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1.
Journal of Chemical Education ; 99(5):2079-2085, 2022.
Article in English | English Web of Science | ID: covidwho-1882728

ABSTRACT

In the past few decades, chemistry has evolved to interact withvarious disciplines to synergistically help tackle global challenges. This, in turn,requires that newer generations of chemistry students are trained to be moreflexible in accepting and coordinating new concepts. In this experiment, pineapplewas used as a key model to allow for the incorporation of multiple importantconcepts into the laboratory. Using carotenoid separation as a main goal, columnchromatography, UV-vis spectroscopy, thin-layer chromatography, high-perform-ance liquid chromatography, mass spectrometry, and principal component analysiscan be included in a cohesive laboratory experiment. On the other hand, theseactivities were also designed to be modular, thus allowing instructors to add,remove, or modify the contents in a highly customizable manner. This makes ithighly versatile and amenable to uncertain situations like unexpected universityclosure due to COVID-19 related lockdown. Overall, this laboratory experimentserves as a practical example of how chemistry can help solve real-world problems while also allowing highflexibility in teaching management

2.
Molecules ; 27(10)2022 May 21.
Article in English | MEDLINE | ID: covidwho-1875716

ABSTRACT

Nowadays, many individuals, whether healthy or diagnosed with disease, tend to expose themselves to various easily accessible natural products in hopes of benefiting their health and well-being. Mediterranean populations have traditionally used olive oil not only in nutrition but also in cosmetics, including skincare. In this study, the phenolic profile-composed of twelve compounds altogether, including the secoiridoids oleocanthal (OCAL) and oleacein (OCEIN)-of extra virgin olive oil (EVOO) from autochthonous cultivars from Croatia was determined using 1H qNMR spectroscopy and HPLC-DAD analysis, and its biological activity was investigated in melanoma cell lines. The EVOO with the highest OCEIN content had the strongest anti-cancer activity in A375 melanoma cells and the least toxic effect on the non-cancerous keratocyte cell line (HaCaT). On the other hand, pure OCAL was shown to be more effective and safer than pure OCEIN. Post-treatment with any of the EVOO phenolic extracts (EVOO-PEs) enhanced the anti-cancer effect of the anti-cancerous drug dacarbazine (DTIC) applied in pre-treatment, while they did not compromise the viability of non-cancerous cells. The metastatic melanoma A375M cell line was almost unresponsive to the EVOO-PEs themselves, as well as to pure OCEIN and OCAL. Our results demonstrate that olive oils and/or their compounds may have a potentially beneficial effect on melanoma treatment. However, their usage can be detrimental or futile, especially in healthy cells, due to inadequately applied concentrations/combinations or the presence of resistant cells.


Subject(s)
Iridoids , Melanoma , Dacarbazine , Humans , Iridoids/pharmacology , Melanoma/drug therapy , Olive Oil/chemistry , Plant Oils/chemistry , Plant Oils/pharmacology
3.
Int J Mol Sci ; 23(10)2022 May 22.
Article in English | MEDLINE | ID: covidwho-1875645

ABSTRACT

Actinidia latifolia is one of the very few kiwifruit genotypes with extremely high ascorbic acid (AsA) content. However, a transcriptome atlas of this species is lacking. The accumulation of AsA during fruit development and ripening and the associated molecular mechanisms are still poorly understood. Herein, dynamic changes in AsA content at six different stages of A. latifolia fruit development and ripening were determined. AsA content of A. latifolia fruit reached 1108.76 ± 35.26 mg 100 g-1 FW at full maturity. A high-quality, full-length (FL) transcriptome of A. latifolia was successfully constructed for the first time using third-generation sequencing technology. The transcriptome comprises 326,926 FL non-chimeric reads, 15,505 coding sequences, 2882 transcription factors, 18,797 simple sequence repeats, 3328 long noncoding RNAs, and 231 alternative splicing events. The genes involved in AsA biosynthesis and recycling pathways were identified and compared with those in different kiwifruit genotypes. The correlation between the AsA content and expression levels of key genes in AsA biosynthesis and recycling pathways was revealed. LncRNAs that participate in AsA-related gene expression regulation were also identified. Gene expression patterns in AsA biosynthesis and metabolism exhibited a trend similar to that of AsA accumulation. Overall, this study paves the way for genetic engineering to develop kiwifruits with super-high AsA content.


Subject(s)
Actinidia , Actinidia/genetics , Actinidia/metabolism , Ascorbic Acid/metabolism , Fruit/metabolism , Gene Expression Regulation, Plant , Transcriptome
4.
Foods ; 11(10):1452, 2022.
Article in English | ProQuest Central | ID: covidwho-1871540

ABSTRACT

Flavonoids are significant antioxidant and anti-inflammatory agents and have multiple potential health applications. Moringa oleifera is globally recognized for its nutritional and pharmacological properties, correlated to the high flavonoid content in its leaves. However, the bioactive compounds found in plants may vary according to the cultivation, origin, season, and extraction process used, making it difficult to extract reliable raw material. Hence, this study aimed to standardize the best cultivation and harvest season in Brazil and the best extraction process conditions to obtain a flavonoid-rich extract from M. oleifera as a final product. Firstly, ultrasound-assisted extraction (UAE) was optimized to reach the highest flavonoid content by three-level factorial planning and response surface methodology (RSM). The optimal cultivation condition was mineral soil fertilizer in the drought season, and the optimized extraction was with 80% ethanol and 13.4 min of extraction time. The flavonoid-rich extract was safe and significantly decreased reactive oxygen species (ROS) and nitric oxide (NO) in LPS-treated RAW 264.7 cells. Lastly, the major flavonoids characterized by HPLC-ESI-QTRAP-MS/MS were compounds derived from apigenin, quercetin, and kaempferol glycosides. The results confirmed that it was possible to standardize the flavonoid-rich extract leading to a standardized and reliable raw material extracted from M. oleifera leaves.

5.
Water ; 14(10):1560, 2022.
Article in English | ProQuest Central | ID: covidwho-1870966

ABSTRACT

Pharmaceuticals and personal care products (PPCPs) were investigated in five wastewater treatment plants (WWTPs), groundwater, irrigated soils, and plants in Amman and Al-Balqa governorates in Jordan. PPCPs were extracted from water samples by solid-phase extraction (SPE) and analyzed by high-performance liquid chromatography coupled with tandem mass spectrometry (HPLC–MS/MS). Carbamazepine, ciprofloxacin, ceftiofur, diclofenac, erythromycin, lincomycin, ofloxacin, pyrimthamine, spiramycin, sulfamethoxazole, sulfapyridine, testosterone, trimethoprim, and thiamphenicol were detected in all raw wastewaters in μg/L, whereas 45 PPCPs were below the detection limits (<0.02 μg/L) in all samples. Na`ur and Abu Nuseir WWTPs showed high PPCPs removal efficiencies in comparison with AL-Baqa`a, Salt, and Fuhais-Mahis WWTPs. Boqorreya spring showed signs of contamination by Salt WWTP effluents as a result of mixing. Irrigation with effluents showed higher carbamazepine concentrations in soils at the top soil layers (0 to 20 cm) in all farms than its concentrations at the root zone (20 to 40 cm) by using drip irrigation system with various plants. In plants, carbamazepine concentration was only detected in high concentration level in mint leaves. In the same farm, diclofenac concentration was detected only in olives and not in twigs and leaves, indicating a high rate of plant uptake especially during the olive’s growth period. Furthermore, plant fruits, leaves, and stems left on the farm after harvesting are generally consumed by cattle, which means entering the food chain of humans.

6.
Antioxidants (Basel) ; 11(6)2022 May 27.
Article in English | MEDLINE | ID: covidwho-1869455

ABSTRACT

The molecular constituents of Camellia sinensis, in particular epigallocatechin-3-O-gallate (EGCG) and, more remarkably, the galloylated theaflavins, mainly theaflavin-3,3'-di-O-gallate (TF-3,3'-DG), have been reported to inhibit SARS-CoV-2 3-chymotrypsin-like protease (3CLpro), an enzyme required for the cleavage of its polyproteins, to produce vital individual functional proteins for viral cell replication. Our results for total catechin content revealed the values of 174.72, 200.90, and 211.75 mg/g dry weight (DW) in spring, and the values of 183.59, 191.36, and 215.09 mg/g DW in summer, for tea plantation zones 1, 2, and 3, respectively. For the TF-3,3'-DG content, the values of 2.68, 1.13, and 3.72 mg/g DW were observed in spring, and the values of 3.78, 2.06, and 8.91 mg/g DW in summer for zones 1, 2, and 3, respectively. In the same zone, different contents of TF-3,3'-DG were observed across plucking months of April, June, and August, with values of 1.13, 2.77, and 4.18 mg/g DW, respectively, showing higher values in summer. Different values of TF-3,3'-DG contents were also observed in the same tea plantation zone but from different plant parts, revealing higher values in the bud and the first and second leaves (3.62 mg/g DW) and lower values in the third and fourth leaves (1.14 mg/g DW). The TF-3,3'-DG content increased from 3.31 to 4.98 mg/g DW with increased fermentation time from 1 to 3 h, respectively, and increased for lower temperature and longer fermentation time. The aim of this study was to investigate the processing conditions that lead to maximum TF-3,3'-DG content and, given its potential impact as an inhibitor of the 3CLpro enzyme, to create a novel antiviral Azorean black tea.

7.
Journal of Chemical Education ; 2021.
Article in English | Scopus | ID: covidwho-1829942

ABSTRACT

In the past few decades, chemistry has evolved to interact with various disciplines to synergistically help tackle global challenges. This, in turn, requires that newer generations of chemistry students are trained to be more flexible in accepting and coordinating new concepts. In this experiment, pineapple was used as a key model to allow for the incorporation of multiple important concepts into the laboratory. Using carotenoid separation as a main goal, column chromatography, UV-vis spectroscopy, thin-layer chromatography, high-performance liquid chromatography, mass spectrometry, and principal component analysis can be included in a cohesive laboratory experiment. On the other hand, these activities were also designed to be modular, thus allowing instructors to add, remove, or modify the contents in a highly customizable manner. This makes it highly versatile and amenable to uncertain situations like unexpected university closure due to COVID-19 related lockdown. Overall, this laboratory experiment serves as a practical example of how chemistry can help solve real-world problems while also allowing high flexibility in teaching management. ©

8.
Comput Struct Biotechnol J ; 20: 2082-2090, 2022.
Article in English | MEDLINE | ID: covidwho-1803848

ABSTRACT

Tumor necrosis factor (TNF) is a homotrimer that has two spatially distinct binding regions, three lectin-like domains (LLD) at the TIP of the protein and three basolaterally located receptor-binding sites, the latter of which are responsible for the inflammatory and cell death-inducing properties of the cytokine. Solnatide (a.k.a. TIP peptide, AP301) is a 17-mer cyclic peptide that mimics the LLD of human TNF which activates the amiloride-sensitive epithelial sodium channel (ENaC) and, as such, recapitulates the capacity of TNF to enhance alveolar fluid clearance, as demonstrated in numerous preclinical studies. TNF and solnatide interact with glycoproteins and these interactions are necessary for their trypanolytic and ENaC-activating activities. In view of the crucial role of ENaC in lung liquid clearance, solnatide is currently being evaluated as a novel therapeutic agent to treat pulmonary edema in patients with moderate-to-severe acute respiratory distress syndrome (ARDS), as well as severe COVID-19 patients with ARDS. To facilitate the description of the functional properties of solnatide in detail, as well as to further target-docking studies, we have analyzed its folding properties by NMR. In solution, solnatide populates a set of conformations characterized by a small hydrophobic core and two electrostatically charged poles. Using the structural information determined here and also that available for the ENaC protein, we propose a model to describe solnatide interaction with the C-terminal domain of the ENaCα subunit. This model may serve to guide future experiments to validate specific interactions with ENaCα and the design of new solnatide analogs with unexplored functionalities.

9.
Food Chem X ; 14: 100302, 2022 Jun 30.
Article in English | MEDLINE | ID: covidwho-1796842

ABSTRACT

Kombucha, originated in China 2000  years ago, is a sour and sweet-tasted drink, prepared traditionally through fermentation of black tea. During the fermentation of kombucha, consisting of mainly acidic compounds, microorganisms, and a tiny amount of alcohol, a biofilm called SCOBY forms. The bacteria in kombucha has been generally identified as Acetobacteraceae. Kombucha is a noteworthy source of B complex vitamins, polyphenols, and organic acids (mainly acetic acid). Nowadays, kombucha is tended to be prepared with some other plant species, which, therefore, lead to variations in its composition. Pre-clinical studies conducted on kombucha revealed that it has desired bioactivities such as antimicrobial, antioxidant, hepatoprotective, anti-hypercholestorelomic, anticancer, anti-inflammatory, etc. Only a few clinical studies have been also reported. In the current review, we aimed to overhaul pre-clinical bioactivities reported on kombucha as well as its brief compositional chemistry. The literature data indicate that kombucha has valuable biological effects on human health.

10.
Beverages ; 8(1):13, 2022.
Article in English | ProQuest Central | ID: covidwho-1760334

ABSTRACT

(1) Background: beverages based on extracts from Camellia sinensis are popular worldwide. Due to an increasing number of processed teas on the market, there is a need to develop unified classification standards based on chemical analysis. Meanwhile, phytochemical characterizations are mainly performed on tea samples from China (~80%). Hence, data on teas of other provenances is recommended. (2) Methods: in the present investigation, we characterized lyophilised extracts obtained by infusion, maceration and methanolic extraction derived from tea samples from China, Japan, Sri Lanka and Portugal by phytochemistry (catechins, oxyaromatic acids, flavonols, alkaloids and theanine). The real benefits of drinking the tea were analysed based on the bioavailability of the determined phytochemicals. (3) Results: the infusions revealed the highest total phenolic contents (TPC) amounts, while methanolic extracts yielded the lowest. The correlation matrix indicated that the levels of phenolic compounds were similar in the infusions and methanolic samples, while extractions made by maceration were significantly different. The differences could be partially explained by the different amounts of (-)-epigallocatechin gallate (EGCG), (-)-epicatechin gallate (ECG) and gallic acids (GA). The catechin percentages were significantly lower in the macerations, especially the quantity of EGCG decreases by 4- to 5-fold after this process. (4) Conclusions: the results highlight the importance of the processing methodology to obtain “instant tea”;the composition of the extracts obtained with the same methodology is not significantly affected by the provenance of the tea. However, attention should be drawn to the specificities of the Japanese samples (the tea analysed in the present work was of Sencha quality). In contrast, the extraction methodology significantly affects the phytochemical composition, especially concerning the content of polyphenols. As such, our results indicate that instant tea classification based on chemical composition is sensible, but there is a need for a standard extraction methodology, namely concerning the temperature and time of contact of the tea leaves with the extraction solvent.

11.
Biomed Chromatogr ; 36(7): e5370, 2022 Jul.
Article in English | MEDLINE | ID: covidwho-1748780

ABSTRACT

Ribavirin is a synthetic, broad-spectrum antiviral drug. Ribavirin is recommended as an antiviral drug in the Interim Guidance for Diagnosis and Treatment (the seventh edition) of COVID-19. The ribavirin levels in red blood cells may be closely related to both its efficacy and adverse drug reactions. In this study, a simple and fast HPLC-UV method was established to determine the concentrations of total ribavirin in the red blood cells of 13 patients with COVID-19. Phosphorylated ribavirin was dephosphorylated by phosphatase incubation to obtain the total amount of ribavirin in red blood cells. The chromatographic column was an Atlantis C18 . The recoveries were 85.45-89.05% at three levels. A good linear response was from 1 to 200 µg/ml, with a correlation coefficient of r2 = 0.9991. The concentration of total ribavirin in the red blood cells of the patients ranged from 30.83 to 133.34 µg/ml. The same samples without phosphatase incubation ranged from 4.07 to 20.84 µg/ml. About 85% of ribavirin was phosphorylated in red blood cells. In addition, we observed changes in these patients' hematological parameters and found that the erythrocyte, hemoglobin and hematocrit declined to the lowest levels on the fifth day after discontinuation of ribavirin (p < 0.05).


Subject(s)
COVID-19 , Ribavirin , Antiviral Agents , COVID-19/drug therapy , Chromatography, High Pressure Liquid/methods , Erythrocytes , Humans , Phosphoric Monoester Hydrolases/analysis , Ribavirin/analysis , Ribavirin/therapeutic use
12.
Microchem J ; 178: 107400, 2022 Jul.
Article in English | MEDLINE | ID: covidwho-1747680

ABSTRACT

Nowadays, all researchers are focused on combating the pandemic COVID-19. According to recent statistics, most patients are managed at home. An over-the-counter (OTC) triple action formula containing paracetamol (PAR), aspirin (ASP), and diphenhydramine (DIPH) is widely prescribed for pain, fever and as night-time sleep aid. For COVID-19 patients, this combination is now suggested as part of symptomatic therapy and prophylaxis. In this work, two simple liquid chromatographic approaches were designed for simultaneous determination of PAR, ASP, and DIPH in Excedrin® PM caplets, beside three specified official toxic impurities, namely, p-aminophenol, p-nitrophenol, and salicylic acid. The first method comprised high-performance thin-layer chromatographic separation coupled with densitometric quantification, on silica gel HPTLC 60 F254 aluminium sheets as the stationary phase, ethyl acetate-methanol-aqueous ammonium hydroxide (10.0: 2.0: 0.1, by volume) as the developing system and scanning was performed at 210.0 nm. The second one is a high-performance liquid chromatography coupled with diode array detector. Successful separation of the six components was performed on XTerra C18 column with isocratic elution of mobile phase 0.1% triethylamine acidified water: methanol (70:30, v/v) adjusted with o-phosphoric acid to pH 3.0 and methanol (90:10, v/v) with flow rate programming and detection at 210.0 nm. Validation of the proposed methods was performed according to ICH guidelines. Both methods were successfully used for quality control of the cited drugs in their marketed formulation. Moreover, the in-vitro release study was monitored using the proposed HPLC-DAD method. The greenness profile of the proposed methods was assessed and comparatively evaluated through various assessment tools, specifically; the analytical eco-scale system, national environmental method index (NEMI), green analytical procedure index (GAPI) and analytical greenness (AGREE) metric.

13.
Biomed Chromatogr ; 36(6): e5365, 2022 Jun.
Article in English | MEDLINE | ID: covidwho-1739127

ABSTRACT

Favipiravir is a potential antiviral medication that has been recently licensed for Covid-19 treatment. In this work, a gadolinium-based magnetic ionic liquid was prepared and used as an extractant in dispersive liquid-liquid microextraction (DLLME) of favipiravir in human plasma. The high enriching ability of DLLME allowed the determination of favipiravir in real samples using HPLC/UV with sufficient sensitivity. The effects of several variables on extraction efficiency were investigated, including type of extractant, amount of extractant, type of disperser and disperser volume. The maximum enrichment was attained using 50 mg of the Gd-magnetic ionic liquid (MIL) and 150 µl of tetrahydrofuran. The Gd-based MIL could form a supramolecular assembly in the presence of tetrahydrofuran, which enhanced the extraction efficiency of favipiravir. The developed method was validated according to US Food and Drug Administration bioanalytical method validation guidelines. The coefficient of determination was 0.9999, for a linear concentration range of 25 to 1.0 × 105  ng/ml. The percentage recovery (accuracy) varied from 99.83 to 104.2%, with RSD values (precision) ranging from 4.07 to 11.84%. The total extraction time was about 12 min and the HPLC analysis time was 5 min. The method was simple, selective and sensitive for the determination of favipiravir in real human plasma.


Subject(s)
COVID-19 , Ionic Liquids , Liquid Phase Microextraction , Amides , COVID-19/drug therapy , Chromatography, High Pressure Liquid/methods , Furans , Gadolinium , Humans , Liquid Phase Microextraction/methods , Magnetic Phenomena , Pyrazines
14.
Int J Environ Res Public Health ; 19(5)2022 02 24.
Article in English | MEDLINE | ID: covidwho-1736897

ABSTRACT

The impact of globalization on beekeeping brings new economic, scientific, ecological and social dimensions to this field The present study aimed to evaluate the chemical compositions of eight propolis extracts from Romania, and their antioxidant action and antimicrobial activity against seven species of bacteria, including pathogenic ones: Staphylococcus aureus, Bacillus cereus, Bacillus subtilis, Pseudomonas aeruginosa, Escherichia coli, Listeria monocytogenes and Salmonella enterica serovar Typhimurium. The phenolic compounds, flavonoids and antioxidant activity of propolis extracts were quantified; the presence of flavones and aromatic acids was determined. Quercetin and rutin were identified by HPLC analysis and characterized using molecular descriptors. All propolis samples exhibited antibacterial effects, especially against P. aeruginosa and L. monocytogenes. A two-way analysis of variance was used to evaluate correlations among the diameters of the inhibition zones, the bacteria used and propolis extracts used. Statistical analysis demonstrated that the diameter of the inhibition zone was influenced by the strain type, but no association between the propolis origin and the microbial activity was found.


Subject(s)
Propolis , Anti-Bacterial Agents/pharmacology , Antioxidants/chemistry , Antioxidants/pharmacology , Bacillus cereus , Escherichia coli , Microbial Sensitivity Tests , Plant Extracts/pharmacology , Propolis/pharmacology , Pseudomonas aeruginosa , Romania
15.
J Pharm Biomed Anal ; 214: 114693, 2022 May 30.
Article in English | MEDLINE | ID: covidwho-1701941

ABSTRACT

Antiviral drugs have gained much more attention in recent years due to severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection and many drug candidates are currently under investigation in order to end pandemic. Molnupiravir, a prodrug of the synthetic nucleoside derivative N4-hydroxycytidine, is one of the promising candidates for SARS-CoV-2 treatment. In this study, a RP-HPLC method was developed for the determination of Molnupiravir and applied for in vitro permeability studies of self-emulsifying drug delivery system (SEDDS) formulations using Caco-2 cell line. Discovery® HS C18 Column (75 ×4.6 mm, 3 µm) was used at 30 °C. Isocratic elution was performed with ACN:water (20:80 v/v) mixture. The flow rate was 0.5 mL/min and UV detection was at 240 nm. Molnupiravir eluted within 5 min. Molnupiravir was exposed to thermal, photolytic, hydrolytic, and oxidative stress conditions. Peak homogeneity data of Molnupiravir in the stressed samples peak obtained using photodiode array detector, in the stressed sample chromatograms, demonstrated the specificity of the method for their estimation in presence of degradants. The developed method was validated according to the International Council for Harmonisation (ICH) guidelines and found to be linear within the range 0.1-60.0 µg/mL. The method was simple, rapid, selective, sensitive, accurate, precise, robust and rugged. Thus, it was applied successfully for permeability quantitation of Molnupiravir in nanoformulations. The apparent permeability of Molnupiravir in SEDDS formulations, which have droplet size under 350 nm, was calculated as 3.20 ± 0.44 × 10-6 cm/s.


Subject(s)
COVID-19 , COVID-19/drug therapy , Caco-2 Cells , Chromatography, High Pressure Liquid/methods , Cytidine/analogs & derivatives , Drug Stability , Humans , Hydroxylamines , Permeability , Pharmaceutical Preparations , Reproducibility of Results , SARS-CoV-2
16.
J Pharm Anal ; 11(6): 709-716, 2021 Dec.
Article in English | MEDLINE | ID: covidwho-1611880

ABSTRACT

The Lianhua Qingwen (LHQW) capsule is a popular traditional Chinese medicine for the treatment of viral respiratory diseases. In particular, it has been recently prescribed to treat infections caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). However, due to its complex composition, little attention has been directed toward the analysis of chemical constituents present in the LHQW capsule. This study presents a reliable and comprehensive approach to characterizing the chemical constituents present in LHQW by high-performance liquid chromatography-Q Exactive-Orbitrap mass spectrometry (HPLC-Q Exactive-Orbitrap-MS) coupled with gas chromatography-mass spectrometry (GC-MS). An automated library alignment method with a high mass accuracy (within 5 ppm) was used for the rapid identification of compounds. A total of 104 compounds, consisting of alkaloids, flavonoids, phenols, phenolic acids, phenylpropanoids, quinones, terpenoids, and other phytochemicals, were successfully characterized. In addition, the fragmentation pathways and characteristic fragments of some representative compounds were elucidated. GC-MS analysis was conducted to characterize the volatile compounds present in LHQW. In total, 17 compounds were putatively characterized by comparing the acquired data with that from the NIST library. The major constituent was menthol, and all the other compounds were terpenoids. This is the first comprehensive report on the identification of the major chemical constituents present in the LHQW capsule by HPLC-Q Exactive-Orbitrap-MS, coupled with GC-MS, and the results of this study can be used for the quality control and standardization of LHQW capsules.

17.
EPMA J ; 12(4): 449-475, 2021 Dec.
Article in English | MEDLINE | ID: covidwho-1557745

ABSTRACT

Over the last two decades, a large number of non-communicable/chronic disorders reached an epidemic level on a global scale such as diabetes mellitus type 2, cardio-vascular disease, several types of malignancies, neurological and eye pathologies-all exerted system's enormous socio-economic burden to primary, secondary, and tertiary healthcare. The paradigm change from reactive to predictive, preventive, and personalized medicine (3PM/PPPM) has been declared as an essential transformation of the overall healthcare approach to benefit the patient and society at large. To this end, specific biomarker panels are instrumental for a cost-effective predictive approach of individualized prevention and treatments tailored to the person. The source of biomarkers is crucial for specificity and reliability of diagnostic tests and treatment targets. Furthermore, any diagnostic approach preferentially should be noninvasive to increase availability of the biomaterial, and to decrease risks of potential complications as well as concomitant costs. These requirements are clearly fulfilled by tear fluid, which represents a precious source of biomarker panels. The well-justified principle of a "sick eye in a sick body" makes comprehensive tear fluid biomarker profiling highly relevant not only for diagnostics of eye pathologies but also for prediction, prognosis, and treatment monitoring of systemic diseases. One prominent example is the Sicca syndrome linked to a cascade of severe complications that include dry eye, neurologic, and oncologic diseases. In this review, protein profiles in tear fluid are highlighted and corresponding biomarkers are exemplified for several relevant pathologies, including dry eye disease, diabetic retinopathy, cancers, and neurological disorders. Corresponding analytical approaches such as sample pre-processing, differential proteomics, electrophoretic techniques, high-performance liquid chromatography (HPLC), enzyme-linked immuno-sorbent assay (ELISA), microarrays, and mass spectrometry (MS) methodology are detailed. Consequently, we proposed the overall strategies based on the tear fluid biomarkers application for 3P medicine practice. In the context of 3P medicine, tear fluid analytical pathways are considered to predict disease development, to target preventive measures, and to create treatment algorithms tailored to individual patient profiles.

18.
J Infect Chemother ; 28(1): 73-77, 2022 Jan.
Article in English | MEDLINE | ID: covidwho-1525852

ABSTRACT

INTRODUCTION: Measurement of blood Favipiravir (FPV) levels and accumulation of data in COVID-19 patients are critical for assessing FPV efficacy and safety. We performed a retrospective study based on measurements of blood levels of FPV and related factors in COVID-19 patients admitted to our hospital. Furthermore, we also investigated the association between blood FPV levels and uric acid level alterations before and after FPV administration. METHODS: We enrolled 27 COVID-19 patients who had received FPV treatment at Hokushin General Hospital from April 1 to December 31, 2020. Age, gender, COVID-19 severity, presence of comorbidities, and laboratory data for each subject were investigated to identify factors that correlate with blood FPV levels. Uric acid levels were measured before and after FPV administration and a difference between the levels (i.e., a change of uric acid level) was evaluated. RESULTS: When a significant univariate variable was input by the stepwise method and a combination of variables that maintained statistical superiority was searched, serum ferritin was the only factor that independently affected blood FPV level. Furthermore, in the high-FPV group (20 µg/mL or more), a significant increase in uric acid levels was observed after FPV administration. The increment value was significantly larger than that in the low-FPV group (less than 20 µg/mL). CONCLUSIONS: Ferritin level was an important independent factor inversely affecting blood FPV level. Furthermore, a high blood FPV level induced the elevation of uric acid levels in COVID-19 treatment.


Subject(s)
COVID-19 , Uric Acid , Amides , Antiviral Agents/therapeutic use , COVID-19/drug therapy , Ferritins , Humans , Pyrazines , Retrospective Studies , SARS-CoV-2
19.
Materials (Basel) ; 14(21)2021 Oct 28.
Article in English | MEDLINE | ID: covidwho-1512476

ABSTRACT

Deep Eutectic Solvents (DESs) are experiencing growing interest as substitutes of polluting organic solvents for their low or absent toxicity and volatility. Moreover, they can be formed with natural bioavailable and biodegradable molecules; they are synthesized in absence of hazardous solvents. DESs are, inter alia, successfully used for the extraction/preconcentration of biofunctional molecules from complex vegetal matrices. Onion skin is a highly abundant waste material which represents a reservoir of molecules endowed with valuable biological properties such as quercetin and its glycosylated forms. An efficient extraction of these molecules from dry onion skin from "Dorata di Parma" cultivar was obtained with water dilution of acid-based DESs. Glycolic acid (with betaine 2/1 molar ratio and L-Proline 3/1 molar ratio as counterparts) and of p-toluensulphonic acid (with benzyltrimethylammonium methanesulfonate 1/1 molar ratio)-based DESs exhibited more than 3-fold higher extraction efficiency than methanol (14.79 µg/mL, 18.56 µg/mL, 14.83 µg/mL vs. 5.84 µg/mL, respectively). The extracted quercetin was also recovered efficaciously (81% of recovery) from the original extraction mixture. The proposed extraction protocol revealed to be green, efficacious and selective for the extraction of quercetin from onion skin and it could be useful for the development of other extraction procedures from other biological matrixes.

20.
Int J Mol Sci ; 22(21)2021 Oct 26.
Article in English | MEDLINE | ID: covidwho-1512374

ABSTRACT

Nucleoside kinases (NKs) are key enzymes involved in the in vivo phosphorylation of nucleoside analogues used as drugs to treat cancer or viral infections. Having different specificities, the characterization of NKs is essential for drug design and nucleotide analogue production in an in vitro enzymatic process. Therefore, a fast and reliable substrate screening method for NKs is of great importance. Here, we report on the validation of a well-known luciferase-based assay for the detection of NK activity in a 96-well plate format. The assay was semi-automated using a liquid handling robot. Good linearity was demonstrated (r² > 0.98) in the range of 0-500 µM ATP, and it was shown that alternative phosphate donors like dATP or CTP were also accepted by the luciferase. The developed high-throughput assay revealed comparable results to HPLC analysis. The assay was exemplarily used for the comparison of the substrate spectra of four NKs using 20 (8 natural, 12 modified) substrates. The screening results correlated well with literature data, and additionally, previously unknown substrates were identified for three of the NKs studied. Our results demonstrate that the developed semi-automated high-throughput assay is suitable to identify best performing NKs for a wide range of substrates.


Subject(s)
Nucleosides/metabolism , Phosphotransferases/metabolism , Adenosine Triphosphate/metabolism , Animals , Drosophila melanogaster/metabolism , Drug Evaluation, Preclinical/methods , High-Throughput Screening Assays/methods , Humans , Luciferases/metabolism , Phosphorylation/physiology , Substrate Specificity
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