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1.
J Agric Food Chem ; 72(19): 11278-11291, 2024 May 15.
Article in English | MEDLINE | ID: mdl-38708781

ABSTRACT

Moringa seeds are an excellent dietary source of phytochemicals (i.e., glucosinolates, GSLs; isothiocyanates, ITCs) with health-beneficial effects. Although numerous studies have been conducted on moringa seeds, the effect of germination on the regulation of GSLs remains scarcely explored. The present study investigated the dynamic changes of GSLs in moringa seeds during germination (at 25, 30, and 35 °C for 6 days in the dark) through an untargeted metabolomics approach and compared the antioxidant capacity of ungerminated and germinated moringa seeds. Our results showed that germination significantly increased the total GSL content from 150 (day 0) to 323 µmol/g (35 °C, day 6) on a dry weight (DW) basis, especially glucomoringin (GMG), the unique glucosinolate in moringa seeds, which was significantly upregulated from 61 (day 0) to 149 µmol/g DW (35 °C, day 4). The upregulation of GMG corresponded to the metabolism of tyrosine, which might be the initial precursor for the formation of GMG. In addition, germination enhanced the total ITC content from 85 (day 0) to 239 µmol SE/g DW (35 °C, day 6), indicating that germination may have also increased the activity of myrosinase. Furthermore, germination remarkably increased the total phenolic content (109-507 mg GAE/100 g DW) and antioxidant capacity of moringa seeds. Our findings suggest that moringa sprouts could be promoted as a novel food and/or ingredient rich in GMG.


Subject(s)
Germination , Glucosinolates , Moringa , Seeds , Tyrosine , Seeds/chemistry , Seeds/metabolism , Seeds/growth & development , Tyrosine/metabolism , Tyrosine/analysis , Moringa/chemistry , Moringa/metabolism , Moringa/growth & development , Glucosinolates/metabolism , Glucosinolates/analysis , Glucosinolates/chemistry , Antioxidants/metabolism , Antioxidants/chemistry , Antioxidants/analysis
2.
Int J Food Microbiol ; 415: 110631, 2024 Apr 16.
Article in English | MEDLINE | ID: mdl-38402671

ABSTRACT

Hanseniaspora vineae exhibits extraordinary positive oenological characteristics contributing to the aroma and texture of wines, especially by its ability to produce great concentrations of benzenoid and phenylpropanoid compounds compared with conventional Saccharomyces yeasts. Consequently, in practice, sequential inoculation of H. vineae and Saccharomyces cerevisiae allows to improve the aromatic quality of wines. In this work, we evaluated the impact on wine aroma produced by increasing the concentration of phenylalanine, the main amino acid precursor of phenylpropanoids and benzenoids. Fermentations were carried out using a Chardonnay grape juice containing 150 mg N/L yeast assimilable nitrogen. Fermentations were performed adding 60 mg/L of phenylalanine without any supplementary addition to the juice. Musts were inoculated sequentially using three different H. vineae strains isolated from Uruguayan vineyards and, after 96 h, S. cerevisiae was inoculated to complete the process. At the end of the fermentation, wine aromas were analysed by both gas chromatography-mass spectrometry and sensory evaluation through a panel of experts. Aromas derived from aromatic amino acids were differentially produced depending on the treatments. Sensory analysis revealed more floral character and greater aromatic complexity when compared with control fermentations without phenylalanine added. Moreover, fermentations performed in synthetic must with pure H. vineae revealed that even tyrosine can be used in absence of phenylalanine, and phenylalanine is not used by this yeast for the synthesis of tyrosine derivatives.


Subject(s)
Hanseniaspora , Wine , Wine/analysis , Fermentation , Saccharomyces cerevisiae/metabolism , Odorants/analysis , Phenylalanine/analysis , Phenylalanine/metabolism , Hanseniaspora/metabolism , Tyrosine/analysis , Tyrosine/metabolism
3.
Analyst ; 148(13): 2965-2974, 2023 Jun 26.
Article in English | MEDLINE | ID: mdl-37265393

ABSTRACT

A wearable sweat sensor, which could continuously monitor biomolecules related to the human physiological state, is emerging as a promising piece of health surveillance equipment. However, current sensors cannot simultaneously achieve a detection performance that equates to that of traditional sensors and satisfactory mechanical strength. Herein, a wearable sweat sensor with excellent detection performance and mechanical stability is designed and fabricated. Based on the integration of laser-induced graphene electrodes and a screen printing technique, this wearable sweat sensor could realize both the separate and simultaneous detection of uric acid (UA), tyrosine (Tyr), and ascorbic acid (AA) with high sensitivity. Good UA sensing performance in artificial sweat could be maintained even after 20 000 bends. In addition, the sensor can operate well in the wearing state or in a complex bovine whole blood sample. For the detection of human sweat, the changes in UA concentration after a purine-rich meal are continuously monitored and the results are in accordance with the corresponding serum UA detection results tested with a commercial serum UA meter. These results suggest its application potential in health monitoring for both gout patients and healthy humans.


Subject(s)
Sweat , Animals , Cattle , Sweat/chemistry , Uric Acid/analysis , Tyrosine/analysis , Ascorbic Acid/analysis , Humans , Wearable Electronic Devices
4.
J Sci Food Agric ; 103(2): 750-763, 2023 Jan 30.
Article in English | MEDLINE | ID: mdl-36054758

ABSTRACT

BACKGROUND: N-Carbamoyl-aspartic acid (NCA) is a critical precursor for de novo biosynthesis of pyrimidine nucleotides. To investigate the cumulative effects of maternal supplementation with NCA on the productive performance, serum metabolites and intestinal microbiota of sows, 40 pregnant sows (∼day 80) were assigned into two groups: (1) the control (CON) and (2) treatment (NCA, 50 g t-1 NCA). RESULTS: Results showed that piglets from the NCA group had heavier birth weight than those in the CON group (P < 0.05). In addition, maternal supplementation with NCA decreased the backfat loss of sows during lactation (P < 0.05). Furthermore,16S-rRNA sequencing results revealed that maternal NCA supplementation decreased the abundance of Cellulosilyticum, Fournierella, Anaerovibrio, and Oribacterium genera of sows during late pregnancy (P < 0.05). Similarly, on the 14th day of lactation, maternal supplementation with NCA reduced the diversity of fecal microbes of sows as evidenced by significantly lower observed species, Chao1, and Ace indexes, and decreased the abundance of Lachnospire, Faecalibacterium, and Anaerovorax genera, while enriched the abundance of Catenisphaera (P < 0.05). Untargeted metabolomics showed that a total of 48 differentially abundant biomarkers were identified, which were mainly involved in metabolic pathways of arginine/proline metabolism, phenylalanine/tyrosine metabolism, and fatty acid biosynthesis, etc. CONCLUSION: Overall, the results indicated that NCA supplementation regulated intestinal microbial composition of sows and serum differential metabolites related to arginine, proline, phenylalanine, tyrosine, and fatty acids metabolism that may contribute to regulating the backfat loss of sows, and the birth weight and diarrhea rate of piglets. © 2022 Society of Chemical Industry.


Subject(s)
Gastrointestinal Microbiome , Swine , Animals , Pregnancy , Female , Animal Feed/analysis , Colostrum/chemistry , Aspartic Acid/analysis , Aspartic Acid/metabolism , Aspartic Acid/pharmacology , Dietary Supplements/analysis , Birth Weight , Diet/veterinary , Lactation , Arginine/analysis , Phenylalanine/analysis , Tyrosine/analysis , Proline/analysis
5.
Anal Chim Acta ; 1232: 340447, 2022 Nov 01.
Article in English | MEDLINE | ID: mdl-36257734

ABSTRACT

Multiplexed detection of biomolecules is of great value in various fields, from disease diagnosis to food safety and environmental monitoring. However, accurate and multiplexed analyte detection is challenging to achieve in mixtures using a single device/material. In this paper, we demonstrate a machine learning (ML)-powered multimodal analytical device based on a single sensing material made of electrodeposited molybdenum polysulfide (eMoSx) on laser induced graphene (LIG) for multiplexed detection of tyrosine (TYR) and uric acid (UA) in sweat and saliva. Electrodeposition of MoSx shows an increased electrochemically active surface area (ECSA) and heterogeneous electron transfer rate constant, k0. Features are extracted from the electrochemical data in order to train ML models to predict the analyte concentration in the sample (both singly spiked and mixed samples). Different ML architectures are explored to optimize the sensing performance. The optimized ML-based multimodal analytical system offers a limit of detection (LOD) that is two orders of magnitude better than conventional approaches which rely on single peak analysis. A flexible and wearable sensor patch is also fabricated and validated on-body, achieving detection of UA and TYR in sweat over a wide concentration range. While the performance of the developed approach is demonstrated for detecting TYR and UA using eMoSx-LIG sensors, it is a general analytical methodology and can be extended to a variety of electrochemical sensors to enable accurate, reliable, and multiplexed sensing.


Subject(s)
Biosensing Techniques , Graphite , Smart Materials , Sweat/chemistry , Graphite/chemistry , Uric Acid/analysis , Saliva/chemistry , Tyrosine/analysis , Molybdenum , Lasers , Machine Learning , Biosensing Techniques/methods , Electrochemical Techniques/methods
6.
Food Chem Toxicol ; 169: 113421, 2022 Nov.
Article in English | MEDLINE | ID: mdl-36100043

ABSTRACT

Anticancer agents can cause various side effects, including tissue damages/inflammatory reactions. Drug-responsive biomarkers are essential for evaluating drug toxicity in disease processes. S100 calcium-binding proteins A8/A9 (S100A8/A9) are highly expressed in neutrophils and monocytes/macrophages accumulated at inflammatory sites and are known to be related to tissue damage/inflammation; however, their response to drug toxicity has not been reported. Herein, we investigated the effects of anticancer agents (doxorubicin, cisplatin, and docetaxel) on S100A8/A9 gene expression profiles in four representative tissues (heart, kidney, liver, and lung) in normal C57BL/6J mice. Both S100A8/A9 expression was transiently or time-dependently elevated in four tissues within 48 h after dosing of the three anticancer agents under toxicity-inducing conditions. S100A8/A9 patterns differed among agents and tissues. This result suggests that S100A8/A9 is useful for evaluating anticancer agent-induced tissue damage. Metabolomic analysis revealed that some metabolites showed temporal patterns similar to that of S100A8/A9 expression. The amounts of fumarate (doxorubicin-treated heart), tyrosine (cisplatin-treated kidney), acetylcarnosine (doxorubicin-treated liver), and 2-phosphoglycerate (docetaxel-treated lung) showed similar patterns to that of S100A8/A9 expression. Although these metabolites showed different behaviors between tissues and serum, they may be useful marker candidates for evaluating anticancer agent-induced tissue damage at an earlier stage after dosing.


Subject(s)
Antineoplastic Agents , Biomarkers, Pharmacological , Calgranulin A , Calgranulin B , Inflammation , Animals , Mice , Antineoplastic Agents/administration & dosage , Antineoplastic Agents/toxicity , Biomarkers, Pharmacological/metabolism , Calgranulin A/genetics , Calgranulin A/metabolism , Calgranulin B/genetics , Calgranulin B/metabolism , Cisplatin/administration & dosage , Cisplatin/toxicity , Docetaxel/administration & dosage , Docetaxel/toxicity , Doxorubicin/administration & dosage , Doxorubicin/toxicity , Fumarates/analysis , Inflammation/chemically induced , Inflammation/metabolism , Mice, Inbred C57BL , Tyrosine/analysis
7.
Sci Total Environ ; 850: 158019, 2022 Dec 01.
Article in English | MEDLINE | ID: mdl-35973547

ABSTRACT

Forest environment has many health benefits, and negative air ions (NAI) is one of the major forest environmental factors. Many studies have explored the effect of forest environment on cardiac autonomic nervous function, while forest NAI in the among function and the underlying mechanism still remain unclear. To explore the associations and molecular linkages between short-term exposure to forest NAI and heart rate variability (HRV), a repeated-measure panel study was conducted among 31 healthy adults. Participants were randomly selected to stay in a forest park for 3 days and 2 nights. Individual exposures including NAI were monitored simultaneously and HRV indices were measured repeatedly at the follow-up period. Urine samples were collected for non-targeted metabolomics analysis. Mixed-effect models were adopted to evaluate associations among NAI, HRV indices and metabolites. The median of NAI concentration was 68.11 (138.20) cm-3 during the study period. Short-term exposure to forest NAI was associated with the ameliorative HRV indices, especially the excitatory parasympathetic nerve. For instance, per interquartile range increase of 5-min moving average of NAI was associated with 9.99 % (95%CI: 8.95 %, 11.03 %) increase of power in high frequency. Eight metabolites were associated with NAI exposure. The down-regulated tyrosine metabolism was firstly observed, followed by other amino acid metabolic alterations. The NAI-related metabolic changes reflect the reduction of inflammation and oxidative stress. HRV indices were associated with 25 metabolites, mainly including arginine, proline and histidine metabolism. Short-term exposure to forest NAI is beneficial to HRV, especially to the parasympathetic nerve activity, by successively disturbing different metabolic pathways which mainly reflect the increased anti-inflammation and the reduced inflammation. The results will provide epidemiological evidences for developing forest therapy and improving cardiac autonomic nervous function.


Subject(s)
Air Pollutants , Particulate Matter , Adult , Air Pollutants/analysis , Arginine/analysis , Forests , Heart Rate , Histidine/analysis , Histidine/pharmacology , Humans , Ions/analysis , Particulate Matter/analysis , Proline/analysis , Proline/pharmacology , Tyrosine/analysis , Tyrosine/pharmacology
8.
Int J Mol Sci ; 23(3)2022 Jan 22.
Article in English | MEDLINE | ID: mdl-35163145

ABSTRACT

Recently, the studies on developing sensors and biosensors-with an obvious interdisciplinary character-have drawn the attention of many researchers specializing in various fundamental, but also complex domains such as chemistry, biochemistry, physics, biophysics, biology, bio-pharma-medicine, and bioengineering. Along these lines, the present paper is structured into three parts, and is aimed at synthesizing the most relevant studies on the construction and functioning of versatile devices, of electrochemical sensors and biosensors, respectively. The first part presents examples of the most representative scientific research focusing on the role and the importance of the phenylalanine, tyrosine, and tryptophan amino acids, selected depending on their chemical structure and their impact on the central nervous system. The second part is dedicated to presenting and exemplifying conductor polymers and molecularly imprinted polymers used as sensitive materials in achieving electrochemical sensors and biosensors. The last part of the review analyzes the sensors and biosensors developed so far to detect amino acids with the aid of conductor polymers and molecularly imprinted polymers from the point of view of the performances obtained, with emphasis on the detection methods, on the electrochemical reactions that take place upon detection, and on the electroanalytical performances. The present study was carried out with a view to highlighting, for the benefit of specialists in medicine and pharmacy, the possibility of achieving and purchasing efficient devices that might be used in the quality control of medicines, as well as in studying and monitoring diseases associated with these amino acids.


Subject(s)
Biosensing Techniques/instrumentation , Electrochemical Techniques/methods , Molecular Imprinting/methods , Molecularly Imprinted Polymers/chemistry , Phenylalanine/analysis , Tryptophan/analysis , Tyrosine/analysis , Amino Acids/analysis , Polymers/chemistry
9.
Molecules ; 27(3)2022 Feb 08.
Article in English | MEDLINE | ID: mdl-35164404

ABSTRACT

Studies carried out using three different in vitro assays and a biological setting (Escherichia coil) demonstrated the antioxidant activity of Scutellaria lateriflora microshoot extract. Moreover, the extract exhibited no toxicity in a brine shrimp lethality bioassay. These results indicated that microshoots are a rich, safe source of antioxidants, which encouraged us to enhance their production in vitro. In agar and agitated cultures, two biotechnological strategies were applied: feeding the cultures with the biogenetic precursors of the phenolics-phenylalanine and tyrosine, and eliciting them with methyl jasmonate. Specific Scutellaria flavonoids and verbascoside were analysed by HPLC. Feeding with precursors (1 g/L) in agar cultures decreased the production of the metabolites. In agitated cultures, different concentrations of precursors (1.0-2.5 g/L) and the elicitor (10; 50; 100 µM) were tested. Additionally, parallel feeding with the precursor and elicitor in a concentration of 50 µM were applied. The best strategy for total flavonoid and verbascoside production was phenylalanine feeding (1.5 g/L), max. 3765 and 475 mg/100 g DW, respectively, after 7 days. This is the first report documenting the high antioxidant production in S. lateriflora microshoots after feeding with phenylalanine. Moreover, for the first time, bioreactor cultures were successfully maintained, obtaining attractive results (max. total flavonoid content 2348 and verbascoside 485 mg/100 g DW).


Subject(s)
Antioxidants/metabolism , Biotechnology , Phytochemicals/metabolism , Plant Shoots/metabolism , Scutellaria/metabolism , Acetates/analysis , Acetates/metabolism , Antioxidants/analysis , Cell Culture Techniques , Cyclopentanes/analysis , Cyclopentanes/metabolism , Flavonoids/analysis , Flavonoids/metabolism , Oxylipins/analysis , Oxylipins/metabolism , Phytochemicals/analysis , Plant Shoots/chemistry , Scutellaria/chemistry , Tyrosine/analysis , Tyrosine/metabolism
10.
J Proteome Res ; 21(3): 808-821, 2022 03 04.
Article in English | MEDLINE | ID: mdl-34365791

ABSTRACT

Hyperlipidemia is one kind of metabolic syndrome for which the treatment commonly includes simvastatin (SV). Individuals vary widely in statin responses, and growing evidence implicates gut microbiome involvement in this variability. However, the associated molecular mechanisms between metabolic improvement and microbiota composition following SV treatment are still not fully understood. In this study, combinatory approaches using ultrahigh-performance liquid chromatography coupled with hybrid triple quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF MS/MS)-based metabolomic profiling, PCR-denaturing gradient gel electrophoresis (PCR-DGGE), quantitative PCR (qPCR), and 16S rRNA gene sequencing-based gut microbiota profiling were performed to investigate the interplay of endogenous metabolites and the gut microbiota related to SV treatment. A total of 6 key differential endogenous metabolites were identified that affect the metabolism of amino acids (phenylalanine and tyrosine), unsaturated fatty acids (linoleic acid and 9-hydroxyoctadecadienoic acid (9-HODE)), and the functions of gut microbial metabolism. Moreover, a total of 22 differentially abundant taxa were obtained following SV treatment. Three bacterial taxa were identified to be involved in SV treatment, namely, Bacteroidaceae, Prevotellaceae, and Porphyromonadaceae. These findings suggested that the phenylalanine and tyrosine-associated amino acid metabolism pathways, as well as the linoleic acid and 9-HODE-associated unsaturated fatty acid metabolism pathways, which are involved in gut flora interactions, might be potential therapeutic targets for improvement in SV hypolipidemic efficacy. The mass spectrometric data have been deposited to MassIVE (https://massive.ucsd.edu/ProteoSAFe/static/massive.jsp). Username: MSV000087842_reviewer. Password: hardworkingzsr.


Subject(s)
Simvastatin , Tandem Mass Spectrometry , Animals , Bacteria/genetics , Bacteria/metabolism , Diet , Feces/microbiology , Linoleic Acids , Metabolomics/methods , Phenylalanine/analysis , RNA, Ribosomal, 16S/analysis , RNA, Ribosomal, 16S/genetics , Rats , Simvastatin/pharmacology , Tyrosine/analysis
11.
Life Sci ; 290: 120246, 2022 Feb 01.
Article in English | MEDLINE | ID: mdl-34953892

ABSTRACT

AIMS: To primarily explore the mechanism of captopril in oxidative stress and investigate the link between captopril alleviated oxidative damage and diabetic retinopathy (DR). MAIN METHODS: Human retinal microvascular endothelial cells (HRMECs) were used for in vitro experiments and cultured in a 5.5 mM or 30 mM glucose medium. Sprague-Dawley rats were used for in vivo experiments, and parts of the rats were established for diabetic groups by injected streptozotocin (n = 10, each group). Both experiments had a captopril-treated group. The levels of total cholesterol (TC), reactive oxygen species (ROS), nitric oxide (NO), and human 3-nitrotyrosine (3-NT) were detected in assay kits and ELISA. Western blotting was used to detect the expression of steroid regulatory element binding protein 2 (SREBP2), inducible nitric oxide synthase (iNOS), vascular endothelial growth factor (VEGF), and endothelial nitric oxide synthase (eNOS). Hematoxylin-eosin staining and Evans blue were used to describe retinal histopathology. KEY FINDINGS: The levels of TC, ROS, NO, and 3-NT were increased in the higher glucose groups compared with the normal controls during in vivo and in vitro experiments. Western blotting showed a higher level of SREBP2, iNOS, and VEGF and a lower eNOS level in the higher glucose groups. These results were reversed by captopril. Captopril relieved diabetic retinal vascular leakage. SIGNIFICANCE: Our study suggested that captopril alleviates oxidative damage in DR due to creating lower peroxynitrite by decreasing ROS and NO, which may provide a visible direction for DR research.


Subject(s)
Captopril/pharmacology , Diabetic Retinopathy/drug therapy , Oxidative Stress/drug effects , Animals , Captopril/metabolism , Cholesterol/analysis , Diabetes Mellitus, Experimental/drug therapy , Diabetic Retinopathy/metabolism , Disease Models, Animal , Endothelial Cells/metabolism , Glucose/metabolism , Humans , Male , Nitric Oxide/analysis , Nitric Oxide Synthase Type II/metabolism , Nitric Oxide Synthase Type III/metabolism , Oxidative Stress/physiology , Rats , Rats, Sprague-Dawley , Reactive Oxygen Species/analysis , Retina/pathology , Retinal Vessels/metabolism , Streptozocin/pharmacology , Tyrosine/analogs & derivatives , Tyrosine/analysis , Vascular Endothelial Growth Factor A/metabolism
12.
Nanotoxicology ; 16(9-10): 913-934, 2022.
Article in English | MEDLINE | ID: mdl-36774544

ABSTRACT

Inhalation of nanoparticles emitted from toner-based printing equipment (TPE), such as laser printers and photocopiers, also known as PEPs, has been associated with systemic inflammation, hypertension, cardiovascular disease, respiratory disorders, and genotoxicity. Global serum metabolomics analysis in 19 healthy TPE operators found 52 dysregulated biomolecules involved in upregulation of inflammation, immune, and antioxidant responses and downregulation of cellular energetics and cell proliferation. Here, we build on the metabolomics study by investigating the association of a panel of nine urinary OS biomarkers reflecting DNA/RNA damage (8OHdG, 8OHG, and 5OHMeU), protein/amino acid oxidation (o-tyrosine, 3-chlorotyrosine, and 3-nitrotyrosine), and lipid oxidation (8-isoprostane, 4-hydroxy nonenal, and malondialdehyde [MDA]), as well as plasma total MDA and total protein carbonyl (TPC), with several nanoparticle exposure metrics in the same 19 healthy TPE operators. Plasma total MDA, urinary 5OHMeU, 3-chlorotyrosine, and 3-nitrotyrosine were positively, whereas o-tyrosine inversely and statistically significantly associated with PEPs exposure in multivariate models, after adjusting for age and urinary creatinine. Urinary 8OHdG, 8OHG, 5OHMeU, and total MDA in urine and plasma had group mean values higher than expected in healthy controls without PEPs exposure and comparable to those of workers experiencing low to moderate levels of oxidative stress (OS). The highest exposure group had OS biomarker values, most notably 8OHdG, 8OHG, and total MDA, that compared to workers exposed to welding fumes and titanium dioxide. Particle number concentration was the most sensitive and robust exposure metric. A combination of nanoparticle number concentration and OS potential of fresh aerosols is recommended for larger scale future studies.


Subject(s)
Air Pollutants , Nanoparticles , Humans , Air Pollutants/toxicity , 8-Hydroxy-2'-Deoxyguanosine/analysis , Singapore , Nanoparticles/toxicity , Oxidative Stress , Biomarkers/analysis , Tyrosine/analysis , Inflammation , Printing, Three-Dimensional
13.
Anal Bioanal Chem ; 413(27): 6857-6866, 2021 Nov.
Article in English | MEDLINE | ID: mdl-34491394

ABSTRACT

Fast identification of pathogenic bacteria is an essential need for patient's diagnostic in hospitals and environmental monitoring of water and air quality. Bacterial cells consist of a very high amount of biological molecules whose content changes in response to different environmental conditions. The similarity between the molecular compositions of different bacterial cells limits the possibility to find unique markers to enable differentiation among species. Although many biological molecules in the cells absorb at the UV-Vis region, only a few of them can be detected in whole cells by their intrinsic fluorescence. Among these molecules are the amino acids phenylalanine, tyrosine, and tryptophan. In this work, we develop a rapid method for bacterial identification by synchronous fluorescence. We show that we can quantify the concentration for the 3 amino acids without any significant interference from other fluorophores in the cells and that we can differentiate among 6 pathogenic bacterial species by using the concentrations of their amino acids as a bacterial fingerprint. Fluorescent amino acids exist in all living cells. Therefore, this method has the potential to be applicative for the rapid identification of cells from all kinds of organisms.


Subject(s)
Amino Acids/analysis , Bacteria/chemistry , Bacteria/classification , Bacterial Typing Techniques/methods , Amino Acids/chemistry , Bacteria/isolation & purification , Bacteria/pathogenicity , Calibration , Escherichia coli/chemistry , Escherichia coli/classification , Fluorescence , Phenylalanine/analysis , Phenylalanine/chemistry , Spectrometry, Fluorescence/methods , Tryptophan/analysis , Tryptophan/chemistry , Tyrosine/analysis , Tyrosine/chemistry
14.
Biosensors (Basel) ; 11(8)2021 Aug 23.
Article in English | MEDLINE | ID: mdl-34436092

ABSTRACT

Tyrosinase (TYR, E.C. 1.14.18.1), a critical enzyme participating in melanogenesis, catalyzes the first two steps in melanin biosynthesis including the ortho-hydroxylation of L-tyrosine and the oxidation of L-DOPA. Previous pharmacological investigations have revealed that an abnormal level of TYR is tightly associated with various dermatoses, including albinism, age spots, and malignant melanoma. TYR inhibitors can partially block the formation of pigment, which are always used for improving skin tone and treating dermatoses. The practical and reliable assays for monitoring TYR activity levels are very useful for both disease diagnosis and drug discovery. This review comprehensively summarizes structural and enzymatic characteristics, catalytic mechanism and substrate preference of TYR, as well as the recent advances in biochemical assays for sensing TYR activity and their biomedical applications. The design strategies of various TYR substrates, alongside with several lists of all reported biochemical assays for sensing TYR including analytical conditions and kinetic parameters, are presented for the first time. Additionally, the biomedical applications and future perspectives of these optical assays are also highlighted. The information and knowledge presented in this review offer a group of practical and reliable assays and imaging tools for sensing TYR activities in complex biological systems, which strongly facilitates high-throughput screening TYR inhibitors and further investigations on the relevance of TYR to human diseases.


Subject(s)
Biosensing Techniques , Tyrosine/analysis , Humans , Kinetics , Melanoma , Monophenol Monooxygenase , Oxidation-Reduction , Skin Neoplasms , Spectrophotometry , Melanoma, Cutaneous Malignant
15.
Int J Mol Sci ; 22(14)2021 Jul 14.
Article in English | MEDLINE | ID: mdl-34299146

ABSTRACT

Good health, of vital importance in order to carry out our daily routine, consists of both physical and mental health. Tyrosine (Tyr) deficiency as well as its excess are issues that can affect mental health and can generate disorders such as depression, anxiety, or stress. Tyr is the amino acid (AA) responsible for maintaining good mental health, and for this reason, the present research presents the development of new electrochemical sensors modified with polypyrrole (PPy) doped with different doping agents such as potassium hexacyanoferrate (II) (FeCN), sodium nitroprusside (NP), and sodium dodecyl sulfate (SDS) for a selective and sensitive detection of Tyr. The development of the sensors was carried out by chronoamperometry (CA) and the electrochemical characterization was carried out by cyclic voltammetry (CV). The detection limits (LOD) obtained with each modified sensor were 8.2 × 10-8 M in the case of PPy /FeCN-SPCE, 4.3 × 10-7 M in the case of PPy/NP-SPCE, and of 3.51 × 10-7 M in the case of PPy/SDS-SPCE, thus demonstrating a good sensitivity of these sensors detecting L-Tyr. The validation of sensors was carried out through quantification of L-Tyr from three pharmaceutical products by the standard addition method with recoveries in the range 99.92-103.97%. Thus, the sensors present adequate selectivity and can be used in the pharmaceutical and medical fields.


Subject(s)
Carbon/chemistry , Electrodes , Pharmaceutical Preparations/analysis , Pharmaceutical Preparations/chemistry , Polymers/chemistry , Pyrroles/chemistry , Tyrosine/analysis , Electrochemical Techniques
16.
Sci Rep ; 11(1): 15036, 2021 07 22.
Article in English | MEDLINE | ID: mdl-34294750

ABSTRACT

Renal injury caused by renal ischemia and reperfusion strongly influences heart morphology, electrophysiology, and redox unbalance. The so-called cardiorenal syndrome is an important class of dysfunction since heart and kidneys are responsible for hemodynamic stability and organ perfusion through a complex network. In the present work we investigate the vibrational spectral features probed by Fourier-Transform Raman (FT-Raman) spectroscopy due to physiological alterations induced by renal ischemic reperfusion aiming to detect molecular markers related to progression of acute to chronic kidney injury and mortality predictors as well. C57BL/6J mice were subjected to unilateral occlusion of the renal pedicle for 60 min and reperfusion for 5, 8, and 15 days. Biopsies of heart and kidney tissues were analyzed. Our findings indicated that cysteine/cystine, fatty acids, methyl groups of Collagen, α-form of proteins, Tyrosine, and Tryptophan were modulated during renal ischemia and reperfusion process. These changes are consistent with fibroblast growth factors and Collagen III contents changes. Interestingly, Tyrosine and Tryptophan, precursor molecules for the formation of uremic toxins such as indoxyl sulfate and p-cresyl sulfate were also modulated. They are markers of kidney injury and their increase is strongly correlated to cardiovascular mortality. Regarding this aspect, we notice that monitoring the Tyrosine and Tryptophan bands at 1558, 1616, and 1625 cm-1 is a viable and and advantageous way to predict fatality in cardiovascular diseases both "in vivo" or "in vitro", using the real-time, multiplexing, and minimally invasive advantages of FT-Raman spectroscopy.


Subject(s)
Biomarkers , Kidney Diseases/etiology , Kidney Diseases/metabolism , Tryptophan/metabolism , Tyrosine/metabolism , Animals , Cardio-Renal Syndrome/diagnosis , Cardio-Renal Syndrome/etiology , Cardio-Renal Syndrome/metabolism , Disease Management , Disease Susceptibility , Kidney Diseases/diagnosis , Male , Mice , Organ Specificity , Reperfusion Injury/complications , Spectrum Analysis/methods , Tryptophan/analysis , Tyrosine/analysis
17.
Methods Mol Biol ; 2276: 383-396, 2021.
Article in English | MEDLINE | ID: mdl-34060056

ABSTRACT

Nitrotyrosine formation is caused by presence of reactive oxygen and nitrogen species. Nitration is a very selective process leading to specific modification of only a few tyrosines in protein molecule. 2D electrophoresis and western blotting techniques coupled with mass spectrometry are common methods used in analysis of proteome. Here we describe protocol for analysis of peroxynitrite-induced protein nitration in isolated mitochondria. Mitochondrial proteins are separated by 2D electrophoresis and transferred to nitrocellulose membrane. Membranes are then incubated with antibodies against nitrotyrosine. Positive spots are compared with corresponding Coomassie-stained gels, and protein nitration is confirmed with mass spectrometry techniques.


Subject(s)
Electrophoresis, Gel, Two-Dimensional/methods , Immunoblotting/methods , Mass Spectrometry/methods , Mitochondria, Heart/chemistry , Mitochondrial Proteins/analysis , Peroxynitrous Acid/chemistry , Tyrosine/analogs & derivatives , Animals , Cattle , Mitochondria, Heart/metabolism , Mitochondrial Proteins/metabolism , Tyrosine/analysis , Tyrosine/metabolism
18.
ACS Appl Mater Interfaces ; 13(24): 28610-28626, 2021 Jun 23.
Article in English | MEDLINE | ID: mdl-34110776

ABSTRACT

The development of aggregation-induced emission enhancement (AIEE) active nanoprobes without any synthetic complication for solution-state and organic thin-film transistor (OTFT)-based sensory applications is still a challenging task. In this study, the novel pyrene-incorporated Schiff base (5-phenyl-4-((pyren-1-ylmethylene)amino)-4H-1,2,4-triazole-3-thiol; PT2) with an AIEE property was synthesized via a one-pot reaction and was reported for detecting Zn2+ and tyrosine in the solution state and OTFT. In the AIEE studies of PT2 (in CH3CN) at various water fractions (fw: 0-97.5%), the existence of J-aggregation, crystalline changes, and nanofibers formation was confirmed by ultraviolet absorption/photoluminescence (UV/PL) spectroscopy, powder X-ray diffraction (PXRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM), atomic force microscopy (AFM), and dynamic-light scattering (DLS) techniques. Similarly, PT2-based Zn2+ detection and sensory reversibility with tyrosine were demonstrated by UV/PL studies with evidence related to crystalline/nanolevel changes in PXRD, SEM, TEM, AFM, and DLS data. Distinct decay profiles associated with the AIEE and sensory responses of PT2 were observed in time-resolved photoluminescence spectra. From the standard deviation and linear fittings of PL titrations, detection limits (LODs) of the Zn2+ with PT2 and the tyrosine with PT2-Zn2+ were estimated as 0.79 and 45 nM, respectively. High-resolution mass and 1H NMR results confirmed 2:1 and 1:1 stoichiometry and binding sites of PT2-Zn2+-PT2* and tyrosine-Zn2+ complexes. Moreover, the values of association constants determined by linear fittings were 4.205 × 10-7 and 1.73 × 10-8 M-2, correspondingly. Optimization via the density functional theory disclosed the binding sites and suppression of twisted intramolecular charge transfer/photoinduced electron transfer (TICT/PET) as well as the involvement of restricted intramolecular rotation in the AIEE and PET "ON-OFF-ON" mechanisms in the Zn2+ and tyrosine sensors. Results from the B16-F10 cellular and zebrafish imaging of AIEE, Zn2+, and tyrosine sensors further attested the applicability of PT2 in biological samples. Finally, the PT2 and pentacene-incorporated OTFT devices were fabricated. The devices displayed more than 90% change in drain-source current when reacted with Zn2+ with an LOD of 5.46 µM but showed no response to tyrosine, thereby confirming the reversibility. Moreover, the OTFT devices also demonstrated Zn2+ ion detection in tap water and lake water samples.


Subject(s)
Fluorescent Dyes/chemistry , Pyrenes/chemistry , Tyrosine/analysis , Zinc/analysis , Animals , Density Functional Theory , Drinking Water/analysis , Electrochemical Techniques/methods , Fluorescent Dyes/chemical synthesis , Lakes/analysis , Limit of Detection , Models, Chemical , Pyrenes/chemical synthesis , Schiff Bases/chemical synthesis , Schiff Bases/chemistry , Transistors, Electronic , Zebrafish
19.
J Mass Spectrom ; 56(6): e4713, 2021 Jun.
Article in English | MEDLINE | ID: mdl-33942435

ABSTRACT

Oxidative and nitrative stress have been implicated in the molecular mechanisms underlying a variety of biological processes and disease states including cancer, aging, cardiovascular disease, neurological disorders, diabetes, and alcohol-induced liver injury. One marker of nitrative stress is the formation of 3-nitrotyrosine, or protein tyrosine nitration (PTN), which has been observed during inflammation and tissue injury; however, the role of PTN in the progression or possibly the pathogenesis of disease is still unclear. We show in a model of alcohol-induced liver injury that an increase in PTN occurs in hepatocyte nuclei within the liver of wild-type male C57BL/6J mice following chronic ethanol exposure (28 days). High-resolution mass spectrometric analysis of isolated hepatic nuclei revealed several novel sites of tyrosine nitration on histone proteins. Histone nitration sites were validated by tandem mass spectrometry (MS/MS) analysis of representative synthetic nitropeptides equivalent in sequence to the respective nitrotyrosine sites identified in vivo. We further investigated the potential structural impact of the novel histone H3 Tyr41 (H3Y41) nitration site identified using molecular dynamics (MD) simulations. MD simulations of the nitrated and non-nitrated forms of histone H3Y41 showed significant structural changes at the DNA interface upon H3Y41 nitration. The results from this study suggest that, in addition to other known post-translational modifications that occur on histone proteins (e.g., acetylation and methylation), PTN could induce chromatin structural changes, possibly affecting gene transcription processes associated with the development of alcohol-induced liver injury.


Subject(s)
Chemical and Drug Induced Liver Injury, Chronic/metabolism , Ethanol/metabolism , Histones/analysis , Nitrates/metabolism , Tyrosine/analogs & derivatives , Amino Acid Sequence , Animals , Disease Models, Animal , Histones/metabolism , Liver , Male , Mice , Mice, Inbred C57BL , Molecular Dynamics Simulation , Oxidative Stress , Tandem Mass Spectrometry , Tyrosine/analysis , Tyrosine/metabolism
20.
Virology ; 559: 111-119, 2021 07.
Article in English | MEDLINE | ID: mdl-33865074

ABSTRACT

Influenza A virus (IAV) infection alters lung epithelial cell metabolism in vitro by promoting a glycolytic shift. We hypothesized that this shift benefits the virus rather than the host and that inhibition of glycolysis would improve infection outcomes. A/WSN/33 IAV-inoculated C57BL/6 mice were treated daily from 1 day post-inoculation (d.p.i.) with 2-deoxy-d-glucose (2-DG) to inhibit glycolysis and with the pyruvate dehydrogenase kinase (PDK) inhibitor dichloroacetate (DCA) to promote flux through the TCA cycle. To block OXPHOS, mice were treated every other day from 1 d.p.i. with the Complex I inhibitor rotenone (ROT). 2-DG significantly decreased nocturnal activity, reduced respiratory exchange ratios, worsened hypoxemia, exacerbated lung dysfunction, and increased humoral inflammation at 6 d.p.i. DCA and ROT treatment normalized oxygenation and airway resistance and attenuated IAV-induced pulmonary edema, histopathology, and nitrotyrosine formation. None of the treatments altered viral replication. These data suggest that a shift to glycolysis is host-protective in influenza.


Subject(s)
Epithelial Cells/metabolism , Glycolysis , Influenza A Virus, H1N1 Subtype/pathogenicity , Lung Injury/virology , Lung/metabolism , Animals , Female , Lung/chemistry , Lung/virology , Lung Injury/prevention & control , Male , Mice , Mice, Inbred C57BL , Pyruvate Dehydrogenase Acetyl-Transferring Kinase/antagonists & inhibitors , Tyrosine/analogs & derivatives , Tyrosine/analysis , Tyrosine/metabolism , Virus Replication
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