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Surface-enhanced Raman scattering (SERS) is a powerful technique primarily due to its high sensitivity and signal-enhancing properties, which enable the identification of unique vibrational fingerprints. These fingerprints can be used for the diagnosis and monitoring of diseases such as cancer. It is crucial to selectively identify cancer biomarkers for early diagnosis. A correlation has been established between the reduction in the concentration of specific amino acids and the stage of the disease, particularly tryptophan (TPP) and tyrosine (TRS) in individuals diagnosed with prostate cancer. In this work, we present a strategy to analyze TPP and TRS amino acids using molecularly imprinted polymer nanoparticles (nanoMIPs), which selectively detect target molecules in a SERS sensor. NanoMIPs are synthesized using the solid-phase molecular imprinting method with TPP and TRS as templates. These are then immobilized on a SERS substrate with gold nanoparticles to measure samples prepared from tryptophan and tyrosine in phosphate-buffered saline. The detection and quantification limits of the designed sensor are 7.13 μM and 23.75 μM for TPP, and 22.11 μM and 73.72 μM for TRS, respectively. Our study lays the groundwork for future investigations utilizing nanoMIPs in SERS assessments of TPP and TRS as potential biomarkers for prostate cancer detection., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Pepsin as an aspartic acid protease member and one of the three foremost proteolytic enzymes in the digestive system is essential to be detected. An electrochemically polymerized tyrosine film on carbon paste electrode (pTyr/CPE) has been synthesized by electro-polymerization donating an affordable electrochemical sensor to sense salivary pepsin as a diagnostic technique for gastroesophageal reflux disease (GRD) due to saliva collection is non-invasive and relatively comfortable. The pTyr/CPE was applied for Voltammetric sensing of pepsin and its quantification in phosphate buffer solution of pH 2.0 (PBS). Scanning electron microscopy (SEM) was conducted to learn the surface morphology. Cyclic voltammetry (CV), differential pulse voltammetry (DPVs), chronoamperometry (CA), and electrochemical impedance spectroscopy (EIS) were developed to realize the electrocatalytic activity of the sensor. The pTyr/CPE proceeded as a sensitive detector to pepsin with two linear ranges from 1 to 20 & 20 to 100 ng/mL donating two limits of detection as 0.5 & 0.09 ng/mL, respectively, and high selectivity toward pepsin, as well as stability and fast response of 1.5 s. Consequently, it is guessed that the pTyr/CPE sensor could be supportive for the initial diagnosis of GRD through the detection of pepsin in saliva. Finally, we quantified the pepsin levels in saliva samples of LPR patients (n = 2), showing that the results were agreeable with those from the electrochemical sensor., Competing Interests: Declaration of competing interest no conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

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 (eMoS x ) on laser induced graphene (LIG) for multiplexed detection of tyrosine (TYR) and uric acid (UA) in sweat and saliva. Electrodeposition of MoS x shows an increased electrochemically active surface area (ECSA) and heterogeneous electron transfer rate constant, k 0 . 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 eMoS x -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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

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., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Melanin synthesis occurs within a specialized organelle called the melanosome. Traditional methods for measuring melanin levels rely on the detection of chemical degradation products of melanin by high-performance liquid chromatography. Although these methods are robust, they are unable to distinguish between melanin synthesis and degradation and are best suited to measure melanin changes over long periods of time. We developed a method that actively measures both eumelanin and pheomelanin synthesis by fate tracing [U- 13 C] L-tyrosine using liquid chromatography-mass spectrometry. Using this method, we confirmed the previous reports of the differences in melanin synthesis between melanocytes derived from individuals with different skin colors and MC1R genotype and uncovered new information regarding the differential de novo synthesis of eumelanin and pheomelanin, also called mixed melanogenesis. We also revealed that distinct mechanisms that alter melanosomal pH differentially induce new eumelanin and pheomelanin synthesis. Finally, we revealed that the synthesis of L-3,4-dihydroxyphenylalanine, an important metabolite of L-tyrosine, is differentially controlled by multiple factors. Because L-tyrosine fate tracing is compatible with untargeted liquid chromatography-mass spectrometry‒based metabolomics, this approach enables the broad measurement of cellular metabolism in combination with melanin metabolism, and we anticipate that this approach will shed new light on multiple mechanisms of melanogenesis., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

The aim of this study was to quantify the effects of different processing parameters on texture development and the incidence of white film and tyrosine crystals in dry-cured ham. Hams were dry-salted for 0.65, 0.8 or 1.0 days/kg. After drying for 45 days at 5 °C, they were dried at 10, 15 or 20 °C until reaching 33% weight loss and, thereafter, dried at 25 °C until reaching 36 or 40% weight loss. The salting time, drying temperature and target weight loss significantly affected the texture and incidence of white film and tyrosine crystals. A beneficial effect of drying at 20 °C on texture was found, which was especially important for low target weight loss (33%). Besides, hams dried at 20 °C and those with 40% weight loss showed higher incidence of tyrosine crystals. Contour plots and predictive models for texture can be used to define optimal processing parameters., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Concurrent measurement of tyrosine, tryptophan and their metabolites, and other co-factors could help to diagnose and better understand a wide range of metabolic and neurological disorders. The two metabolic pathways are closely related to each other through co-factors, regulator molecules and enzymes. By using high performance liquid chromatography coupled to electrospray ionization triple quadrupole mass spectrometry, we present a robust, selective and comprehensive method to determine 30 molecules within 20 min using a Waters Atlantis dC18. The method was validated according to the guideline of European Medicines Agency on bioanalytical method validation. Analytical performance met all the EMA requirements and the assay covered the relevant clinical concentrations. Linear correlation coefficients were all >0.998. Intra-day and inter-day accuracy were between 80-119% and 81-117%, precision 1-19% respectively. The method was applied to measure TYR, TRP and their metabolites, and other neurologically important molecules in human serum and CSF samples. The assay can facilitate the diagnosis and is suitable for determination of reference values in clinical laboratories., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:, (Copyright © 2020. Published by Elsevier B.V.)

Background: Chemokine receptor tyrosine sulfation plays a key role in the binding of chemokines. It has been suggested that receptor sulfation is heterogeneous, but no experimental evidence has been provided so far. The potent anti-HIV chemokine analog 5P12-RANTES has been proposed to owe its inhibitory activity to a capacity to bind a larger pool of cell surface CCR5 receptors than native chemokines such as CCL5, but the molecular details underlying this phenomenon have not been elucidated., Methods: We investigated the CCR5 sulfation heterogeneity and the sensitivity of CCR5 ligands to receptor sulfation by performing ELISA assays on synthetic N-terminal sulfopeptides and by performing binding assays on CCR5-expressing cells under conditions that modulate CCR5 sulfation levels., Results: Two commonly used anti-CCR5 monoclonal antibodies with epitopes in the sulfated N-terminal domain of CCR5 show contrasting binding profiles on CCR5 sulfopeptides, incomplete competition with each other for cell surface CCR5, and opposing sensitivities to cellular treatments that affect CCR5 sulfation levels. 5P12-RANTES is less sensitive than native CCL5 to conditions that affect cellular CCR5 sulfation., Conclusions: CCR5 sulfation is heterogeneous and this affects the binding properties of both native chemokines and antibodies. Enhanced capacity to bind to CCR5 is a component of the inhibitory mechanism of 5P12-RANTES., General Significance: We provide the first experimental evidence for sulfation heterogeneity of chemokine receptors and its impact on ligand binding, a phenomenon that is important both for the understanding of chemokine cell biology and for the development of drugs that target chemokine receptors., (Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.)

The prominent biological effects of adrenaline (A), noradrenaline (NA) and dopamine (DA) as well as the clinical importance of their metabolites (such as dihydroxyphenylacetic acid (DOPAC), methoxy‑4-hydroxyphenyl glycol (MHPG), dihydroxyphenylglycol (DHPG), metanephrine (M), normetanephrine (NM), vanillylmandelic acid (VMA), homovanillic acid (HVA)) have forced researchers to evaluate new analytical methodologies for their isolation and preconcentration from biological samples. For this reason, the three most popular extraction techniques (dispersive liquid-liquid microextraction (DLLME), solid-phase extraction (SPE), solid-phase microextraction (SPME)) were tested. Micellar electrokinetic chromatography (MEKC) - a mode of capillary electrophoresis - with a diode array detector (DAD) was applied to assess the extraction efficiency. Next, the enrichment factor (EF) of each applied method was calculated in respect to standard mixtures of the analytes at the same concentration levels. The EF results of seven selected metabolites of biogenic amines (BAs) from urine after sample preparation procedures based on twenty-five different protocols (one DLLME, thirteen SPE and eleven SPME) were calculated and compared using hierarchical cluster analysis (HCA). The SPE as well as SPME procedures were proved to be the most effective approaches for the simultaneous extraction of the chosen compounds. Moreover, an ionic liquid (IL) - 1-ethyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide - added to methanol in SPME additionally could successfully improve the extraction efficiency. It was also confirmed that the HCA approach could be considered a supportive tool in the selection of a suitable sample preparation procedure for that group of endogenous substances., (Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.)

Polymers obtained from tetraethyl orthosilicate (TEOS) and triethoxymethylsilane (MTEOS) have been functionalized with different metal and metal oxide nanoparticles (NPs), and used as coatings of extractive capillaries for the extraction of polar compounds by in-tube solid-phase microextraction (IT-SPME) coupled on-line to nano-liquid chromatography (nano-LC). The extraction capabilities of the new phases have been studied using several triazinic herbicides with log of octanol/water partition coefficients (K ow ) ranging from -0.7 to 3.21 under reversed phase chromatographic conditions. Best extraction efficiencies for the most polar compounds (log K ow ≤ 2.3) were typically obtained with the CuO NPs doped phase. The TEOS-MTEOS polymer can be modified with two types of NPs in order to obtain extractive phases capable of interacting with compounds of a wide range of polarities; alternatively, two capillaries each with a different type of NPs can be combined in series with the same goal. Under the later approach the limits of detection (LODs) found for the tested herbicides were 0.02-1.5 µg/L, and the precision expressed as relative standard deviation (RSD) varied from 2 to 10% (n = 3). The recoveries found in sea water samples ranged from 80 to 107%. In addition, the developed CuO NPs doped phase can be used in hydrophilic interaction chromatography (HILIC), which is the separation mode recommended for highly polar compounds. This has been illustrated using the amino acids tyrosine (log K ow  = -2.26) and tryptophan (log K ow  = -1.06) as model compounds, being their respective LOD 0.1 and 0.3 µg/mL. Examples of application of the developed bimodal extractive phase to different environmental and waste waters are given in order to show its utility and versatility., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2019. Published by Elsevier B.V.)

A calix[4]arene based compound incorporating amide and morpholine moieties has been synthesized and its ion recognition property towards metal ions and application of its metal complex towards sensing of amino acids has been investigated. The synthesized compound interacts with Cu 2+ with high selectivity and sensitivity (LOD, 0.1 ppb) in aqueous media with instant color change from colorless to yellow without interference from any other metal ions used in this study. The molecular structure of the calix compound (1) has been determined by single crystal X-ray study and the structure of its Cu 2+ complex has been established by DFT calculation. The Cu 2+ complex of 1 selectively detects tyrosine (LOD, 1.2 ppm) in water with distinct color change and without any interference from other 22 amino acids used in this study. The mechanism for detection of tyrosine with color change is also presented. For easy field application, paper based sensor strips have been prepared by coating compound 1 and its Cu 2+ complex on filter paper, which have been used for semi-quantitative measurement of Cu 2+ and tyrosine. Compound 1 and its Cu 2+ complex have also been used for detection of Cu 2+ and tyrosine, respectively in water and human saliva as real samples and satisfactory results are obtained., Competing Interests: Declaration of competing interest The authors declare no competing financial interest. The authors also declare no conflict of interest in this work., (Copyright © 2020 Elsevier B.V. All rights reserved.)

A polyaminosaccharide (chitosan, CS) and an aminosaccharide (d-galactosamine, GalN) were integrated together via hydrothermal assembly to obtain a bis-aminosaccharides composite (CS-GalN), and a novel and facile chiral sensing platform based on CS-GalN modified glassy carbon electrode (CS-GalN/GCE) was fabricated and used for electrochemical recognition of tyrosine (Tyr) enantiomers. CS-GalN composite was characterized by Fourier transform infrared (FT-IR) spectroscopy, scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDS) and contact angle goniometry. It was observed that CS-GalN composite exhibited different binding ability for Tyr enantiomers. Under optimized experimental conditions, the oxidation peak current ratio of L-Tyr to D-Tyr (I L /I D ) and the difference between their peak potentials (ΔEp = E D -E L ) were 1.70 and 28 mV at CS-GalN/GCE by square wave voltammetry (SWV). In addition, the peak currents increase linearly with the concentration of Tyr enantiomers in the concentration range 0.01-1.00 mM with detection limits of 0.65 μM and 0.86 μM for L-Tyr and D-Tyr (S/N = 3), respectively. CS-GalN/GCE also exhibited the ability to determine the percentage of D-Tyr in the racemic mixture. In addition, CS-GalN/GCE possessed remarkable sensitivity, great stability as well as fine reproducibility. It could be concluded that the chiral interface of CS-GalN/GCE can provide an ideal platform for electrochemical recognition and determination of Tyr enantiomers., (Copyright © 2019 Elsevier B.V. All rights reserved.)

In this work, molecularly imprinted electrospun chromogenic membrane(MIP-ECM) was synthesized using P-Hydroxybenzene propanoic acid as imitate template molecule and Poly (vinyl alcohol)(PVA) as substrate. Ninhydrin is used as chromogenic agent for visualizing detection of target molecule l-tyrosine(Tyr). The results showed that molecularly imprinted membrane exhibited selectivity and specific adsorption capacity for l-tyrosine. The influences of environmental factors, such as reaction temperature, the amount of ninhydrin and elution time, on chromogenic properties were examined by SEM and optical microscope. Gray value analysis is proceeded by software Image J. Due to the facilitate preparation and intuitive visualization, the proposed method was a promising candidate for selective recognition of bio-generated acid and amino molecules., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: In abdominal aortic aneurysm (AAA), pathophysiology deterioration of the medial aortic layer plays a critical role. Key players in vessel wall degeneration are reactive oxygen species (ROS), smooth muscle cell apoptosis, and extracellular matrix degeneration by matrix metalloproteinase-9 (MMP-9). Lipocalin-2, also neutrophil gelatinase-associated lipocalin (NGAL), is suggested to be involved in these degenerative processes in other cardiovascular diseases. We aimed to further investigate the role of NGAL in AAA development and rupture., Methods: In this observational study, aneurysm tissue and blood of ruptured (n = 13) AAA patients were investigated versus nonruptured (n = 26) patients. Nondilated aortas (n = 5) from deceased patients and venous blood from healthy volunteers (n = 10) served as controls. NGAL concentrations in tissue and blood were measured by enzyme-linked immunosorbent assay and immunofluorescence microscopy. Nitrotyrosine (marker of ROS), MMP-9, and caspase-3 (marker of apoptosis) in aneurysm tissue were measured by immunofluorescence microscopy. AAA expansion rates were calculated retrospectively., Results: NGAL (in μg/mL) blood concentration in ruptured AAA was 46 (range 22-122) vs. 26 (range 6-55) in nonruptured AAA (P < 0.01) and 14 (range 12-22) in controls (P < 0.01). In the aneurysm wall of ruptured AAA, NGAL concentration was 4.7 (range 1.4-25) vs. 4.4 (range 0.2-14) in nonruptured AAA (not significant) and 1.8 (range 1.2-2.7) in nondilated aortas (P = 0.04). In the medial layer, NGAL correlated positively with nitrotyrosine (Rs = 0.80, P < 0.01), MMP-9 (Rs = 0.56, P = 0.02), and caspase-3 (Rs = 0.75, P = 0.01). NGAL did not correlate to AAA expansion rate in blood or tissue (P = 0.34 and P = 0.95, respectively)., Conclusions: This study demonstrates that NGAL blood concentration is higher in ruptured AAA patients than in nonruptured AAA. NGAL expression in the AAA wall is also higher than in nondilated aorta. Furthermore, its expression is associated with factors of vessel wall deterioration. Based on our study results, we could not determine NGAL as a biomarker for AAA growth or rupture. However, our findings do support a potential role of NGAL in the development of AAA., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Dysregulation and aggregation of the peptide hormone IAPP (islet amyloid polypeptide, a.k.a. amylin) into soluble oligomers that appear to be cell-toxic is a known aspect of diabetes mellitus (DM) Type 2 pathology. IAPP aggregation is influenced by several factors including interactions with metal ions such as Cu(II). Because Cu(II) ions are redox-active they may contribute to metal-catalyzed formation of oxidative tyrosyl radicals, which can generate dityrosine cross-links. Here, we show that such a process, which involves Cu(II) ions bound to the IAPP peptide together with H 2 O 2 , can induce formation of large amounts of IAPP dimers connected by covalent dityrosine cross-links. This cross-linking is less pronounced at low pH and for murine IAPP, likely due to less efficient Cu(II) binding. Whether IAPP can carry out its hormonal function as a cross-linked dimer is unknown. As dityrosine concentrations are higher in blood plasma of DM Type 2 patients - arguably due to disease-related oxidative stress - and as dimer formation is the first step in protein aggregation, generation of dityrosine-linked dimers may be an important factor in IAPP aggregation and thus relevant for DM Type 2 progression., (Copyright © 2019 Elsevier Inc. All rights reserved.)

A new high sensitivity deep-UV LED photometric detector with a z-type flow cell (45 nL or 180 nL) for miniaturised and portable capillary liquid chromatography (LC) was designed and fabricated to overcome sensitivity limitations due to short pathlength in on-capillary detectors. The new detector has a 10 mm geometric pathlength and uses high intensity light-emitting diodes (LED) as light sources in the deep-UV range (254 nm and 280 nm). No optical reference was necessary due to the low drift in the signal. Stray light was minimized by the use of an adjustable slit with a 0.5 mm pinhole. The direct relationship between absorbance and concentration was obtained using dichromate to evaluate the sensitivity and the linearity range of the detector. Performance of the miniaturised version was compared with that obtained from a commercial benchtop detector for capillary LC under the same conditions using the same optical z-cell. The miniaturised version exhibited a superior performance across all parameters, including 3 times higher effective pathlength, 4 times higher upper limit of detector linearity, and 2-5 times lower stray light levels. An application of the new detector was shown with the detection of l-dopa, l-tyrosine, norfenefrine, phenylephrine and tyramine, separated using capillary LC. The baseline noise level recorded was as low as 3.9 μAU. Further, the detector was applied in a miniaturised capillary LC for the indirect detection of common inorganic anions. In comparison to an on-capillary LED detector applied under similar chromatographic conditions, there was a 50 times higher signal to noise (S/N) ratio., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Here, we aimed to use graphene oxide to improve the selectivity and sensitivity of Tyr determination via the reaction with 1-nitroso-2-naphthol as a selective reagent of Tyr. The reaction between Tyr and 1-nitroso-2-naphthol in absence and presence of GO was studied spectrophotometrically. Different parameters such as concentrations, temperature, incubation time were optimized. The obtained data showed that the maximum absorbance was achieved by using 2 mL of 0.03% 1-nitroso-2-naphthol at temperature 60 °C for 10 min. On the basis of calibration curve of various concentrations of Tyr in the presence of 20 μg mL -1 GO, the limit of detection was 6.4 × 10 -6  M (1.15 μg mL -1 ), where in absence of GO was 1.1 × 10 -5  M (19.9 μg mL -1 ). The selectivity of Tyr in presence of other amino acids and phenols was studied with and without GO. The data obtained revealed that the selectivity of Tyr in presence of GO with respect to some amino acids and phenols was improved. The proposed method has been applied for the determination of Tyr in urine and serum samples. Therefore, GO is a powerful catalytic surface for the sensitive and selective determination of Try in biological fluids., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The tyrosine phosphorylation of proteins and peptides plays a vital role in cell signal transduction pathways, and it is very important to assay them for understanding their action mechanism. Due to the low levels of the tyrosine phosphopeptides (pTyr) in cells, it is a challenge to enrich them with traditional sorbents, therefore, development of specific and selective sorbents is urgent and necessary. In this work, the phosphate-imprinted magnetic nanoparticles (PMNPs) to enrich the pTyr with high efficiency and selectivity have been fabricated using the phenylphosphonic acid as a template for the "epitope" of pTyr. The magnetic nanoparticles have been functionalized with TiO 2 and then the imprinting silica shells have been coated on the surface of the functional core to obtain the PMNPs sorbents. The PMNPs can obviously shorten the enrichment time and improve the adsorption efficiency for pTyr, and the epitope imprinting films provide an excellent selective recognition ability to target. The recognition capability of PMNPs for pTyr is 90.3 μg/mg and the imprinting factor of the sorbents can reach 24.4. The results indicate that the PMNPs can enrich the pTyr from the tryptic digest of β-casein samples with high specificity, and the spiking recoveries of the pTyr range from 85.1% to 93.8% with the RSD from 0.04 to 3.73. With the high adsorption capacity, rapid separation, excellent specificity and recyclability, the PMNPs sorbents show great potential for analysis of the phosphorylation of peptides in biological and medical fields., (Copyright © 2018 Elsevier B.V. All rights reserved.)

In this paper, a new and facile method for the electrochemical determination of l-tyrosine was designed. First, 3-mercaptopropyl trimethoxysilane-functionalized silica nanoparticles were added to a paper disc. Then, the banana peel tissue and the mediator potassium hexacyanoferrate were dropped onto the paper, respectively. The modified paper disc was placed on the top of the graphite screen printed electrode and electrochemical characterization of this biosensor was studied by cyclic voltammetry and electrochemical impedance spectroscopy methods. The effective parameters like pH, banana peel tissue percentage, and the amount of mediator loading were optimized. l-tyrosine measurements were done by differential pulse voltammetry with a little sample (3 μL) for analysis. The biosensor showed a linear response for l-tyrosine in the wide concentration range of 0.05-600 μM and a low detection limit about 0.02 μM because of the co-catalytic effect of enzyme and nanoparticles. The stability of the biosensor and its selectivity were evaluated. This biosensor was applied for the voltammetric determination of l-tyrosine in the blood plasma sample. The results of the practical application study were comparable with the standard method (HPLC). In conclusion, a simple, inexpensive, rapid, sensitive and selective technique was successfully applied to the l-tyrosine analysis of the little samples., (Copyright © 2018 Elsevier B.V. All rights reserved.)

In this work, fabrication of a novel and ultrasensitive electrochemical biosensor based on immobilization of tyrosine hydroxylase onto palladium-platinum bimetallic alloy nanoparticles/chitosan-1-ethyl-3-methylimidazolium bis(trifluoromethylsulfonyl) imide/graphene-multiwalled carbon nanotubes-IL/glassy carbon electrode for determination of L-tyrosine in some high tyrosine foods including cheese, egg and yogurt was reported. Immobilization of tyrosine hydroxylase onto the surface of the biosensor was performed by cross-linking tyrosine hydroxylase and chitosan through the addition of glutaraldehyde. Enzymatic biosensors employ the affinity and selectivity of catalytically active proteins towards their target molecules and here, the tyrosine hydroxylase selectively catalyzes the conversion of tyrosine to levodopa which can be oxidized at lower potentials than tyrosine. The modifications were characterized by electrochemical impedance spectroscopy, cyclic voltammetry, energy dispersive X-ray spectroscopic and scanning electron microscopy. Under optimal conditions, the biosensor detected tyrosine in concentration ranges of 0.01 × 10 -9 to 8.0 × 10 -9 mol L -1 and 8.0 × 10 -9 to 160.0 × 10 -9 mol L -1 with a limit of detection of 0.009 × 10 -9 mol L -1 . The biosensor was able to selective determination of tyrosine even in the presence of common interferents therefore, the biosensor was highly selective. The biosensor also showed good operational stability, antifouling properties, sensitivity, repeatability and reproducibility., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Protein tyrosine sulfation (Tyr-O-SO3) is a common post-translational modification (PTM), which is important for protein function. Absolute quantitation of Tyr-O-SO3 in recombinant therapeutic proteins has been challenging. We report here an MRM method used for absolute quantitation of Tyr-O-SO3 in the hydrolysate of a recombinant Fc-fusion protein. Quantitation is achieved by monitoring the sum of two transitions: the loss of carboxylic acid from tyrosine sulfate (major transition) and sulfate group from tyrosine sulfate sodium salt. The method exhibits a good sensitivity with a limit of quantitation of 1.4 ng/mL, linearity over three orders of magnitude, good repeatability, precision and accuracy., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Tyro3, a member of TAM receptor tyrosine kinase family has been suggested to be autophosphorylated upon activation. In the current study we mapped the autophosphorylation sites of murine Tyro3 to tyrosine 723 and 756, with K540 being required for its kinase activity. Knockdown of Axl significantly decreases the tyrosyl-phosphorylation of Tyro3 in fibroblasts NR6WT, suggesting an interaction among the TAM family members. Interestingly, the carboxyl terminal region of Tyro3 is required for its stability in cells with a minimal length of 1-778 amino acids which is not conserved in murine Axl, a member of TAM. These data suggest that the autophosphorylation sites of TAM RTK members are unique although they share high similarity in amino acids within their carboxyl kinase domain., (Copyright © 2017 Elsevier Inc. All rights reserved.)

To explain the formation of protein nitration marker, 3 nitrotyrosine (3NT), in food products was the basis for the present study. For the accurate detection as protein nitration marker of 3NT in food products, i) a new method was improved, which is optimized parameters affected the formation of 3NT, ii) HPLC-DAD was applied as a straightforward and rapid methodology and iii) finally the suspensions representing meat and dairy products were prepared and this method was used to these suspensions to evaluate the convenience of it. For comparative purposes, the spectrophotometric method of the determination of 3NT level was also employed to assess the 3NT level. The results showed that optimized method enabled detection of 3NT level in food systems and 3NT could act as a protein nitration biomarker in food. This study will shed light on the understanding of precise mechanisms about protein nitration and contribute to the change of amino acids during processing and storage., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Tyrosine and tyrosine residues in proteins are attacked by the reactive oxygen and nitrogen species peroxynitrite (O=N-OO - ) to generate 3-nitrotyrosine (3-NT) and 3-nitrotyrosine-proteins (3-NTProt), respectively. 3-NT and 3-NTProt are widely accepted as biomarkers of nitr(os)ative stress. Over the years many different analytical methods have been reported for 3-NT and 3-NTProt. Reported concentrations often differ by more than three orders of magnitude, indicative of serious analytical problems. Strategies to overcome pre-analytical and analytical shortcomings and pitfalls have been proposed. The present review investigated whether recently published work on the quantitative measurement of biological 3-nitrotyrosine did adequately consider the analytical past of this biomolecule. 3-Nitrotyrosine was taken as a representative of biomolecules that occur in biological samples in the pM-to-nM concentration range. This examination revealed that in many cases the main protagonists involved in the publication of scientific work, i.e., authors, reviewers and editors, failed to do so. Learning from the analytical history of 3-nitrotyrosine means advancing analytical and biological science and implies the following key issues. (1) Choosing the most reliable analytical approach in terms of sensitivity and accuracy; presently this is best feasible by stable-isotope dilution tandem mass spectrometry coupled with gas chromatography (GC-MS/MS) or liquid chromatography (LC-MS/MS). (2) Minimizing artificial formation of 3-nitrotyrosine during sample work up, a major pitfall in 3-nitrotyrosine analysis. (3) Validating adequately the final method in the intendent biological matrix and the established concentration range. (4) Inviting experts in the field for critical evaluation of the novelty and reliability of the proposed analytical method, placing special emphasis on the compliance of the analytical outcome with 3-nitrotyrosine concentrations obtained by validated GC-MS/MS and LC-MS/MS methods., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Tabun has been shown to form phosphylated adducts on tyrosine residues in albumin in vivo and in vitro. However, in this work, tabun-labeled lysine adducts were found in albumin. Three types of albumin were treated with overdose of tabun in vitro and 17 tabun-labeled lysine residues were found: K4, K12, K224, K377, and K524 in bovine albumin, K186, K188, K212, K329, K414, and K525 in leporine albumin, and K79, K186, K188, K212, K376, and K525 in rat albumin. To investigate the modification of tabun in vivo, three leporines were injected with 0.8×LD 50 dose of tabun. The results showed that the labeled lysine residues in vivo, were consistent with modified lysines in vitro. Structure characteristics and the binding mode of 6 tabun-labeled lysines of leporine albumin were further analyzed using theory simulation and molecular docking in Discovery Studio. For the first time, we show that tabun-labeled lysine peptides are found in vivo and in vitro. These modified lysine peptides are good biomarkers for exposure to tabun in albumin of leporine and rat., (Copyright © 2017 Elsevier B.V. All rights reserved.)

A novel nanomaterial-based voltammetric sensor has been developed for use a highly sensitive tool for the simultaneous determination of captopril (CA), acetaminophen (AC), tyrosine (TY) and hydrochlorothiazide (HCTZ). The device is based on the application of NiO/CNTs and (2-(3,4-dihydroxyphenethyl)isoindoline-1,3-dione) (DPID) to modify carbon paste electrodes. The NiO/CNTs nanocomposite was synthesized through a direct chemical precipitation approach and was characterized with X-ray powder diffraction (XRD), and scanning electron microscopy (SEM). The NiO/CNTs/DPID/CPEs were found to facilitate the analysis of CA, AC, TY and HCTZ in the concentration ranges of 0.07-200.0, 0.8-550.0, 5.0-750.0 and 10.0-600.0μM with the respective detection limits of 9.0nM, 0.3μM, 1.0μM and 5.0μM. The developed NiO/CNTs/DPID/CPEs were used for the determination of the mentioned analytes in pharmaceutical and biological real samples., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The chemical degradation of farglitazar (1) was investigated using a series of controlled stress testing experiments. Farglitazar drug substance was stressed under acidic, natural pH, basic, and oxidative conditions in solution. In the solid state, the drug substance was stressed with heat, high humidity, and light. Farglitazar was found to be most labile toward oxidative stress. A series of mechanistic experiments are described in which the use of 18 O-labelled oxygen demonstrated that oxidative degradation of farglitazar is caused primarily by singlet oxygen formed under thermal conditions. Major degradation products were isolated and fully characterized. Mechanisms for the formation of degradation products are proposed. Drug product tablets were also stressed in the solid state with heat, high humidity, and light. Stressed tablets afforded many of the same degradation products observed during drug substance stress testing, with oxidation again being the predominant degradation pathway. Evidence for the activity of singlet oxygen, formed during thermal stress testing of the solid oral dosage form, is presented. The degradation pathways observed during stress testing matched those observed during long-term stability trials of the drug product., (Copyright © 2017 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.)

Apixaban and Tirofiban Hydrochloride are low molecular weight anticoagulants. The two drugs exhibit native fluorescence that allow the development of simple and valid spectrofluorimetric methods for the determination of Apixaban at λ ex/λ em=284/450nm and tirofiban HCl at λ ex/λ em=227/300nm in aqueous media. Different experimental parameters affecting fluorescence intensities were carefully studied and optimized. The fluorescence intensity-concentration plots were linear over the ranges of 0.2-6μgml -1 for apixaban and 0.2-5μgml -1 for tirofiban HCl. The limits of detection were 0.017 and 0.019μgml -1 and quantification limits were 0.057 and 0.066μgml -1 for apixaban and tirofiban HCl, respectively. The fluorescence quantum yield of apixaban and tirofiban were calculated with values of 0.43 and 0.49. Method validation was evaluated for linearity, specificity, accuracy, precision and robustness as per ICH guidelines. The proposed spectrofluorimetric methods were successfully applied for the determination of apixaban in Eliquis tablets and tirofiban HCl in Aggrastat intravenous infusion. Tolerance ratio was tested to study the effect of foreign interferences from dosage forms excipients. Using Student's t and F tests, revealed no statistically difference between the developed spectrofluorimetric methods and the comparison methods regarding the accuracy and precision, so can be contributed to the analysis of apixaban and tirofiban HCl in QC laboratories as an alternative method., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Background: The nitration of tyrosine residues in proteins is associated with nitrosative stress, resulting in the formation of 3-nitrotyrosine (3-NT). 1 3-NT levels in biological samples have been associated with numerous physiological and pathological conditions. Hence several attempts have been made in order to develop methods that accurately quantify 3-NT in these matrices. The aim of this study was to develop a simple, rapid, low-cost and sensitive high-performance liquid chromatography (HPLC)-based 3-NT quantification method., Methods: All experiments were performed on an Hitachi LaChrom Elite ® HPLC system. The method was validated according to International Conference on Harmonisation (ICH) guidelines for serum samples. Additionally, other biological matrices were tested, namely whole blood, urine, B16 F-10 melanoma cell line, growth medium conditioned with the same cell line, bacterial and yeast suspensions., Results: From all the protocols tested, the best results were obtained using 0.5% CH 3 COOH:MeOH:H 2 O (15:15:70) as mobile phase, with detection at wavelengths 215, 276 and 356nm, at 25°C, and using a flow rate of 1mLmin -1 . By using this protocol, it was possible to obtain a linear calibration curve, limits of detection and quantification in the order of μgL -1 , and a short analysis time (<15min per sample). The developed protocol allowed the successful detection and quantification of 3-NT in all biological matrices tested, with detection at 356nm., Conclusion: This method, successfully developed and validated for 3-NT quantification, is simple, cheap and fast. These features render this method a suitable option for analysis of a wide range of biological matrices, being a promising useful tool for both research and diagnosis activities., (Copyright © 2017 Elsevier B.V. All rights reserved.)

An Au-Pd/reduced graphene oxide composite was employed as a novel electrode material for the sensitive and simultaneous determination of ascorbic acid, acetaminophen and tyrosine. The electrochemical response characteristics of the modified electrode toward the analytes were investigated by differential pulse voltammetry and cyclic voltammetry. The responses of the electrochemical sensor for the target analytes were found to be improved significantly in comparison with those obtained using a conventional carbon paste electrode (CPE) and reduced graphene oxide/CPE. The experimental conditions for simultaneous determination of these species have been established. Ternary mixtures of analytes can be determined in the ranges of 0.03-9.50μM. Under optimal conditions, the limits of detection were 15.7, 7.6 and 11.1nM for ascorbic acid, acetaminophen, and tyrosine, respectively. The method was applied successfully to determine the analytes in urine, serum and pharmaceutical samples simultaneously., (Copyright © 2016 Elsevier B.V. All rights reserved.)

A simple and highly sensitive surface-enhanced resonance Raman scattering (SERRS)-based approach coupled with azo coupling reaction has been put forward for quantitative analysis of histidine and tyrosine. The SERRS-based assay is simple and rapid by mixing the azo reaction products with silver nanoparticles (AgNPs) for measurements within 2min. The limits of detection (LODs) of the method are as low as 4.33×10(-11) and 8.80×10(-11)M for histidine and tyrosine, respectively. Moreover, the SERRS fingerprint information specific to corresponding amino acids guarantees the selective detection for the target histidine and tyrosine. The results from serum indicated the potential application of the proposed approach into biological samples. Compared with the methods ever reported, the main advantages of this methodology are simpleness, rapidity without time-consuming separation or pretreatment steps, high sensitivity, selectivity and the potential for determination of other molecules containing imidazole or phenol groups., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Analysis of L-tryptophan (Trp) in biological samples has great importance for biomedical studies. Amino acid tyrosine (Tyr) that usually coexist with Trp in biological fluids can significantly interfere with reliable determination of Trp. In the current study, we demonstrate the development of two ultra-sensitive electrochemical sensor and label-free aptasensor for selective analysis of Trp in biological samples (i.e., cow's milk and human plasma, saliva and urine samples). In addition, without using AgCl/KCl, an Ag pseudo-reference screen printed electrode (Ag-PR-SPE) was exploited as a reference electrode. To prepare the engineered Trp sensor/aptasensor, a gold SPE was first modified with multiwall carbon nanotube (MWCNT-AuSPE) and then armed with Trp aptamer molecules (Apt-MWCNT-AuSPE). The prepared sensors were characterized using constant current-potentiometric stripping analysis (CC-PSA) and electrochemical impedance spectroscopy (EIS). The MWCNT-AuSPE and Apt-MWCNT-AuSPE were compared with respect to the linear detection range, limit of detection (LOD), accuracy, precision, repeatability. MWCNT-AuSPE and Apt-MWCNT-AuSPE demonstrate fast near-Nernstian response for PSA of Trp over the concentration ranging from 1.0 × 10(-9) to 2.0 × 10(-4) mol L(-1) and 1.0 × 10(-11) to 1.0 × 10(-4) mol L(-1) with detection limits of 3.6 × 10(-10) mol L(-1) and 4.9 × 10(-12) mol L(-1), respectively. Common interfering species present in the biological fluids (i.e., tyrosine, uric acid, ascorbic acid) showed no effects on the determination of Trp using CC-PSA. MWCNT-AuSPE and Apt-MWCNT-AuSPE represented well reproducibility and great precision with relative standard deviation (RSD) of 2.9% and 5.3% respectively. In comparison with the MWCNT-AuSPE, Apt-MWCNT-AuSPE provided higher sensitivity, selectivity and accuracy of Trp detection in real samples. Based on these findings, we propose the developed Apt-MWCNT-AuSPE as a simple detection method for analysis of Trp in biological samples., (Copyright © 2015 Elsevier B.V. All rights reserved.)

A novel Au nanoparticles/poly(E)-4-(p-tolyldiazenyl)benzene-1,2,3-triol (AuNPs/PTAT) film modified glassy carbon electrode (AuNPs/PTAT/GCE) was fabricated for the simultaneous determination of three antioxidants named, cysteine (Cys), uric acid (UA) and tyrosine (Tyr). The bare glassy carbon electrode (GCE) fails to separate the oxidation peak potentials of these molecules, while PTAT film modified electrode can resolve them. Electrochemical impedance spectroscopy (EIS) study indicates that the charge transfer resistance of bare electrode increased as (E)-4-(p-tolyldiazenyl)benzene-1,2,3-triol was electropolymerized at the bare electrode. Furthermore, EIS exhibits enhancement of electron transfer kinetics between analytes and electrode after electrodeposition of Au nanoparticles. Differential pulse voltammetry results show that the electrocatalytic current increases linearly in the ranges of 2-540μmolL(-1) for Cys, 5-820μmolL(-1) for UA and 10-560μmolL(-1) for Tyr with detection limits (S/N=3) of 0.04μmolL(-1), 0.1μmolL(-1) and 2μmolL(-1) for Cys, UA and Tyr, respectively. The proposed method was successfully applied for simultaneous determination of Cys, UA and Tyr in human urine samples., (Copyright © 2015 Elsevier B.V. All rights reserved.)

A simple, selective and sensitive sensor based on mesoporous silica nanoparticles modified carbon paste electrode (MSNs/CPE) is introduced for electrochemical determination of tyrosine (Tyr) and tryptophan (Trp). Compared with the unmodified electrode and commercial SiO2 modified electrode (SiO2/CPE), the oxidation peak current significantly improved for both amino acids. Under optimized experimental conditions, the oxidation peak current of Trp was linear over a concentration range of 5.0 × 10(-8) to 4.0 × 10(-4) M with a detection limit of 3.4 × 10(-8) M. The oxidation peak current of Tyr was linear over a concentration range from 5.0 × 10(-7) to 6.0 × 10(-4) M with a detection limit of 1.5 × 10(-7) M. For simultaneous determination Trp and Tyr, H-point standard addition method was applied to resolve the overlapping of differential pulse voltammetric peaks of Trp and Tyr. The results showed that the method was successfully applied to the simultaneous determination of Trp and Tyr in some synthetic samples. Moreover, the applicability of the method was demonstrated by the recovery tests of Trp and Tyr in artificial urine., (Copyright © 2015. Published by Elsevier B.V.)

The developments in the use of plasmid DNA (pDNA) in gene therapy and vaccines have motivated the search and improvement of optimized purification processes. In this context, dipeptides l-tyrosine-l-tyrosine and l-tyrosine-l-arginine are synthetized to explore their application as affinity ligands for supercoiled (sc) plasmid DNA (pDNA) purification. The synthesis is based on the protection of N-Boc-l-tyrosine, followed by condensation with l-tyrosine or l-arginine methyl esters in the presence of dicyclohexylcarbodiimide (DCC), which after hydrolysis and acidification give the afforded dipeptides. The supports are then obtained by coupling l-tyrosine, l-tyrosine-l-tyrosine and l-tyrosine-l-arginine to epoxy-activated Sepharose and are characterized by high resolution magic angle spinning (HR-MAS) NMR and Fourier transform infrared spectroscopy (FTIR). Surface plasmon resonance (SPR) biosensor is used to establish the promising ligand to be used in the chromatographic experiments and ascertain experimental conditions. Sc isoform showed the highest affinity to the dipeptides, followed by linear (ln) pDNA, being the open circular (oc) the one that promoted the lowest affinity to l-tyrosine-l-arginine. Saturation transfer difference (STD)-NMR experiments show that the interaction is mainly hydrophobic with the majority of the 5'-mononucleotides, except for 5'-GMP with l-tyrosine-l-arginine Sepharose that is mainly electrostatic. The support l-tyrosine Sepharose used in chromatographic experiments promotes the separation of native pVAX1-LacZ and pcDNA3-FLAG-p53 samples (oc+sc) by decreasing the salt concentration. The results suggest that it is possible to purify different plasmids with the l-tyrosine Sepharose, with slight adjustments in the gradient conditions., (Copyright © 2015 Elsevier B.V. All rights reserved.)

A new absorbance detector for capillary electrophoresis featuring relatively high intensity light-emitting diodes as radiation sources and photodiodes for the deep-UV range was developed. The direct relationship of absorbance values and concentrations was obtained by emulating Lambert-Beer's law with the application of a beam splitter to obtain a reference signal and a log-ratio amplifier circuitry. The performance of the cell was investigated at 255 nm with the detection of sulfanilic, 4-nitrobenzoic, 4-hydroxybenzoic and 4-aminobenzoic acid and the indirect detection of acetate, propionate, butyrate and caproate using benzoate as the displacement dye molecule. Vanillic acid, L-tyrosine and DL-tryptophan as well as the sulfonamides sulfamerazine, sulfathiazole and sulfamethazine were determined at 280 nm. Good linearities over 3 orders of magnitude were obtained. The noise level recorded was as low as 50 μAU and the drift typically <200 μAU/5 min., (Copyright © 2015 Elsevier B.V. All rights reserved.)

A new method based on photochemical formation of silver nanoparticles (AgNPs) was developed for detection of tyrosine (Tyr). To selectively detect Tyr and to simplify the detection procedure, the photoactivity of Tyr was utilized to trigger the photochemical reduction in production of AgNPs. The drastic change of solution color caused by the surface plasmon resonance (SPR) absorption band of the formed AgNPs was used to extract the quantitative information of Tyr. This developed method is simple in detection, while both the sensitivity and selectivity are significant improved. Meanwhile, the solution color was changed from colorless to dark yellow after the formation of AgNPs, which allows a much higher sensitivity in visual identification when compared with the SPR band shifting technique commonly, used in conventional colorimetric methods. To optimize the detection system and to understand the mechanism in this proposed method, parameters such as irradiation time, intensity of light source, and the concentration of Tyr were systematically examined. Results indicated that these factors mainly affected the reaction rate of photoreduction. The morphologies of the formed AgNPs were similar, but with small differences in particle sizes. In the examination of selectivity, sixteen other amino acids were examined. Results indicated that only amino acids of tryptophan, cysteine and histidine are photoactive and possess potential interferences in analysis of Tyr. Quantitative studies indicated that a linear response up to 10 μM with a detection limit of 100 nM could be obtained. For visually detection, color change could be observed with a concentration as low as 500 nM of Tyr., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The mammalian protein Girdin interacts with several key molecules such as actin, and it functions as a regulator of the cytoskeleton. Silencing of Girdin mRNA results in defective migration in a variety of cultured cells. Moreover, knockout of Girdin causes phenotypes related to defective migration, including hypoplasia of olfactory bulbs and a widened rostral migratory stream (RMS) in mice. To elucidate the molecular basis underlying cellular migration, we generated site- and phosphorylation state-specific antibodies against human Girdin peptides carrying four putative phosphorylation sites (serine1386 [S1386], S1416, tyrosine1764 [Y1764] and Y1798) that had been identified by mutagenesis analyses or mass spectrometric studies. We found that these residues were phosphorylated in an epidermal growth factor (EGF)-dependent manner. Among the four antibodies we developed, the antibody that targeted Girdin when phosphorylated at Y1798 (pY1798) worked well for immunohistochemistry of paraffin-embedded tissues as well as for cultured cells. Immunocytochemistry of HEK293FT cells transfected with an EGF receptor expression plasmid exhibited punctate signals with pY1798. These signals colocalized with those of endocytosed EGF receptors after EGF stimulation. Signals from pY1798 were also observed on lamellipodia, filopodia, focal adhesion and stress fibers in NIH3T3 cells under conventional culture conditions. Immunohistochemistry of paraffin-embedded mouse brain at P14 using anti-pY1798 antibody displayed signals at the hilum-side (internal side) of the dentate gyrus of the hippocampus, the RMS, the accessory olfactory bulb and the olfactory bulb in which Girdin expression was detected. Primary culture of RMS neurons showed punctate signals of pY1798 at the tips of leading processes as well as in the cytoplasm, whereas no signals were observed when neurons were treated with Src inhibitor, PP2. Our data revealed the changes in the phosphorylation status of Y1798 in Girdin when it associated with migration-related structures in vitro and in vivo., (Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.)

A new strategy for the assembly of graphene-like C3N4 on graphene is reported. Transmission electron microscopy (TEM), thermogravimetric analysis (TGA), Raman spectroscopy, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and X-ray photoelectron spectroscopy (XPS) clearly demonstrated that g-C3N4 was successful in the reduction of GO and the immobilization of g-C3N4 on the graphene surface. Investigation of their electrochemical behaviour for several redox systems was conducted, which indicated the unique electron structure endows the G-g-C3N4 with faster transfer and proper amount of g-C3N4 could make G-g-C3N4 advantageous in terms of improving the redox current and promoting electron transfer. Finally, several electroactive biomolecules, such as uric acid (UA), norepinephrine (NE), tyrosine (Tyr), tryptophan (Trp), acetaminophen (APAP) and rutin, were used to probe the biosensing capacity of G-g-C3N4 films by using the cyclic voltammetric method., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Structural changes, L(⁎)-value, cooking yield changes and textural properties of pork frankfurters containing 1% or 2% salt, produced by the two methods were studied by Raman spectroscopy and texture profile analysis. Increasing salt content from 1% to 2% increased the L(⁎)-value, cooking yield and hardness, and decreased (p<0.05) the C-H stretching and CH2 and CH3 bending vibrations, but did not affect the changes of secondary structures, tryptophan or tyrosine residues. Compared with the chopping, the beating increased L(⁎)-value, cooking yield and hardness of the frankfurters in both salt concentrations. It also resulted in an increase in β-sheets, accompanied by a significant (p<0.05) decrease in α-helix content, a greater exposure of tyrosine residues to the polar environment and a decrease in the CH stretching and CH2 and CH3 bending vibrations. The results showed that the beating process enabled lowering of the salt content while improving the L(⁎)-value, cooking yield and hardness of the frankfurters., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Unlabelled: The aim of this study was to evaluate the role of oxidative and nitrosative stress in autogenous bone grafts to the mandible based on immunohistochemical analysis., Material and Methods: Using a well-established sheep model autogenous bone grafts were harvested form the iliac bone. A combination of a Collagen Membrane (CM) and Deproteinized Bovine Bone Material (DBBM) was used to cover the bone graft (Experiment 2). This modification was compared with simple onlay bone grafts (Experiment 1). Immunohistochemically, the expression of specific stable degradation products of oxidative and nitrosative stress was compared between the two experimental groups., Results: Specific markers for oxidative and nitrosative stress showed statistically significant differences in expression in the different experimental groups. The influence of oxidative and nitrosative stress on osteoblasts (OB), osteoclasts (OC), and osteocytes (OCy) was analysed. Experiment 2 showed increased expression of markers in OB and decreased expression in OC., Conclusions: Taking the result of this study and reports from the literature into consideration grafts in Experiment 2 showed less resorption and atrophy, higher activity of OB and inhibition of OC, and less expression of Reactive Oxygen and Nitrogen Species (RONS) as markers of oxidative stress within the graft. These data illustrate the improved remodelling processes in grafts using CM and DBBM., (Copyright © 2013 European Association for Cranio-Maxillo-Facial Surgery. Published by Elsevier Ltd. All rights reserved.)

A novel biosensor based on Fe-doped hydroxyapatite (Fe-HA) nanoparticles and tyrosinase has been developed for the detection of L-tyrosine. Nanostructured Fe-HA was synthesized by a simple microwave irradiation method, and its phase formation, morphology and magnetic property were examined by powder X-ray diffraction (XRD), transmission electron microscopy (TEM) and vibrating sample magnetometer (VSM). Electrochemical performance of the nano Fe-HA/tyrosinase modified glassy carbon electrode (GCE) for detection of L-tyrosine was investigated by cyclic voltammetry (CV) and amperometric methods. The fabricated biosensor exhibited a linear response to L-tyrosine over a wide concentration range of 1.0×10(-7) to 1.0×10(-5) M with a detection limit of 245 nM at pH 7.0. In addition, the fabricated sensor showed an excellent selectivity, good reproducibility, long-term stability and anti-interference towards the determination of L-tyrosine., (© 2013.)

A reliable sensor was fabricated by modifying an acetylene black paste electrode with graphene (denoted as GR/ABPE) for sensitive and selective determination of tryptophan (Trp). Due to the high sorption ability, large surface area and numerous active sites, the GR/ABPE showed a strong enhancement effect on the oxidation of Trp, and greatly increased the peak current. The parameters affecting the Trp determination were investigated. In 1.0 M H2SO4 the voltammetric responses of Trp and tyrosine (Tyr) were well separated into two distinct peaks with peak potential difference (ΔE(pa)) of 115 mV. Under the optimized conditions, in the presence of 0.1 mM Tyr, the oxidation peak current of Trp was proportional to its concentration in the range between 0.1 μM and 0.1 mM, with the limit of detection of 60 nM (S/N=3). The GR/ABPE was applied to the direct detection of Trp in pharmaceutical and biological samples with satisfactory results. This work provides a simple and easy approach to selective detection of Trp in the presence of Tyr., (© 2013.)

Peroxynitrite-mediated protein tyrosine nitration represents a crucial pathogenic mechanism of stroke. Hydroxysafflor yellow A (HSYA) is the most important active component of the safflower plant. Here we assess the neuroprotective efficacy of HSYA and investigate the mechanism through anti-nitrative pathway. Rats were subjected to 60-min ischemia followed by reperfusion. HSYA (2.5-10mg/kg) was injected at 1h after ischemia onset. Other groups received HSYA (10mg/kg) treatment at 3-9h after onset. Infarct volume, brain edema, and neurological score were evaluated at 24h after ischemia. Nitrotyrosine and inducible NO synthase (iNOS) expression, as well as NO level (nitrate/nitrite) in ischemic cortex was examined within 24h after ischemia. The ability of HSYA to scavenge peroxynitrite was evaluated in vitro. Infarct volume was significantly decreased by HSYA (P<0.05), with a therapeutic window of 3h after ischemia at dose of 10mg/kg. HSYA treatment also reduced brain edema and improved neurological score (P<0.05). Nitrotyrosine formation was dose- and time-dependently inhibited by HSYA. The time window of HSYA in decreasing protein tyrosine nitration paralleled its action in infarct volume. HSYA also greatly reduced iNOS expression and NO content at 24h after ischemia, suggesting prevention of peroxynitrite generation from iNOS. In vitro, HSYA blocked authentic peroxynitrite-induced tyrosine nitration in bovine serum albumin and primary cortical neurons. Collectively, our results indicated that post-ischemic HSYA treatment attenuates brain ischemic injury which is at least partially due to reducing nitrotyrosine formation, possibly by the combined mechanism of its peroxynitrite scavenging ability and its reduction in iNOS production., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Methionyl aminopeptidase 2 (MetAP2) plays an important role in the regulation of angiogenesis. This study examined whether nitration of MetAP2 alters its enzymatic activity in vitro. The activity of unmodified, nitrated and oxidised MetAP2 was assessed and it was found that nitration significantly reduced its ability to cleave a chromogenic substrate. Mass spectrometry analysis identified Tyr336 as a nitrated residue in MetAP2. Structural and evolutionary analysis indicate that this is an important residue for MetAP2 activity. Combined, the results show that the activity of MetAP2 is reduced by nitration and raise the possibility that nitration of MetAP2 is a mechanism contributing to endothelial dysfunction., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Testicular damage is a common clinical problem in diabetic individuals that severely affects the quality of life. The present study investigates the possible protective mechanisms of telmisartan, an angiotensin II-receptor antagonist in the germ cell of diabetic rat. Male SD rats were used and randomized into six groups: control, telmisartan control, diabetic control and diabetic group treated with telmisartan at the doses of 3, 6 and 12mg/kg/day, per oral for 4 weeks. Diabetes was induced by injecting a single dose of streptozotocin (STZ), (55mg/kg) dissolved in ice-cold 10mM citrate buffer; pH 4.4 and administered i.p. immediately after preparation to the SD rats. At the end of the study, rats were sacrificed and the levels of nitrite, superoxide, malondialdehyde (MDA), glutathione (reduced and peroxidase) and superoxide dismutase (SOD) were measured. Germ cell toxicity was evaluated by using sperm count, sperm comet assay, histology of testes and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. Further to confirm the oxidative and nitrosative damage, immunohistological quantification of 8-oxo-dG (8-oxo-7,8-dihydro-2'-deoxyguanosine) and 3-nitrotyrosine were evaluated respectively. Results showed that telmisartan significantly restored the levels of nitrite, superoxide, malondialdehyde, and glutathione and superoxide dismutase in diabetic testes. Further, telmisartan significantly increased the sperm counts, reduced apoptotic cell death, sperm DNA damage, oxidative and nitrosative damage in diabetic rat. Western blot analysis showed that telmisartan reduced the testicular inflammation and cell death by down-regulating the expression of NF-κB, IL-6, TNF-α, p-ERK1/2, iNOS, caspase-3 and increasing the PPAR-γ expression. Results clearly indicate that telmisartan significantly reduced the both oxidative and nitrosative stress, inflammation and cell death in diabetic testes. The present results confirmed that telmisartan exhibited beneficial role in the germ cell of diabetic rat., (Copyright © 2013 Elsevier B.V. All rights reserved.)

The composition of three porcine muscles (Longissimus thoracis: LT, Semitendinosus: ST, Masseter: MS) was characterized and its link with muscle quality was evaluated. The LT muscle had a higher content of tyrosine, tryptophan, and carbohydrates and a lower content of vitamin E and haem iron than the MS muscle, while the ST had similar composition to MS but a lower content of haem iron. Large differences between muscles were observed in relative amounts of most of the major fatty acids. The LT muscle had higher saturated fatty acids (SFA) and n-6:n-3 fatty acid ratio, and lower polyunsaturated fatty acids (PUFA), PUFA:SFA ratio, unsaturation index and average fatty acid chain length than the ST and MS muscles. Muscle pH, redness and chroma were positively correlated with vitamin E and unsaturated lipids and negatively correlated with tyrosine, tryptophan, carbohydrates and saturated lipids, whereas muscle lightness and expressible juice showed similar correlations but an opposite sign with these variables., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

An electrochemical sensor based on tricobalt tetroxide nanoparticles-graphene nanocomposite film modified glassy carbon electrodes (GCEs) for sensitive determination of L-tyrosine (L-Tyr) was presented here. The nanoparticles were fabricated by electro-polymerization technology. Scanning electron microscopy was implemented to characterize morphology of the nanocomposite film. The electron transfer behavior of modified electrodes was investigated in 5 mM K3[Fe(CN)6]/K4[Fe(CN)6] solution using cyclic voltammetry and electrochemical impedance spectroscopy (EIS). The electrochemical response of modified electrodes toward L-Tyr was investigated by cyclic voltammetry (CV), square wave voltammetry (SWV), amperometry in detail. The results indicated that synergistic effect of Co3O4 NPs and graphene film dramatically improved the conductivity and sensitivity of the sensor. Under optimal conditions, a wide linear relationship between the responses and L-Tyr concentrations ranging from 1.0×10(-8) to 4.0×10(-5) mol L(-1) was obtained with a comparatively low detection limit of 1.0×10(-9) mol L(-1). Furthermore, the sensor also displays excellent sensitivity and high stability. To further study the practical applicability of the fabricated sensor, it was applied to detect real samples and the received results were satisfactory., (Copyright © 2013 Elsevier B.V. All rights reserved.)

To identify the irradiated meats, various parameters that affect extraction efficiency of tyrosine positional isomers were evaluated. The optimum procedure employed simple extraction by 0.1% formic acid and protein precipitation by acetone. Baseline separation for the extract was carried out on LC-fluorescence detection (FLD) and LC-tandem mass spectrometry (MS/MS). The LC-FLD and LC-MS/MS method had LOD of 1.7-2.1 and 0.3-0.5 ng/mL, respectively, and showed excellent linear correlation over three orders of magnitude, obtained ideal recoveries (78.68-88.90%) and RSD (≤ 8%). The methods were successfully applied in multiple samples. For o- and m-tyrosine, the order of descending trend was: chicken>beef > hairtail > pork and chicken > hairtail > beef > pork, respectively. The radiation dose could be quantitatively evaluated by the nonlinear correlation (y = A(0) x (2) + A(1)x + A(2)) with coefficients of determination r(2) > 0.998 in individual meat samples., (Copyright © 2012 Elsevier Ltd. All rights reserved.)