Bioavailability of Single-Dose SUBA-Itraconazole Compared to Conventional Itraconazole under Fasted and Fed Conditions.

Conventional itraconazole (C-ITZ) suffers from absorption variability. SUBA-itraconazole (S-ITZ) is more bioavailable than C-ITZ at steady state in a fed condition, but there are no data comparing the two under a fasted state. An open-label, single-dose, randomized, bioequivalence study was performed comparing S-ITZ to C-ITZ capsules under fasted and fed conditions in healthy adults measuring itraconazole and hydroxyitraconazole plasma levels. This study demonstrated less variability of S-ITZ compared to C-ITZ capsules under fasted conditions.

Open-Label Crossover Oral Bioequivalence Pharmacokinetics Comparison for a 3-Day Loading Dose Regimen and 15-Day Steady-State Administration of SUBA-Itraconazole and Conventional Itraconazole Capsules in Healthy Adults.

Super bioavailability (SUBA) itraconazole (S-ITZ), which releases drug in the duodenum, and conventional itraconazole (C-ITZ), which releases drug in the stomach, were compared in two pharmacokinetic (PK) studies: a 3-day loading dose study and a 15-day steady-state administration study. These were crossover oral bioequivalence studies performed under fed conditions in healthy adult volunteers. In the loading dose study, C-ITZ (two doses of 100 mg each) and S-ITZ (two doses of 65 mg each) were administered three times daily for 3 days and once on day 4 (n = 15). For the steady-state administration study, C-ITZ (two doses of 100 mg each) and S-ITZ (two doses of 65 mg each) were administered twice daily for 14 days and a last dose was administered 30 min after a meal on day 15 (n = 16). Blood samples collected throughout both studies were analyzed for ITZ and hydroxy-ITZ (OH-ITZ) levels. Least-squares geometric means were used to compare the maximum peak concentration of drug after administration at steady state prior to administration of the subsequent dose (Cmax_ss), the minimum drug level after administration prior to the subsequent dose (Ctrough), and the area under the curve over the dosing interval (AUCtau) of each formulation. The ratios of itraconazole (ITZ) and OH-ITZ for S-ITZ to C-ITZ were between 107% and 118% in both studies for Cmax_ss, Ctrough, and AUCtau, which were within the U.S. FDA-required bioequivalence range of 80% to 125%. At the end of the steady-state administration study, 13 of 16 volunteers obtained higher mean ITZ blood Ctrough levels of >1,000 ng/ml when they were administered S-ITZ (81%) than when they were administered C-ITZ (44%). The study drugs were well tolerated in both studies, with similar adverse events (AEs). All treatment-emergent AEs resolved after study completion. One volunteer receiving C-ITZ discontinued due to a treatment-unrelated AE in the steady-state administration study. No serious AEs were reported. Total, trough, and peak ITZ and OH-ITZ exposures were similar between the two formulations. Therefore, SUBA-ITZ, which has 35% less drug than C-ITZ, was bioequivalent to C-ITZ in healthy adult volunteers and exhibited a safety profile similar to that of C-ITZ.

Exposure to Persistent Organic Pollutants in Australian Waterbirds.

There is growing worldwide recognition of the threat posed by persistent organic pollutants (POPs) to wildlife populations. We aimed to measure exposure levels to POPs in a Southern Hemisphere aquatic waterbird species, the nomadic gray teal (Anas gracilis), which is found across Australia. We collected wings from 39 ducks harvested by recreational hunters at two sites (one coastal, one inland) in Victoria, southeastern Australia, in 2021. We examined three groups of POPs: nine congeners of polychlorinated biphenyls (PCBs), 13 organochlorine pesticides (OCPs), and 12 polycyclic aromatic hydrocarbons (PAHs). The PCBs, OCPs, and PAHs were detected at quantifiable levels in 13%, 72%, and 100% of birds, respectively. Of the congeners we tested for in PCBs, OCPs, and PAHs, 33%, 38%, and 100% were detected at quantifiable levels, respectively. The highest levels of exposure to POPs that we found were to the PAH benzo[b]fluoranthene, occurring at a concentration range of 1.78 to 161.05 ng/g wet weight. There were some trends detected relating to differences between geographical sites, with higher levels of several PAHs at the coastal versus inland site. There were several strong, positive associations among PAHs found. We discuss potential sources for the POPs detected, including industrial and agricultural sources, and the likely role of large-scale forest fires in PAH levels. Our results confirm that while Australian waterbirds are exposed to a variety of POPs, exposure levels are currently relatively low. Additional future investigations are required to further characterize POPs within Australian waterbird species. Environ Toxicol Chem 2024;43:736-747. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

New and legacy persistent organic pollutants (POPs) in breeding seabirds from the East Antarctic.

Persistent organic pollutants (POPs) are pervasive and a significant threat to the environment worldwide. Yet, reports of POP levels in Antarctic seabirds based on blood are scarce, resulting in significant geographical gaps. Blood concentrations offer a snapshot of contamination within live populations, and have been used widely for Arctic and Northern Hemisphere seabird species but less so in Antarctica. This paper presents levels of legacy POPs (polychlorinated biphenyls (PCBs), organochlorine pesticides (OCPs) and polybrominated diphenyl ethers (PBDEs)) and novel brominated flame retardants (NBFRs) in the blood of five Antarctic seabird species breeding within Prydz Bay, East Antarctica. Legacy PCBs and OCPs were detected in all species sampled, with Adélie penguins showing comparatively high ∑PCB levels (61.1 ± 87.6 ng/g wet weight (ww)) compared to the four species of flying seabirds except the snow petrel (22.5 ± 15.5 ng/g ww), highlighting that legacy POPs are still present within Antarctic wildlife despite decades-long bans. Both PBDEs and NBFRs were detected in trace levels for all species and hexabromobenzene (HBB) was quantified in cape petrels (0.3 ± 0.2 ng/g ww) and snow petrels (0.2 ± 0.1 ng/g ww), comparable to concentrations found in Arctic seabirds. These results fill a significant data gap within the Antarctic region for POPs studies, representing a crucial step forward assessing the fate and impact of legacy POPs contamination in the Antarctic environment.

Persistent organic pollutant (POPs) concentrations from great-winged petrels nesting in Western Australia.

Marine animals that traverse coastal and offshore environments are potentially exposed to multiple sources of pollution. Baseline data of pollutant concentrations of these fauna are needed in remote areas as human populations grow and economic development increases because changes may affect local wildlife in unforeseen ways. Persistent organic pollutant (POPs) concentrations were quantified in an understudied seabird, the great-winged petrel (Pterodroma macroptera), that breeds in southern Western Australia. Organochlorine pesticides, polychlorinated biphenyls (PCBs), and novel brominated flame retardants (NBFRs) were measured in adults. Total POPs concentrations ranged 5.6-46.4 ng g-1 ww. The most frequently detected POPs were the dichlorodiethyltrichloroethane (DDT) metabolite 4,4'DDE, the PCB CB-28, and the BFR polybrominated diphenyl ether BDE-99. These results contribute to the limited POPs data in marine fauna in this remote region, and the Southern Hemisphere, adding to the growing body of evidence that remote regions are affected by global trends of POPs distributions.

Adélie penguin colonies as indicators of brominated flame retardants (BFRs) in East Antarctica.

While persistent organic pollutant (POP) contamination within Antarctica is largely caused by long-range atmospheric transport (LRAT), Antarctic research bases have been shown to be local sources of POPs such as brominated flame retardants (BFRs). This study compared concentrations of seven polybrominated diphenyl ethers (PBDE) congeners and five novel flame retardants (NBFRs) found in Adélie penguin (Pygoscelis adeliae) colony soils near the Australian research stations, Mawson and Davis, to assess the stations as local sources of these contaminants and provide a much needed baseline for contamination of BFRs in East Antarctica. Soil samples (n = 46) were collected from Adélie colonies at close proximity to the research stations as well as further afield during the 2016-17 austral summer. Samples were analysed using selective pressurised liquid extraction (S-PLE) and gas chromatography coupled to tandem mass spectrometry (GC-MS/MS). PBDEs (BDE-28, -47, -99, -100, -153, -154 and -183) were detected in 45/46 samples with ∑7PBDE concentrations ranging from <0.01 to 1.63 ng/g dry weight (dw) and NBFRs (2,3,4,5,6-pentabromotoluene (PBT), 2,3,4,5,6-pentabromoethylbenzene (PBEB), hexabromobenzene (HBB), 2-ethylhexyl-2,3,4,5-tetrabromobenzoate (EH-TBB) and bis(2,4,6-tribromophenoxy) ethane (BTBPE)) detected in 20/46 samples, with a range of ∑5NBFR from not detected (ND) to 0.16 ng/g dw. Soils taken from around the Davis and Mawson research stations were more highly contaminated (n = 10) than penguin colonies (n = 27) and control areas not affiliated with breeding seabirds (n = 8). The most common congener detected was BDE-99, reflecting inputs from LRAT. However, the congener profiles of station soils supported the hypothesis that research stations are a local source of PBDEs to the Antarctic environment. In addition, the NBFR pentabromoethylbenzene (PBEB) was quantified for the first time in Antarctic soils, providing essential information for baseline contamination within the region and highlighting the need for ongoing monitoring as global regulations for the use of BFRs continuously change.

A baseline for POPs contamination in Australian seabirds: little penguins vs. short-tailed shearwaters.

While globally distributed throughout the world's ecosystems, there is little baseline information on persistent organic pollutants (POPs) in marine environments in Australia and, more broadly, the Southern Hemisphere. To fill this knowledge gap, we collected baseline information on POPs in migratory short-tailed shearwaters (Ardenna tenuirostris) from Fisher Island, Tasmania, and resident little penguins (Eudyptula minor) from Phillip Island, Victoria. Levels of polychlorinated biphenyls (PCBs), organochlorine pesticides (OCPs) and brominated flame retardants (BFRs) were determined from blood samples, with total contamination ranging 7.6-47.7 ng/g ww for short-tailed shearwaters and 0.12-46.9 ng/g ww for little penguins. In both species contamination followed the same pattern where PCBs>OCPs>BFRs. BFR levels included the presence of the novel flame retardant hexabromobenzene (HBB). These novel results of POPs in seabirds in southeast Australia provide important information on the local (penguins) and global (shearwaters) distribution of POPs in the marine environment.

CTLA-4 Blockade, during HIV Virus-Like Particles Immunization, Alters HIV-Specific B-Cell Responses.

Studies have shown that blockade of CTLA-4 promoted the expansion of germinal center B-cells in viral infection or immunization with model antigens. Few studies have evaluated the immunological consequences of CTLA-4 blockade during immunization against relevant vaccine candidates. Here, we investigated the effects of CTLA-4 blockade on HIV virus-like particles (VLPs) vaccination in a C57BL/6J mouse model. We found that CTLA-4 blockade during HIV VLP immunization resulted in increased CD4+ T-cell activation, promoted the expansion of HIV envelope (Env)-specific follicular helper T cell (Tfh) cells, and significantly increased HIV Gag- and Env-specific IgG with higher avidity and antibody-dependent cellular cytotoxicity (ADCC) capabilities. Furthermore, after only a single immunization, CTLA-4 blockade accelerated T-cell dependent IgG class switching and the induction of significantly high serum levels of the B-cell survival factor, A proliferation-inducing ligand (APRIL). Although no significant increase in neutralizing antibodies was observed, increased levels of class-switched Env- and Gag-specific IgG are indicative of increased polyclonal B-cell activation, which demonstrated the ability to mediate and enhance ADCC in this study. Altogether, our findings show that CTLA-4 blockade can increase the levels of HIV antigen-specific B-cell and antigen-specific Tfh cell activity and impact humoral immune responses when combined with a clinically relevant HIV VLP-based vaccine.

The psychoactive pollutant fluoxetine compromises antipredator behaviour in fish.

Pharmaceuticals are increasingly being detected in aquatic ecosystems worldwide. Particularly concerning are pharmaceutical pollutants that can adversely impact exposed wildlife, even at extremely low concentrations. One such contaminant is the widely prescribed antidepressant fluoxetine, which can disrupt neurotransmission and behavioural pathways in wildlife. Despite this, relatively limited research has addressed the behavioural impacts of fluoxetine at ecologically realistic exposure concentrations. Here, we show that 28-day fluoxetine exposure at two ecologically relevant dosages-one representing low surface water concentrations and another representing high effluent flow concentrations-alters antipredator behaviour in Eastern mosquitofish (Gambusia holbrooki). We found that fluoxetine exposure at the lower dosage resulted in increased activity levels irrespective of the presence or absence of a predatory dragonfly nymph (Hemianax papuensis). Additionally, irrespective of exposure concentration, fluoxetine-exposed fish entered the predator 'strike zone' more rapidly. In a separate experiment, fluoxetine exposure reduced mosquitofish freezing behaviour-a common antipredator strategy-following a simulated predator strike, although, in females, this reduction in behaviour was seen only at the lower dosage. Together, our findings suggest that fluoxetine can cause both non-monotonic and sex-dependent shifts in behaviour. Further, they demonstrate that exposure to fluoxetine at environmentally realistic concentrations can alter antipredator behaviour, with important repercussions for organismal fitness.

Toll-like receptor 3 adjuvant in combination with virus-like particles elicit a humoral response against HIV.

Human Immunodeficiency Virus (HIV) Virus-Like Particles (VLPs) composed of HIVIIIB Gag and HIVBaL gp120/gp41 envelope are a pseudovirion vaccine capable of presenting antigens in their native conformations. To enhance the immunogenicity of the HIV Env antigen, VLPs were coupled to VesiVax Conjugatable Adjuvant Lipid Vesicles (CALV) containing one of four toll-like-receptor (TLR) ligands, each activating a receptor with distinct cellular localization and downstream pathways. C57BL/6 mice were vaccinated by intranasal prime followed by two sub-cheek boosts and their sera immunoglobulin and neutralizing potency were measured over a duration of 3months after vaccination. PBS control, VLPs alone, CALV+VLPs, and VLPs complexed with CALV and ligands for TLR2 (PAM3CAG), TLR3 (dsRNA), TLR4 (MPLA), or TLR7/8 (resiquimod) were evaluated based on antibody titer, IgG1 and IgG2c class switching, germinal center formation, T follicular cells and potency of neutralizing antibodies. Consistently, the TLR3 ligand dsRNA complexed to CALV and in combination with VLPs (CALV(dsRNA)+VLPs) induced the strongest response. CALV(dsRNA)+VLPs induced the highest titers against the recombinant vaccine antigens clade B Bal gp120 and pr55 Gag. Additionally, CALV(dsRNA)+VLPs induced cross-clade antibodies, represented by high titers of antibody to clade c 96ZM651 gp120. CALV(dsRNA)+VLPs induced predominantly IgG2c over IgG1, a response associated with T helper type 1 (Th1)-like cytokines. In turn, CALV(dsRNA)+VLP immunized mice generated the most potent neutralizing antibodies against HIV strain MN.3. Finally, at time of sacrifice, a significant increase in germinal center B cells and T follicular cells was detected in mice which received CALV(dsRNA)+VLPs compared to PBS. Our results indicate that CALV(dsRNA) is a superior adjuvant for HIV VLPs in generating a Th1-like immunoglobulin profile, while prolonging lymph node germinal centers, T follicular cells and generating neutralizing antibodies to a highly sensitive tier 1A variant of HIV.

A Novel Prime and Boost Regimen of HIV Virus-Like Particles with TLR4 Adjuvant MPLA Induces Th1 Oriented Immune Responses against HIV.

HIV virus-like particles (VLPs) present the HIV envelope protein in its native conformation, providing an ideal vaccine antigen. To enhance the immunogenicity of the VLP vaccine, we sought to improve upon two components; the route of administration and the additional adjuvant. Using HIV VLPs, we evaluated sub-cheek as a novel route of vaccine administration when combined with other conventional routes of immunization. Of five combinations of distinct prime and boost sequences, which included sub-cheek, intranasal, and intradermal routes of administration, intranasal prime and sub-cheek boost (IN+SC) resulted in the highest HIV-specific IgG titers among the groups tested. Using the IN+SC regimen we tested the adjuvant VesiVax Conjugatable Adjuvant Lipid Vesicles (CALV) + monophosphoryl lipid A (MPLA) at MPLA concentrations of 0, 7.5, 12.5, and 25 µg/dose in combination with our VLPs. Mice that received 12.5 or 25 µg/dose MPLA had the highest concentrations of Env-specific IgG2c (20.7 and 18.4 µg/ml respectively), which represents a Th1 type of immune response in C57BL/6 mice. This was in sharp contrast to mice which received 0 or 7.5 µg MPLA adjuvant (6.05 and 5.68 µg/ml of IgG2c respectively). In contrast to IgG2c, MPLA had minor effects on Env-specific IgG1; therefore, 12.5 and 25 µg/dose of MPLA induced the optimal IgG1/IgG2c ratio of 1.3. Additionally, the percentage of germinal center B cells increased significantly from 15.4% in the control group to 31.9% in the CALV + 25 µg MPLA group. These mice also had significantly more IL-2 and less IL-4 Env-specific CD8+ T cells than controls, correlating with an increased percentage of Env-specific central memory CD4+ and CD8+ T cells. Our study shows the strong potential of IN+SC as an efficacious route of administration and the effectiveness of VLPs combined with MPLA adjuvant to induce Env specific Th1-oriented HIV-specific immune responses.