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BackgroundThere is evidence that overexposure to digital media devices (DMD) can not only lead to addictive patterns of internet use, but also cause insomnia symptoms. The aim of this cross-sectional study among adolescents is to provide an estimate of the prevalence of sleep impairments and to explore the mediating role of after-midnight use of DMD between internet-related addictive behavior (IRAB) and insomnia.Methods2,712 school students from Styrian schools participated in a population-representative online survey in a supervised school setting in spring 2022. School students were screened using established and validated scales. Data analysis was carried out using multiple imputation, linear multilevel regression and mediation analysis.ResultsPrevalence estimation indicates high proportions of clinically relevant moderate [12.6% (11.3%; 14.1%)] and severe [3.6% (2.9%; 4.4%)] insomnia, with an additional 30.6% (29.0%; 32.2%) at subthreshold level. DMD are typically used after midnight an average of 1.66 (1.58; 1.75) evenings with subsequent school day per school week. Linear multilevel regression analysis shows significant associations for sleep disparities as outcome variable e.g., with generalized anxiety [b = 0.329 (0.287; 0.371)], after-midnight use of DMD [b = 0.470 (0.369; 0.572)] and IRAB [b = 0.131 (0.097; 0.165)]. Mediation analysis shows a mediated proportion of 18.2% (13.0%; 25.0%) of the association of IRAB and insomnia by after-midnight use of DMD [Indirect effect: b = 0.032 (0.023; 0.040), direct effect: b = 0.127 (0.083; 0.170)].ConclusionsAlthough the cross-sectional nature of this study limits causal inference, the results indicate a need for policies, which are already in preparation in Styria as part of a respective action plan.

MICA and MICB are Class I MHC-related glycoproteins that are upregulated on the surface of cells in response to stress, for instance due to infection or malignant transformation. MICA/B are ligands for NKG2D, an activating receptor on NK cells, CD8+ T cells, and γδ T cells. Upon engagement of MICA/B with NKG2D, these cytotoxic cells eradicate MICA/B-positive targets. MICA is frequently overexpressed on the surface of cancer cells of epithelial and hematopoietic origin. Here, we created nanobodies that recognize MICA. Nanobodies, or VHHs, are the recombinantly expressed variable regions of camelid heavy chain-only immunoglobulins. They retain the capacity of antigen recognition but are characterized by their stability and ease of production. The nanobodies described here detect surface-disposed MICA on cancer cells in vitro by flow cytometry and can be used therapeutically as nanobody-drug conjugates when fused to the Maytansine derivative DM1. The nanobody-DM1 conjugate selectively kills MICA positive tumor cells in vitro.

Summary: Triple-negative breast cancer (TNBC) is a heterogeneous disease with limited treatment options. To characterize TNBC heterogeneity, we defined transcriptional, epigenetic, and metabolic subtypes and subtype-driving super-enhancers and transcription factors by combining functional and molecular profiling with computational analyses. Single-cell RNA sequencing revealed relative homogeneity of the major transcriptional subtypes (luminal, basal, and mesenchymal) within samples. We found that mesenchymal TNBCs share features with mesenchymal neuroblastoma and rhabdoid tumors and that the PRRX1 transcription factor is a key driver of these tumors. PRRX1 is sufficient for inducing mesenchymal features in basal but not in luminal TNBC cells via reprogramming super-enhancer landscapes, but it is not required for mesenchymal state maintenance or for cellular viability. Our comprehensive, large-scale, multiplatform, multiomics study of both experimental and clinical TNBC is an important resource for the scientific and clinical research communities and opens venues for future investigation.

TGFβ signalling is reported to regulate hormone-responsive mammary epithelial progenitors that are associated with breast cancer risk. Here, the authors find that short-term TGFBR1 inhibition prevents tumour formation in rat breast cancer models and identify a TGFBR1 inhibition-responsive sub-population of mammary epithelial cells, which is associated with human breast cancer risk.

Despite the widespread use of online sexual material in the general population, the overlap of deviant use of online sexual material (child sexual abusive material) with other types of online sex and gaming addictions has remained controversial. The purpose of this study is to determine, how users of child sexual abusive material (CSAM) differ from persons consulting for problematic online sex and for gaming (sociodemographic details, Internet use patterns, adverse childhood experiences). We assessed online addiction criteria and adverse childhood experiences in 479 consecutive patients (57 CSAM, 67 users of online sexual material, 355 online gamers) by standardized self-report questionnaires (AICA, CTS). Patients presenting with CSAM consumption were well-educated, socially and professionally integrated middle-aged men. While the quantity of Internet use was less excessive compared to users of online sexual material and online gamers, their average online use was mostly addictive and disruptive regarding partnership or family. They reported significantly higher rates of own experiences of sexual childhood abuse and emotional neglect compared to gamers. Better understanding of CSAM consumption in the context of online addictions is a prerequisite for developing specific therapeutic approaches. These need to consider addictive behavior as well as sustained childhood adversities, which might lead to these maladaptive coping strategies.

The role of MYC in transcriptional reprogramming in prostate cancer remains poorly characterized. Here, MYC overexpression antagonizes the canonical AR transcriptional program leading to prostate tumor initiation and progression by disrupting transcriptional pause release at AR-regulated genes.

The major histocompatibility complex-class I chain related proteins A and B (MICA/B) is upregulated because of cellular stress and MICA/B shedding by cancer cells causes escape from NKG2D recognition favoring the emergence of cancers. Cholangiocarcinoma (CCA) is a relatively rare, though increasingly prevalent, primary liver cancer characterized by a late clinical presentation and a dismal prognosis. We explored the NKG2D-MICA/B axis in NK cells from 41 patients with intrahepatic cholangiocarcinoma (iCCA). The MICA/B-specific 7C6 mAb was used for ex vivo antibody-dependent cytotoxicity (ADCC) experiments using circulating, non tumor liver- and tumor-infiltrating NK cells against the HuCCT-1 cell line and patient-derived primary iCCA cells as targets. MICA/B were more expressed in iCCA than in non-tumoral tissue, MICA transcription being higher in moderately-differentiated compared with poorly-differentiated cancer. Serum MICA was elevated in iCCA patients in line with higher expression of ADAM10 and ADAM17 that are responsible for proteolytic release of MICA/B from tumor. Addition of 7C6 significantly boosted peripheral, liver- and tumor-infiltrating-NK cell degranulation and IFNγ production toward MICA/B-expressing established cell lines and autologous iCCA patient target cells. Our data show that anti-MICA/B drives NK cell anti-tumor activity, and provide preclinical evidence in support of 7C6 as a potential immunotherapeutic tool for iCCA.

Abstract Background Clinical signs and their progression in Beagles with Lafora disease are poorly described. Objectives To describe clinical signs in Beagles with Lafora disease. Animals Twenty‐eight Beagles with Lafora disease confirmed by genetic testing or histopathology. Methods Retrospective multicenter case series. Data regarding signalment, clinical signs, diagnostic tests and treatment were retrieved from hospital data files. A questionnaire was sent to owners asking about neurological deficits, changes in cognitive functions, behavioral changes, response to treatment and survival time. Results Onset of clinical signs was 8.3 years (mean; range, 6.3‐13.3). All dogs had myoclonic episodes as an initial clinical sign with tonic‐clonic seizures in n = 11/28 (39%) and n = 12/28 (43%) later developing tonic‐clonic seizures. Deficits of coordination (n = 21/25; 84%), impaired vision (n = 15/26; 58%), and impaired hearing (n = 13/26; 50%) developed later. Mental decline was observed as loss of house training (urination; n = 8/25; 32%), difficulties performing learned tasks (n = 9/25; 36%), and difficulties learning new tasks (n = 7/23; 30%). Common behavioral changes were: increased photosensitivity (n = 20/26; 77%), staring into space (n = 16/25; 64%), reduced stress resistance (n = 15/26; 58%), increased noise sensitivity (n = 14/26; 54%), and separation anxiety (n = 11/25; 44%). Twenty‐one dogs were alive (median age 11.9 years; range, 9.8‐18.6), and 7 dogs were dead (mean age 12.1 years; SD: 1.3; range, 10.5‐12.6) at time of writing. Conclusions and Clinical Importance Lafora disease in Beagles causes significant behavioral changes, and mental decline as well as neurological deficits in addition to myoclonic episodes and generalized tonic‐clonic seizures. Nevertheless, a relatively normal life span can be expected.

Recent research has shown that risk and reward are positively correlated in many environments, and that people have internalized this association as a “risk-reward heuristic”: when making choices based on incomplete information, people infer probabilities from payoffs and vice-versa, and these inferences shape their decisions. We extend this work by examining people’s expectations about another fundamental trade-off — that between monetary reward and delay. In 2 experiments (total N = 670), we adapted a paradigm previously used to demonstrate the risk-reward heuristic. We presented participants with intertemporal choice tasks in which either the delayed reward or the length of the delay was obscured. Participants inferred larger rewards for longer stated delays, and longer delays for larger stated rewards; these inferences also predicted people’s willingness to take the delayed option. In exploratory analyses, we found that older participants inferred longer delays and smaller rewards than did younger ones. All of these results replicated in 2 large-scale pre-registered studies with participants from a different population (total N = 2138). Our results suggest that people expect intertemporal choice tasks to offer a trade-off between delay and reward, and differ in their expectations about this trade-off. This “delay-reward heuristic” offers a new perspective on existing models of intertemporal choice and provides new insights into unexplained and systematic individual differences in the willingness to delay gratification.

Abstract Background Although gender plays a pivotal role in the psychological adaptation of immigrant youth, its association with acculturation strategy and mental health among 1st generation immigrant adolescents are still scarce and inconsistent. Therefore, the purpose of the current study was to investigate gender-related differences in acculturation patterns and their association with mental health (internalizing and externalizing problems). Methods Self-reported data of immigrant adolescents (N = 440) aged between 12 and 19 years (M = 16.2; SD = 1.6) was collected in a representative German school survey. Fifty-one percent of the sample were female (n = 224). Almost half of the sample was born in the Former Soviet Union, followed by Poland (9.3%). Sociodemographic variables, acculturation strategies, and internalizing as well as externalizing problems were assessed by questionnaires. Results Confirmatory factor analysis supported the four- dimensional model of acculturation styles (assimilation, integration, separation and marginalization). Whereas girls more often showed an integration pattern, boys scored higher on the separation and marginalization scale. After adjusting for age and educational level, regression analyses revealed for both gender that marginalization was associated with more internalizing problems. Separation was related to more externalizing problems. Conclusion 1st generation adolescents experiencing a lack of belongingness to German society, socio-economic and educational disadvantages might be particularly vulnerable to mental distress. Findings are discussed in terms of gender-related differential socialization processes in context of immigration.

Effective combination therapies to improve the efficacy of BET inhibitors are currently under investigation. Here, the authors examine palbociclib and paclitaxel as two promising candidates for combination therapies with BET inhibition in breast cancer and investigate the dynamics of resistance to these combinations through DNA barcoding and mathematical modelling.

We here assess long-term trends in marine primary producers in the southern North Sea (SNS) with respect to ongoing regional Earth system changes. We applied a coupled high-resolution (1.5–4.5 km) 3d-physical-biogeochemical regional Earth System model that includes an advanced phytoplankton growth model and benthic biogeochemistry to hindcast ecosystem dynamics in the period 1961–2012. We analyzed the simulation together with in situ observations. Coinciding with the decreasing nutrient level at the beginning of the 1990s, we find a surprising increase in phytoplankton in the German Bight, but not in the more offshore parts of the SNS. We explain these complex patterns by a series of factors which are lacking in many state-of-the-art coupled ecosystem models such as changed light availability and physiological acclimation in phytoplankton. We also show that many coastal time-series stations in the SNS are located in small patches where our model predicts an opposite trend than found for the surrounding waters. Together, these findings call for a reconsideration of current modeling and monitoring schemes.

The five known New World hemorrhagic fever arenaviruses use the same entry receptor, but their viral glycoproteins are markedly different. Here, the authors isolate monoclonal antibodies from a Junin virus vaccinated person that cross-neutralize Machupo virus and identify a conserved epitope in the receptor-binding domain.

Targeted delivery of immunomodulatory compounds to defined subsets of endogenous immune cells may improve the efficacy of combination immunotherapies. Here, the authors use PD-1-targeting nanoparticles containing a TGFβ inhibitor or a TLR7/8 agonist to deliver these payloads to T cells or via T cells to the tumor microenvironment, respectively, leading to anti-tumor efficacy in vivo.

Abstract Background Published diagnostic questionnaires for gambling disorder in German are either based on DSM-III criteria or focus on aspects other than life time prevalence. This study was designed to assess the usability of the DSM-IV criteria based Berlin Inventory of Gambling Behavior Screening tool in a clinical sample and adapt it to DSM-5 criteria. Methods In a sample of 432 patients presenting for behavioral addiction assessment at the University Medical Center Mainz, we checked the screening tool’s results against clinical diagnosis and compared a subsample of n=300 clinically diagnosed gambling disorder patients with a comparison group of n=132. Results The BIG-S produced a sensitivity of 99.7% and a specificity of 96.2%. The instrument’s unidimensionality and the diagnostic improvements of DSM-5 criteria were verified by exploratory and confirmatory factor analysis as well as receiver operating characteristic analysis. Conclusions The BIG-S is a reliable and valid screening tool for gambling disorder and demonstrated its concise and comprehensible operationalization of current DSM-5 criteria in a clinical setting.

Video gaming is an extremely popular leisure-time activity with more than two billion users worldwide (Newzoo, 2017). However, the media as well as professionals have underscored the potential dangers of excessive video gaming. With the present research, we aimed to shed light on the relation between video gaming and gamers’ psychological functioning. Questionnaires on personality and psychological health as well as video gaming habits were administered to 2,734 individuals (2,377 male, 357 female, Mage = 23.06, SDage = 5.91). Results revealed a medium-sized negative correlation between problematic video gaming and psychological functioning with regard to psychological symptoms, affectivity, coping, and self-esteem. Moreover, gamers’ reasons for playing and their preferred game genres were differentially related to psychological functioning with the most notable findings for distraction-motivated players as well as action game players. Future studies are needed to examine whether these psychological health risks reflect the causes or consequences of video gaming.

Phytoplankton communities are increasingly subject to multiple stressors of natural or anthropogenic origin. The cumulative effect of these stressors, however, may vary considerably from the sum of impacts from individual stressors. Nonlinear effects, such as changes in community traits can either boost up (synergistic) or weaken (antagonistic) single stressors. Despite previous empirical studies and meta analyses on the interaction types of various multiple stressors, a more fundamental understanding of cumulative effects is lacking. To fill this gap, we here propose a new theoretical framework that is centered on the concept of interaction traits and their trade-offs. The framework is applied to a novel size-based plankton model resolving multi-species phytoplankton-nutrients-detritus-zooplankton dynamics within the upper mixed layer. The model is validated using data from a series of outdoor mesocosm experiments. In the direct aftermath of single perturbations that increase net growth rate, here nutrient enrichment and grazer removal, the simulated phytoplankton community undergoes structural changes as visible in altered community traits. These temporal variations explain why the multiple stressor interaction switches from antagonistic to synergistic as compensatory trait variations reduce over time of the experiment. This finding can be generalized within our trait-based explanatory framework to mechanistically assess and predict effects of other stressor combinations and for other organism groups.

One of the most striking patterns at the land-ocean interface is the massive increase of chlorophyll-a (CHL) from continental shelves towards the coast, a phenomenon that is classically linked to physical features. Here I propose that the coastal-offshore CHL gradient in a shallow sea has biological origins related to phytoplankton mortality that are neglected in state-of-the-art biogeochemical models. I integrate a trait-based ecosystem model into a modular coupling framework that is applied to the southern North Sea (SNS). The coupled model very well reproduces daily, seasonal and inter-annual (2000-2014) dynamics and meso-scale patterns in macronutrients, zooplankton biomass, and CHL as observed in situ and by remote sensors. Numerical experiments reveal that coast-offshore CHL gradients may predominantly arise from a trophic effect as resolved by an increase in carnivorous grazing towards shallow waters. This carnivory gradient reflects higher near-coast abundance of juvenile fish and benthic filter feeders. Furthermore, the temporal evolution of CHL can be much affected by viral infection as a fast-responding loss process at intermediate to high phytoplankton concentrations. Viral control in the model also prevents excessive and unrealistic blooms during late spring. Herbivores as often only ecological factor considered for explaining the spatio-temporal phytoplankton distribution are in this study supplemented by pathogens as well as pelagic and benthic carnivores as powerful agents, which are barely represented in current modeling but can mediate physical drivers of coastal ecosystems.

Recent advancement in the immunological understanding of genesis of hepatocellular carcinoma (HCC) has implicated a decline in anti-tumour immunity on the background of chronic inflammatory state of liver parenchyma. The development of HCC involves a network of immunological activity in the tumour microenvironment involving continuous interaction between tumour and stromal cells. The reduction in anti-tumour immunity is secondary to changes in various immune cells and cytokines, and the tumour microenvironment plays a critical role in modulating the process of liver fibrosis, hepatocarcinogenesis, epithelial-mesenchymal transition (EMT), tumor invasion and metastasis. Thus, it is considered as one of primary factor behind the despicable tumour behavior and observed poor survival; along with increased risk of recurrence following treatment in HCC. The primary intent of the present review is to facilitate the understanding of the complex network of immunological interactions of various immune cells, cytokines and tumour cells associated with the development and progression of HCC.

A majority of hepatocellular carcinoma (HCC) develops in the setting of persistent chronic inflammation as immunological mechanisms have been shown to play a vital role in the initiation, growth and progression of tumours. The index review has been intended to highlight ongoing immunological changes in the hepatic parenchyma responsible for the genesis and progression of HCC. The in-situ vaccine effect of radiofrequency (RF) is through generation tumour-associated antigens (TAAs), following necrosis and apoptosis of tumour cells, which not only re-activates the antitumour immune response but can also act in synergism with checkpoint inhibitors to generate a superlative effect with intent to treat primary cancer and distant metastasis. An improved understanding of oncogenic responses of immune cells and their integration into signaling pathways of the tumour microenvironment will help in modulating the antitumour immune response. Finally, we analyzed contemporary literature and summarised the recent advances made in the field of targeted immunotherapy involving checkpoint inhibitors along with RF application with the intent to reinstate antitumour immunity and outline future directives in very early and early stages of HCC.

We isolated 65 rhizobial strains from faba bean (Vicia faba L.) from Panxi, China, studied their plant growth promoting ability with nitrogen free hydroponics, genetic diversity with clustered analysis of combined ARDRA and IGS-RFLP, and phylogeny by sequence analyses of 16S rRNA gene, three housekeeping genes and symbiosis related genes. Eleven strains improved the plant shoot dry mass significantly comparing to that of not inoculated plants. According to the clustered analysis of combined ARDRA and IGS-RFLP the isolates were genetically diverse. Forty-one of 65 isolates represented Rhizobium anhuiense, and the others belonged to R. fabae, Rhizobium vallis, Rhizobium sophorae, Agrobacterium radiobacter, and four species related to Rhizobium and Agrobacterium. The isolates carried four and five genotypes of nifH and nodC, respectively, in six different nifH-nodC combinations. When looking at the species-nifH-nodC combinations it is noteworthy that all but two of the six R. anhuiense isolates were different. Our results suggested that faba bean rhizobia in Panxi are diverse at species, plant growth promoting ability and symbiosis related gene levels.

Adolescents with a strong tendency for irrational task delay (i.e., high trait procrastination) may be particularly prone to use Internet applications simultaneously to other tasks (e.g., during homework) and in an insufficiently controlled fashion. Both Internet multitasking and insufficiently controlled Internet usage may thus amplify the negative mental health implications that have frequently been associated with trait procrastination. The present study explored this role of Internet multitasking and insufficiently controlled Internet use for the relationship between trait procrastination and impaired psychological functioning in a community sample of N = 818 early and middle adolescents. Results from multiple regression analyses indicate that trait procrastination was positively related to Internet multitasking and insufficiently controlled Internet use. Insufficiently controlled Internet use, but not Internet multitasking, was found to partially statistically mediate the association between trait procrastination and adolescents’ psychological functioning (i.e., stress, sleep quality, and relationship satisfaction with parents). The study underlines that adolescents with high levels of trait procrastination may have an increased risk for negative outcomes of insufficiently controlled Internet use.

ABSTRACT This study demonstrates that the deltaproteobacterium Desulfurivibrio alkaliphilus can grow chemolithotrophically by coupling sulfide oxidation to the dissimilatory reduction of nitrate and nitrite to ammonium. Key genes of known sulfide oxidation pathways are absent from the genome of D. alkaliphilus. Instead, the genome contains all of the genes necessary for sulfate reduction, including a gene for a reductive-type dissimilatory bisulfite reductase (DSR). Despite this, growth by sulfate reduction was not observed. Transcriptomic analysis revealed a very high expression level of sulfate-reduction genes during growth by sulfide oxidation, while inhibition experiments with molybdate pointed to elemental sulfur/polysulfides as intermediates. Consequently, we propose that D. alkaliphilus initially oxidizes sulfide to elemental sulfur, which is then either disproportionated, or oxidized by a reversal of the sulfate reduction pathway. This is the first study providing evidence that a reductive-type DSR is involved in a sulfide oxidation pathway. Transcriptome sequencing further suggests that nitrate reduction to ammonium is performed by a novel type of periplasmic nitrate reductase and an unusual membrane-anchored nitrite reductase. IMPORTANCE Sulfide oxidation and sulfate reduction, the two major branches of the sulfur cycle, are usually ascribed to distinct sets of microbes with distinct diagnostic genes. Here we show a more complex picture, as D. alkaliphilus, with the genomic setup of a sulfate reducer, grows by sulfide oxidation. The high expression of genes typically involved in the sulfate reduction pathway suggests that these genes, including the reductive-type dissimilatory bisulfite reductases, are also involved in as-yet-unresolved sulfide oxidation pathways. Finally, D. alkaliphilus is closely related to cable bacteria, which grow by electrogenic sulfide oxidation. Since there are no pure cultures of cable bacteria, D. alkaliphilus may represent an exciting model organism in which to study the physiology of this process.

Autotrophic organisms reveal an astounding flexibility in their elemental stoichiometry, with potentially major implications on biogeochemical cycles and ecological functioning. Notwithstanding, stoichiometric regulation and co-limitation by multiple resources in autotrophs revt were in the past often described by heuristic formulations.In this study, we present a mechanistic model of autotroph growth, which features two major improvements over the existing schemes. First, we introduce the concept of metabolic network independence that defines the degree of phase-locking between accessory machines. Network independence is in particular suggested to be proportional to protein synthesis capability as quantified by variable intracellular N:C. Consequently, the degree of co-limitation becomes variable, contrasting with the dichotomous debate on the use of Liebig's law or the product rule, standing for constantly low and high co-limitation, respectively. Second, we resolve dynamic protein partitioning to light harvesting, carboxylation processes, and to an arbitrary number of nutrient acquisition machineries, as well as instantaneous activity regulation of nutrient uptake. For all regulatory processes we assume growth rate optimality, here extended by an explicit consideration of indirect feed-back effects.The combination of network independence and optimal regulation displays unprecedented skill in reproducing rich stoichiometric patterns collected from a large number of published chemostat experiments. This high skill indicates (1) that the current paradigm of fixed co-limitation is a critical short-coming of conventional models, and (2) that stoichiometric flexibility in autotrophs possibly reflects an optimality strategy. Numerical experiments furthermore show that regulatory mechanisms homogenize the effect of multiple stressors. Extended optimality alleviates the effect of the most limiting resource(s) while down-regulating machineries for the less limiting ones, which induces an ubiquitous response surface of growth rate over ambient resource levels.

Addressing the lack of population-based data the purpose of this representative study was to assess procrastination and its associations with distress and life satisfaction across the life span. A representative German community sample (1,350 women; 1,177 men) between the ages of 14 and 95 years was examined by the short form of the General Procrastination Scale (GPS-K; 1) and standardized scales of perceived stress, depression, anxiety, fatigue and life satisfaction. As hypothesized, procrastination was highest in the youngest cohort (14-29 years). Only in the youngest and most procrastinating cohort (aged 14 to 29 years), men procrastinated more than women. As we had further hypothesized, procrastination was consistently associated with higher stress, more depression, anxiety, fatigue and reduced satisfaction across life domains, especially regarding work and income. Associations were also found with lack of a partnership and unemployment. Findings are discussed with regard to potential developmental and cohort effects. While procrastination appears to be a pervasive indicator for maladjustment, longitudinal analyses in high-risk samples (e.g. late adolescence, unemployment) are needed to identify means and mechanisms of procrastinating.

ObjectiveWe aimed to develop a distributed, immutable, and highly available cross-cloud blockchain system to facilitate federated data analysis activities among multiple institutions.Materials and methodsWe preprocessed 9166 COVID-19 Structured Query Language (SQL) code, summary statistics, and user activity logs, from the GitHub repository of the Reliable Response Data Discovery for COVID-19 (R2D2) Consortium. The repository collected local summary statistics from participating institutions and aggregated the global result to a COVID-19-related clinical query, previously posted by clinicians on a website. We developed both on-chain and off-chain components to store/query these activity logs and their associated queries/results on a blockchain for immutability, transparency, and high availability of research communication. We measured run-time efficiency of contract deployment, network transactions, and confirmed the accuracy of recorded logs compared to a centralized baseline solution.ResultsThe smart contract deployment took 4.5 s on an average. The time to record an activity log on blockchain was slightly over 2 s, versus 5-9 s for baseline. For querying, each query took on an average less than 0.4 s on blockchain, versus around 2.1 s for baseline.DiscussionThe low deployment, recording, and querying times confirm the feasibility of our cross-cloud, blockchain-based federated data analysis system. We have yet to evaluate the system on a larger network with multiple nodes per cloud, to consider how to accommodate a surge in activities, and to investigate methods to lower querying time as the blockchain grows.ConclusionBlockchain technology can be used to support federated data analysis among multiple institutions.

Chronic lymphocytic leukemia (CLL) is thought to be a disease of resting lymphocytes. However, recent data suggest that CLL cells may more closely resemble activated B cells. Using microRNA (miRNA) expression profiling of highly-enriched CLL cells from 38 patients and 9 untransformed B cells from normal donors before acute CpG activation and 5 matched B cells after acute CpG activation, we demonstrate an activated B cell status for CLL. Gene set enrichment analysis (GSEA) identified statistically-significant similarities in miRNA expression between activated B cells and CLL cells including upregulation of miR-34a, miR-155, and miR-342-3p and downregulation of miR-103, miR-181a and miR-181b. Additionally, decreased levels of two CLL signature miRNAs miR-29c and miR-223 are associated with ZAP70(+) and IgV(H) unmutated status and with shorter time to first therapy. These data indicate an activated B cell status for CLL cells and suggest that the direction of change of individual miRNAs may predict clinical course in CLL.

The majority of receptors responsible for activation of distinct cell types within the immune system assemble with dimeric signaling modules through interaction of a basic transmembrane residue with a pair of acidic residues of the signaling dimer. Because assembly of other membrane proteins requires specific interactions along extended stretches of transmembrane helices, we examined how transmembrane sequences flanking the polar interaction site contribute to assembly for three receptors that associate with different signaling modules-the natural killer cell receptors KIR and NKG2D and the Fc receptor for IgA, FcalphaRI. The KIR and NKG2D receptors assembled with the DAP12 and DAP10 dimers, respectively, even when the entire KIR or NKG2D transmembrane domains were replaced by polyleucine sequences with a properly positioned basic residue. In contrast, a high degree of specificity for the basic side chain could be observed because the KIR-DAP12 and FcalphaRI-Fcgamma interactions favored lysine or arginine, respectively. Steric hindrance among incompatible extra-membranous domains and competition for signaling modules also contributed to specificity of assembly. These results demonstrate that these interactions are focused on the polar site created by three ionizable transmembrane residues, and explain how the DAP12 and Fcgamma signaling modules can assemble with large, non-overlapping sets of receptors that have highly divergent transmembrane sequences.

CDK4/6 inhibitors are approved to treat breast cancer and are in trials for other malignancies. We examined CDK4/6 inhibition in mouse and human CD8+ T cells during early stages of activation. Mice receiving tumor-specific CD8+ T cells treated with CDK4/6 inhibitors displayed increased T-cell persistence and immunologic memory. CDK4/6 inhibition upregulated MXD4, a negative regulator of MYC, in both mouse and human CD8+ T cells. Silencing of Mxd4 or Myc in mouse CD8+ T cells demonstrated the importance of this axis for memory formation. We used single-cell transcriptional profiling and T-cell receptor clonotype tracking to evaluate recently activated human CD8+ T cells in patients with breast cancer before and during treatment with either palbociclib or abemaciclib. CDK4/6 inhibitor therapy in humans increases the frequency of CD8+ memory precursors and downregulates their expression of MYC target genes, suggesting that CDK4/6 inhibitors in patients with cancer may augment long-term protective immunity. SIGNIFICANCE: CDK4/6 inhibition skews newly activated CD8+ T cells toward a memory phenotype in mice and humans with breast cancer. CDK4/6 inhibitors may have broad utility outside breast cancer, particularly in the neoadjuvant setting to augment CD8+ T-cell priming to tumor antigens prior to dosing with checkpoint blockade.This article is highlighted in the In This Issue feature, p. 2355.

ObjectiveTo utilize, in an individual and institutional privacy-preserving manner, electronic health record (EHR) data from 202 hospitals by analyzing answers to COVID-19-related questions and posting these answers online.Materials and methodsWe developed a distributed, federated network of 12 health systems that harmonized their EHRs and submitted aggregate answers to consortia questions posted at https://www.covid19questions.org. Our consortium developed processes and implemented distributed algorithms to produce answers to a variety of questions. We were able to generate counts, descriptive statistics, and build a multivariate, iterative regression model without centralizing individual-level data.ResultsOur public website contains answers to various clinical questions, a web form for users to ask questions in natural language, and a list of items that are currently pending responses. The results show, for example, that patients who were taking angiotensin-converting enzyme inhibitors and angiotensin II receptor blockers, within the year before admission, had lower unadjusted in-hospital mortality rates. We also showed that, when adjusted for, age, sex, and ethnicity were not significantly associated with mortality. We demonstrated that it is possible to answer questions about COVID-19 using EHR data from systems that have different policies and must follow various regulations, without moving data out of their health systems.Discussion and conclusionsWe present an alternative or a complement to centralized COVID-19 registries of EHR data. We can use multivariate distributed logistic regression on observations recorded in the process of care to generate results without transferring individual-level data outside the health systems.

T cells are critical effectors of cancer immunotherapies, but little is known about their gene expression programs in diffuse gliomas. Here, we leverage single-cell RNA sequencing (RNA-seq) to chart the gene expression and clonal landscape of tumor-infiltrating T cells across 31 patients with isocitrate dehydrogenase (IDH) wild-type glioblastoma and IDH mutant glioma. We identify potential effectors of anti-tumor immunity in subsets of T cells that co-express cytotoxic programs and several natural killer (NK) cell genes. Analysis of clonally expanded tumor-infiltrating T cells further identifies the NK gene KLRB1 (encoding CD161) as a candidate inhibitory receptor. Accordingly, genetic inactivation of KLRB1 or antibody-mediated CD161 blockade enhances T cell-mediated killing of glioma cells in vitro and their anti-tumor function in vivo. KLRB1 and its associated transcriptional program are also expressed by substantial T cell populations in other human cancers. Our work provides an atlas of T cells in gliomas and highlights CD161 and other NK cell receptors as immunotherapy targets.

Our knowledge of cancer genomics exploded in last several years, providing us with detailed knowledge of genetic alterations in almost all cancer types. Analysis of this data gave us new insights into molecular aspects of cancer, most important being the amazing diversity of molecular abnormalities in individual cancers. The most important question in cancer research today is how to classify this diversity to identify subtypes that are most relevant for treatment and outcome prediction for individual patients. The Cancer3D database at http://www.cancer3d.org gives an open and user-friendly way to analyze cancer missense mutations in the context of structures of proteins they are found in and in relation to patients' clinical data. This approach allows users to find novel candidate driver regions for specific subgroups, that often cannot be found when similar analyses are done on the whole gene level and for large, diverse cohorts. Interactive interface allows user to visualize the distribution of mutations in subgroups defined by cancer type and stage, gender and age brackets, patient's ethnicity or vice versa find dominant cancer type, gender or age groups for specific three-dimensional mutation patterns.