scholarly journals Ex vivo performance evaluation of a transdermal gold injectable microneedle for tyrosinase sensing

2021 ◽  
Author(s):  
Sorour Darvishi ◽  
Hubert Girault
Keyword(s):  
Redox State ◽  
High Selectivity ◽  
Ex Vivo ◽  
High Sensitivity ◽  
Liquid Electrolyte ◽  
Glucose Detection ◽  
Electrolyte Layer ◽  
Gold Sputtering ◽  
Relative Standard ◽  
Excellent Reproducibility

Here, the semi-invasive direct electrochemical detection of melanoma biomarkers in non-treated skin has been envisaged. The enzyme tyrosinase (TYR) was to be addressed with a microneedle electrochemical sensor. The microneedles were fabricated by polydimethylsiloxane (PDMS) casting with stable polymers. The as-prepared microneedles (MNs) were then coated by gold sputtering. The gold MNs were finally covered by alginate/catechol to provide a liquid electrolyte layer that contained electroactive species, such as catechol, whose redox state can be linked to the concentration of TYR in the skin. The sensor showed high sensitivity of 7.52 µA·mg–1·mL TYR in dummy skin. A relative standard deviation (RSD) of 8.54 % (n=5) demonstrated that the Catechol@alginate: gold MN electrode had excellent reproducibility for continuous glucose detection. Also, five parallel Catechol@alginate: gold MN electrodes were fabricated using the same experimental setup and showed an RSD of 3.95 % (n=5). These results suggest that the electrode fabrication process was reproducible, and Catechol@alginate: gold MN biosensors demonstrated high stability for the repetitive detection of TYR. Furthermore, the sensor has high selectivity in the presence of interfering components towards TYR screening.

Atomic absorption determination of Hg (II), Cd (II), and As (III) in natural and waste water after preliminary group preconcentration

2019 ◽  
Vol 85 (2) ◽  
pp. 12-16
Author(s):  
I. V. Saunina ◽  
E. N. Gribanov ◽  
E. R. Oskotskaya
Keyword(s):  
Waste Water ◽  
Atomic Absorption ◽  
High Sensitivity ◽  
Distribution Coefficients ◽  
Neutral Medium ◽  
Atomic Absorption Determination ◽  
Absorption Determination ◽  
Relative Standard ◽  
Preliminary Group

The sorption of Hg (II), Cd (II), and As (III) by natural aluminosilicate is studied. It is shown that the mineral absorbs those toxicants in a rather wide pH range, quantitative extraction of analytes being achieved in a neutral or close to neutral medium (pH values range within 7.0 - 8.0; 6.3 - 7.5; 7.4 - 8.5 for Hg (II), As (III), and Cd (II), respectively). The effect of the time of phase contact on the degree of extraction of elements is shown. The sorption capacity of the mineral in optimal conditions of the medium acidity (0.06 mmol/g for mercury, 0.31 mmol/g for cadmium, and 0.52 mmol/g for arsenic) is determined. The distribution coefficients attain values of aboutnX 103-nX 104. A new combined method for determination of Hg (II), Cd (II), and As (III) in natural and waste water is developed and tested. The method consists in a preliminary group sorption concentration of the analytes by aluminosilicate, desorption of the analytes from the surface of the mineral and their subsequent atomic absorption determination. The correctness of the method is verified in analysis of spiked samples. The method is easy to use and exhibits high sensitivity, reproducibility and accuracy of analyte determination. The relative standard deviation does not exceed 0.13. Economic availability and possibility of using domestic sorption materials are the important advantages of the proposed procedure which can be used in the practice of laboratories monitoring the quality and safety of environmental objects.

scholarly journals Development of TLC Chromatographic-Densitometric Procedure for Qualitative and Quantitative Analysis of Ceftobiprole

Processes ◽  
2021 ◽  
Vol 9 (4) ◽  
pp. 708
Author(s):  
Żaneta Binert-Kusztal ◽  
Małgorzata Starek ◽  
Joanna Żandarek ◽  
Monika Dąbrowska
Keyword(s):  
High Sensitivity ◽  
Biologically Active ◽  
Fifth Generation ◽  
Ich Guidelines ◽  
Relative Standard ◽  
Lipophilicity Parameter ◽  
Alkaline Environments ◽  
Acetic Acid Water ◽  
Cephalosporin Antibiotics ◽  
The Stability

Currently, there is still a need for broad-spectrum antibiotics. The new cephalosporin antibiotics include, among others, ceftobiprole, a fifth-generation gram-positive cephalosporin, active against Staphylococcus aureus methicillin agonist (MRSA). The main focus of the work was to optimize the conditions of ceftobiprole qualitative determination and to validate the developed procedure according to ICH guidelines. As a result of the optimization process, HPTLC Cellulose chromatographic plates as a stationary phase and a mixture consisting of ethanol:2-propanol: glacial acetic acid: water (4:4:1:3, v/v/v/v) as a mobile phase were chosen. The densitometric detection was carried out at maximum absorbance of ceftobiprole (λ = 232 nm). Next, the validation process of the developed procedure was carried out. The relative standard deviation (RSD) for precision was less than 1.65%, which proves the high compatibility of the results, as well as the LOD = 0.0257 µg/spot and LOQ = 0.0779 µg/spot values, which also confirm the high sensitivity of the procedure. The usefulness of the developed method for the stability studies of ceftobiprole was analyzed. Study was carried out under stress conditions, i.e., acid and alkaline environments, exposure to radiation imitating sunlight and high temperature (40–60 °C). It was found that cefotbiprole is unstable in an alkaline environment and during exposure to UV-VIS radiation. Moreover, the lipophilicity parameter, as a main physicochemical property of the biologically active compound, was determined using experimental and computational methods.

Phytic acid functionalized magnetic bimetallic metal–organic frameworks for phosphopeptide enrichment

2021 ◽  
Vol 9 (7) ◽  
pp. 1811-1820
Author(s):  
Shuang Yan ◽  
Bin Luo ◽  
Jia He ◽  
Fang Lan ◽  
Yao Wu
Keyword(s):  
Phytic Acid ◽  
Efficient Method ◽  
High Selectivity ◽  
Metal Organic Framework ◽  
Metal Organic Frameworks ◽  
High Sensitivity ◽  
Phosphopeptide Enrichment ◽  
Metal Organic ◽  
Organic Framework

Novel bimetallic metal–organic framework nanocomposites were fabricated by a facile yet efficient method. The as-prepared nanomaterial exhibited high sensitivity and high selectivity toward phosphopeptides and good reusability of five cycles for enriching phosphopeptides.

scholarly journals Engineered Phage Endolysin Eliminates Gardnerella Biofilm without Damaging Beneficial Bacteria in Bacterial Vaginosis Ex Vivo

Pathogens ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 54
Author(s):  
Christine Landlinger ◽  
Lenka Tisakova ◽  
Vera Oberbauer ◽  
Timo Schwebs ◽  
Abbas Muhammad ◽  
...  
Keyword(s):  
Bacterial Vaginosis ◽  
Bactericidal Activity ◽  
High Selectivity ◽  
Ex Vivo ◽  
Vaginal Microbiome ◽  
Wild Type ◽  
Wild Type Enzyme ◽  
Promising Alternative ◽  
First Time

Bacterial vaginosis is characterized by an imbalance of the vaginal microbiome and a characteristic biofilm formed on the vaginal epithelium, which is initiated and dominated by Gardnerella bacteria, and is frequently refractory to antibiotic treatment. We investigated endolysins of the type 1,4-beta-N-acetylmuramidase encoded on Gardnerella prophages as an alternative treatment. When recombinantly expressed, these proteins demonstrated strong bactericidal activity against four different Gardnerella species. By domain shuffling, we generated several engineered endolysins with 10-fold higher bactericidal activity than any wild-type enzyme. When tested against a panel of 20 Gardnerella strains, the most active endolysin, called PM-477, showed minimum inhibitory concentrations of 0.13–8 µg/mL. PM-477 had no effect on beneficial lactobacilli or other species of vaginal bacteria. Furthermore, the efficacy of PM-477 was tested by fluorescence in situ hybridization on vaginal samples of fifteen patients with either first time or recurring bacterial vaginosis. In thirteen cases, PM-477 killed the Gardnerella bacteria and physically dissolved the biofilms without affecting the remaining vaginal microbiome. The high selectivity and effectiveness in eliminating Gardnerella, both in cultures of isolated strains as well as in clinically derived samples of natural polymicrobial biofilms, makes PM-477 a promising alternative to antibiotics for the treatment of bacterial vaginosis, especially in patients with frequent recurrence.

scholarly journals A novel method for visualizing and tracking endogenous mRNA in a specific cell population in pathological neovascularization

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Md Imam Uddin ◽  
Tyler C. Kilburn ◽  
Sara Z. Jamal ◽  
Craig L. Duvall ◽  
John S. Penn
Keyword(s):  
Real Time ◽  
Disease Burden ◽  
Ex Vivo ◽  
Expression Patterns ◽  
Fluorescence Emission ◽  
High Sensitivity ◽  
Living Systems ◽  
Specific Cell ◽  
Potential Marker

AbstractDiabetic retinopathy, retinopathy of prematurity and retinal vein occlusion are potentially blinding conditions largely due to their respective neovascular components. The development of real-time in vivo molecular imaging methods, to assess levels of retinal neovascularization (NV), would greatly benefit patients afflicted with these conditions. mRNA hybridization techniques offer a potential method to image retinal NV. The success of these techniques hinges on the selection of a target mRNA whose tissue levels and spatial expression patterns correlate closely with disease burden. Using a model of oxygen-induced retinopathy (OIR), we previously observed dramatic increases in retinal endoglin that localized to neovascular structures (NV), directly correlating with levels of neovascular pathology. Based on these findings, we have investigated Endoglin mRNA as a potential marker for imaging retinal NV in OIR mice. Also of critical importance, is the application of innovative technologies capable of detecting mRNAs in living systems with high sensitivity and specificity. To detect and visualize endoglin mRNA in OIR mice, we have designed and synthesized a novel imaging probe composed of short-hairpin anti-sense (AS) endoglin RNA coupled to a fluorophore and black hole quencher (AS-Eng shRNA). This assembly allows highly sensitive fluorescence emission upon hybridization of the AS-Eng shRNA to cellular endoglin mRNA. The AS-Eng shRNA is further conjugated to a diacyl-lipid (AS-Eng shRNA–lipid referred to as probe). The lipid moiety binds to serum albumin facilitating enhanced systemic circulation of the probe. OIR mice received intraperitoneal injections of AS-Eng shRNA–lipid. Ex vivo imaging of their retinas revealed specific endoglin mRNA dependent fluorescence superimposed on neovascular structures. Room air mice receiving AS-Eng shRNA–lipid and OIR mice receiving a non-sense control probe showed little fluorescence activity. In addition, we found that cells in neovascular lesions labelled with endoglin mRNA dependent fluorescence, co-labelled with the macrophage/microglia-associated marker IBA1. Others have shown that cells expressing macrophage/microglia markers associate with retinal neovascular structures in proportion to disease burden. Hence we propose that our probe may be used to image and to estimate the levels of retinal neovascular disease in real-time in living systems.

scholarly journals Serum Albumin Redox States: More than Oxidative Stress Biomarker

Antioxidants ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 503
Author(s):  
Fuka Tabata ◽  
Yasuaki Wada ◽  
Satomi Kawakami ◽  
Kazuhiro Miyaji
Keyword(s):  
Oxidative Stress ◽  
Serum Albumin ◽  
Animal Studies ◽  
Redox State ◽  
Ex Vivo ◽  
Pathological Condition ◽  
Analytical Techniques ◽  
Cysteine Residue ◽  
Research Field ◽  
Systemic Oxidative Stress

Serum albumin is the most abundant circulating protein in mammals including humans. It has three isoforms according to the redox state of the free cysteine residue at position 34, named as mercaptalbumin (reduced albumin), non-mercaptalbumin-1 and -2 (oxidized albumin), respectively. The serum albumin redox state has long been viewed as a biomarker of systemic oxidative stress, as the redox state shifts to a more oxidized state in response to the severity of the pathological condition in various diseases such as liver diseases and renal failures. However, recent ex vivo studies revealed oxidized albumin per se could aggravate the pathological conditions. Furthermore, the possibility of the serum albumin redox state as a sensitive protein nutrition biomarker has also been demonstrated in a series of animal studies. A paradigm shift is thus ongoing in the research field of the serum albumin. This article provides an updated overview of analytical techniques for serum albumin redox state and its association with human health, focusing on recent findings.

Ex Vivo assessment of competent strut coverage after coronary stenting by optical coherence tomography

2020 ◽  
Vol 41 (Supplement_2) ◽  
Author(s):  
R Bhoite ◽  
H Jinnouchi ◽  
F Otsuka ◽  
Y Sato ◽  
A Sakamoto ◽  
...  
Keyword(s):  
Optical Coherence Tomography ◽  
Predictive Value ◽  
Ex Vivo ◽  
High Sensitivity ◽  
Optical Coherence ◽  
Cross Sectional ◽  
Neointimal Thickness ◽  
Stent Strut ◽  
Oct Imaging ◽  
Definition Of

Abstract Background In many studies, struts coverage is defined as >0 mm of tissue overlying the stent struts by optical coherence tomography (OCT). However, this definition has never been validated using histology as the “gold standard”. The present study sought to assess the appropriate cut-off value of neointimal thickness of stent strut coverage by OCT using histology. Methods OCT imaging was performed on 39 human coronary arteries with stents from 25 patients at autopsy. A total of 165 cross-sectional images from 46 stents were co-registered with histology. The optimal cut-off value of strut coverage by OCT was determined. Strut coverage by histology was defined as endothelial cells with at least underlying two layers of smooth muscle cells. Considering the resolution of OCT is 10–20 μm, 3 different cut-off values (i.e. at ≥20, ≥40, and ≥60 μm) were assessed. Results A total of 2235 struts were evaluated by histology. Eventually, 1216 struts which were well-matched struts were analyzed in this study. By histology, uncovered struts were observed in 160 struts and covered struts were observed in 1056 struts. The broadly used definition of OCT-coverage which does not consider neointimal thickness yielded a poor specificity of 37.5% and high sensitivity 100%. Of 3 cut-off values, the cut-off value of >40 μm was more accurate as compared to >20 and >60 mm [sensitivity (99.3%), specificity (91.0%), positive predictive value (98.6%), and negative predictive value (95.6%)] Conclusion The most accurate cut-off value was ≥40 μm neointimal thickness by OCT in order to identify stent strut coverage validated by histology. Funding Acknowledgement Type of funding source: None

Core-shell Cu@C@ZIF-8 composite: a high-performance electrode material for electrochemical sensing of nitrite with high selectivity and sensitivity

2021 ◽  
Author(s):  
Feng Gao ◽  
Xiaolong Tu ◽  
Yongfang Yu ◽  
Yansha Gao ◽  
Jin Zou ◽  
...  
Keyword(s):  
High Performance ◽  
High Selectivity ◽  
Limit Of Detection ◽  
Metal Organic Framework ◽  
High Sensitivity ◽  
Electrochemical Sensing ◽  
Core Shell ◽  
Zeolitic Imidazolate Framework ◽  
Sensing Platform ◽  
The Difference

Abstract Herein, an efficient electrochemical sensing platform is proposed for selective and sensitive detection of nitrite on the basis of Cu@C@Zeolitic imidazolate framework-8 (Cu@C@ZIF-8) heterostructure. Core-shell Cu@C@ZIF-8 composite was synthesized by pyrolysis of Cu-metal-organic framework@ZIF-8 (Cu-MOF@ZIF-8) in Ar atmosphere on account of the difference of thermal stability between Cu-MOF and ZIF-8. For the sensing system of Cu@C@ZIF-8, ZIF-8 with proper pore size allows nitrite diffuse through the shell, while big molecules cannot, which ensures high selectivity of the sensor. On the other hand, Cu@C as electrocatalyst promotes the oxidation of nitrite, thereby resulting high sensitivity of the sensor. Accordingly, the Cu@C@ZIF-8 based sensor presents excellent performance for nitrite detection, which achieves a wide linear response range of 0.1 µM to 300.0 µM, and a low limit of detection (LOD) of 0.033 µM. In addition, the Cu@C@ZIF-8 sensor possesses excellent stability and reproducibility, and was employed to quantify nitrite in sausage samples with recoveries of 95.45-104.80%.

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