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20 pages, 4288 KiB  
Article
Eisenia bicyclis-Mediated Gold Nanoparticles Exhibit Antibiofilm and Antivirulence Activities Against Pseudomonas aeruginosa and Staphylococcus aureus
by Do Kyung Oh, Du-Min Jo, Nam-Gyun Kim, Kyung-Jin Cho, Geum-Jae Jeong, Nazia Tabassum, Won-Kyo Jung, Fazlurrahman Khan and Young-Mog Kim
Antibiotics 2025, 14(2), 182; https://doi.org/10.3390/antibiotics14020182 - 11 Feb 2025
Cited by 1 | Viewed by 1306
Abstract
Background/Objectives: Brown algae, particularly Eisenia bicyclis, produce various bioactive chemicals with significant application potential in the food, cosmetics, and pharmaceutical industries. This study aimed to evaluate the antibacterial, antibiofilm, and antivirulence properties of the ethyl acetate fraction (EA) of E. bicyclis and [...] Read more.
Background/Objectives: Brown algae, particularly Eisenia bicyclis, produce various bioactive chemicals with significant application potential in the food, cosmetics, and pharmaceutical industries. This study aimed to evaluate the antibacterial, antibiofilm, and antivirulence properties of the ethyl acetate fraction (EA) of E. bicyclis and its synthesized gold nanoparticles (EA-AuNPs), with a focus on their potential applications against both Gram-positive and Gram-negative bacteria. Methods: The bioactive component in the ethyl acetate fraction was identified using a gas chromatography-mass spectroscopy (GC-MS) device and a liquid chromatography-mass spectrometer/mass spectrometry (LC-MS) system. The crystal violet method was utilized to evaluate the biofilm inhibition experiments. Several instruments, including dynamic light scattering, Fourier transform infrared, X-ray diffraction, field emission transmission electron microscopy, and energy-dispersive spectroscopy, were employed to completely characterize the produced EA-AuNPs. The cytotoxicity of the EA-AuNPs was determined using the MTT assay, and the expression of genes linked with biofilm and virulence in Pseudomonas aeruginosa and Staphylococcus aureus was investigated using real-time polymerase chain reaction (RT-PCR). Results: Various bioactive compounds were identified from the EA using GC-MS and LC-MS, including fatty acids and phlorotannins such as eckol, dieckol, 6,6’-bieckol, and phlorofucofuroeckol in high amounts, highlighting EA as a phlorotannin-rich fraction. The EA also demonstrated significant antibiofilm activity, with 79.86% inhibition at 512 μg/mL against P. aeruginosa and 87.00% at 64 μg/mL against S. aureus. EA was then used in the synthesis of gold nanoparticles (AuNPs) to improve their stability and safety. The synthesized EA-AuNPs were determined to have an average size of 165.04 nm, with a zeta potential of −29.86 mV, indicating good stability. In antibiofilm activity assays, EA-AuNPs demonstrated 45.76% inhibition against P. aeruginosa at 1024 μg/mL and 44.64% inhibition against S. aureus at 128 μg/mL. At sub-MIC levels, EA-AuNPs significantly inhibited biofilm formation and virulence factors, including the motility of P. aeruginosa and staphyloxanthin synthesis in S. aureus. The RT-PCR analysis revealed the downregulation of key genes involved in biofilm formation and virulence in P. aeruginosa and S. aureus. Conclusions: These findings highlight the potential of E. bicyclis solvent-soluble extracts and EA-AuNPs as effective antibacterial, antibiofilm, and antivirulence agents, with significant application potential in the pharmaceutical and food industries. To the best of our knowledge, this is the first report of antibiofilm activity against both Gram-positive and Gram-negative bacteria using EA-AuNPs. Full article
(This article belongs to the Special Issue Innovations in Plant-Based Antibiotic and Antiviral Agents)
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27 pages, 5607 KiB  
Article
Experimental Investigation into the Design, Optimization, Toxicity, and Anti-Viral Efficacy of Proliposomes Loaded with Ivermectin Against Infectious Bronchitis Virus Using an Embryonated Chicken Egg Model
by Mohammad H. Alyami, Hamad S. Alyami, Asmaa M. Abdo, Shereen A. Sabry, Shimaa M. G. Mansour, Hanan M. El-Nahas and Margrit M. Ayoub
Pharmaceutics 2025, 17(2), 165; https://doi.org/10.3390/pharmaceutics17020165 - 25 Jan 2025
Viewed by 1350
Abstract
Background: Infectious bronchitis virus (IBV) causes a significant illness in birds, making it a leading source of financial loss in the poultry business. The objective of this study was to assess the effectiveness of proliposomes (PLs) containing ivermectin (IVM) against IBV using [...] Read more.
Background: Infectious bronchitis virus (IBV) causes a significant illness in birds, making it a leading source of financial loss in the poultry business. The objective of this study was to assess the effectiveness of proliposomes (PLs) containing ivermectin (IVM) against IBV using embryonated chicken eggs (ECEs). Methods: A three-factor, two-level (23) full factorial design was employed; carrier/lipid phase ratio (A), stearyl glycyrrhetinate amount (B), and phospholipid type (C) were studied as independent variables, while product yield (PY), entrapment efficiency (EE), particle size (PS), polydispersity index (PDI), zeta potential (ZP), and cumulative percentage of drug released after 6 h (Q6h) were characterized. The selected formulations (PL2 and PL6) were subjected to further characterizations, including IVM toxicity and anti-viral activity. Results: The PY% ranged from 88.6 ± 2.19% to 98.8 ± 0.45%, EE% was from 71.8 ± 2.01% to 96.1 ± 0.51%, PS was from 330.1 ± 55.65 nm to 1801.6 ± 45.61 nm, PDI was from 0.205 ± 0.06 to 0.603 ± 0.03, ZP was from −18.2 ± 0.60 mV to −50.1 ± 1.80 mV, and Q6h was from 80.95 ± 1.36% to 88.79 ± 2.03%. IVM-loaded PLs had lower toxicity in ECEs than pure IVM; the mortality rate was substantially reduced in IBV-infected ECEs injected with PL2 rather than pure IVM. As further evidence of IVM’s anti-viral action against IBV, quantitative real-time polymerase chain reaction (qRT-PCR) revealed that the PL2-treated group exhibited further reduction in IBV’s copies in comparison with the pure IVM-treated group. Conclusions: PLs loaded with IVM are an innovative and potentially effective way to inhibit IBV. Full article
(This article belongs to the Section Drug Delivery and Controlled Release)
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18 pages, 3055 KiB  
Review
POLD3 as Controller of Replicative DNA Repair
by Nabilah Alli, Anna Lou-Hing, Edward L. Bolt and Liu He
Int. J. Mol. Sci. 2024, 25(22), 12417; https://doi.org/10.3390/ijms252212417 - 19 Nov 2024
Cited by 3 | Viewed by 1623
Abstract
Multiple modes of DNA repair need DNA synthesis by DNA polymerase enzymes. The eukaryotic B-family DNA polymerase complexes delta (Polδ) and zeta (Polζ) help to repair DNA strand breaks when primed by homologous recombination or single-strand DNA annealing. DNA synthesis by Polδ and [...] Read more.
Multiple modes of DNA repair need DNA synthesis by DNA polymerase enzymes. The eukaryotic B-family DNA polymerase complexes delta (Polδ) and zeta (Polζ) help to repair DNA strand breaks when primed by homologous recombination or single-strand DNA annealing. DNA synthesis by Polδ and Polζ is mutagenic, but is needed for the survival of cells in the presence of DNA strand breaks. The POLD3 subunit of Polδ and Polζ is at the heart of DNA repair by recombination, by modulating polymerase functions and interacting with other DNA repair proteins. We provide the background to POLD3 discovery, investigate its structure, as well as function in cells. We highlight unexplored structural aspects of POLD3 and new biochemical data that will help to understand the pivotal role of POLD3 in DNA repair and mutagenesis in eukaryotes, and its impact on human health. Full article
(This article belongs to the Special Issue Nuclear Genome Stability: DNA Replication and DNA Repair)
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31 pages, 3629 KiB  
Article
Biocompatible Poly(ε-Caprolactone) Nanocapsules Enhance the Bioavailability, Antibacterial, and Immunomodulatory Activities of Curcumin
by Floriana D’Angeli, Giuseppe Granata, Ivana Roberta Romano, Alfio Distefano, Debora Lo Furno, Antonella Spila, Mariantonietta Leo, Chiara Miele, Dania Ramadan, Patrizia Ferroni, Giovanni Li Volti, Paolo Accardo, Corrada Geraci, Fiorella Guadagni and Carlo Genovese
Int. J. Mol. Sci. 2024, 25(19), 10692; https://doi.org/10.3390/ijms251910692 - 4 Oct 2024
Cited by 3 | Viewed by 1979
Abstract
Curcumin (Cur), the primary curcuminoid found in Curcuma longa L., has garnered significant attention for its potential anti-inflammatory and antibacterial properties. However, its hydrophobic nature significantly limits its bioavailability. Additionally, adipose-derived stem cells (ADSCs) possess immunomodulatory properties, making them useful for treating inflammatory [...] Read more.
Curcumin (Cur), the primary curcuminoid found in Curcuma longa L., has garnered significant attention for its potential anti-inflammatory and antibacterial properties. However, its hydrophobic nature significantly limits its bioavailability. Additionally, adipose-derived stem cells (ADSCs) possess immunomodulatory properties, making them useful for treating inflammatory and autoimmune conditions. This study aims to verify the efficacy of poly(ε-caprolactone) nanocapsules (NCs) in improving Cur’s bioavailability, antibacterial, and immunomodulatory activities. The Cur-loaded nanocapsules (Cur-NCs) were characterized for their physicochemical properties (particle size, polydispersity index, Zeta potential, and encapsulation efficiency) and stability over time. A digestion test simulated the behavior of Cur-NCs in the gastrointestinal tract. Micellar phase analyses evaluated the Cur-NCs’ bioaccessibility. The antibacterial activity of free Cur, NCs, and Cur-NCs against various Gram-positive and Gram-negative strains was determined using the microdilution method. ADSC viability, treated with Cur-NCs and Cur-NCs in the presence or absence of lipopolysaccharide, was analyzed using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide assay. Additionally, ADSC survival was assessed through the Muse apoptotic assay. The expression of both pro-inflammatory (interleukin-1β and tumor necrosis factor-α) and anti-inflammatory (IL-10 and transforming growth factor-β) cytokines on ADSCs was evaluated by real-time polymerase chain reaction. The results demonstrated high stability post-gastric digestion of Cur-NCs and elevated bioaccessibility of Cur post-intestinal digestion. Moreover, Cur-NCs exhibited antibacterial activity against Escherichia coli without affecting Lactobacillus growth. No significant changes in the viability and survival of ADSCs were observed under the experimental conditions. Finally, Cur-NCs modulated the expression of both pro- and anti-inflammatory cytokines in ADSCs exposed to inflammatory stimuli. Collectively, these findings highlight the potential of Cur-NCs to enhance Cur’s bioavailability and therapeutic efficacy, particularly in cell-based treatments for inflammatory diseases and intestinal dysbiosis. Full article
(This article belongs to the Special Issue New Perspective on Inflammatory Diseases: Role of Natural Compounds)
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26 pages, 5993 KiB  
Article
Unveiling the Impact of Eco-Friendly Synthesized Nanoparticles on Vegetative Growth and Gene Expression in Pelargonium graveolens and Sinapis alba L.
by Maha M. Kamel, Abdelfattah Badr, Dalal Hussien M. Alkhalifah, Rehab Mahmoud, Yasser GadelHak and Wael N. Hozzein
Molecules 2024, 29(14), 3394; https://doi.org/10.3390/molecules29143394 - 19 Jul 2024
Viewed by 1880
Abstract
Nanoscale geranium waste (GW) and magnesium nanoparticle/GW nanocomposites (Mg NP/GW) were prepared using green synthesis. The Mg NP/GW samples were subjected to characterization using X-ray diffraction (XRD) and Fourier-transform infrared spectroscopy (FTIR-FT). The surface morphology of the materials was examined using a scanning [...] Read more.
Nanoscale geranium waste (GW) and magnesium nanoparticle/GW nanocomposites (Mg NP/GW) were prepared using green synthesis. The Mg NP/GW samples were subjected to characterization using X-ray diffraction (XRD) and Fourier-transform infrared spectroscopy (FTIR-FT). The surface morphology of the materials was examined using a scanning electron microscope (SEM), and their thermal stability was assessed through thermal gravimetric analysis (TG). The BET-specific surface area, pore volume, and pore size distribution of the prepared materials were determined using the N2 adsorption–desorption method. Additionally, the particle size and zeta potentials of the materials were also measured. The influence of the prepared nanomaterials on seed germination was intensively investigated. The results revealed an increase in seed germination percent at low concentrations of Mg NP/GWs. Upon treatment with Mg NP/GW nanoparticles, a reduction in the mitotic index (MI) was observed, indicating a decrease in cell division. Additionally, an increase in chromosomal abnormalities was detected. The efficacy of GW and Mg NP/GW nanoparticles as new elicitors was evaluated by studying their impact on the expression levels of the farnesyl diphosphate synthase (FPPS1) and geranylgeranyl pyrophosphate (GPPS1) genes. These genes play a crucial role in the terpenoid biosynthesis pathway in Sinapis alba (S. alba) and Pelargonium graveolens (P. graveolens) plants. The expression levels were analyzed using reverse transcription–quantitative polymerase chain reaction (RT-qPCR) analysis. The qRT-PCR analysis of FPPS and GPPS gene expression was performed. The outputs of FPPS1 gene expression demonstrated high levels of mRNA in both S. alba and P. graveolens with fold changes of 25.24 and 21.68, respectively. In contrast, the minimum expression levels were observed for the GPPS1 gene, with fold changes of 11.28 and 6.48 in S. alba and P. graveolens, respectively. Thus, this study offers the employment of medicinal plants as an alternative to fertilizer usage resulting in promoting environmental preservation, optimal waste utilization, reducing water consumption, and cost reduction. Full article
(This article belongs to the Topic Advanced Biomaterials: Processing and Applications)
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19 pages, 8076 KiB  
Article
Antibacterial Effect of Silver Nanoparticles against Oral Biofilms in Subjects with Motor and Intellectual Disabilities
by Carolina Holguín-Meráz, Rita Elizabeth Martínez-Martínez, Erasto Armando Zaragoza-Contreras, Rubén Abraham Domínguez-Pérez, Simón Yobanny Reyes-López, Alejandro Donohue-Cornejo, Juan Carlos Cuevas-González, Erika de Lourdes Silva-Benítez, Nelly Molina-Frechero and León Francisco Espinosa-Cristóbal
J. Funct. Biomater. 2024, 15(7), 191; https://doi.org/10.3390/jfb15070191 - 10 Jul 2024
Cited by 3 | Viewed by 1629
Abstract
Background: Motor and intellectual disabilities (MIDs) represent a great challenge for maintaining general health due to physical and cognitive limitations, particularly in the maintenance and preservation of oral health. Silver nanoparticles (AgNPs) have emerged as a promising therapeutic tool for bacterial control, including [...] Read more.
Background: Motor and intellectual disabilities (MIDs) represent a great challenge for maintaining general health due to physical and cognitive limitations, particularly in the maintenance and preservation of oral health. Silver nanoparticles (AgNPs) have emerged as a promising therapeutic tool for bacterial control, including oral biofilms; however, knowledge of the bactericidal effectiveness of oral biofilms from patients with MIDs is insufficient. This study aims to determine the antimicrobial effect of AgNPs on different oral biofilms taken from patients with and without MIDs. Methods: Two sizes of AgNPs were prepared and characterized by dynamic light scattering (DLS) and transmission electron microscopy (TEM). Through consecutive sampling, biofilm samples were collected from 17 subjects with MIDs and 20 subjects without disorders. The antimicrobial effect was determined by obtaining the minimum inhibitory concentration (MIC) of AgNPs, and the identification and distribution of oral bacterial species were determined by polymerase chain reaction (PCR). Finally, correlations between sociodemographic characteristics and the antimicrobial levels of AgNPs were also explored. The values of the MIC results were analyzed with IBM-SPSS software (version25) using non-parametric tests for independent groups and correlations, with statistical significance being considered as p < 0.05. Results: Both sizes of AgNPs exhibited tight particle size distributions (smaller: 10.2 ± 0.7 nm; larger: 29.3 ± 2.3 nm) with zeta potential values (−35.0 ± 3.3 and −52.6 ± 8.5 mV, respectively) confirming the stability that resulted in little to no agglomeration of nanoparticles. Although both sizes of AgNPs had good antimicrobial activity in all oral biofilms, the smallest particles had the best antimicrobial effects on the oral biofilm samples from patients with and without MIDs, even better than chlorhexidine (CHX) (p < 0.05). Likewise, the patients with disabilities showed higher levels of antimicrobial sensitivity to AgNPs compared with CHX (p < 0.05). Although the microorganisms included in the biofilms of females had a statistically higher growth level, the AgNP antimicrobial effect was statistically similar in both genders (p > 0.05). The most frequent bacteria for all oral biofilms were S. mutans (100%), P. intermedia (91.6%), T. forsythia (75.0%), T. denticola (75.0%), P. gingivalis (66.6%), F. nucleatum (66.6%), S. sobrinus (50.0%), and A. actinomycetemcomitans (8.3%). Conclusions: AgNPs exhibited considerable antimicrobial potential to be used as a complementary and alternative tool in maintaining and preserving oral health in patients with MIDs. Full article
(This article belongs to the Special Issue Antibacterial Biomaterials for Medical Applications)
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21 pages, 6374 KiB  
Article
Microsponge-Based Gel Loaded with Immunosuppressant as a Simple and Valuable Strategy for Psoriasis Therapy: Determination of Pro-Inflammatory Response through Cytokine IL-2 mRNA Expression
by Yasir Mehmood, Hira Shahid, Umar Inzamam ul Huq, Hamza Rafeeq, Hafiz Muhammad Bilal Khalid, Mohammad N. Uddin and Mohsin Kazi
Gels 2023, 9(11), 871; https://doi.org/10.3390/gels9110871 - 1 Nov 2023
Cited by 5 | Viewed by 3061
Abstract
Tacrolimus (TL) is a topical calcineurin inhibitor immunosuppressive drug widely used to manage various skin disorders. Herein, we report a TL-loaded microsphere gel formulation with severe atopic dermatitis effects that are required to manage skin disorders. The current study adopted a modified emulsion [...] Read more.
Tacrolimus (TL) is a topical calcineurin inhibitor immunosuppressive drug widely used to manage various skin disorders. Herein, we report a TL-loaded microsphere gel formulation with severe atopic dermatitis effects that are required to manage skin disorders. The current study adopted a modified emulsion solvent evaporation technique to synthesize TL-loaded microspheres, which were further converted into gels for skin use. Characterization of the synthesized formulation was performed by differential dynamic light scattering, scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy, X-ray crystallography, Brunauer–Emmett–Teller (BET) analysis, differential scanning calorimetry, and drug release. A Franz diffusion cell was used to study the diffusion of TL for up to 8 h at pH 6.8 and 5.5. Evaluation of cell viability was determined by MTT assay and showed higher IC50 values compared to the plain drug. RNA extraction, real-time polymerase chain reaction (RT–PCR), and reverse transcription were also performed to determine the expression levels of the anti-inflammatory cytokine IL-2. Particle size determination was performed by a zeta sizer, and the TL microsphere size was 1745 ± 70 nm with a good polydispersity (0.337 ± 0.12). The drug entrapment efficiency was also very good at 60% ± 10, and the drug release was 93.9% ± 3.5 within 8 h. An in vitro diffusion study of the formulation also showed improved permeability at both pH values (4.5 and 5.5). The findings of the hemolytic tests demonstrated that TL-MG at concentrations of 50, 100, and 200 mg/mL did not produce any hemolysis. A dose-dependent pattern of cytotoxicity was found during the cell viability assay, with an IC50 value of 787.55 ± 12.78 µg/mL. There was a significant decrease in the IL-2 level in the TL-MG group compared to the other groups. TL-MG microspheres were nontoxic carriers for tacrolimus delivery, with greater loading capacity, a significant release profile, and enhanced cellular uptake with improved permeability. Full article
(This article belongs to the Special Issue Advances in Hydrogels for Biomedical Applications)
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28 pages, 5349 KiB  
Article
Liposome Formulations for the Strategic Delivery of PARP1 Inhibitors: Development and Optimization
by Carlota J. F. Conceição, Elin Moe, Paulo A. Ribeiro and Maria Raposo
Nanomaterials 2023, 13(10), 1613; https://doi.org/10.3390/nano13101613 - 11 May 2023
Cited by 5 | Viewed by 2938
Abstract
The development of a lipid nano-delivery system was attempted for three specific poly (ADP-ribose) polymerase 1 (PARP1) inhibitors: Veliparib, Rucaparib, and Niraparib. Simple lipid and dual lipid formulations with 1,2-dipalmitoyl-sn-glycero-3-phospho-rac-(1′-glycerol) sodium salt (DPPG) and 1,2-dipalmitoyl-sn-glycero-3-phosphocoline (DPPC) were developed and tested following the thin-film [...] Read more.
The development of a lipid nano-delivery system was attempted for three specific poly (ADP-ribose) polymerase 1 (PARP1) inhibitors: Veliparib, Rucaparib, and Niraparib. Simple lipid and dual lipid formulations with 1,2-dipalmitoyl-sn-glycero-3-phospho-rac-(1′-glycerol) sodium salt (DPPG) and 1,2-dipalmitoyl-sn-glycero-3-phosphocoline (DPPC) were developed and tested following the thin-film method. DPPG-encapsulating inhibitors presented the best fit in terms of encapsulation efficiency (>40%, translates into concentrations as high as 100 µM), zeta potential values (below −30 mV), and population distribution (single population profile). The particle size of the main population of interest was ~130 nm in diameter. Kinetic release studies showed that DPPG-encapsulating PARP1 inhibitors present slower drug release rates than liposome control samples, and complex drug release mechanisms were identified. DPPG + Veliparib/Niraparib presented a combination of diffusion-controlled and non-Fickian diffusion, while anomalous and super case II transport was verified for DPPG + Rucaparib. Spectroscopic analysis revealed that PARP1 inhibitors interact with the DPPG lipid membrane, promoting membrane water displacement from hydration centers. A preferential membrane interaction with lipid carbonyl groups was observed through hydrogen bonding, where the inhibitors’ protonated amine groups may be the major players in the PARP1 inhibitor encapsulation mode. Full article
(This article belongs to the Special Issue Application of Lipid Nanoparticles in Drug and Gene Delivery)
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16 pages, 3081 KiB  
Article
Antifungal Properties of Biogenic Selenium Nanoparticles Functionalized with Nystatin for the Inhibition of Candida albicans Biofilm Formation
by Shivraj Hariram Nile, Dipalee Thombre, Amruta Shelar, Krithika Gosavi, Jaiprakash Sangshetti, Weiping Zhang, Elwira Sieniawska, Rajendra Patil and Guoyin Kai
Molecules 2023, 28(4), 1836; https://doi.org/10.3390/molecules28041836 - 15 Feb 2023
Cited by 25 | Viewed by 3846
Abstract
In the present study, biogenic selenium nanoparticles (SeNPs) have been prepared using Paenibacillus terreus and functionalized with nystatin (SeNP@PVP_Nystatin nanoconjugates) for inhibiting growth, morphogenesis, and a biofilm in Candida albicans. Ultraviolet–visible spectroscopy analysis has shown a characteristic absorption at 289, 303, and [...] Read more.
In the present study, biogenic selenium nanoparticles (SeNPs) have been prepared using Paenibacillus terreus and functionalized with nystatin (SeNP@PVP_Nystatin nanoconjugates) for inhibiting growth, morphogenesis, and a biofilm in Candida albicans. Ultraviolet–visible spectroscopy analysis has shown a characteristic absorption at 289, 303, and 318 nm, and X-ray diffraction analysis has shown characteristic peaks at different 2θ values for SeNPs. Electron microscopy analysis has shown that biogenic SeNPs are spherical in shape with a size in the range of 220–240 nm. Fourier transform infrared spectroscopy has confirmed the functionalization of nystatin on SeNPs (formation of SeNP@PVP_Nystatin nanoconjugates), and the zeta potential has confirmed the negative charge on the nanoconjugates. Biogenic SeNPs are inactive; however, nanoconjugates have shown antifungal activities on C. albicans (inhibited growth, morphogenesis, and a biofilm). The molecular mechanism for the action of nanoconjugates via a real-time polymerase chain reaction has shown that genes involved in the RAS/cAMP/PKA signaling pathway play an important role in antifungal activity. In cytotoxic studies, nanoconjugates have inhibited only 12% growth of the human embryonic kidney cell line 293 cells, indicating that the nanocomposites are not cytotoxic. Thus, the biogenic SeNPs produced by P. terreus can be used as innovative and effective drug carriers to increase the antifungal activity of nystatin. Full article
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14 pages, 1888 KiB  
Article
A Role for the Interactions between Polδ and PCNA Revealed by Analysis of pol3-01 Yeast Mutants
by Shaked Nir Heyman, Mika Golan, Batia Liefshitz and Martin Kupiec
Genes 2023, 14(2), 391; https://doi.org/10.3390/genes14020391 - 2 Feb 2023
Viewed by 2663
Abstract
Several DNA polymerases participate in DNA synthesis during genome replication and DNA repair. PCNA, a homotrimeric ring, acts as a processivity factor for DNA polymerases. PCNA also acts as a “landing pad” for proteins that interact with chromatin and DNA at the moving [...] Read more.
Several DNA polymerases participate in DNA synthesis during genome replication and DNA repair. PCNA, a homotrimeric ring, acts as a processivity factor for DNA polymerases. PCNA also acts as a “landing pad” for proteins that interact with chromatin and DNA at the moving fork. The interaction between PCNA and polymerase delta (Polδ) is mediated by PIPs (PCNA-interacting peptides), in particular the one on Pol32, a regulatory subunit of Polδ. Here, we demonstrate that pol3-01, an exonuclease mutant of Polδ’s catalytic subunit, exhibits a weak interaction with Pol30 compared to the WT DNA polymerase. The weak interaction activates DNA bypass pathways, leading to increased mutagenesis and sister chromatid recombination. Strengthening pol3-01′s weak interaction with PCNA suppresses most of the phenotypes. Our results are consistent with a model in which Pol3-01 tends to detach from the chromatin, allowing an easier replacement of Polδ by the trans-lesion synthesis polymerase Zeta (Polz), thus leading to the increased mutagenic phenotype. Full article
(This article belongs to the Special Issue Mechanisms of DNA Damage, Repair and Mutagenesis 2023)
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25 pages, 3823 KiB  
Article
Nanocrystallization Improves the Solubilization and Cytotoxic Effect of a Poly (ADP-Ribose)-Polymerase-I Inhibitor
by Amer S. Alali, Mohd Abul Kalam, Mohammed Muqtader Ahmed, M. Ali Aboudzadeh, Sulaiman S. Alhudaithi, Md. Khalid Anwer, Farhat Fatima and Muzaffar Iqbal
Polymers 2022, 14(22), 4827; https://doi.org/10.3390/polym14224827 - 9 Nov 2022
Cited by 12 | Viewed by 3047
Abstract
Olaparib (OLA) is an anticancer agent that acts by inhibiting the poly (ADP-ribose)-polymerase-I (PARP-I). Due to its low solubility and low permeability, it has been placed as a BCS Class-IV drug and hence its clinical use is limited. In this study, we develop [...] Read more.
Olaparib (OLA) is an anticancer agent that acts by inhibiting the poly (ADP-ribose)-polymerase-I (PARP-I). Due to its low solubility and low permeability, it has been placed as a BCS Class-IV drug and hence its clinical use is limited. In this study, we develop the nanocrystals of OLA as a way to improve its solubility and other performances. The OLA-NCs were prepared by antisolvent precipitation method through homogenization and probe sonication technique using a novel amphiphilic polymeric stabilizer (Soluplus®). Particle characterization resulted approximately 103.13 nm, polydispersity-index was 0.104 with positive zeta-potential of +8.67 mV. The crystal morphology by SEM of OLA-NCs (with and without mannitol) exhibited nano-crystalline prism-like structures as compared to the elongated OLA-pure. The DSC, XRD and FTIR were performed to check the interaction of Soluplus, mannitol and OLA did not exhibit any physical interaction among the OLA, Soluplus® and mannitol that is indicated by the presence of parent wave number peak. Two-fold increased solubility of OLA was found in PBS with Soluplus® from the NCs (69.3 ± 6.2 µgmL−1) as compared to pure drug (35.6 ± 7.2 µgmL−1). In vitro release of drug from OLA-NCs was higher (78.2%) at 12 h at pH 6.8 and relatively lower (53.1%) at pH 1.2. In vitro cellular cytotoxicity and anticancer effects were examined on MCF-7 cells. OLA-NCs were found effectively potent to MCF-7 cells compared with OLA-pure with approximately less than half IC50 value during MTT assay. Estimation of p53, Caspase-3 and Caspase-9 in MCF-7 cells indicated that OLA-NCs have significantly (p < 0.05) increased their expressions. After single oral dose in rats, 12 h plasma drug concentration-time profile indicated approximately 2.06-, 2.29-, 2–25- and 2.62-folds increased Cmax, AUC0-12 h, AUC0-∞ and AUMC0-∞, respectively, from the NCs as compared to OLA-pure. Storage stability indicated that the OLA-NCs was physically and chemically stable at 4 °C, 25 °C and 40 °C up to 6-months. Overall, OLA-NCs were deliberated; its potential feasibility to overwhelm the formulation challenges related to poorly soluble drugs and its future clinical applications. Full article
(This article belongs to the Special Issue Function of Polymers in Encapsulation Process II)
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17 pages, 11440 KiB  
Article
Zinc Oxide Nanoparticles as Potential Delivery Carrier: Green Synthesis by Aspergillus niger Endophytic Fungus, Characterization, and In Vitro/In Vivo Antibacterial Activity
by Dalia H. Abdelkader, Walaa A. Negm, Engy Elekhnawy, Duaa Eliwa, Basmah N. Aldosari and Alanood S. Almurshedi
Pharmaceuticals 2022, 15(9), 1057; https://doi.org/10.3390/ph15091057 - 26 Aug 2022
Cited by 58 | Viewed by 4552
Abstract
We aimed to synthesize zinc oxide nanoparticles (ZnO NPs) using the endophytic fungal extract of Aspergillus niger. The prepared ZnO NPs were characterized, and their in vitro and in vivo antibacterial activity was investigated. Isolated endophytic fungus identification was carried out using [...] Read more.
We aimed to synthesize zinc oxide nanoparticles (ZnO NPs) using the endophytic fungal extract of Aspergillus niger. The prepared ZnO NPs were characterized, and their in vitro and in vivo antibacterial activity was investigated. Isolated endophytic fungus identification was carried out using 18S rRNA. A. niger endophytic fungal extract was employed for the green synthesis of ZnO NPs. The in vitro antibacterial activity of the prepared ZnO NPs was elucidated against Staphylococcus aureus using the broth microdilution method and quantitative real-time polymerase chain reaction (qRT-PCR). Additionally, the in vivo antibacterial activity was elucidated using a systemic infection model in mice. The biosynthesized ZnO NPs showed a maximum optical density at 380 nm with characteristic peaks on the Fourier-transform infrared spectrum. The X-ray diffraction pattern was highly matched with a standard platform of zinc oxide crystals. Energy-dispersive X-ray analysis confirmed that the main composition of nanoparticles was zinc and oxygen atoms. Scanning and transmission electron microscopies showed spherical geometry with a smooth surface. Zeta potential measurements (26.6 ± 0.56 mV) verified the adequate stability of ZnO NPs. Minimum inhibitory concentrations of ZnO NPs against S. aureus isolates ranged from 8 to 128 µg/mL. Additionally, ZnO NPs revealed antibiofilm activity, resulting in the downregulation of the tested biofilm genes in 29.17% of S. aureus isolates. Regarding the in vivo experiment, ZnO NPs reduced congestion and fibrosis in liver and spleen tissues. They also improved liver function, increased the survival rate, and significantly decreased inflammatory markers (p < 0.05). ZnO NPs synthesized by A. niger endophytic fungus revealed a promising in vivo and in vitro antibacterial action against S. aureus isolates. Full article
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18 pages, 2130 KiB  
Article
Modulation of Chitosan-TPP Nanoparticle Properties for Plasmid DNA Vaccines Delivery
by Renato Nunes, Ana Sofia Serra, Aiva Simaite and Ângela Sousa
Polymers 2022, 14(7), 1443; https://doi.org/10.3390/polym14071443 - 1 Apr 2022
Cited by 22 | Viewed by 5124
Abstract
Nucleic acid vaccines have become a revolutionary technology to give a fast, safe, cost-effective and efficient response against viral infections, such as SARS-CoV-2 or Human papillomavirus (HPV). However, to ensure their effectiveness, the development of adequate methods to protect, carry, and deliver nucleic [...] Read more.
Nucleic acid vaccines have become a revolutionary technology to give a fast, safe, cost-effective and efficient response against viral infections, such as SARS-CoV-2 or Human papillomavirus (HPV). However, to ensure their effectiveness, the development of adequate methods to protect, carry, and deliver nucleic acids is fundamental. In this work, nanoparticles (NPs) of chitosan (CS)-tripolyphosphate (TPP)-plasmid DNA (pDNA) were thoroughly modulated and characterized, by measuring the charge and size through dynamic light scattering (DLS) and morphology by scanning electron microscopy (SEM). Stability, cytotoxicity and cellular uptake of NPs were also evaluated. Finally, the effect of polyplexes on the expression of HPV E7 antigen in human fibroblast and RAW cells was investigated through polymerase chain reaction (PCR) and real-time PCR. The results showed NPs with a spherical/oval shape, narrow size distribution <180 nm and positive zeta potentials (>20 mV) and good stability after one month of storage at 4 °C in formulation buffer or when incubated in culture medium and trypsin. In vitro studies of NPs cytotoxicity revealed that the elimination of formulation buffers led to an improvement in the rate of cell viability. The E7 antigen transcription was also increased for NPs obtained with high pDNA concentration (60 μg/mL). The analyzed CS-TPP-pDNA polyplexes can offer a promising vehicle for nucleic acid vaccines, not only in the prevention or treatment of viral infections, but also to fight emergent and future pathogens. Full article
(This article belongs to the Special Issue Biopolymers: Recent Progress and New Perspectives II)
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15 pages, 3009 KiB  
Article
Disrupting the MAD2L2-Rev1 Complex Enhances Cell Death upon DNA Damage
by Nomi Pernicone, Maria Elias, Itay Onn, Dror Tobi and Tamar Listovsky
Molecules 2022, 27(3), 636; https://doi.org/10.3390/molecules27030636 - 19 Jan 2022
Cited by 8 | Viewed by 3261
Abstract
DNA-damaging chemotherapy agents such as cisplatin have been the first line of treatment for cancer for decades. While chemotherapy can be very effective, its long-term success is often reduced by intrinsic and acquired drug resistance, accompanied by chemotherapy-resistant secondary malignancies. Although the mechanisms [...] Read more.
DNA-damaging chemotherapy agents such as cisplatin have been the first line of treatment for cancer for decades. While chemotherapy can be very effective, its long-term success is often reduced by intrinsic and acquired drug resistance, accompanied by chemotherapy-resistant secondary malignancies. Although the mechanisms causing drug resistance are quite distinct, they are directly connected to mutagenic translesion synthesis (TLS). The TLS pathway promotes DNA damage tolerance by supporting both replication opposite to a lesion and inaccurate single-strand gap filling. Interestingly, inhibiting TLS reduces both cisplatin resistance and secondary tumor formation. Therefore, TLS targeting is a promising strategy for improving chemotherapy. MAD2L2 (i.e., Rev7) is a central protein in TLS. It is an essential component of the TLS polymerase zeta (ζ), and it forms a regulatory complex with Rev1 polymerase. Here we present the discovery of two small molecules, c#2 and c#3, that directly bind both in vitro and in vivo to MAD2L2 and influence its activity. Both molecules sensitize lung cancer cell lines to cisplatin, disrupt the formation of the MAD2L2-Rev1 complex and increase DNA damage, hence underlining their potential as lead compounds for developing novel TLS inhibitors for improving chemotherapy treatments. Full article
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18 pages, 2708 KiB  
Article
Size-Dependent Cytotoxic and Molecular Study of the Use of Gold Nanoparticles against Liver Cancer Cells
by Abdulaziz A. Al-Khedhairy and Rizwan Wahab
Appl. Sci. 2022, 12(2), 901; https://doi.org/10.3390/app12020901 - 17 Jan 2022
Cited by 13 | Viewed by 3250
Abstract
The size of nanomaterials influences physicochemical parameters, and variations in the size of nanomaterials can have a significant effect on their biological activities in cells. Due to the potential applicability of nanoparticles (NPs), the current work was designed to carry out a size-dependent [...] Read more.
The size of nanomaterials influences physicochemical parameters, and variations in the size of nanomaterials can have a significant effect on their biological activities in cells. Due to the potential applicability of nanoparticles (NPs), the current work was designed to carry out a size-dependent study of gold nanoparticles (GNPs) in different dimensions, synthesized via a colloidal solution process. Three dissimilar-sized GNPs, GNPs-1 (10–15 nm), GNPs-2 (20–30 nm), and GNPs-3 (45 nm), were prepared and characterized via transmission electron microscopy (TEM), high-resolution TEM (HR-TEM), hydrodynamic size, zeta potential, and UV-visible spectroscopy, and applied against liver cancer (HepG2) cells. Various concentrations of GNPs (1, 2, 5, 10, 50, and 100 µg/mL) were applied against the HepG2 cancer cells to assess the percentage of cell viability via MTT and NRU assays; reactive oxygen species (ROS) generation was also used. ROS generation was increased by 194%, 164%, and 153% for GNPs-1, GNPs-2, and GNPs-3, respectively, in the HepG2 cells. The quantitative polymerase chain reaction (qPCR) data for the HepG2 cells showed up-regulation in gene expression of apoptotic genes (Bax, p53, and caspase-3) when exposed to the different-sized GNPs, and defined their respective roles. Based on the results, it was concluded that GNPs of different sizes have the potential to induce cancer cell death. Full article
(This article belongs to the Special Issue Applications of Nanotechnology in Cancer)
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