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Cytometry. Part a : the Journal of the... Jun 2024Autofluorescence lifetime imaging microscopy (FLIM) is sensitive to metabolic changes in single cells based on changes in the protein-binding activities of the metabolic...
Autofluorescence lifetime imaging microscopy (FLIM) is sensitive to metabolic changes in single cells based on changes in the protein-binding activities of the metabolic co-enzymes NAD(P)H. However, FLIM typically relies on time-correlated single-photon counting (TCSPC) detection electronics on laser-scanning microscopes, which are expensive, low-throughput, and require substantial post-processing time for cell segmentation and analysis. Here, we present a fluorescence lifetime-sensitive flow cytometer that offers the same TCSPC temporal resolution in a flow geometry, with low-cost single-photon excitation sources, a throughput of tens of cells per second, and real-time single-cell analysis. The system uses a 375 nm picosecond-pulsed diode laser operating at 50 MHz, alkali photomultiplier tubes, an FPGA-based time tagger, and can provide real-time phasor-based classification (i.e., gating) of flowing cells. A CMOS camera produces simultaneous brightfield images using far-red illumination. A second PMT provides two-color analysis. Cells are injected into the microfluidic channel using a syringe pump at 2-5 mm/s with nearly 5 ms integration time per cell, resulting in a light dose of 2.65 J/cm that is well below damage thresholds (25 J/cm at 375 nm). Our results show that cells remain viable after measurement, and the system is sensitive to autofluorescence lifetime changes in Jurkat T cells with metabolic perturbation (sodium cyanide), quiescent versus activated (CD3/CD28/CD2) primary human T cells, and quiescent versus activated primary adult mouse neural stem cells, consistent with prior studies using multiphoton FLIM. This TCSPC-based autofluorescence lifetime flow cytometer provides a valuable label-free method for real-time analysis of single-cell function and metabolism with higher throughput than laser-scanning microscopy systems.
PubMed: 38943226
DOI: 10.1002/cyto.a.24883 -
Nitric Oxide : Biology and Chemistry Jun 2024Sodium thiosulfate has been used for decades in the treatment of calciphylaxis and cyanide detoxification, and has recently shown initial therapeutic promise in critical... (Review)
Review
Sodium thiosulfate has been used for decades in the treatment of calciphylaxis and cyanide detoxification, and has recently shown initial therapeutic promise in critical diseases such as neuronal ischemia, diabetes mellitus, heart failure and acute lung injury. However, the precise mechanism of sodium thiosulfate remains incompletely defined and sometimes contradictory. Although sodium thiosulfate has been widely accepted as a donor of hydrogen sulfide (HS), emerging findings suggest that it is the executive signaling molecule for HS and that its effects may not be dependent on HS. This article presents an overview of the current understanding of sodium thiosulfate, including its synthesis, biological characteristics, and clinical applications of sodium thiosulfate, as well as the underlying mechanisms in vivo. We also discussed the interplay of sodium thiosulfate and HS. Our review highlights sodium thiosulfate as a key player in sulfide signaling with the broad clinical potential for the future.
PubMed: 38897561
DOI: 10.1016/j.niox.2024.06.004 -
Evaluation of gold adsorption on activated carbon from real cyanide and thiourea leachate solutions.Heliyon Jun 2024The recovery of gold by adsorption using activated carbon from sodium cyanide and thiourea leached solutions are reported in this study. The leached solutions were...
The recovery of gold by adsorption using activated carbon from sodium cyanide and thiourea leached solutions are reported in this study. The leached solutions were obtained under real operating conditions from the beneficiation plant "Paz Borja", Machala-Ecuador. Calgon Carbon DG-11 6X12 type, widely used in the local metallurgical industry was used as adsorbent material. The operational parameters varied during the adsorption process experiments included the concentration of leaching agent, agitation speed, dose of activated carbon and initial concentration of gold. The control parameters included density, percentage of solid, pH, temperature, and solution potential. The obtained results were adjusted to mass transfer model by diffusion through the interface and the Freundlich model for the equilibrium isotherms. The analysis of the results indicates a higher adsorption rate of the gold di-cyanide complex on activated carbon compared to gold-thiourea complexes.
PubMed: 38841478
DOI: 10.1016/j.heliyon.2024.e31606 -
BioRxiv : the Preprint Server For... May 2024Autofluorescence lifetime imaging microscopy (FLIM) is sensitive to metabolic changes in single cells based on changes in the protein-binding activities of the metabolic...
Autofluorescence lifetime imaging microscopy (FLIM) is sensitive to metabolic changes in single cells based on changes in the protein-binding activities of the metabolic co-enzymes NAD(P)H. However, FLIM typically relies on time-correlated single-photon counting (TCSPC) detection electronics on laser-scanning microscopes, which are expensive, low-throughput, and require substantial post-processing time for cell segmentation and analysis. Here, we present a fluorescence lifetime-sensitive flow cytometer that offers the same TCSPC temporal resolution in a flow geometry, with low-cost single-photon excitation sources, a throughput of tens of cells per second, and real-time single-cell analysis. The system uses a 375nm picosecond-pulsed diode laser operating at 50MHz, alkali photomultiplier tubes, an FPGA-based time tagger, and can provide real-time phasor-based classification ( ., gating) of flowing cells. A CMOS camera produces simultaneous brightfield images using far-red illumination. A second PMT provides two-color analysis. Cells are injected into the microfluidic channel using a syringe pump at 2-5 mm/s with nearly 5ms integration time per cell, resulting in a light dose of 2.65 J/cm that is well below damage thresholds (25 J/cm at 375 nm). Our results show that cells remain viable after measurement, and the system is sensitive to autofluorescence lifetime changes in Jurkat T cells with metabolic perturbation (sodium cyanide), quiescent vs. activated (CD3/CD28/CD2) primary human T cells, and quiescent vs. activated primary adult mouse neural stem cells, consistent with prior studies using multiphoton FLIM. This TCSPC-based autofluorescence lifetime flow cytometer provides a valuable label-free method for real-time analysis of single-cell function and metabolism with higher throughput than laser-scanning microscopy systems.
PubMed: 38798331
DOI: 10.1101/2024.05.15.594394 -
International Journal of... May 2024Pakistan is an agricultural country producing plenty of fruits, like: mango, banana, apple, peaches, grapes, plums, variety of citrus fruits including lemon, grapefruit,...
Pakistan is an agricultural country producing plenty of fruits, like: mango, banana, apple, peaches, grapes, plums, variety of citrus fruits including lemon, grapefruit, and oranges. So far the peels of most of the fruits are usually wasted and not properly utilized anywhere. In this work, the peels of banana and grapefruit are converted into biochar by slow pyrolysis under controlled supply of air and used for sequestering cyanide ions from aqueous medium after chemical modification with ZnCl and sodium dodecyl sulfate (SDS). The modified biochar was characterized by various instrumental techniques, like: SEM, FTIR, TGA, and CHNS. Different parameters, like: time, temperature, pH, and dose of adsorbent affecting the adsorption of cyanide ions, onto prepared biochar were optimized and to understand the adsorption phenomenon, kinetic and thermodynamic studies were performed. Concentration of cyanide ions was estimated by employing standard ion selective electrode system and it is found that Sodium Dodecyl Sulfate treated biochar of banana peels shown more adsorption capacity, : 17.080 mg/g as compared to all samples. Present work revealed that the biochar produced from the fruit waste has sufficient potential to eliminate trace quantities of cyanide from water, especially after treatment with sodium dodecyl sulfate.
PubMed: 38755758
DOI: 10.1080/15226514.2024.2349721 -
Highly Efficient Recovery of Au(I) from Gold Leaching Solution Using Sodium Dimethyldithiocarbamate.ACS Omega May 2024As a sustainable, nontoxic and environmentally friendly cyanide-free gold leaching agent, thiosulfate has been applied to some extent in the field of hydrometallurgy....
As a sustainable, nontoxic and environmentally friendly cyanide-free gold leaching agent, thiosulfate has been applied to some extent in the field of hydrometallurgy. However, the difficult recovery of gold ions in gold leaching solutions limits further application of thiosulfate gold leaching technology. This study demonstrated the feasibility of gold recovery by sodium dimethyldithiocarbamate (SDD) precipitation and recycling of ammonia and a lixiviant in solution. SDD achieved the purpose of recovering gold by forming granular precipitates with gold ions in solution. It can almost completely recover gold ions in 2.5-17.34 mg/L of gold leaching solution within 1 min at 25 °C, in which a gold recovery capacity of 7.99 kg/t is achieved. The leaching rate of gold ore did not change significantly after recycling the residual ammonia and thiosulfate in the leaching solution after gold recovery by SDD, and its leaching rate basically remained at 81%. The mechanism of SDD recovering Au was determined to involve the ligand exchange of SDD and Au[(SO)]. Moreover, the interaction mechanism between SDD and Au(I) was further validated by density functional theory calculations. Considering its low cost, simple technology, and environmental friendliness, the SDD precipitation process has the potential for large-scale application in gold recovery from thiosulfate gold leaching solutions.
PubMed: 38737059
DOI: 10.1021/acsomega.4c01941 -
Molecules (Basel, Switzerland) May 2024We modified -symmetric benzene-1,3,5--amide (BTA) by introducing flexible linkers in order to generate an N-centered BTA (N-BTA) molecule. The N-BTA compound formed gels...
We modified -symmetric benzene-1,3,5--amide (BTA) by introducing flexible linkers in order to generate an N-centered BTA (N-BTA) molecule. The N-BTA compound formed gels in alcohols and aqueous mixtures of high-polar solvents. Rheological studies showed that the DMSO/water (1:1, /) gels were mechanically stronger compared to other gels, and a similar trend was observed for thermal stability. Powder X-ray analysis of the xerogel obtained from various aqueous gels revealed that the packing modes of the gelators in these systems were similar. The stimuli-responsive properties of the N-BTA towards sodium/potassium salts indicated that the gel network collapsed in the presence of more nucleophilic anions such as cyanide, fluoride, and chloride salts at the MGC, but the gel network was intact when in contact with nitrate, sulphate, acetate, bromide, and iodide salts, indicating the anion-responsive properties of N-BTA gels. Anion-induced gel formation was observed for less nucleophilic anions below the MGC of N-BTA. The ability of N-BTA gels to act as an adsorbent for hazardous anionic and cationic dyes in water was evaluated. The results indicated that the ethanolic gels of N-BTA successfully absorbed methylene blue and methyl orange dyes from water. This work demonstrates the potential of the N-BTA gelator to act as a stimuli-responsive material and a promising candidate for water purification.
PubMed: 38731640
DOI: 10.3390/molecules29092149 -
Biotechnology Advances 2024Indigo is a natural dye extensively used in the global textile industry. However, the conventional synthesis of indigo using toxic compounds like aniline, formaldehyde,... (Review)
Review
Indigo is a natural dye extensively used in the global textile industry. However, the conventional synthesis of indigo using toxic compounds like aniline, formaldehyde, and hydrogen cyanide has led to environmental pollution and health risks for workers. This method also faces growing economic, sustainability, and environmental challenges. To address these issues, the concept of bio-indigo or indigo biosynthesis has been proposed as an alternative to aniline-based indigo synthesis. Among various enzymes, Flavin-containing Monooxygenases (FMOs) have shown promise in achieving a high yield of bio-indigo. However, the industrialization of indigo biosynthesis still encounters several challenges. This review focuses on the historical development of indigo biosynthesis mediated by FMOs. It highlights several factors that have hindered industrialization, including the use of unsuitable chassis (Escherichia coli), the toxicity of indole, the high cost of the substrate L-tryptophan, the water-insolubility of the product indigo, the requirement of reducing reagents such as sodium dithionite, and the relatively low yield and high cost compared to chemical synthesis. Additionally, this paper summarizes various strategies to enhance the yield of indigo synthesized by FMOs, including redundant sequence deletion, semi-rational design, cheap precursor research, NADPH regeneration, large-scale fermentation, and enhancement of water solubility of indigo.
Topics: Indigo Carmine; Mixed Function Oxygenases; Oxygenases; Coloring Agents; Escherichia coli
PubMed: 38729229
DOI: 10.1016/j.biotechadv.2024.108374 -
Animals : An Open Access Journal From... Feb 2024plants naturally produce dhurrin, a cyanogenic glycoside that may be hydrolysed to cyanide, resulting in often-lethal toxicoses. Ruminants are particularly sensitive to...
plants naturally produce dhurrin, a cyanogenic glycoside that may be hydrolysed to cyanide, resulting in often-lethal toxicoses. Ruminants are particularly sensitive to cyanogenic glycosides due to the active role of rumen microbiota in dhurrin hydrolysis. This work provides an overview of a poisoning outbreak that occurred in 5 farms in Northwest Italy in August 2022; a total of 66 cows died, and many others developed acute toxicosis after being fed on either cultivated () or wild Sorghum (). Clinical signs were recorded, and all cows received antidotal/supportive therapy. Dead animals were subjected to necropsy, and dhurrin content was determined in Sorghum specimens using an LC-MS/MS method. Rapid onset, severe respiratory distress, recumbency and convulsions were the main clinical features; bright red blood, a bitter almond smell and lung emphysema were consistently observed on necropsy. The combined i.v. and oral administration of sodium thiosulphate resulted in a rapid improvement of clinical signs. Dhurrin concentrations corresponding to cyanide levels higher than the tolerated threshold of 200 mg/kg were detected in sorghum specimens from 4 out of 5 involved farms; thereafter, such levels declined, reaching tolerable concentrations in September-October. Feeding cattle with wild or cultivated Sorghum as green fodder is a common practice in Northern Italy, especially in summer. However, care should be taken in case of adverse climatic conditions, such as severe drought and tropical temperatures (characterising summer 2022), which are reported to increase dhurrin synthesis and storage.
PubMed: 38473128
DOI: 10.3390/ani14050743 -
Current Microbiology Feb 2024Uncouplers of oxidative phosphorylation dissipate the proton gradient, causing lower ATP production. Bacteria encounter several non-classical uncouplers in the...
Uncouplers of oxidative phosphorylation dissipate the proton gradient, causing lower ATP production. Bacteria encounter several non-classical uncouplers in the environment, leading to stress-induced adaptations. Here, we addressed the molecular mechanisms responsible for the effects of uncouplers in Escherichia coli. The expression and functions of genes involved in phenotypic antibiotic resistance were studied using three compounds: two strong uncouplers, i.e., Carbonyl cyanide m-chlorophenyl hydrazone (CCCP) and 2,4-Dinitrophenol (DNP), and one moderate uncoupler, i.e., Sodium salicylate (NaSal). Quantitative expression studies demonstrated induction of transcripts encoding marA, soxS and acrB with NaSal and DNP, but not CCCP. Since MarA and SoxS are degraded by the Lon protease, we investigated the roles of Lon using a lon-deficient strain (Δlon). Compared to the wild-type strain, Δlon shows compromised growth upon exposure to NaSal or 2, 4-DNP. This sensitivity is dependent on marA but not rob and soxS. On the other hand, the Δlon strain shows enhanced growth in the presence of CCCP, which is dependent on acrB. Interestingly, NaSal and 2,4-DNP, but not CCCP, induce resistance to antibiotics, such as ciprofloxacin and tetracycline. This study addresses the effects of uncouplers and the roles of genes involved during bacterial growth and phenotypic antibiotic resistance. Strong uncouplers are often used to treat wastewater, and these results shed light on the possible mechanisms by which bacteria respond to uncouplers. Also, the rampant usage of some uncouplers to treat wastewater may lead to the development of antibiotic resistance.
Topics: Escherichia coli; Oxidative Phosphorylation; Protease La; Carbonyl Cyanide m-Chlorophenyl Hydrazone; Wastewater; Anti-Bacterial Agents; Dinitrophenols; Escherichia coli Proteins
PubMed: 38372817
DOI: 10.1007/s00284-024-03632-w