Development of a magnetic nanocomposite sorbent (NiCoMn/Fe3O4@C) for efficient extraction of methylene blue and Auramine O.

A rapid and efficient method for the simultaneous monitoring and recovery of Auramine O (AO) and Methylene Blue (MB) dyes from water samples is presented. This method, named ultrasound-assisted dispersive-magnetic nanocomposites-solid-phase microextraction (UA-DMN-µSPE), utilizes NiCoMn/Fe3O4@C composite sorbents. Response surface methodology (RSM) combined with artificial neural networks (ANN) and generalized regression artificial neural network (GRNN) under central composite design (CCD) was employed to optimize various parameters for efficient extraction, followed by further refinement using desirability function analysis (DFA) and genetic algorithms (GA). Under optimized conditions, the method achieved exceptional recovery rates (99.5 ± 1.2% for AO and 99.8 ± 1.1% for MB) with acetone as the eluent. Additionally, a high preconcentration factor of 45.50 and 47.30 for AO and MB, respectively, was obtained. Low detection limits of 0.45 ng mL⁻1 (AO) and 1.80 ng mL⁻1 (MB) were achieved with wide linear response ranges (5-1000 and 5-2000 ng mL⁻1 for AO and MB, respectively). The method exhibited good stability with RSDs below 3% for five recycling runs, and minimal interference from various ions was observed. This UA-DMN-µSPE-UV/Vis method offers significant advantages in terms of efficiency, preconcentration, and detection limits, making it a valuable tool for the analysis of AO and MB in water samples.

Eco-friendly phytofabrication of Ficus Benjamina L. based ZnO-doped g-C3N4 nanocomposites for remarkable photocatalysis and antibacterial applications.

The research reported here emphasizes the phytoextract route synthesized ZnO-doped g-C3N4 (GCN) for its photocatalytic activity, which helps to ensure a sustained & healthy environment. The leaf extract solution of Ficus Benjamina L. was used for the synthesis of ZnO nanoparticles, and GCN was prepared via urea using a thermal polymerization process. The flower extract functions as both stabilizers and capping agents during the process of synthesis of ZnO nanoparticles. The synthesized nanocomposites were then calcined at 400 °C and were further characterized with spectroscopy (UV-Vis), diffracted pattern (XRD), and infrared spectroscopy (FTIR). Further, the photocatalytic activity of auramine orange (AO) and methylene blue (MB) dye from phytoextract route synthesized pure ZnO NPs, GCN-Pure, and composites with varied millimolar concentrations of ZnO nanoparticles with GCN of the constant amount was checked. After the complete analysis, it was observed that the series that was prepared of ZnO-GCN nanocomposites showed notable enhancement in the degradation pattern of the methylene blue dye. Apparently, 1.5 mmol (mM) ZnO-GCN presented greater degradation patterns for Auramine orange and Methylene blue dye as compared to other nanocomposites that were synthesized. The observed increased photocatalytic activity has a conceivable explanation. The antibacterial activity studies of the prepared nanocomposites were also performed against the E. coli strain showing an enhanced zone of inhibition towards it.

Paper-based visualization of auramine O in food and drug samples with carbon dots-incorporated fluorescent microspheres as sensing element.

A paper-based assay for visualization of auramine O (AO) was for the first time established by using CFMs as a ratiometric fluorescent probe (RFP). The CFMs were melamine formaldehyde microspheres (MFMs) incorporated with carbon dots (CDs), where the CDs species as sensing units and MFMs as a signal amplification carrier. The proposed RFP can quantitatively measure AO content from 0.0 to 10.0 µM and exhibited an ultralow limit of detection (LOD, 15.7 nM). In particular, obvious luminescence color change of CFMs from blue to green was perceived with naked-eyes and therefore, a solution-based and a paper-based visualization platform were respectively proposed for on-site visual detection of AO with LODs of 1.15 µM and 0.83 µM, separately. Finally, those fluorescence methods were adopted in sensitively quantitative measurement of AO within various food and drug samples, providing new prospects for analysts and technical support in food quality monitoring.

Use of light-emitting diode fluorescence microscopy to detect acid-fast bacilli in sputum as proficient alternative tool in the diagnosis of pulmonary tuberculosis in countries with limited resource settings.

Background: Despite recent advances in the development of more sensitive technologies for the diagnosis of tuberculosis (TB), in resource-limited settings, the diagnosis continues to rely on sputum smear microscopy. This is because smear microscopy is simple, cost-efficient and the most accessible tool for the diagnosis of TB. Our study evaluated the performance of light-emitting diode fluorescence microscopy (LED-FM) using auramine/rhodamine (auramine) and the fluorescein di-acetate (FDA) vital stain in the diagnostic of pulmonary TB in Bamako, Mali. Methods: Sputum smear microscopy was conducted using the FDA and auramine/rhodamine staining procedures on fresh samples using LED-FM to evaluate the Mycobacterium TB (MTB) metabolic activity and to predict contagiousness. Mycobacterial culture assay was utilized as a gold standard method. Results: Out of 1401 TB suspected patients, 1354 (96.65%) were retrieved from database, which were MTB complex culture positive, and 47 (3.40%) were culture negative (no mycobacterial growth observed). Out of the 1354 included patients, 1343 (95.86%), were acid-fast bacillus (AFB) positive after direct FDA staining, 1352 (96.50%) AFB positive after direct Auramine, and 1354 (96.65%) AFB positive with indirect auramine after digestion and centrifugation. Overall, the FDA staining method has a sensitivity of 98.82%, while the sensitivity of Auramine with direct observation was 99.48%, and 99.56% with the indirect examination. Conclusion: This study showed that, using fresh sputum both auramine/rhodamine and FDA are highly sensitive methods in diagnosing pulmonary TB and could be easily used in countries with limited resource settings.

Biodegradable tamarind kernel powder/kappa-carrageenan hydrogel for efficient removal of cationic dyes from effluents.

As an adsorbent for wastewater treatment, an eco-friendly biodegradable hydrogel was utilized in an effort to reduce the hydrogel production costs and decrease its negative impact on the environment. The biodegradable hydrogel of natural polysaccharides, tamarind kernel powder (tkp) with kappa-carrageenan (kcg), was employed as an adsorbent to remove cationic dyes from an aqueous environment. The impact of factors such as initial adsorbate concentration, pH, contact time, temperature, and adsorbent dosage on maximum adsorption were investigated. The tkp-kcg hydrogel has a remarkable swelling percentage of 1840%. The high water penetration of the tkp-kcg hydrogel made the internal adsorption sites for safranin (SF) and auramine-O (AO) dye adsorption accessible. The correlation coefficient supported the Langmuir isotherm model's applicability, with maximum adsorption efficiency of 9.372 mg g-1 for SF and 9.225 mg g-1 for AO. The kinetics of adsorption revealed a pseudo-second order process. Adsorption was an exothermic and spontaneous, according to thermodynamic analyses. Furthermore, the adsorbent was effectively used for five consecutive cycles of SF and AO dye adsorption-desorption. The biodegradation of tkp-kcg hydrogel was characterized by percentage of weight loss, Fourier transform infrared and scanning electron microscopy techniques. The composting technique of biodegradation was used in the biodegradation studies. Using the composting process, 92.6% of the synthesized hydrogel was degraded after 70 days. The results demonstrated that the hydrogel has a high microbiological biodegradability. It is believed that the tkp-kcg hydrogel could have excellent wastewater and agricultural applications due to its high water absorption, excellent retention capacities, cost-effective and ecofriendly synthesis. PRACTITIONER POINTS: Microwave assisted tkp-kcg hydrogel was synthesized with the swelling percentage of 1840%. Synthesized hydrogel showed excellent cationic dyes (SF and AO) adsorption capacity with a good recyclability. The synthesized hydrogel showed very good biodegradability of 92.6 % in 70 days by using composite method.

Comparison of conventional diagnostic methods with molecular method for the diagnosis of pulmonary tuberculosis.

BACKGROUND: Tuberculosis remains one of the deadliest communicable diseases. Prompt diagnosis of active tuberculosis cases facilitates timely therapeutic intervention and minimizes the community transmission. Although conventional microscopy has low sensitivity, still it remains the corner stone for the diagnosis of pulmonary tuberculosis in high burden countries like India. On the other hand, Nucleic acid amplification techniques due to their rapidity and sensitivity, not only help in early diagnosis and management of tuberculosis but also curtail the transmission of the disease. This study therefore was aimed at assessing the diagnostic performance of Microscopy by Ziehl Neelsen (ZN) and Auramine Staining (AO) with Gene Xpert/CBNAAT (Cartridge based nucleic acid amplification test) in the diagnosis of Pulmonary Tuberculosis. METHODS: A prospective comparative study was done on the sputum samples of 1583 adult patients from November 2018 to May 2020 suspected of having pulmonary tuberculosis as per NTEP criteria visiting the Designated Microscopic Centre of SGT Medical College, Budhera, Gurugram. Each sample was subjected to ZN staining, AO staining, and was run on CBNAAT as per National Tuberculosis Elimination Program (NTEP) guidelines. The sensitivity, specificity, PPV and NPV and Area under the curve of ZN microscopy and Fluorescent Microscopy were calculated taking CBNAAT as reference in absence of culture. RESULTS: Out of the 1583 samples studied, 145 (9.15%) and 197 (12.44%) were positive by ZN and AO staining methods respectively. By CBNAAT 246 (15.54%) samples were positive for M. tuberculosis. AO was also able to detect more pauci-bacillary cases than ZN. While CBNAAT detected M. tuberculosis in 49 sputum samples which were missed by both methods of microscopy. On the other hand there were 9 samples which were positive for AFB by both the smear microscopy techniques but M. tuberculosis was not detected by CBNAAT, these were considered as Non-Tuberculous Mycobacteria. Seventeen samples were resistant to rifampicin. CONCLUSION: Auramine Staining technique is more sensitive and less time consuming for the diagnosis of pulmonary tuberculosis as compared to the conventional ZN Staining. CBNAAT can be a useful tool for early diagnosis of patients with high clinical suspicion of pulmonary tuberculosis and detecting rifampicin resistance.

Amazon raw clay as a precursor of a clay-based adsorbent: experimental study and DFT analysis for the adsorption of Basic Yellow 2 dye.

A clay-based adsorbent (CBA) was purified from a sustainable precursor (raw clay, RC), which was obtained from the Amazon region in Brazil. The CBA was characterized using X-ray diffraction (XRD), Fourier Transform Infrared spectroscopy (FTIR), Brunauer-Emmet-Teller surface area (SBET, RC = 23.386 m2.g-1, CBA = 33.020 m2.g-1), Scanning Electron Microscopy (SEM), Energy Dispersive X-ray Spectroscopy (EDS), thermogravimetric analysis (TGA), cation exchange capacity (CEC, CBA = 44.75 cmol/kg), and point of zero charge analyses (pHPZC, CBA = 2.20). Subsequently, CBA was used to adsorb basic yellow 2 (BY2) dye from aqueous solutions. A CBA dosage (1 g/L), initial concentration of dye (C0 = 15 mg/L), and pH (5.6) were ideal conditions for the BY2 dye removal of ~ 98%. The BY2 kinetics was better represented by the pseudo-first-order (PFO) model while the BY2 equilibrium was well represented by the Sips model, with a maximum adsorption capacity of qms = 18.04 mg/g at 28 °C. The negative values of ΔG° and ΔH° showed that the studied process is spontaneous and exothermic, while the values of isosteric heat (∆Hst, -16 to -20 kJ/mol) suggest a predominance of physical interactions. The molecular chemical reactivity of BY2 was investigated using quantum chemical descriptors calculated based on Density Functional Theory (DFT) optimization of the dye molecule, and the results revealed a large energy gap value (4.3900 eV) and considerable chemical hardness (η = 2.1950 eV). Therefore, the correlation between DFT and experimental results consistently sustains that BY2 dye tends to be adsorbed on the CBA surface by electrostatic interactions, thus, this is the possible adsorption mechanism of this process.

A novel and simple heat-based method eliminates the highly detrimental effect of xylene deparaffinization on acid-fast stains.

OBJECTIVES: Histopathology is an important method for diagnosing extrapulmonary tuberculosis, yet tissue sections are often negative for mycobacteria after use of acid-fast stain (AFS). This study investigated the mechanism of AFS use and the detrimental effect of histologic processing-in particular, xylene deparaffinization-on AFS and mycobacterial detection. METHODS: The target of the fluorescent Auramine O (AuO) AFS was investigated using triple staining with DNA- and RNA-specific dyes. The effect of xylene deparaffinization on the acid fastness of mycobacteria in cultures or tissue sections was studied using AuO fluorescence as a quantitative marker. The xylene method was compared with a novel, solvent-free projected-hot-air deparaffinization (PHAD). RESULTS: Co-localization of AuO with DNA/RNA stains suggests that intracellular nucleic acids are the true target of AFS, producing highly specific patterns. Xylene reduces mycobacterial fluorescence significantly (P < .0001; moderate effect size, r = 0.33). The PHAD process yielded significantly higher fluorescence than xylene deparaffinization in tissues (P < .0001; large effect size, r = 0.85). CONCLUSIONS: Auramine O can be applied for nucleic acid staining of mycobacteria in tissues producing typical beaded patterns. Acid-fast staining depends heavily on the integrity of the mycobacterial cell wall, which xylene appears to damage. A solvent-free tissue deparaffinization method has the potential to increase mycobacterial detection significantly.

Behavior of Auramine O in the Aqueous Solution of Two Kolliphors and Their Mixture.

The studies on the behavior of Auramine O (AuO) at the water-air interface and in the bulk phase of the aqueous solution of Kolliphor® ELP (ELP) and Kolliphor® RH 40 (RH40) and their mixture were based on the results obtained from the measurements of the contact angle of water, formamide and diiodomethane on the polytetrafluoroethylene covered by the AuO layer, the surface tension of the aqueous solution of AuO, AuO + ELP, AuO + RH40, AuO + ELP + RH40, density and fluorescence intensity. Based on the obtained results, it was possible to determine components and parameters of the AuO surface tension, concentration and composition of the mixed monolayer, including AuO, ELP and RH40, as well as that of the mixed micelles, and to determine the Gibbs standard free energy of adsorption, micellization and AuO solubilization. The obtained results also showed that surface tension isotherms of the studied solutions can be described by the Szyszkowski equation and the exponential function of the second order and predicted by the Fainerman and Miller equation. In addition, the mixed surface layer composition can be predicted based on the contribution of the components of this layer to the water surface tension reduction.

Evaluation of MetaSystems Automated Fluorescent Microscopy System for the Machine-Assisted Detection of Acid-Fast Bacilli in Clinical Samples.

Manual reading of fluorescent acid-fast bacilli (AFB) microscopy slides is time-intensive and technically demanding. The aim of this study was to evaluate the accuracy of MetaSystems' automated fluorescent AFB slide scanner and analyzer. Auramine O-stained slides corresponding to 133 culture-positive and 363 culture-negative respiratory (n = 284), tissue (n = 120), body fluid (n = 81), and other (n = 11) sources were evaluated with the MetaSystems Mycobacteria Scanner running the NEON Metafer AFB Module. The sensitivity and specificity of the MetaSystems platform was measured as a standalone diagnostic and as an assistant to technologists to review positive images. Culture results were used as the reference method. The MetaSystems platform failed to scan 57 (11.5%) slides. The MetaSystems platform used as a standalone had a sensitivity of 97.0% (129/133; 95% CI 92.5 to 99.2) and specificity of 12.7% (46/363; 95% CI 9.4 to 16.5). When positive scans were used to assist technologists, the MetaSystems platform had a sensitivity of 70.7% (94/133; 95% CI 62.2 to 78.3) and specificity of 89.0% (323/363; 95% CI 85.3 to 92.0). The manual microscopy method had a sensitivity of 79.7% (106/133; 95% CI 71.9 to 86.2) and specificity of 98.6% (358/363; 95% CI 96.8 to 99.6). The sensitivity of the MetaSystems platform was not impacted by smear grade or mycobacterial species. The majority (70.3%) of false positive smears had ≥2+ smear results with the MetaSystems platform. Further performance improvements are needed before the MetaSystems' automated fluorescent AFB slide reader can be used to assist microscopist in the clinical laboratory.

Dispersive solid phase microextraction based on magnesium oxide nanoparticles for preconcentration of auramine O and methylene blue from water samples.

In this study, we investigated the process of preconcentrate and determine trace amounts of Auramine O (AO) and methylene blue (MB) dyes in environmental water samples. For this purpose, the ultrasound-assisted dispersive-magnetic nanocomposites-solid-phase microextraction (UA-DMNSPME) method was performed to extract AO and MB from aqueous samples by applying magnesium oxide nanoparticles (MgO-NPs). The proposed technique is low-cost, facile, fast, and compatible with many existing instrumental methods. Parameters affecting the extraction of AO and MB were optimized using response surface methodology (RSM). Short extraction time, low experimental tests, low consumption of organic solvent, low limits of detection (LOD), and high preconcentration factor (PF) was the advantages of method. The PF was 44.5, and LOD for AO and MB was 0.33 ng mL-1 and 1.66 ng mL-1, respectively. The linear range of this method for AO and MB were 1-1000 ng mL-1 and 5-2000 ng mL-1, respectively. In addition, the relative standard deviation (RSD; n = 5) of the mentioned analytes was between 2.9% and 3.1%. The adsorption-desorption studies showed that the efficiency of adsorbent extraction had not declined significantly up to 6 recycling runs, and the adsorbent could be used several times. The interference studies revealed that the presence of different ions did not interfere substantially with the extraction and determination of AO and MB. Therefore, UA-DMNSPME-UV/Vis method can be proposed as an efficient method for preconcentration and extraction of AO and MB from water and wastewater samples.

Separation and determination of basic orange II, acid orange II and auramine O in soybean products based on ionic liquid reverse micelle microextraction and ultra-high-performance liquid chromatography.

A new analytical method was developed for the separation and determination of basic orange II, acid orange II and auramine O in soybean products. The technique was focused on ionic liquid reverse micelle microextraction (IL-RMME) followed by analysis and determination by ultra-high performance liquid chromatography (UHPLC) with photodiode array detector of three chemical dyes. In this method, IL-RMME solution consisting of ionic liquid [Omim]BF6 and surfactant GenapolX-080 was used as extractant. Important parameters affecting IL-RMME efficiency, such as extraction solvent type and volume, sample solution pH, salt effect, centrifugation speed and time were investigated. Under the optimal condition, the linearity of the method was in the range of 0.1-10 ng mL-1with correlation coefficient above 0.9994 and the limits of detection below 0.03 ng mL-1. At the same time, relative standard deviations of the developed procedure for intra- (n = 5) and inter-day (n = 5) precision were in the range of 5.04-8.50%. The results demonstrated that a simple fast environmentally friendly efficient method was successfully applied in the separation and determination of three chemical dyes in soybean products.

Experimental modelling studies on the removal of dyes and heavy metal ions using ZnFe2O4 nanoparticles.

The presence of dyes and heavy metals in water sources as pollutants is harmful to human and animal health. Therefore, this study aimed to evaluate the efficacy of zinc ferrite (ZnFe2O4) nanoparticles (ZF-NPs) due to their outstanding properties including cost-effectiveness, availability, and applicability for removal of auramine O (AO), methylene blue (MB), and Cd (II). The effect of the main operating parameters such as AO concentration, MB concentration, Cd (II) concentration, adsorbent amount, solution pH, and sonication time was optimized by the response surface methodology (RSM). Optimal conditions were obtained at adsorbent amount of 0.25 g, pH = 6, sonication time of 15 min, and concentration of 15 mg L-1, and more than 91.56% were removed from all three analytes. The adsorption of AO, MB, and Cd (II) onto ZF-NPs followed pseudo-second-order kinetics and the equilibrium data fitted well with Langmuir isotherm. The maximum adsorption capacities of ZF-NPs for AO, MB and Cd (II) were as high as 201.29 mg g-1, 256.76 mg g-1 and 152.48 mg g-1, respectively. Also, the reuse of the adsorbent was investigated, and it was found that the adsorbent can be used for up to five cycles. Based on the results of interference studies, it was found that different ions do not have a significant effect on the removal of AO, MB, and Cd (II) in optimal conditions. The ZF-NPs was investigated successfully to remove AO, MB, and Cd (II) from environmental water samples. The results of this study showed that ZF-NPs can be used as a suitable adsorbent to remove AO, MB, and Cd (II) from aqueous solution.

Label-free DNAzyme for highly sensitive detection of multiple biomolecules in real samples through target-triggered catalytic cleavage reactions with auramine O's discriminated fluorescence emission.

A universal enzyme strand (E-DNA) recyclable L-histidine (L-His), melamine (MA), and cisplatin (CP) biosensor was fabricated on the basis of a target-specific RNA-cleaving DNAzyme with specific auramine O (AuO) dye instead of thioflavin T. In this strategy, the substrate strand (S-DNA) of the RNA-cleaving site was constructed as an intramolecular stem-loop structure, and a GT-rich sequence was imprisoned in the double-stranded stem which inhibits the formation of stable G-quadruplex (G4cpx) with AuO. The presence of L-His initiates a catalytic reaction for cleaving the RNA site of the S-DNA hydrolytically releasing the GT-rich portion, which subsequently combines with AuO and forms a G4cpx for enhanced fluorescent signal. The subsequent addition of MA uncoils the G4cpx to form T-MA-T dsDNA, or addition of CP unwound the G4cpx to form CP-DNA leading to an intensive decrease of AuO emission. Remarkably, the liberated L-His can ultimately cause several rounds of cleavage, and the liberated E-DNA can catalyze the subsequent reaction with the other S-DNA. The use of L-His and E-DNA repeatedly induces S-DNA cleavage and intensifies the emission signal. The results show that the proposed biosensor is extremely sensitive to L-His, MA, and CP with a detection limit of 0.98, 10, and 3.4 nM respectively. To the best of our knowledge, the utilization of AuO as the G4cpx inducer and stabilizer for L-His, MA, and CP detection in real milk and urine samples has never been reported.

Hybrid of sodium polytungstate polyoxometalate supported by the green substrate for photocatalytic degradation of auramine-O dye.

Nowadays, textile industries have severely polluted the ecosystem and water sources via disposal of highly thermo- and photo-stable dyes within the ecology that require practical strategies to remove them from nature. In studies, the photocatalytic disinfection technique has been shown to have widespread applications in indoor air, environmental health, detection, biological, biomedical, laboratory hospital, pharmaceutical food industry, plant safety, waste water, effluents disposal, and drinking water disinfection. Herein, the sodium polytungstate (SPT) polyoxometalate (POM) was synthesized through a multi-step production procedure and hence modified via employing a green protocol by using tartaric acid, glutamic acid, and kombucha solvent toward efficient and total complete removal of the highly toxic, stable, and carcinogenic auramine-O (AO) dye from aqueous media. In this regard, developed materials were well-characterized, and their photocatalysis performance for photodegradation of AO dye was examined. Achieved results showed that the optimum absorption conditions were achieved at pH of 5.0, 15 mg/L of AO concentration, 0.04 g of photocatalyst dosage, and 110 min irradiation time, where SPT and modified SPT via green protocol showed full desirability (desirability function (DF) index of 1) along with 71.75 and 100% removal percentage, respectively. Obtained results justified the superior photocatalytic role of the SPT POM and its derived nanocluster that can be used for the complete removal of highly stable dyes from aqueous media till reaching the drinking water standard.

DNA-Templated Modulation in the Photophysical Properties of a Fluorescent Molecular Rotor Auramine O by Varying the DNA Composition.

This work delineates an integrative approach combining spectroscopic and computational studies to decipher the association-induced fluorescence properties of a fluorescent molecular rotor, viz., auramine O (AuO), after interacting with 20-mer duplex DNA having diverse well-matched base pairs. While exploring the scarcely explored sequence-dependent interaction mechanism of AuO and DNA, we observed that DNA could act as a conducive scaffold to the formation of AuO dimer through noncovalent interactions at lower molecular density. The photophysical properties of AuO depend on the nucleotide compositions as described from sequence-dependent shifting in the emission and absorption maxima. Furthermore, we explored such DNA base pair-dependent fluorescence spectral characteristics of AuO toward discriminating the thermodynamically most stable single nucleotide mismatch in a 20-mer sequence. Our results are interesting and could be useful in developing analogues with further enhanced emission properties toward mismatched DNA sequences.