Comparative Analysis of Catalase Activity in Plants: Spectrophotometry and Native PAGE Approaches.

Catalase, a pivotal enzyme in plant antioxidative defense mechanisms, plays a crucial role in detoxifying hydrogen peroxide, a reactive oxygen species (ROS). In this chapter, a comparative analysis of catalase activity was conducted using two distinct methodologies: spectrophotometry and non-denaturing polyacrylamide gel electrophoresis (PAGE). The spectrophotometric approach allowed the quantification of catalase activity by measuring the breakdown rate of hydrogen peroxide, while native PAGE enabled the separation and visualization of catalase isozymes, based on their native molecular weight and charge characteristics, and specific staining assay. Both methods provide valuable insights into catalase activity, offering complementary information on the enzyme's functional diversity and distribution within different plant tissues. This study integrates different techniques, previously described, to comprehensively elucidate the role of catalase in plant metabolism. Furthermore, it provides the possibility of obtaining a holistic understanding of antioxidant defense mechanisms by considering both total activity and isoenzyme distribution of catalase enzyme.

Change in Osmotic Pressure Influences the Absorption Spectrum of Hemoglobin inside Red Blood Cells.

Absorption spectra of red blood cell (RBC) suspensions are investigated in an osmolarity range in the medium from 200 mOsm to 900 mOsm. Three spectral parameters are used to characterize the process of swelling or shrinkage of RBC-the absorbance at 700 nm, the Soret peak height relative to the spectrum background, and the Soret peak wavelength. We show that with an increase in the osmolarity, the absorbance at 700 nm increases and the Soret peak relative height decreases. These changes are related to the changes in the RBC volume and the resulting increase in the hemoglobin intracellular concentration and index of refraction. Confocal microscopy and flow cytometry measurements supported these conclusions. The maximum wavelength of the Soret peak increases with increasing osmolarity due to changes in the oxygenation state of hemoglobin. Using these spectrum parameters, the process of osmosis in RBCs can be followed in real time, but it can also be applied to various processes, leading to changes in the volume and shape of RBCs. Therefore, we conclude that UV-Vis absorption spectrophotometry offers a convenient, easily accessible, and cost-effective method to monitor changes in RBC, which can find applications in the field of drug discovery and diagnostics of RBC and hemoglobin disorders.

Development of a point-of-care colorimetric metabolomic sensor platform.

Metabolomics is the large-scale study of small molecule metabolites within a biological system. It has applications in measuring dietary intake, predicting heart disease risk, and diagnosing cancer. Metabolites are often measured using high-end analytical tools such as mass spectrometers or large spectrophotometers. However, due to their size, cost, and need for skilled operators, using such equipment at the bedside is not practical. To address this issue, we have developed a low-cost, portable, optical color sensor platform for metabolite detection. This platform includes LEDs, sensors, microcontrollers, a power source, and a Bluetooth chip enclosed within a 3D-printed light-tight case. We evaluated the color sensor's performance using both a range of dyed water samples as well as well-established colorimetric reactions for specific metabolite detection. The sensor accurately measured creatinine, L-carnitine, ascorbate, and succinate well within normal human urine levels with accuracy and sensitivity equal to or better than a standard laboratory spectrophotometer. Our color sensor offers a cost-effective, portable alternative for measuring metabolites via colorimetric assays, thereby enabling low-cost, point-of-care metabolite testing.

Color matching and color recovery in large composite restorations using single-shade or universal composites.

This study assessed the color-matching ability and color recovery of unprepared teeth when using single-shade composites and a universal composite in large restorations. Buccal and palatine surface colors of molars were measured with a spectrophotometer (CIELAB) before preparing round cavities (6 mm in diameter, 2 mm in depth). The cavities were randomly filled with a single-shade composite (Omnichroma, Diamond One, or Vittra APS Unique) or a universal composite (Filtek Universal). Color measurements of the restored cavities were taken, and overall color differences (ΔEab and ΔE00) and differences in the whitening index for dentistry (ΔWID) from baseline were calculated. Additionally, visual assessments of a color match to the surrounding enamel were performed by forty evaluators (laypersons and undergraduate students of dentistry) in a viewing booth under illuminant D65, with rating scores from 0 (no color mismatch) to 4 (not acceptable). Data were analyzed using RM or one-way ANOVA (α = 0.05). Results showed that the restorations generally exhibited whiter colors (WID ranged from 27.9 to 41.3) than the unprepared teeth (WID ranged from 15.9 to 19.3). The composite Filtek Universal demonstrated the lowest color discrepancy (ΔWID = 8.6; ΔE00 = 10.8; and ΔE00 = 6.2), and no significant differences were observed among the evaluated single-shade composites. Furthermore, all composites showed similar and adequate color matches to the surrounding enamel. However, it is important to note that despite their ability to match the surrounding enamel reasonably, none of the composites evaluated in large restorations fully recovered the color observed in unprepared teeth.

Investigation on the Validity of in vitro Sun Protection Factor and Ultraviolet-A Protection Factor Evaluation Method for Sunscreen Samples.

Factors affecting the in vitro Sun Protection Factor (SPF) and Ultraviolet-A Protection Factor (UVA-PF) of sunscreens were analyzed for verifying the validity and reliability of the ISO24443 evaluation method. UV absorbance measurements by different spectrophotometers did not lead to the large difference in in vitro SPF/UVA-PF, although the UV absorbance determined by each spectrophotometer exhibited relatively large difference when it was larger than 2. On the other hand, relatively large difference was found in in vitro SPF/UVA-PF by utilizing European Cosmetic and Perfumery Association (Colipa) 1994 or UV-solar simulated radiation (UV-SSR) for the spectral irradiance. Appropriateness of employing the coefficient of adjustment for the determination of in vitro UVA-PF was also found to be reexamined.

Spider chart, greenness and whiteness assessment of experimentally designed multivariate models for simultaneous determination of three drugs used as a combinatory antibiotic regimen in critical care units: Comparative study.

In this study, five earth-friendly spectrophotometric methods using multivariate techniques were developed to analyze levofloxacin, linezolid, and meropenem, which are utilized in critical care units as combination therapies. These techniques were used to determine the mentioned medications in laboratory-prepared mixtures, pharmaceutical products and spiked human plasma that had not been separated before handling. These methods were named classical least squares (CLS), principal component regression (PCR), partial least squares (PLS), genetic algorithm partial least squares (GA-PLS), and artificial neural network (ANN). The methods used a five-level, three-factor experimental design to make different concentrations of the antibiotics mentioned (based on how much of them are found in the plasma of critical care patients and their linearity ranges). The approaches used for levofloxacin, linezolid, and meropenem were in the ranges of 3-15, 8-20, and 5-25 µg/mL, respectively. Several analytical tools were used to test the proposed methods' performance. These included the root mean square error of prediction, the root mean square error of cross-validation, percentage recoveries, standard deviations, and correlation coefficients. The outcome was highly satisfactory. The study found that the root mean square errors of prediction for levofloxacin were 0.090, 0.079, 0.065, 0.027, and 0.001 for the CLS, PCR, PLS, GA-PLS, and ANN models, respectively. The corresponding values for linezolid were 0.127, 0.122, 0.108, 0.05, and 0.114, respectively. For meropenem, the values were 0.230, 0.222, 0.179, 0.097, and 0.099 for the same models, respectively. These results indicate that the developed models were highly accurate and precise. This study compared the efficiency of artificial neural networks and classical chemometric models in enhancing spectral data selectivity for quickly identifying three antimicrobials. The results from these five models were subjected to statistical analysis and compared with each other and with the previously published ones. Finally, the whiteness of the methods was assessed by the recently published white analytical chemistry (WAC) RGB 12, and the greenness of the proposed methods was assessed using AGREE, GAPI, NEMI, Raynie and Driver, and eco-scale, which showed that the suggested approaches had the least negative environmental impact. Furthermore, to demonstrate solvent sustainability, a greenness index using a spider chart methodology was employed.

Method development and validation for the quantitative determination of total flavonoids through the complexation of iron (III) and its application in real sample.

BACKGROUND: The determination of flavonoids in real sample using UV-Vis spectrophotometer commonly uses quercetin and catechin with Al+3 complexing agent as reference materials for the calibration of the instrument. However, getting these standard materials is challenging due to its expense and unavailability in the chemical reserve of the country. Moreover, the Al+3 - quercetin complexation standard method demands high amount of quercetin in spite of its high cost. Hence, developing alternative method that can solve this problem is crucial for the determination of flavonoids in the real sample. RESULTS: An iron-based complexation method for the determination of flavonoids in the real sample was developed that reduces the amount of quercetin by 200 times (1 mg/mL to 0.005 mg/mL) during the calibration of UV-Vis spectroscopy as an alternative method. The reaction parameters (incubation time, pH, and concentration of quercetin) were optimized using software Design Expert 11 and confirmed by the practical experiments. The kinetics of reaction between iron and quercetin was found to be pseudo first order with rate constant of kobs at 340 and 510 nm. The analysis window for the flavonoid complex was achieved with the kinetic discrimination of the interferences at its optimized time of complexation 20 min and absorbance maxima of 510 nm. The developed method was validated by evaluating its precision, accuracy, recovery test (84-117%), detection limit and quantification limit following the standard protocols. The calibration of the instrument has been developed for the new method and the linear regression coefficient (R2) of 0.998 was obtained. SIGNIFICANCE: Applying the developed standard material (Fe3+ - quercetin complex) gives freedom for the analytical chemists to find the standard materials that is accessible and cheaper than the existing one (Al3+-quercetin complex). The developed method can also be easily applied for determination of flavonoid in the real samples without potential interferences coming from sample matrix.

Development of Transmission Ambient Pressure Laser Desorption Ionization/Postphotoionization Mass Spectrometry Imaging.

Laser-based high-resolution mass spectrometry imaging at ambient conditions has promising applications in life science. However, the ion yield during laser desorption/ablation is poor. Here, transmission atmospheric pressure laser desorption ionization combined with a compact postphotoionization (t-AP-LDI/PI) assembly with a krypton discharge lamp was developed for the untargeted imaging of various biomolecules. The spatial distributions of numerous lipid classes, fatty acids, neurotransmitters, and amino acids in the subregions of mouse cerebellum tissue were obtained. Compared with single laser ablation, the sensitivities for most analytes were increased by 1 to 3 orders of magnitude by dopant-assisted postphotoionization. After careful optimization, a spatial resolution of 4 µm could be achieved for the metabolites in mouse hippocampus tissue. Finally, the melanoma tissue slices were analyzed using t-AP-LDI/PI MSI, which revealed the metabolic heterogeneity of the melanoma microenvironment and exhibited the phenomenon of abnormal proliferation and invasion trends in tumor cells.

Influence of repositioning guides' color and usage on precision in tooth color measurement with a clinical spectrophotometer.

BACKGROUND: Repositioning guides are commonly employed in clinical studies to ensure consistent tooth color measurements. Yet, their influence on measured color remains uncertain. OBJECTIVE: This study evaluated the impact of repositioning guides' color and usage on tooth color measurement using a clinical spectrophotometer. METHODOLOGY: In total, 18 volunteers participated in this study, in which the color of their upper left central incisor and upper left canine was measured with or without repositioning guides (control). The guides were made from pink, blue, or translucent silicone, as well as an acetate-based bleaching tray. Tooth color was measured in triplicates using a clinical spectrophotometer based on the CIELAB system. The standard deviations of these readings were used to estimate reproducibility, and color differences (ΔE00) between the measurements with guides and the control were calculated. RESULTS: Repositioning guides had a minimal effect on L* values and no effect on b* values. The use of pink silicone increased a* values, whereas blue or translucent silicone reduced them. Irrespective of the evaluated tooth, the lowest ΔE00 values were observed for the translucent silicone and bleaching tray. The usage of guides only affected data variability for the L* color coordinate. CONCLUSION: Using repositioning guides can significantly impact the precision of tooth color measurement with a clinical spectrophotometer.

Measuring differential fitness costs and interactions between genetic cassettes using fluorescent spectrophotometry.

In this article, we present a method for designing, executing, and analyzing data from a microbial competition experiment. We use fluorescent reporters to label different competing strains and resolve individual growth curves using a fluorescent spectrophotometer. Our comprehensive data analysis pipeline integrates multiple experiments to simultaneously infer sources of variation, extract selection coefficients, and estimate the genetic contributions to fitness for various synthetic genetic cassettes (SGCs). To demonstrate the method, we employ a synthetic biological system based on Escherichia coli. Strains carry 1 of 10 different plasmids and one of three genomically integrated fluorescent markers. All strains are co-cultured to obtain real-time measurements of optical density (total population density) and fluorescence (sub-population densities). We identify challenges in calibrating between fluorescence and density and of fluorescent proteins maturing at different rates. To resolve these issues, we compare two methods of fluorescence calibration and correct for maturation by measuring in vivo maturation times. We provide evidence of genetic interactions occurring between our SGCs and further show how to use our statistical model to test some hypotheses about microbial growth and the costs of protein expression.IMPORTANCEFluorescently labeled co-cultures are becoming increasingly popular. The approach proposed here offers a high standard for experimental design and data analysis to measure selection coefficients and growth rates in competition. Measuring competitive differences is useful in many laboratory studies, allowing for fitness cost-correction of growth rates and ecological interactions and testing hypotheses in synthetic biology. Using time-resolved growth curves, rather than endpoint measurements, for competition assays allows us to construct a detailed scientific model that can be used to ask questions about fine-grained phenomena, such as bacterial growth dynamics, as well as higher-level phenomena, such as the interactions between synthetic cassette expression.

Harmonizing color measurements in dentistry using translucent tooth-colored materials.

BACKGROUND: Since color measurements are relative, the discrepancy among different instruments is alarmingly high. This multicenter study evaluated the effectiveness of instrument calibration and inter-instrument harmonization of different spectrophotometers with the same optical geometry using tooth-colored, translucent dental materials. METHODS: The coordinating center (CC) spectrophotometer was calibrated using the NPL Ceram Series II set. Two sets of 10 specimens, labeled 1 to 10 and I to X (10 mm in diameter and 1 mm thick), were tested at CC and three research sites (RS1, RS2, and RS3) using the same d/8° optical geometry spectrophotometers. Calibration factors were calculated for each material and site to obtain the average calibration factors for sets 1-10, set I-X, and the combination of both. The differences among the non-corrected and corrected reflection values were calculated using CIEDE2000 (DeltaE00) and CIELAB (DeltaEab) color difference formulas and were submitted to ANOVA and Tukey test (α = 0.05). RESULTS: A significant decrease of color differences between non-corrected as compared to corrected measurements was recorded for all CC-RS and RS-RS comparisons. The reduction of DeltaE00 values between non-corrected and corrected for CC-RS1, CC-RS2, and CC-RS3 were 83.1%, 77.2%, and 73.6%, respectively. The corresponding DeltaE00 values for RS1-RS2, RS1-RS3, and RS2-RS3 comparisons, indirectly compared in the experiment, were 84.2%, 82.8%, and 68.5%, respectively. There was a significant reduction of DeltaE00 and DeltaEab color difference for all combined RS pairs and each of three RS pairs, corrected with one of two specimen sets calibration factors separately. CONCLUSIONS: Calibration and harmonization of color measurements in dentistry using tooth-colored, translucent restorative materials significantly decreased measurement discrepancies between the coordinating center and research sites and among pairs of research sites.

Charge transfer complex-based spectrophotometric analysis of famotidine in pure and pharmaceutical dosage forms.

A straightforward and efficient spectrum technique was created using Ortho-chloranil as the electron acceptor (-acceptor) in a charge transfer (CT) complex formation reaction to determine the concentration of famotidine (FMD) in solutions. Compared to the double-distilled blank solution, the reaction result detected a definite violet colour at a maximum absorption wavelength of 546 nm, For concentrations range 2-28 µg/ml, the technique demonstrated excellent compliance with Beer-Law and Lambert's, as evidenced by its molar absorptivity of 2159.648 L mol-1 cm-1. Lower detection limits of 0.3024 µg/ml and 1.471 µg/ml, respectively, were discovered. The complexes of famotidine and Ortho-chloranil were found to have a 2:1 stoichiometry. Additionally, the suggested approach effectively estimated famotidine concentrations in pharmaceutical formulations, particularly in tablet form.

DNA Interaction with Coordination Compounds of Cd(II)containing 1,10-Phenanthroline.

The experimental study of the DNA interaction with three cadmium coordination compounds [Cd(phen)3](CH3CO2)2, [Cd(phen)2(H2O)2](CH3CO2)2, and [Cd2(phen)4(H2O)2](CH3CO2)4 was carried out using spectrophotometry, viscosity, and dynamic light scattering methods. The role of the solution ionic strength (concentration of NaCl) was analyzed. All compounds can penetrate (fully or partly) to the major or minor DNA grooves. It was shown that, in addition to the important role of electrostatic interactions in the formation of the complex, intercalation of the 1,10-phenanthroline ligand occurs for compounds [Cd(phen)2(H2O)2](CH3CO2)2 and [Cd2(phen)4(H2O)2](CH3CO2)4. Compound [Cd(phen)3](CH3CO2)2 binds to DNA externally. The coordination bond between cadmium and DNA was formed in DNA complexes with [Cd2(phen)4(H2O)2](CH3CO2)4. Preliminary computer modeling of the DNA interaction with the compounds used was performed.

Influence of Thickness on the Translucency Parameter and Whiteness Index of Single-Shade Resin Composites.

The study aimed to evaluate the effect of thickness on the translucency parameter (TP) and whiteness index for dentistry (WID) of single-shade resin composites compared with a multi-shade resin composite after thermocycling. A total of 100 resin composite samples of five brands (Filtek Z250, Omnichroma, Vittra APS Unique, Zenchroma, and Charisma Diamond One) with thicknesses of 2 mm and 4 mm were prepared (n=10). CIE L*a*b* coordinates were obtained using a spectrophotometer. The resin composite samples were subjected to 10,000 thermocycles. Before and after thermocycling, TP values were calculated with the CIEDE2000 formula, and whiteness values were calculated using the WID. The influence of thickness, resin composite, and aging on these variables was analyzed using the generalized linear model (p<0.05). Omnichroma and Filtek Z250 exhibited, respectively, the highest and the lowest TP00 values compared with the other resin materials (p<0.001). The TP00 values of Zenchroma and Diamond One were similar (p>0.05). Vittra APS Unique had a lower TP than Zenchroma and Diamond One (p<0.001). Before thermocycling, the highest WID showed the combination of Vittra APS Unique with thicknesses of 2 mm (48.33±1.18). After thermocycling, the highest WID was shown in the combination of Vittra APS Unique with thicknesses of 2 mm (45.33±0.55) and 4 mm (46.23±0.94) (p<0.001). After thermocycling, the lowest WID was for the combination of Filtek Z250 with thicknesses 2 mm (21.16±0.93) and 4 mm (20.70±0.91) (p<0.001). The TP and WID values of the resin composites varied across different composites and thicknesses. Single-shade resin composites of different thicknesses demonstrated higher translucency and whiteness values than multi-shade resin composite, both before and after thermocycling.

Machine Learning-based Deep Analysis of Human Blood using NIR Spectrophotometry Signatures.

BACKGROUND: Non-invasive bio-diagnostics are essential for providing patients with safer treatment. In this subject, significant growth is attained for noninvasive anaemia detection in terms of Hb concentration by means of spectroscopic and image analysis. The lower satisfaction rate is found due to inconsistent results in various patient settings. OBJECTIVE: This observational study aims to present an adaptable point-of-care Near-Infrared (NIR) spectrophotometric approach with a constructive Machine Learning (ML) algorithm for monitoring Haemoglobin (Hb) concentration by considering dominating influencing factors into account. METHODS: To accomplish this objective, 121 subjects (19.2-55.4 years) were enrolled in the study, having a wide range of Hb concentrations (8.2-17.4 g/dL) obtained from two standard Laboratory analyzers. To inspect the performance, the unique dimensionality reduction approaches are applied with numerous regression models using 5-fold cross-validation. RESULTS: The optimum accuracy is found using support vector regression (SVR) and mutual information having 3 independent features i.e. Pearson correlation (r)= 0.79, standard deviation (SD)= 1.07 g/dL, bias=-0.13 g/dL and limits of agreement (LoA)=-2.22 to 1.97 g/dL. Additionally, comparability between two standard laboratory analyzers is found as; r=0.97, SD=0.50 g/dL, bias=0.21 g/dL, and LoA= -0.77 to 1.19 g/dL. CONCLUSION: The precision of ±1 g/dL in 5-fold cross-validation ensures the same performance irrespective of different age groups, gender, BMI, smoking level, drinking level, and skin type. The outcomes with the offered NIR sensing system and an exclusive ML algorithm can accelerate its' requirement at remote locative rural areas and critical care units where continuous Hb monitoring is compulsory.

Simultaneous determination of meloxicam and bupivacaine via a novel modified dual wavelength method and an advanced chemometric approach.

This study presents two spectrophotometric methods; a novel dual wavelength-derivative spectrophotometry and multivariate curve resolution-alternating least squares (MCR-ALS) for the simultaneous determination of a fixed dose combination of bupivacaine (BUP) and meloxicam (MEL) in a ratio of 30:1. The extended UV spectrum of MEL enables its direct determination at λmax 360 nm with no interference from BUP. The determination of BUP was unfeasible directly because the UV spectra of both drugs are moderately overlapped over the wavelength range of 250-450 nm, thus new chemometric based spectrophotometric methods should be developed for its determination. Dual wavelength-derivative method was employed based on using first derivative spectra. The selected dual wavelengths for determination BUP were 274.6 nm and 374.6 nm where the dA/dλ amplitudes differences for MET are equal to zero. MCR-ALS is advanced chemometric tool that enables analysis of multicomponent samples in complex matrices with high resolution based on the decomposition of signal/spectral data into the pure spectra and corresponding concentration profile. The figures of merits for MCR model show that there is a good agreement between the actual and predicted concentrations for MEL and BUP. The methods were validated and statistically compared with a reported HPLC method.

Rapid inverse radiative transfer solver for multiparameter spectrophotometry without integrating sphere.

Significance: Multiparameter spectrophotometry (MPS) provides a powerful tool for accurate characterization of turbid materials in applications such as analysis of material compositions, assay of biological tissues for clinical diagnosis and food safety monitoring. Aim: This work is aimed at development and validation of a rapid inverse solver based on a particle swarm optimization (PSO) algorithm to retrieve the radiative transfer (RT) parameters of absorption coefficient, scattering coefficient and anisotropy factor of a turbid sample. Approach: Monte Carlo (MC) simulations were performed to obtain calculated signals for comparison to the measured ones of diffuse reflectance, diffuse transmittance and forward transmittance. An objective function has been derived and combined with the PSO algorithm to iterate MC simulations for MPS. Results: We have shown that the objective function can significantly reduce the variance in calculated signals by local averaging of an inverse squared error sum function between measured and calculated signals in RT parameter space. For validation of the new objective function for PSO based inverse solver, the RT parameters of 20% Intralipid solutions have been determined from 520 to 1000 nm which took about 2.7 minutes on average to complete signal measurement and inverse calculation per wavelength. Conclusion: The rapid solver enables MPS to be translated into easy-to-use and cost-effective instruments without integrating sphere for material characterization by separating and revealing compositional profiles at the molecular and particulate scales.

Dopamine-based selective spectrophotometry p-aminosalicylic acid assay by hydrolyzate-triggered formation of azamonardine-like products.

BACKGROUND: The selective recognition of drugs and its metabolism or decomposition products is significant to drug development and drug resistance research. Fluorescence-based techniques provide satisfying sensitivity by target-triggered chemical reaction. However, the interference from the matrix or additives usually restricts the specific detection. It is highly desirable to explore specific chemical reactions for achieving selective perception of these species. RESULTS: We report a specific m-aminophenol (MAP)-dopamine (DA) reaction, which generates highly fluorescent azamonardine-like products. Based on this reaction, fluorometric and indirect detection of p-aminosalicylic acid (typical antituberculosis drug, PAS) can be realized using the DA-based probe with high sensitivity. The acid induces the decarboxylation of PAS and produces MAP, which reacts with DA and generates fluorescent azamonardine-like products. The practical application of the proposed method is validated by the accurate PAS analysis in urine samples and Pasinazid tablets. Interestingly, none of additives in the Pasinazid tablets contribute comparable fluorescence variation. SIGNIFICANCE: This work discovers a new MAP-DA reaction for the first time, it not only explores sensitive PAS drug detection probe, but also demonstrates the feasibility of the development of novel drug analysis platform by recognizing decomposition product with specific reaction. Thus, new avenues for the exploration of simple and rapid spectrophotometric probes toward various drug analytes with high specify and sensitivity based on this tactic might be possible in analytical and drug-related fields.

Development of a fluorometric and colorimetric dual-mode sensing platform for acid phosphatase assay based on Fe3+ functionalized CuInS2/ZnS quantum dots.

BACKGROUND: The spectral dual-mode response towards analyte has been attracted much attention, benefiting from the higher detection accuracy of such strategy in comparison to single signal readout. However, the currently reported dual-mode sensors for acid phosphatase (ACP) activity are still limited, and most of them more or less exist some deficiencies, such as complicated construction procedure, high-cost, poor biocompatibility, aggregation-caused quenching and limited emission capacity. RESULTS: Herein, we employed Fe3+ functionalized CuInS2/ZnS quantum dots (CIS/ZnS QDs) as nanosensor to develop a novel fluorometric and colorimetric dual-mode assay for ACP activity, combing with ACP-triggered hydrolysis of ascorbic acid 2-phosphate (AAP) into ascorbic acid (AA). The Fe3+ binding to CIS/ZnS QDs can be reduced into Fe2+ during the determination, resulting in the dramatically weakened photoinduced electron transfer (PET) effect and the disappearance of competition absorption. Thus, a highly sensitive ACP assay in the range of 0.22-12.5 U L-1 through fluorescence "turn-on" mode has been achieved with a detection of limit (LOD) of 0.064 U L-1. Meanwhile, the ACP activity can also be quantified by spectrophotometry based on the chromogenic reaction of the formed Fe2+ with 1,10-phenanthroline (Phen). Moreover, the designed nanosensor with good biocompatibility was successfully applied to image and monitor the ACP levels in living cells. SIGNIFICANCE: We believe that the proposed method has remarkable advantages and potential application for ACP assay in terms of the high accuracy, simplicity, low cost, as well as its adequate sensitivity.

The changes in tissue histomorphology and quality of DNA in healthy human autopsied tissues stored at -20 °C and -150 °C.

The aim of this study was to compare the changes in tissue histomorphology and DNA quality in six different healthy tissues (brain, heart, lung, liver, spleen and kidney) exempted during autopsy of healthy individuals and storage at -20 °C and -150 °C three month. Tissue samples were obtained, divided by tissue and temperature group, and for each sample, tissue histomorphology and DNA (isolated from all tissues in duplicated - 72 samples of DNA) quality were analysed. Morphology of tissue samples was studied using H&E staining. DNA was isolated using the phenol-chloroform-isoamyl alcohol method. To assess the concentration and purity of the DNA samples, we used a spectrophotometer to measure absorbance at wavelengths of 280 nm and 260 nm. The fragments of human telomerase reverse transcriptase (hTERT) gene were amplified from the DNA using PCR reaction and then visualised using the 2 % agarose gel. Samples stored at -150 °C sustained the highest degree of histomorphological damage, while samples stored at -20 °C were less degraded, compared to control. The liver samples stored at -20 °C had a mean DNA concentration (1030.4 ± 51.5 ng/µl) higher than the samples of liver tissue stored at -150 °C (497.4 ± 167.1 ng/µl) (p < 0.001). Other tissues did not have statistically significantly different DNA concentration at both temperatures. Liver samples at -20 °C had degraded DNA, showed as the absence of hTERT gene in most of samples. Other tissue samples in both temperature groups had unfragmented DNA. Storing tissue samples at -20 °C is not inferior in terms of DNA yield and integrity, and possibly superior for tissue histomorphology, comparing with samples stored at -150°C.