18F-fluoro-deoxyglucose (FDG) is currently widely used for PET/CT imaging of large vessel vasculitis (LVV) in giant cell arteritis (GCA), but its performance is influenced by glucocorticoid therapy. We aimed to compare 68Ga-HA-DOTATATE (DOTA), a somatostatin-analog tracer specific for macrophages, to FDG for detection of LVV using PET/CT in a small number of patients with active GCA. In this pilot study, 8 active GCA patients (with disease confirmation by either temporal artery biopsy or positive large vessel imaging) who had initiated glucocorticoids within 2 weeks of enrollment were prospectively consented and sequentially scanned with both FDG PET/CT and 68Ga-HA-DOTATATE PET/CT. Images were reviewed by 2 experienced Nuclear Medicine physicians. Tracer uptake was assessed semi-quantitatively in 8 vascular territories (thoracic and abdominal aorta, bilateral subclavian, vertebral, carotid, femoral/iliac, temporal arteries and coronary arteries) using SUVmax. Target-background ratios (TBR) were calculated using both right atrium (RA) mean (TBRRA) and liver mean (TBRliver) as comparator. Index vessel was defined as the vessel with the greatest tracer uptake in each participant. SUVmax and TBR’s of individual vascular territories and index vessels were compared using students’ t tests. P<0.05 was considered statistically significant. The median patient age was 71.5 years (range 64-82), and 4 (50%) were women. See Table 1 for baseline characteristics (Table 1). Five of 8 (62.5%) FDG scans had grade 2 visual uptake in the large vessels, consistent with active LVV. Tracer uptake could be detected semi-quantitatively in the medium-large arteries in both FDG and DOTA scans. FDG scans demonstrated a significantly higher RA activity than DOTA (RA SUVmean 1.88 vs 0.36, p<0.001), while DOTA had a significantly higher liver uptake (liver SUVmean 7.54 vs 2.39, p<0.001). Vascular uptake (as measured by both SUVmax and TBRliver) was significantly higher in FDG than DOTA scans in every vascular territory (p<=0.05 for all comparisons), including index vessels (SUVmax 4.04 vs 1.91, p=0.01, TBRliver 1.73 vs 0.27, p=0.0009). Table 1. Baseline demographics and clinical characteristics of GCA patients. 68Ga-HA-DOTATATE uptake can be detected in the aorta and medium branch vessels, including temporal and coronary arteries, of patients with active GCA receiving glucocorticoids. The distribution of DOTA uptake differs from that of FDG, with significantly lower blood pool uptake and higher liver uptake. In this pilot study, the arterial uptake of DOTA was overall lower than that of FDG, suggesting this may not be an optimal substitute for the more established tracer.

[18F]Fluorodeoxyglucose (FDG) is widely used in PET/CT imaging to detect large vessel vasculitis in giant cell arteritis (GCA), but its performance is suboptimal in patients receiving glucocorticoids. We aimed to compare [68Ga]Ga-HA-DOTA-TATE, a somatostatin 2-analogue tracer, to [18F]FDG in a pilot study of patients with GCA. Eight patients with active GCA were prospectively, sequentially scanned with both [18F]FDG PET/CT and [68Ga]Ga-HA-DOTA-TATE PET/CT imaging. Images were evaluated by 2 blinded nuclear medicine specialists. Tracer uptake was assessed in 8 vascular territories using SUVmax, and target-background ratios (TBR) were calculated using both right atrium (TBRRA) and liver mean (TBRliver). Mean SUVmax and TBR of individual vascular territories and index vessels were compared. The patient median age was 71.5 years (range 64–82), and 4 (50 https://clinicaltrials.gov/search?cond=dotatate .

Catheter-associated urinary tract infection is one of the most common healthcare-associated infections, with biofilm formation playing a key role in its pathogenesis. Indwelling medical devices introduce ideal pathways inside the body for pathogens and feature surfaces conducive to biofilm development, often leading to severe clinical infections recalcitrant to antimicrobials. When bacteria and fungi switch to biofilm mode of growth, they produce a matrix in the form of extracellular polymeric substances (EPS). This creates a unique environment for growing virulent colonisers and persisting cells while forming a shielding barrier against immune system attacks, antimicrobial agents and mechanical removal by fluid shear forces. To address this challenge, LubriShieldTM - a novel permanent coating - was invented and evenly applied to both internal and external surfaces of indwelling urinary Foley catheters. Without releasing active substances, it effectively prevented pathogens from producing biofilm. The superhydrophilic coating, incorporating a proprietary anti-fouling ligand, significantly inhibited colonising uropathogens from forming biofilm for up to 14 days in artificial urine medium without microbial killing (up to 99% reduction, P < 0.001). In a glass bladder flow model, LubriShieldTM still significantly reduced biofilm formation by 83% (P < 0.0001). Importantly, LubriShield™ maintained its antibiofilm efficacy even after conditioning with fibrinogen, a host-derived protein known to promote bacterial attachment (P = 0.007). RNA-seq analysis revealed significant downregulation of genes associated with microbial EPS formation on the coated surfaces. Additionally, microorganisms adhering to LubriShieldTM coated catheters showed a 78% increased susceptibility to antibiotics compared to those on uncoated catheters (P = 0.004).

This article provides an overview of preclinical theranostic radiopharmaceutical development, highlighting aspects of the preclinical development stages that can lead towards a clinical trial. The key stages of theranostic radiopharmaceutical development are outlined, including target selection, tracer development, radiopharmaceutical synthesis, automation and quality control, in vitro radiopharmaceutical analysis, selecting a suitable in vivo model, preclinical imaging and pharmacokinetic analysis, preclinical therapeutic analysis, dosimetry, toxicity, and preparing for clinical translation. Each stage is described and augmented with examples from the literature. Finally, an outlook on the prospects for the radiopharmaceutical theranostics field is provided.

Background Nuclear medicine has made enormous progress in the past decades. However, there are still significant inequalities in patient access among different countries, which could be mitigated by improving access to and availability of radiopharmaceuticals. Main body This paper summarises major considerations for a suitable pharmaceutical regulatory framework to facilitate patient access to radiopharmaceuticals. These include the distinct characteristics of radiopharmaceuticals which require dedicated regulations, considering the impact of the variable complexity of radiopharmaceutical preparation, personnel requirements, manufacturing practices and quality assurance, regulatory authority interfaces, communication and training, as well as marketing authorisation procedures to ensure availability of radiopharmaceuticals. Finally, domestic and regional supply to ensure patient access via alternative regulatory pathways, including in-house production of radiopharmaceuticals, is described, and an outlook on regulatory challenges faced by new developments, such as the use of alpha emitters, is provided. Conclusions All these considerations are an outcome of a dedicated Technical Meeting organised by the IAEA in 2023 and represent the views and opinions of experts in the field, not those of any regulatory authorities.

Poor nutritional status is common among human immunodeficiency virus (HIV)-infected patients including vitamin D (vitD3) deficiency. We conducted a double-blinded, randomized, and placebo-controlled trial in Addis Ababa, Ethiopia, to investigate if daily nutritional supplementation with vitD3 (5000 IU) and phenylbutyrate (PBA, 2 × 500 mg) could mediate beneficial effects in treatment-naïve HIV patients. Primary endpoint: the change in plasma HIV-1 comparing week 0 to 16 using modified intention-to-treat (mITT, n = 197) and per-protocol (n = 173) analyses. Secondary endpoints: longitudinal HIV viral load, T cell counts, body mass index (BMI), middle-upper-arm circumference (MUAC), and 25(OH)D3 levels in plasma. Baseline characteristics were detectable viral loads (median 7897 copies/mL), low CD4+ (median 410 cells/µL), and elevated CD8+ (median 930 cells/µL) T cell counts. Most subjects were vitD3 deficient at enrolment, but a gradual and significant improvement of vitD3 status was demonstrated in the vitD3 + PBA group compared with placebo (p < 0.0001) from week 0 to 16 (median 37.5 versus 115.5 nmol/L). No significant changes in HIV viral load, CD4+ or CD8+ T cell counts, BMI or MUAC could be detected. Clinical adverse events were similar in both groups. Daily vitD3 + PBA for 16 weeks was well-tolerated and effectively improved vitD3 status but did not reduce viral load, restore peripheral T cell counts or improve BMI or MUAC in HIV patients with slow progressive disease. Clinicaltrials.gov NCT01702974.

Dysbiosis and a dysregulated gut immune barrier function contributes to chronic immune activation in HIV-1 infection. We investigated if nutritional supplementation with vitamin D and phenylbutyrate could improve gut-derived inflammation, selected microbial metabolites, and composition of the gut microbiota. Treatment-naïve HIV-1-infected individuals (n = 167) were included from a double-blind, randomized, and placebo-controlled trial of daily 5000 IU vitamin D and 500 mg phenylbutyrate for 16 weeks (Clinicaltrials.gov NCT01702974). Baseline and per-protocol plasma samples at week 16 were analysed for soluble CD14, the antimicrobial peptide LL-37, kynurenine/tryptophan-ratio, TMAO, choline, and betaine. Assessment of the gut microbiota involved 16S rRNA gene sequencing of colonic biopsies. Vitamin D + phenylbutyrate treatment significantly increased 25-hydroxyvitamin D levels (p < 0.001) but had no effects on sCD14, the kynurenine/tryptophan-ratio, TMAO, or choline levels. Subgroup-analyses of vitamin D insufficient subjects demonstrated a significant increase of LL-37 in the treatment group (p = 0.02), whereas treatment failed to significantly impact LL-37-levels in multiple regression analysis. Further, no effects on the microbiota was found in number of operational taxonomic units (p = 0.71), Shannon microbial diversity index (p = 0.82), or in principal component analyses (p = 0.83). Nutritional supplementation with vitamin D + phenylbutyrate did not modulate gut-derived inflammatory markers or microbial composition in treatment-naïve HIV-1 individuals with active viral replication.

•This review summarizes the recent development of positron emission tomography (PET) radioligands for γ-aminobutyric acid A (GABAA) receptors.•Clinical and translational applications of GABA PET radioligands are highlighted.•The importance and challenge of radioligands with increased sensitivity to efficiently measure GABA release are also discussed.•We conclude that PET tracers with increased GABA sensitivity will have broad applications across several brain disorders.

The escape of anti-self B cells from tolerance mechanisms like clonal deletion, receptor editing, and anergy results in the production of autoantibodies, which is a hallmark of many autoimmune disorders. In this study, we demonstrate that both germline sequences and somatic mutations contribute to autospecificity of B cell clones. For this issue, we investigated the development of antinuclear autoantibodies (ANAs) and their repertoire in two different mouse models. First, in aging mice that were shown to gain several autoimmune features over time including ANAs. Second, in mice undergoing a chronic graft-versus-host disease (GVHD), thereby developing systemic lupus erythematosus-like symptoms. Detailed repertoire analysis revealed that somatic hypermutations (SHM) were present in all Vh and practically all Vl regions of ANAs generated in these two models. The ANA B cell repertoire in aging mice was restricted, dominated by clonally related Vh1-26/Vk4-74 antibodies. In the collection of GVHD-derived ANAs, the repertoire was less restricted, but the usage of the Vh1-26/Vk4-74 combination was still apparent. Germline conversion showed that the SHM in the 4-74 light chain are deterministic for autoreactivity. Detailed analysis revealed that antinuclear reactivity of these antibodies could be induced by a single amino acid substitution in the CDR1 of the Vk4-74. In both aging B6 and young GVHD mice, conversion of the somatic mutations in the Vh and Vl regions of non Vh1-26/Vk4-74 using antibodies showed that B cells with a germline-encoded V gene could also contribute to the ANA-reactive B cell repertoire. These findings indicate that two distinct pathways generate ANA-producing B cells in both model systems. In one pathway, they are generated by Vh1-26/Vk4-74 expressing B cells in the course of immune responses to an antigen that is neither a nuclear antigen nor any other self-antigen. In the other pathway, ANA-producing B cells are derived from progenitors in the bone marrow that express B cell receptors (BCRs), which bind to nuclear antigens and that escape tolerance induction, possibly as a result of crosslinking of their BCRs by multivalent determinants of nuclear antigens.

Interleukin-7 (IL-7) and Flt3-ligand (FL) are two cytokines important for the generation of B cells, as manifested by the impaired B cell development in mice deficient for either cytokine or their respective receptors and by the complete block in B cell differentiation in the absence of both cytokines. IL-7 is an important survival and proliferation factor for B cell progenitors, whereas FL acts on several early developmental stages, prior to B cell commitment. We have generated mice constitutively over-expressing both IL-7 and FL. These double transgenic mice develop splenomegaly and lymphadenopathy characterized by tremendously enlarged lymph nodes even in young animals. Lymphoid, myeloid and dendritic cell numbers are increased compared to mice over-expressing either of the two cytokines alone and the effect on their expansion is synergistic, rather than additive. B cell progenitors, early progenitors with myeloid and lymphoid potential (EPLM), common lymphoid progenitors (CLP) and lineage-, Sca1+, kit+ (LSK) cells are all increased not only in the bone marrow but also in peripheral blood, spleen and even lymph nodes. When transplanted into irradiated wild-type mice, lymph node cells show long-term multilineage reconstitution, further confirming the presence of functional hematopoietic progenitors therein. Our double transgenic mouse model shows that sustained and combined over-expression of IL-7 and FL leads to a massive expansion of most bone marrow hematopoietic progenitors and to their associated presence in peripheral lymphoid organs where they reside and potentially differentiate further, thus leading to the synergistic increase in mature lymphoid and myeloid cell numbers. The present study provides further in vivo evidence for the concerted action of IL-7 and FL on lymphopoiesis and suggests that extramedullary niches, including those in lymph nodes, can support the survival and maintenance of hematopoietic progenitors that under physiological conditions develop exclusively in the bone marrow.

Low vitamin D (vitD3) is one of the most common nutritional deficiencies in the world known to be associated with numerous medical conditions including infections such as tuberculosis (TB). In this study, vitD3 status and its association with the antimicrobial peptide, human cathelicidin (LL-37), was investigated in Ethiopian patients with different clinical forms of TB. Patients with active TB (n = 77) and non-TB controls (n = 78) were enrolled in Ethiopia, while another group of non-TB controls (n = 62) was from Sweden. Active TB included pulmonary TB (n = 32), pleural TB (n = 20), and lymph node TB (n = 25). Concentrations of 25-hydroxyvitamin D3 (25(OH)D3) were assessed in plasma, while LL-37 mRNA was measured in peripheral blood and in samples obtained from the site of infection. Median 25(OH)D3 plasma levels in active TB patients were similar to Ethiopian non-TB controls (38.5 versus 35.0 nmol/L) and vitD3 deficiency (<50 nmol/L) was common in both groups (73%). Ethiopians (low latitude) had significantly lower 25(OH)D3 levels compared with Swedish non-TB controls (51.0 nmol/L, high latitude), but vitD3 status was not affected by tuberculin-positivity or HIV infection. Patients with local lymph node TB had significantly higher 25(OH)D3 levels compared with pulmonary TB patients (48.0 versus 29.0 nmol/L). Moreover, plasma 25(OH)D3 levels correlated with local LL-37 expression in granulomatous lesions in TB infected lymph nodes. Instead, systemic LL-37 mRNA expression in blood cells was elevated compared with the site of infection in pulmonary and pleural TB. Low vitD3 status may be associated with an enhanced peripheral expression of LL-37 in patients with intrathoracic TB that could result from chronic inflammation.

This paper presents the irradiation and processing of high-current Mo-100 targets at the University of Alberta (UofA) in a GMP compliant setting. For purpose of comparison with a second production facility, additional studies at Centre Hospitalier Universitaire de Sherbrooke (CHUS) are also described. Introduction: More than 70% of today's diagnostic radiopharmaceuticals are based on Tc-99m, however the conventional supply chain for obtaining Tc-99m is fragile. The aim of this work was to demonstrate reliable high yield production and processing of Tc-99m with medium-energy, high-current, cyclotrons. Methods: We used two cyclotrons (TR-24, Advanced Cyclotron Systems, Inc) for irradiations with 22 MeV or 24 MeV incident energy and 400 mu A current up to a maximum of 6 h. The irradiated Mo-100 was dissolved using peroxide, basified using ammonium carbonate, and purified using a PEG-based solid phase extraction technique. Results: High-yield productions with 22 MeV (400 mu A, 6 h) yielded an average isolated [Tc-99m]TcO4- yield of 878 GBq +/- 99 GBq (23.7 Ci +/- 2.7 Ci) decay corrected to EOB, n = 8 (isolated saturation yield: 4.36 +/- 0.49 GBq/mu A). Irradiations with 24 MeV (400 mu A, 6 h) resulted in an average isolated [Tc-99m]TcO4- yield of 993 GBq +/- 100 GBq (26.8 Ci +/- 2.7 Ci) decay corrected to EOB, n = 7 (isolated saturation yield: 4.97 +/- 0.50 GBq/mu A). These yields corresponds to 600-700 GBq (16-19 Ci) of [Tc-99m]TcO4- at release (i.e. 3 hour post-EOB). For all tested batches, the QC results were within the recently published specifications in the European Pharmacopoeia. Conclusion: Reliable near-TBq production yields for Tc-99m can be obtained using medium-energy cyclotrons. This work presents evidence that medium-energy high-current cyclotrons can provide high yields of [Tc-99m]TcO4- with radionuclidic impurities levels within the specifications of the existing European Pharmacopoeia monograph, indicating that this technology can have a share in the future Tc-99m supply market. (C) 2018 Elsevier Inc. All rights reserved.

Background: Novel vaccine strategies are required to provide protective immunity in tuberculosis (TB) and prevent development of active disease. We investigated the safety and immunogenicity of a novel TB vaccine candidate, H4:IC31 (AERAS-404) that is composed of a fusion protein of M. tuberculosis antigens Ag85B and TB10.4 combined with an IC31 (R) adjuvant.Methods: BCG-vaccinated healthy subjects were immunized with various antigen (5, 15, 50, 150 mu g) and adjuvant (0,100, 500 nmol) doses of the H4:IC31 vaccine (n = 106) or placebo (n = 18) in two randomized, double-blind, placebo-controlled phase I studies conducted in a low TB endemic setting in Sweden and Finland. The subjects were followed for adverse events and CD4(+) T cell responses.Results: H4:IC31 vaccination was well tolerated with a safety profile consisting of mostly mild to moderate self-limited injection site pain, myalgia, arthralgia, fever and post-vaccination inflammatory reaction at the screening tuberculin skin test injection site. The H4:IC31 vaccine elicited antigen-specific CD4(+) T cell proliferation and cytokine production that persisted 18 weeks after the last vaccination. CD4(+) T cell expansion, IFN-gamma production and multifunctional CD4(+) Th1 responses were most prominent after two doses of H4:IC31 containing 5, 15, or 50 mu g of H4 in combination with the 500 nmol IC31 adjuvant dose.Conclusions: The novel TB vaccine candidate, H4:IC31, demonstrated an acceptable safety profile and was immunogenic, capable of triggering multifunctional CD4+ T cell responses in previously BCG-vaccinated healthy individuals. These dose-escalation trials provided evidence that the optimal antigen-adjuvant dose combinations are 5, 15, or 50 mu g of H4 and 500 nmol of IC31. (C) 2017 The Authors. Published by Elsevier Ltd.

Objectives: The 6-exon-spanning ‘canonical’ Interleukin-7 (IL-7c) is a non-redundant cytokine in human T-cell homeostasis that undergoes extensive alternative pre-mRNA splicing. The IL-7 gene variant lacking, exon 5 (IL-7d5), exhibits agonistic effects as compared to IL-7c. We studied in this report for the first time the protein expression of IL-7d5 variant in tissues and its role in monocyte activation. Methods: We visualized the expression of IL-7d5 protein by immunohistochemistry in both healthy and malignant (human) tissues and investigated the impact of IL-7d5 stimulation on CD14+ monocytes using gene expression analysis and flow cytometry. Results: IL-7d5 is largely expressed by human epithelial cells, yet also by stromal cells in malignant lesions. Gene expression analysis in CD14+ monocytes, induced by the 6-exon spanning IL-7 or IL7d5 showed similar changes resulting in a pro-inflammatory phenotype and increased expression of genes involved in lipid metabolism. IL7d5 was superior in inducing upregulation of the oxidised low density lipoprotein receptor (OLR), measured by flow cytometry, in CD14+ cells. Conclusion: IL-7d5, produced from non-transformed and transformed cells, may contribute to chronic inflammatory responses and development of ‘foamy’ cells by increased OLR1 expression that mediates increased oxLDL uptake. © 2017 Published by Elsevier Ltd on behalf of International Society for Infectious Diseases. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

ObjectivesThe 6-exon-spanning 'canonical' Interleukin-7 (IL-7c) is a non-redundant cytokine in human T-cell homeostasis that undergoes extensive alternative pre-mRNA splicing. The IL-7 gene variant lacking, exon 5 (IL-7δ5), exhibits agonistic effects as compared to IL-7c. We studied in this report for the first time the protein expression of IL-7δ5 variant in tissues and its role in monocyte activation.MethodsWe visualized the expression of IL-7δ5 protein by immunohistochemistry in both healthy and malignant (human) tissues and investigated the impact of IL-7δ5 stimulation on CD14+ monocytes using gene expression analysis and flow cytometry.ResultsIL-7δ5 is largely expressed by human epithelial cells, yet also by stromal cells in malignant lesions. Gene expression analysis in CD14+ monocytes, induced by the 6-exon spanning IL-7 or IL-7δ5 showed similar changes resulting in a pro-inflammatory phenotype and increased expression of genes involved in lipid metabolism. IL7δ5 was superior in inducing upregulation of the oxidised low density lipoprotein receptor (OLR), measured by flow cytometry, in CD14+ cells.ConclusionIL-7δ5, produced from non-transformed and transformed cells, may contribute to chronic inflammatory responses and development of 'foamy' cells by increased OLR1 expression that mediates increased oxLDL uptake.