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Experimental Dermatology Sep 2023Wound fluid has been well studied for exploring protein biomarkers contained in it. However, cells in wound fluid have not received much attention due to the difficulty...
Wound fluid has been well studied for exploring protein biomarkers contained in it. However, cells in wound fluid have not received much attention due to the difficulty in their collection. Our study aimed to establish a method for collecting viable cells from discarded wound dressings. A protocol was designed to wash out nonadherent cells and detach adherent cells from silicone-faced foam wound dressings using trypsin-EDTA. The optimal concentration and incubation time of trypsin-EDTA for collecting equivalent proportions of different cell types to the original cell population were determined in vitro. Cell composition and gene expression changes in monocytes, lymphocytes, neutrophils, fibroblasts and keratinocytes were confirmed using immunocytochemistry and RNA-sequencing ex vivo. Full-thickness wounds were created on 9-week-old male C57BL/6J mice. Wound fluid was collected, and half of it was applied to the wound dressings. The original cell population in the wound fluid and the cell population collected from wound dressings were compared. In the in vitro study, 0.25% trypsin-EDTA and 2.5-min incubation time were considered optimal for collecting adherent cells from wound dressings. In the ex vivo study, among all cell types, only CD3+ lymphocytes showed a significantly higher cell proportion in the collected group. The relative gene expression of the five selected cells showed no significant changes (p-value >0.05, |log fold change| < 1.5, differential gene expression analysis). Viable nonadherent and adherent cells were collected from wound dressings without altering gene expression and could be used in future studies for cellular analysis of wound fluid.
Topics: Animals; Mice; Male; Wound Healing; Edetic Acid; Trypsin; Mice, Inbred C57BL; Bandages
PubMed: 37345866
DOI: 10.1111/exd.14857 -
Journal of Affective Disorders Aug 2024Depression, a complex disorder with significant treatment challenges, necessitates innovative therapeutic approaches to address its multifaceted nature and enhance... (Review)
Review
Depression, a complex disorder with significant treatment challenges, necessitates innovative therapeutic approaches to address its multifaceted nature and enhance treatment outcomes. The modulation of KCNQ potassium (K+) channels, pivotal regulators of neuronal excitability and neurotransmitter release, is a promising innovative therapeutic target in psychiatry. Widely expressed across various tissues, including the nervous and cardiovascular systems, KCNQ channels play a crucial role in modulating membrane potential and regulating neuronal activity. Recent preclinical evidence suggests that KCNQ channels, particularly KCNQ3, contribute to the regulation of neuronal excitability within the reward circuitry, offering a potential target for alleviating depressive symptoms, notably anhedonia. Studies using animal models demonstrate that interventions targeting KCNQ channels can restore dopaminergic firing balance and mitigate depressive symptoms. Human studies investigating the effects of KCNQ channel activators, such as ezogabine, have shown promising results in alleviating depressive symptoms and anhedonia. The aforementioned observations underscore the therapeutic potential of KCNQ channel modulation in depression management and highlight the need and justification for phase 2 and phase 3 dose-finding studies as well as studies prespecifying symptomatic targets in depression including anhedonia.
Topics: Humans; Depressive Disorder, Major; KCNQ Potassium Channels; Animals; Phenylenediamines; Carbamates; Anhedonia; KCNQ3 Potassium Channel; Antidepressive Agents
PubMed: 38772507
DOI: 10.1016/j.jad.2024.05.067 -
Chest May 2024Results of retrospective studies have suggested clofazimine as an alternative for rifampicin in the treatment of Mycobacterium avium complex pulmonary disease (MAC-PD). (Randomized Controlled Trial)
Randomized Controlled Trial
BACKGROUND
Results of retrospective studies have suggested clofazimine as an alternative for rifampicin in the treatment of Mycobacterium avium complex pulmonary disease (MAC-PD).
RESEARCH QUESTION
Is a treatment regimen consisting of clofazimine-ethambutol-macrolide noninferior to the standard treatment regimen (rifampicin-ethambutol-macrolide) in the treatment of MAC-PD?
STUDY DESIGN AND METHODS
In this single-center, nonanonymized clinical trial, adult patients with MAC-PD were randomly assigned in a 1:1 ratio to receive rifampicin or clofazimine as adjuncts to an ethambutol-macrolide regimen. The primary outcome was sputum culture conversion following 6 months of treatment.
RESULTS
Forty patients were assigned to receive either rifampicin (n = 19) or clofazimine (n = 21) in addition to ethambutol and a macrolide. Following 6 months of treatment, both arms showed similar percentages of sputum culture conversion based on an intention-to-treat analysis: 58% (11 of 19) for rifampicin and 62% (13 of 21) for clofazimine. Study discontinuation, mainly due to adverse events, was equal in both arms (26% vs 33%). Based on an on-treatment analysis, sputum culture conversion following 6 months of treatment was 79% in both groups. In the clofazimine arm, diarrhea was more prevalent (76% vs 37%; P = .012), while arthralgia was more frequent in the rifampicin arm (37% vs 5%; P = .011). No difference in the frequency of corrected QT interval prolongation was seen between groups.
INTERPRETATION
A clofazimine-ethambutol-macrolide regimen showed similar results to the standard rifampicin-ethambutol-macrolide regimen and should be considered in the treatment of MAC-PD. The frequency of adverse events was similar in both arms, but their nature was different. Individual patient characteristics and possible drug-drug interactions should be taken into consideration when choosing an antibiotic regimen for MAC-PD.
CLINICAL TRIAL REGISTRATION
EudraCT; No.: 2015-003786-28; URL: https://eudract.ema.europa.eu.
Topics: Humans; Rifampin; Clofazimine; Male; Female; Mycobacterium avium-intracellulare Infection; Ethambutol; Drug Therapy, Combination; Middle Aged; Mycobacterium avium Complex; Aged; Treatment Outcome; Antitubercular Agents; Macrolides; Sputum
PubMed: 38040054
DOI: 10.1016/j.chest.2023.11.038 -
Analytical Chemistry Dec 2023Correlating the structure and dynamics of proteins with biological function is critical to understanding normal and dysfunctional cellular mechanisms. We describe a...
Correlating the structure and dynamics of proteins with biological function is critical to understanding normal and dysfunctional cellular mechanisms. We describe a quantitative method of hydroxyl radical generation via Fe(II)-ethylenediaminetetraacetic acid (EDTA)-catalyzed Fenton chemistry that provides ready access to protein oxidative footprinting using equipment commonly found in research and process control laboratories. Robust and reproducible dose-dependent oxidation of protein samples is observed and quantitated by mass spectrometry with as fine a single residue resolution. An oxidation analysis of lysozyme provides a readily accessible benchmark for our method. The efficacy of our oxidation method is demonstrated by mapping the interface of a RAS-monobody complex, the surface of the NIST mAb, and the interface between PRC2 complex components. These studies are executed using standard laboratory tools and a few pennies of reagents; the mass spectrometry analysis can be streamlined to map the protein structure with single amino acid residue resolution.
Topics: Edetic Acid; Hydroxyl Radical; Proteins; Protein Footprinting; Oxidative Stress; Oxidation-Reduction
PubMed: 38049117
DOI: 10.1021/acs.analchem.3c02319 -
Inorganic Chemistry Aug 2023Diamine ligands are effective structural scaffolds for tuning the reactivity of transition-metal complexes for catalytic, materials, and phosphorescent applications and...
Diamine ligands are effective structural scaffolds for tuning the reactivity of transition-metal complexes for catalytic, materials, and phosphorescent applications and have been leveraged for biological use. In this work, we report the synthesis and characterization of a novel class of cyclometalated [C^N] Au(III) complexes bearing secondary diamines including a norbornane backbone, (2,3)-,-dibenzylbicyclo[2.2.1]heptane-2,3-diamine, or a cyclohexane backbone, (1,2)-,-dibenzylcyclohexane-1,2-diamine. X-ray crystallography confirms the square-planar geometry and chirality at nitrogen. The electronic character of the conformationally restricted norbornane backbone influences the electrochemical behavior with redox potentials of -0.8 to -1.1 V, atypical for Au(III) complexes. These compounds demonstrate promising anticancer activity, particularly, complex , which bears a benzylpyridine organogold framework, and supported by the bicyclic conformationally restricted diaminonorbornane, shows good potency in A2780 cells. We further show that a cellular response to evokes reactive oxygen species (ROS) production and does not induce mitochondrial dysfunction. This class of complexes provides significant stability and reactivity for different applications in protein modification, catalysis, and therapeutics.
Topics: Female; Humans; Gold; Antineoplastic Agents; Cell Line, Tumor; Ovarian Neoplasms; Crystallography, X-Ray; Diamines; Norbornanes; Ligands
PubMed: 37530672
DOI: 10.1021/acs.inorgchem.3c02066 -
Australian Endodontic Journal : the... Sep 2023The aim of the study was to determine the properties, efficacy and biocompatibility of combining bromelain enzyme, chlorohexidine and EDTA (BCE) to create a novel...
The aim of the study was to determine the properties, efficacy and biocompatibility of combining bromelain enzyme, chlorohexidine and EDTA (BCE) to create a novel endodontic irrigant. Fourier transform infrared spectrometry was performed to confirm the stability of the BCE and direct contact inhibition test was performed to determine antibacterial action. Baseline pH and surface tension of irrigants was compared with determine stability. Subcutaneous injection to dorsal skin of rabbits was graded for presence of inflammation, oedema, granulation and fibrosis. BCE caused less overall irritation, less oedematous and was earlier to heal than 2.5% NaOCl. The pH stability of BCE was also superior to 2.5% NaOCl. A one-way ANOVA test was performed for the direct contact inhibition and microleakage test. A significant difference was determined (p ≤ 0.05) between BCE and 2.5% NaOCl for antibacterial action. BCE irrigant is effective in preparing dentinal surfaces for root canal without adverse effects and promising longevity.
Topics: Animals; Rabbits; Bromelains; Dental Pulp Cavity; Root Canal Preparation; Root Canal Irrigants; Edetic Acid; Sodium Hypochlorite
PubMed: 36305605
DOI: 10.1111/aej.12704 -
Angewandte Chemie (International Ed. in... May 2024Research on ferroptosis in myocardial ischemia/reperfusion injury (MIRI) using mitochondrial viscosity as a nexus holds great promise for MIRI therapy. However,...
Research on ferroptosis in myocardial ischemia/reperfusion injury (MIRI) using mitochondrial viscosity as a nexus holds great promise for MIRI therapy. However, high-precision visualisation of mitochondrial viscosity remains a formidable task owing to the debilitating electrostatic interactions caused by damaged mitochondrial membrane potential. Herein, we propose a dual-locking mitochondria-targeting strategy that incorporates electrostatic forces and probe-protein molecular docking. Even in damaged mitochondria, stable and precise visualisation of mitochondrial viscosity in triggered and medicated MIRI was achieved owing to the sustained driving forces (e.g., pi-cation, pi-alkyl interactions, etc.) between the developed probe, CBS, and the mitochondrial membrane protein. Moreover, complemented by a western blot, we confirmed that ferrostatin-1 exerts its therapeutic effect on MIRI by improving the system xc/GSH/GPX4 antioxidant system, confirming the therapeutic value of ferroptosis in MIRI. This study presents a novel strategy for developing robust mitochondrial probes, thereby advancing MIRI treatment.
Topics: Ferroptosis; Myocardial Reperfusion Injury; Molecular Docking Simulation; Animals; Mitochondria; Humans; Cyclohexylamines; Phenylenediamines
PubMed: 38509827
DOI: 10.1002/anie.202402537 -
Bioorganic Chemistry Dec 2023New benzimidazole-1,2,3-triazole-quinoline hybrids and their intermediates, differing in substitutions at the C-2 and/or C6 positions of the benzimidazole ring, were...
Synthesis, characterization, in vitro antimycobacterial and cytotoxicity evaluation, DFT calculations, molecular docking and ADME studies of new isomeric benzimidazole-1,2,3-triazole-quinoline hybrid mixtures.
New benzimidazole-1,2,3-triazole-quinoline hybrids and their intermediates, differing in substitutions at the C-2 and/or C6 positions of the benzimidazole ring, were successfully synthesized in 55---80 % yields, with the C6-substituted ones forming as inseparable tautomeric mixtures. The synthesized compounds were fully characterised by FT-IR, 1D- and 2D-NMR, and HRMS. In-depth NMR analysis and DFT molecular calculations showed that the tautomeric mixtures formed in a ratio of almost 1:1 ratio (cis and trans), except for 5 g, where the ratio is 1:2. In vitro antimycobacterial activity evaluation against the H37Rv strain of Mycobacterial tuberculosis was undertaken on all synthesized compounds, and a selected number were further screened for their cytotoxicity on TZM-bl cell lines. Hybrid compounds showed excellent MIC activities ranging from 1.07 to 8.66 μM and were all more efficacious than the first-line reference drug, ethambutol (MIC = 9.54 μM). In particular, hybrid compounds 5b (MIC = 1.54 μM, CC = 58.89 μM and % cell viability = 14.07), 5d (MIC = 2.08 μM, CC = 0.27 μM, and % cell viability = 149.50 %) and 5 g (MIC = 1.49 μM, CC = 4.62 μM and % cell viability = 44.03) were the most promising. Significantly, 5b and 5 g were over six times more efficacious than ethambutol but exhibited cytotoxicity towards TZM-bl cell-lines compared to 5d, which was over four times more active than ethambutol. The physical combination (mimicking combination therapy) of individual pharmacophoric components making up 5 g were less active, indicating the synergistic effect of hybridization. In addition, more than 60 % of all the synthesized hybrids showed better activity than their respective pharmacophoric components. In silico ADME studies of the hybrids revealed favourable physico-chemical properties, while molecular modeling studies suggested binding interactions with Val 61, Gly 62, Glu 65, Ala 66, and Phe 69 amino acid in a reported similar manner to bedaquiline, an approved quinoline-based anti-TB drug.
Topics: Molecular Docking Simulation; Structure-Activity Relationship; Ethambutol; Triazoles; Density Functional Theory; Spectroscopy, Fourier Transform Infrared; Microbial Sensitivity Tests; Anti-Bacterial Agents; Benzimidazoles; Quinolines; Molecular Structure
PubMed: 37832224
DOI: 10.1016/j.bioorg.2023.106904 -
Environmental Research Oct 2023During the mining of rare earth minerals, the application of neodymium-containing manures, and the treatment of spent neodymium iron boron magnet, the generation of...
During the mining of rare earth minerals, the application of neodymium-containing manures, and the treatment of spent neodymium iron boron magnet, the generation of ammonia wastewater containing neodymium is increasing. Thus, the effects of neodymium (Nd(III)) on anaerobic ammonium oxidation (Anammox) were investigated from the aspects of performance, kinetics, statistics, microbial community and sludge morphology, and the recovery strategy of EDTA-2Na wash was discussed. The nitrogen removal efficiency of the Anammox reactor decreased significantly and eventually collapsed at the Nd(III) dosing levels of 20 and 40 mg L, respectively. And the toxicity of Nd(III) to AnAOB was determined by the amount internalized into the cells. The EDTA-2Na wash successfully increased the total nitrogen removal rate (TNRR) of Nd(III)-inhibited Anammox to 41.60% of its initial value within 30 days, and the modified Boltzmann model accurately simulated this recovery process. The transient and extended effects of Nd(III), self-recovery, and EDTA-2Na wash on Anammox were effectively assessed using a one-sample t-test. 16S rRNA gene sequencing indicated that Nd(III) remarkably decreased the relative abundance of Planctomycetes and Candidatus Brocadia. The scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDS) revealed crystal-like neodymium particles on the surface of Anammox sludge. The above-mentioned results demonstrate that the concentration of Nd(III) should be below the toxicity threshold (20 mg L) when treating ammonia wastewater containing neodymium by Anammox, and also emphasize the importance of an appropriate recovery strategy.
Topics: Sewage; Wastewater; Ammonia; Neodymium; RNA, Ribosomal, 16S; Anaerobic Ammonia Oxidation; Edetic Acid; Anaerobiosis; Bioreactors; Oxidation-Reduction; Metals, Rare Earth; Nitrogen; Ammonium Compounds
PubMed: 37467943
DOI: 10.1016/j.envres.2023.116686 -
Journal of Agricultural and Food... Jul 2023Severe plant virus diseases lead to poor harvests and poor crop quality, and the lack of effective suppressive drugs makes plant disease control a huge challenge....
Severe plant virus diseases lead to poor harvests and poor crop quality, and the lack of effective suppressive drugs makes plant disease control a huge challenge. Natural product-based structural simplification is an important strategy for finding novel pesticide candidates. According to our previous research on the antiviral activities of harmine and tetrahydroharmine derivatives, a series of chiral diamine compounds were designed and synthesized by means of structural simplification using diamines in natural products as the core structure in this work, and the antiviral and fungicidal activities were investigated. Most of these compounds displayed higher antiviral activities than those of ribavirin. Compounds and displayed higher antiviral activities than ningnanmycin at 500 μg/mL. The antiviral mechanism research revealed that compounds and could inhibit virus assembly by binding to tobacco mosaic virus (TMV) CP and interfere with the assembly process of TMV CP and RNA transmission electron microscopy and molecular docking. Further fungicidal activity tests showed that these compounds displayed broad-spectrum fungicidal activities. Compounds , , , and with excellent fungicidal activities against f.sp. can be considered as new fungicidal candidates for further research. The current work provides a reference to the development of agricultural active ingredients in crop protection.
Topics: Antiviral Agents; Structure-Activity Relationship; Diamines; Molecular Docking Simulation; Fungicides, Industrial; Tobacco Mosaic Virus; Biological Products; Drug Design
PubMed: 37433073
DOI: 10.1021/acs.jafc.3c01247