ABSTRACT
COVID-19 emergency required the development of specific therapeutic choices, especially for patient at higher risk to have a worst prognosis. Among the first and most widely used antivirals are remdesivir (RDV) and molnupiravir (MPV): these are both nucleoside analogs prodrugs, capable to inhibit viral RNA-dependent RNA polymerase by strand termination and a "mutational catastrophe", respectively. While RDV is only available for intravenous use in hospital, MPV is an oral drug, allowing domestic use. The circulating active metabolites of these drugs, GS-441524 and H-hydroxycitidine (NHC), respectively, are considered for the description of their pharmacokinetics (PK) and are related to their antiviral effect. Nevertheless, PK characteristics of RDV, MPV and their metabolites in the real-life use are still poorly explored, particularly due to the lack of validated methods for their quantification in human matrices. Therefore, in this work, we aimed at validating a fast, reliable and rugged ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/ MS) method for the simultaneous quantification of these prodrugs and their active metabolites in human plasma, following FDA and EMA guidelines. Sample preparation consisted in a protein precipitation protocol: 50 muL of plasma are added with 50 muL of a solution of isotope-labeled internal standards (IS) in water:methanol (50:50 v:v) and, then, with 400 muL of a mixture of acetonitrile:methanol (50:50 v:v), vortex mixed and centrifuged. After drying at 40 degreeC in vacuum centrifuge (1 h), the extracts are reconstituted with water 0.2% formic acid.Then, 5 muL of the extracts undergo UHPLC separation with a gradient run of water (Phase A) and acetonitrile:methanol 50:50 (v:v, Phase B), both with 0.2% of formic acid at 40degreeC in a core-shell reverse-phase column (Kinetex polar C18, 2.1x100 mm, 2.6 mum). The total runtime is 5 minutes and drug detection is performed by MRM, with 2 specific transitions for each compound and IS. The method fulfilled the requirements from FDA and EMA guidelines in terms of accuracy, precision, recovery, matrix effect, stability and it was applied to samples from a small cohort of patients treated with these drugs, confirming its eligibility for research and clinical use.
ABSTRACT
Smell dysfunction is one of the most frequent symptoms in COVID-19 patients. In the early stages of the disease it allows to identify positive subjects. The odorous substances recognize two different systems in the olfactory epithelium: the olfactory and the trigeminal systems that coexist and interact in the processing of sensory information. In COVID-19 patients there is an inflammatory reaction of the nasal mucosa. Infected supporting cells of the nasal mucosa release molecules that activate the local antiviral innate immune response. In fact, macrophages spread inflammatory mediators, in particular TNF-η , IL-6 and IL-1. In this study we compared IL-6 levels with the degree of olfactory disorders and with the type of unperceived odour.Materials and methodsFrom 15 March to 30 November 2020 have been selected 82 patients (45 men age 62.3 ±14.2 and 37 women age 57.1± 12.8) with only smell dysfunctions were divided into mild and moderate patients. The evaluation of the smell disorder was carried out with a 14 questionnaire relating to the perception of domestic odorous: 6 questions for olfactory sensitivity (own perfume usually sprayed, oregano, olive oil, nutella, coffee aroma, orange juice) and 8 for olfactory-trigeminal sensitivity (alcohol, fish odor, vinegar, mint (gum), toothpaste, shampoo, cheese, ammonia).The IL-6 (v.n. 0 - 7 pg/ml) was measured with chemiluminescence assay using Cobas e801 (Roche Instrumentation). Statistical analyses were performed with Wilcoxon Rank test, and Mann-Whitney test (p <0.05). ResultsThe trigeminal and olfactory sensitivity are more compromised in moderate than mild patients (p <0.05). The statistically significant differences there were in IL6 levels in moderate versus mild patients when there was an impairment of trigeminal sensitivity (p <0.05). Conclusion In this study suggested that the smell disorders in Covid-19 patients couldn't be a deficit of the olfactory central nervous pathways but could be rather than mainly associated with the inflammatory process of the nasal mucosa and that deficit of the type of domestic unperceived odour ('olfactory' or 'trigeminal' sensitivity) could indicate the degree of severity of the disease.
Subject(s)
Coronavirus Infections , Coronavirus , Pandemics , Pneumonia, Viral , Betacoronavirus , COVID-19 , Humans , SARS-CoV-2ABSTRACT
INTRODUCTION AND OBJECTIVES: We aimed to evaluate how the corona virus disease of 2019 (COVID-19) outbreak influenced emergency department (ED) admissions for urolithiasis, hospitalizations and clinical management of the hospitalized patients. PATIENTS AND METHODS: We conducted a multicentric retrospective analysis of ED admissions in three high volume urology departments (one directly involved in COVID-19 patients management and two not involved) in Rome - Italy between March and April 2020 and in the same period of 2019. Statistical analysis was conducted on the number of admissions for urolithiasis, rate of complications, hospitalization and the type of treatment received. RESULTS: 304 patients were included in the analysis. A significant reduction in the global number of patients admitted to ED for urolithiasis between 2019 and 2020 (48.8%) was noted. Moreover, regarding the choice of treatment of hospitalized patients, a statistically significant increase of stone removal procedures versus urinary drainage was reported in 2020 (p = 0.015). CONCLUSIONS: During the COVID-19 pandemic in Rome there has been a significant reduction of emergency admissions for urolithiasis. Patients admitted to ED had more complications, more frequently need hospitalization and regarding clinical management early stone removal was preferred over urinary drainage only. All the urologists should be aware that in the next months they could face an increased number of admissions for urolithiasis and manage more complicated cases.