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Heliyon May 2024Tracing individual cell pathways among the whole population is crucial for understanding their behavior, cell communication, migration dynamics, and fate. Optical...
Tracing individual cell pathways among the whole population is crucial for understanding their behavior, cell communication, migration dynamics, and fate. Optical labeling is one approach for tracing individual cells, but it typically requires genetic modification to induce the generation of photoconvertible proteins. Nevertheless, this approach has limitations and is not applicable to certain cell types. For instance, genetic modification often leads to the death of macrophages. This study aims to develop an alternative method for labeling macrophages by utilizing photoconvertible micron-sized capsules capable of easy internalization and prolonged retention within cells. Thermal treatment in a polyvinyl alcohol gel medium is employed for the scalable synthesis of capsules with a wide range of fluorescent dyes, including rhodamine 6G, pyronin B, fluorescein, acridine yellow, acridine orange, thiazine red, and previously reported rhodamine B. The fluorescence brightness, photostability, and photoconversion ability of the capsules are evaluated using confocal laser scanning microscopy. Viability, uptake, mobility, and photoconversion studies are conducted on RAW 264.7 and bone marrow-derived macrophages, serving as model cell lines. The production yield of the capsules is increased due to the use of polyvinyl alcohol gel, eliminating the need for conventional filtration steps. Capsules entrapping rhodamine B and rhodamine 6G meet all requirements for intracellular use in individual cell tracking. Mass spectrometry analysis reveals a sequence of deethylation steps that result in blue shifts in the dye spectra upon irradiation. Cellular studies on macrophages demonstrate robust uptake of the capsules. The capsules exhibit minimal cytotoxicity and have a negligible impact on cell motility. The successful photoconversion of RhB-containing capsules within cells highlights their potential as alternatives to photoconvertible proteins for individual cell labeling, with promising applications in personalized medicine.
PubMed: 38813172
DOI: 10.1016/j.heliyon.2024.e30680 -
RSC Advances Feb 2024Applications of 9-aminoacridine (9aa) and its derivatives span fields such as chemistry, biology, and medicine, including anticancer and antimicrobial activities....
Applications of 9-aminoacridine (9aa) and its derivatives span fields such as chemistry, biology, and medicine, including anticancer and antimicrobial activities. Protonation of such molecules can alter their bioavailability as weakly basic drugs like aminoacridines exhibit reduced solubility at high pH levels potentially limiting their effectiveness in patients with elevated gastric pH. In this study, we analyse the influence of protonation on the electronic characteristics of the molecular organic crystals of 9-aminoacridine. The application of quantum crystallography, including aspherical atom refinement, has enriched the depiction of electron density in the studied systems and non-covalent interactions, providing more details than previous studies. Our experimental results, combined with a topological analysis of the electron density and its Laplacian, provided detailed descriptions of how protonation changes the electron density distribution around the amine group and water molecule, concurrently decreasing the electron density at bond critical points of N/O-H bonds. Protonation also alters the molecular architecture of the systems under investigation. This is reflected in different proportions of the N⋯H and O⋯H intermolecular contacts for the neutral and protonated forms. Periodic DFT calculations of the cohesive energies of the crystal lattice, as well as computed interaction energies between molecules in the crystal, confirm that protonation stabilises the crystal structure due to a positive synergy between strong halogen and hydrogen bonds. Our findings highlight the potential of quantum crystallography in predicting crystal structure properties and point to its possible applications in developing new formulations for poorly soluble drugs.
PubMed: 38348299
DOI: 10.1039/d3ra08081a -
Scientific Reports Apr 2024Efficient separation of electron-hole pairs remains pivotal in optimizing photogenerated carrier functionality across diverse catalytic and optoelectronic systems. This...
Efficient separation of electron-hole pairs remains pivotal in optimizing photogenerated carrier functionality across diverse catalytic and optoelectronic systems. This study presents the fabrication of a novel hollow direct Z-scheme photocatalyst, ZnO/TiO. A thorough analysis encompassing various techniques such as Ultraviolet-Visible Spectroscopy (UV-Vis), X-ray Diffraction (XRD), Transmission electron Microscopy (TEM), Fourier Transform Infrared Spectroscopy (FT-IR), Thermogravimetric Analysis (TGA), and Energy-Dispersive X-ray Spectroscopy (EDX) provided detailed insights into the complex material characteristics of the ZnO/TiO heterojunction catalyst. The findings revealed coexisting anatase TiO and wurtzite ZnO phases, each retaining distinct attributes within the nanocomposites (NCs) structure. The study showcased the photocatalytic efficacy of ZnO/TiO-NCs in decomposing Methylene Blue and Acridine Orange under UV irradiation, correlated with their underlying structures. Enhanced degradation of these dyes resulted from the establishment of a direct Z-scheme heterojunction between ZnO and TiO. Employing Density Functional Theory (DFT) using Quantum ESPRESSO, this research analyzed phase diagrams and band structures, elucidating electronic properties and structural correlations. The study characterized a ZnO/TiO composite, revealing a band gap of 3.1-3.3 eV through UV-Visible spectroscopy and confirming its formation without impurity phases via XRD analysis. TEM and EDX showed uniform element dispersion (Zn: 27%, Ti: 29.62%, C: 5.03%, O: 38.35%). Computational analysis using DFT indicated a reduction in stable phases with increasing temperature. Enhanced dye degradation was observed (MB: 88.9%, AO: 84%), alongside significant antibacterial activity. In the future we predict that research will focus on development of scaled up production and photocatalytic activity through surface modification, while unveiling mechanistic insights and environmental applicability for multifunctional use in water treatment and antibacterial applications, leading to further advancement of the field.
PubMed: 38575610
DOI: 10.1038/s41598-024-57392-5 -
Asian Journal of Andrology Oct 2023Aberrant sperm protamination is linked to sperm dysmorphology and nuclear chromatin condensation. Yet, its effects on sperm cytoplasmic maturation remain largely...
Aberrant sperm protamination is linked to sperm dysmorphology and nuclear chromatin condensation. Yet, its effects on sperm cytoplasmic maturation remain largely unexplored. The relationships of protamines, sperm morphology, DNA damage, and cytoplasmic remodeling were illustrated in this study to provide fresh perspectives on the mechanisms of male infertility. A total of 205 infertile males were allocated into 5 groups according to the percentage of spermatozoa exhibiting abnormal morphology within their samples. Sperm concentration, motility, abnormal sperm morphology, cytoplasmic droplets (CDs), and excess residual cytoplasm (ERC) were analyzed according to the World Health Organization manual (2010). Sperm nuclear vacuoles (NVs) were determined by propidium iodide (PI) staining. Sperm protamine expressions (P1 and P2) were detected by western blot. DNA damage was measured by acridine orange test (AOT) to calculate the proportion of sperm with single-strand DNA breaks (SSBs). Our data showed that sperm concentration and motility in infertile males significantly decreased with the severity of abnormal sperm morphology (both P < 0.01). P1 level, P1/P2 ratio, and SSB rate increased with the severity of sperm dysmorphology, whilst the P2 level decreased (all P < 0.01). NVs, CDs, and ERC were more common in males with sperm dysmorphology and positively correlated with the SSB rate (all P < 0.01). The relationships between the SSB rate and the P1/P2 ratio were also significant (P < 0.01). Aberrant protamination may cause sperm dysmorphology and compromise male fertility by impairing sperm's nucleus and cytoplasm maturation, with the P1/P2 ratio potentially serving as a valuable indicator of sperm quality and male fertility.
PubMed: 37921517
DOI: 10.4103/aja202360 -
Nanoscale Advances Oct 2023A phenoxyaniline-based macroinitiator is utilized for the first time in order to produce phenoxyaniline--poly(methyl methacrylate) composites through single electron...
A phenoxyaniline-based macroinitiator is utilized for the first time in order to produce phenoxyaniline--poly(methyl methacrylate) composites through single electron transfer-living radical polymerization (SET-LRP) under mild conditions. A different weight percentage of Cloisite 93A is added into the polymer mixtures in order to increase their biochemical properties. The prepared block copolymer nanocomposites are characterized using ATR-IR, UV-vis-spectroscopy, XRD, Raman, TGA, DSC, a particle size analyzer, contact angle measurements and SEM in order to characterize their structural, thermal, surface and morphological properties. Further, the developed polymeric nanocomposites are successfully applied in two different cancer cell lines (prostate adenocarcinoma and lung cancer), which show excellent anticancer properties. Also, acridine orange/ethidium bromide (AO/EtBr) dual staining is performed, which causes drastic cell death by apoptosis in both A549 and PC-3 cell lines, which indicated that the prepared polymeric nanocomposites effectively inhibit the cell proliferation and induce the apoptosis in both the cancer cells. Here nanoclay is used for cancer treatment because of its complete water solubility, which essentially causes the formation of a cationic complex between the clay and drug through electrostatic interactions. Hence, the exchange of ions between the clay and other ions in the biological environment leads to inhibition of the proliferation of prostate adenocarcinoma and lung cancer cells in the system.
PubMed: 37881709
DOI: 10.1039/d3na00644a -
Cells Mar 2024Macroautophagy (hereafter autophagy) is a lysosomal degradation pathway that functions in nutrient recycling and as a mechanism of innate immunity. Previously, we...
Macroautophagy (hereafter autophagy) is a lysosomal degradation pathway that functions in nutrient recycling and as a mechanism of innate immunity. Previously, we reported a novel host-bacteria interaction between cariogenic and bitter taste receptor (T2R14) in gingival epithelial cells (GECs), leading to an innate immune response. Further, might be using the host immune system to inhibit other Gram-positive bacteria, such as . To determine whether these bacteria exploit the autophagic machinery of GEC, it is first necessary to evaluate the role of T2R14 in modulating autophagic flux. So far, the role of T2R14 in the regulation of autophagy is not well characterized. Therefore, in this study, for the first time, we report that T2R14 downregulates autophagy flux in GECs, and T2R14 knockout increases acidic vacuoles. However, the treatments of GEC WT with a T2R14 agonist and antagonist did not lead to a significant change in acidic vacuole formation. Transmission electron microscopy morphometric results also suggested an increased number of autophagic vesicles in T2R14-knockout GEC. Further, our results suggest that competence stimulating peptide CSP-1 showed robust intracellular calcium release and this effect is both T2R14- and autophagy protein 7-dependent. In this study, we provide the first evidence that T2R14 modulates autophagy flux in GEC. The results of the current study could help in identifying the impact of T2R in regulation of the immuno-microenvironment of GEC and subsequently oral health.
Topics: Taste; Receptors, G-Protein-Coupled; Staphylococcus aureus; Autophagy; Epithelial Cells
PubMed: 38534375
DOI: 10.3390/cells13060531 -
Frontiers in Chemistry 2023Despite modern sciences and advancements in new drugs or chemicals, the new era now rushes natural remedies for various illnesses and diseases that lead to end organ...
Despite modern sciences and advancements in new drugs or chemicals, the new era now rushes natural remedies for various illnesses and diseases that lead to end organ damage. In this study, we investigated ethanolic extract's effect against doxorubicin-induced cardiotoxicity and renal toxicity. To determine phytochemicals, a phytochemical screening was conducted. Various assays were used to measure the antioxidant activity, including the DPPH (2,2-diphenylpicrylhydrazyl), SOD (superoxide dismutase), NO (nitric oxide), and others. The antiproliferative effect of was assessed by MTT assay; morphological analysis was performed using an inverted and phase contrast microscope, ultra morphological analysis of apoptosis with acridine orange (AO)/propidium iodide (PI) staining. It was seen that doxorubicin caused elevated serum markers and abnormal changes in histological patterns. The significant reduction in cardiac and renal marker levels seen in groups given either 400 or 600 mg/kg of crude extract demonstrates that has a protective effect against doxorubicin-induced cardiotoxicity due to the presence of active phytoconstituents having antioxidant potential. There is a dose-dependent decrease in cell viability when using . Apoptosis was observed in the treated cells. In conclusion, our research lends credence to the idea that could be used for cancer management and have cardioprotective and nephroprotective effects.
PubMed: 38164252
DOI: 10.3389/fchem.2023.1283618 -
Animals : An Open Access Journal From... Jul 2023This study aimed to analyze various alterations in the morphology of the sperm head and its association with nucleus instability and insufficient sperm protamine....
This study aimed to analyze various alterations in the morphology of the sperm head and its association with nucleus instability and insufficient sperm protamine. Frozen-thawed semen from twenty local Indonesian bulls was used for all stages in this study. The results of sperm head defect assessments are used for bull grouping, high (HD) and low (LD). Sperm DNA damage was assessed using Acridine Orange and Halomax. The PRM1 protein abundance was carried out using an enzyme immunoassay, while PRM1 gene expression was carried out using the RT-qPCR. PRM deficiency was performed using CMA. Several kinds of sperm head defects in the HD were significantly higher ( < 0.05) than in the LD bulls. Sperm DNA damage showed a significant ( < 0.05) difference between the HD and LD bulls. PRM1 abundance was significantly ( < 0.05) decreased in HD bulls. PRM deficiency was significantly ( < 0.05) higher in HD bulls than in LD bulls. PRM deficiency in bulls correlated significantly ( < 0.01) with sperm head defects, DNA damage, and PRM1 abundance. The lack of sperm protamine might affect the sperm nucleus's stability and induce morphological alterations in the sperm head.
PubMed: 37570242
DOI: 10.3390/ani13152433 -
Trials Dec 2023Red blood cell (RBC) transfusion is a critical supportive therapy in cardiovascular surgery (CVS). Donor selection and testing have reduced the risk of...
Evaluation of the efficacy and safety of amustaline/glutathione pathogen-reduced RBCs in complex cardiac surgery: the Red Cell Pathogen Inactivation (ReCePI) study-protocol for a phase 3, randomized, controlled trial.
BACKGROUND
Red blood cell (RBC) transfusion is a critical supportive therapy in cardiovascular surgery (CVS). Donor selection and testing have reduced the risk of transfusion-transmitted infections; however, risks remain from bacteria, emerging viruses, pathogens for which testing is not performed and from residual donor leukocytes. Amustaline (S-303)/glutathione (GSH) treatment pathogen reduction technology is designed to inactivate a broad spectrum of infectious agents and leukocytes in RBC concentrates. The ReCePI study is a Phase 3 clinical trial designed to evaluate the efficacy and safety of pathogen-reduced RBCs transfused for acute anemia in CVS compared to conventional RBCs, and to assess the clinical significance of treatment-emergent RBC antibodies.
METHODS
ReCePI is a prospective, multicenter, randomized, double-blinded, active-controlled, parallel-design, non-inferiority study. Eligible subjects will be randomized up to 7 days before surgery to receive either leukoreduced Test (pathogen reduced) or Control (conventional) RBCs from surgery up to day 7 post-surgery. The primary efficacy endpoint is the proportion of patients transfused with at least one study transfusion with an acute kidney injury (AKI) diagnosis defined as any increased serum creatinine (sCr) level ≥ 0.3 mg/dL (or 26.5 µmol/L) from pre-surgery baseline within 48 ± 4 h of the end of surgery. The primary safety endpoints are the proportion of patients with any treatment-emergent adverse events (TEAEs) related to study RBC transfusion through 28 days, and the proportion of patients with treatment-emergent antibodies with confirmed specificity to pathogen-reduced RBCs through 75 days after the last study transfusion. With ≥ 292 evaluable, transfused patients (> 146 per arm), the study has 80% power to demonstrate non-inferiority, defined as a Test group AKI incidence increase of no more than 50% of the Control group rate, assuming a Control incidence of 30%.
DISCUSSION
RBCs are transfused to prevent tissue hypoxia caused by surgery-induced bleeding and anemia. AKI is a sensitive indicator of renal hypoxia and a novel endpoint for assessing RBC efficacy. The ReCePI study is intended to demonstrate the non-inferiority of pathogen-reduced RBCs to conventional RBCs in the support of renal tissue oxygenation due to acute anemia and to characterize the incidence of treatment-related antibodies to RBCs.
Topics: Humans; Prospective Studies; Erythrocytes; Anemia; Cardiac Surgical Procedures; Acute Kidney Injury; Glutathione; Hypoxia; Randomized Controlled Trials as Topic; Multicenter Studies as Topic; Clinical Trials, Phase III as Topic
PubMed: 38082326
DOI: 10.1186/s13063-023-07831-x -
European Journal of Medicinal Chemistry Feb 2024Tacrine (THA), a long withdrawn drug, is still a popular scaffold used in medicinal chemistry, mainly for its good reactivity and multi-targeted effect. However,...
Tacrine (THA), a long withdrawn drug, is still a popular scaffold used in medicinal chemistry, mainly for its good reactivity and multi-targeted effect. However, THA-associated hepatotoxicity is still an issue and must be considered in drug discovery based on the THA scaffold. Following our previously identified hit compound 7-phenoxytacrine (7-PhO-THA), we systematically explored the chemical space with 30 novel derivatives, with a focus on low hepatotoxicity, anticholinesterase action, and antagonism at the GluN1/GluN2B subtype of the NMDA receptor. Applying the down-selection process based on in vitro and in vivo pharmacokinetic data, two candidates, I-52 and II-52, selective GluN1/GluN2B inhibitors thanks to the interaction with the ifenprodil-binding site, have entered in vivo pharmacodynamic studies. Finally, compound I-52, showing only minor affinity to AChE, was identified as a lead candidate with favorable behavioral and neuroprotective effects using open-field and prepulse inhibition tests, along with scopolamine-based behavioral and NMDA-induced hippocampal lesion models. Our data show that compound I-52 exhibits low toxicity often associated with NMDA receptor ligands, and low hepatotoxicity, often related to THA-based compounds.
Topics: Humans; Neuroprotective Agents; Receptors, N-Methyl-D-Aspartate; Tacrine; Cholinesterase Inhibitors; Binding Sites; Cholinesterases; Chemical and Drug Induced Liver Injury; Acetylcholinesterase; Alzheimer Disease; Piperidines
PubMed: 38218127
DOI: 10.1016/j.ejmech.2024.116130