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BACKGROUND: Trigeminal neuralgia (TN) is a severe facial pain condition often associated with a neurovascular conflict. However, neuroinflammation has also been implicated in TN, as it frequently co-occurs with multiple sclerosis (MS).

METHODS: We analysed protein expression levels of TN patients compared to MS patients and controls. Proximity Extension Assay technology was used to analyse the levels of 92 proteins with the Multiplex Neuro-Exploratory panel provided by SciLifeLab, Uppsala, Sweden. Serum and CSF samples were collected from TN patients before (n = 33 and n = 27, respectively) and after (n = 28 and n = 8, respectively) microvascular decompression surgery. Additionally, we included samples from MS patients (n = 20) and controls (n = 20) for comparison.

RESULTS: In both serum and CSF, several proteins were found increased in TN patients compared to either MS patients, controls, or both, including EIF4B, PTPN1, EREG, TBCB, PMVK, FKBP5, CD63, CRADD, BST2, CD302, CRIP2, CCL27, PPP3R1, WWP2, KLB, PLA2G10, TDGF1, SMOC1, RBKS, LTBP3, CLSTN1, NXPH1, SFRP1, HMOX2, and GGT5. The overall expression of the 92 proteins in postoperative TN samples seems to shift towards the levels of MS patients and controls in both serum and CSF, as compared to preoperative samples. Interestingly, there was no difference in protein levels between MS patients and controls.

CONCLUSIONS: We conclude that TN patients showed increased serum and CSF levels of specific proteins and that successful surgery normalizes these protein levels, highlighting its potential as an effective treatment. However, the similarity between MS and controls challenges the idea of shared pathophysiology with TN, suggesting distinct underlying mechanisms in these conditions.

SIGNIFICANCE: This study advances our understanding of trigeminal neuralgia (TN) and its association with multiple sclerosis (MS). By analysing 92 protein biomarkers, we identified distinctive molecular profiles in TN patients, shedding light on potential pathophysiological mechanisms. The observation that successful surgery normalizes many protein levels suggests a promising avenue for TN treatment. Furthermore, the contrasting protein patterns between TN and MS challenge prevailing assumptions of similarity between the two conditions and point to distinct pathophysiological mechanisms.

Introduction Depression is a major global burden with unclear pathophysiology and poor treatment outcomes. Diagnosis of depression continues to rely primarily on behavioral rather than biological methods. Investigating tools that might aid in diagnosing and treating early-onset depression is essential for improving the prognosis of the disease course. While there is increasing evidence of possible biomarkers in adult depression, studies investigating this subject in adolescents are lacking.Methods In the current study, we analyzed protein levels in 461 adolescents assessed for depression using the Development and Well-Being Assessment (DAWBA) questionnaire as part of the domestic Psychiatric Health in Adolescent Study conducted in Uppsala, Sweden. We used the Proseek Multiplex Neuro Exploratory panel with Proximity Extension Assay technology provided by Olink Bioscience, followed by transcriptome analyses for the genes corresponding to the significant proteins, using four publicly available cohorts.Results We identified a total of seven proteins showing different levels between DAWBA risk groups at nominal significance, including RBKS, CRADD, ASGR1, HMOX2, PPP3R1, CD63, and PMVK. Transcriptomic analyses for these genes showed nominally significant replication of PPP3R1 in two of four cohorts including whole blood and prefrontal cortex, while ASGR1 and CD63 were replicated in only one cohort.Discussion Our study on adolescent depression revealed protein-level and transcriptomic differences, particularly in PPP3R1, pointing to the involvement of the calcineurin pathway in depression. Our findings regarding PPP3R1 also support the role of the prefrontal cortex in depression and reinforce the significance of investigating prefrontal cortex-related mechanisms in depression.

Trigeminal neuralgia (TN) is a severe facial pain disease of uncertain pathophysiology and unclear genetic background. Although recent research has reported a more important role of genetic factors in TN pathogenesis, few candidate genes have been proposed to date. The present study aimed to identify independent genetic variants in the protein-coding genes associated with TN. We focused on genes previously linked to TN based on the results of four proteomic studies conducted by our research team. The goal was to validate these findings on the genetic level to enhance our understanding of the role of genetics in TN. The study is based on the participants from UK Biobank cohort. Following quality control, 175 independent single nucleotide polymorphisms (SNPs) in 17 genes were selected. The study sample comprised of diagnosed TN cases (N = 555) and randomly matched controls (N = 6245) based on specific criteria. Two SNPs corresponding to C8B rs706484 [odds ratio (OR) (95% confidence interval (CI)): 1.357 (1.158–1.590); p: 0.00016] and MFG-E8 rs2015495 [OR (95% CI): 1.313 (1.134–1.521); p: 0.00028] showed significant positive association with TN, indicating a positive effect of the SNP alleles on gene expression and disease risk. Interestingly, both SNPs are Expression Quantitative Trait Loci (eQTLs), and are associated with changes in the expression activity of their corresponding gene. Our findings suggest novel genetic associations between C8B, a key component of the complement system, and MFG-E8, which plays a role in regulating neuroinflammation, in relation to TN. The identified genetic variations may help explain why some individuals develop TN while others do not, indicating a potential genetic predisposition to the condition.

Fetal growth restriction is a strong risk factor for perinatal morbidity and mortality. Reliable standards are indispensable, both to assess fetal growth and to evaluate birthweight and early postnatal growth in infants born preterm. The aim of this study was to create updated Swedish reference ranges for estimated fetal weight (EFW) from gestational week 12-42. This prospective longitudinal multicentre study included 583 women without known conditions causing aberrant fetal growth. Each woman was assigned a randomly selected protocol of five ultrasound scans from gestational week 12 + 3 to 41 + 6. Hadlock's 3rd formula was used to estimate fetal weight. A two-level hierarchical regression model was employed to calculate the expected median and variance, expressed in standard deviations and percentiles, for EFW. EFW was higher for males than females. The reference ranges were compared with the presently used Swedish, and international reference ranges. Our reference ranges had higher EFW than the presently used Swedish reference ranges from gestational week 33, and higher median, 2.5th and 97.5th percentiles from gestational week 24 compared with INTERGROWTH-21st. The new reference ranges can be used both for assessment of intrauterine fetal weight and growth, and early postnatal growth in children born preterm.

Objective: To investigate fetal growth trajectories and risks of small and large for gestational age (SGA and LGA), and macrosomia in pregnancies after fresh and frozen embryo transfer (ET), and natural conception (NC).

Design: Longitudinal population-based cohort study.

Setting: Swedish national registers.

Population: A total of 196 008 singleton pregnancies between 2013 and 2017.

Methods: Of all singleton pregnancies resulting in live births in the Swedish Pregnancy Register, 10 970 fresh ET, 6520 frozen ET, and 178 518 NC pregnancies with ultrasound data were included. A general least squares model was used to examine the effect of fresh or frozen ET on fetal growth while adjusting for confounders.

Main Outcome: MeasuresFetal growth velocity. SGA, LGA and macrosomia.

Results: At 120 days, fetal weights were lower in fresh ET pregnancies compared with NC pregnancies. Thereafter fresh ET as well as FET fetuses had higher fetal weights than NC fetuses, with no differences between themselves until the second trimester. From 210 days, FET fetuses were heavier than fresh ET fetuses, whereas fresh ET fetuses had lower fetal weights than NC fetuses from 245 days. After fresh ET, SGA was more frequent, whereas LGA and macrosomia were less frequent, than after FET.

Conclusions: This study gives new insights into the differences in fetal growth dynamics between fresh and frozen ET and NC pregnancies. Clinically relevant differences in proportions of SGA, LGA and macrosomia were observed.

This article presents a new, monthly updated dataset on organized violence—the Uppsala Conflict Data Program Candidate Events Dataset. It contains recent observations of candidate events, a majority of which are eventually included in the Uppsala Conflict Data Program Georeferenced Event Dataset as part of its annual update after a careful vetting process. We describe the definitions, sources and procedures employed to code the candidate events, and a set of issues that emerge when coding data on organized violence in near-real time. Together, the Uppsala Conflict Data Program Candidate and Georeferenced Event Datasets minimize an inherent trade-off between update speed and quality control. Having monthly updated conflict data is advantageous for users needing near-real time monitoring of violent situations and aiming to anticipate future developments. To demonstrate this, we show that including them in a conflict forecasting system yields distinct improvements in terms of predictive performance: Average precision increases by 20–40% relative to using the Uppsala Conflict Data Program Georeferenced Event Dataset only. We also show that to ensure quality and consistency, revisiting the initial coding making use of sources that become available later is absolutely necessary.

The proliferation of large-scale, geographically disaggregated data on armed conflicts, protests, and similar events has opened new avenues of research, but has also introduced significant data quality challenges. A notable yet often overlooked issue involves observations with “known geographic imprecision” (KGI), where event locations are unknown and instead arbitrarily assigned by dataset authors. Although this issue is widely recognized and accounts for up to a quarter of observations in datasets like UCDP GED, it is rarely addressed by users. This paper presents a stochastic method derived from the multiple-imputation literature, employing spatio-temporal Gaussian processes and leveraging latent actor-conflict features in the data to enhance location accuracy. Extensive Monte-Carlo simulations demonstrate that this approach substantially enhances the accuracy of these observations and improves predictive performance beyond the state-of-the-art when applied out-of-sample. Additionally, an adapted version of the UCDP GED dataset that employs this new procedure is provided, showcasing the practical application and benefits of the methodology.

High-resolution event data on armed conflict and related processes have revolutionized the study of political contention. However, most datasets of this type only collect spatio-temporal and conflict intensity data at that level of detail. Information on dynamics, such as targets, tactics, and purposes, is rarely collected due to the substantial effort of collecting data. This study proposes an inexpensive, high-performance approach to increase the feature richness of such datasets by leveraging active learning -- an iterative process of improving a machine learning model based on guided human input at each step of the learning process. Active learning is employed to then fine-tune (train in steps) a large, encoder-only language model fitted to the rich corpus of textual data underlying such datasets. This allows for the extraction of features related to conflict dynamics, such as electoral violence and attacks on religious targets. The approach achieves a performance comparable to the human (gold-standard) coding, while reducing the necessary human annotation by as much as 99 percent.

What explains conflict escalation during civil war? This article explores whether provocative attacks on religious sites and public transport constitute a precursor to a surge of violence. One argument pertains that the symbolic and doctrinal importance of places of worship means that attacks on these will affect individuals and the community emotionally and thereby increase the risk of escalation. However, it can also be suggested that the everyday societal importance of a public space is similar for religious sites and public transport hubs. We test these arguments using novel new global event data on these forms of selective targeting for 1989-2015, and find that the risk of conflict escalation increase in the aftermath of either attacks on places of worship or public transport, suggesting that community behavior is more affected to disruptions of societal everyday life than to the importance of symbols.

This study advances the field of conflict forecasting by using text-based actor embeddings with transformer models to predict dynamic changes in violent conflict patterns at the actor level. More specifically, we combine newswire texts with structured conflict event data and leverage recent advances in Natural Language Processing (NLP) techniques to forecast escalations and de-escalations among conflicting actors, such as governments, militias, separatist movements, and terrorists. This new approach accurately and promptly captures the inherently volatile patterns of violent conflicts, which existing methods have not been able to achieve. To create this framework, we began by curating and annotating a vast international newswire corpus, leveraging hand-labeled event data from the Uppsala Conflict Data Program. By using this hybrid dataset,  our models can incorporate the textual context of news sources along with the precision and detail of structured event data. This combination enables us to make both dynamic and granular predictions about conflict developments. We validate our approach through rigorous back-testing against historical events, demonstrating superior out-of-sample predictive power. We find that our approach is quite effective in identifying and predicting phases of conflict escalation and de-escalation, surpassing the capabilities of traditional models. By focusing on actor interactions, our explicit goal is to provide actionable insights to policymakers, humanitarian organizations, and peacekeeping operations in order to enable targeted and effective intervention strategies.

Temporally and spatial disaggregated datasets are commonly used to study political violence. Researchers are increasingly studying the data generation process itself to understand the selection processes by which conflict events are included in conflict datasets. This work has focused on conflict fatalities. In this research note, we explore how non-fatal conflict events are reported upon and enter into datasets of armed conflict. To do so, we compare reported non-fatal conflict events with the population of events in two direct observation datasets, collected using a boots-on-the-ground strategy: mass abductions in Nepal (1996-2006) and troop movements in Darfur. We show that at the appropriate level of aggregation media reporting on abductions in Nepal largely mirrors the "true" population of abductions, but at more disaggregated levels of temporal or spatial analysis, the match is poor. We also show that there is no overlap between a media-driven conflict dataset and directly-observed data on troop movements in Sudan. These empirics indicate that non-fatal data can suffer from serious underreporting and that this is particularly the case for events lacking elements of coercion. These findings are indicative of selection problems in regards to the reporting on non-fatal conflict events.

Modern glucose sensors deployed in closed -loop insulin delivery systems, so-called artificial pancreas use wireless communication channels. While this allows a flexible system design, it also introduces vulnerability to cyberattacks. Timely detection and mitigation of attacks are imperative for device safety. However, large unknown meal disturbances are a crucial challenge in determining whether the sensor has been compromised or the sensor glucose trajectories are normal. We address this issue from a control -theoretic security perspective. In particular, a time -varying Kalman filter is employed to handle the sporadic meal intakes. The filter prediction error is then statistically evaluated to detect anomalies if present. We compare two state-of-the-art online anomaly detection algorithms, namely the ￜￜￜￜￜￜ2 and CUSUM tests. We establish a robust optimal detection rule for unknown bias injections. Even if the optimality holds only for the restrictive case of constant bias injections, we show that the proposed model -based anomaly detection scheme is also effective for generic non -stealthy sensor deception attacks through numerical simulations.

This paper considers deterministic sensor deception attacks in closed-loop insulin delivery. Since the quality of decision-making in control systems heavily relies on accurate sensor measurements, timely detection of attacks is imperative. To this end, we consider a model-based anomaly detection scheme based on Kalman filtering and sequential change detection. In particular, we derive the minimax robust CUSUM and Shewhart tests that minimizes the worst-case mean detection delay and maximizes the instant detection rate, respectively. As a byproduct of our analysis, we show that the notorious.2 test shares an interesting optimality property with the twosided Shewhart test. Finally, we show that one-sided sequential detectors can significantly improve sensor anomaly detection for preventing overnight hypoglycemia which can be fatal.

This paper considers observer-based detection of sensor bias injection attacks (BIAs) on linear cyber-physical systems with single output driven by Gaussian noise. Despite their simplicity, BIAs pose a severe risk to systems with integrators, which we refer to as integrator vulnerability. Specifically, the residual generated by any linear observer is indistinguishable under attack and normal operation at steady-state, making BIAs detectable only during transients. To address this, we propose a principled method based on Kullback-Liebler divergence to design a residual generator that significantly increases the signal-to-noise ratio against BIAs. For systems without integrator vulnerability, our method also enables a trade-off between transient and steady-state detectability. The effectiveness of the proposed method is demonstrated through numerical comparisons with three state-of-the-art residual generators.

Introduction: Homelessness among women of reproductive age is a globalproblem. Several unique gender-based issues affect homeless women’s wellbeing, including reproductive health issues, their homelessness experiences,and a high rate of sexual violence. In this study, we aimed to describe women’sexperiences of street homelessness in their own terms and their suggestions toaddress their unmet needs.Methods: This photovoice study draws on photos, focus group discussions, andsemi-structured interviews. We conducted the study in collaboration with womenin their reproductive years experiencing homelessness (n = 9). A total of 80 photoswere taken, and 40 were chosen to be discussed in interviews and further focusgroup discussions. The participating women selected photographs, explained theirsignificance, and codified them based on how they related to their lives. Data fromthese discussions were then analysed using a reflexive thematic approach.Results: Four themes were developed from the data: (a) deprivation of basicneeds; (b) experiencing dependency, shame, and seclusion while dealingwith the burden of street life; (c) the vulnerability and neglect of children; and(d) being resilient to harsh conditions. In this study, women’s street life wascharacterised by numerous unfavourable aspects, including unmet needs,human rights violations, social exclusions, substance use, and child protectionissues. Participants provided suggestions for change and confirmed their beliefthat adequate housing represents one of the most urgent unmet basic needsof people experiencing homelessness. They also emphasised the critical needfor employment opportunities, non-discriminatory provision of social support,treatment programs for substance misuse, and legal and social protection.Conclusion: This study contributes to understanding how women experiencinghomelessness describe and articulate their living circumstances and whatthey perceive needs to be addressed. Based on participants’ proposal forchange, comprehensive services are needed to address women’s multifacetedissues. However, the mitigation strategies and long-term effects of women’shomelessness require further research.

Background: Studies show that women’s trajectories through homelessness are shaped by gendered individual susceptibility, dysfunctional family dynamics, and social norms, as well as high rates of violence. Although women’s homelessness, specifically that of single mothers, often has a significant impact on the lives of women and their children, there is a dearth of research on this area in Ethiopia. Therefore, the purpose of this study was to explore homeless women’s early life experiences, their trajectories through homelessness, and the challenges inherent in community reintegration, as they perceive them. 

Method: We conducted a qualitative study in December 2023 using semi-structured interviews. We purposively selected homeless women of reproductive age from the beneficiaries of a local civil society in Addis Ababa, Ethiopia (n = 19). 

Results: ‘Childhood trauma from abuse’, ‘Sexual violence’, ‘Barriers to leaving street living, and ‘Sources of hope’ were four themes identified from the data. Despite differences in the reasons for homelessness and the routes that led the women to the streets, most of them experienced abuse, neglect, and exploitation in their early years of life. Rather than giving them a window into healing from their childhood trauma, the street further exposed women to sexual abuse, addiction, and underserved justice. Although there were personal, economic, and normative impediments, some women highlighted how they accepted the situation, looked for support, and relied on their strength and faith. 

Conclusions: The findings highlight how the trajectories through homelessness were shaped by abuse as the common thread in the lived experiences of homeless women in Addis Ababa, Ethiopia. Given the highly gendered and traumatic nature of their situation, our results call for action to address the need for contextualizing tailored and gender-responsive, multi-level trauma-focused domestic violence and sexual violence interventions and rehabilitation services, as well as reintegration mechanism for homeless women of reproductive age.

Background: Mental health conditions are among the health issues associated with homelessness, and providing mental healthcare to people experiencing homelessness is challenging. Despite the pressing issue of homelessness in Addis Ababa, Ethiopia, there is scant research on how service providers address women's mental health and psychosocial needs. Therefore, we explored service providers' and programme coordinators' perceptions and experiences regarding mental healthcare and psychosocial services delivery to women experiencing street homelessness in the city.

Methods: We conducted a descriptive qualitative study with selected healthcare and social support providers and programme coordinators. The study involved 34 participants from governmental and non-governmental organisations in Addis Ababa, Ethiopia. Data were analysed using an inductive thematic approach.

Results: Four themes were derived from the analysis. The first of these was "divergent intentions and actions". While service providers and programme coordinators showed empathy and compassion, they also objectified and blamed people for their own homelessness. They also expressed opposing views on mental health stigma and compassion for these people. The second theme addressed "problem-solution incompatibility", which focused on the daily challenges of women experiencing homelessness and the types of services participants prioritised. Service providers and programme coordinators proposed non-comprehensive support despite the situation's complexity. The participants did not emphasise the significance of gender-sensitive and trauma-informed care for women experiencing street homelessness in the third theme, "the lack of gendered and trauma-informed care despite an acknowledgement that women face unique challenges". The fourth theme, "mismatched resources," indicated structural and systemic barriers to providing services to homeless women.

Conclusions:Conflicting attitudes and practices exist at the individual, organisational, and systemic levels, making it challenging to provide mental healthcare and psychosocial services to women experiencing homelessness. An integrated, gender-sensitive, and trauma-informed approach is necessary to assist women experiencing homelessness.

Introduction: Globally, homelessness is a growing concern, and homeless women of reproductive age are particularly vulnerable to adverse physical, mental, and reproductive health conditions, including violence. Although Ethiopia has many homeless individuals, the topic has received little attention in the policy arena. Therefore, we aimed to understand the reason for the lack of attention, with particular emphasis on women of reproductive age.

Methods: This is a qualitative study; 34 participants from governmental and non-governmental organisations responsible for addressing homeless individuals' needs participated in in-depth interviews. A deductive analysis of the interview materials was applied using Shiffman and Smith's political prioritisation framework.

Results: Several factors contributed to the underrepresentation of homeless women's health and well-being needs in the policy context. Although many governmental and non-governmental organisations contributed to the homeless-focused programme, there was little collaboration and no unifying leadership. Moreover, there was insufficient advocacy and mobilisation to pressure national leaders. Concerning ideas, there was no consensus regarding the definition of and solution to homeless women's health and social protection issues. Regarding political contexts and issue characteristics, a lack of a well-established structure, a paucity of information on the number of homeless women and the severity of their health situations relative to other problems, and the lack of clear indicators prevented this issue from gaining political priority.

Conclusions: To prioritise the health and well-being of homeless women, the government should form a unifying collaboration and a governance structure that addresses the unmet needs of these women. It is imperative to divide responsibilities and explicitly include homeless people and services targeted for them in the national health and social protection implementation documents. Further, generating consensus on framing the problems and solutions and establishing indicators for assessing the situation is vital.

Spheroid cultures of primary human hepatocytes (PHH) are used in studies of hepatic drug metabolism and toxicity. The cultures are maintained under different cone-lions, with possible confounding results. We performed an in-depth analysis of the influence of various culture conditions to find the optimal conditions for the maintenance of an in vivo like phenotype. The formation, protein expression, and function of PHH spheroids were followed for three weeks in a high-throughput 384-well format. Medium composition affected spheroid histology, global proteome profile, drug metabolism and drug-induced toxicity. No epithelial-mesenchymel transition was observed. Media with fasting glucose and insulin levels gave spheroids with phenotypes closest to normal PHH. The most expensive medium resulted in PHH features most divergent from that of native PHH. Our results provide a protocol for culture of healthy PHH with maintained function a prerequisite for studies of hepatocyte homeostasis and more reproducible hepatocyte research.

3D spheroids of primary human hepatocytes (3D PHH) retain a differentiated phenotype with largely conserved metabolic function and proteomic fingerprint over weeks in culture. As a result, 3D PHH are gaining importance as a model for mechanistic liver homeostasis studies and in vitro to in vivo extrapolation (IVIVE) in drug discovery. However, the kinetics and regulation of drug transporters have not yet been assessed in 3D PHH. Here, we used organic cation transporter 1 (OCT1/SLC22A1) as a model to study both transport kinetics and the long-term regulation of transporter activity via relevant signalling pathways. The kinetics of the OCT1 transporter was studied using the fluorescent model substrate 4-(4-(dimethylamino)styryl)-N-methylpyridinium (ASP+) and known OCT1 inhibitors in individual 3D PHH. For long-term studies, 3D PHH were treated with xenobiotics for seven days, after which protein expression and OCT1 function were assessed. Global proteomic analysis was used to track hepatic phenotypes as well as prototypical changes in other regulated proteins, such as P-glycoprotein and Cytochrome P450 3A4. ASP+ kinetics indicated a fully functional OCT1 transporter with a K_m value of 14 ± 4.0µM as the mean from three donors. Co-incubation with known OCT1 inhibitors decreased the uptake of ASP+ in the 3D PHH spheroids by 35–52%. The long-term exposure studies showed that OCT1 is relatively stable upon activation of nuclear receptor signalling or exposure to compounds that could induce inflammation, steatosis or liver injury. Our results demonstrate that 3D PHH spheroids express physiologically relevant levels of fully active OCT1 and that its transporter kinetics can be accurately studied in the 3D PHH configuration. We also confirm that OCT1 remains stable and functional during the activation of key metabolic pathways that alter the expression and function of other drug transporters and drug-metabolizing enzymes. These results will expand the range of studies that can be performed using 3D PHH.

Organotypic three-dimensional liver spheroid cultures in which hepatic cells retain their molecular phenotype and functionality have emerged as powerful tools for preclinical drug development. In recent years a multitude of culture systems have been developed; however, a thorough side-by-side benchmarking of the different methods is lacking. Here, we compared the performance of ten different 96- and 384-well microplate types to support spheroid formation and long-term culture. Specifically, we evaluated differences in spheroid formation kinetics, viability, functionality, expression patterns, and their utility for hepatotoxicity assessments using primary human hepatocytes (PHH) and primary canine hepatocytes (PCH). All 96-well plates enabled formation of PHH liver spheroids, albeit with differences between plates in spheroid size, geometry, and reproducibility. Performance of different 384-wells was less consistent. Only 6/10 microplates supported the formation of PCH aggregates. Interestingly, even if PCH aggregates in these six microplates were more loosely packed than PHH spheroids, they maintained their function and were compatible with long-term pharmacological and toxicological assays. Overall, Corning and Biofloat plates showed the best performance in the formation of both human and canine liver spheroids with highest viability, most physiologically relevant phenotypes, superior CYP activity and lowest coefficient of variation in toxicity assays. The presented data constitutes a valuable resource that demonstrates the impacts of current ultra-low attachment plates on liver spheroid metrics and can guide evidence-based plate selection. Combined, these results have important implications for the cross-comparison of different studies and can facilitate the standardization and reproducibility of three-dimensional liver culture experiments. We assessed the performance of ten different 384- and 96-well ultra-low attachment (ULA) microplates in facilitating three-dimensional spheroid culture of primary human hepatocytes (PHH) and primary canine hepatocytes (PCH) by examining effects on formation kinetics, spheroid morphology, long-term stability, functionality, expression signatures, and utility for hepatotoxicity assessment. Significant variations in experimental endpoints were observed between plates. These results can guide the optimization of spheroid experiments and contribute to the standardization of testing in three-dimensional organotypic liver cultures. image

Organotypic human tissue models constitute promising systems to facilitate drug discovery and development. They allow to maintain native cellular phenotypes and functions, which enables long-term pharmacokinetic and toxicity studies, as well as phenotypic screening. To trace relevant phenotypic changes back to specific targets or signaling pathways, comprehensive proteomic profiling is the gold-standard. A multitude of proteomic workflows have been applied on 3D tissue models to quantify their molecular phenotypes; however, their impact on analytical results and biological conclusions in this context has not been evaluated. The performance of twelve mass spectrometry-based global proteomic workflows that differed in the amount of cellular input, lysis protocols and quantification methods was compared for the analysis of primary human liver spheroids. Results differed majorly between protocols in the total number and subcellular compartment bias of identified proteins, which is particularly relevant for the reliable quantification of transporters and drug metabolizing enzymes. Using a model of metabolic dysfunction-associated steatotic liver disease, we furthermore show that critical disease pathways are robustly identified using a standardized high throughput-compatible workflow based on thermal lysis, even using only individual spheroids (1500 cells) as input. The results increase the applicability of proteomic profiling to phenotypic screens in organotypic microtissues and provide a scalable platform for deep phenotyping from limited biological material.

3D cultures of primary human hepatocytes (3D PHH) are successfully used to reduce and replace the use of animal experiments in biomedical research. Yet, the initial formation of 3D PHH is highly dependent on the supplementation with fetal bovine serum (FBS). However, the molecular composition of FBS and its effects on cultured cells are poorly understood. Moreover, FBS is prone to batch-to-batch variation, immunogenic risk and lack of adherence to the replacement, refinement, and reduction (3Rs) of animal experiments. Here, we demonstrate that FBS can be fully replaced by animal-free substitutes, thus facilitating fully chemically defined and animal serum-free 3D PHH cultures. Specifically, we combined a previously developed animal-free substitute cocktail (Rafnsdóttir et al., 2023) with a normoglycemic (5.5 mM glucose and 0.58 ng/mL insulin) chemically defined culture medium (Handin et al., 2023). Morphological and viability evaluations, along with global proteomics data, demonstrated that serum-free cultured 3D PHH have equal or superior viability and functional performance of cytochrome P450s, rendering this medium useful for long-term studies and in vitro ADMET applications. This study marks a significant advancement in the development of animal serum-free culture conditions for primary human cell cultures, paving the way for more reliable and ethical in vitro studies.

Stoner and Heisenberg excitations in magnetic materials are inherently different. The first involves an effective reduction of the exchange splitting, whereas the second comprises excitation of spin waves. In this work, we test the impact of these two excitations in the hybrid Stoner-Heisenberg system of FePd. We present a microscopic picture of ultrafast demagnetization dynamics in this alloy, which represents both components of strong local exchange splitting in Fe and induced polarization in Pd. We identify the spin-orbit coupling (SOC) and the optical intersite spin transfer (OISTR) as the two dominant factors for demagnetization at ultrashort timescales. Remarkably, the drastic difference in the origin of the magnetic moment of the Fe and Pd species is not deciding the initial magnetization dynamics in this alloy. By tuning the external laser pulse, the extrinsic OISTR can be manipulated for site-selective demagnetization on femtosecond timescales providing the fastest way for optical and selective control of the magnetization dynamics in alloys. Saliently, our results signify why various experiments demonstrating OISTR might obtain conflicting results.

The complex electronic structure of metallic ferromagnets is determined by a balance between exchange interaction, electron hopping leading to band formation, and local Coulomb repulsion. By combining high energy and temporal resolution in femtosecond time-resolved X-ray absorption spectroscopy with ab initio time-dependent density functional theory we analyze the electronic structure in fcc Ni on the time scale of these interactions in a pump-probe experiment. We distinguish transient broadening and energy shifts in the absorption spectra, which we demonstrate to be captured by electron repopulation respectively correlation-induced modifications of the electronic structure, requiring to take the local Coulomb interaction into account.

The direct manipulation of spins via light may provide a path toward ultrafast energy-efficient devices. However, distinguishing the microscopic processes that can occur during ultrafast laser excitation in magnetic alloys is challenging. Here, we study the Heusler compound Co2MnGa, a material that exhibits very strong light-induced spin transfers across the entire M-edge. By combining the element specificity of extreme ultraviolet high-harmonic probes with time-dependent density functional theory, we disentangle the competition between three ultrafast light-induced processes that occur in Co2MnGa: same-site Co-Co spin transfer, intersite Co-Mn spin transfer, and ultrafast spin flips mediated by spin-orbit coupling. By measuring the dynamic magnetic asymmetry across the entire M-edges of the two magnetic sublattices involved, we uncover the relative dominance of these processes at different probe energy regions and times during the laser pulse. Our combined approach enables a comprehensive microscopic interpretation of laser-induced magnetization dynamics on time scales shorter than 100 femtoseconds.

Ultrafast magnetization dynamics driven by ultrashort pump lasers is typically explained by changes in electronic populations and scattering pathways of excited conduction electrons. This conventional approach overlooks the fundamental role of quantum mechanical selection rules, governing transitions from core states to the conduction band, that forms the key method of the probing step in these experiments. By employing fully ab initio time-dependent density functional theory, we reveal that these selection rules profoundly influence the interpretation of ultrafast spin dynamics at specific probe energies. Our analysis for hcp Co and fcc Ni at the M edge demonstrates that the transient dynamics, as revealed in pump-probe experiments, arise from a complex interplay of optical excitations of the M shell. Taking into account the selection rules and conduction electron spin flips, this leads to highly energy-dependent dynamics. These findings address longstanding discrepancies in experimental TMOKE measurements and show that only through meticulous consideration of matrix elements at the probe stage, can one ensure that magnetization dynamics is revealed in its true nature, instead of being muddled by artifacts arising from the choice of probe energy.

The interplay between various degrees of freedom in laser induced ultrafast magnetization dynamics (LIUMD) of magnetic alloys is intricate due to the competition between different mechanisms and processes. In this work, we resolve the element specific magnetization dynamics of FePd alloy and further elucidate the dependency of the OISTR mechanism on the laser pulse parameters using ultrashort, short and relatively longer pulse duration with weak and strong fluence. Remarkably, our results illustrate potential discrepancies in experiments measuring the optical inter site spin transfer (OISTR) effect in magnetic alloys.

In natural photosynthesis, the light-driven splitting of water into electrons, protons and molecular oxygen forms the first step of the solar-to-chemical energy conversion process. The reaction takes place in photosystem II, where the Mn₄CaO₅ cluster first stores four oxidizing equivalents, the S₀ to S₄ intermediate states in the Kok cycle, sequentially generated by photochemical charge separations in the reaction center and then catalyzes the O–O bond formation chemistry^1,2,3. Here, we report room temperature snapshots by serial femtosecond X-ray crystallography to provide structural insights into the final reaction step of Kok’s photosynthetic water oxidation cycle, the S₃→[S₄]→S₀ transition where O₂ is formed and Kok’s water oxidation clock is reset. Our data reveal a complex sequence of events, which occur over micro- to milliseconds, comprising changes at the Mn₄CaO₅ cluster, its ligands and water pathways as well as controlled proton release through the hydrogen-bonding network of the Cl1 channel. Importantly, the extra O atom O_x, which was introduced as a bridging ligand between Ca and Mn1 during the S₂→S₃ transition^4,5,6, disappears or relocates in parallel with Y_z reduction starting at approximately 700 μs after the third flash. The onset of O₂ evolution, as indicated by the shortening of the Mn1–Mn4 distance, occurs at around 1,200 μs, signifying the presence of a reduced intermediate, possibly a bound peroxide.

Photosystem II starts the photosynthetic electron transport chain that converts solar energy into chemical energy and thus sustains life on Earth. It catalyzes two chemical reactions: water oxidation to molecular oxygen and plastoquinone reduction. Coupling of electron and proton transfer is crucial for efficiency; however, the molecular basis of these processes remains speculative owing to uncertain water binding sites and the lack of experimentally determined hydrogen positions. We thus collected high-resolution cryo-electron microscopy data of fully hydrated photosystem II from the thermophilic cyanobacterium Thermosynechococcus vestitus to a final resolution of 1.71 angstroms. The structure reveals several previously undetected partially occupied water binding sites and more than half of the hydrogen and proton positions. This clarifies the pathways of substrate water binding and plastoquinone B protonation.

Light-driven water oxidation by photosystem II sustains life on Earth by providing the electrons and protons for the reduction of CO2 to carbohydrates and the molecular oxygen we breathe. The inorganic core of the oxygen evolving complex is made of the earth-abundant elements manganese, calcium and oxygen (Mn₄CaO₅ cluster), and is situated in a binding pocket that is connected to the aqueous surrounding via water-filled channels that allow water intake and proton egress. Recent serial crystallography and infrared spectroscopy studies performed with PSII isolated from Thermosynechococcus vestitus (T. vestitus) support that one of these channels, the O1 channel, facilitates water access to the Mn₄CaO₅ cluster during its S₂ - S₃ and S₃ - S₄ - S₀ state transitions, while a subsequent CryoEM study concluded that this channel is blocked in the cyanobacterium Synechocystis sp. PCC 6803, questioning the role of the O1 channel in water delivery. Employing site-directed mutagenesis we modified the two O1 channel bottleneck residues D1-E329 and CP43-V410 (T. vestitus numbering) and probed water access and substrate exchange via time resolved membrane inlet mass spectrometry. Our data demonstrates that water reaches the Mn₄CaO₅ cluster via the O1 channel in both wildtype and mutant PSII. In addition, the detailed analysis provides functional insight into the intricate protein-water-cofactor network near the Mn₄CaO₅ cluster that includes the pentameric, near planar ‘water wheel’ of the O1 channel.  

Water oxidation by photosystem II (PSII) sustains most life on Earth, but the molecular mechanism of this unique process remains controversial. The ongoing identification of the binding sites and modes of the two water-derived substrate oxygens (‘substrate waters’) in the various intermediates (Si states, i = 0, 1, 2, 3, 4) that the water-splitting tetra-manganese calcium penta-oxygen (Mn4CaO5) cluster attains during the reaction cycle provides central information towards resolving the unique chemistry of biological water oxidation. Mass spectrometric measurements of single- and double-labeled dioxygen species after various incubation times of PSII with H218O provide insight into the substrate binding modes and sites via determination of exchange rates. Such experiments have revealed that the two substrate waters exchange with different rates that vary independently with the Si state and are hence referred to as the fast (Wf) and the slow (WS) substrate waters. New insight for the molecular interpretation of these rates arises from our recent finding that in the S2 state, under special experimental conditions, two different rates of WS exchange are observed that appear to correlate with the high spin and low spin conformations of the Mn4CaO5 cluster. Here, we reexamine and unite various proposed methods for extracting and assigning rate constants from this recent data set. The analysis results in a molecular model for substrate-water binding and exchange that reconciles the expected non-exchangeability of the central oxo bridge O5 when located between two Mn(IV) ions with the experimental and theoretical assignment of O5 as WS in all S states. The analysis also excludes other published proposals for explaining the water exchange kinetics.

Photosystem II (PSII) catalyzes the water-splitting reaction during photosynthesis, generating oxygen, protons, and electrons through the multi-step S state cycle. The role of protein residues in maintaining the intricate dynamics of substrate water exchange within the Mn4CaO5 cluster remains a critical area of research. In this study, we investigate the effects of the D1-N298A mutation on water exchange kinetics in PSII core complexes. Comparing wild-type (WT) PSII to the mutant, we observed that the mutation significantly slows the exchange of both substrate waters in the S3-state, while leaving the slow S2 state exchange kinetics largely unaffected. Our results suggest that the D1-N298 residue plays a vital role in maintaining the hydrogen-bond network within the O1 channel, which facilitates both proton-coupled electron transfer (PCET) during S state turnover and substrate water exchange. The disruption of the YZ-H190-N298 triad by the D1-N298A mutation is proposed to alter the structural organization of the waterwheel, raising the activation energy for Mn1 reduction by YZ and impairing protonation and deprotonation processes. These findings reinforce the hypothesis that water molecules introduced via the O1 channel, such as Wx or W3, act as the fast-exchanging substrate in the S2 state and become Ox/O6 in the S3 state, forming the O–O bond with the slowly-exchanging substrate identified previously as the central µ3-oxo bridge O5.

Severe infections are life-threatening conditions commonly seen in the intensive care units (ICUs). Antibiotic treatment with adequate concentrations is of great importance during the first days when the bacterial load is the highest. Therapeutic drug monitoring (TDM) of β-lactam antibiotics has been suggested to monitor target attainment and to improve the outcome. This prospective multi-center study in seven ICUs in Sweden investigated pharmacokinetic/pharmacodynamic-target (PK/PD-target) attainment for cefotaxime, piperacillin-tazobactam and meropenem, commonly used β-lactams in Sweden. A mid-dose and trough antibiotic concentration blood sample were taken from patients with severe infection daily during the first 72 h of treatment. Antibiotic plasma concentrations were analysed by liquid chromatography-mass spectrometry (LC–MS). Antibiotic concentrations 100% time above MIC (minimal inhibitory concentration), (100% T > MIC) and four times above MIC 50% of the time (50% T > 4xMIC) were used as PK/PD-targets. We included 138 patients with the median age of 67 years and the median Simplified Acute Physiology Score 3 (SAPS3) of 59. Forty-five percent of the study-population failed to reach 100% T > MIC during the first day of treatment. The results were similar the following two days. There was a three-fold risk of not meeting the PK/PD target if the patient was treated with cefotaxime. For the cefotaxime treated patients 8 out of 55 (15%) had at least one end-dose concentrations below the level of detection during the study. Low age, low illness severity, low plasma creatinine, lower respiratory tract infection and cefotaxime treatment were risk factors for not reaching 100% T > MIC. In Swedish ICU-patients treated with β-lactam antibiotics, a high proportion of patients did not reach the PK/PD target. TDM could identify patients that need individual higher dosing regimens already on the first day of treatment. Further studies on optimal empirical start dosing of β-lactams, especially for cefotaxime, in the ICU are needed.

Background

Therapeutic drug monitoring (TDM) has been suggested to optimize antimicrobial target attainment, typically using 100%T_>MIC, in β-lactam treatment in the ICU. The MIC parameter used in this equation is mostly the worst case scenario MIC (MIC_WCS)—the highest MIC the empirical treatment should cover. However, the impact of the MIC parameter used in pharmacokinetic/pharmacodynamic calculations has been poorly investigated.

Objectives

To assess the influence of target attainment rates for two different MIC parameters using actual MICs of the causative pathogens as the primary reference.

Methods

In a Swedish multicentre study of target attainment for 138 ICU patients treated with β-lactams, the causative pathogen was isolated and subjected to reference MIC testing. Whenever the strain belonged to the WT distribution, we assigned it to the category MIC_ECOFF (epidemiological cut-off value). In the calculations we compared the MIC_ECOFF and the MIC_WCS.

Results

The proportion of patients with target attainment failure for all antibiotics using 100%T_>MIC was 45% (95% CI, 37%–53%) for MIC_WCS and 23% (95% CI, 16%–31%) for MIC_ECOFF. When the target 50%T_>4×MIC was used, corresponding attainment failures were 57% (95% CI, 49%–66%) and 25% (95% CI, 17%–32%) for MIC_WCS and MIC_ECOFF, respectively.

Conclusions

MIC_WCS can overestimate target attainment failure. The use of MIC_WCS could be one reason for the difficulties in establishing a relationship between target failure and mortality in other studies. Based on findings herein, the MIC_ECOFF, which is based on the MIC of the causative pathogen, should be considered a more suitable alternative. When no pathogen is detected, the MIC_ECOFF of likely pathogens according to infection type should be used.

OBJECTIVES: This study aimed to predict the impact of different infusion strategies on pharmacokinetic/pharmacodynamic (PK/PD) target attainment and the potential risk for toxicity in an ICU cohort treated with β-lactams.

METHOD: Using collected patient data from 137 adult ICU patients, and applying population PK models, individual PK parameters were estimated and used to predict concentrations and target attainment following cefotaxime 2 g q8h, piperacillin/tazobactam 4.5 g q6h and meropenem 1 g q8h, applying 15 min short infusions (SI), 3 h extended infusions (EI) and 24 h continuous infusion (CI). The MIC level of the most common primary pathogens, and the worst-case scenario (WCS) pathogen, were used in analyses.

RESULTS: For primary pathogens, target was reached in 94% (129/137) using SI. For WCS pathogens treated with piperacillin/tazobactam and meropenem, 78% (65/83) and 92% (76/83) reached target using SI and EI, respectively. However, target attainment was lower for cefotaxime [SI: 31% (17/54), EI: 44% (24/54)]. Overall, the number of individuals with potentially toxic concentrations was low, both in EI (n = 7) and SI (n = 5). For CI and WCS, target was reached in 50% (27/54), 96% (54/56) and 93% (25/27) for cefotaxime, piperacillin/tazobactam and meropenem, respectively.

CONCLUSIONS: In a Swedish ICU cohort target attainment rates for primary pathogens were high regardless of infusion strategy. In WCS pathogens, SI was insufficient, suggesting the benefit of routine use of EI or CI. However, for cefotaxime, target attainment remained low also with EI and CI. The use of CI might lead to unnecessarily high concentrations, but well-established toxicity levels are lacking and future studies are warranted.

Escherichia coli rRNAs are post-transcriptionally modified at 36 positions but their modification enzymes are dispensable individually for growth, bringing into question their significance. However, a major growth defect was reported for deletion of the RlmE enzyme, which abolished a 2 ' O methylation near the peptidyl transferase center (PTC) of the 23S rRNA. Additionally, an adjacent 80-nt "critical region" around the PTC had to be modified to yield significant peptidyl transferase activity in vitro. Surprisingly, we discovered that an absence of just two rRNA modification enzymes is conditionally lethal (at 20 degrees C): RlmE and RluC. At a permissive temperature (37 degrees C), this double knockout was shown to abolish four modifications and be defective in ribosome assembly, though not more so than the RlmE single knockout. However, the double knockout exhibited an even lower rate of tripeptide synthesis than did the single knockout, suggesting an even more defective ribosomal translocation. A combination knockout of the five critical-region-modifying enzymes RluC, RlmKL, RlmN, RlmM, and RluE (not RlmE), which synthesize five of the seven critical-region modifications and 14 rRNA and tRNA modifications altogether, was viable (minor growth defect at 37 degrees C, major at 20 degrees C). This was surprising based on prior in vitro studies. This five-knockout combination had minimal effects on ribosome assembly and frameshifting at 37 degrees C, but greater effects on ribosome assembly and in vitro peptidyl transferase activity at cooler temperatures. These results establish the conditional essentiality of bacterial rRNA modification enzymes and also reveal unexpected plasticity of modification of the PTC region in vivo.

Large ribosomal RNAs (rRNAs) are modified heavily post-transcriptionally in functionally important regions but, paradoxically, individual knockouts (KOs) of the modification enzymes have minimal impact on Escherichia coli growth. Furthermore, we recently constructed a strain with combined KOs of five modification enzymes (RluC, RlmKL, RlmN, RlmM and RluE) of the ‘critical region’ of the peptidyl transferase centre (PTC) in 23S rRNA that exhibited only a minor growth defect at 37°C (although major at 20°C). However, our combined KO of modification enzymes RluC and RlmE (not RluE) resulted in conditional lethality (at 20°C). Although the growth rates for both multiple-KO strains were characterized, the molecular explanations for such deficits remain unclear. Here, we pinpoint biochemical defects in these strains. In vitro fast kinetics at 20°C and 37°C with ribosomes purified from both strains revealed, counterintuitively, the slowing of translocation, not peptide bond formation or peptidyl release. Elongation rates of protein synthesis in vivo, as judged by the kinetics of β-galactosidase induction, were also slowed. For the five-KO strain, the biggest deficit at 37°C was in 70S ribosome assembly, as judged by a dominant 50S peak in ribosome sucrose gradient profiles at 5 mM Mg²⁺. Reconstitution of this 50S subunit from purified five-KO rRNA and ribosomal proteins supported a direct role in ribosome biogenesis of the PTC region modifications per se, rather than of the modification enzymes. These results clarify the importance and roles of the enigmatic rRNA modifications.

Translation terminates by releasing the polypeptide chain in one of two chemical reactions catalyzed by the ribosome. Release is also a target for engineering, as readthrough of a stop codon enables incorporation of unnatural amino acids and treatment of genetic diseases. Hydrolysis of the ester bond of peptidyl-tRNA requires conformational changes of both a class I release factor (RF) protein and the peptidyl transferase center of a large subunit rRNA. The rate-limiting step was proposed to be hydrolysis at physiological pH and an RF conformational change at higher pH, but evidence was indirect. Here, we tested this by activating the ester electrophile at the Escherichia coli ribosomal P site using a trifluorine-substituted amino acid. Quench-flow kinetics revealed that RF1-catalyzed release could be accelerated, but only at pH 6.2-7.7 and not higher pH. This provided direct evidence for rate-limiting hydrolysis at physiological or lower pH and a different rate limitation at higher pH. Additionally, we optimized RF-free release catalyzed by unacylated tRNA or the CCA trinucleotide (in 30% acetone). We determined that these two model release reactions, although very slow, were surprisingly accelerated by the trifluorine analog but to a different extent from each other and from RF-catalyzed release. Hence, hydrolysis was rate limiting in all three reactions. Furthermore, in 20% ethanol, we found that there was significant competition between fMet-ethyl ester formation and release in all three release reactions. We thus favor proposed mechanisms for translation termination that do not require a fully-negatively-charged OH⁻ nucleophile.

In this work, high-performance four-terminal solution-processed tandem solar cells were fabricated by using dye-sensitized solar cells (DSSCs) as top-cells and lead sulfide (PbS) colloidal quantum dot solar cells (CQDSCs) as bottom-cells. For dye-sensitized top-cells, three different dye combinations were used while the titanium dioxide (TiO2) scattering layer was removed to maximize the transmission. For the PbS bottom-cells, quantum dots with different sizes were compared. Over 12% power conversion efficiency has been achieved by using the XL dye mixture and 890 nm PbS QDs, which shows a significant efficiency enhancement when compared to single DSSC or CQDSC subcells.

In this work, four-terminal (4T) tandem solar cells were fabricated by using a methylammonium lead iodide (MAPbI₃) perovskite solar cell (PSC) as the front-cell and a lead sulfide (PbS) colloidal quantum dot solar cell (CQDSC) as the back-cell. Different modifications of the tandem interlayer, at the interface between the sub-cells, were tested in order to improve the infrared transparency of the perovskite sub-cell and consequently increase the utilization of infrared (IR) light by the tandem system. This included the incorporation of a semi-transparent thin gold electrode (Au) on the MAPbI₃ solar cell, followed by adding a molybdenum(VI) oxide (MoO₃) layer or a surlyn layer. These interlayer modifications resulted in an increase of the IR transmittance to the back cell and improved the optical stability, compared to that in the reference devices. This investigation shows the importance of the interlayer, connecting the PSC with a strong absorption in the visible region and the CQDSC with a strong infrared absorption to obtain efficient next-generation tandem photovoltaics (PVs).

Renewable energy sources, such as wind and solar power, are increasingly important today to reduce emissions from fossil-based energy sources. However, the electricity from wind and solar power varies over time and depends on weather conditions and the time of the day. Therefore, to include a large fraction of electricity from these energy sources in the electricity grid, large-scale and low-cost energy storage is needed. Herein, it is investigated how a combination of quantum dot based photovoltaic cells and perovskite-based photovoltaic cells can be used to increase the energy conversion efficiency and increase the working range of energy storage devices based on conversion between heat, light, and electricity. The results show that these new types of photovoltaic materials have very promising properties for efficient utilization in energy storage devices, which have the potential for large-scale and low-cost energy storage.

The p-type semiconducting colloidal quantum dot (CQD), working as a hole conductor in CQD solar cells (CQDSCs), is critical for charge carrier extraction and therefore, to large extent, determines the device's photovoltaic performance. However, during the preparation of a p-type CQD solid film on the top of an n-type CQD solid film, forming a p-n heterojunction within the CQDSCs, the optoelectronic properties of the underlayered n-type CQD solid film are significantly affected by conventional 1,2-ethanedithiol (EDT) ligands due to its high reactivity. Herein, a series of thiol ligands are comprehensively studied for p-type CQDs, which suggests that, by finely controlling the interaction between the CQDs and thiol ligands during the preparation of p-type CQD solid films, the n-type CQD solid films can be well protected and avoid destruction induced by thiol ligands. The p-type CQD solid film with 4-aminobenzenethiol (ABT) passivating the CQD surface exhibits better optoelectronic properties than the conventional p-type EDT-based CQD solid films, resulting in an improved photovoltaic performance in CQDSCs.

Time-resolved photoelectron spectroscopy can give insights into carrier dynamics and offers the possibility of element and site-specific information through the measurements of core levels. In this paper, we demonstrate that this method can access electrons dynamics in PbS quantum dots over a wide time window spanning from pico- to microseconds in a single experiment carried out at the synchrotron facility BESSY II. The method is sensitive to small changes in core level positions. Fast measurements at low pump fluences are enabled by the use of a pump laser at a lower repetition frequency than the repetition frequency of the X-ray pulses used to probe the core level electrons: Through the use of a time-resolved spectrometer, time-dependent analysis of data from all synchrotron pulses is possible. Furthermore, by picosecond control of the pump laser arrival at the sample relative to the X-ray pulses, a time-resolution limited only by the length of the X-ray pulses is achieved. Using this method, we studied the charge dynamics in thin film samples of PbS quantum dots on n-type MgZnO substrates through time-resolved measurements of the Pb 5d core level. We found a time-resolved core level shift, which we could assign to electron injection and charge accumulation at the MgZnO/PbS quantum dots interface. This assignment was confirmed through the measurement of PbS films with different thicknesses. Our results therefore give insight into the magnitude of the photovoltage generated specifically at the MgZnO/PbS interface and into the timescale of charge transport and electron injection, as well as into the timescale of charge recombination at this interface. It is a unique feature of our method that the timescale of both these processes can be accessed in a single experiment and investigated for a specific interface.

Charge transfer is essential for all electrochemical processes, such as in batteries where it is facilitated through the incorporation of ion–electron pairs into solid crystals. The low solubility of lithium (Li) in some of these host lattices cause phase changes, which for example happens in FePO₄. This results in the growth of interfacial patterns at the mesoscale between a Li-poor and Li-rich phase, FePO₄ and LiFePO₄ respectively. Conventionally, the effect of charge transfer on the evolution of these phases is usually modelled using the Butler–Volmer equation. However, the exponentially increasing current–overpotential relation in this formalism becomes problematic for battery systems operating under high currents. In this study, we implement a phase-field model to investigate two electrochemical reaction models: the Butler–Volmer and the Marcus–Hush–Chidsey formulation. We assess their effect on the spatial and temporal evolution of the FePO₄ and LiFePO₄ phases. Both reaction models demonstrate similar microstructural patterns in equilibrium. Nevertheless, a significant increase in current density is caused by using the Butler–Volmer expression, leading to an accelerated reaction rate at high overpotentials and an exaggerated delithiation. Furthermore, we show that including anisotropic elastic strain fields in the phase-field model accelerates the delithiation process, reaching the bulk mass transport limitation faster. These elastic effects, when included in the overpotential, can cause the current density to exceed its limits, a problem inherently mitigated by the Marcus–Hush–Chidsey model.

Energy is stored in a LiFePO₄ battery electrode through the intercalation of Li. As Li incorporate into the crystal lattice of Fe⁡(III)⁢PO₄, electrons reduce Fe(III) into Fe(II). The interactions of Li and its vacant site (Va) with these localized electrons (holes), so-called polarons, cause phase separation during battery operation. These fundamental interactions are however difficult to quantify using standard electronic structure calculations. In this paper, we utilize DFT+𝑈 with occupation matrix control to compute interaction energies at varying Li-Fe(II) and Va-Fe(III) pair separations. The increased energy with separation warrants the use of an electrostatic description. The DFT+𝑈 data are fitted to a Coulombic potential with two-body corrections and used in a Monte Carlo scheme. The coordination of the species determines their short-range ordering, showing that the Li and Va create chains bridged by their associated polarons which dissociate into dimers at higher temperatures. This dissociation happens at higher temperatures for Va than for Li, indicating a more pronounced clustering behavior during the formation of FePO₄. Notably, a significant amount of uncoordinated Li exists at elevated temperatures, challenging the simplified picture of complete Li-Fe(II) pairing.

Background: Carbapenem-resistant bacteria pose a threat to public health. Characterising thepharmacokinetics-pharmacodynamics (PKPD) of meropenem longitudinally in vivo against resistant bacteria could provide valuable information for development and translation of carbapenem-based therapies.

Objectives: To assess the time course of meropenem effects in vivo against strains with high MIC topredict PK/PD indices and expected efficacy in patients using a modelling approach.

Methods: A PKPD model was built on longitudinal bacterial count data to describe meropenem effectsagainst six Escherichia coli and Klebsiella pneumoniae strains (MIC values 32–128 mg/L) in a 24 h mousethigh infection model. The model was used to derive PK/PD indices from simulated studies in mice andto predict the efficacy of different infusion durations with high-dose meropenem (2 g q8 h/q12 h fornormal/reduced kidney function) in patients.

Results: Data from 592 mice were available for model development. The estimated meropenemconcentration-dependent killing rate was not associated with differences in MIC. The fraction of timethat unbound concentrations exceeded EC50 (fT>EC50, EC50 = 1.01 mg/L) showed higher correlations thanfT>MIC. For all investigated strains, bacteriostasis at 24 h was predicted for prolonged infusions of highdose meropenem monotherapy in >90% of patients.

Conclusions: The developed PKPD model successfully described bacterial growth and meropenem killingover time in the thigh infection model. For the investigated strains, the MIC, determined in vitro, orMIC-based PK/PD indices, did not predict in vivo response. Simulations suggested prolonged infusions ofhigh-dose meropenem to be efficacious in patients infected by the studied strains

Background: Translation of experimental data on antibiotic activity typically relies on pharmacokinetic/pharmacodynamic (PK/PD) indices. Model-based approaches, considering the full antibiotic killing time course, could be an alternative.

Objectives: To develop a mechanism-based modelling framework to assess the in vitro and in vivo activity of the FabI inhibitor antibiotic afabicin, and explore the ability of a model built on in vitro data to predict in vivo outcome.

Methods: A PK/PD model was built to describe bacterial counts from 162 static in vitro time-kill curves evaluating the effect of afabicin desphosphono, the active moiety of the prodrug afabicin, against 21 Staphylococcus aureus strains. Combined with a mouse PK model, outcomes of afabicin doses of 0.011-190 mg/kg q6h against nine S. aureus strains in a murine thigh infection model were predicted, and thereafter refined by estimating PD parameters.

Results: A sigmoid Emax model, with EC50 scaled by the MIC described the afabicin desphosphono killing in vitro. This model predicted, without parameter re-estimation, the in vivo bacterial counts at 24 h within a ±1 log margin for most dosing groups. When parameters were allowed to be estimated, EC50 was 38%-45% lower in vivo, compared with in vitro, within the studied MIC range.

Conclusions: The developed PK/PD model described the time course of afabicin activity across experimental conditions and bacterial strains. This model showed translational capacity as parameters estimated on in vitro time-kill data could well predict the in vivo outcome for a wide variety of doses in a mouse thigh infection model.

In intensive care, drugs are commonly administered through central venous catheters (CVC). These catheters and central venous dialysis catheters (CVDC) are often placed in the same vessel for practical reasons. The aim of this experimental study was to investigate if the position of CVC and CVDC influences the elimination of infused drugs, during continuous renal replacement therapy (CRRT). In a randomized, cross-over model, anesthetized piglets received both a CVC and a CVDC in a jugular vein. Another CVDC was placed in a femoral vein for comparison. After baseline measurements, CRRT was performed in either of the CVDC, each CRRT-period separated by another baseline period. Hypotension was induced by peripherally given sodium nitroprusside. In the CVC, both gentamicin and noradrenaline were administered. Noradrenaline was titrated to reach a target blood pressure. When CRRT was performed using the CVDC in the same vessel as the drugs were infused, the plasma concentration of gentamicin was reduced compared with when the infusion and CVDC were in different vessels (5.66 [standard deviation (SD) ± 1.23] vs. 7.76 [SD ± 2.30] mg/L [p = 0.02]). The noradrenaline infusion rate needed to reach the target blood pressure was more than doubled (0.32 [SD ± 0.16] vs. 0.15 [SD ± 0.08] µg/kg/min [p = 0.006]). This experimental study indicates that the removal of drugs is increased if infusion is in close vicinity of the CVDC, during CRRT.

BACKGROUND: The aim of this experimental study was to elucidate whether different distances between central venous catheter tips can affect drug clearance during continuous renal replacement therapy (CRRT). Central venous catheters (CVCs) are widely used in intensive care patients for drug infusion. If a patient receives CRRT, a second central dialysis catheter (CDC) is required. Where to insert CVCs is directed by guidelines, but recommendations regarding how to place multiple catheters are scarce. There are indications that a drug infused in a CVC with the tip close to the tip of the CDC, could be directly aspirated into the dialysis machine, with a risk of increased clearance. However, studies on whether clearance is affected by different CVC and CDC tip positions, when the two catheters are in the same vessel, are few.

METHODS: In this model with 18 piglets, gentamicin (GM) and vancomycin (VM) were infused through a CVC during CRRT. The CVC tip was placed in different positions in relation to the CDC tip from caudal, i.e., proximal to the heart, to cranial, i.e., distal to the heart. Serum and dialysate concentrations were sampled after approximately 30 min of CRRT at four different positions: when the CVC tip was 2 cm caudally (+ 2), at the same level (0), and at 2 (- 2) and 4 (- 4) cm cranially of the tip of the CDC. Clearance was calculated. A mixed linear model was performed, and level of significance was set to p < 0.05.

RESULTS: Clearance of GM had median values at + 2 cm, 0 cm, - 2 cm and - 4 cm of 17.3 (5.2), 18.6 (7.4), 20.0 (16.2) and 26.2 (12.2) ml/min, respectively (p = 0.04). Clearance of VM had median values at + 2 cm, 0 cm, - 2 cm and - 4 cm of 16.2 (4.5), 14.7 (4.9), 19.0 (10.2) and 21.2 (11.4) ml/min, respectively (p = 0.02).

CONCLUSIONS: The distance between CVC and CDC tips can affect drug clearance during CRRT. A cranial versus a caudal tip position of the CVC in relation to the tip of the CDC led to the highest clearance.

BACKGROUND: In intensive care, different central venous catheters (CVC) are often used for infusion of drugs. If a patient is treated with continuous renal replacement therapy (CRRT) a second catheter, a central venous dialysis catheter (CVDC), is needed. Placing the catheters close together might pose a risk that a drug infused in a CVC could be directly aspirated into a CRRT machine and cleared from the blood without giving the effect intended. The purpose of this study was to elucidate if drug clearance is affected by different catheter placement, during CRRT. In this endotoxaemic animal model, an infusion of antibiotics was administered in a CVC placed in the external jugular vein (EJV). Antibiotic clearance was compared, whether CRRT was through a CVDC placed in the same EJV, or in a femoral vein (FV). To reach a target mean arterial pressure (MAP), noradrenaline was infused through the CVC and the dose was compared between the CDVDs.

RESULTS: The main finding in this study was that clearance of antibiotics was higher when both catheter tips were in the EJV, close together, compared to in different vessels, during CRRT. The clearance of gentamicin was 21.0 ± 7.3 vs 15.5 ± 4.2 mL/min (p 0.006) and vancomycin 19.3 ± 4.9 vs 15.8 ± 7.1 mL/min (p 0.021). The noradrenaline dose to maintain a target MAP also showed greater variance with both catheters in the EJV, compared to when catheters were placed in different vessels.

CONCLUSION: The results in this study indicate that close placement of central venous catheter tips could lead to unreliable drug concentration, due to direct aspiration, during CRRT.

In this work, we have studied how the choice of the continuous phase oils affects droplet-internal acoustic manipulation and mapped the acoustic properties of the selected oils to evaluate their compatibility with droplet acoustofluidic methods. The selection of continuous phase included hydrocarbon, fluorinated and silicone oils. To map the oils’ acoustic properties, we measured their speed of sound and density. We then studied the acoustic performance of each oil for droplet-internal manipulation through experiments and finite-element simulations (COMSOL Multiphysics®).From our results, we conclude that a match between the speed of sound of the continuous and dispersed phases is strongly correlated to the generation of a strong and uniform acoustic field inside the droplet. We demonstrate that conventionally-favoured fluorinated oils in droplet microfluidics are no longer the best choice when also considering droplet-internal acoustic focusing. Instead, hydrocarbon oils, especially linseed oil, are most suitable for this specific application as they generate stable and monodisperse droplets and bear the most resemblance to water in terms of acoustic properties. We believe this collection of data will serve the acoustofluidics community by providing results that aid in the selection of continuous phase in future droplet acoustofluidic studies and data for performing acoustofluidic simulations.

Droplet microfluidics has emerged as a powerful tool for miniaturizing biological assays. The continuous phase in droplet microfluidics often consists of fluorinated oils and fluorosurfactants. However, these fluids belong to a group of chemicals called PFASs (per-and polyfluoroalkyl substances), which are hazardous for the environment as they are not broken down but remain in the surrounding for a long time. As a result, PFAS chemicals are likely to be unavailable on the market in the near future. Consequently, there is a need to find replacements for fluorinated oils and fluorosurfactants that work well for droplet microfluidics. With this article, we aim to bring awareness to the issue and begin by reviewing the problem with using fluorinated chemicals in droplet microfluidics. Next, we compare the performance of four different oil/surfactant systems: two containing fluorinated oil and two without, to present alternatives for the droplet microfluidics community. Our results show that suitable alternative oil/surfactant systems exist that can effectively replace the conventional fluorinated solutions in droplet microfluidics.

The generation of hydrogel droplets using droplet microfluidics has emerged as a powerful tool with many applications in biology and medicine. Here, a microfluidic system to control the position of particles (beads or astrocyte cells) in hydrogel droplets using bulk acoustic standing waves is presented. The chip consisted of a droplet generator and a 380 µm wide acoustic focusing channel. Droplets comprising hydrogel precursor solution (polyethylene glycol tetraacrylate or a combination of polyethylene glycol tetraacrylate and gelatine methacrylate), photoinitiator and particles were generated. The droplets passed along the acoustic focusing channel where a half wavelength acoustic standing wave field was generated, and the particles were focused to the centre line of the droplets (i.e. the pressure nodal line) by the acoustic force. The droplets were cross-linked by exposure to UV-light, freezing the particles in their positions. With the acoustics applied, 89 ± 19% of the particles (polystyrene beads, 10 µm diameter) were positioned in an area ± 10% from the centre line. As proof-of-principle for biological particles, astrocytes were focused in hydrogel droplets using the same principle. The viability of the astrocytes after 7 days in culture was 72 ± 22% when exposed to the acoustic focusing compared with 70 ± 19% for samples not exposed to the acoustic focusing. This technology provides a platform to control the spatial position of bioparticles in hydrogel droplets, and opens up for the generation of more complex biological hydrogel structures.

Hydrogels are widely used for cell encapsulation and culture due to their ability to mimic extracellular matrix. Cell-laden hydrogels in the micrometer scale — microgels — offer advantages for single-cell analysis and scaffold-based 3D cell culture. However, challenges such as off-center encapsulation and asymmetric biochemical exposure lead to cellular escape and heterogeneity in cell assemblies, which compromise long-term culture and affects experimental reproducibility. Addressing these limitations requires precise three-dimensional (3D) spatial control of cells within microgels. This study presents 3D acoustic focusing of polystyrene microbeads within gelatin methacyraloyl (GelMA) hydrogel, achieved in a microwell-based acoustofluidic system that employs dual piezoelectric transducers to generate half-wavelength resonances across all three dimensions of cuboid-shaped microwells. Our findings demonstrate a versatile approach for active 3D centering of cells and microparticles in microgels, demonstrating the potential of bulk acoustic wave technologies as a biofabrication technique for biomedical research and regenerative medicine.

Background: Superparamagnetic iron oxide nanoparticles (SPIO) have been used as a tracer for sentinel lymph node (SLN) localization in breast cancer, demonstrating comparable performance to the combination of radioisotope (RI) and blue dye (BD).

Methods: A systematic literature search and meta-analysis with subgroup and meta-regression analysis were undertaken to update the available evidence, assess technique evolution, and define knowledge gaps. Recommendations were made using the GRADE approach.

Results: In 20 comparative studies, the detection rate was 97.5 per cent for SPIO and 96.5 per cent for RI +/- BD (risk ratio 1.006, 95 per cent c.i. 0.992 to 1.019; P = 0.376, high-certainty evidence). Neoadjuvant therapy, injection site, injection volume or nodal metastasis burden did not affect the detection rate, but injection over 24 h before surgery increased the detection rate on meta-regression. Concordance was 99.0 per cent and reverse concordance 97.1 per cent (rate difference 0.003, 95 per cent c.i. -0.009 to 0.015; P = 0.656, high-certainty evidence). Use of SPIO led to retrieval of slightly more SLNs (pooled mean 1.96 versus 1.89) with a higher nodal detection rate (94.1 versus 83.5 per cent; RR 1.098, 1.058 to 1.140; P < 0.001; low-certainty evidence). In meta-regression, injection over 24 h before surgery increased the SPIO nodal yield over that of RI +/- BD. The skin-staining rate was 30.8 per cent (very low-certainty evidence), and possibly prevented with use of smaller doses and peritumoral injection.

Conclusion: The performance of SPIO is comparable to that of RI +/- BD. Preoperative injection increases the detection rate and nodal yield, without affecting concordance. Whether skin staining and MRI artefacts are reduced by lower dose and peritumoral injection needs to be investigated.

Importance  Guidewires have been the standard for breast lesion localization but pose operative and logistic challenges. Paramagnetic seeds have shown promising results, but to the authors’ knowledge, no randomized comparison has been performed.

Objective  To determine whether the combination of a paramagnetic seed and superparamagnetic iron oxide (SPIO) is equivalent to guidewire and SPIO for breast cancer localization and sentinel lymph node detection (SLND).

Design, Setting, and Participants  This was a phase 3, pragmatic, equivalence, 2-arm, open-label, randomized clinical trial conducted at 3 university and/or community hospitals in Sweden from May 2018 to May 2022. Included in the study were patients with early breast cancer planned for breast conservation and SLND. Study data were analyzed July to November 2022.

Interventions  Participants were randomly assigned 1:1 to a paramagnetic seed or a guidewire. All patients underwent SLND with SPIO.

Main Outcomes and Measures  Re-excision rate and resection ratio (defined as actual resection volume / optimal resection volume).

Results  A total of 426 women (median [IQR] age, 65 [56-71] years; median [IQR] tumor size, 11 [8-15] mm) were included in the study. The re-excision rate was 2.90% (95% CI, 1.60%-4.80%), and the median (IQR) resection ratio was 1.96 (1.15-3.44). No differences were found between the guidewire and the seed in re-excisions (6 of 211 [2.84%] vs 6 of 209 [2.87%]; difference, −0.03%; 95% CI, −3.20% to 3.20%; P = .99) or resection ratio (median, 1.93; IQR, 1.18-3.43 vs median, 2.01; IQR, 1.11-3.47; P = .70). Overall SLN detection was 98.6% (95% CI, 97.1%-99.4%) with no differences between arms (203 of 207 [98.1%] vs 204 of 206 [99.0%]; difference, −0.9%; 95% CI, −3.6% to 1.8%; P = .72). More failed localizations occurred with the guidewire (21 of 208 [10.1%] vs 4 of 215 [1.9%]; difference, 8.2%; 95% CI, 3.3%-13.2%; P < .001). Median (IQR) time to specimen excision was shorter for the seed (15 [10-22] minutes vs 18 [12-30] minutes; P = .01), as was the total operative time (69 [56-86] minutes vs 75.5 [59-101] minutes; P = .03). The experience of surgeons, radiologists, and surgical coordinators was better with the seed.

Conclusions and Relevance  The combination of SPIO and a paramagnetic seed performed comparably with SPIO and guidewire for breast cancer conserving surgery and resulted in more successful localizations, shorter operative times, and better experience.

Trial Registration  ISRCTN.org Identifier: ISRCTN11914537

Background: Superparamagnetic iron nanoparticles perform comparably to radioisotope +/- blue dye for sentinel lymph node detection in breast cancer, even when injected up to 8 weeks before surgery. Using superparamagnetic iron nanoparticles for sentinel lymph node detection after primary systemic therapy, and the maximum time frame of superparamagnetic iron nanoparticle administration have not been investigated.

Methods: This cohort study included cN0/1-to-ycN0 patients undergoing sentinel lymph node detection or targeted axillary dissection. All patients received superparamagnetic iron nanoparticles either before primary systemic therapy or before surgery, and radioisotope on the day of surgery.

Results: For 113 patients analysed, superparamagnetic iron nanoparticles were injected a median of 3 (range 0-248) days before surgery, with a 97.4% detection rate compared with 91.2% for radioisotope (P = 0.057). Concordance for radioisotope was 97.1% and this was not affected by timing of superparamagnetic iron nanoparticle injection (Kendall's tau 0.027; P = 0.746). The median sentinel lymph node yield was 3 (interquartile range (i.q.r.) 2-3) for superparamagnetic iron nanoparticles and 2 (i.q.r. 2-3) for radioisotope (P < 0.001). In targeted axillary dissection, detection was 100% for superparamagnetic iron nanoparticles and 81.8% for radioisotope (P = 0.124). The index node was magnetic in 93.9% and radioactive in 66.7% (P = 0.007), an outcome that was not affected by any factors. For patients with metastases, superparamagnetic iron nanoparticle detection was 100% and radioisotope-based detection was 84.2% (P = 0.083), with superparamagnetic iron nanoparticles detecting more metastatic sentinel lymph nodes (median of 1 (i.q.r. 1-2) for superparamagnetic iron nanoparticles compared with a median of 1 (i.q.r. 0-1) for radioisotope; P = 0.005).

Conclusion: Injection before primary systemic therapy is feasible and does not affect concordance with radioisotope. Superparamagnetic iron nanoparticles perform comparably to radioisotope, but detect more sentinel lymph nodes and have a higher rate of detection of metastatic sentinel lymph nodes.

Simple Summary Immunotherapy blocking programmed cell death protein 1 (PD-1) and its ligands (PD-L1, PD-L2) is less effective in non-Hodgkin lymphoma (NHL) than classical Hodgkin lymphoma. However, NHL is a heterogeneous group and current research seeks to identify subgroups of NHL patients responsive to PD-1 blocking agents. Whether patients with pre-existing rheumatic diseases might constitute such a subgroup is unknown. We investigated the expression of PD-1 and its ligands in lymphoma patients with pre-existing rheumatic diseases. Our key findings include that in patients with pre-existing rheumatoid arthritis (RA) and subsequent diffuse large B-cell lymphoma, an association between RA disease severity and increased expression of PD-L1 in tumor cells was seen. This warrants further studies of the PD-1 pathway in lymphoma in other chronic inflammatory conditions. Current research seeks to identify subgroups of non-Hodgkin lymphoma (NHL) patients responsive to PD-1 blocking agents. Whether patients with pre-existing rheumatic diseases might constitute such a subgroup is unknown. We determined intratumoral expression of PD-1 and its ligands in lymphoma patients with pre-existing rheumatic diseases. We included 215 patients with rheumatoid arthritis (RA), systemic lupus erythematosus (SLE) or Sjogren's syndrome with subsequent lymphoma and 74 diffuse large B-cell lymphoma (DLBCL) controls without rheumatic disease. PD-1 and PD-ligand immunohistochemical markers were applied on tumor tissue microarrays. The number of PD-1+ tumor infiltrating leukocytes (TILs) and proportions of PD-L1+ and PD-L2+ tumor cells and TILs were calculated and correlated with clinical data. Expression of PD-L1 in tumor cells and TILs was highest in classical Hodgkin lymphoma and DLBCL. In DLBCLs, expression of PD-1 in TILs and PD-L1 in tumor cells was similar in RA, SLE and controls. In RA-DLBCL, high expression of PD-L1 in tumor cells was significantly more common in patients with the most severe RA disease and was associated with inferior overall survival in multivariable analysis.

Several autoimmune and inflammatory conditions, such as rheumatoid arthritis (RA) and primary Sjögrens’s syndrome (pSS), have repeatedly been linked to an increased risk of malignant lymphoma [1,2]. Certain inflammatory conditions are also associated with the development of specific lymphoma subtypes such as mucosa-associated lymphoid tissue (MALT) lymphoma in pSS and diffuse large B-cell lymphoma (DLBCL) in RA. The underlying mechanisms behind this association remain unclear. The highly increased risk of developing MALT lymphoma of the parotid gland in pSS indicates that local inflammatory processes can promote lymphoma development at the site of chronic inflammation [3]. In RA, an association between disease severity and risk of lymphoma has been shown.

Granulomatosis with polyangiitis (GPA), formerly Wegener’s granulomatosis, is a systemic small vessel vasculitis associated with the presence of anti-neutrophil cytoplasmic antibodies (ANCA) and characterized by granulomatous inflammation and necrotizing vasculitis of the airways and kidneys but possibly affecting any organ system. An increased risk of lymphoma in patients with GPA has been reported in several epidemiological studies [4,5]. However, very little is known about risk factors for lymphoma development in this group, possible relation to disease severity, treatment and lymphoma subtypes or the prognosis for the lymphomas. This is the first published study on GPA and lymphoma, giving detailed information on the GPA characteristics and possible risk factors for lymphoma and also lymphoma subtypes treatment and survival.