Publications - Publications en-us PURE Extension (Web Department) 30 <![CDATA[Preoperative BRAF<sup>V600E</sup> mutation detection in thyroid carcinoma by immunocytochemistry]]> Swan, K. Z., Madsen, S. H., Bonnema, S. J., Nielsen, V. E., Jespersen, M. L. The BRAFV600E (BRAF) mutation is present in 40–50% of papillary thyroid carcinomas (PTC) and has been associated with more aggressive clinicopathological characteristics of PTC. The aim of this study was to evaluate different methods for preoperative identification of the BRAF mutation in PTC using cytological and histological specimens. Prospectively collected preoperative cytological clots from patients with suspected PTC were tested with BRAF immunocytochemistry (ICC) and the Cobas Test (PCR). In addition, histological specimens were tested with BRAF immunohistochemistry (IHC) and the Cobas Test. All nodules were histologically examined. Fifty-three patients were included in the study. Complete mutation testing was available in 32 patients. The main reason for exclusion was insufficient cell content in the cytological specimen. Twenty-seven nodules were histologically diagnosed as PTC, and 41% (n = 11) of PTCs were BRAF ICC positive. All non-PTC nodules were negative by BRAF ICC. In 26 nodules, all four BRAF tests were concordant, while discordant test results were found in six nodules. ICC was in accordance with the consensus BRAF status in five of these nodules, while BRAF status was undetermined in one nodule. BRAF ICC showed high concordance with the Cobas Test and a low rate of false negative stain. These results indicate that BRAF ICC may be a feasible method for preoperative detection of the BRAFV600E mutation in patients with PTC.

Research Tue, 01 Nov 2022 17:51:39 +0100 32ae0dca-756e-4a6c-8d0b-83bdd9739d6f
<![CDATA[Spatial Organization of Osteoclastic Coupling Factors and Their Receptors at Human Bone Remodeling Sites]]> Borggaard, X. G., Nielsen, M. H., Delaisse, J. M., Andreasen, C. M., Andersen, T. L. The strictly regulated bone remodeling process ensures that osteoblastic bone formation is coupled to osteoclastic bone resorption. This coupling is regulated by a panel of coupling factors, including clastokines promoting the recruitment, expansion, and differentiation of osteoprogenitor cells within the eroded cavity. The osteoprogenitor cells on eroded surfaces are called reversal cells. They are intermixed with osteoclasts and become bone-forming osteoblast when reaching a critical density and maturity. Several coupling factors have been proposed in the literature, but their effects and expression pattern vary between studies depending on species and experimental setup. In this study, we investigated the mRNA levels of proposed secreted and membrane-bound coupling factors and their receptors in cortical bone remodeling events within the femur of healthy adolescent human controls using high-sensitivity RNA in situ hybridization. Of the proposed coupling factors, human osteoclasts showed mRNA-presence of LIF, PDGFB, SEMA4D, but no presence of EFNB2, and OSM. On the other hand, the osteoblastic reversal cells proximate to osteoclasts presented with LIFR, PDGFRA and PLXNB1, but not PDGFRB, which are all known receptors of the proposed coupling factors. Although EFNB2 was not present in mature osteoclasts, the mRNA of the ligand-receptor pair EFNB2:EPHB4 were abundant near the central blood vessels within intracortical pores with active remodeling. EPHB4 and SEMA4D were also abundant in mature bone-forming osteoblasts. This study highlights that especially LIF:LIFR, PDGFB:PDGFRA, SEMA4D:PLXNB1 may play a critical role in the osteoclast-osteoblast coupling in human remodeling events, as they are expressed within the critical cells.

Research Wed, 01 Jun 2022 17:51:39 +0200 bb734f76-4b4a-4284-a7e2-7147d9756c3c
<![CDATA[Senicapoc treatment in COVID-19 patients with severe respiratory insufficiency]]> Granfeldt, A., Andersen, L. W., Fink Vallentin, M., et al. Background: The aim of the current study was to determine if treatment with senicapoc, improves the PaO2/FiO2 ratio in patients with COVID-19 and severe respiratory insufficiency. Methods: Investigator-initiated, randomized, open-label, phase II trial in four intensive care units (ICU) in Denmark. We included patients aged ≥18 years and admitted to an ICU with severe respiratory insufficiency due to COVID-19. The intervention consisted of 50 mg enteral senicapoc administered as soon as possible after randomization and again after 24 h. Patients in the control group received standard care only. The primary outcome was the PaO2/FiO2 ratio at 72 h. Results: Twenty patients were randomized to senicapoc and 26 patients to standard care. Important differences existed in patient characteristics at baseline, including more patients being on non-invasive/invasive ventilation in the control group (54% vs. 35%). The median senicapoc concentration at 72 h was 62.1 ng/ml (IQR 46.7–71.2). The primary outcome, PaO2/FiO2 ratio at 72 h, was significantly lower in the senicapoc group (mean 19.5 kPa, SD 6.6) than in the control group (mean 24.4 kPa, SD 9.2) (mean difference −5.1 kPa [95% CI −10.2, −0.04] p =.05). The 28-day mortality in the senicapoc group was 2/20 (10%) compared with 6/26 (23%) in the control group (OR 0.36 95% CI 0.06–2.07, p =.26). Conclusions: Treatment with senicapoc resulted in a significantly lower PaO2/FiO2 ratio at 72 h with no differences for other outcomes.

Research Mon, 01 Aug 2022 17:51:39 +0200 193ffe39-0a48-4c84-b88d-0ec56cfaaef8
<![CDATA[Investigating effects of sodium beta-hydroxybutyrate on metabolism in placebo-controlled, bilaterally infused human leg with focus on skeletal muscle protein dynamics]]> Thomsen, H. H., Olesen, J. F., Aagaard, R., et al. Systemic administration of beta-hydroxybutyrate (BHB) decreases whole-body protein oxidation and muscle protein breakdown in humans. We aimed to determine any direct effect of BHB on skeletal muscle protein turnover when administered locally in the femoral artery. Paired design with each subject being investigated on one single occasion with one leg being infused with BHB and the opposing leg acting as a control. We studied 10 healthy male volunteers once with bilateral femoral vein and artery catheters. One artery was perfused with saline (Placebo) and one with sodium-BHB. Labelled phenylalanine and palmitate were used to assess local leg fluxes. Femoral vein concentrations of BHB were significantly higher in the intervention leg (3.4 (3.2, 3.6) mM) compared with the placebo-controlled leg (1.9 (1.8, 2.1) mM) with a peak difference of 1.4 (1.1, 1.7) mM, p < 0.0005. Net loss of phenylalanine for BHB vs Placebo -6.7(-10.8, -2.7) nmol/min vs -8.7(-13.8, -3.7) nmol/min, p = 0.52. Palmitate flux and arterio-venous difference of glucose did not differ between legs. Under these experimental conditions, we failed to observe the direct effects of BHB on skeletal muscle protein turnover. This may relate to a combination of high concentrations of BHB (close to 2 mM) imposed systemically by spillover leading to high BHB concentrations in the saline-infused leg and a lack of major differences in concentration gradients between the two sides-implying that observations were made on the upper part of the dose-response curve for BHB and the relatively small number of subjects studied.

Research Mon, 01 Aug 2022 17:51:39 +0200 df7bc4d8-e18b-4321-a326-af41e5d6ede8
<![CDATA[Blunt force homicides in Denmark 1992-2016]]> Thomsen, A. H., Leth, P. M., Hougen, H. P., Villesen, P. Blunt force trauma is a common homicide method, inflicted in three different ways: bodily force, assault with blunt objects of various types and falls from height. The objective of this study is to provide thorough information on blunt force homicides with data on the victims, the offenders, the surrounding circumstances, the injury methods, the extent of injuries, and survival time, which will help inform the inexperienced as well as the seasoned forensic pathologist in their daily work with death investigation and as expert witnesses in court. We have analyzed autopsy reports and available case files of 311 blunt force homicides, making up 21.9% of all homicides in Denmark during 1992-2016. Most victims and offenders were male. Altercation in the setting of nightlife and intoxication was common in male victims, while most female victims were killed in a domestic setting. Bodily force was the most common primary homicide method, followed by assault with a blunt object and fall from height. The head was the region that most often had external injuries, with no noteworthy difference between cases with bodily force and blunt objects. Two out of three victims had one or more lacerations, most often located on the head and more often on the front. Brain injury was the primary cause of death in at least 72.0% victims. Compared to bodily force victims of blunt object assault were especially prone to skull and brain injuries, had a higher trauma score, and more died at the crime scene and had a shorter survival time.

Research Tue, 01 Nov 2022 17:51:39 +0100 60f7640a-37db-43b6-b550-f3c891e39de0
<![CDATA[Efficacy of Chronic Paroxetine Treatment in Mitigating Amyloid Pathology and Microgliosis in APP<sub>SWE/PS1</sub>ΔE9 Transgenic Mice]]> Sivasaravanaparan, M., Olesen, L. Ø., Severino, M., et al. Background: Modulation of serotonergic signaling by treatment with selective serotonin reuptake inhibitors (SSRIs) has been suggested to mitigate amyloid-β (Aβ) pathology in Alzheimer's disease, in addition to exerting an anti-depressant action. Objective: To investigate the efficacy of chronic treatment with the SSRI paroxetine, in mitigating Aβ pathology and Aβ plaque-induced microgliosis in the hippocampus of 18-month-old APPswe/PS1ΔE9 mice. Methods: Plaque-bearing APPswe/PS1ΔE9 and wildtype mice were treated with paroxetine per os at a dose of 5 mg/kg/day, from 9 to 18 months of age. The per os treatment was monitored by recording of the body weights and serum paroxetine concentrations, and by assessment of the serotonin transporter occupancy by [3H]DASB-binding in wildtype mice. Additionally, 5,7-dihydroxytryptamine was administered to 9-month-old APPswe/PS1ΔE9 mice, to examine the effect of serotonin depletion on Aβ pathology. Aβ pathology was evaluated by Aβ plaque load estimation and the Aβ42/Aβ40 ratio by ELISA. Results: Paroxetine treatment led to > 80% serotonin transporter occupancy. The treatment increased the body weight of wildtype mice, but not of APPswe/PS1ΔE9 mice. The treatment had no effect on the Aβ plaque load (p = 0.39), the number and size of plaques, or the Aβ plaque-induced increases in microglial numbers in the dentate gyrus. Three months of serotonin depletion did not significantly impact the Aβ plaque load or Aβ42/Aβ40 ratio in APPswe/PS1ΔE9 mice at 12 months. Conclusion: Our results show that chronic treatment with the SSRI paroxetine does not mitigate Aβ pathology and Aβ plaque-induced microgliosis in the hippocampus of APPswe/PS1ΔE9 mice.

Research Sun, 01 May 2022 17:51:39 +0200 4c7ed355-95cf-447b-a7a2-53f4e9852e7b
<![CDATA[The status of forensic radiography in the Nordic Countries]]> Primeau, C., Marttinen, F., Pedersen, C. C. E. The International Association of Forensic Radiographers (IAFR) aims to promote best practice in forensic radiography through education and training, research, communication and coordination of forensic radiography. To survey the situation of these areas a questionnaire was sent in 2020 to all institutes of forensic medicine in the Nordic countries (Denmark, Sweden, Finland, Norway and Iceland) to clarify the current status of forensic radiography. Results showed the role of forensic radiography in the Nordic countries appeared to be in the early stages of development with a distinctive lack of standardised guidelines, formalised training, an absence of national associations, and little involvement with international associations in forensic radiography. This was despite a significant amount of case work having been undertaken within forensic departments as well as within clinical settings, including work involving both deceased and living individuals. The insights gained through this questionnaire provided focus on the areas that could be targeted for further development and may facilitate strategies to promote best practice and training needs for practitioners of forensic radiography within the Nordic countries.

Research Wed, 01 Jun 2022 17:51:39 +0200 f9c7ff64-006a-4982-b390-e45cd2c69351
<![CDATA[Combined effects of quetiapine and opioids]]> Andersen, F. D., Joca, S., Hvingelby, V., et al. Research Thu, 01 Sep 2022 17:51:39 +0200 3780bc01-9364-420e-96c6-8404901a641f <![CDATA[Tager man mest amfetamin i provinsen?]]> Lindholst, C., Grasaasen, K. ]]> Communication Sat, 01 Jan 2022 17:51:39 +0100 4cb73c9d-f42e-4f6d-80e4-07322c3da4f7 <![CDATA[Steady-state concentrations of flucloxacillin in porcine vertebral cancellous bone and intervertebral disc following oral and intravenous administration assessed by microdialysis]]> Bendtsen, M. A.F., Hanberg, P., Slater, J., et al. Aims: Flucloxacillin is a frequently used antibiotic in the treatment of spondylodiscitis. We assessed steady-state concentrations and time above minimal inhibitory concentration (fT > MIC) of flucloxacillin in the intervertebral disc, vertebral cancellous bone, subcutaneous tissue and plasma, after intravenous and oral administration. Methods: Sixteen pigs were randomized into two groups; Group Peroral (Group PO) and Group Intravenous (Group IV) received 1 g flucloxacillin every 6 h for 24 h orally or intravenously. Microdialysis was used for sampling in the compartments of interest. A flucloxacillin target of 50% fT > MIC was applied for three MIC targets: 0.125, 0.5 and 2.0 μg/mL. Results: Intravenous administration resulted in significantly longer fT > MIC for all targets. Target attainment was only reached for the low target of 0.125 μg/mL in Group IV in vertebral cancellous bone, subcutaneous tissue, and plasma (intervertebral disc 47%). In Group IV, mean fT > MIC values in the investigated compartments were in the range of 47–67% of the dosing interval for 0.125 μg/mL, 20–35% for 0.5 μg/mL, and 0–15% for 2.0 μg/mL. In Group PO, mean fT > MIC values for 0.125 μg/mL were in the range of 1–33%. No pigs reached a concentration of 0.5 μg/mL in any of the investigated compartments in Group PO. Conclusion: Administration of 1 g flucloxacillin every 6 h resulted in surprisingly low steady-state fT > MIC after intravenous and oral administration. However, intravenous administration resulted in significantly higher concentrations across compartments compared to oral administration. Sufficient target tissue concentrations for treatment of spondylodiscitis may require a dose increase or alternative dosing regimens.

Research Wed, 01 Jun 2022 17:51:39 +0200 5d5e4708-7c88-4043-b8f7-97e2bc120ee7
<![CDATA[Acute drug poisonings leading to hospitalization]]> Andersen, C. U., Nielsen, L. P., Møller, J. M., Olesen, A. E. Knowledge about current trends and epidemiology in poisonings is important to maintain quality in diagnostics, treatment and prevention. We performed a cross-sectional study of all cases (n = 261) admitted with drug poisoning to Aalborg University Hospital during 1 year in 2017–2018. Median age was 30 (22–49) years, and 58% were female. Fifty percent were suicide attempts. In most cases, involved drugs were identified by history taking; blood analysis barely revealed any additional paracetamol and salicylicate poisonings. Drugs prescribed to the patient or available over the counter were involved in nearly two thirds of cases. Weak analgesics dominated by paracetamol (n = 91, 35%) was the most frequently involved group of drugs followed by opioids and benzodiazepines. Gender differences were observed with respect to involvement of weak analgesics and central stimulants. A higher prevalence of unidentified involved drugs was observed in 26 cases (10%) in which the length of admission exceeded 2 days and/or intensive care was needed. No deaths, cardiac arrhythmias or physical complications occurred. Thus, current handling of the acute poisoning seems effective in most cases. However, a more tailored use of blood analyses including a toxicological screen in selected cases may represent an opportunity for improvement.

Research Tue, 01 Feb 2022 17:51:39 +0100 85c5a388-1260-4d40-bf71-9cf576dc7c97
<![CDATA[A Retrospective Metabolomics Analysis of Gamma-Hydroxybutyrate in Humans]]> Wang, T., Nielsen, K. L., Frisch, K., et al. GHB is an endogenous short-chain organic acid presumably also widely applied as a rape and knock out drug in cases of drug-facilitated crimes or sexual assaults (DFSA). Due to the endogenous nature of GHB and its fast metabolism in vivo, the detection window of exogenous GHB is however narrow, making it challenging to prove use of GHB in DFSA cases. Alternative markers of GHB intake have recently appeared though none has hitherto been validated for forensic use. UHPLC-HRMS based screening of blood samples for drugs of abuse is routinely performed in several forensic laboratories which leaves an enormous amount of unexploited data. Recently we devised a novel metabolomics approach to use archived data from such routine screenings for elucidating both direct metabolites from exogenous compounds, but potentially also regulation of endogenous metabolism and metabolites. In this paper we used UHPLC-HRMS data acquired over a 6-year period from whole blood analysis of 51 drivers driving under the influence of GHB as well as a matched control group. The data were analyzed using a metabolomics approach applying a range of advanced analytical methods such as OPLS-DA, LASSO, random forest, and Pearson correlation to examine the data in depth and demonstrate the feasibility and potential power of the approach. This was done by initially detecting a range of potential biomarkers of GHB consumption, some that previously have been found in controlled GHB studies, as well as several new potential markers not hitherto known. Furthermore, we investigate the impact of GHB intake on human metabolism. In aggregate, we demonstrate the feasibility to extract meaningful information from archived data here exemplified using GHB cases. Hereby we hope to pave the way for more general use of the principle to elucidate human metabolites of e.g. new legal or illegal drugs as well as for applications in more global and large scale metabolomics studies in the future.

Research Tue, 01 Mar 2022 17:51:39 +0100 c7b319ca-23da-4e65-86ce-1e3c9f191771
<![CDATA[Alendronate prolongs the reversal-resorption phase in human cortical bone remodeling]]> Borggaard, X. G., Roux, J. P., Delaisse, J. M., Chavassieux, P., Andreasen, C. M., Andersen, T. L. Despite their ability to reduce fracture-risk and increase Bone Mineral Density (BMD) in osteoporotic women, bisphosphonates are reported to reduce formation of new bone. Reduced bone formation has been suggested to lead to accumulation of microfractures and contribute to rare side effects in cortical bone such as atypical femur fractures. However, most studies are limited to trabecular bone. In this study, the cortical bone remodeling in human iliac bone specimens of 65 non-treated and 24 alendronate-treated osteoporotic women was investigated using a new histomorphometric classification of intracortical pores. The study showed that only 12.4 ± 11% of the cortical pore area reflected quiescent pores/osteons in alendronate-treated patients versus 8.5 ± 5% in placebo, highlighting that new cortical remodeling events remain to be activated. The percent and size of eroded pores (events in resorption-reversal phase) remained unchanged, but their contribution to total pore area was 1.4-fold higher in alendronate versus placebo treated patients (66 ± 22% vs 48 ± 22%, p < 0.001). On the other hand, the mixed eroded-formative pores (events with mixed resorption-reversal-formation phases) was 2-fold lower in alendronate versus placebo treated patients (19 ± 14% vs 39 ± 23% of total pore area, p < 0.001), and formative pores (event in formation phase) was 2.2-fold lower in alendronate versus placebo treated patients (2.1 ± 2.4% vs 4.6 ± 3.6%, p < 0.01), and their contribution to total pore area was 2.4-fold lower (1.3 ± 2.1% vs 3.1 ± 4.4%, p < 0.05). Importantly, these differences between alendronate and placebo treated patients were significant in patients after 3 years of treatment, not after 2 years of treatment. Collectively, the results support that cortical remodeling events activated during alendronate treatment has a prolonged reversal-resorption phase with a delayed transition to formation, becoming increasingly evident after 3-years of treatment. A potential contributor to atypical femur fractures associated with long-term bisphosphonate treatment.

Research Fri, 01 Jul 2022 17:51:39 +0200 dc2116e9-4951-4d14-a167-ec04ec639c8b
<![CDATA[Hyperpolarized <sup>13</sup>C MRI Reveals Large Changes in Pyruvate Metabolism During Digestion in Snakes]]> Hansen, K., Hansen, E. S. S., Jespersen, N. R. V., et al. PURPOSE: Hyperpolarized 13C MRI is a powerful technique to study dynamic metabolic processes in vivo; but it has predominantly been used in mammals, mostly humans, pigs, and rodents.

METHODS: In the present study, we use this technique to characterize the metabolic fate of hyperpolarized [1- 13C]pyruvate in Burmese pythons (Python bivittatus), a large species of constricting snake that exhibits a four- to tenfold rise in metabolism and large growth of the visceral organs within 24-48 h of ingestion of their large meals.

RESULTS: We demonstrate a fivefold elevation of the whole-body lactate-to-pyruvate ratio in digesting snakes, pointing to a large rise in lactate production from pyruvate. Consistent with the well-known metabolic stimulation of digestion, measurements of mitochondrial respiration in hepatocytes in vitro indicate a marked postprandial upregulation of mitochondrial respiration. We observed that a low SNR of the hyperpolarized 13C produced metabolites in the python, and this lack of signal was possibly due to the low metabolism of reptiles compared with mammals, preventing quantification of alanine and bicarbonate production with the experimental setup used in this study. Spatial quantification of the [1- 13C]lactate was only possible in postprandial snakes (with high metabolism), where a statistically significant difference between the heart and liver was observed.

CONCLUSION: We confirm the large postprandial rise in the wet mass of most visceral organs, except for the heart, and demonstrated that it is possible to image the [1- 13C]pyruvate uptake and intracellular conversion to [1- 13C]lactate in ectothermic animals.

Research Mon, 01 Aug 2022 17:51:39 +0200 24834398-9335-475c-91d8-c4fac9c22cce
<![CDATA[Tibial bone and soft-tissue concentrations following combination therapy with vancomycin and meropenem – evaluated by microdialysis in a porcine model]]> Vittrup, S., Hanberg, P., Knudsen, M. B., et al. Aims Prompt and sufficient broad-spectrum empirical antibiotic treatment is key to preventing infection following open tibial fractures. Succeeding co-administration, we dynamically assessed the time for which vancomycin and meropenem concentrations were above relevant epidemiological cut-off (ECOFF) minimal inhibitory concentrations (T > MIC) in tibial compartments for the bacteria most frequently encountered in open fractures. Low and high MIC targets were applied: 1 and 4 µg/ml for vancomycin, and 0.125 and 2 µg/ml for meropenem. Methods Eight pigs received a single dose of 1,000 mg vancomycin and 1,000 mg meropenem simultaneously over 100 minutes and 10 minutes, respectively. Microdialysis catheters were placed for sampling over eight hours in tibial cancellous bone, cortical bone, and adjacent subcutaneous adipose tissue. Venous blood samples were collected as references. Results Across the targeted ECOFF values, vancomycin displayed longer T > MIC in all the investigated compartments in comparison to meropenem. For both drugs, cortical bone exhibited the shortest T > MIC. For the low MIC targets and across compartments, mean T > MIC ranged between 208 and 449 minutes (46% to 100%) for vancomycin and between 189 and 406 minutes (42% to 90%) for meropenem. For the high MIC targets, mean T > MIC ranged between 30 and 446 minutes (7% to 99%) for vancomycin and between 45 and 181 minutes (10% to 40%) for meropenem. Conclusion The differences in the T > MIC between the low and high targets illustrate how the interpretation of these results is highly susceptible to the defined MIC target. To encompass any trauma, contamination, or individual tissue differences, a more aggressive dosing approach may be considered to achieve longer T > MIC in all the exposed tissues, and thereby lower the risk of acquiring an infection after open tibial fractures.

Research Tue, 01 Feb 2022 17:51:39 +0100 a71c23f3-f849-400e-9f53-30e54007369c
<![CDATA[2D size of trabecular bone structure units (BSU) correlate more strongly with 3D architectural parameters than age in human vertebrae]]> Lamarche, B. A., Thomsen, J. S., Andreasen, C. M., Lievers, W. B., Andersen, T. L. Bone tissue is continuously remodeled. In trabecular bone, each remodeling transaction forms a microscopic bone structural unit (BSU), also known as a hemiosteon or a trabecular packet, which is bonded to existing tissue by osteopontin-rich cement lines. The size and shape of the BSUs are determined by the size and shape of the resorption cavity, and whether the cavity is potentially over- or under-filled by the subsequent bone formation. The present study focuses on the recently formed trabecular BSUs, and how their 2D size and shape changes with age and trabecular microstructure. The study was performed using osteopontin-immunostained frontal sections of L2 vertebrae from 8 young (aged 18.5–37.6 years) and 8 old (aged 69.1–96.4 years) control females, which underwent microcomputed tomography (μCT) imaging prior to sectioning. The contour of 4230 BSU profiles (181–385 per vertebra) within 1024 trabecular profiles were outlined, and their 2D width, length, area, and shape were assessed. Of these BSUs, 22 (0.5%) were generated by modeling-based bone formation (i.e. without prior resorption), while 99.5% were generated by remodeling-based bone formation (i.e. with prior resorption). The distributions of BSU profile width, length, and area were significantly smaller in the old versus young females (p < 0.005), and the median profile width, length, and area were negative correlated with age (p < 0.018). Importantly, these BSU profile size parameters were more strongly correlated with trabecular bone volume (BV/TV, p < 0.002) and structure model index (SMI, p < 0.008) assessed by μCT, than age. Moreover, the 2D BSU size parameters were positively correlated to the area of the individual trabecular profiles (p < 0.0001), which were significantly smaller in the old versus young females (p < 0.024). The BSU shape parameters (aspect ratio, circularity, and solidity) were not correlated with age, BV/TV, or SMI. Collectively, the study supports the notion that not only the BSU profile width, but also its length and area, are more influenced by the age-related bone loss and shift from plates to rods (SMI), than age itself. This implies that BSU profile size is mainly driven by changes in the trabecular microstructure, which affect the size of the resorption cavity that the BSU refills.

Research Fri, 01 Jul 2022 17:51:39 +0200 a11e8b00-6cb7-4465-8de5-415554402857
<![CDATA[Metabolite Profiling of the Social Spider <i>Stegodyphus dumicola </i>Along a Climate Gradient]]> Sandfeld, T. ., Malmos, K. G. ., Nielsen, C. B., et al. Research Tue, 01 Mar 2022 17:51:39 +0100 edd66e6a-a21f-40ac-b013-93cba6f1fa33 <![CDATA[The incidence of psychoactive substances and alcohol among impaired drivers in Denmark in 2015–2019.]]> Simonsen, K. W., Hasselstrøm, J. B., Hermansen, S. K., et al. This study examines the presence of psychoactive drugs and alcohol in blood from apprehended drivers driving under the influence of drugs (DUID) and alcohol in Denmark in a five-year period from 2015 to 2019. Data were analysed with respect to gender, age, substances with concentrations above the Danish legal limit, arresting time of day and repeat arrest. By request of the police, the blood samples were subjected to analysis for alcohol and/or tetrahydrocannabinol (THC) alone, for “other drugs” (covering all drugs including new psychoactive substances (NPS), except THC, listed in the Danish list of narcotic drugs) or for both THC and other drugs. About the same number of alcohol traffic cases (37,960) and drug traffic cases (37,818) were submitted for analysis for the five-year period. The number of drug traffic cases per year increased from 5660 cases in 2015 to 9505 cases in 2019, while the number of alcohol traffic cases per year (average, 7600) was unchanged. Ethanol (89.2%) was the overall most frequent single substance, followed by THC (68.2%). CNS stimulants (46.8%) were the second most prevalent group of non-alcoholic drugs. Cocaine (23.8%) and amphetamine (22.9%) were the most frequent CNS stimulants. The proportion of CNS-stimulant positive drivers more than doubled in ten years. Benzodiazepines/z-hypnotics (12.7%) were the third most prevalent drug group detected, with clonazepam (8%) as the most frequent drug. Opioids were above the legal limit in 9.8% of the cases. NPS was above the legal limit in 128 cases (0.6%). Poly-drug use occurred in 40% of the DUID cases in the requested groups: other drug or other drug/THC. Young males dominated the DUID cases (median age 26). Drink-drivers (median age 39) were also mainly men, but the age distribution was equally spread over the age groups. Re-arrest occurred more often in DUID drivers (18–29%) than in drinking drivers (6–12%). DUID was evenly spread over the week, while drink-driving was most frequent on weekends. This study is an important supplement to the knowledge of drug use in Denmark. It was the well-known psychoactive substances that were detected. Only a few NPS occurred. However, the abuse pattern has changed, and CNS stimulants now account for a much higher proportion than earlier. Our results indicate a drug use problem among DUID drivers. This gives rise to concern because of a risk of traffic accidents. Treating the underlying abuse problem is therefore recommended, rather than focusing solely on prosecuting.

Research Fri, 01 Apr 2022 17:51:39 +0200 f84bfb83-c5c0-4b86-bf8b-f7fe00095d1d
<![CDATA[Hyperbaric Oxygen Treatment for Diabetic Retinopathy and Neuropathy in a Streptozotocin Induced Diabetic Rat Model]]> Madsen, J. G., Skov, M. N., Hansen, K., et al. Research Wed, 01 Dec 2021 17:51:39 +0100 90ebacfd-ba1b-4f54-a8b6-3b60baf0f5d8 <![CDATA[A detachable tourniquet to aid rodent tail-vasculature catheterisation; especially useful for preclinical imaging]]> Hansen, K. Research Fri, 03 Sep 2021 17:51:39 +0200 d835b3ee-a10f-4d82-8cb7-5b8f5519b63b <![CDATA[Quantitative analysis of pulmonary structures in PMCT; Stereological comparison of drowning compared to opioid-overdose cases]]> Jakobsen, S. R., Borg Hansen, I., Harders, S. W., et al. The aim of this study, was to introduce stereology as a versatile and robust tool for quantitative image analysis of volume and attenuation characteristics (Hounsfield Units (HU's)), in a blinded control case study investigating lungs of drowning victims compared to a control group on post mortem computed tomography (PMCT) data.

Materials and Methods
PMCT scans of the lungs from 14 drowned cases and 14 matched opioid-overdose-controls was included. Quantitative CT-analysis was performed using a stereological approach adapted to PMCT data that allowed for precise extraction of volume and HU-values using stereological point-probes assigned manually to individual lung-structures. Qualitative radiological image interpretation performed by a trained radiologist was compared to the quantitative analyses.

No significant difference was found for total lung volume, volumes of consolidations, ground glass opacities, bronchi, and air-filled lung tissue. When comparing drowning cases with opioid overdose cases as controls, the extracted HU-values did not show statistically significant changes in mean attenuation characteristics. No major discrepancies were found between the quantitative analysis and qualitative analysis.

Conventional radiological evaluation of PMCT images rely on the radiologists’ ability to distinguish normal from pathological. Quantitative image analysis offers, to name a few, precise estimations of structure volume and HU-statistics. Although the used 14 matched cases data failed to significantly aid the diagnosis of drowning statistically, we envisage that quantitative PMCT analysis using stereology could become a valuable tool to improve objective forensic radiological interpretation.]]>
Research Tue, 01 Mar 2022 17:51:39 +0100 0e3c8529-f74c-424f-95d1-b5a282ff8e3a
<![CDATA[NOMI]]> Rædkjær, M. Research Fri, 05 Nov 2021 17:51:39 +0100 536fadd8-5e39-4575-91b5-3e366c7c1df2 <![CDATA[Asphyxia homicides in Denmark 1992-2016]]> Thomsen, A. H., Leth, P. M., Hougen, H. P., Villesen, P. In this retrospective study , we present the findings in 250 homicides by asphyxia in Denmark in a 25-year period, with a particular focus on the autopsy findings in strangulation. Our intention is for the results to be used in future death investigations, where difficulties in interpretation of findings in potential asphyxial deaths arise. Asphyxia homicides showed a strong bias with respect to sex, age, and homicide type. The frequent female victim was typically an adult, whereas the rarer male victim was most often a child. Female offenders most often killed their children, and male offenders most often killed their female partner. Generally, most asphyxia homicides took place in a domestic setting. Manual strangulation and ligature strangulation were the most common mechanisms of asphyxia homicides (81.6%). A lack of petechial hemorrhages, especially in the conjunctiva, was rare in homicidal strangulation, but there were exceptions, especially when there was postmortem decomposition, making it impossible to verify them. Most victims of strangulation had skin lesions in the face (including the jawline) or on the neck, with accompanying hemorrhages in muscle and connective tissue, but the findings could be subtle or compounded by decomposition. Fractures of the laryngo-hyoid complex were common in strangulation, particularly in manual strangulation (chi-sq = 4.0993, df = 1, P < 0.05) and were clearly related to the age of the victim (chi-sq = 82.193, df = 4, P < 0.001). In children and young adults dying from homicidal strangulation, a lack of fractures is to be expected, while a lack of fractures is unusual, but not entirely unexpected, for adults and aged people.

Research Tue, 01 Nov 2022 17:51:39 +0100 e4e09741-9f66-4afa-a971-022841a968f5
<![CDATA[The effect of casein glycomacropeptide versus free synthetic amino acids for early treatment of phenylketonuria in a mice model]]> Ahring, K. K., Dagnæs-Hansen, F., Brüel, A., et al. Introduction Management of phenylketonuria (PKU) is mainly achieved through dietary control with limited intake of phenylalanine (Phe) from food, supplemented with low protein (LP) food and a mixture of free synthetic (FS) amino acids (AA) (FSAA). Casein glycomacropeptide (CGMP) is a natural peptide released in whey during cheese making by the action of the enzyme chymosin. Because CGMP in its pure form does not contain Phe, it is nutritionally suitable as a supplement in the diet for PKU when enriched with specific AAs. Lacprodan® CGMP-20 (= CGMP) used in this study contained only trace amounts of Phe due to minor presence of other proteins/peptides. Objective The aims were to address the following questions in a classical PKU mouse model: Study 1, off diet: Can pure CGMP or CGMP supplemented with Large Neutral Amino Acids (LNAA) as a supplement to normal diet significantly lower the content of Phe in the brain compared to a control group on normal diet, and does supplementation of selected LNAA results in significant lower brain Phe level?. Study 2, on diet: Does a combination of CGMP, essential (non-Phe) EAAs and LP diet, provide similar plasma and brain Phe levels, growth and behavioral skills as a formula which alone consist of FSAA, with a similar composition?. Material and methods 45 female mice homozygous for the Pahenu2 mutation were treated for 12 weeks in five different groups; G1(N-CGMP), fed on Normal (N) casein diet (75%) in combination with CGMP (25%); G2 (N-CGMP-LNAA), fed on Normal (N) casein diet (75%) in combination with CGMP (19,7%) and selected LNAA (5,3% Leu, Tyr and Trp); G3 (N), fed on normal casein diet (100%); G4 (CGMP-EAA-LP), fed on CGMP (70,4%) in combination with essential AA (19,6%) and LP diet; G5 (FSAA-LP), fed on FSAA (100%) and LP diet. The following parameters were measured during the treatment period: Plasma AA profiles including Phe and Tyr, growth, food and water intake and number of teeth cut. At the end of the treatment period, a body scan (fat and lean body mass) and a behavioral test (Barnes Maze) were performed. Finally, the brains were examined for content of Phe, Tyr, Trp, dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC), serotonin (5-HT) and 5-hydroxyindole-acetic acid (5-HIAA), and the bone density and bone mineral content were determined by dual-energy x-ray absorptiometry. Results Study 1: Mice off diet supplemented with CGMP (G1 (N-CGMP)) or supplemented with CGMP in combination with LNAA (G2 (N-CGMP-LNAA)) had significantly lower Phe in plasma and in the brain compared to mice fed only casein (G3 (N)). Extra LNAA (Tyr, Trp and Leu) to CGMP did not have any significant impact on Phe levels in the plasma and brain, but an increase in serotonin was measured in the brain of G2 mice compared to G1. Study 2: PKU mice fed with mixture of CGMP and EAA as supplement to LP diet (G4 (CGMP-EAA-LP)) demonstrated lower plasma-Phe levels but similar brain- Phe levels and growth as mice fed on an almost identical combination of FSAA (G5 (FSAA-LP)). Conclusion CGMP can be a relevant supplement for the treatment of PKU.

Research Sat, 01 Jan 2022 17:51:39 +0100 e3729999-5996-45fb-b7e6-58ac73919dab
<![CDATA[Oral lactate slows gastric emptying and suppresses appetite in young males]]> Pedersen, M. G. B., Søndergaard, E., Nielsen, C. B., et al. BACKGROUND: Lactate serves as an alternative energy fuel but is also an important signaling metabolite. We aimed to investigate whether oral lactate administration affects appetite-regulating hormones, slows gastric emptying rate, and dampens appetite.

METHODS: Ten healthy male volunteers were investigated on two separate occasions: 1) following oral ingestion of D/L-Na-lactate and 2) following oral ingestion of isotonic iso-voluminous NaCl and intravenous iso-lactemic D/L-Na-lactate infusions. Appetite was evaluated by questionnaires and ad libitum meal tests were performed at the end of each study day. Gastric emptying rate was evaluated using the acetaminophen test.

RESULTS: Plasma concentrations of growth differential factor 15 (GDF15, primary outcome) increased following oral and iv administration of lactate (p < 0.001) with no detectable difference between interventions (p = 0.15). Oral lactate administration lowered plasma concentrations of acylated ghrelin (p = 0.02) and elevated glucagon like peptide-1 (GLP-1, p = 0.045), insulin (p < 0.001), and glucagon (p < 0.001) compared with iv administration. Oral lactate administration slowed gastric emptying (p < 0.001), increased the feeling of being "full" (p = 0.008) and lowered the "anticipated future food intake" (p = 0.007) compared with iv administration. Food intake during the ad libitum meal test did not differ between the two study days.

CONCLUSION: Oral lactate administration has a direct effect on the upper gastrointestinal tract, affecting gut hormone secretion, motility and appetite sensations which cannot be mediated through lactate in the systemic circulation alone. These data suggest that compounds rich in lactate may be useful in the treatment of metabolic disease.


Research Tue, 01 Feb 2022 17:51:39 +0100 d0a29638-d7ac-4dec-9b2c-81401b0afb85
<![CDATA[The Mechanism Switching the Osteoclast From Short to Long Duration Bone Resorption]]> Delaisse, J. M., Søe, K., Andersen, T. L., Rojek, A. M., Marcussen, N. The current models of osteoclastic bone resorption focus on immobile osteoclasts sitting on the bone surface and drilling a pit into the bone matrix. It recently appeared that many osteoclasts also enlarge their pit by moving across the bone surface while resorbing. Drilling a pit thus represents only the start of a resorption event of much larger amplitude. This prolonged resorption activity significantly contributes to pathological bone destruction, but the mechanism whereby the osteoclast engages in this process does not have an answer within the standard bone resorption models. Herein, we review observations that lead to envision how prolonged resorption is possible through simultaneous resorption and migration. According to the standard pit model, the “sealing zone” which surrounds the ruffled border (i.e., the actual resorption apparatus), “anchors” the ruffled border against the bone surface to be resorbed. Herein, we highlight that continuation of resorption demands that the sealing zone “glides” inside the cavity. Thereby, the sealing zone emerges as the structure responsible for orienting and displacing the ruffled border, e.g., directing resorption against the cavity wall. Importantly, sealing zone displacement stringently requires thorough collagen removal from the cavity wall - which renders strong cathepsin K collagenolysis indispensable for engagement of osteoclasts in cavity-enlargement. Furthermore, the sealing zone is associated with generation of new ruffled border at the leading edge, thereby allowing the ruffled border to move ahead. The sealing zone and ruffled border displacements are coordinated with the migration of the cell body, shown to be under control of lamellipodia at the leading edge and of the release of resorption products at the rear. We propose that bone resorption demands more attention to osteoclastic models integrating resorption and migration activities into just one cell phenotype.

Research Mon, 01 Mar 2021 17:51:39 +0100 34ff9ebe-ffb2-441d-bf74-2d30c84dd398
<![CDATA[A novel nonsense variant in MED12 associated with malformations in a female fetus]]> Faergeman, S. L., Becher, N., Andreasen, L., Christiansen, M., Frost, L., Vogel, I. Pathogenic variants in the MED12 gene located on the X-chromosome have primarily been reported in males with Lujan-Fryns syndrome, Ohdo syndrome and the Opits-Kaveggia syndrome. However, earlier reports of female patients and female mice suggest that MED12 deficiency causes severe malformations. We report a novel example of a MED12 de novo nonsense variant in a female fetus with severe malformations identified by trio-exome sequencing. This finding further expands the clinical spectrum of MED12-related disorders, which is vital for prenatal diagnosis and genetic counselling of couples.

Research Wed, 01 Dec 2021 17:51:40 +0100 885bf84d-7373-4981-b754-2a074521fb80
<![CDATA[Advanced magnetic resonance imaging of chronic whiplash patients]]> Uhrenholt, L., Brix, L., Wichmann, T. O., Pedersen, M., Ringgaard, S., Jensen, T. S. BACKGROUND: Whiplash injury is common following road traffic crashes affecting millions worldwide, with up to 50% of the injured developing chronic symptoms and 15% having a reduced working capability due to ongoing disability. Many of these patients receive treatment in primary care settings based upon clinical and diagnostic imaging findings. Despite the identification of different types of injuries in the whiplash patients, clinically significant relationships between injuries and chronic symptoms remains to be fully established. This study investigated the feasibility of magnetic resonance imaging (MRI) techniques including quantitative diffusion weighted imaging and measurements of cerebrospinal fluid (CSF) flow as novel non-invasive biomarkers in a population of healthy volunteers and chronic whiplash patients recruited from a chiropractic clinic for the purpose of improving our understanding of whiplash injury.

METHODS: Twenty chronic whiplash patients and 18 healthy age- and gender matched control subjects were included [mean age ± SD (sex ratio; females/males), case group: 37.8 years ± 9.1 (1.22), control group: 35.1 years ± 9.2 (1.25)]. Data was collected from May 2019 to July 2020. Data from questionnaires pertaining to the car crash, acute and current symptoms were retrieved and findings from clinical examination and MRI including morphologic, diffusion weighted and phase-contrast images were recorded. The apparent diffusion coefficient and fractional anisotropy were calculated, and measurement and analysis of CSF flow was conducted. Statistical analyses included Fisher's exact test, Mann Whitney U test and analysis of variance between groups.

RESULTS: The studied population was described in detail using readily available clinical tools. No statistically significant differences were found between the groups on MRI.

CONCLUSIONS: This study did not show that MRI-based measures of morphology, spinal cord and nerve root diffusion or cerebrospinal fluid flow are sensitive biomarkers to distinguish between chronic whiplash patients and healthy controls. The detailed description of the chronic whiplash patients using readily available clinical tools may be of great relevance to the clinician. In the context of feasibility, clinical practice-based advanced imaging studies with a technical setup similar to the presented can be expected to have a high likelihood of successful completion.

Research Sat, 01 Jan 2022 17:51:40 +0100 0903e236-3527-4f96-a352-475bea0d7076
<![CDATA[Human hematopoietic microenvironments]]> Kristensen, H. B., Andersen, T. L., Patriarca, A., et al. Dormancy of hematopoietic stem cells and formation of progenitors are directed by signals that come from the bone marrow microenvironment. Considerable knowledge has been gained on the murine hematopoietic stem cell microenvironment, while less so on the murine progenitor microenvironment and even less so on these microenvironments in humans. Characterization of these microenvironments is decisive for understanding hematopoiesis and finding new treatment modalities against bone marrow malignancies in the clinic. However, it is equally challenging, because hematopoietic stem cells are difficult to detect in the complex bone marrow landscape. In the present study we are characterizing the human hematopoietic stem cell and progenitor microenvironment. We obtained three adjacent bone marrow sections from ten healthy volunteers. One was used to identify a population of CD34+/CD38- “hematopoietic stem cells and multipotent progenitors” and a population of CD34+/CD38+ “progenitors” based on immunofluorescence pattern/intensity and cellular morphology. The other two were immunostained respectively for CD34/CD56 and for CD34/SMA. Using the combined information we performed a non-computer-assisted quantification of nine bone marrow components (adipocytes, megakaryocytes, bone surfaces, four different vessel types (arteries, capillaries, sinusoids and collecting sinuses), other “hematopoietic stem cells and multipotent progenitors” and other “progenitors”) within 30 μm of “hematopoietic stem cells and multipotent progenitors”, “progenitors”, and “random cell profiles”. We show that the microenvironment of the “hematopoietic stem cells and multipotent progenitors” is significantly enriched in sinusoids and megakaryocytes, while the microenvironment of the “progenitors” is significantly enriched in capillaries, other “progenitors”, bone surfaces and arteries.

Research Thu, 01 Apr 2021 17:51:40 +0200 7ad3d02d-3939-472f-9a71-6ed484feceb2
<![CDATA[Trafik]]> Uhrenholt, L. Education Fri, 01 Jan 2021 17:51:40 +0100 02783e78-df6e-403d-9ae0-04e2a4aadcf3 <![CDATA[Lethal abusive head trauma in infancy in Denmark from 2000 to 2011]]> Flugt, A., Frost, L., Søndergaard, C., Milidou, I. Research Mon, 01 Mar 2021 17:51:40 +0100 02616127-bb87-423d-9589-e011bfe6af64 <![CDATA[Purity of street-level cocaine across Denmark from 2006 to 2019]]> Hesse, M., Thomsen, K. R., Thylstrup, B., et al. Cocaine-related emergency department admissions are increasing, and cocaine seizures are at an all-time high in Europe. Our aim was to investigate the trends in purity and adulterants over time in cocaine available to cocaine users at street level in Denmark. We used a representative sample of cocaine seized at street level and analyzed by the national departments of forensic medicine between 2006 and 2019 (n = 1460). Latent profile analysis was used to classify the samples based on cocaine, levamisole, and phenacetin content. Low purity cocaine comprised most of the cocaine seizures in early years, but its share began to decline in 2013, and from 2016 to 2019, the high purity profile was dominant. While the total number of samples containing adulterants decreased, levamisole remained a common and dangerous adulterant. The findings underline the need to inform the public, medical doctors, and service providers for people with drug use disorders about the higher potency of street cocaine.

Research Wed, 01 Dec 2021 17:51:40 +0100 df40cb61-ece4-4b35-9bbb-fafd3cce3557
<![CDATA[Assessment of XCMS Optimization Methods with Machine-Learning Performance]]> Lassen, J., Nielsen, K. L., Johannsen, M., Villesen, P. The metabolomics field is under rapid development. In particular, biomarker identification and pathway analysis are growing, as untargeted metabolomics is usable for discovery research. Frequently, new processing and statistical strategies are proposed to accommodate the increasing demand for robust and standardized data. One such algorithm is XCMS, which processes raw data into integrated peaks. Multiple studies have tried to assess the effect of optimizing XCMS parameters, but it is challenging to quantify the quality of the XCMS output. In this study, we investigate the effect of two automated optimization tools (Autotuner and isotopologue parameter optimization (IPO)) using the prediction power of machine learning as a proxy for the quality of the data set. We show that optimized parameters outperform default XCMS settings and that manually chosen parameters by liquid chromatography-mass spectrometry (LC-MS) experts remain the best. Finally, the machine-learning approach of quality assessment is proposed for future evaluations of newly developed optimization methods because its performance directly measures the retained signal upon preprocessing.

Research Fri, 01 Oct 2021 17:51:40 +0200 20da8d66-2f91-40d0-9374-9df153f824ce
<![CDATA[Guidelines for Biobanking of Bone Marrow Adipose Tissue and Related Cell Types]]> Lucas, S., Tencerova, M., von der Weid, B., et al. Over the last two decades, increased interest of scientists to study bone marrow adiposity (BMA) in relation to bone and adipose tissue physiology has expanded the number of publications using different sources of bone marrow adipose tissue (BMAT). However, each source of BMAT has its limitations in the number of downstream analyses for which it can be used. Based on this increased scientific demand, the International Bone Marrow Adiposity Society (BMAS) established a Biobanking Working Group to identify the challenges of biobanking for human BMA-related samples and to develop guidelines to advance establishment of biobanks for BMA research. BMA is a young, growing field with increased interest among many diverse scientific communities. These bring new perspectives and important biological questions on how to improve and build an international community with biobank databases that can be used and shared all over the world. However, to create internationally accessible biobanks, several practical and legislative issues must be addressed to create a general ethical protocol used in all institutes, to allow for exchange of biological material internationally. In this position paper, the BMAS Biobanking Working Group describes similarities and differences of patient information (PIF) and consent forms from different institutes and addresses a possibility to create uniform documents for BMA biobanking purposes. Further, based on discussion among Working Group members, we report an overview of the current isolation protocols for human bone marrow adipocytes (BMAds) and bone marrow stromal cells (BMSCs, formerly mesenchymal), highlighting the specific points crucial for effective isolation. Although we remain far from a unified BMAd isolation protocol and PIF, we have summarized all of these important aspects, which are needed to build a BMA biobank. In conclusion, we believe that harmonizing isolation protocols and PIF globally will help to build international collaborations and improve the quality and interpretation of BMA research outcomes.

Research Wed, 01 Sep 2021 17:51:40 +0200 f6a1ef29-fd42-4d62-a07e-2f0714654eb6
<![CDATA[Myeloma-bone marrow adipocyte axis in tumour survival and treatment response]]> Jafari, A., Fairfield, H., Andersen, T. L., Reagan, M. R. Multiple myeloma is an incurable cancer of the bone marrow that is dependent on its microenvironment, including bone marrow adipocytes (BMAds). Here, we discuss our findings that the reciprocal interaction of myeloma cells and BMAds, leads to myeloma cell survival and induces metabolic dysfunction and senescence-associated secretory phenotype in BMAds.

Research Wed, 01 Sep 2021 17:51:40 +0200 d57f615a-8196-422a-9b4a-12f9be31b174
<![CDATA[Stability investigations of cytochrome P450 (CYP) enzymes immediately after death in a pig model support the applicability of postmortem hepatic CYP quantification]]> Pedersen, K. W., Hansen, J., Hasselstrøm, J. B., Jornil, J. R. Quantification of drug-metabolizing cytochrome P450 (CYP) isoforms using LC-MS/MS has been proposed as a potential way of estimating antemortem CYP levels using postmortem tissue, but the postmortem stability of CYP proteins is incompletely investigated. If one can use data obtained from the analysis of postmortem specimens to inform physiologically based pharmacokinetic (PBPK) models this greatly increases the access to rare specimens among special subpopulations. In this study, we developed and validated an LC-MS/MS method for targeted CYP protein quantification in a porcine animal model to study postmortem stability. We measured 19.9-28.3 pmol CYP1A2, 50.3-66.2 pmol CYP2D25, 132.9-142.7 pmol CYP2E1, and 16.8-48 pmol CYP3A29 protein per mg PLM in nondegraded tissue. In tissue stored at 4°C, we found that the CYP protein levels were unaffected by degradation after 72 h. At 21°C CYP1A2, CYP2D25, and CYP2E1 protein levels were nearly unaffected by degradation after 24 h, whereas a loss of approximately 50% was seen after 48 h. At 21°C CYP3A29 had a loss of 50% at 24 h and 70% at 48 h exhibiting less postmortem stability. In vitro enzyme activity measurements in the same tissue stored at 21°C showed a 50% decrease after 24 h and a complete loss of enzyme activity after 48 h. When stored at 4°C, the in vitro enzyme activity decreased to 50% activity after 96 h. In conclusion, measuring CYP levels by an LC-MS/MS approach was clearly less affected by postmortem changes than an activity-based approach. The found postmortem stability for 24 h at 21°C for 3 out of 4 CYP isoforms supports the use of properly stored postmortem tissue to inform PBPK models.

Research Fri, 01 Oct 2021 17:51:40 +0200 22643fa9-318e-40e4-946a-edb9ac17edca
<![CDATA[Immune checkpoint inhibitor-induced myocarditis in cancer patients]]> Matzen, E., Bartels, L. E., Løgstrup, B., Horskær, S., Stilling, C., Donskov, F. Background: Immune checkpoint inhibitor (ICI) induced myocarditis is a rare, severe, and often fatal adverse event. Evidence to guide appropriate immunosuppressive therapy is scarce. We present a case of ICI-induced myocarditis and a review of ICI-induced myocarditis cases to determine the most effective immunosuppressive therapeutic strategy for ICI-induced myocarditis. Methods: A systematic search of PubMed was carried out for treatment of ICI-induced myocarditis. Reference lists from identified articles were manually reviewed for additional cases. Results: A total of 87 cases with ICI-induced myocarditis were identified. The majority were melanoma (n = 39), lung cancer (n = 19), renal cell cancer (n = 10), and thymoma cancer patients (n = 4). In 38 (44%) cases, patients received high-dose steroid treatment only. A total of 49 (56%) cases were treated with immunosuppressive agents other than steroid; a total of 13 different immunosuppressive agents were used, including alemtuzumab or abatacept. The median time to onset of symptoms after initiation of ICI was 16 days (range, 1–196 days); cardiotoxic symptoms developed after 2 cycles of ICI (range, 1–13 cycles). A total of 48% of cases were fatal. In cases treated with high-dose steroids only vs. cases treated with other immunosuppressive agents, fatality was 55% and 43% respectively. In 64 out of the 87 cases, tumor control was not described. In patients treated with high-dose steroids only, two patients had stable disease as best tumor response; in patients treated with other immunosuppressive agents, one complete response, one partial response and seven stable disease were noted as best tumor response. Overall, 11 studies were at low risk of bias (12.6%), 38 at moderate risk of bias (43.7%) and 38 at high risk of bias (43.7%). Conclusion: Immune checkpoint inhibitor induced myocarditis is a serious and often fatal adverse event. High-dose prednisolone, alemtuzumab or abatacept are all possible treatments options for ICI-induced myocarditis, whereas infliximab increases the risk of death from cardiovascular causes, and should be avoided. Further research is needed.

Research Wed, 01 Dec 2021 17:51:40 +0100 3078a315-a0c5-4816-8ee9-a5862291e8a8
<![CDATA[Dödligt skjutvapenvåld i Sverige och andra europeiska länder]]> Aplowski, A., Askeland, O. M. ., Birkel, C. ., et al. Research Fri, 01 Jan 2021 17:51:40 +0100 cee713c2-6372-46ac-ac34-4a0affef5144 <![CDATA[Human calculus – et omfattende kartotek af informationer om livsstil og arvemasse]]> Staun Larsen, L., Bindslev, D. A. Communication Fri, 01 Jan 2021 17:51:40 +0100 b30705f4-1a41-4579-9b14-6a7b4c42b8cd <![CDATA[The PERFORM-P (Principles of Evidence-based Reporting in FORensic Medicine-Pathology version)]]> Meilia, P. D.I., Herkutanto, ., Atmadja, D. S., et al. Introduction: Most findings of forensic pathology examinations are presented as written reports. There are currently no internationally accepted recommendations for writing forensic pathology reports. Existing recommendations are also varied and reflect the differences in the scope and role of forensic medical services and local settings in which they are to be implemented. The legal fact-finder thus faces wide variation in the quality of forensic pathology reports, which poses a threat to the reliability of legal decision-making. To address this issue, the development of the “PERFORM-P (Principles of Evidence-based Reporting in FORensic Medicine-Pathology version)” was undertaken. The goal of the PERFORM-P is to provide common practice recommendations adaptable to local requirements to promote evidence-based practice (EBP) in forensic pathology. Methods: An international consensus study was conducted in three phases by (1) developing a long-list of items to be considered in the reporting recommendations, (2) conducting a Delphi process (an iterative survey method to transform individual opinions into group consensus) with international forensic pathologists, and (3) designing the PERFORM-P prototype and its accompanying manual. Results: With assistance from 106 forensic pathologists/forensic medical practitioners from 41 countries, the PERFORM-P was developed. The PERFORM-P consists of a list of 61 items to be included in a forensic pathology report, which is accompanied by its Explanation and Elaboration (E&E) document. Discussion: To prepare forensic pathology (postmortem) reports that incorporate principles of evidence-based practice, internationally accepted recommendations might be helpful. The PERFORM-P identifies recommendations for necessary elements to include in a forensic pathology report. PERFORM-P can be applied to a wide range of matters requiring forensic pathological analysis, acceptable to forensic pathologists from a representative selection of jurisdictions and medico-legal systems.

Research Fri, 01 Oct 2021 17:51:40 +0200 b02a9517-c352-433a-a5bf-01f58057b7a9
<![CDATA[Looking deep into the past – virtual autopsy of a Mongolian warrior]]> Pedersen, C. C. E., Villa, C., Asingh, P., Thali, M. J., Gascho, D. Introduction: In 2016, a well-preserved mummy was discovered in the cold and dry climate of the Altai Mountains in the western part of Mongolia. The mummy was thought to have been a mounted warrior based on its riding boots and the saddle, bow and arrow found beside the body. A virtual autopsy was approved to obtain additional information on the body's internal structure and to possibly determine the cause of death. Materials and Methods: A computed tomography (CT) scan was performed on the Mongolian mummy using a 64-slice CT scanner. In addition to multiplanar reconstructions, volume and cinematic rendering techniques were applied for three-dimensional visualization. Results: The shape of the pelvis indicated male sex. The skull demonstrated a flat facial profile, common among ancient Mongolian populations. Signs of periodontitis were detected in one of the six remaining teeth. The intact hyoid and thyroid were visible on the CT images together with the cartilaginous parts of the trachea, which excluded a laryngeal fracture. Signs of degenerative alterations (Schmorl´s nodes and osteophytes) were identified in the lumbar spine, attributed to regular which might be a sign of skeletal changes induced by horse riding. Indications for a violent cause of death were not detected. Conclusion: Through CT examination, some degenerative changes and pathological findings could be detected noninvasively. Although no cause of death could be determined, radiodiagnostic findings, such as degenerative alterations in the lumbar spine, were relevant, for instance, supporting the original hypothesis of a mounted warrior based on the saddle and riding boots found.

Research Tue, 01 Jun 2021 17:51:40 +0200 cb1c5469-b522-4365-bea0-0fe79102897c
<![CDATA[Genetic investigations of 100 inherited cardiac disease-related genes in deceased individuals with schizophrenia]]> Christiansen, S. L., Andersen, J. D., Themudo, G. E., et al. Cardiac diseases and sudden cardiac death (SCD) are more prevalent in individuals diagnosed with schizophrenia compared to the general population, with especially coronary artery disease (CAD) as the major cardiovascular cause of death. Antipsychotic medications, genetics, and lifestyle factors may contribute to the increased SCD in individuals with schizophrenia. The role of antipsychotic medications and lifestyle factors have been widely investigated, while the genetic predisposition to inherited cardiac diseases in schizophrenia is poorly understood. In this study, we examined 100 genes associated with inherited cardiomyopathies and cardiac channelopathies in 97 deceased individuals diagnosed with schizophrenia for the prevalence of genetic variants associated with SCD. The deceased individuals had various causes of death and were included in the SURVIVE project, a prospective, autopsy-based study of mentally ill individuals in Denmark. This is the first study of multiple inherited cardiac disease-related genes in deceased individuals with diagnosed schizophrenia to shed light on the genetic predisposition to SCD in individuals with schizophrenia. We found no evidence for an overrepresentation of rare variants with high penetrance in inherited cardiac diseases, following the American College of Medical Genetics and Genomics and the Association for Molecular Pathology (ACMG) consensus guidelines. However, we found that the deceased individuals had a statistically significantly increased polygenic burden caused by variants in the investigated heart genes compared to the general population. This indicates that common variants with smaller effects in heart genes may play a role in schizophrenia.

Research Thu, 01 Jul 2021 17:51:40 +0200 5adbd82a-3a3e-4c13-b6c2-86a782b55759
<![CDATA[Active Blood Acidification Greatly Enhances Oxygen Supply to the Teleost Retina]]> Damsgaard, C., Lauridsen, H., Harter, T., et al. Research Sat, 01 May 2021 17:51:40 +0200 e66fd5cf-6f08-446e-a8c4-a55c19a74601 <![CDATA[The influence of assisted ventilation and recumbency on cardiorespiratory physiology in the anesthetized freshwater turtle Trachemys scripta scripta]]> Williams, C. J.A., Hansen, K., Williams, N., et al. The use of assisted ventilation is required in anesthetized reptiles as their respiratory drive is lost at surgical depths of anesthesia. The minute volume of the assisted ventilation influences arterial blood gases and acid-base regulation. Meanwhile, the ventilatory pattern may also affect hemodynamics in chelonians, which, given their large capacity for cardiac shunts, may impact the efficacy of the ventilation in terms of gas exchange. Hence, there is a need for primary information on the influence of assisted ventilation on chelonian physiology, and we, therefore, performed a randomized study into the effects of recumbency and maximum airway pressure on pressure-cycled ventilation in nine female Trachemys scripta scripta. Pronounced effects of ventilation pressure on arterial PCO2 and pH regardless of recumbency were revealed, whilst dorsal recumbency led to a larger Arterial-alveolar (A-a) O2 difference, suggesting compromised pulmonary gas exchange. Plasma [Na+] and [K+] balance was also significantly correlated with maximum airway pressure. Computed tomography (CT) scanning at a range of end-inspiratory pressures and ventral and dorsal recumbencies in eight T. scripta scripta showed that lung volumes increase with maximum ventilatory pressure, while recumbency did not influence volume at pressures above 5 cmH2O. Static compliance of the lungs was influenced by recumbency at neutral pressures. In conclusion, dorsal recumbency reduces pulmonary efficacy during positive pressure ventilation and tends to lower lung volume when ventilation is not provided. However, lung volumes and function - even in dorsal recumbency - can be adequately supported by assisted ventilation, and an end inspiratory pressure of 10 cmH2O at 4 breaths min−1 provided the most physiologically appropriate ventilation of anesthetized T. scripta scripta.

Research Fri, 01 Oct 2021 17:51:40 +0200 bf3e8037-4a53-407f-ba6a-9cf7aa47134c
<![CDATA[Identifying signs of child neglect and abuse in general practice]]> Merrild, C. H., Frost, L. INTRODUCTION: Children who live with neglect and abuse are often identified late in the process. At the front line of Danish healthcare, where most children are seen regularly, general practice is well placed to raise concerns about child health and wellbeing. Little is known about the role general practitioners (GPs) play in suspecting and reporting child neglect and abuse. We explored challenges GPs are facing in identifying such children and illustrated some of the barriers preventing GPs from reporting on these cases. METHODS: This was an explorative pilot study, preceding a larger multidisciplinary project. We conducted eight semi-structured interviews with selected Danish GPs. The interviews were transcribed verbatim and coded using thematic analysis. RESULTS: GPs rarely experienced concrete signs of child neglect and abuse, and reporting to the social services was often a way of helping families to get the support they needed. When GPs suspected that “something was wrong”, this was based on a gut feeling, triggered by non-measurable and intangible signs such as changes in health-seeking behaviour or in the relationship between caregivers and children. CONCLUSIONS: The intangibility of signs provoking suspicion of neglect and abuse made acting or reporting difficult and GPs felt that they lacked opportunities to take action. More knowledge is needed on how to approach matters of child protection and wellbeing across health professions and specialities.

Research Fri, 01 Jan 2021 17:51:40 +0100 4e4dad8f-83dd-4776-89ce-4c5577cac8dd
<![CDATA[Current advances in CT imaging of the deceased lung]]> Hansen, K., Morgan, B. Lung dysfunction causes significant mortality and morbidity and is a key organ in post-mortem investigation. Post-mortem (PM) medical imaging is being increasingly used, with computed tomography (CT) the most widespread modality. PMCT is used either as a supplement or an alternative to autopsy, especially in cases where autopsy is not possible or undesirable. Unfortunately, natural PM-processes in the lung obscure radiological interpretation in the dead, which accordingly is very different from the living. Interpretation of PMCT therefore requires special training and experience. The lung is also relatively difficult to examine during conventional autopsy, but diagnosis is improved by histological processing to allow ex vivo identification of infections and other pathological conditions. PMCT can examine the lung in-situ and is even superior to autopsy in certain aspects (especially for pneumothorax, small nodules and showing extent of abnormalities). Ventilated PMCT can be used to mimic clinical breath-hold procedures to improve overall pulmonary imaging quality.

Research Sun, 01 Aug 2021 17:51:40 +0200 a9914ce3-1805-445d-8b97-b458fdf52cc5
<![CDATA[The role of QT-prolonging medications in a forensic autopsy study from Western Denmark]]> Ahmed, H., Larsen, M. K., Hansen, M. R., Andersen, C. U. Medication-induced prolongation of the QT-interval (miQTP) can lead to cardiac arrhythmia. Our aim was to investigate the prevalence of forensic autopsy cases where fatal cardiac arrhythmia related to treatment with QT-prolonging medications (QT-PMs) could be suspected. We performed a cross-sectional study of 741 forensic autopsies undertaken at our institution in non-drug addicts aged 15 years or above from 2017 to 2019. We defined a high risk of miQTP by one detected QT-PM in a concentration above therapeutic level, or two or more detected QT-PMs in post mortem blood. We reviewed the autopsy reports from cases with a high miQTP-risk to identify cases with no other apparent cause of death. We discarded suicides and cases with lethal levels of QT-PMs. We identified 167 cases (22.5%) with high risk of miQTP, and discarded 36 suicides (4.9%) and 7 (0.9%) with lethal levels of QT-PMs. Apart from a high risk of miQTP, no other apparent explanation of the cause of death was present in seven (0.9%). In 18 cases (2.4%) with high miQTP-risk, the cause of death was primarily attributed to cardiac changes other than acute cardiovascular events. In conclusion, 22.5% had a high risk of miQTP, and fatal cardiac arrhythmia related to treatment with QT-PMs could be suspected in 0.9%. However, a genetic pro-arrhythmic background could not be excluded in our study. Furthermore, it is possible that QT-PMs could have played a role in some of the 2.4% of cases where the cause of death was mainly attributed to cardiac changes and the risk of miQTP was high.

Research Sun, 01 Aug 2021 17:51:40 +0200 8bd2be24-2b26-4078-9e8d-ee3a10914151
<![CDATA[Determination of camostat and its metabolites in human plasma – Preservation of samples and quantification by a validated UHPLC-MS/ MS method]]> Sørensen, L. K., Hasselstrøm, J. B., Damsgaard Gunst, J., Søgaard, O. S., Kjolby, M. Objectives: Camostat mesilate is a drug that is being repurposed for new applications such as that against COVID-19 and prostate cancer. This induces a need for the development of an analytical method for the quantification of camostat and its metabolites in plasma samples. Camostat is, however, very unstable in whole blood and plasma due to its two ester bonds. The molecule is readily hydrolysed by esterases to 4-(4-guanidinobenzoyloxy)phenylacetic acid (GBPA) and further to 4-guanidinobenzoic acid (GBA). For reliable quantification of camostat, a technique is required that can instantly inhibit esterases when blood samples are collected. Design and methods: An ultra-high-performance liquid chromatography-tandem mass spectrometry method (UHPLC-ESI-MS/MS) using stable isotopically labelled analogues as internal standards was developed and validated. Different esterase inhibitors were tested for their ability to stop the hydrolysis of camostat ester bonds. Results: Both diisopropylfluorophosphate (DFP) and paraoxon were discovered as efficient inhibitors of camostat metabolism at 10 mM concentrations. No significant changes in camostat and GBPA concentrations were observed in fluoride-citrate-DFP/paraoxon-preserved plasma after 24 h of storage at room temperature or 4 months of storage at −20 °C and −80 °C. The lower limits of quantification were 0.1 ng/mL for camostat and GBPA and 0.2 ng/mL for GBA. The mean true extraction recoveries were greater than 90%. The relative intra-laboratory reproducibility standard deviations were at a maximum of 8% at concentrations of 1–800 ng/mL. The trueness expressed as the relative bias of the test results was within ±3% at concentrations of 1–800 ng/mL. Conclusions: A methodology was developed that preserves camostat and GBPA in plasma samples and provides accurate and sensitive quantification of camostat, GBPA and GBA by UHPLC-MS/MS.

Research Fri, 01 Oct 2021 17:51:40 +0200 3bb1fca8-9218-459b-a7dd-cacdd01cadc5
<![CDATA[Bone pain in multiple myeloma (Bpmm)—a protocol for a prospective, longitudinal, observational study]]> Diaz-Delcastillo, M., Andrews, R. E., Mandal, A., Andersen, T. L., Chantry, A. D., Heegaard, A. M. Multiple myeloma (MM) is a bone marrow neoplasia that causes bone pain in 70% patients. While preclinical models of MM have suggested that both nerve sprouting and nerve injury may be causative for the pain, there is a lack of clinical data. Thus, the primary aims of this clinical study are: (1) to provide a deep characterization of the subjective experience of pain and quality of life in MM patients; (2) to investigate disturbances in the bone innervation of MM patients. Secondary aims include exploring correlations between pain and serum inflammatory and bone turnover biomarkers. In a prospective, observational study ( NCT04273425), patients with suspected MM requiring a diagnostic iliac crest biopsy at Sheffield Teaching Hospital (UK) are invited to participate. Consenting patients answer seven standardized questionnaires assessing pain, quality of life and catastrophizing. Bone turnover biomarkers and inflammatory cytokines are measured in fasting serum samples, and bone innervation is evaluated in diagnostic biopsies. MM patients are invited to a follow-up upon completion of first line treatment. This will be the first deep characterization of pain in MM patients and its correlation with disturbances in bone innervation. Understanding how bone turnover and inflammation correlate to pain in MM is crucial to identify novel analgesic targets for this condition.

Research Thu, 01 Apr 2021 17:51:40 +0200 5fc0941b-ef02-4cc1-8b01-d6a14e933fbb
<![CDATA[Fatal pulmonary fibrosis]]> Schwensen, H. F., Borreschmidt, L. K., Storgaard, M., Redsted, S., Christensen, S., Madsen, L. B. There is growing evidence of histopathological changes in autopsied individuals infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2); however, data on histopathological changes in autopsied patients with eradicated COVID-19 are limited. We performed an autopsy on a Caucasian female in her 80s, who died due to severe, bilateral pulmonary fibrosis after eliminated SARS-CoV-2 infection. In addition, CT scans from 2 months before infection and from 6 days prior to death were compared. Comparison of the CT scans showed bilateral development of widespread fibrosis in previously healthy lungs. Microscopic examination showed different areas with acute and organising diffuse alveolar damage and fibrosis with honeycomb-like remodelling and bronchial metaplasia. We here report a unique autopsy case with development of widespread pulmonary fibrosis in a woman in her 80s with previous COVID-19 and no history of pulmonary illnesses.

Research Tue, 01 Jun 2021 17:51:40 +0200 e7bc7c96-c19e-41e6-8a31-3739e56ec787