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Two experiments were conducted in which glass eels, Anguilla anguilla L., were fed with six different diets. Two natural diets, cod roe and minced bovine spleen, and four artificial diets, an unflavoured trout fry crumble and three trout fry crumble diets with added feeding stimulants, were used. The feeding stimulants tested were cod roe extract, spleen extract and a mixture of synthetic L-amino acids. In the first experiment the glass eels were fed at a restricted feeding level and in the second experiment at an ad libitum feeding level. The intake of metabolizable energy (ME) was determined as the sum of retained energy and heat production. Metabolizability of the diets decreased with increasing feeding level and was 75–95% for cod roe, 35–50% for spleen and 40–60% for the artificial diets. Since utilization of ME for maintenance and growth was not affected by feeding level and diet, differences in growth rates could be explained by differences in ME intake. Group ME intake in turn was determined by the fraction of the fish which were actually eating. The uniform feeding response of the glass eels fed the natural diets resulted in the highest ad libitum ME intake (80–90 kJ/kg0.8/day) and growth rate (4.4–4.9 g/kg0.8/day). Addition of feeding stimulants significantly increased the ad libitum ME intake of the artificial diets from 45 kJ/kg0.8/day to 50–60 kJ/kg0.8/day, which resulted in an increase in growth rate from 1.3 g/kg0.8/day to 2.4 g/kg0.8/day. [ABSTRACT FROM AUTHOR]

Abstract Background Multiple studies have shown that brain gene expression is disturbed in subjects suffering from schizophrenia. However, disentangling disease effects from alterations caused by medication is a challenging task. The main goal of this study is to find transcriptional alterations in schizophrenia that are independent of neuroleptic treatment. Methods We compared the transcriptional profiles in brain autopsy samples from 55 control individuals with that from 55 schizophrenic subjects, subdivided according to the type of antipsychotic medication received. Results Using global and high-resolution mRNA quantification techniques, we show that genes involved in immune response (GO:0006955) are up regulated in all groups of patients, including those not treated at the time of death. In particular, IFITM2, IFITM3, SERPINA3, and GBP1 showed increased mRNA levels in schizophrenia (p-values from qPCR ≤ 0.01). These four genes were co-expressed in both schizophrenic subjects and controls. In-vitro experiments suggest that these genes are expressed in both oligodendrocyte and endothelial cells, where transcription is inducible by the inflammatory cytokines TNF-α, IFN-α and IFN-γ. Conclusion Although the modified genes are not classical indicators of chronic or acute inflammation, our results indicate alterations of inflammation-related pathways in schizophrenia. In addition, the observation in oligodendrocyte cells suggests that alterations in inflammatory-related genes may have consequences for myelination. Our findings encourage future research to explore whether anti-inflammatory agents can be used in combination with traditional antipsychotics for a more efficient treatment of schizophrenia.

To protect human health, wildlife and the aquatic environment, "safe uses" of pesticides are determined at the EU level while product authorization and terms of use are established at the national level. In Sweden, extra precaution is taken to protect drinking water, and permits are therefore required for pesticide use within abstraction zones. This paper presents MACRO-DB, a tool for assessing pesticide contamination risks of groundwater and surface water, used by authorities to support their decision-making for issuing such permits. MACRO-DB is a meta-model based on 583,200 simulations of the physically-based MACRO model used for assessing pesticide leaching risks at EU and national level. MACRO-DB is simple to use and runs on widely available input data. In a qualitative comparative assessment for two counties in Sweden, MACRO-DB outputs were in general agreement with groundwater monitoring data and matched or were more protective than the national risk assessment procedure for groundwater., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Novel tumor-on-a-chip approaches are increasingly used to investigate tumor progression and potential treatment options. To improve the effect of any cancer treatment it is important to have an in depth understanding of drug diffusion, penetration through the tumor extracellular matrix and cellular uptake. In this study, we have developed a miniaturized chip where drug diffusion and cellular uptake in different hydrogel environments can be quantified at high resolution using live imaging. Diffusion of doxorubicin was reduced in a biomimetic hydrogel mimicking tissue properties of cirrhotic liver and early stage hepatocellular carcinoma (373 ± 108 µm 2 /s) as compared to an agarose gel (501 ± 77 µm 2 /s, p = 0.019). The diffusion was further lowered to 256 ± 30 µm 2 /s (p = 0.028) by preparing the biomimetic gel in cell media instead of phosphate buffered saline. The addition of liver tumor cells (Huh7 or HepG2) to the gel, at two different densities, did not significantly influence drug diffusion. Clinically relevant and quantifiable doxorubicin concentration gradients (1-20 µM) were established in the chip within one hour. Intracellular increases in doxorubicin fluorescence correlated with decreasing fluorescence of the DNA-binding stain Hoechst 33342 and based on the quantified intracellular uptake of doxorubicin an apparent cell permeability (9.00 ± 0.74 × 10 -4  µm/s for HepG2) was determined. Finally, the data derived from the in vitro model were applied to a spatio-temporal tissue concentration model to evaluate the potential clinical impact of a cirrhotic extracellular matrix on doxorubicin diffusion and tumor cell uptake., (© 2023. The Author(s).)

Silicon nitride (SiN) coatings may reduce unwanted release of metal ions from metallic implants. However, as SiN slowly dissolves in aqueous solutions, additives that reduce this dissolution rate would likely increase the lifetime and functionality of implants. Adding iron (Fe) and carbon (C) permits tuning of the SiN coatings' mechanical properties, but their effect on SiN dissolution rates, and their capacity to reduce metal ion release from metallic implant substrates, have yet to be investigated. Such coatings have recently been proposed for use in spinal implants; therefore, it is relevant to assess their impact on the viability of cells expected at the implant site, such as microglia, the resident macrophages of the central nervous system (CNS). To study the effects of Fe and C on the dissolution rate of SiN coatings, compositional gradients of Si, Fe and C in combination with N were generated by physical vapor deposition onto CoCrMo discs. Differences in composition did not affect the surface roughness or the release of Si, Fe or Co ions (the latter from the CoCrMo substrate). Adding Fe and C reduced ion release compared to a SiN reference coating, which was attributed to altered reactivity due to an increase in the fraction of stabilizing Si-C or Fe-C bonds. Extracts from the SiN coatings containing Fe and C were compatible with microglial viability in 2D cultures and 3D collagen hydrogels, to a similar degree as CoCrMo and SiN coated CoCrMo reference extracts. As Fe and C reduced the dissolution rate of SiN-coatings and did not compromise microglial viability, the capacity of these additives to extend the lifetime and functionality of SiN-coated metallic implants warrants further investigation.

The primary cilium is an organelle present in most adult mammalian cells that is considered as an antenna for sensing the local microenvironment. Here, we use intact mouse pancreatic islets of Langerhans to investigate signaling properties of the primary cilium in insulin-secreting β-cells. We find that GABAB1 receptors are strongly enriched at the base of the cilium, but are mobilized to more distal locations upon agonist binding. Using cilia-targeted Ca2+ indicators, we find that activation of GABAB1 receptors induces selective Ca2+ influx into primary cilia through a mechanism that requires voltage-dependent Ca2+ channel activation. Islet β-cells utilize cytosolic Ca2+ increases as the main trigger for insulin secretion, yet we find that increases in cytosolic Ca2+ fail to propagate into the cilium, and that this isolation is largely due to enhanced Ca2+ extrusion in the cilium. Our work reveals local GABA action on primary cilia that involves Ca2+ influx and depends on restricted Ca2+ diffusion between the cilium and cytosol., (© 2022 Sanchez et al.)

Biofilms are arguably the most important mode of growth of bacteria, but how antibiotic resistance emerges and is selected in biofilms remains poorly understood. Several models to study evolution of antibiotic resistance have been developed, however, their usability varies depending on the nature of the biological question. Here, we developed and validated a microfluidic chip (Brimor) for studying the dynamics of enrichment of antibiotic-resistant bacteria in biofilms using real-time monitoring with confocal microscopy. In situ extracellular cellulose staining and physical disruption of the biomass confirmed Escherichia coli growth as biofilms in the chip. We showed that seven generations of growth occur in 16 h when biofilms were established in the growth chambers of Brimor, and that bacterial death and growth rates could be estimated under these conditions using a plasmid with a conditional replication origin. Additionally, competition experiments between antibiotic-susceptible and -resistant bacteria at sub-inhibitory concentrations demonstrated that the antibiotic ciprofloxacin selected for antibiotic resistance in bacterial biofilms at concentrations 17-fold below the minimal inhibitory concentration of susceptible planktonic bacteria. Overall, the microfluidic chip is easy to use and a relevant model for studying the dynamics of selection of antibiotic resistance in bacterial biofilms and we anticipate that the Brimor chip will facilitate basic research in this area., Competing Interests: A patent application comprising the chip design and the here described method has been filed by P-CT, OE, and JK under number PCT/SE2019/050635. JS is currently employed by Corline Biomedical AB. The company had no part in the design, performance, or analysis of data in the study. The remaining author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Tang, Eriksson, Sjögren, Fatsis-Kavalopoulos, Kreuger and Andersson.)

Filter-aided sample preparation (FASP) is widely used in bottom-up proteomics for tryptic digestion. However, the sample recovery yield of this method is limited by the amount of the starting material. While ∼100 ng of digested protein is sufficient for thorough protein identification, proteomic information gets lost with a protein content <10 μg due to incomplete peptide recovery from the filter. We developed and optimized a flexible well-plate μFASP device and protocol that is suitable for an ∼1 μg protein sample. In 1 μg of HeLa digest, we identified 1295 ± 10 proteins with μFASP followed by analysis with liquid chromatography-mass spectrometry. In contrast, only 524 ± 5 proteins were identified with the standard FASP protocol, while 1395 ± 4 proteins were identified in 20 μg after standard FASP as a benchmark. Furthermore, we conducted a combined peptidomic and proteomic study of single pancreatic islets with well-plate μFASP. Here, we separated neuropeptides and digested the remaining on-filter proteins for bottom-up proteomic analysis. Our results indicate inter-islet heterogeneity for the expression of proteins involved in glucose catabolism, pancreatic hormone processing, and secreted peptide hormones. We consider our method to provide a useful tool for proteomic characterization of samples where the biological material is scarce. All proteomic data are available under DOI: 10.6019/PXD029039.

Cancer cells exploit a variety of migration modes to leave primary tumors and establish metastases, including amoeboid cell migration, which is typically reliant on bleb formation. Here we demonstrate that thrombin induces dynamic blebbing in the MDA-MB-231 breast cancer cell line and confirm that protease-activated receptor 1 (PAR1) activation is sufficient to induce this effect. Cell confinement has been implicated as a driving force in bleb-based migration. Unexpectedly, we found that gentle contact compression, exerted using a custom built 'cell press' to mechanically stimulate cells, reduced thrombin-induced blebbing. Thrombin-induced blebbing was similarly attenuated using the small molecule Yoda1, an agonist of the mechanosensitive Ca2+ channel Piezo1, and this attenuation was impaired in Piezo1-depleted cells. Additionally, Piezo1 activation suppressed thrombin-induced phosphorylation of ezrin, radixin and moesin (ERM) proteins, which are implicated in the blebbing process. Our results provide mechanistic insights into Piezo1 activation as a suppressor of dynamic blebbing, specifically that which is induced by thrombin., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2022. Published by The Company of Biologists Ltd.)

The rapidly changing landscape of antimicrobial resistance poses a challenge for empirical antibiotic therapy in severely ill patients and highlights the need for fast antibiotic susceptibility diagnostics to guide treatment. Traditional methods for antibiotic susceptibility testing (AST) of bacteria such as broth microdilution (BMD) or the disc diffusion method (DDM) are comparatively slow and show high variability. Rapid AST methods under development often trade speed for resolution, sometimes only measuring responses at a single antibiotic concentration. QuickMIC is a recently developed lab-on-a-chip system for rapid AST. Here we evaluate the performance of the QuickMIC method with regard to speed, precision and accuracy in comparison to traditional diagnostic methods. 151 blood cultures of clinical Gram-negative isolates with a high frequency of drug resistance were tested using the QuickMIC system and compared with BMD for 12 antibiotics. To investigate sample turnaround time and method functionality in a clinical setting, another 41 clinical blood culture samples were acquired from the Uppsala University Hospital and analyzed on site in the clinical laboratory with the QuickMIC system, and compared with DDM for 8 antibiotics routinely used in the clinical laboratory. The overall essential agreement between MIC values obtained by QuickMIC and BMD was 83.4%, with an average time to result of 3 h 2 min (SD: 24.8 min) for the QuickMIC method. For the clinical dataset, the categorical agreement between QuickMIC and DDM was 96.8%, whereas essential and categorical agreement against BMD was 91.0% and 96.7%, respectively, and the total turnaround time as compared to routine diagnostics was shown to be reduced by 40% (33 h vs. 55 h). Interexperiment variability was low (average SD: 44.6% from target MIC) compared to the acceptable standard of ±1 log 2 unit ( i.e. -50% to +100% deviation from target MIC) in BMD. We conclude that the QuickMIC method can provide rapid and accurate AST, and may be especially valuable in settings with high resistance rates, and for antibiotics where wildtype and antibiotic-resistant bacteria have MIC distributions that are close or overlapping., Competing Interests: CM is a part-time employee of Gradientech AB. JT, JF, HÖ, CJ and JÅ are employees of Gradientech AB. JK is not employed by Gradientech AB but is a co-founder of Gradientech AB and owns stock in the company. The remaining author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Malmberg, Torpner, Fernberg, Öhrn, Ångström, Johansson, Tängdén and Kreuger.)

Greenhouse and other covered cultivation systems have increased globally over the past several decades, leading to considerably improved product quality and productivity per land area unit. However, there is a paucity in information regarding the environmental impacts of covered production systems, especially regarding pesticides entering the surrounding environment. Aiming to address this knowledge gap, we collected grab samples downstream of greenhouses from seven Swedish streams every 14 days during a 12 month period. In three of the streams, samples were also taken upstream of the greenhouses and in four of the streams time-integrated samples were collected by TIMFIE samplers in the period between grab sampling occasions. The samples were analyzed for 28 substances (27 that were permitted for use in greenhouse production systems in Sweden and one degradation product to a permitted substance). Pesticide use journals were collected from the greenhouse producers for the 12 month period. The results were examined for indications of greenhouse contributions to detection frequencies, maximum and average concentrations, and potential ecotoxicicity in several ways: (1) comparing locations downstream of greenhouses with registered use of a substance with those without registered use, (2) comparing results from this study with those from the Swedish environmental monitoring program of pesticides in surface water from catchments with no greenhouses from the same period and region, (3) comparing concentration trends with registered pesticide application times in the greenhouses, and (4) comparing up- and downstream concentrations. The results strongly suggest that greenhouse applications do contribute to pesticide occurrences, maximum and median concentrations for most of the pesticides included in this study, and to potential toxicity to aquatic organisms for several of them, most notably imidacloprid, acetamiprid, carbendazim, and pirimicarb., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)

Bioprinting is increasingly used to create complex tissue constructs for an array of research applications, and there are also increasing efforts to print tissues for transplantation. Bioprinting may also prove valuable in the context of drug screening for personalized medicine for treatment of diseases such as cancer. However, the rapidly expanding bioprinting research field is currently limited by access to bioprinters. To increase the availability of bioprinting technologies we present here an open source extrusion bioprinter based on the E3D motion system and tool changer to enable high-resolution multimaterial bioprinting. As proof of concept, the bioprinter is used to create collagen constructs using freeform reversible embedding of suspended hydrogels (FRESH) methodology, as well as multimaterial constructs composed of distinct sections of laminin and collagen. Data is presented demonstrating that the bioprinted constructs support growth of cells either seeded onto printed constructs or included in the bioink prior to bioprinting. This open source bioprinter is easily adapted for different bioprinting applications, and additional tools can be incorporated to increase the capabilities of the system., (© 2021. The Author(s).)

Pesticides are widespread anthropogenic chemicals and well-known environmental contaminants of concern. Much less is known about transformation products (TPs) of pesticides and their presence in the environment. We developed a novel suspect screening approach for not well-explored pesticides ( n = 16) and pesticide TPs ( n = 242) by integrating knowledge from national monitoring with high-resolution mass spectrometry data. Weekly time-integrated samples were collected in two Swedish agricultural streams using the novel Time-Integrating, MicroFlow, In-line Extraction (TIMFIE) sampler. The integration of national monitoring data in the screening approach increased the number of prioritized compounds approximately twofold (from 23 to 42). Ultimately, 11 pesticide TPs were confirmed by reference standards and 12 TPs were considered tentatively identified with varying levels of confidence. Semiquantification of the newly confirmed TPs indicated higher concentrations than their corresponding parent pesticides in some cases, which highlights concerns related to (unknown) pesticide TPs in the environment. Some TPs were present in the environment without co-occurrence of their corresponding parent compounds, indicating higher persistency or mobility of the identified TPs. This study showcased the benefits of integrating monitoring knowledge in this type of studies, with advantages for suspect screening performance and the possibility to increase relevance of future monitoring programs.

Synthetic bone models are used to train surgeons as well as to test new medical devices. However, currently available models do not accurately mimic the complex structure of trabecular bone, which can provide erroneous results. This study aimed to investigate the suitability of stereolithography (SLA) to produce synthetic trabecular bone. Samples were printed based on synchrotron micro-computed tomography (micro-CT) images of human bone, with scaling factors from 1 to 4.3. Structure replicability was assessed with micro-CT, and mechanical properties were evaluated by compression and screw pull-out tests. The overall geometry was well-replicated at scale 1.8, with a volume difference to the original model of <10%. However, scaling factors below 1.8 gave major print artefacts, and a low accuracy in trabecular thickness distribution. A comparison of the model-print overlap showed printing inaccuracies of ~20% for the 1.8 scale, visible as a loss of smaller details. SLA-printed parts exhibited a higher pull-out strength compared to existing synthetic models (Sawbones ™), and a lower strength compared to cadaveric specimens and fused deposition modelling (FDM)-printed parts in poly (lactic acid). In conclusion, for the same 3D model, SLA enabled higher resolution and printing of smaller scales compared to results reported by FDM.

Long-term pesticide water concentrations were investigated in four agricultural streams and their mixture toxicity on algae was assessed, based on realistic (i.e. observed) concentrations in laboratory tests using (i) natural weekly water samples and (ii) reconstituted pesticide-spiked water samples representing mixtures with predicted high mixture. This approach both covered the full complexity of natural water samples and the controlled approach of reconstituted water samples. Long-term monitoring data (time-integrated, weekly samples) revealed more than 11 pesticides (range 11.0 ± 0.25-24.0 ± 0.44) in 75% or more of the almost 1600 samples collected between 2002 and 2018. ∑TU algae exceeded 0.1 for 29 observations (or 1.8%). Despite the multitude of pesticides in a sample, ∑TU algae was frequently set by one or a few dominating pesticides that contribute to more than 90% of the mixture's toxicity. Algal growth inhibition tests with in situ stream water showed a high frequency of inhibition, despite the low ∑TU for most of these samples (range 0.000014-0.3858). These "false positive" results were attributed to confounding effects of turbidity, the complexation of nutrients, and toxic effects of metals and/or other unknown contaminants. Algal inhibition tests with spiked reconstituted water showed significant inhibitory effects in the range of 1-10x the ∑TU algae observed in worst-case field samples. Although these tests disregard the chemical complexity of natural water, they show that inhibitory effects of pesticides on algae may occur at the ∑TU algae observed in monitoring. Furthermore, considering that the ∑TU algae of stream water are based on weekly average concentrations and likely underestimate short-term peak concentrations of pesticides, these results strongly suggest that inhibitory effects on algae may occur in the agricultural streams of southern Sweden. We conjecture, however, that the rapid recovery of algae contributes to ameliorate these short-term effects and that pesticide contamination should be seen as one of many stressors in the streams that drain agricultural landscapes., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Antibiotic combination therapies are important for the efficient treatment of many types of infections, including those caused by antibiotic-resistant pathogens. Combination treatment strategies are typically used under the assumption that synergies are conserved across species and strains, even though recent results show that the combined treatment effect is determined by specific drug-strain interactions that can vary extensively and unpredictably, both between and within bacterial species. To address this problem, we present a new method in which antibiotic synergy is rapidly quantified on a case-by-case basis, allowing for improved combination therapy. The novel CombiANT methodology consists of a 3D-printed agar plate insert that produces defined diffusion landscapes of 3 antibiotics, permitting synergy quantification between all 3 antibiotic pairs with a single test. Automated image analysis yields fractional inhibitory concentration indices (FICis) with high accuracy and precision. A technical validation with 3 major pathogens, Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus, showed equivalent performance to checkerboard methodology, with the advantage of strongly reduced assay complexity and costs for CombiANT. A synergy screening of 10 antibiotic combinations for 12 E. coli urinary tract infection (UTI) clinical isolates illustrates the need for refined combination treatment strategies. For example, combinations of trimethoprim (TMP) + nitrofurantoin (NIT) and TMP + mecillinam (MEC) showed synergy, but only for certain individual isolates, whereas MEC + NIT combinations showed antagonistic interactions across all tested strains. These data suggest that the CombiANT methodology could allow personalized clinical synergy testing and large-scale screening. We anticipate that CombiANT will greatly facilitate clinical and basic research of antibiotic synergy., Competing Interests: The authors of this manuscript have the following competing interests: A patent comprising the described method has been filed by the authors and is pending under number 640 SE 2050304-1.