Determination of ivermectin in plasma and whole blood using LC-MS/MS.

Background: Ivermectin is a widely used drug for the treatment of helminthiasis and filariasis worldwide, and it has also shown promise for malaria elimination through its potent mosquito-lethal activity. The objective of this study was to develop and validate a high-throughput and sensitive method to quantify ivermectin in plasma and whole blood samples, using automated sample extraction followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Methods: Phospholipids were removed in patient whole blood (100 µl) and plasma (100 µl) samples using a 96-well plate Hybrid-solid phase extraction technique. Ivermectin and its isotope-labelled internal standard (ivermectin-D2) were separated on an Agilent Poroshell 120 EC-C18 50mm × 3.0mm I.D. 2.7µm, using a mobile phase of acetonitrile: ammonium formate 2 mM containing 0.5% formic acid (90: 10, v/v). Detection was performed using a triple quadrupole mass spectrometer in the positive ionization mode. Results: The method was validated in the concentration range 0.970 - 384 ng/ml in both plasma and whole blood matrices. Intra- and inter-batch precisions during the validation were below 15%. There was no carryover or matrix effects detected. Ivermectin is a stable compound and results showed no degradation in the different stability tests. Conclusions: The validated method proved to have high sensitivity and precision, good selectivity and to be suitable for clinical application or laboratory quantification of ivermectin in plasma or whole blood samples.

Cost-effectiveness of mass drug administration for control of scabies in Ethiopia: a decision-analytic model.

Background: The strategies to control scabies in highly endemic populations include individual case/household management and mass drug administration (MDA). We used a decision-analytic model to compare ivermectin-based MDA and individual case/household management (referred to as "usual care") for control of scabies in Ethiopia at different prevalence thresholds for commencing MDA. Methods: A decision-analytic model was based on a repeated population survey conducted in Northern Ethiopia in 2018-2020, which aimed to evaluate the secondary impact of single-dose ivermectin MDA for the control of onchocerciasis on scabies prevalence. The model estimates the number of scabies cases and costs of two treatment strategies (MDA and usual care) based on their effectiveness, population size, scabies prevalence, compliance with MDA, medication cost, and other parameters. Results: In the base-case analysis with a population of 100,000 and scabies prevalence of 15%, the MDA strategy was both more effective and less costly than usual care. The probability of MDA being cost-effective at the current cost-effectiveness threshold (equivalent to the cost of usual care) was 85%. One-way sensitivity analyses showed that the MDA strategy remained dominant (less costly and more effective) in 22 out of 26 scenarios. MDA was not cost-effective at scabies prevalence <10%, MDA effectiveness <85% and population size <5,000. An increase in the cost of ivermectin from 0 (donated) to 0.54 US$/dose resulted in a decrease in the probability of MDA being cost-effective from 85% to 17%. At 0.25 US$/dose, the MDA strategy was no longer cost-effective. Conclusions: The model provides robust estimates of the costs and outcomes of MDA and usual care and can be used by decision-makers for planning and implementing scabies control programmes. Results of our analysis suggest that single-dose ivermectin MDA is cost-effective in scabies control and can be initiated at a scabies prevalence >10%.

Epidemiology of epilepsy in Wulu County, an onchocerciasis-endemic area in South Sudan.

Background: We sought to investigate the epidemiology of epilepsy in Wulu County (Lakes State, South Sudan), and document the onchocerciasis transmission status in the study villages. Methods: In February 2024, a community-based epilepsy study was conducted Wulu County and participants were surveyed via a door-to-door approach in five villages, namely: Kombi, Makundi Center, Tonjo, War-Pac, and Woko. All village residents were asked about ivermectin intake during the 2023 round of community-directed treatment with ivermectin (CDTI). In addition, children aged 3-9 years were tested for Ov16 antibodies using a rapid diagnostic test. Epilepsy diagnosis in screened individuals was confirmed by a physician. Results: We surveyed 1355 persons in the five study sites. The overall CDTI coverage in 2023 was 67.4 %. Fifty-five persons with epilepsy (PWE) were identified (prevalence 4.1 %) and a history of nodding seizures was noted in 11/55 (20 %) PWE. The mean age of PWE was 21.5 ± 9.6 years, with 32 (58.2 %) being males. Epilepsy onset frequently occurred under 5 years of age (38.6 % of cases). In two PWE, seizure onset occurred during the past 12 months (annual incidence: 147.6 per 100,000 persons). Twenty-nine PWE (52.7 %) were taking anti-seizure medicines, but only five were taking them daily. Overall, Ov16 seroprevalence in children aged 3-9 years (n = 119) was 15.1 % and differed across villages, peaking at 30.9 % in Woko village where epilepsy prevalence was also highest (7.1 %). Of the 35 recorded deaths during the past two years, 9 (25.7 %) occurred in PWE. Annual estimates for epilepsy mortality and fatality rates were 323.7 per 100,000 persons and 7031.3 per 100,000 PWE, respectively. Conclusion: High epilepsy prevalence was found in Wulu, particularly in villages with persistent onchocerciasis transmission. Frequent epilepsy onset among under-fives suggests that perinatal/early childhood etiologies are common. Appropriate measures should be instituted to prevent and treat epilepsy in Wulu villages.

Cutaneous larva migrans, a neglected tropical disease (NTD) of skin, with severe secondary eczematization at an unusual location.

Cutaneous larva migrans (CLM), commonly referred to as creeping eruption, is an infectious condition caused by various types of hookworms. It primarily affects the superficial layers of the skin owing to the absence of hyaluronidases and proteases. Typically, the distal lower extremities are the most commonly affected areas. The presence of distinctive lesions characterized by erythematous, winding, or serpentine tracks, slightly elevated from the skin surface, is indicative of the condition. Diagnosis primarily relies on clinical observation. Dermoscopy reveals multiple segmented yellowish-linear regions corresponding to pustules along the larval path. Treatment typically involves the use of topical and oral ivermectin, oral albendazole, and topical thiabendazole cream.

The Use of Systemically Absorbed Drugs to Explore An In Vitro Bioequivalence Approach For Comparing Non-Systemically Absorbed Active Pharmaceutical Ingredients in Drug Products For Use in Dogs.

PURPOSE: Currently, for veterinary oral formulations containing one or more active pharmaceutical ingredient (API) that are not systemically absorbed and act locally within the gastrointestinal (GI) tract, the use of terminal clinical endpoint bioequivalence (BE) studies is the only option for evaluating product BE. This investigation explored the use of a totality of evidence approach as an alternative to these terminal studies. METHODS: Three formulations of tablets containing ivermectin plus praziquantel were manufactured to exhibit distinctly different in vitro release characteristics. Because these APIs are highly permeable, plasma drug concentrations served as a biomarker of in vivo dissolution. Tablets were administered to 27 healthy Beagle dogs (3-way crossover) and the rate and extent of exposure of each API for each formulation was compared in a pairwise manner. These results were compared to product relative in vitro dissolution profiles in 3 media. In vivo and in vitro BE predictions were compared. RESULTS: In vivo/in vitro inconsistencies in product relative performance were observed with both compounds when considering product performance across the 3 dissolution media. Formulation comparisons flagged major differences that could explain this outcome. CONCLUSIONS: The finding of an inconsistent in vivo/in vitro relationship confirmed that in vitro dissolution alone cannot assure product BE for veterinary locally acting GI products. However, when combined with a comparison of product composition and manufacturing method, this totality of evidence approach can successfully alert scientists to potential therapeutic inequivalence, thereby supporting FDA's efforts to Replace, Reduce, and/or Refine terminal animal studies.

Escalating Threat of Drug-Resistant Human Scabies: Current Insights and Future Directions.

Background: Scabies is a prevalent dermatological condition with significant public health implications. The recent rise in drug-resistant scabies presents new challenges for effective disease management and control. Methods: A comprehensive literature review was conducted using PubMed, Cochrane Library, and Web of Science. Studies published from 2000 to August 2024 were considered, focusing on those reporting drug-resistant scabies and advancements in treatment approaches. Results: Clinical studies, in vitro investigations, and case reports show significant resistance of human scabies to permethrin. Main resistance mechanisms involve genetic mutations in the mites' voltage-gated sodium channels (VGSCs) and enhanced activity or expression of the detoxifying enzyme glutathione S-transferase (GST). Resistance to ivermectin and benzyle benzoate, although suggested by some authors, seems less obvious. The clinical evidence of widespread ivermectin resistance in human scabies infestations is lacking, despite indications of increased tolerance in laboratory settings and anecdotal reports of resistance in patients with crusted scabies. Benzyl benzoate resistance in scabies mites remains unconfirmed. Conclusions: Permethrin-resistant scabies is an escalating threat requiring new management strategies and updated guidelines. Infection control measures, alternative treatments, and ongoing research into new therapeutics are crucial to mitigate the impact of drug-resistant scabies.

Mosquitocidal efficacy and pharmacokinetics of single-dose ivermectin versus three-day dose regimen for malaria vector control compared with albendazole and no treatment: An open-label randomized controlled trial.

OBJECTIVES: When malaria vectors consume ivermectin in a blood meal, their survival probability decreases, potentially reducing malaria transmission during mass drug administrations. However, questions remain regarding the optimal dosing. This study aimed to compare the mosquitocidal effect and pharmacokinetics of two-dose regimens of ivermectin for malaria vector control. DESIGN: We conducted an open-label randomized control trial in Kenya, staggered in blocks with sequential intervention groups and parallel controls. Participants were randomly assigned (2:1:1:1) using computer random-sequence generation, unstratified, with one block of six pharmacokinetics-only participants (single-dose ivermectin) and six blocks of four participants (3:1 intervention vs control), to receive single-dose ivermectin (400 mcg/kg, n = 12), three daily doses (3-day regimen 300 mcg/kg, n = 6), albendazole (400 mg, n = 6), or no treatment (negative control, n = 6). Our primary outcome was Anopheles gambiae survival (time-to-event [days]) after blood feeding up to 10 days after drug administration. We also evaluated pharmacokinetics (peak plasma and capillary blood concentration, areas under the plasma and capillary blood concentration-time curve from time of last administration to time of last observation, time to reach peak plasma and capillary blood concentration, terminal elimination half-life) up to 7 days after treatment. RESULTS: A total of 36 healthy volunteers aged 21-32 years were recruited into the study and followed up to completion, with two participants not attending the visit on day 28. All drug regimens were well-tolerated. Both regimens showed significant mosquitocidal effect in the first 7 days. At 10 days after treatment, the single dose presented superior longevity of effect (adjusted hazard ratio = 3.91; 95% confidence interval = 1.93-7.93; P <0.001) compared with the triple dose (adjusted hazard ratio = 1.79; 95% confidence interval = 0.88-3.62; P = 0.0.11). Albendazole had, overall, no mosquitocidal effect. CONCLUSIONS: It is unclear why a single dose led to increased bio-efficacy compared with a triple dose. We recommend trials investigating ivermectin mass drug administrations for malaria control to consider single-dose ivermectin. A single-dose regimen is also expected to present additional operational advantages compared with a 3-day regimen, leading to improved programmatic suitability.

Revitalizing common drugs for antibacterial, quorum quenching, and antivirulence potential against Pseudomonas aeruginosa: in vitro and in silico insights.

In the post-antibiotic era, antivirulence therapies are becoming refractory to the clinical application of existing antimicrobial regimens. Moreover, in an attempt to explore alternate intervention strategies, drug repurposing is gaining attention over development of novel drugs/antimicrobials. With the prevalence of multidrug resistance and high medical burden associated with Pseudomonas aeruginosa, there is an urgent need to devise novel therapeutics to combat this bacterial pathogen. In this context, the present study was undertaken to scrutinize the anti-quorum sensing (QS) and antivirulence potential of commonly consumed drugs such as fexofenadine (FeX), ivermectin (IvM), nitrofurantoin (NiT), levocetrizine (LvC), atorvastatin (AtS), and aceclofenac (AcF), against P. aeruginosa. The methodology involved assessment of antibacterial activity against P. aeruginosa PAO1 and quorum quenching (QQ) potential using Agrobacterium tumefaciens NTL4 biosensor strain. The antivirulence prospects were investigated by estimating the production of hallmark virulence factors in P. aeruginosa accompanied by molecular docking to predict drug associations with the QS receptors. Interestingly, all the drugs harbored antibacterial, anti-QS, and antivirulence potential in vitro, which consequently disrupted QS circuits and attenuated pseudomonal virulence phenotypically by significantly lowering the production of pyocyanin, hemolysin, pyochelin, and total bacterial protease in vitro. Moreover, the findings were validated by computational studies that predicted strong molecular interactions between the test drugs and QS receptors of P. aeruginosa. Hence, this study is the first to suggest the prospect of repurposing FeX, IvM, NiT, LvC, AtS, and AcF against P. aeruginosa.

A comparative field efficacy trial of three treatment programs against endo- and ectoparasites in naturally infected dogs.

Introduction: Tropical climates in remote Aboriginal and Torres Strait Islander communities in northern Australia are conducive to the transmission of canine helminths such as hookworms, as well as ectoparasites such as fleas and ticks. In addition to their veterinary importance, these parasites may present a zoonotic risk either directly, or as potential vectors for bacterial pathogens. These factors necessitate efficacious and effective antiparasitic treatment programs for community dogs. Methods: A cluster-randomised trial was performed on three islands in the Torres Strait to examine the short-term efficacy and medium-term effectiveness of three treatment programs. Treatments administered included oral oxibendazole/praziquantel (Paragard®) and oral afoxolaner (Nexgard®); topical moxidectin/imidacloprid (Advocate®) and imidacloprid/flumethrin collars (Seresto®); and off-label oral ivermectin (Bomectin®). Canine faecal samples were collected and examined for endoparasites by faecal flotation and real-time PCR at baseline, 7-11 days after treatment and 6 months later. Results: The proportion of dogs positive for Ancylostoma caninum at baseline and negative at day 7-11 was 9% (95% CI 4.4-17.4) for dogs treated with oxibendazole, 56.4% (95% CI 41-70.7) for moxidectin, and 89.7% (95% CI 73.6-96.4) for ivermectin. Faecal flotation results showed a greater than 90% egg reduction in 29.2% (95% CI 19.9-40.5) of dogs treated with oxibendazole, 79.4% (95% CI 63.2-89.7) for moxidectin, and 95% (95% CI 76.4-99.1) for off-label ivermectin. Elimination of ectoparasite infestation was observed at day 7-11 in 69.9% (95% CI 56.7-80.1) of dogs treated with afoxolaner, 80% (95% CI 60.9-91.1) with imidacloprid/flumethrin collars, and 0% (95% CI 0-11.7) for off-label ivermectin. Mixed effects modelling revealed only treatment group to be significantly associated with outcome measures. Discussion: Based on these study results, the poor efficacy of oxibendazole against A. caninum renders it inept for treatment, while ivermectin and moxidectin were suitable. Ivermectin was unsuitable for ectoparasite treatment due to its poor efficacy, while afoxolaner and imidacloprid/flumethrin collars appear suitable.