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Hematin plays a crucial role in various physiological functions, and the determination of hematin in complex biological matrixes is a significant but difficult issue. Considering the unique photophysical/photochemical properties of carbon quantum dots (CQDs) prepared with p-aminobenzoic acid (PABA) and ethanol, a new strategy for the design of fluorescent probes for hematin has been achieved. The proposed sensor array is fabricated based on the inner filter effect (IFE) between hematin and CQDs with phenomenon of selective fluorescence quenching of CQDs which results from the strong absorption of the excitation and emission spectrum of CQDs by hematin. The fluorescence quenching of CQDs is closely related to the amount of hematin and there is a good linear relationship over the range of 0.5-10μM with a detection limit of 0.25μM. What's more, the fluorescence assay has been successfully applied for hematin sensing in healthy human red cells showing this sensing assay has a great potential prospect for detection of hematin in the complex matrixes., (Copyright © 2017 Elsevier B.V. All rights reserved.)

During malarial anemia, 20 uninfected red blood cells (RBCs) are destroyed for every RBC infected by Plasmodium falciparum (Pf). Increasing evidence indicates an important role for complement in destruction of uninfected RBCs. Products of RBC lysis induced by Pf, including the digestive vacuole and hematin, activate complement and promote C3 fragment deposition on uninfected RBCs. C3-opsonized cells are then subject to extravascular destruction mediated by fixed tissue macrophages which express receptors for C3 fragments. The Compstatin family of cyclic peptides blocks complement activation at the C3 cleavage step, and is under investigation for treatment of complement-mediated diseases. We demonstrate, that under a variety of stringent conditions, second-generation Compstatin analogue Cp40 completely blocks hematin-mediated deposition of C3 fragments on naïve RBCs. Our findings indicate that prophylactic provision of Compstatin for malaria-infected individuals at increased risk for anemia may provide a safe and inexpensive treatment to prevent or substantially reduce malarial anemia., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Pathologies such as malaria, hemorrhagic stroke, sickle cell disease, and thalassemia are characterized by the release of hemoglobin degradation products from damaged RBCs. Hematin (liganded with OH−) and hemin (liganded with Cl−)—are the oxidized forms of heme with toxic properties due to their hydrophobicity and the presence of redox-active Fe3. In the present study, using the original LaSca-TM laser particle analyzer, flow cytometry, and confocal microscopy, we showed that both hematin and hemin induce dose-dependent RBC spherization and hemolysis with ghost formation. Hematin and hemin at nanomolar concentrations increased [Ca2+]i in RBC; however, spherization and hemolysis occurred in the presence and absence of calcium, indicating that both processes are independent of [Ca2+]i. Both compounds triggered acute phosphatidylserine exposure on the membrane surface, reversible after 60 min of incubation. A comparison of hematin and hemin effects on RBCs revealed that hematin is a more reactive toxic metabolite than hemin towards human RBCs. The toxic effects of heme derivatives were reduced and even reversed in the presence of albumin, indicating the presence in RBCs of the own recovery system against the toxic effects of heme derivatives. [ABSTRACT FROM AUTHOR]

Pathologies such as malaria, hemorrhagic stroke, sickle cell disease, and thalassemia are characterized by the release of hemoglobin degradation products from damaged RBCs. Hematin (liganded with OH−) and hemin (liganded with Cl−)—are the oxidized forms of heme with toxic properties due to their hydrophobicity and the presence of redox-active Fe3. In the present study, using the original LaSca-TM laser particle analyzer, flow cytometry, and confocal microscopy, we showed that both hematin and hemin induce dose-dependent RBC spherization and hemolysis with ghost formation. Hematin and hemin at nanomolar concentrations increased [Ca2+]i in RBC; however, spherization and hemolysis occurred in the presence and absence of calcium, indicating that both processes are independent of [Ca2+]i. Both compounds triggered acute phosphatidylserine exposure on the membrane surface, reversible after 60 min of incubation. A comparison of hematin and hemin effects on RBCs revealed that hematin is a more reactive toxic metabolite than hemin towards human RBCs. The toxic effects of heme derivatives were reduced and even reversed in the presence of albumin, indicating the presence in RBCs of the own recovery system against the toxic effects of heme derivatives.

Pooled Immunoglobulin G (IgG), hematin and the membrane-disruptive amphipathic peptide melittin have received attention as powerful biomacromolecules for biomedical and pharmacology applications. Their action on surface properties, oxidation status and epifluorescence properties measured in vitro provide useful information about the functional activity of upper biomacromolecules in erythrocytes in vivo. The hemolysis of erythrocyte membranes, as well as changes in hematocrit and the morphology of erythrocytes, was investigated here via fluorescence microscopy using FITC-concanavalin A binding to cells. The effect of melittin on the membrane capacitance and resistance of model lipid bilayers was probed via electrochemical impedance spectroscopy. Lipid bilayer capacitance was higher in the presence of 0.10 g/L melittin compared to that in the control, which is likely related to bilayer thinning and alterations of the dielectric permittivity of melittin-treated membranes. The biomolecule interactions with red blood cells were probed in physiological media in which the surface of erythrocyte membranes was negatively charged. Surface parameters of erythrocytes are reported upon IgG/hematin and IgG/melittin treatment. Pooled IgG in the presence of melittin, preincubated IgG/hematin preparations promoted a significant decrease in the electrokinetic potential of erythrocytes (Rh-positive). A malondialdehyde (MDA) assay revealed a high rate of lipid peroxidation in erythrocytes treated with IgG/hematin or IgG/melittin preparations. This finding might be a result of pooled IgG interactions with the hematin molecule and the subsequent conformational changes in the protein molecule altering the electrokinetic properties of the erythrocyte membrane related to the Rh group type of erythrocytes. The pooled IgG and hematin are reported to have important consequences for the biophysical understanding of the immunopathological mechanisms of inflammatory, autoimmune and antibody-mediated pathological processes.

Nitrogen-doped MoS 2 quantum dots (N-MoS 2 QDs) were synthesized via a facile hydrothermal approach, and exhibited high fluorescence quantum yield (QY, 14.9%), excellent photostability, biocompatibility and water solubility. A novel method with good selectivity and sensitivity was established to assay hematin using N-MoS 2 QDs as a fluorescent probe based on inner filter effect (IFE). Fluorescent quenching of N-MoS 2 QDs has a fine linear dependence with the concentration of hematin in the range of 0.5–15 μmol/L and a limit of detection of 0.32 μmol/L (S/N = 3). By the detection method, average concentration of hematin in real health human erythrocytes was measured as 22.5 ± 3.9 μmol/L. And, recoveries range varied from 94 to 108% through standard recovery experiment. The N-MoS 2 QDs probe shows excellent photostability, low cytotoxicity and anti-interference ability for hematin assay, which may become a promising method for the test of hematin in human blood. Unlabelled Image • Ultrathin nitrogen-doped MoS 2 QDs was successfully synthesized by a facile "one-pot" hydrothermal method • As-prepared N-MoS 2 QDs exhibit excellent tunable photoluminescence properties • N-MoS 2 QDs has good water solubility, low biotoxicity, high photostability and biocompatibility • The N-MoS 2 QDs based fluorescent sensor is capable of assay for free hematin in complex biological matrixes [ABSTRACT FROM AUTHOR]