2. Methods
itchy, mood disorders, allergy, bipolar disorder, nasal itching, nasal congestion, allergies, depression, substance abuse
SEASONAL ALLERGIC RHINITIS, ALLERGY, RHINOCONJUNCTIVITIS, MALIGNANCIES, MULTIPLE SCLEROSIS, ALLERGIES
A double-blind, randomised, placebo-controlled clinical trial with a 2-week supplement period was carried out. Sample size calculations indicated 50 participants were required per group for the power to detect a change of 2.0 in the reflective total nasal symptom score (rTNSS) between groups (2.8 ± 3.0 vs. 4.8 ± 3.0) at the completion of the trial. The calculated effect size was 0.667; the alpha error probability was 0.05; and the power was 0.95. This study was conducted with ethics approval from the Bellberry human research ethics committee (approval number 2019-05-405) and registered with the Australian New Zealand Clinical Trials Registry (ACTRN12619001368123).This study involved two groups: an active group receiving Levagen+ and a placebo group. Levagen+ was provided by the manufacturer (Gencor Pacific, Hong Kong) and encapsulated under good manufacturing practice conditions. Each Levagen+ capsule contained 175 mg of PEA and 30 mg of excipient, and each placebo capsule contained 175 mg of maltodextrin. Both groups took two capsules daily in the morning, resulting in a total daily dose of 350 mg of PEA or maltodextrin. All trial products were enclosed in opaque capsules that were indistinguishable in appearance and administered in a consistent manner.Potential participants underwent a telehealth screening process before enrolment, which involved assessing them against specific inclusion and exclusion criteria. To be eligible for inclusion, participants needed to be 18 years of age or older, in generally good health, capable of providing informed consent, experiencing seasonal allergic rhinitis, and agreeable to maintaining their existing diet and exercise routine and refraining from using any alternative allergy relief supplements or products during the study. Female participants were required to be on a prescribed form of birth control, practicing abstinence, or post-menopausal. Additionally, all participants were required to achieve a minimum score of 3 or higher on at least 4 of the 7 baseline days for the 24 h score on the rTNSS, following the granting of consent. The rTNSS consists of 5 questions rating participants symptoms for (1) nasal congestion, (2) runny nose, (3) nasal itching, (4) sneezing, and (5) sleep difficulty over the last 12 h. Each symptom domain is scored using a four-point scale (Exclusion criteria encompassed individuals with allergies to any of the components in the active or placebo formulas, a history of chronic alcohol consumption (more than 14 alcoholic drinks per week), active smoking, substance abuse, pregnancy, or lactation. Furthermore, those with unstable or severe medical conditions, including multiple sclerosis, renal, liver, gastrointestinal, cardiovascular, diabetes, endocrine disorders, or mood disorders, such as depression and bipolar disorder, malignancies, or recent treatment for malignancies within the past two years, were excluded. Also, participants who were currently receiving or prescribed anticoagulation therapy like warfarin, heparin, dalteparin, enoxaparin, or other similar medications were ineligible for participation in the study.Eligible participants attended the study clinic for an information session before providing their written consent for inclusion in the trial. Following consent, participants underwent a health assessment that included lifestyle, current medications, and medical history. Consenting participants first recorded their symptoms twice a day for 7 days to establish rTNSS baseline scores prior to supplementation. Participants that scored 3 or more on at least 4 of the 7 baseline days for the 24 h score on the rTNSS were invited to return to the clinic and randomly assigned, via random allocation software (After receiving the product, participants were instructed to take it daily for a period of two weeks. During this time, they were also asked to complete the allergy symptom questionnaire (rTNSS) online every morning and evening. At the end of this 2-week treatment period, participants had a follow-up evaluation, which involved repeating the baseline measurements (rTNSS, RQLQ, and bloods).The primary outcome of the study was assessed by measuring the change in allergy symptoms, specifically the total rTNSS score after 12 h, which included components related to nasal congestion, sneezing, an itchy nose, and a runny nose. Secondary measurements included individual rTNSS scores, the overall score on the RQLQ (Rhinoconjunctivitis Quality of Life Questionnaire), changes in biochemical markers from baseline (cytokines, hs-CRP, FBC, histamine, and E/LFT), and the onset of action, indicating when participants first noticed an improvement in their symptoms.The analysis was conducted using either GraphPad Prism 8.0 or SPSS 25. All results were first tested for normality before any other test was conducted. Based on the distribution of the data, the appropriate statistical tests were used. In a parametric scenario, paired t-tests were used to analyse within-group differences from baseline to the endpoint. Repeated-measures ANOVA/ANCOVA were used to analyse within-group variables from all time points, with and without covariates. A general linear mixed modelling was also performed on participant data to compare group dynamics. Results were considered statistically significant if 