
April 2021

Pre-sensitization of Malignant B Cells Through Venetoclax Significantly Improves the Cytotoxic Efficacy of CD19.CAR-T Cells.

Front Immunol 2020 9;11:608167. Epub 2020 Dec 9.

Department of Internal Medicine V, University Clinic Heidelberg, Heidelberg University, Heidelberg, Germany.

Chimeric antigen receptor (CAR) T cell therapy has shown promising responses in patients with refractory or relapsed aggressive B-cell malignancies that are resistant to conventional chemotherapy or stem cell transplantation. A potentially combinatorial therapeutic strategy may be the inhibition of anti-apoptotic Bcl-2 family proteins, overexpressed in most cancer cells. In this study we investigated the combination of 3rd-generation CD19.CAR-T cells and the BH3 mimetics venetoclax, a Bcl-2 inhibitor, or S63845, a Mcl-1 inhibitor, under three different treatment conditions: pre-sensitization of cancer cells with BH3 mimetics followed by CAR-T cell treatment, simultaneous combination therapy, and the administration of BH3 mimetics after CAR-T cell treatment. Our results showed that administration of CAR-T cells and BH3 mimetics had a significant effect on the quantity and quality of CD19.CAR-T cells. The administration of BH3 mimetics prior to CAR-T cell therapy exerted an enhanced cytotoxic efficacy by upregulating the CD19 expression and pro-apoptotic proteins in highly sensitive tumor cells, and thereby improving both CD19.CAR-T cell cytotoxicity and persistence. In simultaneous and post-treatment approaches, however, the quantity of CAR-T cells was adversely affected. Our findings indicate pre-sensitization of highly sensitive tumor cells with BH3 mimetics could enhance the cytotoxic efficacy of CAR-T cell treatment.
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http://dx.doi.org/10.3389/fimmu.2020.608167DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7756123PMC
December 2020

Spatially Resolving the Enhancement Effect in Surface-Enhanced Coherent Anti-Stokes Raman Scattering by Plasmonic Doppler Gratings.

ACS Nano 2021 Jan 23;15(1):809-818. Epub 2020 Dec 23.

Leibniz Institute of Photonic Technology, Albert-Einstein Strasse 9, 07745 Jena, Germany.

Well-designed plasmonic nanostructures can mediate far and near optical fields and thereby enhance light-matter interactions. To obtain the best overall enhancement, structural parameters need to be carefully tuned to obtain the largest enhancement at the input and output frequencies. This is, however, challenging for nonlinear light-matter interactions involving multiple frequencies because obtaining the full picture of structure-dependent enhancement at individual frequencies is not easy. In this work, we introduce the platform of plasmonic Doppler grating (PDG) to experimentally investigate the enhancement effect of plasmonic gratings in the input and output beams of nonlinear surface-enhanced coherent anti-Stokes Raman scattering (SECARS). PDGs are designable azimuthally chirped gratings that provide broadband and spatially dispersed plasmonic enhancement. Therefore, they offer the opportunity to observe and compare the overall enhancement from different combinations of enhancement in individual input and output beams. We first confirm PDG's capability of spatially separating the input and output enhancement in linear surface-enhanced fluorescence and Raman scattering. We then investigate spatially resolved enhancement in nonlinear SECARS, where coherent interaction of the pump, Stokes, and anti-Stokes beams is enhanced by the plasmonic gratings. By mapping the SECARS signal and analyzing the azimuthal angle-dependent intensity, we characterize the enhancement at individual frequencies. Together with theoretical analysis, we show that while simultaneous enhancement in the input and output beams is important for SECARS, the enhancement in the pump and anti-Stokes beams plays a more critical role in the overall enhancement than that in the Stokes beam. This work provides an insight into the enhancement mechanism of plasmon-enhanced spectroscopy, which is important for the design and optimization of plasmonic gratings. The PDG platform may also be applied to study enhancement mechanisms in other nonlinear light-matter interactions or the impact of plasmonic gratings on the fluorescence lifetime.
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http://dx.doi.org/10.1021/acsnano.0c07198DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7944573PMC
January 2021

Diversity of beetles (Coleoptera) in natural and planted saxaul forests () in the South Gobi Desert, Mongolia.

Zookeys 2020 3;1000:59-70. Epub 2020 Dec 3.

General and Systematic Zoology, Zoological Institute and Museum, University of Greifswald, Greifswald, Germany University of Greifswald Greifswald Germany.

We investigated species composition and diversity parameters of beetle communities in two planted saxaul (, black saxaul) forests in Southern Mongolia. We also studied nearby natural areas for comparison. Beetles were mainly collected by pitfall traps. 1064 individuals of 38 species of 22 genera in 4 beetle families were identified from planted plots. In comparison, a total of 1395 beetles belonging to 40 species of 24 genera in seven families were collected and identified from the natural saxaul plots. The most diverse beetle families were darkling beetles (Tenebrionidae, 18 species) and snout beetles (Curculionidae, 15 species) in planted and natural saxaul plots. We recorded several species (, , and ) which are associated with the saxaul tree. A darkling beetle, , was the dominant species in both natural and planted plots of the Nariin Zag forest. There were significant differences in the species richness and abundance between the planted and natural plots of the Ukhaa Zag forest. It is possible that the age of the plantation drove the differences. The higher values of diversity indices and species richness in the planted plots can be explained by the presence of rare species, represented by only one or two individuals. The planted plots and corresponding natural plots within each forest were more similar to each other in species composition and abundance than between forests.
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http://dx.doi.org/10.3897/zookeys.1000.56856DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7728731PMC
December 2020

Perceptual harmony in judgments of group prototypicality and intragroup respect.

PLoS One 2020 22;15(12):e0243821. Epub 2020 Dec 22.

Department of Psychology, University of Kansas, Lawrence, Kansas, United States of America.

We test common sense psychology of intragroup relations whereby people assume that intragroup respect and ingroup prototypicality are positively related. In Study 1a, participants rated a group member as more prototypical if they learned that group member was highly respected rather than disrespected. In Study 1b, participants rated a group member as more respected by other group members if they learned that group member was prototypical rather than unprototypical. As a commonsense psychology of groups, we reasoned that the perceived relationship between prototypicality and intragroup respect would be stronger for cohesive groups compared to incohesive groups. The effect of intragroup respect on perceptions of prototypicality (Study 2a & 2c) and the effect of prototypicality on perceptions of intragroup respect (Study 2b) were generally stronger for participants considering cohesive groups relative to incohesive groups. However, the interaction effect of prototypicality and group cohesion on intragroup respect did fail to replicate in Study 2d. In Studies 3, 4a, and 4b we manipulated the relationship between prototypicality and intragroup respect and found that when these variables were in perceptual harmony participants perceived groups as more cohesive. The results of eight out of nine studies conducted are consistent with the prediction that people make inferences about intragroup respect, prototypicality, and group cohesion in a manner that maintains perceptual harmony.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0243821PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7755273PMC
February 2021

CAR T cells or allogeneic transplantation as standard of care for advanced large B-cell lymphoma: an intent-to-treat comparison.

Blood Adv 2020 12;4(24):6157-6168

Department of Medicine V, University of Heidelberg, Heidelberg, Germany; and.

CD19-directed chimeric antigen receptor (CAR) T-cell treatment has evolved as standard of care (SOC) for multiply relapsed/refractory (R/R) large B-cell lymphoma (LBCL). However, its potential benefit over allogeneic hematopoietic cell transplantation (alloHCT) remains unclear. We compared outcomes with both types of cellular immunotherapy (CI) by intention to treat (ITT). Eligble were all patients with R/R LBCL and institutional tumor board decision recommending SOC CAR T-cell treatment between July 2018 and February 2020, or alloHCT between January 2004 and February 2020. Primary end point was overall survival (OS) from indication. Altogether, 41 and 60 patients for whom CAR T cells and alloHCT were intended, respectively, were included. In both cohorts, virtually all patients had active disease at indication. CI was recommended as part of second-line therapy for 21 alloHCT patients but no CAR T-cell patients. Median OS from indication was 475 days with CAR T cells vs 285 days with alloHCT (P = .88) and 222 days for 39 patients for whom alloHCT beyond second line was recommended (P = .08). Of CAR T-cell and alloHCT patients, 73% and 65%, respectively, proceeded to CI. After CI, 12-month estimates for nonrelapse mortality, relapse incidence, progression-free survival, and OS for CAR T cells vs alloHCT were 3% vs 21% (P = .04), 59% vs 44% (P = .12), 39% vs 33% (P = .97), and 68% vs 54% (P = .32), respectively. In conclusion, CAR T-cell outcomes were not inferior to alloHCT outcomes, whether measured by ITT or from CI administration, supporting strategies preferring CAR T cells over alloHCT as first CI for multiply R/R LBCL.
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http://dx.doi.org/10.1182/bloodadvances.2020003036DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7756983PMC
December 2020

Letermovir prophylaxis is effective in preventing cytomegalovirus reactivation after allogeneic hematopoietic cell transplantation: single-center real-world data.

Ann Hematol 2020 Dec 3. Epub 2020 Dec 3.

Department of Internal Medicine V (Hematology/Oncology/Rheumatology), Heidelberg University Hospital, Im Neuenheimer Feld 410, 69120, Heidelberg, Germany.

Morbidity and mortality after allogeneic hematopoietic cell transplantation (alloHCT) are still essentially affected by reactivation of cytomegalovirus (CMV). We evaluated 80 seropositive patients transplanted consecutively between March 2018 and March 2019 who received letermovir (LET) prophylaxis from engraftment until day +100 and retrospectively compared them with 80 patients without LET allografted between January 2017 and March 2018. The primary endpoint of this study was the cumulative incidence (CI) of clinically significant CMV infection (CS-CMVi) defined as CMV reactivation demanding preemptive treatment or CMV disease. With 14% CI of CS-CMVi at day +100 (11 events) was significantly lower in the LET cohort when compared to the control group (33 events, 41%; HR 0.29; p < 0.001). Whereas therapy with foscarnet could be completely avoided in the LET group, 7 out of 80 patients in the control cohort received foscarnet, resulting in 151 extra in-patient days for foscarnet administration (p = 0.002). One-year overall survival was 72% in the control arm vs 84% in the LET arm (HR 0.75 [95%CI 0.43-1.30]; p < 0.306). This study confirms efficacy and safety of LET for prophylaxis of CS-CMVi after alloHCT in a real-world setting, resulting in a significant patient benefit by reducing hospitalization needs and exposure to potentially toxic antiviral drugs for treatment of CMV reactivation.
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http://dx.doi.org/10.1007/s00277-020-04362-2DOI Listing
December 2020

Determination of excited state dipole moments in solution via thermochromic methods.

MethodsX 2020 18;7:101101. Epub 2020 Oct 18.

Heinrich-Heine-Universität, Institut für Physikalische Chemie I, D-40225 Düsseldorf, Germany.

The method basically combines the existing ideas of excited state dipole moment determination via thermochromic fluorescence spectroscopy with the determination of the solvent cavity volume via concentration dependent density measurements of the solution densities at different weight fractions. Additionally, the determination of the cavity volume in dependence of the solvent temperature is included here, which provides a better accuracy of the excited state dipole moment determination. With this step two major sources of errors are eliminated: the use of the very imprecise Onsager radius and the assumption, that the cavity size is temperature independent.•Thermochromic absorption and fluorescence spectroscopy.•Cavity volume determination by density measurements.•Temperature dependent cavity volume determination.
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http://dx.doi.org/10.1016/j.mex.2020.101101DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7649500PMC
October 2020

A Randomized Open label Phase-II Clinical Trial with or without Infusion of Plasma from Subjects after Convalescence of SARS-CoV-2 Infection in High-Risk Patients with Confirmed Severe SARS-CoV-2 Disease (RECOVER): A structured summary of a study protocol for a randomised controlled trial.

Trials 2020 Oct 6;21(1):828. Epub 2020 Oct 6.

Department of Internal Medicine V, Heidelberg University Hospital, Heidelberg, Germany.

Objectives: Primary objectives • To assess the time from randomisation until an improvement within 84 days defined as two points on a seven point ordinal scale or live discharge from the hospital in high-risk patients (group 1 to group 4) with SARS-CoV-2 infection requiring hospital admission by infusion of plasma from subjects after convalescence of SARS-CoV-2 infection or standard of care. Secondary objectives • To assess overall survival, and the overall survival rate at 28 56 and 84 days. • To assess SARS-CoV-2 viral clearance and load as well as antibody titres. • To assess the percentage of patients that required mechanical ventilation. • To assess time from randomisation until discharge.

Trial Design: Randomised, open-label, multicenter phase II trial, designed to assess the clinical outcome of SARS-CoV-2 disease in high-risk patients (group 1 to group 4) following treatment with anti-SARS-CoV-2 convalescent plasma or standard of care.

Participants: High-risk patients >18 years of age hospitalized with SARS-CoV-2 infection in 10-15 university medical centres will be included. High-risk is defined as SARS-CoV-2 positive infection with Oxygen saturation at ≤ 94% at ambient air with additional risk features as categorised in 4 groups: • Group 1, pre-existing or concurrent hematological malignancy and/or active cancer therapy (incl. chemotherapy, radiotherapy, surgery) within the last 24 months or less. • Group 2, chronic immunosuppression not meeting the criteria of group 1. • Group 3, age ≥ 50 - 75 years meeting neither the criteria of group 1 nor group 2 and at least one of these criteria: Lymphopenia < 0.8 x G/l and/or D-dimer > 1μg/mL. • Group 4, age ≥ 75 years meeting neither the criteria of group 1 nor group 2. Observation time for all patients is expected to be at least 3 months after entry into the study. Patients receive convalescent plasma for two days (day 1 and day 2) or standard of care. For patients in the standard arm, cross over is allowed from day 10 in case of not improving or worsening clinical condition. Nose/throat swabs for determination of viral load are collected at day 0 and day 1 (before first CP administration) and subsequently at day 2, 3, 5, 7, 10, 14, 28 or until discharge. Serum for SARS-Cov-2 diagnostic is collected at baseline and subsequently at day 3, 7, 14 and once during the follow-up period (between day 35 and day 84). There is a regular follow-up of 3 months. All discharged patients are followed by regular phone calls. All visits, time points and study assessments are summarized in the Trial Schedule (see full protocol Table 1). All participating trial sites will be supplied with study specific visit worksheets that list all assessments and procedures to be completed at each visit. All findings including clinical and laboratory data are documented by the investigator or an authorized member of the study team in the patient's medical record and in the electronic case report forms (eCRFs).

Intervention And Comparator: This trial will analyze the effects of convalescent plasma from recovered subjects with SARS-CoV-2 antibodies in high-risk patients with SARS-CoV-2 infection. Patients at high risk for a poor outcome due to underlying disease, age or condition as listed above are eligible for enrollment. In addition, eligible patients have a confirmed SARS-CoV-2 infection and O saturation ≤ 94% while breathing ambient air. Patients are randomised to receive (experimental arm) or not receive (standard arm) convalescent plasma in two bags (238 - 337 ml plasma each) from different donors (day 1, day 2). A cross over from the standard arm into the experimental arm is possible after day 10 in case of not improving or worsening clinical condition.

Main Outcomes: Primary endpoints: The main purpose of the study is to assess the time from randomisation until an improvement within 84 days defined as two points on a seven-point ordinal scale or live discharge from the hospital in high-risk patients (group 1 to group 4) with SARS-CoV-2 infection requiring hospital admission by infusion of plasma from subjects after convalescence of a SARS-CoV-2 infection or standard of care. Secondary endpoints: • Overall survival, defined as the time from randomisation until death from any cause 28-day, 56-day and 84-day overall survival rates. • SARS-CoV-2 viral clearance and load as well as antibody titres. • Requirement mechanical ventilation at any time during hospital stay (yes/no). • Time until discharge from randomisation. • Viral load, changes in antibody titers and cytokine profiles are analysed in an exploratory manner using paired non-parametric tests (before - after treatment).

Randomisation: Upon confirmation of eligibility (patients must meet all inclusion criteria and must not meet exclusion criteria described in section 5.3 and 5.4 of the full protocol), the clinical site must contact a centralized internet randomization system ( https://randomizer.at/ ). Patients are randomized using block randomisation to one of the two arms, experimental arm or standard arm, in a 1:1 ratio considering a stratification according to the 4 risk groups (see Participants).

Blinding (masking): The study is open-label, no blinding will be performed.

Numbers To Be Randomised (sample Size): A total number of 174 patients is required for the entire trial, n=87 per group.

Trial Status: Protocol version 1.2 dated 09/07/2020. A recruitment period of approximately 9 months and an overall study duration of approximately 12 months is anticipated. Recruitment of patients starts in the third quarter of 2020. The study duration of an individual patient is planned to be 3 months. After finishing all study-relevant procedures, therapy, and follow-up period, the patient is followed in terms of routine care and treated if necessary. Total trial duration: 18 months Duration of the clinical phase: 12 months First patient first visit (FPFV): 3 Quarter 2020 Last patient first visit (LPFV): 2 Quarter 2021 Last patient last visit (LPLV): 3 Quarter 2021 Trial Report completed: 4 Quarter 2021 TRIAL REGISTRATION: EudraCT Number: 2020-001632-10, https://www.clinicaltrialsregister.eu/ctr-search/trial/2020-001632-10/DE , registered on 04/04/2020.

Full Protocol: The full protocol is attached as an additional file, accessible from the Trials website (Additional file 1). In the interest in expediting dissemination of this material, the familiar formatting has been eliminated; this Letter serves as a summary of the key elements of the full protocol. The study protocol has been reported in accordance with the Standard Protocol Items: Recommendations for Clinical Interventional Trials (SPIRIT) guidelines (Additional file 2). The eCRF is attached (Additional file 3).
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http://dx.doi.org/10.1186/s13063-020-04735-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7538058PMC
October 2020

Assessment of CAR T Cell Frequencies in Axicabtagene Ciloleucel and Tisagenlecleucel Patients Using Duplex Quantitative PCR.

Cancers (Basel) 2020 Sep 30;12(10). Epub 2020 Sep 30.

Department of Internal Medicine V (Hematology/Oncology/Rheumatology), University Hospital Heidelberg, 69120 Heidelberg, Germany.

Chimeric antigen receptor (CAR) T cell (CART) therapy has been established as a treatment option for patients with CD19-positive lymphoid malignancies in both the refractory and the relapsed setting. Displaying significant responses in clinical trials, two second-generation CART products directed against CD19, axicabtagene ciloleucel (axi-cel) and tisagenlecleucel (tisa-cel), have been approved and integrated into the clinical routine. However, experimental assay for quantitative monitoring of both of these CART products in treated patients in the open domain are lacking. To address this issue, we established and validated a quantitative single copy gene (SCG)-based duplex (DP)-PCR assay (SCG-DP-PCR) to quantify CARTs based on the FMC63 single chain variable fragment (scFv), i.e., axi-cel and tisa-cel. This quantitative PCR (qPCR) approach operates without standard curves or calibrator samples, offers a tool to assess cellular kinetics of FMC63 CARTs and allows direct comparison of CART-copies in axi-cel versus tisa-cel patient samples. For treating physicians, SCG-DP-PCR is an important tool to monitor CARTs and guide clinical decisions regarding CART effects in respective patients.
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http://dx.doi.org/10.3390/cancers12102820DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7601213PMC
September 2020

Biochemical Characterization of Mouse Retina of an Alzheimer's Disease Model by Raman Spectroscopy.

ACS Chem Neurosci 2020 10 8;11(20):3301-3308. Epub 2020 Oct 8.

Leibniz Institute of Photonic Technology (Leibniz-IPHT), a member of the Leibniz Research Alliance Leibniz Health Technology, Albert-Einstein-Straße 9, 07745 Jena, Germany.

The presence of biomarkers characteristic for Alzheimer's disease in the retina is a controversial topic. Raman spectroscopy offers information on the biochemical composition of tissues. Thus, it could give valuable insight into the diagnostic value of retinal analysis. Within the present study, retinas of a double transgenic mouse model, that expresses a chimeric mouse/human amyloid precursor protein and a mutant form of human presenilin 1, and corresponding control group were subjected to Raman imaging. The Raman data recorded on cross sections of whole eyes highlight the layered structure of the retina in a label-free manner. Based on the Raman information obtained from mounted retina samples, a discrimination between healthy and Alzheimer's disease retinal tissue can be done with an accuracy of 85.9%. For this a partial least squares-linear discriminant analysis was applied. Therefore, although no macromolecular changes in form of, , amyloid beta plaques, can be noticed based on Raman spectroscopy, subtle biochemical changes happening in the retina could lead to Alzheimer's disease identification.
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http://dx.doi.org/10.1021/acschemneuro.0c00420DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7581290PMC
October 2020

Comments on "Cost of decentralized CAR T cell production in an academic non-profit setting".

Int J Cancer 2021 01 4;148(2):514-515. Epub 2020 Sep 4.

Department of Internal Medicine V, Hematology, Oncology, Rheumatology, University Hospital Heidelberg, Heidelberg, Germany.

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http://dx.doi.org/10.1002/ijc.33254DOI Listing
January 2021

Cloud removal in Sentinel-2 imagery using a deep residual neural network and SAR-optical data fusion.

ISPRS J Photogramm Remote Sens 2020 Aug;166:333-346

Signal Processing in Earth Observation, Technical University of Munich, Arcisstraße 21, 80333 Munich, Germany.

Optical remote sensing imagery is at the core of many Earth observation activities. The regular, consistent and global-scale nature of the satellite data is exploited in many applications, such as cropland monitoring, climate change assessment, land-cover and land-use classification, and disaster assessment. However, one main problem severely affects the temporal and spatial availability of surface observations, namely cloud cover. The task of removing clouds from optical images has been subject of studies since decades. The advent of the Big Data era in satellite remote sensing opens new possibilities for tackling the problem using powerful data-driven deep learning methods. In this paper, a deep residual neural network architecture is designed to remove clouds from multispectral Sentinel-2 imagery. SAR-optical data fusion is used to exploit the synergistic properties of the two imaging systems to guide the image reconstruction. Additionally, a novel cloud-adaptive loss is proposed to maximize the retainment of original information. The network is trained and tested on a globally sampled dataset comprising real cloudy and cloud-free images. The proposed setup allows to remove even optically thick clouds by reconstructing an optical representation of the underlying land surface structure.
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http://dx.doi.org/10.1016/j.isprsjprs.2020.05.013DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7386944PMC
August 2020