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Abstract Background Huntington’s disease (HD) is a rare neurodegenerative disorder caused by an expansion of the CAG trinucleotide repeat in the huntingtin gene which encodes the mutant huntingtin protein (mHTT) that is associated with HD-related neuropathophysiology. Noninvasive visualization of mHTT aggregates in the brain, with positron emission tomography (PET), will allow to reliably evaluate the efficacy of therapeutic interventions in HD. This study aimed to assess the radiation burden of [18F]CHDI-650, a novel fluorinated mHTT radioligand, in humans based on both in vivo and ex vivo biodistribution in mice and subsequent determination of dosimetry for dosing in humans. Results Wild-type male and female CD-1 Swiss mice (n = 15/sex) were used to assess in vivo PET imaging-based and ex vivo biodistribution-based tracer distribution of [18F]CHDI-650 at 30-, 60-, 120-, 240- and 360-min post-injection. Three-dimensional volumes of interest of the organs were drawn on the co-registered PET/CT image and organs were collected after dissection. Organ radioactivity levels were determined using both modalities. The residence time was calculated and extrapolated to human phantoms. The absorbed and effective doses were computed with OLINDA/EXM 2.2 and IDAC-Dose2.1. Ex vivo and PET-imaging biodistribution of [18F]CHDI-650 showed rapid washout after 30 min in most of the organs with the highest uptake in the gallbladder and urine in mice. Extrapolation of the data to human phantoms with OLINDA showed a total mean in vivo based effective dose of 21.7 μSv/MBq with the highest equivalent organ dose in the urinary bladder wall (4.52 μSv/MBq). The total mean ex vivo based effective dose was calculated to be 20.6 μSv/MBq. The highest equivalent organ dose ex vivo in the urinary bladder wall was estimated to be 4.22 μSv/MBq. The predicted exposure in humans using IDAC-Dose correlated well to those obtained with OLINDA for both in vivo and ex vivo measurements (r = 0.9320 and r = 0.9368, respectively). Conclusions Dosimetry analysis indicated absorbed and effective doses of [18F]CHDI-650 are well below the recommended limits, suggesting that the radioligand is suitable for clinical assessment. Based on the highest effective dose estimates, an injection of 370 MBq in humans would result in a radiation dose of 8.03 mSv.

Abstract The melanocortin receptor 4 (MC4R) belongs to the melanocortin receptor family of G-protein coupled receptors and is a key switch in the leptin-melanocortin molecular axis that controls hunger and satiety. Brain-produced hormones such as α-melanocyte-stimulating hormone (agonist) and agouti-related peptide (inverse agonist) regulate the molecular communication of the MC4R axis but are promiscuous for melanocortin receptor subtypes and induce a wide array of biological effects. Here, we use a chimeric construct of conformation-selective, nanobody-based binding domain (a ConfoBody Cb80) and active state-stabilized MC4R-β2AR hybrid for efficient de novo discovery of a sequence diverse panel of MC4R-specific, potent and full agonistic nanobodies. We solve the active state MC4R structure in complex with the full agonistic nanobody pN162 at 3.4 Å resolution. The structure shows a distinct interaction with pN162 binding deeply in the orthosteric pocket. MC4R peptide agonists, such as the marketed setmelanotide, lack receptor selectivity and show off-target effects. In contrast, the agonistic nanobody is highly specific and hence can be a more suitable agent for anti-obesity therapeutic intervention via MC4R.

Abstract BackgroundData quality is fundamental to maintaining the trust and reliability of health data for both primary and secondary purposes. However, before the secondary use of health data, it is essential to assess the quality at the source and to develop systematic methods for the assessment of important data quality dimensions. ObjectiveThis case study aims to offer a dual aim—to assess the data quality of height and weight measurements across 7 Belgian hospitals, focusing on the dimensions of completeness and consistency, and to outline the obstacles these hospitals face in sharing and improving data quality standards. MethodsFocusing on data quality dimensions completeness and consistency, this study examined height and weight data collected from 2021 to 2022 within 3 distinct departments—surgical, geriatrics, and pediatrics—in each of the 7 hospitals. ResultsVariability was observed in the completeness scores for height across hospitals and departments, especially within surgical and geriatric wards. In contrast, weight data uniformly achieved high completeness scores. Notably, the consistency of height and weight data recording was uniformly high across all departments. ConclusionsA collective collaboration among Belgian hospitals, transcending network affiliations, was formed to conduct this data quality assessment. This study demonstrates the potential for improving data quality across health care organizations by sharing knowledge and good practices, establishing a foundation for future, similar research.

Abstract Background Kinetic modeling in positron emission tomography (PET) requires measurement of the tracer plasma activity in the absence of a suitable reference region. To avoid invasive blood sampling, the use of an image derived input function has been proposed. However, an accurate delineation of the blood pool region in the PET image is necessary to obtain unbiased blood activity. Here, to perform brain kinetic modeling in [18F]SynVesT-1 dynamic scans, we make use of non-negative matrix factorization (NMF) to unmix the activity signal from the different tissues that can contribute to the heart region activity, and extract only the left ventricle activity in an unbiased way. This method was implemented in dynamic [18F]SynVesT-1 scans of mice anesthetized with either isoflurane or ketamine–xylazine, two anesthestics that we showed to affect differently radiotracer kinetics. The left ventricle activity (NMF-IDIF) and a manually delineated cardiac activity (IDIF) were compared with arterial blood samples (ABS), and for isoflurane anesthetized mice, arteriovenous (AV) shunt blood data were compared as well. Finally, brain regional 2 tissue compartment modeling was performed using IDIF and NMF-IDIF, and the model fit accuracy (weighted symmetrical mean absolute percentage error, wsMAPE) as well as the total volume of distribution (V T) were compared. Results In isoflurane anesthetized mice, the difference between ABS and NMF-IDIF activity (+ 12.8 $$\pm$$ ± 11%, p = 0.0023) was smaller than with IDIF (+ 16.4 $$\pm$$ ± 9.8%, p = 0.0008). For ketamine–xylazine anesthetized mice the reduction in difference was larger (NMF-IDIF: 16.9 $$\pm$$ ± 10%, p = 0.0057, IDIF: 56.3 $$\pm$$ ± 14%, p

ABSTRACTIn vitro reverse transcription of full-length HIV-1 RNA extracted from the blood plasma of people living with HIV-1 remains challenging. Here, we describe the initiation of reverse transcription of plasma-derived viral RNA in the absence of an exogenous primer. Real-time PCR and Sanger sequencing were applied to identify the source and to monitor the outcome of this reaction. Results demonstrated that during purification of viral RNA from plasma, tRNA(Lys-3) is co-extracted in a complex with the viral RNA. In the presence of a reverse transcription enzyme, this tRNA(Lys-3) can induce reverse transcription, a reaction that is not confined to transcription of the 5’ end of the viral RNA. A range of cDNA products is generated, most of them indicative for the occurrence of in vitro strand transfer events that involve translocation of cDNA from the 5’ end to random positions on the viral RNA. This process results in the formation of cDNAs with large internal deletions. However, near full-length cDNA and cDNA with sequence patterns resembling multiple spliced HIV-1 RNA were also detected. Despite its potential to introduce significant bias in the interpretation of results across various applications, tRNA(Lys-3)-driven reverse transcription has been overlooked thus far. A more in-depth study of this tRNA-driven in vitro reaction may provide new insight into the complex process of in vivo HIV-1 replication.IMPORTANCEThe use of silica-based extraction methods for purifying HIV-1 RNA from viral particles is a common practice, but it involves co-extraction of human tRNA(Lys-3) due to the strong interactions between these molecules. This co-extraction becomes particularly significant when the extracted RNA is used in reverse transcription reactions, as the tRNA(Lys-3) then serves as a primer. Reverse transcription from tRNA(Lys-3) is not confined to cDNA synthesis of the 5’ end of the viral RNA but extends across various regions of the viral genome through in vitro strand transfer events. Co-extraction of tRNA(Lys-3) has been overlooked thus far, despite its potential to introduce bias in downstream, reverse transcription-related applications. The observed events in the tRNA(Lys-3)-induced in vitro reverse transcription resemble in vivo replication processes. Therefore, these reactions may offer a unique model to better understand the replication dynamics of HIV-1.

Hava Izci,1 Gilles Macq,2 Tim Tambuyzer,2 Harlinde De Schutter,2 Hans Wildiers,1,3 Francois P Duhoux,4 Evandro de Azambuja,5 Donatienne Taylor,6 Gracienne Staelens,7 Guy Orye,8 Zuzana Hlavata,9 Helga Hellemans,10 Carine De Rop,11 Patrick Neven,1,3 Freija Verdoodt2 1KU Leuven - University of Leuven, Department of Oncology, Leuven, B-3000, Belgium; 2Belgian Cancer Registry, Research Department, Brussels, Belgium; 3University Hospitals Leuven, Multidisciplinary Breast Center, Leuven, B-3000, Belgium; 4Department of Medical Oncology, King Albert II Cancer Institute, Cliniques Universitaires Saint-Luc, Brussels, Belgium; 5Institut Jules Bordet and l’Université Libre de Bruxelles (U.L.B), Brussels, Belgium; 6CHU UCL Namur, Site Sainte-Elisabeth, Namur, Belgium; 7Multidisciplinary Breast Center, General Hospital Groeninge, Kortrijk, Belgium; 8Department of Obstetrics and Gynecology, Jessa Hospital, Hasselt, Belgium; 9Department of Medical Oncology, CHR Mons-Hainaut, Mons, Hainaut, Belgium; 10Department of Obstetrics and Gynaecology, AZ Delta, Roeselaere, Belgium; 11Department of Obstetrics and Gynaecology, Imelda Hospital, Bonheiden, BelgiumCorrespondence: Hava Izci, KU Leuven, Department of oncology, Herestraat 49 Box 7003-06, Leuven, 3000, Belgium, Email hava.izci@kuleuven.bePurpose: High-quality population-based cancer recurrence data are scarcely available, mainly due to complexity and cost of registration. For the first time in Belgium, we developed a tool to estimate distant recurrence after a breast cancer diagnosis at the population level, based on real-world cancer registration and administrative data.Methods: Data on distant cancer recurrence (including progression) from patients diagnosed with breast cancer between 2009– 2014 were collected from medical files at 9 Belgian centers to train, test and externally validate an algorithm (i.e., gold standard). Distant recurrence was defined as the occurrence of distant metastases between 120 days and within 10 years after the primary diagnosis, with follow-up until December 31, 2018. Data from the gold standard were linked to population-based data from the Belgian Cancer Registry (BCR) and administrative data sources. Potential features to detect recurrences in administrative data were defined based on expert opinion from breast oncologists, and subsequently selected using bootstrap aggregation. Based on the selected features, classification and regression tree (CART) analysis was performed to construct an algorithm for classifying patients as having a distant recurrence or not.Results: A total of 2507 patients were included of whom 216 had a distant recurrence in the clinical data set. The performance of the algorithm showed sensitivity of 79.5% (95% CI 68.8– 87.8%), positive predictive value (PPV) of 79.5% (95% CI 68.8– 87.8%), and accuracy of 96.7% (95% CI 95.4– 97.7%). The external validation resulted in a sensitivity of 84.1% (95% CI 74.4– 91.3%), PPV of 84.1% (95% CI 74.4– 91.3%), and an accuracy of 96.8% (95% CI 95.4– 97.9%).Conclusion: Our algorithm detected distant breast cancer recurrences with an overall good accuracy of 96.8% for patients with breast cancer, as observed in the first multi-centric external validation exercise.Keywords: machine learning, breast cancer, distant metastases, recurrences, algorithm, administrative data

Detection dogs were trained to detect SARS-CoV-2 infection based on armpit sweat odor. Sweat samples were collected using cotton pads under the armpits of negative and positive human patients, confirmed by qPCR, for periods of 15–30 min. Multiple hospitals and organizations throughout Belgium participated in this study. The sweat samples were stored at −20°C prior to being used for training purposes. Six dogs were trained under controlled atmosphere conditions for 2–3 months. After training, a 7-day validation period was conducted to assess the dogs’ performances. The detection dogs exhibited an overall sensitivity of 81%, specificity of 98%, and an accuracy of 95%. After validation, training continued for 3 months, during which the dogs’ performances remained the same. Gas chromatography/mass spectrometry (GC/MS) analysis revealed a unique sweat scent associated with SARS-CoV-2 positive sweat samples. This scent consisted of a wide variety of volatiles, including breakdown compounds of antiviral fatty acids, skin proteins and neurotransmitters/hormones. An acceptability survey conducted in Belgium demonstrated an overall high acceptability and enthusiasm toward the use of detection dogs for SARS-CoV-2 detection. Compared to qPCR and previous canine studies, the detection dogs have good performances in detecting SARS-CoV-2 infection in humans, using frozen sweat samples from the armpits. As a result, they can be used as an accurate pre-screening tool in various field settings alongside the PCR test.

Industrial chicory (Cichorium intybus var. sativum) is a biannual crop mostly cultivated for extraction of inulin, a fructose polymer used as a dietary fiber. F1 hybrid breeding is a promising breeding strategy in chicory but relies on stable male sterile lines to prevent self-pollination. Here, we report the assembly and annotation of a new industrial chicory reference genome. Additionally, we performed RNA-Seq on subsequent stages of flower bud development of a fertile line and two cytoplasmic male sterile (CMS) clones. Comparison of fertile and CMS flower bud transcriptomes combined with morphological microscopic analysis of anthers, provided a molecular understanding of anther development and identified key genes in a range of underlying processes, including tapetum development, sink establishment, pollen wall development and anther dehiscence. We also described the role of phytohormones in the regulation of these processes under normal fertile flower bud development. In parallel, we evaluated which processes are disturbed in CMS clones and could contribute to the male sterile phenotype. Taken together, this study provides a state-of-the-art industrial chicory reference genome, an annotated and curated candidate gene set related to anther development and male sterility as well as a detailed molecular timetable of flower bud development in fertile and CMS lines.

Electron Beam (EB) irradiation was utilized to decontaminate model systems of industrial polymers that contain a brominated flame retardant (BFR). Acrylonitrile-butadiene-styrene (ABS) and Polycarbonate (PC) are two types of polymers commonly found in Waste Electrical and Electronic Equipment (WEEE). In this study, these polymers were exposed to EB irradiation to degrade DecaBromoDiphenylEther (DBDE), one of the most toxic BFRs. Fourier-transform infrared spectroscopy analysis demonstrated an 87% degradation rate of DBDE for the ABS-DBDE system and 91% for the PC-DBDE system following an 1800 kGy irradiation dose. Thermal analysis using Differential Scanning Calorimetry revealed the presence of crosslinking in ABS and a minor reduction in the glass transition temperature of PC after EB processing. Polymers exhibited thermal stability after photolysis, as indicated by thermogravimetric analysis. In summary, EB irradiation had no impact on the overall thermal properties of both polymers. High-resolution mass spectrometry analysis has confirmed the debromination of both ABS-DBDE and PC-DBDE systems. Therefore, the results obtained are promising and could offer an alternative approach for removing bromine and other additives from plastic E-waste.

Background: Huntington's disease (HD) is a rare neurodegenerative disorder caused by an expansion of the CAG trinucleotide repeat in the huntingtin gene which encodes the mutant huntingtin protein (mHTT) that is associated with HD-related neuropathophysiology. Noninvasive visualization of mHTT aggregates in the brain, with positron emission tomography (PET), will allow to reliably evaluate the efficacy of therapeutic interventions in HD. This study aimed to assess the radiation burden of [18F]CHDI-650, a novel fluorinated mHTT radioligand, in humans based on both in vivo and ex vivo biodistribution in mice and subsequent determination of dosimetry for dosing in humans. Results: Wild-type male and female CD-1 Swiss mice (n = 15/sex) were used to assess in vivo PET imaging-based and ex vivo biodistribution-based tracer distribution of [18F]CHDI-650 at 30-, 60-, 120-, 240- and 360-min post-injection. Three-dimensional volumes of interest of the organs were drawn on the co-registered PET/CT image and organs were collected after dissection. Organ radioactivity levels were determined using both modalities. The residence time was calculated and extrapolated to human phantoms. The absorbed and effective doses were computed with OLINDA/EXM 2.2 and IDAC-Dose2.1. Ex vivo and PET-imaging biodistribution of [18F]CHDI-650 showed rapid washout after 30 min in most of the organs with the highest uptake in the gallbladder and urine in mice. Extrapolation of the data to human phantoms with OLINDA showed a total mean in vivo based effective dose of 21.7 μSv/MBq with the highest equivalent organ dose in the urinary bladder wall (4.52 μSv/MBq). The total mean ex vivo based effective dose was calculated to be 20.6 μSv/MBq. The highest equivalent organ dose ex vivo in the urinary bladder wall was estimated to be 4.22 μSv/MBq. The predicted exposure in humans using IDAC-Dose correlated well to those obtained with OLINDA for both in vivo and ex vivo measurements (r = 0.9320 and r = 0.9368, respectively). Conclusions: Dosimetry analysis indicated absorbed and effective doses of [18F]CHDI-650 are well below the recommended limits, suggesting that the radioligand is suitable for clinical assessment. Based on the highest effective dose estimates, an injection of 370 MBq in humans would result in a radiation dose of 8.03 mSv. [ABSTRACT FROM AUTHOR]

Purpose: Positron emission tomography (PET) imaging of mutant huntingtin (mHTT) aggregates is a potential tool to monitor disease progression as well as the efficacy of candidate therapeutic interventions for Huntington's disease (HD). To date, the focus has been mainly on the investigation of 11C radioligands; however, favourable 18F radiotracers will facilitate future clinical translation. This work aimed at characterising the novel [18F]CHDI-650 PET radiotracer using a combination of in vivo and in vitro approaches in a mouse model of HD. Methods: After characterising [18F]CHDI-650 using in vitro autoradiography, we assessed in vivo plasma and brain radiotracer stability as well as kinetics through dynamic PET imaging in the heterozygous (HET) zQ175DN mouse model of HD and wild-type (WT) littermates at 9 months of age. Additionally, we performed a head-to-head comparison study at 3 months with the previously published [11C]CHDI-180R radioligand. Results: Plasma and brain radiometabolite profiles indicated a suitable metabolic profile for in vivo imaging of [18F]CHDI-650. Both in vitro autoradiography and in vivo [18F]CHDI-650 PET imaging at 9 months of age demonstrated a significant genotype effect (p < 0.0001) despite the poor test–retest reliability. [18F]CHDI-650 PET imaging at 3 months of age displayed higher differentiation between genotypes when compared to [11C]CHDI-180R. Conclusion: Overall, [18F]CHDI-650 allows for discrimination between HET and WT zQ175DN mice at 9 and 3 months of age. [18F]CHDI-650 represents the first suitable 18F radioligand to image mHTT aggregates in mice and its clinical evaluation is underway. [ABSTRACT FROM AUTHOR]