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Colloidal nanocrystals are miniscule pieces (< 100 nm or ∼ 1/1000th the width of a humanhair) of solid materials suspended in a liquid. Semiconductor nanocrystals have controllableoptical properties like color, which make them appealing for use in displays, LEDs, solarcells and many other devices that rely on the interaction of matter with light or electricity.The color and other optical properties are related to the size and composition of the crystals,which makes control over these properties crucial to any application.Creating these crystals is often more of an art than a science, with large differences inresults arising between chemicals sourced from different suppliers, laboratories, practitionersand any (even slight) changes in the process of making them. The work in this thesis ispart of a broader scientific movement aimed at a more scientific understanding and rationalcontrol of the synthesis process. To keep the nanocrystals small and stable requires the useof surfactants or ligands, which are chemicals that can interact both with a charged or polarenvironment such as the nanocrystal and a non-polar environment such as the surroundingmedium. The metal halide nanocrystals we focus on here are particularly sensitive to theseligands and the surrounding medium, as they are more similar in nature to salt, whereas mostother materials used to make nanocrystals are more similar to rocks. As a result, changingthe environment around the nanocrystals can transform the type, shape or composition ofthe crystal. The studies herein investigate the formation of metal halide nanocrystals, howtransformations between them occur and how both processes are interrelated.The rapid development in precision and scale of machine learning (ML) methods capableof predictions based on large sets of data has roused the interest of scientists across manydisciplines. Concurrently, improvements in the automation of chemical synthesis and charac-terization methods now allow for the generation of data on hundreds to thousands of reactionsnecessary to use such data science methods. Due to the complexity of chemical synthesis ingeneral, and nanocrystal synthesis in particular, using ML to predict the outcomes of reac-tions is an attractive proposition. This thesis introduces methods from machine learning anddata science to the synthesis of nanocrystals, and connects them to more physical models ofthis process.In this thesis, Chapter 1 outlines the important concepts related to nanocrystals, theirproperties, characterization methods and reactions. We then briefly introduce current trendsof using machine learning and data science to understand synthesis. In Chapter 2, weuse high-throughput synthesis experiments coupled to automated deconvolution of opticalabsorption spectra to gain an overview of the reaction landscape around one well-studiedmetal halide nanocrystal material, CsPbBr3. In Chapter 3, we investigate in more detailthe formation and transformation processes of one material we found of central importancein the overview, the atomically thin nanosheets of OLA2PbBr4 using in-situ absorptionspectroscopy and joint fits of spectral and kinetic models, as well as more traditional kineticanalysis where appropriate. Further analysis of precursor reactions is provided in Chapter 4.We then examine synthesis and properties of a family of cesium silver metal halide doubleperovskites in Chapter 5. Finally in Chapter 6, we discuss the implications of this work ondifferent scientific fields.

Background: The sarcoplasmic reticulum (SR) Ca 2+ -ATPase 2 (SERCA2) mediates Ca 2+ reuptake into SR and thereby promotes cardiomyocyte relaxation, whereas the ryanodine receptor (RYR) mediates Ca 2+ release from SR and triggers contraction. Ca 2+ /CaMKII (CaM [calmodulin]-dependent protein kinase II) regulates activities of SERCA2 through phosphorylation of PLN (phospholamban) and RYR through direct phosphorylation. However, the mechanisms for CaMKIIδ anchoring to SERCA2-PLN and RYR and its regulation by local Ca 2+ signals remain elusive. The objective of this study was to investigate CaMKIIδ anchoring and regulation at SERCA2-PLN and RYR. Methods: A role for AKAP18δ (A-kinase anchoring protein 18δ) in CaMKIIδ anchoring and regulation was analyzed by bioinformatics, peptide arrays, cell-permeant peptide technology, immunoprecipitations, pull downs, transfections, immunoblotting, proximity ligation, FRET-based CaMKII activity and ELISA-based assays, whole cell and SR vesicle fluorescence imaging, high-resolution microscopy, adenovirus transduction, adenoassociated virus injection, structural modeling, surface plasmon resonance, and alpha screen technology. Results: Our results show that AKAP18δ anchors and directly regulates CaMKIIδ activity at SERCA2-PLN and RYR, via 2 distinct AKAP18δ regions. An N-terminal region (AKAP18δ-N) inhibited CaMKIIδ through binding of a region homologous to the natural CaMKII inhibitor peptide and the Thr17-PLN region. AKAP18δ-N also bound CaM, introducing a second level of control. Conversely, AKAP18δ-C, which shares homology to neuronal CaMKIIα activator peptide (N2B-s), activated CaMKIIδ by lowering the apparent Ca 2+ threshold for kinase activation and inducing CaM trapping. While AKAP18δ-C facilitated faster Ca 2+ reuptake by SERCA2 and Ca 2+ release through RYR, AKAP18δ-N had opposite effects. We propose a model where the 2 unique AKAP18δ regions fine-tune Ca 2+ -frequency-dependent activation of CaMKIIδ at SERCA2-PLN and RYR. Conclusions: AKAP18δ anchors and functionally regulates CaMKII activity at PLN-SERCA2 and RYR, indicating a crucial role of AKAP18δ in regulation of the heartbeat. To our knowledge, this is the first protein shown to enhance CaMKII activity in heart and also the first AKAP (A-kinase anchoring protein) reported to anchor a CaMKII isoform, defining AKAP18δ also as a CaM-KAP.

Macrophages are critical for regulating inflammatory responses. Environmental signals polarize macrophages to either a proinflammatory (M1) state or an anti-inflammatory (M2) state. We observed that the microRNA (miRNA) cluster mirn23a, coding for miRs-23a, -27a, and -24-2, regulates mouse macrophage polarization. Gene expression analysis of mirn23a-deficient myeloid progenitors revealed a decrease in TLR and IFN signaling. Mirn23a−/− bone marrow–derived macrophages (BMDMs) have an attenuated response to LPS, demonstrating an anti-inflammatory phenotype in mature cells. In vitro, mirn23a−/− BMDMs have decreased M1 responses and an enhanced M2 responses. Overexpression of mirn23a has the opposite effect, enhancing M1 and inhibiting M2 gene expression. Interestingly, expression of mirn23a miRNAs goes down with inflammatory stimulation and up with anti-inflammatory stimulation, suggesting that its regulation prevents locking macrophages into polarized states. M2 polarization of tumor-associated macrophages (TAMs) correlates with poor outcome for many tumors, so to determine if there was a functional consequence of mirn23a loss modulating immune cell polarization, we assayed syngeneic tumor growth in wild-type and mirn23a−/− mice. Consistent with the increased anti-inflammatory/immunosuppressive phenotype in vitro, mirn23a−/− mice inoculated with syngeneic tumor cells had worse outcomes compared with wild-type mice. Coinjecting tumor cells with mirn23a−/− BMDMs into wild-type mice phenocopied tumor growth in mirn23a−/− mice, supporting a critical role for mirn23a miRNAs in macrophage-mediated tumor immunity. Our data demonstrate that mirn23a regulates M1/M2 polarization and suggests that manipulation of mirn23a miRNA can be used to direct macrophage polarization to drive a desired immune response.

Fibrosis accompanies most heart diseases and is associated with adverse patient outcomes. Transforming growth factor (TGF)β drives extracellular matrix remodelling and fibrosis in the failing heart. Some members of the ADAMTSL (a disintegrin-like and metalloproteinase domain with thrombospondin type 1 motifs-like) family of secreted glycoproteins bind to matrix microfibrils, and although their function in the heart remains largely unknown, they are suggested to regulate TGFβ activity. The aims of this study were to determine ADAMTSL2 levels in failing hearts, and to elucidate the role of ADAMTSL2 in fibrosis using cultured human cardiac fibroblasts (CFBs). Cardiac ADAMTSL2 mRNA was robustly increased in human and experimental heart failure, and mainly expressed by fibroblasts. Over-expression and treatment with extracellular ADAMTSL2 in human CFBs led to reduced TGFβ production and signalling. Increased ADAMTSL2 attenuated myofibroblast differentiation, with reduced expression of the signature molecules α-smooth muscle actin and osteopontin. Finally, ADAMTSL2 mitigated the pro-fibrotic CFB phenotypes, proliferation, migration and contractility. In conclusion, the extracellular matrix-localized glycoprotein ADAMTSL2 was upregulated in fibrotic and failing hearts of patients and mice. We identified ADAMTSL2 as a negative regulator of TGFβ in human cardiac fibroblasts, inhibiting myofibroblast differentiation and pro-fibrotic properties.

There are large between- and within-country variations in COVID-19 death rates. Some very low death rate settings such as Eastern Asia, Central Europe, the Balkans and Africa have a common feature of eating large quantities of fermented foods whose intake is associated with the activation of the Nrf2 (Nuclear factor (erythroid-derived 2)-like 2) anti-oxidant transcription factor. There are many Nrf2-interacting nutrients (berberine, curcumin, epigallocatechin gallate, genistein, quercetin, resveratrol, sulforaphane) that all act similarly to reduce insulin resistance, endothelial damage, lung injury and cytokine storm. They also act on the same mechanisms (mTOR: Mammalian target of rapamycin, PPARγ:Peroxisome proliferator-activated receptor, NFκB: Nuclear factor kappa B, ERK: Extracellular signal-regulated kinases and eIF2α:Elongation initiation factor 2α). They may as a result be important in mitigating the severity of COVID-19, acting through the endoplasmic reticulum stress or ACE-Angiotensin-II-AT1R axis (AT1R) pathway. Many Nrf2-interacting nutrients are also interacting with TRPA1 and/or TRPV1. Interestingly, geographical areas with very low COVID-19 mortality are those with the lowest prevalence of obesity (Sub-Saharan Africa and Asia). It is tempting to propose that Nrf2-interacting foods and nutrients can re-balance insulin resistance and have a significant effect on COVID-19 severity. It is therefore possible that the intake of these foods may restore an optimal natural balance for the Nrf2 pathway and may be of interest in the mitigation of COVID-19 severity.

OBJECTIVE: In the phase 3 VELIA/GOG-3005 trial, veliparib added to carboplatin-paclitaxel and continued as maintenance improved progression-free survival (PFS) compared to carboplatin-paclitaxel alone in patients with newly diagnosed ovarian carcinoma. Primary analysis of PFS was by investigator (INV) assessment, with a supplemental analysis of PFS by blinded independent central review (BICR).METHODS: Patients received veliparib or placebo with carboplatin-paclitaxel (6 cycles) and as maintenance (30 additional cycles). The primary analysis compared PFS in the veliparib-throughout arm to the carboplatin-paclitaxel only arm in the BRCA mutation (BRCAm), homologous recombination deficiency (HRD), and intention-to-treat (ITT) populations. Exploratory analyses of PFS in BRCA wildtype (BRCAwt), homologous recombination proficient (HRP), and HRD + BRCAwt populations were also performed. PFS per BICR and overall concordance rates between INV and BICR assessments were analyzed.RESULTS: Hazard ratios for PFS by INV and BICR were consistent in each of the primary analysis and exploratory populations. In the ITT population, median PFS per INV was 23.5 months in the veliparib-throughout arm versus 17.3 months in the control arm (hazard ratio [HR] 0.683, 95% confidence interval [CI] 0.562-0.831; P CONCLUSIONS: Hazard ratios for PFS per BICR and per INV were consistent, with no suggestion of investigator bias. These findings support the reliability of PFS by INV in ovarian cancer trials.

A hallmark of the pancreatic hormone amylin is its high propensity toward the formation of amyloid fibrils, which makes it a challenging drug design effort. The amylin analogue pramlintide is commercially available for diabetes treatment as an adjunct to insulin therapy but requires three daily injections due to its short half-life. We report here the development of the stable, lipidated long-acting amylin analogue cagrilintide (23) and some of the structure-activity efforts that led to the selection of this analogue for clinical development with obesity as an indication. Cagrilintide is currently in clinical trial and has induced significant weight loss when dosed alone or in combination with the GLP-1 analogue semaglutide.

Roundup Transorb® (RDT) is a glyphosate-based herbicide commonly used in agricultural practices worldwide. This herbicide exerts negative effects on the aquatic ecosystem and affects bioenergetic and detoxification pathways, oxidative stress, and cell damage in marine organisms. These effects might also occur at the transcriptional level; however, the expression of genes associated with oxidative stress has not been studied well. Odontesthes humensis is a native Brazilian aquatic species naturally distributed in the habitats affected by pesticides, including Roundup Transorb® (RDT). This study evaluated the toxic effects of short-term exposure to RDT on O. humensis. Moreover, the genes related to oxidative stress were sequenced and characterized, and their expressions in the gills, hepatopancreas, kidneys, and brain of the fish were quantified by quantitative reverse transcription-polymerase chain reaction. The animals were exposed to two environmentally relevant concentrations of RDT (2.07 and 3.68 mg L−1) for 24 h. Lipid peroxidation, reactive oxygen species (ROS), DNA damage, and apoptosis in erythrocytes were quantified by flow cytometry. The expression of the target genes was modulated in most tissues in the presence of the highest tested concentration of RDT. In erythrocytes, the levels of lipid peroxidation, ROS, and DNA damage were increased in the presence of both the concentrations of RDT, whereas cell apoptosis was increased in the group exposed to 3.68 mg L−1 RDT. In conclusion, acute exposure to RDT caused oxidative stress in the fish, induced negative effects on cells, and modulated the expression of genes related to the enzymatic antioxidant system in O. humensis. Fil: Martins, Amanda Weege S.. Universidade Federal de Pelotas; Brasil Fil: Silveira, Tony L. R.. Universidade Federal de Pelotas; Brasil. Universidade Federal do Rio Grande; Brasil Fil: Remião, Mariana H.. Universidade Federal de Pelotas; Brasil Fil: Domingues, William Borges. Universidade Federal de Pelotas; Brasil Fil: Dellagostin, Eduardo N.. Universidade Federal de Pelotas; Brasil Fil: Varela Junior, Antônio Sergio. Universidade Federal do Rio Grande; Brasil. Universidade Federal de Pelotas; Brasil Fil: Corcini, Carine D.. Universidade Federal de Pelotas; Brasil Fil: Costa, Patrícia G.. Universidade Federal do Rio Grande; Brasil Fil: Bianchini, Adalto. Universidade Federal do Rio Grande; Brasil Fil: Somoza, Gustavo Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina Fil: Robaldo, Ricardo B.. Universidade Federal de Pelotas; Brasil Fil: Campos, Vinicius Farias. Universidade Federal de Pelotas; Brasil

Real-time super-resolution microscopy analysis reveals the growth kinetics, morphology, and abundance of human insulin amyloid spherulites with different growth pathways.Protein misfolding in the form of fibrils or spherulites is involved in a spectrum of pathological abnormalities. Our current understanding of protein aggregation mechanisms has primarily relied on the use of spectrometric methods to determine the average growth rates and diffraction-limited microscopes with low temporal resolution to observe the large-scale morphologies of intermediates. We developed a REal-time kinetics via binding and Photobleaching LOcalization Microscopy (REPLOM) super-resolution method to directly observe and quantify the existence and abundance of diverse aggregate morphologies of human insulin, below the diffraction limit and extract their heterogeneous growth kinetics. Our results revealed that even the growth of microscopically identical aggregates, e.g., amyloid spherulites, may follow distinct pathways. Specifically, spherulites do not exclusively grow isotropically but, surprisingly, may also grow anisotropically, following similar pathways as reported for minerals and polymers. Combining our technique with machine learning approaches, we associated growth rates to specific morphological transitions and provided energy barriers and the energy landscape at the level of single aggregate morphology. Our unifying framework for the detection and analysis of spherulite growth can be extended to other self-assembled systems characterized by a high degree of heterogeneity, disentangling the broad spectrum of diverse morphologies at the single-molecule level.

Electrophilic 11 C-labelled aroyl dimethylaminopyridinium salts, obtained by carbonylative cross-coupling of aryl halides with [11 C]carbon monoxide, were prepared for the first time and shown to be valuable intermediates in the synthesis of primary [11 C]benzamides. The methodology furnished a set of benzamide model compounds, including the two poly (ADP-ribose) polymerase (PARP) inhibitors niraparib and veliparib, in moderate to excellent radiochemical yields. In addition to providing a convenient and practical route to primary [11 C]benzamides, the current method paves the way for future application of [11 C]aroyl dimethylaminopyridinium halide salts in positron emission tomography (PET) tracer synthesis.

We report herein an efficient and fully automated protocol for the radiosynthesis of [18 F]FAPI-74, a new positron emission tomography (PET) radiopharmaceutical for in vivo detection of the fibroblast activation protein. [18 F]FAPI-74 was synthesized via a rapid [18 F]aluminum fluoride coordination reaction, which was first developed on the flexible GE TRACERLab FX2N (FXN) platform and later translated to the cassette-based module Trasis AllInOne (AIO). The results obtained with both modules were comparable in terms of yield and reproducibility. Automation of [18 F]FAPI-74 radiosynthesis on the FXN was carried out in 35 min with a radiochemical yield (RCY) of 18.5 ± 2.5% (n = 5, relative to starting [18 F]fluoride). Method transfer to the AIO platform following minor optimizations allowed for the production of [18 F]FAPI-74 in an isolated RCY of 20 ± 2.5% [n = 3] with an overall synthesis time of 40 min. The radiochemical purity was greater than 95% for [18 F]FAPI-74, obtained from both modules. Overall, the protocol reliably provides a sterile and pyrogen-free good manufacturing practice (GMP) compliant product of [18 F]FAPI-74 suitable for clinical PET imaging.

Objective: Sepsis causes millions of deaths on a global scale annually. Activation of peptidylarginine deiminase (PAD) enzymes in sepsis causes citrullination of histones, which results in neutroph...

In this study, the toxic effects of melittin on Madin-Darby Bovine Kidney cells (MDBK) were analyzed with respect to mitochondrial functionality by reduction of MTT and flow cytometry, apoptosis potential, necrosis, oxygen reactive species (ROS) production, lipid peroxidation, and DNA fragmentation using flow cytometry and cell membrane destabilization by confocal microscopy. The toxicity presented dose-dependent characteristics and mitochondrial activity was inhibited by up to 78.24 ±3.59% (P

The large natural gas storage facility at Stenlille, Denmark, has been monitored to investigate the effect of pumping large amounts of gas into the subsurface. Here, we present a new dataset of microseismicity at Stenlille since 2018. We compare these data with methane in groundwater, which has been monitored since gas storage was established in 1989. Further, we conducted a controlled 172 day microcosm experiment of methane oxidation on an isolated microbial community under both aerobic and anaerobic conditions. For this experiment, water was filtered from a well at Stenlille with elevated levels of thermogenic methane and ethane. No microseismic activity was detected in the gas storage area above an estimated detection level of ML 0.0 for the established network. The long-term monitoring for methane in groundwater has still only detected one leak, in 1995, related to a technical problem during injection. The microcosm experiment revealed that oxidation of methane occurred only under aerobic conditions during the experiment, as compared to anaerobic conditions, even though the filtered water was anoxic

Data suggest that renal denervation (RDN) in treatment-resistant hypertension (TRHT) is safe in terms of renal function. However, most studies report kidney function as creatinine-based estimated glomerular filtration rate (eGFR), which may be biased by non-renal factors. Damage markers other than albuminuria have never been evaluated after RDN. In this non-randomized RDN trial, we studied changes in kidney function, assessed as measured GFR (mGFR) and various GFR estimates, six months and two years after RDN. We also examined changes in albuminuria and a biomarker of tubular dysfunction. Adult non-diabetic patients with TRHT and eGFR ≥45 ml/min/1.73 m2 were recruited from hypertension clinics. Before bilateral RDN, mGFR was measured by iohexol clearance. We estimated eGFR from serum creatinine and cystatin C (eGFRcrea, eGFRcys, and eGFRcreacys), and albumin-creatinine ratio (ACR) and N-acetyl-β-D-glucosaminidase (NAG)-creatinine ratio (NAG-CR) were measured in spot urines. All measurements were repeated after six and twenty-four months. Twenty patients, mean age 54 (±9) years and baseline mGFR 83 (±20) ml/min/1.73 m2 underwent RDN. After six months, mGFR fell, eGFRcrea remained unchanged, whereas eGFRcys and eGFRcreacys increased. At 2 years’ follow-up, eGFRcreacys was significantly lower than at baseline. mGFR was 78 (±28) ml/min/1.73 m2. Change in ambulatory systolic BP predicted change in eGFRcrea. Urinary NAG-CR, but not ACR, increased during follow-up. Different GFR assessments gave diverging results after RDN. Therefore, care should be taken to method when evaluating kidney function after RDN. Increases in a tubular dysfunction biomarker suggest that kidney damage may occur. Long-term renal follow-up is needed after RDN.

The synthesis quality of artificial inorganic nanocrystals is most often assessed by transmission electron microscopy (TEM) for which high-throughput advances have dramatically increased both the quantity and information richness of metal nanoparticle (mNP) characterization. Existing automated data analysis algorithms of TEM mNP images generally adopt a supervised approach, requiring a significant effort in human preparation of labeled data that reduces objectivity, efficiency, and generalizability. We have developed an unsupervised algorithm AutoDetect-mNP for automated analysis of TEM images that objectively extracts morphological information on convex mNPs from TEM images based on their shape attributes, requiring little to no human input in the process. The performance of AutoDetect-mNP is tested on two data sets of bright field TEM images of Au nanoparticles with different shapes and further extended to palladium nanocubes and cadmium selenide quantum dots, demonstrating that the algorithm is quantitatively reliable and can thus serve as a generalizable measure of the morphology distributions of any mNP synthesis. The AutoDetect-mNP algorithm will aid in future developments of high-throughput characterization of mNPs and the future advent of time-resolved TEM studies that can investigate reaction mechanisms of mNP synthesis and reactivity.

Lifetime and reliability in realistic operating conditions are important parameters for the application of thin-film piezoelectric microelectromechanical systems (piezoMEMS) based on lead zirconate titanate (PZT). Humidity can induce time-dependent dielectric breakdown at a higher rate compared to dry conditions, and significantly alter the dynamic behavior of piezoMEMS-devices. Here we assess the lifetime and reliability of PZT-based micromirrors with and without humidity barriers operated at 23°C in an ambient of 0 and 95 % relative humidity. The correlation of the dynamic response, as well as the ferroelectric, dielectric, and leakage properties, with degradation time was investigated. In humid conditions, the median time-to-failure was increased from ${2.7\times }{10}^{4}{[1.9\times }{10}^{4}{-4.0\times }{10}^{4}{]}$ s to ${1.1\times }{10}^{6}{[0.9\times }{10}^{6}{-1.5\times }{10}^{6}{]}$ s at 20 $\text{V}_{{\mathrm {AC}}}$ continuous unipolar actuation, by using a 40 nm thick Al2O3 humidity barrier. However, the initial maximum angular deflection, polarization, and dielectric permittivity decreased by about 6, 11, and 12 %, respectively, for Al2O3 capped devices. For both bare and encapsulated devices, the onset of electrothermal breakdown-events was the dominant cause of degradation. Severe distortions in the device’s dynamic behavior, together with failure from loss of angular deflection, preceded time-dependent dielectric breakdown in 95% relative humidity. Moreover, due to the film-substrate stress transfer sensitivity of thin-film devices, water-induced degradation affects the reliability of thin-film piezoMEMS differently than bulk piezoMEMS. [2020-0228]

Background: Mild-to-moderate iodine deficiency is still present in many countries, particularly in pregnant women. Observational studies suggest that mild-to-moderate iodine deficiency during pregnancy may be associated with impaired thyroid function and child neurodevelopment. Randomized-controlled food trials to increase iodine status are scarce. We assessed the impact of an increased intake of cod during pregnancy on maternal iodine status and infant neurodevelopment. Methods: In this randomized-controlled trial, pregnant women in Bergen, Norway, recruited through Haukeland University Hospital, were randomly assigned (1:1) to an intervention of 200 g of cod twice a week for 16 weeks (gestational week 20–36) or to continue with their standard diet (control group). Randomization was done by lottery. Primary outcome was urinary iodine concentration (UIC) (spot samples from six consecutive days) measured postintervention. Secondary outcome was infant neurodevelopment assessed by the cognitive, language, and motor scales of the Bayley Scales of Infant and Toddler Developmental third edition (Bayley-III) at 11 months of age. In addition, maternal thyroid function was measured (thyrotropin [TSH], free triiodothyronine [fT3], free thyroxine [fT4]) at baseline and postintervention. Results: Between January 2016 until February 2017, 137 women were recruited. Postintervention UIC was higher in the intervention group (n = 61) [median (interquartile range, IQR) 98 (64–145) μg/L], compared with control (n = 61) [median (IQR) 73 (52–120) μg/L] (p = 0.028), also after adjusting for baseline UIC (p = 0.048). Infants of mothers in the intervention group had a lower cognitive composite score on the Bayley-III compared with the control group (p = 0.045). There were no group differences in the Bayley III language- or motor composite scores. Maternal thyroid hormones (TSH, fT3, fT4) did not differ between the groups postintervention. Conclusions: Increased cod intake during pregnancy improved the iodine status in women with mild-to-moderate iodine deficiency, however, did not affect thyroid function. The negative effect on cognition should be followed up to assess whether this is a stable effect over time. More studies are warranted to enable good health advice on iodine nutrition in pregnancy. ClinicalTrials.gov NCT02610959. Registered November 20, 2015.

All immunological methods are vulnerable to different kinds of interference. The purpose of this work was to study the cause and frequency of method-dependent interference in the Roche thyroid immunoassays. Serum samples with discordant thyroid function tests (TFT) were selected from samples sent to the Hormone Laboratory, Oslo University Hospital from June 2013 to September 2018. We identified 93 patients with discordant pathological TFT when analysed with the Roche methods and normal results when analysed with alternative methods. 42 of these samples were sent to Roche Diagnostics (Germany) for investigation of the interfering substance. Roche found interference to be caused by the presence of endogenous anti-streptavidin antibodies (ASA) (34 of 42 patients), ruthenium or the idiotype of the ruthenium labelled antibody (3 of 42 patients) and mouse antigens (1 of 42 patients). Method-dependent interference was estimated to affect 0.37% of the patients investigated in our laboratory. Interference due to the presence of endogenous ASA were further explored in other (non-thyroid) immunoassays by comparing analyte levels before and after pre-adsorption of the patients’ sera with streptavidin-coated paramagnetic beads. An underestimation of hormone levels was observed in sandwich immunoassays, while an overestimation was found in competitive assays. Method-dependent interference in Roche thyroid immunoassays is caused mainly by ASA and is not a very rare phenomenon. Misleading results may lead to misdiagnosis and inappropriate medical treatment. The supplier of the assay should be alerted when the available alternative methodology reveals method-dependent errors.

Exposure of isolated human islets to proinflammatory cytokines leads to up-regulation of inducible nitric oxide synthase (iNOS), raised NO, and beta cell toxicity. These findings have led to increasing interest in the clinical utility of iNOS blockade to mitigate beta cell destruction in human type 1 diabetes (T1D). However, recent studies show that iNOS-derived NO may also confer beta cell protection. To investigate this dichotomy, we compared islet cell distributions and intensity of iNOS immunostaining in pancreatic sections, co-stained for insulin and glucagon, from new-onset T1D donors (group 1), with non-diabetic autoantibody-negative (group 2), non-diabetic autoantibody-positive (group 3) and long-term diabetic donors (group 4). The cellular origins of iNOS, its frequency and graded intensities in islets and number in peri-islet, intra-islet and exocrine regions were determined. All donors showed iNOS positivity, irrespective of disease and presence of beta cells, had variable labelling intensities, without significant differences in the frequency of iNOS-positive islets among study groups. iNOS was co-localised in selective beta, alpha and other endocrine cells, and in beta cell-negative islets of diabetic donors. The number of peri- and intra-islet iNOS cells was low, being significantly higher in the peri-islet area. Exocrine iNOS cells also remained low, but were much lower in group 1. We demonstrate that iNOS expression in islet cells is variable, heterogeneous and independent of co-existing beta cells. Its distribution and staining intensities in islets and extra-islet areas do not correlate with T1D or its duration. Interventions to inactivate the enzyme to alleviate disease are currently not justified.

By screening of a collection of 50 000 small-molecule compounds, we recently identified 4-arylazo-3,5-diamino-1H-pyrazoles as a novel group of anti-biofilm agents. Here, we report a SAR study based on 60 analogues by examining ways in which the pharmacophore can be further optimized, for example, via substitutions in the aryl ring. The SAR study revealed the very potent anti-biofilm compound 4-(2-(2-fluorophenyl)hydrazineylidene)-5-imino-4,5-dihydro-1H-pyrazol-3-amine (2).

Atmospheric particles negatively affect human health, limit visibility and impact climate. Their deleterious effects are known, but the mechanism of how particles form and grow in the atmosphere is not fully understood. Sulfuric acid (H2SO4) is a large contributor to particle formation, but nucleation of H2SO4-H2O cannot explain atmospheric observations. Ammonia (NH3) can form particles with sulfuric acid, yet the ternary nucleation of H2SO4-NH3-H2O still does not match measurements, indicating the participation of other precursors. Amines have been shown to enhance particle formation and, despite their lower concentrations in air, can displace NH3 in clusters. As the use of sulfur-based fossil fuels is phased out, and consequently H2SO4 concentrations are reduced, methanesulfonic acid (MSA) is expected to become a more important source of particles. MSA does not form particles efficiently with water alone, but does so with amines and water. The reaction of MSA and ammonia/amines is dependent on relative humidity, basicity and amine structure. In this dissertation, the effect of four organic compounds on the reaction of methanesulfonic acid, amines and water is investigated. Experiments are conducted in a small volume aerosol flow reactor at ambient temperature and atmospheric pressure (294 K, 1 atm). Early experiments show that the aerosol flow reactor is sensitive to order of addition of reactants. Laboratory results are interpreted along with results of theoretical calculations of initial clusters completed by the Gerber group. Results show that particle formation is influenced by proton transfer, hydrogen bonding capacity and basicity. Molecular structure can also impact particle formation and growth by influencing how initial clusters grow to detectable particles. Particle formation from organics and amines with and without water is inefficient in this system. Results show that, in the atmosphere, water likely overwhelms the effect of organics. Theoretical calculations give insight on how experimentally unobservable initial clusters of these systems correlates with detectable particles. The results of this work could aid atmospheric models in more accurately predicting the impact of particles on a regional and global level.

The Norwegian Scientific Committee for Food Safety (Vitenskapskomiteen for mattrygghet, VKM) has on the request from the Norwegian Food Safety Authority (Mattilsynet) assessed four studies on developmental neurotoxicity following low dose exposure to bisphenol A (BPA) (Adriani et al., 2003; Carr et al., 2003; Negishi et al., 2004; Ryan and Vandenbergh, 2006). The background for the request is uncertainties related to developmental neurotoxcity of BPA raised by the Nordic environmental agencies in Norway, Sweden and Denmark. VKM was asked to consider if these studies provide sufficient evidence to set a lower no observed adverse effect level (NOAEL) in the hazard characterisation of BPA. Further, a Norwegian exposure scenario based on available exposure data should be performed. The task has been assessed by the Scientific Panel on Food Additives, Flavourings, Processing Aids, Materials in Contact with Food and Cosmetics (Panel 4). Bisphenol A (CAS number 80-05-7) is primarily used as a monomer in the production of polycarbonate, which is used to make food containers, such as beverage bottles, baby bottles, tableware and storage containers. It is also used as a precursor of certain epoxy resins used for protective coatings for food and beverage cans. BPA is permitted for use in food contact materials in the European Union (EU) with a specific migration limit (SML) of 0.6 mg/kg food. The migration limit in the EU regulations has yet to be modified according to an opinion from the European Food Safety Authority (EFSA) from 2006 where a new established tolerable daily intake (TDI) of 0.05 mg BPA/kg body weight (bw) was derived from a NOAEL of 5 mg/kg bw/day. A European Union Risk Assessment Report (RAR) of BPA produced in accordance with Council Regulation (EEC) 793/93 has recently been updated (April 2008) reviewing a previously requested 2-generation study in mice (Tyl et al., 2007) and new data on human exposure and effects of BPA. A NOAEL of 50 mg/kg bw/day was suggested in this report. The Nordic environmental agencies (Norway, Sweden and Denmark) have participated in the discussions on this updated EU RAR of BPA and they strongly disagreed that this NOAEL also covers developmental neurotoxicity. According to the Nordic environmental agencies, the four above mentioned studies indicate a possible risk for developmental neurotoxicity of BPA at very low exposure levels (0.1-0.25 mg/kg bw/day). The position of the Nordic environmental agencies has been included as a footnote in the revised EU RAR. Recently, in April 2008, the U.S. National Toxicology Program (NTP), Health Canada and Environment Canada have published draft reports on effects of BPA, including developmental effects (neural and behavioural effects) and expressed some concern for neural and behavioural effects in fetuses, infants and children at current human exposures. The European Commission has therefore asked EFSA to further assess possible age dependent toxicokinetics of BPA in animals and humans and their implications for hazard and risk assessment of BPA taken into account the most recent information and data available. The present opinion from VKM Panel 4 is based on an evaluation of the design, conduct (or accomplishment) and the results in the four above mentioned studies. The study design has been evaluated in light of international recommendations given in relevant guidelines dealing with developmental neurotoxicity testing in animals. The recent international developments on BPA in the U.S. and Canada are not addressed in this opinion. The report by Tyl and co-workers was central in the EFSA opinion from 2006 and the updated EU RAR from 2008. The Tyl study is a GLP compliant 2-generation reproductive toxicity evaluation in mice performed according to a modified OECD 416 guideline. However, the study did not include functional tests for developmental neurotoxicity. VKM has reviewed the four above mentioned studies on neurodevelopmental toxicity of BPA as requested by the Norwegian Food Safety Authority. Although the design and reporting of these studies suffer from major and serious shortcomings, the overall findings may raise some concern. It is the opinion of the VKM Panel 4 that the four studies do not provide sufficient evidence for setting a robust lower NOAEL for BPA than the current EFSA NOAEL of 5 mg/kg bw/day. The Panel is aware that the EU Commission recently has requested EFSA to re-evaluate the information available on BPA. In order to eliminate any uncertainty regarding potential developmental effects of BPA at low doses, it is recommended that a GLP compliant study is carried out according to OECD guideline 426. Such a study should utilize a broad concentration range from the very low doses up to those with known maternal effects. A Norwegian exposure scenario based on available data on exposure to BPA from food and beverages and via the environment was performed. In general, exposure levels of BPA in Norway are low. The estimated exposure of infants and children is in the range of 3.5 – 13.2 μg/kg bw/day, whereas the estimated aggregated exposure of adults is 1.5 μg/kg bw/day. As a result of the current use of BPA in food contact materials and other consumer products, infants and children are exposed to higher levels of BPA per kg body weight than the rest of the population.

Among the immune system cells, macrophages have an important role. Apamin, a bee venom constituent, is important in the defense of these insects. Thus, we aimed to evaluate the metabolism of J774 1.6 macrophage cell line when exposed to isolated and purified apamin, using cytotoxicity tests by MTT reduction and analysis by flow cytometry (apoptosis / necrosis, production of reactive oxygen species (ROS), membranous lipoperoxidation (LPO), electrical potential of the mitochondrial membrane (mMP) and DNA fragmentation). None of the tested concentrations (10 to 100μg/mL) were cytotoxic according to MTT reductions. Apoptosis rates decreased at concentrations of 2.5, 5.0, and 10.0μg/mL (P0.05). Apamin did not alter ROS, LPO, or DNA fragmentation. Therefore, all analyzed concentrations (1.25 to 10μg/mL) decreased mMP. Such decrease in apoptosis might be due to a suppression of mitochondrial pro-apoptotic messengers, as this peptide causes no oxidative stress, lipid peroxidation, and DNA damage. Highly sensitive techniques are majorly important for proper interpretation of cellular toxicity mechanisms, combined with routine laboratory methods. RESUMO Das células do sistema imunológico, macrófagos desempenham um papel fundamental. Apamina, constituinte do veneno de abelhas, é importante na defesa destas. Objetivou-se avaliar o metabolismo da linhagem de macrófagos J774 1.6 expostos à apamina isolada e purificada, avaliando-se citotoxicidade por redução de MTT e análise por citometria de fluxo (apoptose / necrose, produção de espécies reativas de oxigênio (EROs), lipoperoxidação membranosa (LPO), potencial elétrico da membrana mitocondrial (MMP) e fragmentação do DNA). Nenhuma concentração testada (10 a 100μg / mL) foi citotóxica. As taxas de apoptose diminuíram nas concentrações 2,5, 5,0 e 10,0μg / mL (P0,05). A apamina não alterou as ERO, a LPO nem a fragmentação do DNA. Portanto, todas as concentrações analisadas (1,25 a 10μg / mL) diminuíram a mMP. Tal diminuição na apoptose pode ser por uma supressão de mensageiros pró-apoptóticos mitocondriais, já que este peptídeo não causa estresse oxidativo, peroxidação lipídica nem dano ao DNA. Técnicas altamente sensíveis são importantes para adequada interpretação dos mecanismos de citotoxicidade.

Background More than 170 post-transcriptional RNA modifications regulate the localization, processing and function of cellular RNAs, and aberrant RNA modifications have been linked to a range of human diseases. The RNA modification landscape in atherosclerosis, the main underlying cause of cardiovascular diseases, is still largely unknown. Methods We used mass spectrometry to analyse a selection of RNA-modifying enzymes and the N6-methyladenosine (m6A) in carotid atherosclerotic lesion samples representing early and advanced stages of atherosclerosis as compared to non-atherosclerotic arteries from healthy controls. Findings (i) the detection of different levels of several enzymes involved in methylations occurring in rRNA and mRNA; (ii) these findings included changes in the levels of methyltransferases (‘writers’), binding proteins (‘readers’) and demethylases (‘erasers’) during atherosclerosis as compared to non-atherosclerotic control arteries, with generally the most prominent differences in samples from early atherosclerotic lesions; and (iii) these changes were accompanied by a marked downregulation of m6A in rRNA, the most abundant and well-studied modification in mRNA with a wide range of effects on cell biology. Interpretation We show for the first time that RNA-modifying enzymes and the well-studied RNA modification m6A are differentially regulated in atherosclerotic lesions, which potentially could help creating new prognostic and treatment strategies.

Type 2 diabetes (T2D), alike Parkinson’s disease (PD), belongs to the group of protein misfolding diseases (PMDs), which share aggregation of misfolded proteins as a hallmark. Although the major aggregating peptide in β-cells of T2D patients is Islet Amyloid Polypeptide (IAPP), alpha-synuclein (αSyn), the aggregating peptide in substantia nigra neurons of PD patients, is expressed also in β-cells. Here we show that αSyn, encoded by Snca, is a component of amyloid extracted from pancreas of transgenic mice overexpressing human IAPP (denoted hIAPPtg mice) and from islets of T2D individuals. Notably, αSyn dose-dependently promoted IAPP fibril formation in vitro and tail-vein injection of αSyn in hIAPPtg mice enhanced β-cell amyloid formation in vivo whereas β-cell amyloid formation was reduced in hIAPPtg mice on a Snca −/− background. Taken together, our findings provide evidence that αSyn and IAPP co-aggregate both in vitro and in vivo, suggesting a role for αSyn in β-cell amyloid formation.

We describe the design of a novel PDE4 scaffold and the exploration of the dual-soft concept to reduce systemic side effects via rapid elimination: introducing ester functionalities that can be inactivated in blood as well as by the liver (dual-soft) while being stable in human skin. Compound 40 was selected as a clinical candidate as it was potent and rapidly degraded by blood and liver to inactive metabolites and because in preclinical studies it showed high exposure at the target organ: the skin. Preclinical and clinical data are presented confirming the value of the dual-soft concept in reducing systemic exposure.

Tiny samples of ancient atmosphere in air bubbles within ice cores contain argon (Ar), which can be used to reconstruct past temperature changes. At a sufficient depth, the air bubbles are compressed by the overburden pressure under low temperature and transform into air-hydrate crystals. While the oxygen (O2) and nitrogen (N2) molecules have indeed been identified in the air-hydrate crystals with Raman spectroscopy, direct observational knowledge of the distribution of Ar at depth within ice sheet and its enclathration has been lacking. In this study, we applied scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDS) to five air-hydrate crystals in the Greenland NEEM ice core, finding them to contain Ar and N. Given that Ar cannot be detected by Raman spectroscopy, the method commonly used for O2 and N2, the SEM-EDS measurement method may become increasingly useful for measuring inert gases in deep ice cores.

The noradrenergic locus coeruleus (LC) is a small brainstem nucleus that promotes arousal and attention. Recent studies have examined the microstructural properties of the LC using diffusion-weighted magnetic resonance imaging and found unexpected age-related differences in fractional anisotropy - a measure of white matter integrity. Here, we used three datasets (Berlin Aging Study-II, N = 301, the Leipzig Study for Mind-Body-Emotion Interactions, N = 220, and Stockholm Sleepy Brain, N = 49), to replicate published findings and expand them by investigating diffusivity in the LC’s ascending noradrenergic bundle. In younger adults, LC fractional anisotropy was significantly lower, compared to older adults. However, in the LC’s ascending noradrenergic bundle, we observed significantly higher fractional anisotropy in younger adults, relative to older adults. These findings indicate that diffusivity in the LC versus the ascending noradrenergic bundle are both susceptible to microstructural changes in aging that have opposing effects on fractional anisotropy.HighlightsFractional anisotropy in the locus coeruleus was lower in younger adultsFractional anisotropy in the noradrenergic bundle was higher in younger adultsSleep deprivation may affect diffusivity in younger adults more than older adults

Incineration is in many countries a common treatment method for municipal solid waste, and utilization of the ash residues has attracted significant interest. The bottom ash is best suited as a secondary construction material, whereas the fly ash is being investigated as a secondary raw material for recovery of, for example, Zn, Cu, and salts. For both types of application, knowledge about the chemical speciation of Zn and Cu in the ashes is valuable. The present work focuses on identifying and quantifying the chemical species of Zn and Cu in 12 samples of fly ash and bottom ash from three waste-to-energy plants using X-ray absorption near edge structure (XANES). The XANES spectra of the ash samples showed similar distinctive features, and both in the bottom and fly ash samples, the same chemical forms were identified but in various ratios. Cu and Zn occurred in several chemical forms, with typically 5-7 forms present in the same sample. For Cu, the XANES spectra of the fly ash samples were nearly identical, indicating very similar chemical speciation (same chemical forms and similar ratios). Cu was found to exist in various oxide, hydroxide, chloride, silicate, and metallic forms. The most commonly occurring Zn compounds were the aluminate, ferrite, silicate, and oxide along with chloride, basic carbonate (hydrozincite), and occasionally metallic forms, probably alloyed with Cu in brass. Cu occurred in different oxidation states from zero to +II, with a higher prevalence of the lower oxidation states in bottom ash than in fly ash. Zn occurred mainly in oxidation state +II in all ashes analyzed. Finally, we showed that during outdoor storage of bottom ash, levels of Cu and Zn hydroxycarbonates were increased compared to fresh bottom ash. This carbonate formation aims to make Cu and Zn less leachable.

Bruton's tyrosine kinase (BTK) is a key component in the B-cell receptor signaling pathway and is consequently a target for in vivo imaging of B-cell malignancies as well as in multiple sclerosis (MS) with positron emission tomography (PET). A recent Phase 2b study with Sanofi's BTK inhibitor, Tolebrutinib (also known as [a.k.a.] SAR442168, PRN2246, or BTK'168) showed significantly reduced disease activity associated with MS. Herein, we report the radiosynthesis of [11 C]Tolebrutinib ([11 C]5) as a potential PET imaging agent for BTK. The N-[11 C]acrylamide moiety of [11 C]5 was labeled by 11 C-carbonylation starting from [11 C]CO, iodoethylene, and the secondary amine precursor via a novel palladium-NiXantphos-mediated carbonylation protocol, and the synthesis was fully automated using a commercial carbon-11 synthesis platform (TracerMaker™, Scansys Laboratorieteknik). [11 C]5 was obtained in a decay-corrected radiochemical yield of 37 ± 2% (n = 5, relative to starting [11 C]CO activity) in >99% radiochemical purity, with an average molar activity of 45 GBq/μmol (1200 mCi/μmol). We envision that this methodology will be generally applicable for the syntheses of labeled N-acrylamides.

The mechanism of aging is not yet fully understood. It has been recognized that there are age-dependent changes in the DNA methylation pattern of the whole genome. To date, there are several DNA methylation-based estimators of the chronological age. A majority of the estimators use the DNA methylation data from a single tissue type, such as blood. In 2013, for the first time, Steve Horvath reported the DNA methylation-based age estimator (353 CpGs were used) that could be applied to multiple tissues. A refined, more sensitive version that uses 391 CpGs was subsequently developed and validated in human cells, including fibroblasts. In this review, the age predicted by DNA methylation-based age estimator is referred to as DNAmAge, and the biological process controlling the progression of DNAmAge is referred to as the epigenetic aging in this minireview. The concepts of DNAmAge and epigenetic aging provide us opportunities to discover previously unrecognized biological events controlling aging. In this article, we discuss the frequently asked questions regarding DNAmAge and the epigenetic aging by introducing recent studies of ours and others. We focus on addressing the following questions: (1) Is there any synchronization of DNAmAge between cells in a human body?, (2) Can we use in vitro (cell culture) systems to study the epigenetic aging?, (3) Is there an age limit of DNAmAge?, and (4) Is it possible to change the speed and direction of the epigenetic aging? We describe our current understandings to these questions and outline potential future directions. Impact statement Aging is associated with DNA methylation (DNAm) changes. Recent advancement of the whole-genome DNAm analysis technology allowed scientists to develop DNAm-based age estimators. A majority of these estimators use DNAm data from a single tissue type such as blood. In 2013, a multi-tissue age estimator using DNAm pattern of 353 CpGs was developed by Steve Horvath. This estimator was named “epigenetic clock”, and the improved version using DNAm pattern of 391 CpGs was developed in 2018. The estimated age by epigenetic clock is named DNAmAge. DNAmAge can be used as a biomarker of aging predicting the risk of age-associated diseases and mortality. Although the DNAm-based age estimators were developed, the mechanism of epigenetic aging is still enigmatic. The biological significance of epigenetic aging is not well understood, either. This minireview discusses the current understanding of the mechanism of epigenetic aging and the future direction of aging research.

Energy transfer (FRET) proceeds in orthogonal dyads in contrast to Forster's theory and cannot be prohibited even by rigid interconnecting cage compounds such as cubane or diamantane, respectively.

Sperm-mediated gene transfer (SMGT) has a potential use for zebrafish transgenesis. However, transfection into fish sperm cells still needs to be improved. The objective was to demonstrate the feasibility of tip type electroporation in zebrafish sperm, showing a protocol that provide high transfection efficiency, with minimal side-effects. Sperm was transfected with a Cy3-labelled DNA using tip type electroporation with voltages ranging from 500 to 1500 V. Sperm kinetics parameters were assessed using Computer Assisted Semen Analysis (CASA) and cell integrity, reactive oxygen species (ROS), mitochondrial functionality and transfection rate were evaluated by flow cytometry. The transfection rates were positively affected by tip type electroporation, reaching 64.9% ± 3.6 in the lowest voltage used (500 V) and 86.6% ± 1.9 in the highest (1500 V). The percentage of overall motile sperm in the electrotransfected samples was found to decrease with increasing field strength (P 0.05). Overall results indicate that tip type electroporation enhances the internalization of exogenous DNA into zebrafish sperm cells with minimal harmful effects to sperm cells. Electronic supplementary material The online version of this article (10.1007/s11033-020-05658-2) contains supplementary material, which is available to authorized users.

ObjectiveThe rs641738C>T variant located near the membrane-bound O-acyltransferase domain containing 7 (MBOAT7) locus is associated with fibrosis in liver diseases, including non-alcoholic fatty liver disease (NAFLD), alcohol-related liver disease, hepatitis B and C. We aim to understand the mechanism by which the rs641738C>T variant contributes to pathogenesis of NAFLD.DesignMice with hepatocyte-specific deletion of MBOAT7 (Mboat7Δhep) were generated and livers were characterised by histology, flow cytometry, qPCR, RNA sequencing and lipidomics. We analysed the association of rs641738C>T genotype with liver inflammation and fibrosis in 846 NAFLD patients and obtained genotype-specific liver lipidomes from 280 human biopsies.ResultsAllelic imbalance analysis of heterozygous human liver samples pointed to lower expression of the MBOAT7 transcript on the rs641738C>T haplotype. Mboat7Δhep mice showed spontaneous steatosis characterised by increased hepatic cholesterol ester content after 10 weeks. After 6 weeks on a high fat, methionine-low, choline-deficient diet, mice developed increased hepatic fibrosis as measured by picrosirius staining (p<0.05), hydroxyproline content (p<0.05) and transcriptomics, while the inflammatory cell populations and inflammatory mediators were minimally affected. In a human biopsied NAFLD cohort, MBOAT7 rs641738C>T was associated with fibrosis (p=0.004) independent of the presence of histological inflammation. Liver lipidomes of Mboat7Δhep mice and human rs641738TT carriers with fibrosis showed increased total lysophosphatidylinositol levels. The altered lysophosphatidylinositol and phosphatidylinositol subspecies in MBOAT7Δhep livers and human rs641738TT carriers were similar.ConclusionMboat7 deficiency in mice and human points to an inflammation-independent pathway of liver fibrosis that may be mediated by lipid signalling and a potentially targetable treatment option in NAFLD.

Aims The aims of this study were to investigate the correlation and sex differences between total valve calcium, valve calcium concentration, and aortic valve calcification (AVC) in explanted valves from patients with severe aortic valve stenosis undergoing aortic valve replacement (AVR). Methods and results Sixty-nine patients with severe aortic stenosis (AS) scheduled for elective AVR underwent echocardiography and cardiac computed tomography (CT) prior to surgery (AVCin vivo) and CT of the explanted aortic valve (AVCex vivo). Explanted valves were prepared in acid solution, sonicated, and analysed with Arsenazo III dye to estimate total valve calcium and valve calcium concentration. Median AVCex vivo was 2082 (1421–2973) AU; mean valve calcium concentration was 1.43 ± 0.42 µmol Ca2+/mg tissue; median total valve calcium 156 (111–255) mg Ca2+, and valve calcium density 52 (35–81) mg/cm2. AVC displayed a strong correlation with total valve calcium (R2 = 0.98, P Conclusion AVC score provides a strong estimate for total valve calcium but to a lesser degree calcium concentration in the valve tissue of patients with severe AS. Females presented lower valvular calcium concentrations than males irrespective of AS severity, adding evidence and providing support to the important point that sex differences in valvular calcium concentration in AS does not reflect valvular size.