A validated method for the analysis and profiling of ‘nyaope’ using gas chromatography – mass spectrometry

Nyaope, a Tswana word for a mixture or ‘mish-mash’, describes a drug cocktail consisting of heroin, cannabis, and on occasion other controlled substances and warfarin. It is highly addictive with extremely unpleasant side effects caused by withdrawal from the drug. It is a problem drug especially in townships in South Africa. However, its prevalence in neighbouring southern African states and further afield is not yet known. There is currently no validated method for the analysis and comparison of nyaope. We describe a validated method for the gas chromatography – mass spectrometry analysis of nyaope so that within-batch and between-batch comparisons of nyaope can successfully be made for the first time. The validated method managed an accuracy within the range 80–120%, the precision was less than 20% for all analytes and managed linearity with R2≥0.99. The detection limits for diamorphine, efavirenz, nevirapine and Δ9-tetrahydrocannabinol were 14.2, 18.6, 18.7 and 9.94 pg on column, respectively, and the limits of quantitation were 43.1, 56.3, 56.6 and 30.1 pg on column, respectively. The simulated and casework samples were successfully discriminated into original batches using the identified nyaope components, the unsupervised chemometric methods principal component analysis and hierarchical clustering, as well as chromatographic profiles.

Precision therapeutic targets for COVID-19

Beginning in late 2019, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged as a novel pathogen that causes coronavirus disease 2019 (COVID-19). SARS-CoV-2 has infected more than 111 million people worldwide and caused over 2.47 million deaths. Individuals infected with SARS-CoV-2 show symptoms of fever, cough, dyspnea, and fatigue with severe cases that can develop into pneumonia, myocarditis, acute respiratory distress syndrome, hypercoagulability, and even multi-organ failure. Current clinical management consists largely of supportive care as commonly administered treatments, including convalescent plasma, remdesivir, and high-dose glucocorticoids. These have demonstrated modest benefits in a small subset of hospitalized patients, with only dexamethasone showing demonstrable efficacy in reducing mortality and length of hospitalization. At this time, no SARS-CoV-2-specific antiviral drugs are available, although several vaccines have been approved for use in recent months. In this review, we will evaluate the efficacy of preclinical and clinical drugs that precisely target three different, essential steps of the SARS-CoV-2 replication cycle: the spike protein during entry, main protease (MPro) during proteolytic activation, and RNA-dependent RNA polymerase (RdRp) during transcription. We will assess the advantages and limitations of drugs that precisely target evolutionarily well-conserved domains, which are less likely to mutate, and therefore less likely to escape the effects of these drugs. We propose that a multi-drug cocktail targeting precise proteins, critical to the viral replication cycle, such as spike protein, MPro, and RdRp, will be the most effective strategy of inhibiting SARS-CoV-2 replication and limiting its spread in the general population.

Physical performance is enhanced in old mice fed a short term diet medicated with rapamycin, acarbose, and phenylbutyrate

Loss of physical performance, as seen in humans by decreased grip strength and overall physical fitness, is generally accepted to be a consequence of aging. Treatments to delay or reduce these changes or increase resilience to them are generally not available. In this preliminary study, 20-month-old male and female C57BL/6 mice were given either a standard mouse diet or a formulated mouse diet containing rapamycin (14 ppm), acarbose (1000 ppm), and phenylbutyrate (1000 ppm), or a diet containing one half dose of each drug, for 3 months. At the end of the study, performance on a rotarod and grip strength test was compared. In general, mice fed the full dose drug cocktail diet performed better on these assays, with significant improvements in rotarod performance in females fed the full dose cocktail and in grip strength in males fed the full dose cocktail, and females fed the low dose cocktail. These observations provide support for the concept that short term treatment with a cocktail of drugs that targets multiple aging pathways can increase resilience to aging, and suggests that this prototype cocktail could be part of a clinical therapeutic strategy for delaying age-related loss of physical performance in people. Keywords: Healthy aging, physical performance, aging processes, anti-aging drug cocktail, rapamycin, acarbose, phenylbutyrate, aging mice

Hematidrosis: Pathophysiology and Therapeutic Strategy

Background: Hematidrosis is an extremely rare and mysterious disorder. The etiology and pathophysiology of this disorder remain mostly unknown, and there is no specific therapeutic strategy available up to now.Methods: The clinical features, hemostatic and other laboratory tests, and bloody exudates of seven patients were investigated, and histologic examination of the affected skin was performed. They were treated with a new therapeutic regimen. Results: The bleeding episodes could appear under different conditions, but often following physical or emotional stress, with some prodromes. The frequency and amount of bleeding varied with each individual. The effluent bloody samples showed all the components of the peripheral blood, mingled with a few epithelial cells. Histologic examination of the affected skin showed normal sweat glands containing no blood. Each of patients was treated individually, and a multi-drug "cocktail therapy" regimen mainly consisting of beta-receptor blocker, anxiolytic, and histamine H1/H2 receptor antagonists proved successful. Although most patients could relapse within 6 to 12 months, the treatment was still effective and they were finally cured. Conclusion: nervous system-hypothalamus-autonomic nervous system pathway in association with histaminergic activation. A multi-drug "cocktail therapy" in accordance to this hypothesis would become a new effective regimen.

Evaluation of OptiFlow™-MS/MS for bioanalysis of pharmaceutical drugs and metabolites

Aim: Microflow tandem mass spectrometry-based methods have been proposed as options to improve sensitivity and selectivity while improving sample utility and solvent consumption. Here, we evaluate a newly introduced microflow source, OptiFlow™, for quantitative performance. Results/methodology: We performed a comparison of the OptiFlow and IonDrive™ sources, respectively, on the same triple quadrupole mass spectrometer. The comparison used a neat cocktail of commercially available drugs and extracted plasma samples monitoring midazolam and alprazolam metabolites. Microflow produced a 2–4× signal increase for the neat drug cocktail and a 5–10× increase for extracted plasma samples. Conclusion: The OptiFlow method consistently gave increased signal response relative to the IonDrive method and enabled a better lower limit of quantitation for defining phamacokinetics.