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.

Optimal fluorescent-dye staining time for the real-time detection of microbes: a study of Saccharomyces cerevisiae.

AIMS: To provide information on the time-dependent behaviour of microbe staining by fluorescent dyes in the order of seconds, which is important in terms of the recent rapid and online techniques for microbe measurements and/or environmental microbe analysis. METHODS AND RESULTS: For combinations of yeast (Saccharomyces cerevisiae) and typical dyes, including DAPI (4',6-diamidino-2-phenylindole) and Auramine-O, a suspension of yeast cells in ultrapure water was injected into a dye solution in a micro cuvette placed inside a spectrofluorometer and the fluorescence intensity of the resulting solution was measured at 1 s intervals, starting immediately after the mixing and continued until the time for the maximum intensity using various concentrations of yeast and dyes. The relaxation time τ, which corresponds to ~63·2% of the maximum fluorescence intensity, was shown to decrease to below 1 s with increasing DAPI concentration, whereas it remained constant for 2-3 s with increasing Auramine-O concentration, for example at a yeast concentration of 100 µg ml-1 . CONCLUSIONS: For the conditions of yeast >10 µg ml-1 , DAPI >1 µg ml-1 and Auramine-O >0·1 µg ml-1 , τ could be adjusted to below 5 s to achieve a rapid and stable staining. SIGNIFICANCE AND IMPACT OF THE STUDY: Design and operating conditions for rapid and online measurements of microbes can be optimized.

Preparation of a stoichiometric molecularly imprinted polymer for auramine O and application in solid-phase extraction.

We describe a stoichiometric approach to the synthesis of molecularly imprinted polymers specific for auramine O. Using the stoichiometric interaction in molecular imprinting, no excess of binding sites is necessary and binding sites are only located inside the imprinted cavities. The free base of the template was obtained to facilitate the interaction with the monomers. Itaconic acid was selected as the functional monomer, and stoichiometric ratio of the interaction with the free base was investigated. The molecularly imprinted polymer preparation conditions such as cross-linker, molar ratio, porogen were optimized as divinylbenzene, 1:2:20 and chloroform/N,N-dimethylformamide, respectively. Under the optimum conditions, a good imprinting effect and very high selectivity were achieved. A solid-phase extraction method was developed using the molecularly imprinted polymers as a sorbent and extraction procedure was optimized. The solid-phase extraction method showed a high extraction recovery for auramine O in its hydrochloride form and free form compared to its analogues. The results strongly indicated that stoichiometric imprinting is an efficient method for development of high selectivity molecularly imprinted polymers for auramine O.

Preparation of stoichiometric molecularly imprinted polymer coatings on magnetic particles for the selective extraction of auramine O from water.

A novel magnetic molecularly imprinted polymer for the selective recognition of auramine O was rationally designed via screening from a library of nonimprinted polymers. A stoichiometric ratio of functional monomer (itaconic acid) and template molecule (auramine O) was found to be 1.5. Meanwhile, the synthesized SiO2 @Fe3 O4 was modified by 0.5 mol/L hydrochloric acid to facilitate the preparation of magnetic molecularly imprinted polymer particles. Adsorption experiments showed that the magnetic polymer particles exhibited good selectivity, recoveries, and enrichment performance. The stoichiometric imprinted polymers have been employed for the selective preconcentration of auramine O from lake water sample. The high specificity of the stoichiometric imprinted polymers was proven in the extraction of mixture solution of auramine O, auramine O hydrochloride, and chrysoidine, and the recoveries ranged between 99.66 and 108.75% (RSD 2.6-3.7%, n = 3) for lake water. These results suggest that this method is effective and can be successfully applied to the analysis of auramine O in environmental water samples.

Gum xanthan-psyllium-cl-poly(acrylic acid-co-itaconic acid) based adsorbent for effective removal of cationic and anionic dyes: Adsorption isotherms, kinetics and thermodynamic studies.

The present work highlights the synthesis of the adsorbent based on Gum xanthan-psyllium hybrid backbone graft co-polymerized with polyacrylic acid-co-polyitaconic acid chains for the rapid sequestration of auramine-O (Aur-O) and eriochrome black-T (EBT) dyes from the aqueous fluid. The excellent dye removal efficiency of 90.53% for EBT and 95.63% for Aur-O was found at initial dye concentration of 30mgL-1 (EBT) and 15 mgL-1 (Aur-O) 40mL-1 with an adsorbent dose of 600mg within time duration of 5h and 323K temp. The adsorption isotherm data fitted well with Langmuir isotherm and Freundlich isotherm for Aur-O and EBT dyes (R2 ≥ 0.90), respectively. The adsorption kinetics depicted that pseudo-second order kinetics was followed simultaneously with intra-particle diffusion for both the dyes. Thermodynamic parameters such as ΔG°, ΔH° and ΔS° were also calculated and confirmed the spontaneity, randomness and endothermic nature of the adsorption process. Further, the adsorbent exhibited good recyclability efficiency for the capture of Aur-O and EBT from aqueous solution with minimal activity decline after six and three cycles, respectively. So, the synthesized adsorbent could be used successfully by the textile industries for the treatment of dye contaminated water with excellent competency.

Auramine O, an incense smoke ingredient, promotes lung cancer malignancy.

Burning incense to worship deities is a popular religious ritual in large parts of Asia, and is a popular custom affecting more than 1.5 billion adherents. Due to incomplete combustion, burning incense has been well recognized to generate airborne hazards to human health. However, the correlation between burning incense and lung cancer in epidemiological studies remains controversy. Therefore, we speculated that some unknown materials in incense smoke are involved in the initiation or progression of lung cancer. Based on this hypothesis, we identified a major compound auramine O (AuO) from the water-soluble fraction of incense burned condensate using mass spectrometry. AuO is commonly used in incense manufacture as a colorant. Due to thermostable, AuO released from burned incenses becomes an unexpected air pollutant. AuO is classified as a Group 2B chemical by the International Agency of Research on Cancer (IARC), however, the damage of AuO to the respiratory system remains elusive. Our study revealed that AuO has no apparent effect on malignant transformation; but, it dramatically promotes lung cancer malignancy. AuO accumulates in the nucleus and induces the autophagy activity in lung tumor cells. AuO significantly enhances migration and invasive abilities and the in vitro and in vivo stemness features of lung tumor cells through activating the expression of aldehyde dehydrogenase family 1 member A1 (ALDH1A1), and ALDH1A1 knockdown attenuates AuO-induced autophagy activity and blocks AuO-induced lung tumor malignancy. In conclusion, we found that AuO, an ingredient of incense smoke, significantly increases the metastatic abilities and stemness characters of lung tumor cells through the activation of ALDH1A1, which is known to be associated with poor outcome and progression of lung cancer. For public health, reducing or avoiding the use of AuO in incense is recommended.

Examining Diagnostic Efficacy: GeneXpert Versus Traditional Staining Techniques With Culture in the Diagnosis of Tuberculosis.

Introduction The tuberculosis (TB) diagnosis involves various methods, such as microscopic examination, culture-based methods, molecular techniques, chest X-rays, serological tests, and interferon-gamma release assays. These methods help identify and confirm TB and its resistance to rifampicin, balancing speed and accuracy for prompt treatment initiation and effective disease management. Aims and objectives To assess the diagnostic accuracy of GeneXpert, Ziehl-Neelsen staining, and fluorescence staining compared to culture media in TB-suspected patients. Materials and methods We analysed 416 patient samples for TB over one year using GeneXpert, Ziehl-Neelsen staining, fluorescence staining, and Löwenstein-Jensen (LJ) medium. Only samples with a suspicion of TB were included in the study. The samples received without clinical history and requests for all four tests were excluded. Results A total of 416 patient samples were categorised into pulmonary and extrapulmonary samples. GeneXpert detected 62 positive cases for TB, out of which 53 were rifampicin-sensitive, seven were rifampicin-indeterminate, and two were rifampicin-resistant. The indeterminate samples were further evaluated using the line probe assay (LPA), of which six were rifampicin-sensitive, and one was rifampicin-resistant. Fluorescent staining detected 44 cases, Ziehl-Neelsen staining detected 40 cases, and LJ culture medium detected 65 cases. Conclusion GeneXpert is superior to staining methods for detecting TB. GeneXpert, combined with microscopy and culture, can enhance TB and multi-drug resistant tuberculosis (MDR-TB) detection and aid in early treatment initiation.

Preparation and evaluation of a novel molecularly imprinted SPE monolithic capillary column for the determination of auramine O in shrimp.

A novel method combining molecular imprinting and SPE was developed in a capillary column for the determination of auramine O in shrimp. The capillary monolithic column was prepared by UV-initiated in situ polymerization, using auramine O as template and methacrylic acid and ethylene dimethacrylate as functional monomer and cross-linker, respectively. The properties of the prepared capillary monolithic column were investigated under the optimized conditions coupled with HPLC, and then the morphologies of the inner polymers were characterized by SEM. The calibration curve was expressed as A = 103C + 19.8 (r = 0.9992) with a linear range of 0.25-25.0 µg/mL, and the recoveries of auramine O at different concentrations in shrimp ranged from 90.5 to 92.4% with RSDs ranging from 2.1 to 4.4%. The capacities of the molecularly imprinted polymer and nonimprinted polymer columns were 0.722 and 0.147 µg/mg, respectively, and the LOD (S/N = 3) of auramine O in shrimp was 17.85 µg/kg. Under the selected conditions, the enrichment factors obtained were higher than 70-fold. The results indicate that the prepared molecularly imprinted capillary monolithic column was reliable and applicable to the analysis of auramine O in shrimp.

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.

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.

Visible light enhanced photocatalytic degradation of organic pollutants with SiO2/g-C3N4 nanocomposite for environmental applications.

The development of eco-friendly photocatalysts is gaining attention as an effective approach for degrading organic pollutants. In the present study, the composite materials are composed of various components with varying structures that combine to enhance their characteristics and widen their applications. This work uses the hydrothermal method for the fabrication of a novel and steady SiO2/g-C3N4 photocatalyst. The amount of SiO2 was fixed, and graphitic carbon nitride (g-C3N4) was varied in the ratio (1:x, where x = 1, 2, 3) and abbreviated as SCN1, SCN2, and SCN3. The optical properties, surface morphology, and structural analysis of the prepared nanocomposites were studied using various techniques such as FTIR, TGA, X-ray diffraction, and ultraviolet-visible spectroscopy. The results show that SCN2 nanocomposites significantly improved the photocatalytic activity, with a degradation efficiency of 70% for auramine O and 84.6% for xylenol orange dye under visible light irradiation, which is a result of their large surface area and efficient electron-hole separation rate.

Rapid Indentification of Auramine O Dyeing Adulteration in Dendrobium officinale, Saffron and Curcuma by SERS Raman Spectroscopy Combined with SSA-BP Neural Networks Model.

(1) Background: Rapid and accurate determination of the content of the chemical dye Auramine O(AO) in traditional Chinese medicines (TCMs) is critical for controlling the quality of TCMs. (2) Methods: Firstly, various models were developed to detect AO content in Dendrobium officinale (D. officinale). Then, the detection of AO content in Saffron and Curcuma using the D. officinale training set as a calibration model. Finally, Saffron and Curcuma samples were added to the training set of D. officinale to predict the AO content in Saffron and Curcuma using secondary wavelength screening. (3) Results: The results show that the sparrow search algorithm (SSA)-backpropagation (BP) neural network (SSA-BP) model can accurately predict AO content in D. officinale, with Rp2 = 0.962, and RMSEP = 0.080 mg/mL. Some Curcuma samples and Saffron samples were added to the training set and after the secondary feature wavelength screening: The Support Vector Machines (SVM) quantitative model predicted Rp2 fluctuated in the range of 0.780 ± 0.035 for the content of AO in Saffron when 579, 781, 1195, 1363, 1440, 1553 and 1657 cm-1 were selected as characteristic wavelengths; the Partial Least Squares Regression (PLSR) model predicted Rp2 fluctuated in the range of 0.500 ± 0.035 for the content of AO in Curcuma when 579, 811, 1195, 1353, 1440, 1553 and 1635 cm-1 were selected as the characteristic wavelengths. The robustness and generalization performance of the model were improved. (4) Conclusion: In this study, it has been discovered that the combination of surface-enhanced Raman spectroscopy (SERS) and machine learning algorithms can effectively and promptly detect the content of AO in various types of TCMs.

Label free detection of auramine O by G-quadruplex-based fluorescent turn-on strategy.

Auramine o (AO) is a synthetic dye used in paper and textile industries. Although it has been an unauthorized food additive in many countries due to its toxic and carcinogenic possibility, its illegal uses have been detected in certain food products such as pasta, semolina and spices and also in pharmaceuticals. The presence of AO in food products should be monitored, therefore, to minimize the negative health effects on consumers. In this study, a simple, highly sensitive and selective label free detection method was investigated for AO by G-quadruplex-based fluorescent turn-on strategy. The optimum fluorescent detection assay was achieved with a specific G-quadruplex DNA sequence, c-myc, at 400 nM in Tris-HCl buffer at pH 7.4. The linearity of fluorescence intensity depending on AO concentration ranged from 0 to 0.07 µM and LOD and LOQ were 3 nM and 10 nM, respectively. The G-quadruplex-based detection assay was highly specific for AO as compared to other two synthetic food colorings and successfully applied to determine AO in pasta, bulgur and curry powder with recoveries in the range from 70.33% to 106.49%. This G-quadruplex-based label free detection assay has a significant potential to be used in the detection of AO in food products.

Comparative Evaluation of Clinical, Cytological and Microbiological Profile in Abdominal vs. Cervical Lymph Nodal Tuberculosis with Special Emphasis on Utility of Auramine-O Staining.

CONTEXT: Extrapulmonary tuberculosis (EPTB) especially abdominal lymph nodal tuberculosis (LNTB) poses a unique diagnostic challenge. The clinical, cytological, and microbiological profiles, especially with respect to the use and role of Auramine -O (AO) stain, are not as well characterized in abdominal LNTB as cervical LNTB and were evaluated in the present comparative study. SUBJECTS AND METHODS: This study was conducted in the Department of Pathology of a tertiary care hospital in Shillong, Meghalaya in 540 clinical suspected cases of tuberculosis who underwent FNAC. The smears were submitted for Leishman's stain for cytological analysis, along with ZN and Auramine O stain for demonstration of the organism, analyzed, and scored and the results were compared with culture wherever available. The results from abdominal and cervical lymph nodal tuberculosis were compared using Microsoft Excel and SPSS software. RESULTS: Out of 540 cases, most were tuberculosis (266) followed by reactive lymphadenitis (162), malignancy, and acute necrotizing lesion. On comparing, abdominal lymph nodes (n = 163) were more likely to reveal cheesy/purulent material macroscopically, necrotizing lymphadenitis along with ZN stain and Auramine positivity (P < 0.05) while cervical lymph nodes (n = 66) revealed a higher proportion of granulomatous lymphadenitis and culture positivity (P < 0.05). The sensitivity, NPV, and diagnostic accuracy of AO stain (85.9%, 48.0%, and 62.3%) were higher as compared to ZN stain (47.4%, 39.3%, and 51.9%) with culture as the gold standard. The combined sensitivity of Ziehl Neelsen stain and Auramine stain was 92.05%. CONCLUSION: Cytological and microbiologic features of abdominal LNTB differ from cervical LNTB. Moreover, AO stain increases the smear positivity, is almost twice as sensitive as ZN stain and should be used as an adjunct in cytological material wherever available.

Auramine O in foods and spices determined by an UPLC-MS/MS method.

Auramine O (AO) is a banned food additive and has been classified as an illegal colourant. Therefore, the presence of AO in food should be strictly monitored. In this study, a sensitive UPLC-MS/MS method was applied to monitor AO in 211 food and spice samples. The optimised separation was achieved with a mobile phase consisting of 100 mM ammonium formate at pH 2.9 and acetonitrile, reversed-phase CORTECS T3 column (2.7 µm, 2.1 × 100 mm) operated at 40ºC with a gradient time of 20.0 min (0-95% methanol) at a flow rate of 0.3 mL/min. Limit of detection (LOD) and limit of quantification (LOQ) of the method were 0.1 µg/kg and 0.5 µg/kg, respectively. The results showed that 27.0% of samples were contaminated with AO. Considering the common consumption of sour bamboo shout and turmeric powder by so many consumers, AO exposure is significant and should be decreased.

The yield of Auramine O staining using led microscopy with bleach treated sputum samples for detection of pulmonary tuberculosis at St. Peter tuberculosis specialized hospital, Addis Ababa, Ethiopia.

BACKGROUND: Smear microscopy is the mainstay for diagnosis of Tuberculosis (TB) in Ethiopia. This technique; however, is insensitive to detect Mycobacteria from most clinical specimens. Currently, light emitting diode (LED) fluorescence microscope is advocated to be used in high Tuberculosis (TB) burden settings by World Health Organization (WHO). However, the utility of this method is not evaluated for bleach treated sputum samples in Ethiopia. OBJECTIVE: The objective of the study is to evaluate the diagnostic importance of Auramine O (AO) staining in direct and concentrated sputum against conventional Zehil-Neelsen (ZN) and culture from the sputum samples of suspected pulmonary tuberculosis patients. METHODS: A cross-sectional study was conducted on 346 adult new pulmonary TB suspected patients at St. Peter's Specialized Hospital, Addis Ababa, Ethiopia. Three sputum samples (spot-morning-spot) were collected in sterile cups for direct Zehil-Neelsen and AO staining. Morning sputum samples were used for Mycobacterial culture on Mycobacterial Growth Indicator Tube (MGIT) 960. Sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) were evaluated against the gold standard culture method. Data were analyzed using STATA version 13.0. All statistical tests were considered as statistically significant if the two sided P-value was < 0.05. RESULTS: Bleach treated sputum samples with AO staining yielded more cases as compared to direct ZN and direct AO by 6.3% and 11.5%, respectively. The sensitivity of concentrated AO and direct AO were remarkably high as compared to conventional ZN (71.8% vs. 44.5% and 62.7% vs. 44.5%). The concentrated sputum with staining of AO had a high rate (18.6%) of detecting scanty graded smears as compared to conventional ZN method. CONCLUSIONS: Our findings indicated that the concentrated sputum with AO staining yielded high rate of sensitivity (71.8%) as compared to the conventional ZN method (44.5%). Moreover, the concentrated sputum with AO staining had superior ability in detecting scanty graded smears compared to the conventional ZN method. Therefore, it is recommended to utilize AO staining with LED microscopy for better diagnosis of Acid Fast Bacilli (AFB) from TB suspected cases and patients with pauci-bacillary TB in Ethiopia.

Integration of micronucleus, comet, and Pig-a gene mutation endpoints into rat 15-day repeat-treatment studies: Proof-of-principle with Auramine O.

A series of genotoxicity assessments were conducted on male Sprague Dawley rats treated with Auramine O (AO) to establish a multiple-endpoint assay. The rat liver micronucleus assay, in combination with the comet assay, peripheral blood micronucleus assay, and erythrocyte Pig-a assay in the same experiment, comprehensively assess the genotoxicity of AO. Rats were orally exposed to 0, 100, 200, or 400 mg/kg/day AO for 15 consecutive days. The blood was sampled on Days -1 and 15 for the erythrocyte Pig-a assay and peripheral blood micronucleus assay. Livers were sampled on Day 15 for the liver micronucleus assay and comet assay. Based on the liver micronucleus assay and liver comet assay, AO induced a significant dose-related increase of micronucleated hepatocyte frequencies, and tail DNA percentages, respectively in the middle- and high-dose groups. On the blood micronucleus test and Pig-a assay, no significant increases were observed for the micronucleated reticulocyte frequencies, mutant erythrocyte frequencies (RBCCD59-) or mutant reticulocyte frequencies (RETCD59-) at any of the time points studied. In conclusion, using a multiple-endpoint genotoxicity assay method can reduce the number of experimental animals, boost the efficiency of the experiment, and improve the accuracy of investigations of genotoxicity.

The conjugating green alga Zygnema sp. (Zygnematophyceae) from the Arctic shows high frost tolerance in mature cells (pre-akinetes).

Green algae of the genus Zygnema form extensive mats and produce large amounts of biomass in shallow freshwater habitats. Environmental stresses including freezing may perturb these mats, which usually have only annual character. To estimate the limits of survival at subzero temperatures, freezing resistance of young Zygnema sp. (strain MP2011Skan) cells and pre-akinetes was investigated. Young, 2-week-old cultures were exposed to temperatures of 0 to - 14 °C at 2-K steps, whereas 8-month-old cultures were frozen from - 10 to - 70 °C at 10-K intervals. Cell viability after freezing was determined by 0.1% auramine O vital fluorescence staining and measurements of the effective quantum yield of photosystem II (ФPSII). At - 8 °C, the young vegetative cells were unable to recover from severe frost damage. But temperatures even slightly below zero (- 2 °C) negatively affected the cells' physiology. Single pre-akinetes could survive even at - 70 °C, but their LT50 value was - 26.2 °C. Severe freezing cytorrhysis was observed via cryo-microscopy at - 10 °C, a temperature found to be lethal for young cells. The ultrastructure of young cells appeared unchanged at - 2 °C, but severe damage to biomembranes and formation of small foamy vacuoles was observed at - 10 °C. Pre-akinetes did not show ultrastructural changes at - 20 °C; however, vacuolization increased, and gas bubbles appeared at - 70 °C. Our results demonstrate that the formation of pre-akinetes increases freezing resistance. This adaptation is crucial for surviving the harsh temperature conditions prevailing in the High Arctic in winter and a key feature in seasonal dynamics of Zygnema sp.

Trimellitated sugarcane bagasse: A versatile adsorbent for removal of cationic dyes from aqueous solution. Part II: Batch and continuous adsorption in a bicomponent system.

In the second part of this series of studies, the bicomponent adsorption of safranin-T (ST) and auramine-O (AO) on trimellitated sugarcane bagasse (STA) was evaluated using equimolar dye aqueous solutions at two pH values. Bicomponent batch adsorption was investigated as a function of contact time, solution pH and initial concentration of dyes. Bicomponent kinetic data were fitted by the pseudo-first-order and pseudo-second-order models and the competitive model of Corsel. Bicomponent equilibrium data were fitted by the real adsorbed solution theory model. The antagonistic interactions between ST and AO in the adsorption systems studied contributed to obtain values of maximum adsorption capacity in mono- (Qmax,mono) and bicomponent (Qmax,multi) lower than unity (Qmax,multi/Qmax,mono at pH 4.5 for ST of 0.75 and AO of 0.37 and at pH 7 for ST of 0.94 and AO of 0.43). Mono- and bicomponent adsorption of dyes in a fixed-bed column was evaluated at pH 4.5. The breakthrough curves were fitted by the Thomas and Bohart-Adams original models. Desorption of ST in a fixed-bed column was studied. The results obtained from the bicomponent batch and continuous adsorption showed that the presence of ST most affected the AO adsorption than the presence of AO affected the ST adsorption.