Selected pharmaceutical analysis in a wastewater treatment plant during COVID-19 infection waves in South Africa

Globally, an extensive range of pharmaceuticals are consumed daily to treat a variety of illnesses and diseases. Since the occurrence of the SARS-CoV-2 virus (COVID-19) outbreak, the use of pharmaceuticals has increased drastically in order to treat and prevent infection. Studies have shown that pharmaceutical usage is largely dependent on seasonal temperatures. This was explored in the present study and was verified by the results obtained. Versatile solid phase extraction (SPE) and liquid chromatography-mass spectrometry (LC-MS) methods were developed and validated for the accurate detection of target pharmaceuticals. Method percentage recoveries ranged from 73.53-100.70%, while the limit of detection (LOD) and limit of quantification (LOQ) ranged from 0.0330-0.886 mg L−1 and 0.0990-2.68 mg L−1, respectively. Resulting concentrations of pharmaceuticals used to treat chronic ailments such as diabetes, hypertension, tuberculosis and HIV/AIDS showed consistent daily usage while pharmaceuticals used for the treatment of COVID-19 and influenza showed distinct seasonal trends. Concentrations obtained for sulfamethoxazole hydroxylamine and sulfamethoxazole ranged from 0.05215-0.3438 mg L−1 and 0.009818-0.3002 mg L−1, respectively, while concentrations quantified for prednisolone and ivermectin ranged from 0.008775-0.4482 mg L−1 and 0.008520-0.979 mg L−1, respectively. Trends also directly correlated with the total number of active COVID-19 cases experienced in South Africa during sampling periods and this was confirmed using a one-way ANOVA test. P-values obtained for sulfamethoxazole hydroxylamine, sulfamethoxazole and ivermectin were below 0.05. © 2023 The Royal Society of Chemistry.

Copper Sulfate-Induced Diphenylamine for Rapid Colorimetric Point-of-Care Detection of Contagious Pathogens Combined with Loop-Mediated Isothermal Amplification

Here, we developed a copper sulfate (CuSO4)-initiated diphenylamine (DPA)-based colorimetric strategy coupled with loop-mediated isothermal amplification (LAMP) for rapid detection of two critical contagious pathogens, SARS-CoV-2 and Enterococcus faecium. To detect the DNA, acid hydrolysis of LAMP amplicons was executed, enabling the development of a blue color. In the LAMP amplicons, the bond between the purines and deoxyribose is extremely labile. It can be broken using 70% sulfuric acid followed by phosphate group elimination, which generates a highly active keto aldehyde group. CuSO4 plays an imperative role inducing DPA to rapidly react with the keto aldehyde group, producing an intense blue color within 5 min. Moreover, low quantities such as 103 copies μL-1 of SARS-CoV-2 RNA and 102 CFU mL-1 of E. faecium were successfully detected, revealing the advantages of the introduced method. To confirm practical applicability, multiplex detection of pathogens was performed using a foldable microdevice comprising reaction and detection zones. Various reactions such as DNA extraction, LAMP, and acid hydrolysis occurred in the reaction zone. Then, colorimetric reagents (DPA, CuSO4, and ethylene glycol) contained in the detection zone were mixed with the keto aldehyde group by simply folding the microdevice, which was heated at 65 °C for 5 min for colorimetric detection. An intense blue color was developed where the target DNA was present. These results indicate that the method proposed in this study is highly suitable for point-of-care applications, especially in resource-limited settings for the rapid detection of harmful pathogens. © 2023 American Chemical Society.

Copper Sulfate-Induced Diphenylamine for Rapid Colorimetric Point-of-Care Detection of Contagious Pathogens Combined with Loop-Mediated Isothermal Amplification

Here, we developed a copper sulfate (CuSO4)-initiated diphenylamine (DPA)-based colorimetric strategy coupled with loop-mediated isothermal amplification (LAMP) for rapid detection of two critical contagious pathogens, SARS-CoV-2 and Enterococcus faecium. To detect the DNA, acid hydrolysis of LAMP amplicons was executed, enabling the development of a blue color. In the LAMP amplicons, the bond between the purines and deoxyribose is extremely labile. It can be broken using 70% sulfuric acid followed by phosphate group elimination, which generates a highly active keto aldehyde group. CuSO4 plays an imperative role inducing DPA to rapidly react with the keto aldehyde group, producing an intense blue color within 5 min. Moreover, low quantities such as 103 copies μL-1 of SARS-CoV-2 RNA and 102 CFU mL-1 of E. faecium were successfully detected, revealing the advantages of the introduced method. To confirm practical applicability, multiplex detection of pathogens was performed using a foldable microdevice comprising reaction and detection zones. Various reactions such as DNA extraction, LAMP, and acid hydrolysis occurred in the reaction zone. Then, colorimetric reagents (DPA, CuSO4, and ethylene glycol) contained in the detection zone were mixed with the keto aldehyde group by simply folding the microdevice, which was heated at 65 °C for 5 min for colorimetric detection. An intense blue color was developed where the target DNA was present. These results indicate that the method proposed in this study is highly suitable for point-of-care applications, especially in resource-limited settings for the rapid detection of harmful pathogens. © 2023 American Chemical Society

Virus removal from aqueous environments with polyelectrolyte coatings on a polypropylene fleece

The adsorption of viruses from aqueous solution is frequently performed to detect viruses. Charged filtration materials capture viruses via electrostatic interactions, but lack the specificity of biological virus-binding substances like heparin. Herein, we present three methods to immobilize heparin-mimicking, virus-binding polymers to a filter material. Two mussel-inspired approaches are used, based on dopamine or mussel-inspired dendritic polyglycerol, and post-functionalized with a block-copolymer consisting of linear polyglycerol sulfate and amino groups as anchor (lPGS-b-NH2). As third method, a polymer coating based on lPGS with benzophenone anchor groups is tested (lPGS-b-BPh). All three methods yield dense and stable coatings. A positively charged dye serves as a tool to quantitatively analyze the sulfate content on coated fleece. Especially lPGS-b-BPh is shown to be a dense polymer brush coating with about 0.1 polymer chains per nm2. Proteins adsorb to the lPGS coated materials depending on their charge, as shown for lysozyme and human serum albumin. Finally, herpes simplex virus type 1 (HSV-1) and severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) can be removed from solution upon incubation with coated fleece materials by about 90% and 45%, respectively. In summary, the presented techniques may be a useful tool to collect viruses from aqueous environments. © 2022 The Authors. Journal of Applied Polymer Science published by Wiley Periodicals LLC.

One-Pot Consecutive Reductive Amination Synthesis of Pharmaceuticals: From Biobased Glycolaldehyde to Hydroxychloroquine

Reductive amination plays a paramount role in the synthesis of amines. It is often proposed as a more ecofriendly synthesis process than the traditional SN2-type reactions of amines as it avoids toxic alkylation reagents such as alkyl halides. This work demonstrates the versatility of the reductive amination reaction via the synthesis of hydroxychloroquine (HCQ), one of the most renowned pharmaceuticals during this coronavirus pandemic. The novel green synthesis strategy is based on three consecutive reductive amination reactions conducted in a one-pot system, avoiding intermediary purification steps. Furthermore, a biobased C2 platform molecule, glycolaldehyde, was selected as a starting reagent. The newly developed reductive amination pathway was appraised using the CHEM21 Green Metric toolkit and compared with the commercially operating method. © 2022 American Chemical Society.

COVID-19 infection and neurodegeneration: Computational evidence for interactions between the SARS-CoV-2 spike protein and monoamine oxidase enzymes.

Although COVID-19 has been primarily associated with pneumonia, recent data show that its causative agent, the SARS-CoV-2 coronavirus, can infect many vital organs beyond the lungs, including the heart, kidneys and the brain. The literature agrees that COVID-19 is likely to have long-term mental health effects on infected individuals, which signifies a need to understand the role of the virus in the pathophysiology of brain disorders that is currently unknown and widely debated. Our docking and molecular dynamics simulations show that the affinity of the spike protein from the wild type (WT) and the South African B.1.351 (SA) variant towards MAO enzymes is comparable to that for its ACE2 receptor. This allows for the WT/SA⋅⋅⋅MAO complex formation, which changes MAO affinities for their neurotransmitter substrates, thereby impacting their metabolic conversion and misbalancing their levels. Knowing that this fine regulation is strongly linked with the etiology of various brain pathologies, these results are the first to highlight the possibility that the interference with the brain MAO catalytic activity is responsible for the increased neurodegenerative illnesses following a COVID-19 infection, thus placing a neurobiological link between these two conditions in the spotlight. Since the obtained insight suggests that a more contagious SA variant causes even larger disturbances, and with new and more problematic strains likely emerging in the near future, we firmly advise that the presented prospect of the SARS-CoV-2 induced neurological complications should not be ignored, but rather requires further clinical investigations to achieve an early diagnosis and timely therapeutic interventions.

Synthesis of antiviral drugs by using carbon-carbon and carbon-heteroatom bond formation under greener conditions

Antiviral medications are a branch of medicines notably used to treat that cause many significant diseases in humans and animals. This monograph mainly focuses on recent developments and synthesis of antiviral drugs using carbon-carbon and carbon-hetero bond cross-coupling chemistry. Viral infections exact several severe human diseases, accounting for remarkably high mortality rates. In this sense, academia and the pharmaceutical industry continuously search for novel compounds with better antiviral activity. The researchers face the challenge of developing greener and economical ways to synthesize these compounds and make significant progress.

Direct Imaging of Modulatory Neurotransmitters Using Synthetic Nanosensors to Understand and Treat Parkinson's Disease

Nanosensors for the optical detection of dopamine in brain slice has been validated in wildtype C57/C6 mice as well as a Parkinson's model mouse. Using nanosensors, endogenous dopamine release in striatal brain slice is triggered by electrical stimulation and quantified using microscopy. We have established the workflow for this procedure and have prepared a population of Parkinson's and wildtype mice to image in the coming quarter. We have confirmed that the method can quantitatively distinguish dopamine release between subregions of the striatum, a critical step in confirming the method for use in Parkinson's studies. COVID-19 shutdowns stalled research starting in Mar. Research facilities are reopening and the project is expected to resume this quarter.

Nutritional and industrial value of turkey meat "Meleagris Gallopavo": review

The relationship between meat consumption and health is complex and should be analyzed in detail, paying particular attention to the relevant differences that characterize the effects of different types of meat, and in several studies on poultry meat, including turkey, which is characterized by its highly digestible proteins (with low levels of collagen), and of good nutritional quality as well as unsaturated fats (found mainly in the skin and easily removed) and vitamins of group B (mainly thiamine, vitamin B6, and pantothenic acid), Minerals (such as iron, zinc, and copper) make its meat a valuable food. Through this study, it was found that there is a relationship between the consumption of turkey meat within a balanced diet and good health. Consuming it as part of a diet rich in vegetables is associated with a reduced risk of weight gain, obesity, cardiovascular disease, and type 2 diabetes. White meat (and poultry in particular) is considered moderately protective or neutral against cancer risk. The importance of poultry meat to humans has also been recognized by the Food and Agriculture Organization of the United Nations (FAO), which considers this widely available and relatively inexpensive food to be particularly beneficial in developing countries, as it can help fill in the deficiency of essential nutrients. Consumption of Turkey also contributes to the overall quality of the diet at specific ages and conditions (before conception, during pregnancy until the end of breastfeeding, during growth, and into old age) and is suitable for those with an increased need for calories and protein compared to the general population. And it was found that turkey meat contains some vital amines, which are an indicator of quality, as well as having antioxidant and antibacterial activity, and it has been proven that eating this type of meat reduces the incidence of COVID-19 disease.

Induction of Gut Microbial Tryptamine by SARS-CoV-2 in Nonhuman Primate Model Consistent with Tryptamine-Induced Model of Neurodegeneration.

The author discussed recently the possible molecular mechanisms that cause the COVID-19 disease symptoms. Here the analysis of the recent experimental data supports the hypothesis that production of the gut microbial tryptamine can be induced by the SARS-CoV-2 fecal viral activity due to the selective pressure or positive selection of tryptamine-producing microorganisms. In this report, the author suggests that the mechanism of microbial selection bases on the abilities of tryptamine to affect the viral nucleic acid. In other words, the gut microorganisms producing tryptamine are more resistant to SARS-CoV-2 fecal viral activity than microorganisms producing no tryptamine. Earlier we demonstrated the induction of neurodegeneration by tryptamine in human cells and mouse brain. Furthermore, we were able to uncover the human gut bacteria associated with Alzheimer's disease (AD) using PCR testing of human fecal samples with the new-designed primers targeting the tryptophan-tryptamine pathway. Likely, SARS-CoV-2 is one of the selective pressure factors in the cascade accelerating the neurodegenerative process in AD. This suggestion is consistent with a higher proportion of AD patients among COVID-19 related victims. Gut microbial tryptamine increase due to the viral infection-induced dysbiosis can synergize and potentiate the tryptamine cytotoxicity, necrotizing ability and other properties as a virulence factor.

Another casualty of the SARS-CoV-2 pandemic-the environmental impact.

Cemetery leachate generated by the process of cadaveric decomposition is a significant contaminant of several matrices in the cemetery environment (soil, groundwater, and surface water). The biogenic amines cadaverine and putrescine stand out among the cemetery leachate contaminants, since they are potentially carcinogenic compounds. This review article presents a discussion of possible environmental impacts caused by the increase in deaths resulting from COVID-19 as its central theme. The study also aims to demonstrate the importance of considering, in this context, some climatic factors that can alter both the time of bodily decomposition and the longevity of the virus in the environment. Additionally, some evidence for the transmission of the virus to health professionals and family members after the patient's death and environmental contamination after the burial of the bodies will also be presented. Several sources were consulted, such as scientific electronic databases (NCBI), publications by government agencies (e.g., ARPEN, Brazil) and internationally recognized health and environmental agencies (e.g., WHO, OurWorldInData.org), as well as information published on reliable websites available for free (e.g., CNN) and scientific journals related to the topic. The data from this study sounds the alarm on the fact that an increase in the number of deaths from the complications of COVID-19 has generated serious environmental problems, resulting from Cemetery leachate.

Molecular Insights into SARS-CoV2-Induced Alterations of the Gut/Brain Axis.

For a yet unknown reason, a substantial share of patients suffering from COVID-19 develop long-lasting neuropsychiatric symptoms ranging from cognitive deficits to mood disorders and/or an extreme fatigue. We previously reported that in non-neural cells, angiotensin-1 converting enzyme 2 (ACE2), the gene coding for the SARS-CoV2 host receptor, harbors tight co-expression links with dopa-decarboxylase (DDC), an enzyme involved in the metabolism of dopamine. Here, we mined and integrated data from distinct human expression atlases and found that, among a wide range of tissues and cells, enterocytes of the small intestine express the highest expression levels of ACE2, DDC and several key genes supporting the metabolism of neurotransmitters. Based on these results, we performed co-expression analyses on a recently published set of RNA-seq data obtained from SARS-CoV2-infected human intestinal organoids. We observed that in SARS-CoV2-infected enterocytes, ACE2 co-regulates not only with DDC but also with a specific group of genes involved in (i) the dopamine/trace amines metabolic pathway, (ii) the absorption of microbiota-derived L-DOPA and (iii) the absorption of neutral amino acids serving as precursors to neurotransmitters. We conclude that in patients with long COVID, a chronic infection and inflammation of small intestine enterocytes might be indirectly responsible for prolonged brain alterations.

Towards Understanding COVID-19: Molecular Insights, Co-infections, Associated Disorders, and Aging.

BACKGROUND: COVID-19 can be related to any diseases caused by microbial infection(s) because 1) co-infection with COVID-19-related virus and other microorganism(s) and 2) because metabolites produced by microorganisms such as bacteria, fungi, and protozoan can be involved in necrotizing pneumonia and other necrotizing medical conditions observed in COVID-19. OBJECTIVE: By way of illustration, the microbial metabolite of aromatic amino acid tryptophan, a biogenic amine tryptamine inducing neurodegeneration in cell and animal models, also induces necrosis. METHODS: This report includes analysis of COVID-19 positivity by zip codes in Florida and relation of the positivity to population density, possible effect of ecological and social factors on spread of COVID-19, autopsy analysis of COVID-19 cases from around the world, serum metabolomics analysis, and evaluation of autoantigenome related to COVID-19. RESULTS: In the present estimations, COVID-19 positivity percent per zip code population varied in Florida from 4.65% to 44.3% (February 2021 data). COVID-19 analysis is partially included in my book Microbial Metabolism and Disease (2021). The autoantigenome related to COVID-19 is characterized by alterations in protein biosynthesis proteins including aminoacyl-tRNA synthetases. Protein biosynthesis alteration is a feature of Alzheimer's disease. Serum metabolomics of COVID-19 positive patients show alteration in shikimate pathway metabolism, which is associated with the presence of Alzheimer's disease-associated human gut bacteria. CONCLUSION: Such alterations in microbial metabolism and protein biosynthesis can lead to toxicity and neurodegeneration as described earlier in my book Protein Biosynthesis Interference in Disease (2020).

Systemic Perturbations in Amine and Kynurenine Metabolism Associated with Acute SARS-CoV-2 Infection and Inflammatory Cytokine Responses.

We performed quantitative metabolic phenotyping of blood plasma in parallel with cytokine/chemokine analysis from participants who were either SARS-CoV-2 (+) (n = 10) or SARS-CoV-2 (-) (n = 49). SARS-CoV-2 positivity was associated with a unique metabolic phenotype and demonstrated a complex systemic response to infection, including severe perturbations in amino acid and kynurenine metabolic pathways. Nine metabolites were elevated in plasma and strongly associated with infection (quinolinic acid, glutamic acid, nicotinic acid, aspartic acid, neopterin, kynurenine, phenylalanine, 3-hydroxykynurenine, and taurine; p < 0.05), while four metabolites were lower in infection (tryptophan, histidine, indole-3-acetic acid, and citrulline; p < 0.05). This signature supports a systemic metabolic phenoconversion following infection, indicating possible neurotoxicity and neurological disruption (elevations of 3-hydroxykynurenine and quinolinic acid) and liver dysfunction (reduction in Fischer's ratio and elevation of taurine). Finally, we report correlations between the key metabolite changes observed in the disease with concentrations of proinflammatory cytokines and chemokines showing strong immunometabolic disorder in response to SARS-CoV-2 infection.

A simultaneous exploratory and quantitative amino acid and biogenic amine metabolic profiling platform for rapid disease phenotyping via UPLC-QToF-MS.

Metabolic phenotyping using mass spectrometry (MS) is being applied to ever increasing sample numbers in clinical and epidemiology studies. High-throughput and robust methods are being developed for the accurate measurement of metabolites associated with disease. Traditionally, quantitative assays have utilized triple quadrupole (QQQ) MS based methods; however, the use of such focused methods removes the ability to perform discovery-based metabolic phenotyping. An integrated workflow for the hybrid simultaneous quantification of 34 biogenic amines in combination with full scan high-resolution accurate mass (HRAM) exploratory metabolic phenotyping is presented. Primary and secondary amines are derivatized with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate prior to revered-phase liquid chromatographic separation and mass spectrometric detection. Using the HRAM-MS data, retrospective phenotypic data mining could be performed, demonstrating the versatility of HRAM-MS instrumentation in a clinical and molecular epidemiological environment. Quantitative performance was assessed using two MS detector platforms: Waters TQ-XS (QQQ; n = 3) and Bruker Impact II QToF (HRAMS-MS; n = 2) and three human biofluids (plasma, serum and urine). Finally, each platform was assessed using a certified external reference sample (NIST SRM 1950 plasma). Intra- and inter-day accuracy and precision were comparable between the QQQ and QToF instruments (<15%), with excellent linearity (R2 > 0.99) over the quantification range of 1-400 µmol L-1. Quantitative values were comparable across all instruments for human plasma, serum and urine samples, and calculated concentrations were verified against certified reference values for NIST SRM 1950 plasma as an external reference. As a real-life biological exemplar, the method was applied to plasma samples obtained from SARS-CoV-2 positive patients versus healthy controls. Both the QQQ and QToF approaches were equivalent in being able to correctly classify SARS-CoV-2 positivity. Critically, the use of HRAM full scan data was also assessed for retrospective exploratory mining of data to extract additional biogenic amines of biomarker interest beyond the 34 quantified targets.