Effect of adding ytterbium trifluoride filler particles on the mechanical, physicochemical and biological properties of methacrylate-based experimental resins for 3D printing.

OBJECTIVE: To formulate an experimental methacrylate-based photo-polymerizable resin for 3D printing with ytterbium trifluoride as filler and to evaluate the mechanical, physicochemical, and biological properties. METHODS: Resin matrix was formulated with 60 wt% UDMA, 40 wt% TEGDMA, 1 wt% TPO, and 0.01 wt% BHT. Ytterbium Trifluoride was added in concentrations of 1 (G1 %), 2 (G2 %), 3 (G3 %), 4 (G4 %), and 5 (G5 %) wt%. One group remained without filler addition as control (GC). The samples were designed in 3D builder software and printed using a UV-DLP 3D printer. The samples were ultrasonicated with isopropanol and UV cured for 60 min. The resins were tested for degree of conversion (DC), flexural strength, Knoop microhardness, softening in solvent, radiopacity, colorimetric analysis, and cytotoxicity (MTT and SRB). RESULTS: Post-polymerization increased the degree of conversion of all groups (p < 0.05). G2 % showed the highest DC after post-polymerization. G2 % showed no differences in flexural strength from the G1 % and GC (p > 0.05). All groups showed a hardness reduction after solvent immersion. No statistical difference was found in radiopacity, softening in solvent (ΔKHN%), colorimetric spectrophotometry, and cytotoxicity (MTT) (p > 0.05). G1 % showed reduced cell viability for SRB assay (p < 0.05). SIGNIFICANCE: It was possible to produce an experimental photo-polymerizable 3D printable resin with the addition of 2 % ytterbium trifluoride as filler without compromising the mechanical, physicochemical, and biological properties, comparable to the current provisional materials.

Microplate-based 3D-printed image box for urea determination in milk by digital image colorimetry.

In this study, we describe a rapid and high-throughput smartphone-based digital colorimetric method for determining urea in milk. A compact and cost-effective 3D-printed image box microplate-based system was designed to measure multiple samples simultaneously, using minimal sample and reagent volumes. The apparatus was applied for the quantification of urea in milk based on its reaction with p-dimethylaminobenzaldehyde (DMAB). The predictive performance of calibration was evaluated using RGB and different colour models (CMYK, HSV, and CIELAB), with the average blue (B) values of the RGB selected as the analytical signal for urea quantification. Under optimized conditions, a urea concentration linear range from 50 to 400 mg L-1 was observed, with a limit of detection (LOD) of 15 mg L-1. The values found with the smartphone-based DIC procedure are in good agreement with spectrophotometric (spectrophotometer and microplate treader) and reference method (mid-infrared spectroscopy) values. This proposed approach offers an accessible and efficient solution for digital image colorimetry, with potential applications for various target analytes in milk and other fields requiring high-throughput colorimetric analysis.

Development and evaluation of a colorimetric LAMP based-assay targeting the Bacteroides HF183 marker for tracking sewage pollution in environmental waters.

Surface waters are vulnerable to contamination by human and animal feces, posing risks to human health due to potential exposure to enteric pathogens. This research developed a colorimetric loop-mediated isothermal amplification (cLAMP) assay to detect sewage associated Bacteroides dorei HF183/BacR287 (HF183) marker in wastewater and environmental water samples. The host sensitivity and host specificity of the assay were evaluated, and their performance was compared to the Bacteroides HF183 qPCR assay using control materials (gBlocks), environmental water samples seeded with untreated sewage, and ambient environmental water samples. In serial dilutions of control materials, qPCR produced quantifiable data across all dilutions, while cLAMP detected the marker down to 0.001 pg/µL of control materials, which was two orders of magnitude less sensitive than qPCR. All untreated sewage samples (n = 12) tested positive for HF183 by both the qPCR and cLAMP assays, demonstrating a host sensitivity value of 1.00 (maximum value of 1.00). The host specificity by analysing 70 non-human fecal nucleic acid samples revealed cLAMP's specificity value of 0.81 compared to qPCR's 0.64. When testing sewage-seeded environmental water samples, both methods detected HF183 for the lowest amount of sewage, indicating similar detection sensitivity. The application of cLAMP for tracking sewage pollution in environmental waters showed promising results, with moderate agreement between cLAMP and qPCR (κ = 0.510). However, cLAMP occasionally missed detections compared to qPCR, particularly in low-concentration samples. Overall, the cLAMP HF183 assay demonstrated promising potential as a rapid and sensitive method for detecting sewage pollution, offering a viable alternative to qPCR in certain environmental monitoring scenarios.

Aptamer-associated colorimetric reverse transcription loop-mediated isothermal amplification assay for detection of dengue virus.

Current diagnostic methods for dengue, such as serological tests, have limitations in terms of cross-reactivity with other viruses. To address this issue, we explored the potential of combining the loop-mediated isothermal amplification (LAMP) technique with the affinity of aptamers to develop point-of-care testing. In this study, we utilized 60 serum samples. An aptamer capable of binding to the dengue virus was employed as a platform for capturing genetic material, and its performance was compared to a commercial kit. Dengue virus was detected through RT-PCR and colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP), allowing visual observation of the results without the need for equipment. In the context of the aptamer LAMP assay, our analysis revealed the detection of the dengue virus in 38 out of 60 samples, with 95% sensitivity and 100% specificity compared to RT-PCR and/or APTA-RT-PCR. Importantly, we observed no cross-reaction when assessing samples positive for the zika virus, underscoring the assay's selectivity. This innovative aptameric capture of the viral RNA in combination with the RT-LAMP (APTA-RT-LAMP) method has the potential to offer valuable molecular insights into neglected infectious diseases in a simpler and faster manner. IMPORTANCE: Dengue is a neglected tropical disease of significant epidemiological importance in tropical and subtropical countries. Current diagnostics for this infection present challenges, such as cross-reactivity in serological tests. Finding ways to enhance the diagnosis of this disease is crucial, given the absence of specific treatments. An accurate, simple, and effective diagnosis contributes to the improved management of infected individuals. In this context, our work combines molecular biology techniques, such as isothermal loop amplification, with aptamers to detect the dengue virus in biological samples. Our method produces colorimetric results based on a color change, with outcomes available in less than 2 hours. Moreover, it requires simpler equipment compared to molecular PCR tests.

A multicommuted system using bacterial cellulose for urease immobilization and copper (II)-MOF colorimetric sensor for urea spectrophotometric determination in milk.

This work describes determining urea in milk samples using a multicommuted approach with a urease enzyme immobilized in bacterial cellulose and solid MOF as a colorimetric reagent. The Cu(2+)-MOF was characterized by FTIR spectroscopy, XRD, and SEM. The urea quantification was based on the urea hydrolysis reaction catalyzed by urease and reacted with Cu(2+)-MOF forming [Cu(NH3)4]2+, monitored at 450 nm. Linear responses were obtained from 1.0 to 50.0 mg dL-1 urea (R = 0.9959, n = 11), detection and quantitation limits of 0.082 mg dL-1 and 0.272 mg dL-1 respectively, analytical frequency of 8 determinations per hour, 0.8 mL sample solution consumption. Potential interfering studies have shown the selectivity of the proposed method. Addition and recovery tests were performed obtaining variation from 90 to 103%. Applying the F-test and t-test, the results showed no significant difference at the 95% confidence level Comparing the proposed and the reference method.

Efficient trihalomethane quantification in drinking water for minimally-equipped water treatment plants labs.

Chlorine is a common disinfectant used in water treatment. However, its reaction with organic matter can lead to the formation of harmful byproducts, such as trihalomethanes (THMs), which are potentially carcinogenic. To address this issue, the aim of this work was to enhance a colorimetric method capable of quantifying THMs in drinking water through UV/Vis Spectrophotometry, using cost-effective equipment, and validate this methodology for the first time according to established validation protocols. The method's innovation involved replacing the solvent pentane with the more common hexane, along with adjusting the heating ramp, elucidating the mechanisms involved in the process. This method involves the reaction between THMs, pyridine, and NaOH to produce a colored compound, which is then monitored through molecular absorption spectroscopy in the visible region. The method was thoroughly validated, achieving a limit of detection of 13.41 µg L-1 and a limit of quantification of 40.65 µg L-1. Recovery assays ranged from 86.1 % to 90.7 %, demonstrating high accuracy. The quality of the linear fit for the analytical curve exceeded R2 > 0.98. The method was applied to real samples, revealing concentrations ranging from 13.58 to 55.46 µg L-1, all way below the legal limit in Brazil (Maximum Contaminant Levels (MCL) = 100 µg L-1). This cost-effective and straightforward method is suitable for integration into water treatment plant laboratories.

Formulation and assessment of biological properties of garcinia indica fruit extract mouthrinse as an adjunct to oral hygiene regimen: an in vitro analysis.

The prevalence of gingivitis is substantial within the general population, necessitating rigorous oral hygiene maintenance. OBJECTIVE: This study assessed a Garcinia indica (GI) fruit extract-based mouthrinse, comparing it to a 0.1% turmeric mouthrinse and a 0.2% Chlorhexidine (CHX) mouthrinse. The evaluation encompassed substantivity, staining potential, antimicrobial efficacy and cytocompatibility. METHODOLOGY: The study employed 182 tooth sections. For antimicrobial analysis, 64 extracted human teeth coated with a polymicrobial biofilm were divided into four groups, each receiving an experimental mouthrinse or serving as a control group with distilled water. Microbial reduction was assessed through colony forming units (CFU). Substantivity was evaluated on 54 human tooth sections using a UV spectrophotometer, while staining potential was examined on 64 tooth sections. Cytocompatibility was tested using colorimetric assay to determine non-toxic levels of 0.2% GI fruit extract, 0.1% Turmeric, and 0.2% CHX. RESULTS: Data were analysed with one-way ANOVA (α=0.05). Cell viability was highly significant (p<0.001) in the 0.2% GI group (64.1±0.29) compared to 0.1% Turmeric (40.2±0.34) and 0.2% CHX (10.95±1.40). For antimicrobial activity, both 0.2% GI (20.18±4.81) and 0.2% CHX (28.22±5.41) exhibited no significant difference (P>0.05) at end of 12 hours. However, 0.1% Turmeric showed minimal CFU reduction (P<0.001). Substantivity results at 360 minutes indicated statistically significant higher mean release rate in 0.1%Turmeric (12.47±5.84 ) when compared to 0.2% GI (5.02±3.04) and 0.2% CHX (4.13±2.25) (p<0.001). The overall discoloration changes (∆E) were more prominent in the 0.2% CHX group (18.65±8.3) compared to 0.2% GI (7.61±2.4) and 0.1% Turmeric (7.32±4.9) (P<0.001). CONCLUSION: This study supports 0.2% GI and 0.1% Turmeric mouth rinses as potential natural alternatives to chemical mouth rinses. These findings highlight viability of these natural supplements in oral healthcare.

Reliability of the color measurement of resin composites using images obtained using a stereoscopic loupe.

This study assessed the reliability of a color measurement method using images obtained from a charge-coupled device (CCD) camera and a stereoscopic loupe. Disc-shaped specimens were created using the composite Filtek Z350 XT (shades DA1, DA2, DA3, and DA4) (n = 3). CIELAB color coordinates of the specimens were measured using the spectrophotometer SP60 over white and black backgrounds. Images of the same specimens were taken using a CCD camera attached to a stereoscopic loupe. The color of the image was measured (red-green-blue [RGB]) using an image processing software and converted to CIELAB coordinates. For each color coordinate, data from images were adjusted using linear regressions predicting those values from SP60. The whiteness index for dentistry (WID) and translucency parameter (TP00) of the specimens as well as the color differences (ΔE00) among pairwise shades were calculated. Data were analyzed via repeated-measures analysis of variance and Tukey's post hoc test (α = 0.05). Images obtained using the loupe tended to be darker and redder than the actual color. Data adjustment resulted in similar WID, ΔE00, and TP00 values to those observed for the spectrophotometer. Differences were observed only for the WID of shade DA3 and ΔE00 for comparing DA1 and DA3 over the black background. However, these differences were not clinically relevant. The use of adjusted data from images taken using a stereoscopic loupe is considered a feasible method for color measurement.

Development and evaluation of a lyophilization protocol for colorimetric RT-LAMP diagnostic assay for COVID-19.

Molecular diagnostics involving nucleic acids (DNA and RNA) are regarded as extremely functional tools. During the 2020 global health crisis, efforts intensified to optimize the production and delivery of molecular diagnostic kits for detecting SARS-CoV-2. During this period, RT-LAMP emerged as a significant focus. However, the thermolability of the reagents used in this technique necessitates special low-temperature infrastructure for transport, storage, and conservation. These requirements limit distribution capacity and necessitate cost-increasing adaptations. Consequently, this report details the development of a lyophilization protocol for reagents in a colorimetric RT-LAMP diagnostic kit to detect SARS-CoV-2, facilitating room-temperature transport and storage. We conducted tests to identify the ideal excipients that maintain the molecular integrity of the reagents and ensure their stability during room-temperature storage and transport. The optimal condition identified involved adding 5% PEG 8000 and 75 mM trehalose to the RT-LAMP reaction, which enabled stability at room temperature for up to 28 days and yielded an analytical and diagnostic sensitivity and specificity of 83.33% and 90%, respectively, for detecting SARS-CoV-2. This study presents the results of a lyophilized colorimetric RT-LAMP COVID-19 detection assay with diagnostic sensitivity and specificity comparable to RT-qPCR, particularly in samples with high viral load.

Highly Sensitive Whole-Cell Mercury Biosensors for Environmental Monitoring.

Whole-cell biosensors could serve as eco-friendly and cost-effective alternatives for detecting potentially toxic bioavailable heavy metals in aquatic environments. However, they often fail to meet practical requirements due to an insufficient limit of detection (LOD) and high background noise. In this study, we designed a synthetic genetic circuit specifically tailored for detecting ionic mercury, which we applied to environmental samples collected from artisanal gold mining sites in Peru. We developed two distinct versions of the biosensor, each utilizing a different reporter protein: a fluorescent biosensor (Mer-RFP) and a colorimetric biosensor (Mer-Blue). Mer-RFP enabled real-time monitoring of the culture's response to mercury samples using a plate reader, whereas Mer-Blue was analysed for colour accumulation at the endpoint using a specially designed, low-cost camera setup for harvested cell pellets. Both biosensors exhibited negligible baseline expression of their respective reporter proteins and responded specifically to HgBr2 in pure water. Mer-RFP demonstrated a linear detection range from 1 nM to 1 µM, whereas Mer-Blue showed a linear range from 2 nM to 125 nM. Our biosensors successfully detected a high concentration of ionic mercury in the reaction bucket where artisanal miners produce a mercury-gold amalgam. However, they did not detect ionic mercury in the water from active mining ponds, indicating a concentration lower than 3.2 nM Hg2+-a result consistent with chemical analysis quantitation. Furthermore, we discuss the potential of Mer-Blue as a practical and affordable monitoring tool, highlighting its stability, reliance on simple visual colorimetry, and the possibility of sensitivity expansion to organic mercury.

Rapid colorimetric polymyxin B microelution directly from positive blood bottles: because patients with serious infections should not have to wait for results of culture-based methodologies.

PURPOSE: To evaluate the performance of the rapid colorimetric polymyxin B microelution (RCPEm) in determining polymyxin B resistance directly from Enterobacterales-positive blood cultures. METHODS: A set volume of positive blood culture bottles (diluted 1:10) was inoculated into a glucose-broth-phenol red solution (NP solution), where a polymyxin B disk was previously eluted (final concentration of 3 µg/mL). Test was read each 1 h for up to 4 h. Color change from red/orange to yellow indicated resistant isolates. Results were compared to the reference method, broth microdilution (BMD), performed from colonies grown on solid media from the same blood culture bottle. RESULTS: One hundred fifty-two Enterobacterales-positive blood cultures were evaluated, 22.4% (34/152) of them resistant to polymyxin B (including 6.6% with borderline MICs). When performing directly from positive blood cultures (RCPEm-BC), specificity and sensitivity were 99.1% and 94.1%, respectively. Of note, 79.4% (27/34) of truly resistant isolates required 3 h of incubation, compared to the 18 ± 2 h incubation that microtiter plates of BMD demand before reading can be performed. CONCLUSIONS: RCPEm directly from blood cultures has great potential to be part of the routine of clinical microbiology laboratories to establish polymyxin B susceptibility, impacting outcome of patients with bloodstream infections caused by carbapenem-resistant Enterobacterales.

Digital Photocolorimetric Analysis of In Vitro Tooth Color Changes.

OBJECTIVE: Tooth color matching is challenging, and digital photocolorimetry using eLABor_aid (eLAB) provides objective evaluation through polarized photographs. However, its comparability with spectrophotometry remains unclear. METHODS AND MATERIALS: Bovine incisor root canals (n=30) were prepared to simulate an incomplete root apex. The teeth were randomly assigned to three groups based on intracanal medication: control (without medication); calcium hydroxide/propylene glycol; and triple-antibiotic paste (n=10 each). Tooth color was assessed using both eLAB and spectrophotometry. Measurements were taken at the crown medio-cervical region on five-time intervals (baseline, 1, 3, 7, and 14 days). Statistical analysis included two-way repeated-measures ANOVA, Sidak post hoc and Pearson's correlation test (α=0.05). RESULTS: No significant differences were observed between the two methods for either medication or follow-ups (p>0.05). Triple-antibiotic paste exhibited higher color variation (p<0.05). After 7 days, all groups presented significant color changes (p<0.05). Moderate to high correlations (R2 from 0.51 to 0.84, p<0.0001) were found between both methods for all groups at all intervals. CONCLUSION: The eLAB is a reliable method for detecting tooth color changes, and its results are comparable to spectrophotometry analysis.

Modificação de uma resina composta Bulk Fill Flow com extrato acetônico da casca da romã / Modification of a Bulk Fill Flow composite resin with acetonic extract from pomegranate peel

Introdução: A cárie dentária ainda se constitui um problema de saúde pública. Áreas adjacentes a restaurações são frequentemente acometidas por cárie. Por mais que as resinas compostas estejam sendo estudadas e melhoradas, ainda não apresentam atividade antimicrobiana. As cascas da romã (Punica Granatum) são um recurso potencial para compostos bioativos como fenólicos, proantocianidinas e flavonoides, além de apresentarem atividade antioxidante e efeito inibitório contra bactérias Gram-negativas e Gram-positivas. Objetivo: modificar a resina composta Opus Bulk Fill Flow (FGM®) com o extrato acetônico da casca da romã em diferentes concentrações e avaliar a rugosidade da superfície e mudança de cor. Metodologia: foi realizada a extração de 5g de casca da romã utilizando 100mL de solvente acetona 70%. Após rotaevaporação, filtragem e liofilização do extrato, este foi macerado, peneirado e pesado em concentrações diferentes a partir da concentração inibitória mínima capaz de inibir o crescimento de Streptococcus mutans ATCC 700610. A resina composta Opus Bulk Fill Flow foi modificada com esse extrato em diferentes concentrações de forma a gerar 5 grupos: Controle 0 µg (n=10), G930 µg (n=10), G1860 µg (n=10), G3730 µg (n=10) e G7460 µg (n=10). Rugosidade (Ra), diferença de cor (ΔE00) e índice de brancura (WID) foram submetidos aos testes de normalidade. Os dados não-paramétricos de Ra foram submetidos ao teste de Kruskal-Wallis com pós-teste de Dunn e os dados paramétricos do ΔE00 e WID foram submetidos ao teste ANOVA 1 Fator com pósteste de Tukey por meio do software GraphPad Prism 8 e Microsoft Excel 2019. Resultados: Verificou-se que a média do índice de brancura (WID) diminuiu conforme o aumento da concentração do extrato na resina modificada (p<0,05), assim como, as resinas modificadas se tornaram, visivelmente a olho nu, um pouco mais amareladas. Após 1 mês, as amostras dos grupos experimentais sofreram a mesma variação de cor (ΔE00) que o grupo controle, uma vez que os valores das médias foram semelhantes. Portanto, todos os grupos apresentaram estabilidade de cor. A rugosidade superficial (Ra) não mostrou diferença estatisticamente significativa (p>0,05). Todos os grupos apresentaram médias com valores similares a 0,06µm. Conclusão: A alteração de cor na resina, com a inserção do extrato, ainda na maior concentração, se manteve na classificação do matiz A. O que não afeta as propriedades organolépticas e pode ser considerada uma cor similar à cor dos dentes naturais. A adição do extrato na resina manteve a rugosidade superficial de todos os grupos dentro do valor ideal, prevenindo a adesão de biofilmes e microrganismos e proporcionando conforto ao toque da língua (AU).

Introduction: Dental caries still constitutes a public health problem. Areas adjacent to restorations are often affected by caries. Even though resin composites are being studied and improved, they still do not have antimicrobial activity. Pomegranate peels (Punica Granatum) are a potential resource for bioactive compounds such as phenolics, proanthocyanidins and flavonoids, in addition to presenting antioxidant activity and inhibitory effects against Gram-negative and Gram-positive bacteria. Objective: to modify the Opus Bulk Fill Flow (FGM™) resin composite with the acetone extract of pomegranate peel in different concentrations and evaluate the surface roughness, and color change. Methodology: 5g of pomegranate peel was extracted using 100mL of 70% acetone solvent. After rotary evaporation, filtering and lyophilization of the extract, it was macerated, sieved and weighed at different concentrations based on the minimum inhibitory concentration capable of inhibiting the growth of Streptococcus mutans ATCC 700610. The Opus Bulk Fill Flow resin composite was modified with this extract in different concentrations to generate 5 groups: Control 0 µg (n=10), G930 µg (n=10), G1860 µg (n=10), G3730 µg (n=10) e G7460 µg (n=10). Roughness (Ra) and color difference (ΔE00) were subjected to normality tests and non-parametric data were subjected to the Kruskal-Wallis test with Dunn's post-test using GraphPad Prism 8 and Microsoft Excel 2019 software. Results: It was found that the average whiteness index (WID) decreased as the concentration of the extract in the modified resin increased (p<0.05), as well as the modified resins became, visibly to the naked eye, a little more yellowish. After 1 month, samples from the experimental groups suffered the same color variation (ΔE00) as the control group, since the average values were similar. Therefore, all groups showed color stability. Surface roughness (Ra) did not show a statistically significant difference (p>0.05). All groups presented average values similar to 0.06µm. Conclusion: The color change in the resin, with the insertion of the extract, even at the highest concentration, remained in the classification of hue A. This does not affect the organoleptic properties and can be considered a color similar to the color of natural teeth. The addition of the extract to the resin composite maintained the surface roughness of all groups within the ideal value, preventing the adhesion of biofilms and microorganisms and providing comfort to the touch of the tongue (AU).

Development and evaluation of a single-tube nested PCR with colorimetric assay for Mycobacterium tuberculosis detection.

Molecular techniques have revolutionized tuberculosis (TB) diagnosis by offering a faster and more sensitive approach, detecting Mycobacterium tuberculosis (Mtb) DNA directly from samples. Single-tube nested PCR (STNPCR) combines two PCR reactions with separate oligonucleotide sets in a single tube. Moreover, colorimetric methods in PCR products have been studied for pathogen detection. Thus, this study aimed to establish a novel system based on colorimetric STNPCR for Mtb detection using microtiter plates with IS6110-amplified fragments. The results showed a general colorimetric STNPCR detection limit of 1 pg/µl. Its general sensitivity and specificity were 76.62 and 60.53%, respectively, with kappa index agreement of 0.166.

A total of 318 biological samples (urine, plasma, peripheral blood mononuclear cells, pleural fluid and sputum) from pulmonary/extrapulmonary TB and non-TB patients were used in this study. The colorimetric STNPCR assay using IS6110 as the target gene was developed and optimized for Mtb detection based on similar validated systems. Cut-off values based on receiver operator characteristic curve analysis were defined to determine the sensitivity and specificity for each sample type. The technique's performance was assessed according to kappa index calculations and interpretation.

Resazurin to determine the minimum inhibitory concentration on antifungal susceptibility assays for Fonsecaea sp. using a modified EUCAST protocol.

Chromoblastomycosis is a fungal chronic disease, which affects humans, especially in cutaneous and subcutaneous tissues. There is no standard treatment for Chromoblastomycosis, and it is a therapeutic challenge, due natural resistance of their causative agents, inadequate response of patients and common cases of relapse. Protocols for determination of antifungal drugs susceptibility are not standardized for chromoblastomycosis agents and endpoint definition is usually based on visual inspection, which depends on the analyst, making it sometimes inaccurate. We presented a colorimetric and quantitative methodology based on resazurin reduction to resofurin to determine the metabolic status of viable cells of Fonsecaea sp. Performing antifungal susceptibility assay by a modified EUCAST protocol allied to resazurin, we validated the method to identify the minimum inhibitory concentrations of itraconazole, fluconazole, amphotericin B, and terbinafine for eight Fonsecaea clinical isolates. According to our data, resazurin is a good indicator of metabolic status of viable cells, including those exposed to antifungal drugs. This work aimed to test resazurin as an indicator of the metabolic activity of Fonsecaea species in susceptibility assays to antifungal drugs. Species of this genus are the main causative agents of Chromoblastomycosis, which affects humans.

Fabrication of paper-based analytical devices using stencil-printed glass varnish barriers for colorimetric detection of salivary α-amylase.

BACKGROUND: Developing disposable paper-based devices has positively impacted analytical science, particularly in developing countries. Some benefits of those devices include their versatility, affordability, environmentally friendly, and the possibility of being integrated with portable electrochemical or colorimetric detectors. Paper-based analytical devices (PADs) comprising circular zones and microfluidic networks have been successfully employed in the analytical chemistry reign. However, the combination of the stencil-printing method and alternative binder has not been satisfactorily explored for fabricating colorimetric paper devices. RESULTS: We developed PADs exploring the stencil printing approach and glass varnish as the hydrophobic chemical agent. As a proof-of-concept, the colorimetric assay of salivary α-amylase (sAA) was performed in saliva samples. Through the scanning electron microscopy measurements, it was possible to indicate satisfactory definitions between native fibers and barrier, and that the measured values for the channel width revealed suitable fidelity (R2 = 0.99) with the nominal widths (ranging from 400 to 5000 µm). The proposed hydrophobic barrier exhibited excellent chemical resistance. The analytical applicability for detecting sAA revealed linear behavior in the range from 2 to 12 U mL-1 (R2 = 0.99), limit of detection of 0.75 U mL-1, reproducibility (RSD ≤2.4%), recovery experiments ranged from 89 to 108% and AGREE response (0.86). In addition, the colorimetric analysis of sAA in four different saliva samples demonstrated levels ranging from 202 to 2080 U mL-1, which enabled monitoring the absence and presence of periodontitis. SIGNIFICANCE: This report has presented the first use of a self-adhesive mask and glass varnish for creating circular zones and microfluidic architectures on paper without using thermic or UV curing treatments. Also, the proposed analytical methodology for detecting sAA exhibited suitable ecological impact considering the AGREE tool. We believe the proposed fabrication of paper devices emerges as a novel, simple, high-fidelity microfluidic channel and portable analytical approach for colorimetric sensing.

Colorimetric LAMP Assay for Detection of Xanthomonas phaseoli pv. manihotis in Cassava Through Genomics: A New Approach to an Old Problem.

Bacterial blight caused by Xanthomonas phaseoli pv. manihotis (Xpm) is considered the main bacterial disease that affects cassava, causing significant losses when not properly managed. In the present study, a fast, sensitive, and easy-to-apply method to detect Xpm via colorimetric loop-mediated isothermal amplification (LAMP) was developed. To ensure the use of a unique-to-the-target pathovar core region for primer design, 74 complete genomic sequences of Xpm together with different bacterial species and pathovars were used for comparative genomics. A total of 42 unique genes were used to design 27 LAMP primer sets, from which nine primers were synthesized, and only one (Xpm_Lp1 primer set) showed sufficient efficiency in preliminary tests. The sensitivity, assessed by a serial dilution of the type strain (IBSBF 278) DNA, yielded high sensitivity, detecting up to 100 fg. The LAMP primers showed high specificity, did not cross-react with other bacterial species or other pathovars tested, and amplified only the Xpm isolates. Tests confirmed the high efficiency of the protocol using infected or inoculated macerated cassava leaves without the need for additional sample treatment. The LAMP test developed in this study was able to detect Xpm in a fast, simple, and sensitive way, and it can be used to monitor the disease under laboratory and field conditions.

Feasibility of using colorimetric devices for whole and ground coffee roasting degrees prediction.

BACKGROUND: Coffee roasting is one of the crucial steps in obtaining a high-quality product as it forms the product's color and flavor characteristics. Roast control is made by visual inspection or traditional instruments such as the Agtron spectrophotometer, which can have high implementation costs. Therefore, the present study evaluated colorimetric approaches (a bench colorimeter, smartphone digital images, and a colorimetric sensor) to predict the Agtron roasting degrees of whole and ground coffee. Two calibration approaches were assessed, that is, multiple linear regression and least-squares support vector machine. For that, 70 samples of whole and ground roasted coffees comprising the Agtron roasting range were prepared. RESULTS: The results showed that all three colorimetric acquisition types were efficient for the model building, but the bench colorimeter and the smartphone digital images generally performed with good determination coefficients and low errors as measured by external validation. For the whole bean coffee, the best model presented a determination coefficient (R2) of 0.99 and a root-mean-squared error (RMSE) of 1.91%, while R2 of 0.99 and RMSE of 0.87% was obtained for ground coffee, both using the colorimeter. CONCLUSION: The obtained models presented good prediction capability, as assessed by external validation and randomization tests. The obtained findings point to an alternative for coffee roasting monitoring that can lead to higher digitalization and local control of the process, even for smaller producers, due to its lower costs. © 2024 Society of Chemical Industry.

Mixed-Mode Solar Drying and its Effect on Physicochemical and Colorimetric Properties of Zompantle (Erythrina Americana).

A mixed-mode solar drying was developed to evaluate the physicochemical and colorimetric properties of Zompantle (Erythrina americana). A 22-factorial design was used; the operation mode (mesh shade and direct) and airflow (natural convection and forced convection) were established as factors in this design. The initial moisture content in the Zompantle flower was reduced from 89.03% (w.b) to values that ranged from 3.84% to 5.84%; depending on the operation mode of the dryer, the final water activity ranged from 0.25 to 0.33. The Zompantle's components as proteins (4.28%), antioxidant activity (18.8%), carbohydrates (4.83%), fat (0.92%), fiber (3.71%), ash (0.94%), and total soluble solids (3°Brix) increased as the water was evaporated during the drying. The increment in the Zompantle's components depends on the operation mode; in direct mode and natural convection, the proteins, antioxidant activity, carbohydrates, fat, fiber, ash, and total soluble solids were 6.99%, 61.69%, 79.05%, 1.20%, 3.84%, 8.70%, and 45 °Brix, respectively. The total drying efficiency was 14.84% with the direct mode and natural convection (DM-NC) and 17.10% with the mesh shade and natural convection (MS-NC). The Hue angle measures the property of the color; the indirect mode and natural convection keep the hue angle close to the initial value (29.2 °). The initial chroma value of the Zompantle flower was 55.07; the indirect mode and natural convection kept high saturation (37.58); these dry conditions ensured a red color in the dehydrated Zompantle. Dehydrated Zompantle's flowers could have several practical applications, such as an additive in traditional Mexican cuisine.

A digital image-based colorimetric method for measuring free acidity in edible vegetable oils.

The standard method used to quantify free acidity (FA) in vegetable oil is neutralization titration, which requires many toxic chemicals and depends on an analyst's experience in detecting endpoints. Here, a digital image colorimetry (DIC) method using a smartphone camera was developed to measure FA in vegetable oils. A cupric acetate solution was used to produce the colorimetric reaction. The coloured solutions were imaged, and R values (from the RGB colour system) were calibrated against the respective FAs in the standards. The FA values of the samples were determined by standard addition calibration. These results were compared to measurements of FA obtained by the standard titrimetric method. An excellent correlation was obtained, with an R2 of 0.98 and a mean absolute error of 0.06%. The chemicals needed for analysis were reduced by approximately 90%. Thus, DIC is a less subjective and more economical method for determining FA in vegetable oils.