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•First to evaluate mucosal inflammation at five weeks post anthelmintic treatment.•Mucosal inflammation varied based upon larvicidal treatment.•Mucosal inflammation varied between 2 ...and 5 weeks post anthelmintic treatment.•Goblet Cell hyperplasia was increased at 5 weeks post treatment.
Members of Cyathostominae are pervasive parasites of equids that can cause larval cyathostominosis, a potentially life-threatening disease that occurs when a multitude of encysted larvae synchronously excyst from the wall of the large intestine. Moxidectin and fenbendazole are the two current labeled drugs that target the encysted larval stages; however, there is limited knowledge of the local inflammatory response to the larvae and to the two treatments in clinically healthy horses. This study is the first to evaluate the local inflammatory response to cyathostomin larvae and to larvicidal treatment at 2 and 5 weeks post treatment. Thirty-six ponies with naturally acquired cyathostomin infections were randomly allocated into 3 groups: Group 1, fenbendazole at 10 mg/kg for 5 days, Group 2, a single dose of moxidectin at 0.4 mg/kg, and Group 3, untreated controls. Tissue samples from the cecum and dorsal and ventral colons were used for histopathological and immunohistochemical evaluation. Tissues were stained with routine hematoxylin and eosin (H&E) for light microscopy and immunohistochemically for MAC387, CD20, and CD3 for differentiation of activated macrophages, B cells, and T cells, respectively. Semiquantitative scores were assigned for all inflammatory cell types and fibrous connective tissue. Larvae observed by light microscopy were enumerated and classified by stage. Mucosal ulcerations and submucosal granulomas were also enumerated. Mean macrophage scores were higher in the moxidectin group than the fenbendazole group (p = 0.0185) and the control group had a higher activated macrophage score than both treatment groups (p = 0.0104, p = 0.0004). T lymphocyte scores were higher in the moxidectin group when compared to the control group (p = 0.0069). Goblet cell hyperplasia scores were elevated at 5 weeks post treatment compared to 2 weeks post treatment (p = 0.0047) and were elevated in the ventral colon compared to the dorsal colon (p = 0.0301). Eosinophil scores were elevated surrounding degenerative larvae when compared to intact larvae (p = 0.0001). Mucosal ulcerations were found only in the control group at 2 weeks post treatment. This study found subtle inflammatory differences between treatment groups but provided new information about goblet cells and eosinophils in relation to encysted cyathostomin larvae.
•Fill-FLOTAC homogenizer yielded higher strongylid egg count accuracy.•Fill-FLOTAC did not affect egg count precision.•Mini-FLOTAC disc associated with higher precision.•McMaster counting slide ...associated with higher accuracy.
Recommendations for control of equine strongylid parasites are based on regular determination of fecal egg counts to identify high strongylid shedders and to evaluate treatment efficacy. The McMaster technique has long been used as the standard egg counting technique in equine veterinary practice in most parts of the world, but recent work has found the Mini-FLOTAC technique to perform with significantly better accuracy and precision. The Mini-FLOTAC system comes with a homogenizing device, termed the Fill-FLOTAC, and it has been hypothesized that this device might have a significant impact on accuracy and precision. The aim of the present study was to investigate the impact of the Fill-FLOTAC homogenizer in comparison with the classical McMaster approach, where samples are suspended in flotation medium by stirring with tongue depressor in a plastic cup. The study compared the McMaster and Mini-FLOTAC techniques, but also included cross-over versions where the Fill-FLOTAC was used with the McMaster chamber, and the tongue depressor and plastic cup homogenizing method was used with the Mini-FLOTAC counting disc. Fecal samples were collected from horses naturally infected with mixed strongylid species. Five samples were included from each of the following egg count levels: 0–500, 501–1000, and >1000 eggs per gram (EPG). Each sample was then analyzed with all four set-ups with three subsamples collected from the same suspension, and three repeated counts determined on each subsample. Both the Fill-FLOTAC homogenizer (p = 0.0098) and the McMaster counting chamber (p = 0.0298) were significantly associated with higher strongylid egg counts, whereas the Mini-FLOTAC chamber was associated with a lower coefficient of variation (p < 0.0001). Precision, however, was not associated with homogenization method (p = 0.9341). Taken together, this study suggests that while the homogenizing method has a positive effect on egg count accuracy, the counting chamber appears to primarily affect precision.
Nanosecond pulsed electric fields (nsPEFs) applied to cells can induce different biological effects depending on pulse duration and field strength. One known process is the induction of apoptosis ...whereby nsPEFs are currently investigated as a novel cancer therapy. Another and probably related change is the breakdown of the cytoskeleton. We investigated the elasticity of rat liver epithelial cells WB-F344 in a monolayer using atomic force microscopy (AFM) with respect to the potential of cells to undergo malignant transformation or to develop a potential to metastasize. We found that the elastic modulus of the cells decreased significantly within the first 8 min after treatment with 20 pulses of 100 ns and with a field strength of 20 kV/cm but was still higher than the elasticity of their tumorigenic counterpart WB-ras. AFM measurements and immunofluorescent staining showed that the cellular actin cytoskeleton became reorganized within 5 min. However, both a colony formation assay and a cell migration assay revealed no significant changes after nsPEF treatment, implying that cells seem not to adopt malignant characteristics associated with metastasis formation despite the induced transient changes to elasticity and cytoskeleton that can be observed for up to 1 h.
BackgroundPatients with rheumatic disorders treated with TNF-α inhibitors have a 3–4 fold increased risk of developing TB. Our local population (Slough, UK) has a very high TB incidence with a rate ...of 58.5 per 100,000 population in 2012, approximately four times the national average. Latent TB in our population, many of whom are on immunomodulatory treatment, is thus likely to be several-fold higher.NICE guidance recommends that immunocompromised patients should be screened for latent TB using either an Interferon Gamma Release Assay (IGRA) alone or in combination with a tuberculin skin test.Two currently available IGRAs are the TB-ELISPOT and QuantiFERON-TB Gold In Tube (QFT) test. The NICE guidelines do not distinguish between these two assays. Whilst they have enhanced our ability to diagnose latent TB their use in immunosuppressed patients has not been widely evaluated. Furthermore, there is limited evidence to support the superiority of either of the two assays.ObjectivesWe aim to:Study the effectiveness and outcomes of our current TB screening programme prior to biologic therapy in our high risk immunosuppressed patientsCompare two currently used IGRAs (ELISPOT and QFT) in this population of patients.MethodsRetrospective data was obtained from our patient database. 246 consecutive patients (39% male and 61% female; age range 29 – 86 years; 78.5% Caucasian, 19.5% Asian and 2% other) were identified with inflammatory rheumatic disorders who had undergone clinical screening, a chest radiograph and an IGRA (either one or both of TB ELISPOT or QFT) prior to use of a biologic agent between 2008 and 2015.Results16/246 (6.5%) patients had a positive IGRA. Of these 15 received chemoprophylaxis for latent TB. 12 (75%) of these patients were Caucasian and the remainder were Asian. 230/246 (93.5%) patients were IGRA negative. 190 patients were receiving immunosuppressant therapy at the time of screening and 245 patients screened were subsequently treated with TNF inhibitors. In this 7-year period there have been no cases of TB reactivation reported to date.Of the patients screened with both IGRAs, the majority of patients tested negative for both (158/171, 92.4%). 13/171 (7.6%) patients had a positive result. Of these 6 tested positive for both ELISPOT and QFT, 4 had a positive ELISPOT only, and the remaining 3 had a positive QFT test only. The concordance of the two assays used was thus 95.9% (164/171 patients).ConclusionsThe absence of TB reactivation in our high risk immunosuppressed population receiving biologic therapy indicates the effectiveness of our current screening strategy.The current study demonstrates good concordance between the ELISPOT and QFT assays, suggesting that the use of either test on its own is sufficient for screening for latent TB in this population. This data supports current NICE guidance on diagnosing latent TB in the immunocompromised population.ReferencesPublic Health England. Tuberculosis in Thames Valley: Annual review (2013 data). https://www.gov.uk/government/uploads/system/uploads/attachment_data/file/397901/TBThamesValley2013data.pdfNational Institute of Health and Clinical Excellence. Tuberculosis - full guideline internet. NICE. 2016. http://www.nice.org.uk/guidance/ng33Ding T, Ledingham J et al. BSR and BHPR Rheumatoid Arthritis Guidelines on Safety of anti-TNF therapies. Rheumatology. 2010;49:2217–19Disclosure of InterestNone declared
Ascarid parasites infect a variety of hosts and regular anthelmintic treatment is recommended for all species. Parascaris spp. is the only ascarid species with widespread anthelmintic resistance, ...which allows for the study of resistance mechanisms. The purpose of this study was to establish an in vitro drug exposure protocol for adult anthelmintic-naïve Parascaris spp. and report a preliminary transcriptomic analysis in response to drug exposure. Live worms were harvested from foal necropsies and maintained in RPMI-1640 at 37 °C. Serial dilutions of oxibendazole (OBZ) and ivermectin (IVM) were prepared for in vitro drug exposure, and worm viability was monitored over time. In a second drug trial, worms were used for transcriptomic analysis. The final drug concentrations employed were OBZ at 40.1 μm (10 μg mL-1) and IVM at 1.1 μm (1 μg mL-1) for 24 and 3 h, respectively. The RNA-seq analysis revealed numerous differentially expressed genes, with some being potentially related to drug detoxification and regulatory mechanisms. This report provides a method for in vitro drug exposure and the phenotypic responses for Parascaris spp., which could be extrapolated to other ascarid parasites. Finally, it also provides preliminary transcriptomic data following drug exposure as a reference point for future studies of Parascaris spp.
Gamma-hydroxybutyrate (GHB; or sodium oxybate) is an endogenous GHB-/gamma-aminobutyric acid B receptor agonist. It is approved for application in narcolepsy and has been proposed for the potential ...treatment of Alzheimer's disease, Parkinson's disease, fibromyalgia, and depression, all of which involve neuro-immunological processes. Tryptophan catabolites (TRYCATs), the cortisol-awakening response (CAR), and brain-derived neurotrophic factor (BDNF) have been suggested as peripheral biomarkers of neuropsychiatric disorders. GHB has been shown to induce a delayed reduction of T helper and natural killer cell counts and alter basal cortisol levels, but GHB's effects on TRYCATs, CAR, and BDNF are unknown.
Therefore, TRYCAT and BDNF serum levels, as well as CAR and the affective state (Positive and Negative Affect Schedule PANAS) were measured in the morning after a single nocturnal dose of GHB (50 mg/kg body weight) in 20 healthy male volunteers in a placebo-controlled, balanced, randomized, double-blind, cross-over design.
In the morning after nocturnal GHB administration, the TRYCATs indolelactic acid, kynurenine, kynurenic acid, 3-hydroxykynurenine, and quinolinic acid; the 3-hydroxykynurenine to kynurenic acid ratio; and the CAR were significantly reduced (P < 0.05-0.001, Benjamini-Hochberg corrected). The quinolinic acid to kynurenic acid ratio was reduced by trend. Serotonin, tryptophan, and BDNF levels, as well as PANAS scores in the morning, remained unchanged after a nocturnal GHB challenge.
GHB has post-acute effects on peripheral biomarkers of neuropsychiatric disorders, which might be a model to explain some of its therapeutic effects in disorders involving neuro-immunological pathologies. This study was registered at ClinicalTrials.gov as NCT02342366.
An alternative control regimen for drug-resistant parasites is combination deworming, where two drugs with different modes of action are administered simultaneously to target the same parasite. Few ...studies have investigated this in equine cyathostomins. We previously reported that an oxibendazole (OBZ) and pyrantel pamoate (PYR) combination was not sustainable against a cyathostomin population with high levels of OBZ and PYR resistance. This study consisted of a field study and two computer simulations to evaluate the efficacy of a moxidectin-oxibendazole (MOX-OBZ) combination against the same cyathostomin population. In the field study, anthelmintic treatments occurred when ten horses exceeded 100 eggs per gram. Fecal egg counts and efficacy evaluations were performed every two weeks. The two simulations utilized weather data as well as equine and parasite population parameters from the field study. The first simulation repeated the treatment schedule used in the field study over a 40 year period. The second evaluated efficacies of combination treatments using selective therapy over 40 years. In the field study, efficacies of MOX and both combination treatments were 100%. The egg reappearance period for MOX was 16 weeks, and the two combination treatments were 12 and 18 weeks. The first (46.7%) and last (40.1%) OBZ efficacies were not significantly different from each other. In the simulation study, the combination treatment delayed MOX resistance development compared to when MOX was used as a single active. This occurred despite the low efficacy of OBZ. The second set of simulations identified combination treatments used with selective therapy to be the most effective at delaying MOX resistance. Overall, this study supports the use of combination treatment against drug-resistant cyathostomins, when one of the actives exhibits high efficacy, and demonstrates benefits of this approach despite substantially lowered efficacy of the other active ingredient.
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•Oxibendazole-moxidectin combination treatments were 100% effective.•Oxibendazole efficacies (<50%) did not differ pre and post combination treatment.•The model observed oxibendazole-moxidectincombinationto delaymoxidectin resistance.•Combination use in selective therapy delayed resistance most effectively.
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