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Keywords = strain isolation pad

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14 pages, 3913 KB  
Article
Isolation of Porcine Adenovirus Serotype 5 and Construction of Recombinant Virus as a Vector Platform for Vaccine Development
by Qianhua He, Jun Wu, Zhilong Bian, Yuan Sun and Jingyun Ma
Viruses 2025, 17(9), 1270; https://doi.org/10.3390/v17091270 - 19 Sep 2025
Viewed by 207
Abstract
Porcine adenovirus serotype 5 (PAdV-5) is an emerging viral vector platform for veterinary vaccines; however, its genomic plasticity and essential replication elements remain incompletely characterized. This study reports the isolation and reverse genetic manipulation of a novel PAdV-5 strain (GD84) from diarrheic piglets [...] Read more.
Porcine adenovirus serotype 5 (PAdV-5) is an emerging viral vector platform for veterinary vaccines; however, its genomic plasticity and essential replication elements remain incompletely characterized. This study reports the isolation and reverse genetic manipulation of a novel PAdV-5 strain (GD84) from diarrheic piglets in China. PCR screening of 167 clinical samples revealed a PAdV-5 detection rate of 38.3% (64/167), with successful isolation on ST cells after three blind passages. The complete GD84 genome is 32,620 bp in length and exhibited 99.0% nucleotide identity to the contemporary strain Ino5, but only 97.0% to the prototype HNF-70. It features an atypical GC content of 51.0% and divergent structural genes—most notably the hexon gene (89% identity to HNF-70)—suggesting altered immunogenicity. Using Red/ET recombineering, we established a rapid (less than 3 weeks) reverse genetics platform and generated four E3-modified recombinants: ΔE3-All-eGFP, ΔE3-12.5K-eGFP, ΔE3-12.5K+ORF4-eGFP, and E3-Insert-eGFP. Crucially, the ΔE3-All-eGFP construct (complete E3 deletion) failed to be rescued, while constructs preserving the 12.5K open reading frame (ORF) yielded replication-competent viruses with sustained eGFP expression over three serial passages and titers over 107.0 TCID50/mL. Fluorescence intensity was inversely correlated with genome size, as the full-length E3-Insert-eGFP virus showed reduced expression compared with the ΔE3 variants. Our work identifies the 12.5K ORF as essential for PAdV-5 replication and provides an optimized vaccine engineering platform that balances genomic payload capacity with replicative fitness. Full article
(This article belongs to the Section Animal Viruses)
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15 pages, 987 KB  
Article
In Vitro Interactions Between Bacteriophages and Antibacterial Agents of Various Classes Against Multidrug-Resistant Metallo-β-Lactamase-Producing Pseudomonas aeruginosa Clinical Isolates
by Paschalis Paranos, Sophia Vourli, Spyros Pournaras and Joseph Meletiadis
Pharmaceuticals 2025, 18(3), 343; https://doi.org/10.3390/ph18030343 - 27 Feb 2025
Cited by 4 | Viewed by 1131
Abstract
Background: Combination therapy with antibiotics and phages has been suggested to increase the antibacterial activity of both antibiotics and phages. We tested the in vitro activity of five antibiotics belonging to different classes in combination with lytic bacteriophages against multidrug-resistant metallo-β-lactamase (MBL)-producing Pseudomonas [...] Read more.
Background: Combination therapy with antibiotics and phages has been suggested to increase the antibacterial activity of both antibiotics and phages. We tested the in vitro activity of five antibiotics belonging to different classes in combination with lytic bacteriophages against multidrug-resistant metallo-β-lactamase (MBL)-producing Pseudomonas aeruginosa isolates. Material/Methods: A total of 10 non-repetitive well-characterized MBL-producing P. aeruginosa isolates (5 NDM, 5 VIM) co-resistant to aminoglycosides and quinolones were used. Phage–antibiotic interactions were assessed using an ISO-20776-based broth microdilution checkerboard assay in 96-well microtitration plates. Two-fold dilutions of colistin (8–0.125 mg/L), ciprofloxacin, meropenem, aztreonam, and amikacin (256–4 mg/L) were combined with ten-fold dilutions of five different phages (5 × 109–5 × 100 PFU/mL) belonging to Pakpunavirus, Phikzvirus, Pbunavirus, and Phikmvvirus genus. Plates were incubated at 35 ± 2 °C for 24 h, and the minimum inhibitory concentration of antibiotics (MICA) and phages (MICP) were determined as the lowest drug and phage concentration, resulting in <10% growth based on photometric reading at 550 nm. Interactions were assessed based on the fractional inhibitory concentration index (FICi) of three independent replicates and clinical relevance based on the reversal of phenotypic resistance. The statistical significance of each drug alone and in combination with phages was assessed using GraphPad Prism 8.0. Results: Synergistic and additive interactions were found for 60–80% of isolates for all drugs. FICis were statistically significantly lower than 0.5 for colistin (p = 0.005), ciprofloxacin (p = 0.02), meropenem (p = 0.003), and amikacin (p = 0.002). Interactions were found at clinically achievable concentrations for colistin, meropenem, and amikacin, and a reversal of phenotypic resistance was observed for most strains (63–64%) for amikacin and meropenem. Antagonism was found for few isolates with all antibiotics tested. Phage vB_PaerM_AttikonH10 and vB_PaerP_AttikonH4 belonging to Phikzvirus and Phikmvvirus genus, respectively, showed either synergistic (FICi ≤ 0.35) or additive effects with most antibiotics tested. Conclusions: Synergy was observed for most drugs and phages with amikacin, showing strong synergy and reversal of phenotypic resistance against most isolates. Taking into account the wide utility of jumbo phages obtained, the findings of vB_PaerM_AttikonH10 in combination with different classes of antibiotics can enhance the activity of currently ineffective antibiotics against MBL-producing P. aeruginosa isolates. Full article
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12 pages, 279 KB  
Article
Six-Year Epidemiologic Analysis of Antibiotic Resistance Patterns of Klebsiella pneumoniae Infections in a Tertiary Healthcare Center in Western Romania
by Elena Hogea, Alexandra-Cristina Muntean, Oana Plavitu, Adelina Fratutu, Iulia Georgiana Bogdan, Felix Bratosin, Cristian-Iulian Oancea, Septimiu-Radu Susa, Daniel Porav-Hodade, Mihai-Calin Bica and Alexandru Blidisel
Microbiol. Res. 2024, 15(4), 2661-2672; https://doi.org/10.3390/microbiolres15040176 (registering DOI) - 12 Dec 2024
Viewed by 1384
Abstract
Background and Objectives: Klebsiella pneumoniae is a major nosocomial pathogen with a remarkable ability to develop resistance to multiple antibiotics, posing significant treatment challenges. This study aims to evaluate the antimicrobial resistance patterns among multidrug-resistant (MDR) and non-MDR strains of K. pneumoniae isolated [...] Read more.
Background and Objectives: Klebsiella pneumoniae is a major nosocomial pathogen with a remarkable ability to develop resistance to multiple antibiotics, posing significant treatment challenges. This study aims to evaluate the antimicrobial resistance patterns among multidrug-resistant (MDR) and non-MDR strains of K. pneumoniae isolated over a six-year period (2018–2023) at the Clinical Hospital of Infectious Diseases and Pulmonology “Dr. Victor Babes” in Timisoara, Romania. The objectives include categorizing isolates based on their antibiotic resistance profiles and identifying trends in resistance to key antibiotics to optimize treatment strategies and enhance infection control measures. Materials and Methods: A cross-sectional analysis was conducted on K. pneumoniae isolates obtained from various clinical samples between January 2018 and December 2023. Identification was performed using standard bacteriological procedures, and antimicrobial susceptibility testing was conducted using the Kirby–Bauer disk diffusion method in accordance with EUCAST guidelines. Isolates were classified as susceptible, resistant, MDR, extensively drug-resistant (XDR), or pandrug-resistant (PDR) based on ECDC definitions. Data were analyzed using GraphPad Prism 6, with chi-square tests and Cochran–Armitage trend tests applied where appropriate. Statistical significance was set at p < 0.05. Results: A total of 1,081 K. pneumoniae isolates were identified over the six-year period, increasing from 118 isolates in 2018 to 319 in 2023. The proportion of XDR and PDR strains showed a significant upward trend from 30.5% in 2018 to 57.4% in 2023 (p < 0.001). Specifically, XDR strains increased from 22.9% in 2018 to 39.8% in 2023, while PDR strains rose from 7.6% to 17.6%. Among monomicrobial infections in 2023, XDR and PDR strains accounted for 42.4% and 16.5%, respectively. Resistance to carbapenems also showed a significant increase; for instance, resistance to ertapenem rose from 35.6% in 2018 to 54.2% in 2023 (p < 0.001). Subgroup analysis revealed that isolates from bronchial aspirates had the highest rates of XDR and PDR strains in 2023, at 38.0% and 17.2%, respectively. Additionally, polymicrobial infections where both K. pneumoniae and co-infecting pathogens were XDR/PDR increased from 24.2% in 2018 to 46.6% in 2023 (p < 0.001). Conclusions: The study demonstrates a significant escalation in antimicrobial resistance among K. pneumoniae isolates over the six-year period, particularly in XDR and PDR strains. The rising trend of resistance to critical antibiotics like carbapenems underscores the urgent need for enhanced antimicrobial stewardship and infection control measures. Targeted interventions are essential to curb the spread of these resistant strains and to optimize therapeutic strategies. Full article
21 pages, 6495 KB  
Article
The Coexistence of Klebsiella pneumoniae and Candida albicans Enhanced Biofilm Thickness but Induced Less Severe Neutrophil Responses and Less Inflammation in Pneumonia Mice Than K. pneumoniae Alone
by Pornpimol Phuengmaung, Chiratchaya Chongrak, Wilasinee Saisorn, Jiradej Makjaroen, Uthaibhorn Singkham-in and Asada Leelahavanichkul
Int. J. Mol. Sci. 2024, 25(22), 12157; https://doi.org/10.3390/ijms252212157 - 12 Nov 2024
Cited by 1 | Viewed by 2366
Abstract
Due to the possible coexistence of Klebsiella pneumoniae (KP) and Candida albicans (CA), strains of KP and CA with biofilm production properties clinically isolated from patients were tested. The production of biofilms from the combined organisms (KP+CA) was higher than the biofilms from [...] Read more.
Due to the possible coexistence of Klebsiella pneumoniae (KP) and Candida albicans (CA), strains of KP and CA with biofilm production properties clinically isolated from patients were tested. The production of biofilms from the combined organisms (KP+CA) was higher than the biofilms from each organism alone, as indicated by crystal violet and z-stack immunofluorescence. In parallel, the bacterial abundance in KP + CA was similar to KP, but the fungal abundance was higher than CA (culture method), implying that CA grows better in the presence of KP. Proteomic analysis was performed to compare KP + CA biofilm to KP biofilm alone. With isolated mouse neutrophils (thioglycolate induction), KP + CA biofilms induced less prominent responses than KP biofilms, as determined by (i) neutrophilic supernatant cytokines (ELISA) and (ii) neutrophil extracellular traps (NETs), using immunofluorescent images (neutrophil elastase, myeloperoxidase, and citrullinated histone 3), peptidyl arginine deiminase 4 (PAD4) expression, and cell-free DNA. Likewise, intratracheal KP + CA in C57BL/6 mice induces less severe pneumonia than KP alone, as indicated by organ injury (serum creatinine and alanine transaminase) (colorimetric assays), cytokines (ELISA), bronchoalveolar lavage fluid parameters (bacterial culture and neutrophil abundances using a hemocytometer), histology score (H&E stains), and NETs (immunofluorescence on the lung tissue). In conclusion, the biofilm biomass of KP + CA was mostly produced from CA with less potent neutrophil activation and less severe pneumonia than KP alone. Hence, fungi in the respiratory tract might benefit the host in some situations, despite the well-known adverse effects of fungi. Full article
(This article belongs to the Special Issue Mechanisms in Biofilm Formation, Tolerance and Control: 2nd Edition)
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20 pages, 32609 KB  
Article
Shaking Table Tests and Numerical Study on the Seismic Performance of Arc-Shaped Shear Keys in Highway Continuous-Girder Bridges
by Liang Huang, Wenze Wang, Shizhan Xu, Bo Wang and Zisheng Li
Buildings 2024, 14(10), 3060; https://doi.org/10.3390/buildings14103060 - 25 Sep 2024
Cited by 2 | Viewed by 1229
Abstract
Typical forms of seismic damage to laminated-rubber-bearing girder bridges in the transverse direction are falling beams, girder displacement, and bearing damage. However, the damage to piers and foundations is generally lighter. This is mainly due to slippage of the bearings. Therefore, we propose [...] Read more.
Typical forms of seismic damage to laminated-rubber-bearing girder bridges in the transverse direction are falling beams, girder displacement, and bearing damage. However, the damage to piers and foundations is generally lighter. This is mainly due to slippage of the bearings. Therefore, we propose a new type of arc-shaped shear key to improve the lateral seismic performance. A 1/12-scale highway continuous-girder bridge isolated by different shear keys was tested utilizing a 4 m × 4 m shaking table with six DOFs. The seismic responses of the bridge were analyzed in terms of phenomenon, displacement, strain, and acceleration. The main girder and pier exhibited different seismic responses because the bridge had different stops. A numerical simulation based on FEM showed that the established finite element model can well reproduce the displacement time history of the main girder and the cap girder. By analyzing the finite element model, the relative displacement of the bearing under different seismic waves was obtained. A comparison between the measured and FEM responses showed that the arc-shaped shear key can well limit the displacement of the main girder and the bearing. In addition, it does not significantly amplify the seismic response of the substructure. The arc-shaped shear key dissipates more energy while limiting the displacement of the main girder, and the comprehensive seismic performance is better than that of the rubber pad shear key. Full article
(This article belongs to the Special Issue Seismic Analysis and Design of Building Structures)
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19 pages, 16327 KB  
Article
Experimental Investigation of the Mechanical Behavior of the Strain Isolation Pad in Thermal Protection Systems under Tension
by Maoxu Lu, Zhenqiang Wu, Ziqing Hao and Liu Liu
Aerospace 2024, 11(4), 305; https://doi.org/10.3390/aerospace11040305 - 12 Apr 2024
Cited by 1 | Viewed by 1873
Abstract
A strain isolation pad is a critical connection mechanism that enables deformation coordination between the rigid thermal insulation tile and the primary structure in the thermal protection system of a reusable hypersonic vehicle. An experimental investigation has been conducted to determine the static, [...] Read more.
A strain isolation pad is a critical connection mechanism that enables deformation coordination between the rigid thermal insulation tile and the primary structure in the thermal protection system of a reusable hypersonic vehicle. An experimental investigation has been conducted to determine the static, loading–unloading, and high-cycle fatigue (HCF) responses of the SIP with 0.2 mm adhesive under through-thickness tension at room temperature. The contributions of the rigid thermal insulation tile and metallic substructure have not been considered so far. The results indicate that the tensile behavior of the SIP joint is highly nonlinear. The static and fatigue tensile failures both initiate from the corner close to the adhesive/SIP interface due to the stress concentration and the edge effect. The uniform breakage of the aramid fiber can be seen on the cross-section. A novel method is proposed to quantify the residual strain due to the short-time ratcheting effect of the SIP joint in the initial loading–unloading tensile response. As the number of fatigue cycles increases, the thickness of the SIP joint continues to increase until failure. An explicit expression associated with the growth of SIP joint thickness, fatigue cycle number, and peak cyclic stress is established. The turning point of the thickness growth rate with the fatigue cycle number is proposed as a new fatigue failure index for the SIP joint under tensile fatigue, and a fatigue life prediction model is developed. Full article
(This article belongs to the Special Issue Advanced Aerospace Composite Materials and Smart Structures)
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19 pages, 6053 KB  
Article
The Potential Teeth Bleaching and Halitosis Prevention Effects of Pediococcus inopinatus THK-30, a Kimchi-Derived Lactic Acid Bacterium: In Vitro Study
by Trang Thi Minh Nguyen, Qiwen Zheng, Eun-Ji Yi, Arce Defeo Bellere, Xiangji Jin, Hong-Yong Kim and Tae-Hoo Yi
J. Funct. Biomater. 2024, 15(3), 64; https://doi.org/10.3390/jfb15030064 - 7 Mar 2024
Cited by 2 | Viewed by 3543
Abstract
Background: Recent developments in addressing dental aesthetic concerns, encompassing issues like teeth discoloration and halitosis, underscore the demand for safer alternative solutions. Purpose: This study aims to confirm the effects of lactic acid bacteria (LAB) from kimchi on artificial teeth bleaching and their [...] Read more.
Background: Recent developments in addressing dental aesthetic concerns, encompassing issues like teeth discoloration and halitosis, underscore the demand for safer alternative solutions. Purpose: This study aims to confirm the effects of lactic acid bacteria (LAB) from kimchi on artificial teeth bleaching and their potential impact in terms of preventing halitosis-related bacteria. Materials and Methods: To evaluate the antimicrobial effects against oral pathogens, disc diffusion tests and broth microdilution methods were used. Additionally, crystal violet analysis was performed to confirm the biofilm inhibition effect. The bleaching effects on stained artificial teeth were analyzed using the CIEDE2000 colorimetric method. Statistical analyses were performed using GraphPad Prism 9 with one-way and two-way ANOVA, with the significance level set at α < 0.05. Results: The strain THK-30, isolated from kimchi, exhibited antibacterial activity against Streptococcus mutans, Porphyromonas gingivalis, and Fusobacterium nucleatum, and was identified as Pediococcus inopinatus. Moreover, THK-30 showed a synergistic antibacterial effect against Gram-negative oral pathogens with 8% sodium hexametaphosphate (SHMP). In the stained artificial teeth bleaching test and artificial teeth biofilm inhibition test, the cell-free supernatant of THK-30 displayed significant teeth bleaching effects and caused the inhibition of biofilm formation, both independently and in combination with SHMP 8%. Conclusions: This study has demonstrated the potential applicability of LAB in teeth discoloration and halitosis. These findings are poised to provide a foundation for the development of research pertaining to the control of oral bacteria. Full article
(This article belongs to the Special Issue Biomaterials in Conservative Dentistry and Prosthodontics)
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17 pages, 1457 KB  
Article
High Incidence of Multiple-Drug-Resistant Pheromone-Responsive Plasmids and Transmissions of VanA-Type Vancomycin-Resistant Enterococcus faecalis between Livestock and Humans in Taiwan
by Haruyoshi Tomita, Jang-Jih Lu and Yasuyoshi Ike
Antibiotics 2023, 12(12), 1668; https://doi.org/10.3390/antibiotics12121668 - 27 Nov 2023
Cited by 5 | Viewed by 2215
Abstract
A total of seventy VanA-type vancomycin-resistant enterococci (VRE) isolates obtained in Taiwan in the early 2000s were retrospectively characterized. Forty isolates were obtained from human patients and thirty from livestock. Of these VRE isolates, twenty-three (57.5%) of the human VRE and thirty (100%) [...] Read more.
A total of seventy VanA-type vancomycin-resistant enterococci (VRE) isolates obtained in Taiwan in the early 2000s were retrospectively characterized. Forty isolates were obtained from human patients and thirty from livestock. Of these VRE isolates, twenty-three (57.5%) of the human VRE and thirty (100%) of the livestock VRE were Enterococcus faecalis, and the remaining seventeen (42.5%) of the human VRE were E. faecium. Of the 53 E. faecalis isolates, twenty-two (96%) of the human VRE and thirty (100%) of the livestock VRE exhibited a high level of resistance to vancomycin and sensitivity to teicoplanin. They also had three amino acid substitutions in the N-terminal region of the deduced VanS sequence. The vancomycin resistance of all of the 22 human isolates, and 20 of the 30 livestock isolates, transferred to E. faecalis FA2-2 at a frequency of 10−5 to 10−3 per donor cell in broth. Each of the transconjugants responded to E. faecalis pheromone (i.e., E. faecalis FA2-2 culture filtrate), indicating that the conjugative plasmids were pheromone-responsive plasmids. Three of the conjugative plasmids originated from human isolates, and five plasmids from livestock isolates were corresponded and classified as type A plasmid. Two plasmids originated from human isolates and six plasmids from livestock isolates were corresponded and classified as type B plasmid. E. faecalis FA2-2 containing either the type A or type B plasmid responded to the synthetic pheromone cAD1. The type A and type B plasmids transferred between E. faecalis FA2-2 and JH2SS at a frequency of about 10−2 per donor cell and conferred vancomycin, bacitracin, and erythromycin resistances. The complete DNA sequence of the representative type A plasmid pTW9 (85,068 bp) showed that the plasmid carried a Tn1546-like element encoding vanA-type resistance, erythromycin resistance (ermB), and bacitracin resistance (bcrABDR). The plasmid contained the regulatory region found in the pheromone-responsive plasmid and encoded the genes traA, traD and iad1, which are the key negative regulatory elements, and traE1, a key positive regulator of plasmid pAD1, indicating that plasmid pTW9 was pAD1-type pheromone-responsive plasmid. PFGE analysis of SmaI-digested chromosomal DNAs showed that several E. faecalis strains harboring an identical type A pheromone-responsive plasmid were indistinguishable, and that these were identified both in human and livestock isolates, indicating the transmissions of the VRE strains between livestock and humans. These data showed that the multiple-drug-resistant pheromone-responsive conjugative plasmids have been widely spread in both human and livestock VRE, and there was high potential for transfers of VRE from food animals to humans in Taiwan in the early 2000s. Full article
(This article belongs to the Special Issue Antimicrobial Resistance in Foodborne Pathogens)
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15 pages, 3926 KB  
Article
Cellulose/Zeolitic Imidazolate Framework (ZIF-8) Composites with Antibacterial Properties for the Management of Wound Infections
by Valentina Di Matteo, Maria Francesca Di Filippo, Barbara Ballarin, Giovanna Angela Gentilomi, Francesca Bonvicini, Silvia Panzavolta and Maria Cristina Cassani
J. Funct. Biomater. 2023, 14(9), 472; https://doi.org/10.3390/jfb14090472 - 13 Sep 2023
Cited by 15 | Viewed by 4012
Abstract
Metal–organic frameworks (MOFs) are a class of crystalline porous materials with outstanding physical and chemical properties that make them suitable candidates in many fields, such as catalysis, sensing, energy production, and drug delivery. By combining MOFs with polymeric substrates, advanced functional materials are [...] Read more.
Metal–organic frameworks (MOFs) are a class of crystalline porous materials with outstanding physical and chemical properties that make them suitable candidates in many fields, such as catalysis, sensing, energy production, and drug delivery. By combining MOFs with polymeric substrates, advanced functional materials are devised with excellent potential for biomedical applications. In this research, Zeolitic Imidazolate Framework 8 (ZIF-8), a zinc-based MOF, was selected together with cellulose, an almost inexhaustible polymeric raw material produced by nature, to prepare cellulose/ZIF-8 composite flat sheets via an in-situ growing single-step method in aqueous media. The composite materials were characterized by several techniques (IR, XRD, SEM, TGA, ICP, and BET) and their antibacterial activity as well as their biocompatibility in a mammalian model system were investigated. The cellulose/ZIF-8 samples remarkably inhibited the growth of Gram-positive and Gram-negative reference strains, and, notably, they proved to be effective against clinical isolates of Staphylococcus epidermidis and Pseudomonas aeruginosa presenting different antibiotic resistance profiles. As these pathogens are of primary importance in skin diseases and in the delayed healing of wounds, and the cellulose/ZIF-8 composites met the requirements of biological safety, the herein materials reveal a great potential for use as gauze pads in the management of wound infections. Full article
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11 pages, 2012 KB  
Article
Metronidazole Potentiation by Panax Ginseng and Symphytum officinale: A New Strategy for P. gingivalis Infection Control
by Salah M. Ibrahim, Abbas S. Al-Mizraqchi and Julfikar Haider
Antibiotics 2023, 12(8), 1288; https://doi.org/10.3390/antibiotics12081288 - 4 Aug 2023
Cited by 14 | Viewed by 2146
Abstract
The important periodontal disease pathogen Porphyromonas gingivalis produces thick biofilms that increase its pathogenicity. Finding natural antimicrobial agents is crucial because of the rise in antibiotic resistance. The purpose of this study was to determine if plant extracts such as Symphytum officinale (S) [...] Read more.
The important periodontal disease pathogen Porphyromonas gingivalis produces thick biofilms that increase its pathogenicity. Finding natural antimicrobial agents is crucial because of the rise in antibiotic resistance. The purpose of this study was to determine if plant extracts such as Symphytum officinale (S) and Panax Ginseng (G) were effective against P. gingivalis separately and in combination with a common antibiotic, metronidazole (F). Six different dilutions were produced using the plant extracts in different concentrations and antibiotics separately and in combination with F, G, and S using the two-fold serial dilution technique. To evaluate the effects of these substances, biofilm inhibition experiments were conducted. Plaque samples were collected from periodontitis patients to isolate P. gingivalis, and a standard strain of P. gingivalis (ATCC 33277) was purchased. Additionally, Acylated Homoserine Lactones (AHLs) detection was carried out to look for any activity that would interfere with quorum sensing. GraphPad Prism was used for statistical analysis with a p-value < 0.05. The combinations of Symphytum officinale and metronidazole (S+F) showed the maximum effectiveness in biofilm inhibition (98.7%), which was slightly better than G+F (98.2%), with substantial variations in biofilm inhibition levels in different treatment regimes. Notably, the patient isolate was more active than the standard strain. Additionally, the plant extracts and their combinations at particular dilutions had notable inhibitory effects on the generation of AHL (p < 0.05). The study highlights the possibility of Symphytum officinale and Panax Ginseng as effective treatments for P. gingivalis biofilm and AHLs, both on their own and in combination with metronidazole. These organic substances may open the door to cutting-edge methods of treating periodontal disorders. Full article
(This article belongs to the Section Novel Antimicrobial Agents)
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30 pages, 5986 KB  
Article
Screening and Application of Novel Homofermentative Lactic Acid Bacteria Results in Low-FODMAP Whole-Wheat Bread
by Małgorzata Borowska, Lilit Ispiryan, Emma Neylon, Aylin W. Sahin, Craig P. Murphy, Emanuele Zannini, Elke K. Arendt and Aidan Coffey
Fermentation 2023, 9(4), 336; https://doi.org/10.3390/fermentation9040336 - 28 Mar 2023
Cited by 7 | Viewed by 4835
Abstract
FODMAPs are fermentable oligo-, di-, monosaccharides, and polyols. The application of homofermentative lactic acid bacteria (LAB) has been investigated as a promising approach for producing low-FODMAP whole-wheat bread. The low-FODMAP diet is recommended to treat irritable bowel syndrome (IBS). Wheat flour is staple [...] Read more.
FODMAPs are fermentable oligo-, di-, monosaccharides, and polyols. The application of homofermentative lactic acid bacteria (LAB) has been investigated as a promising approach for producing low-FODMAP whole-wheat bread. The low-FODMAP diet is recommended to treat irritable bowel syndrome (IBS). Wheat flour is staple to many diets and is a significant source of fructans, which are considered FODMAPs. The reduction of fructans via sourdough fermentation, generally associated with heterofermentative lactic acid bacteria (LAB), often leads to the accumulation of other FODMAPs. A collection of 244 wild-type LAB strains was isolated from different environments and their specific FODMAP utilisation profiles established. Three homofermentative strains were selected for production of whole-wheat sourdough bread. These were Lactiplantibacillus plantarum FST1.7 (FST1.7), Lacticaseibacillus paracasei R3 (R3), and Pediococcus pentosaceus RYE106 (RYE106). Carbohydrate levels in flour, sourdoughs (before and after 48 h fermentation), and resulting breads were analysed via HPAEC-PAD and compared with whole-wheat bread leavened with baker’s yeast. While strain R3 was the most efficient in FODMAP reduction, breads produced with all three test strains had FODMAP content below cut-off levels that would trigger IBS symptoms. Results of this study highlighted the potential of homofermentative LAB in producing low-FODMAP whole-wheat bread. Full article
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11 pages, 2149 KB  
Article
First Isolation and Identification of Homologous Recombination Events of Porcine Adenovirus from Wild Boar
by Mami Oba, Sumiya Borjigin, Fuka Kikuchi, Toru Oi, Hitoshi Takemae, Hiroho Ishida, Hironobu Murakami, Naoyuki Aihara, Takanori Shiga, Junichi Kamiie, Tetsuya Mizutani and Makoto Nagai
Viruses 2022, 14(11), 2400; https://doi.org/10.3390/v14112400 - 29 Oct 2022
Cited by 1 | Viewed by 2046
Abstract
Porcine adenoviruses (PAdVs) are distributed in pig populations and classified into five immunologically distinct serotypes (PAdV-1 to 5). In this study, a PAdV was isolated from a fecal sample of wild boar for the first time. Whole-genome analysis revealed that this strain (Ino5) [...] Read more.
Porcine adenoviruses (PAdVs) are distributed in pig populations and classified into five immunologically distinct serotypes (PAdV-1 to 5). In this study, a PAdV was isolated from a fecal sample of wild boar for the first time. Whole-genome analysis revealed that this strain (Ino5) has sequence homology (approximately > 93%) throughout the genome with the PAdV-5 strain HNF-70 that was isolated from a pig in Japan in 1987, except for the hexon, E3 612R, and fiber coding regions. Two possible recombination breakpoints were detected in the hexon and E3 612R regions, which were found to have reduced GC content. Structural prediction analysis showed that a part of the hexon protein corresponding to the tower region of Ino5 had structural differences when compared with HNF-70, suggesting antigenic heterogeneity between these strains. PAdVs were detected in 1.77% (2/113) and 12% (12/100) of the fecal samples from wild boars and pigs collected in Japan by PCR, respectively. Phylogenetic analyses of the hexon and fiber genes revealed that some samples showed different grouping in the hexon and fiber genes, suggesting that these viruses have recombination events. These findings suggest that the PAdV-5 has evolved with homologous recombination events in the same manner as human adenoviruses among not only pig populations, but also wild boars in Japan. Full article
(This article belongs to the Section Animal Viruses)
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10 pages, 2996 KB  
Communication
High Rates of Detection and Molecular Characterization of Porcine Adenovirus Serotype 5 (Porcine mastadenovirus C) from Diarrheic Pigs
by Kerry Gainor, Yussaira Castillo Fortuna, Angeline Steny Alakkaparambil, Wendy González, Yashpal Singh Malik and Souvik Ghosh
Pathogens 2022, 11(10), 1210; https://doi.org/10.3390/pathogens11101210 - 20 Oct 2022
Cited by 6 | Viewed by 2504
Abstract
Since the first report on isolation of porcine adenovirus serotype 5 (PAdV-5, species Porcine mastadenovirus C (PAdV-C)) from pigs with respiratory illness in Japan in 1987, PAdV-5 have been detected in a few fecal samples from healthy pigs and in some [...] Read more.
Since the first report on isolation of porcine adenovirus serotype 5 (PAdV-5, species Porcine mastadenovirus C (PAdV-C)) from pigs with respiratory illness in Japan in 1987, PAdV-5 have been detected in a few fecal samples from healthy pigs and in some environmental samples. To date, only a single PAdV-5 strain (isolate HNF-70 from 1987) has been analyzed for the complete genome. We report here high detection rates of PAdV-5 (25.74%, 26/101 fecal samples) in diarrheic pigs at 3 different farms in the Caribbean country of Dominican Republic. After a long gap, the complete deduced amino acid sequences of the DNA-dependent DNA polymerase (pol) and hexon of two PAdV-5 strains (GES7 and Z11) were determined, revealing >99% sequence identities between PAdV-5 strains (HNF-70, GES7 and Z11) detected in different parts of the world and during different time periods (1987, and 2020–2021). By phylogenetic analysis, the putative hexon and pol of HNF-70, GES7 and Z11 exhibited similar clustering patterns, with the PAdV-5 strains forming a tight cluster near ruminant AdVs, distinct from the species PAdV-A and -B. GES7 and Z11 retained the various conserved features present in the putative pol and major late promoter region of HNF-70. Considering the paucity of data on current epidemiological status and genetic diversity of PAdV in porcine populations, our findings warrant similar studies on PAdV-5 and other PAdVs in clinically ill and healthy pigs. To our knowledge, this is the first report on detection and molecular characterization of PAdV-5 (PAdV-C) from diarrheic pigs. Full article
(This article belongs to the Special Issue 10th Anniversary of Pathogens—Feature Papers)
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13 pages, 2867 KB  
Article
Antibacterial and Antifungal Silver Nanoparticles with Tunable Size Embedded in Various Cellulose-Based Matrices
by Gabriela Biliuta, Andra-Cristina Bostănaru-Iliescu, Mihai Mareș, Carla Pavlov-Enescu, Valentin Năstasă, Olga Burduniuc and Sergiu Coseri
Molecules 2022, 27(19), 6680; https://doi.org/10.3390/molecules27196680 - 7 Oct 2022
Cited by 9 | Viewed by 2523
Abstract
The aim of this study was to synthesize silver nanoparticles (AgNPs) using cellulose derivatives and to evaluate their antimicrobial potential. As effective reducing and stabilizing agents for AgNPs, cellulose derivatives, such as hydroxypropyl cellulose (HPC), methylcellulose (MC), ethylcellulose (EC), and cellulose acetate (CA), [...] Read more.
The aim of this study was to synthesize silver nanoparticles (AgNPs) using cellulose derivatives and to evaluate their antimicrobial potential. As effective reducing and stabilizing agents for AgNPs, cellulose derivatives, such as hydroxypropyl cellulose (HPC), methylcellulose (MC), ethylcellulose (EC), and cellulose acetate (CA), were used. Their ability to reduce silver ions as well as the size of the resulting AgNPs were compared. The formation and stability of the reduced AgNPs in the solution were monitored using UV-Vis analysis. The size, morphology, and charge of the AgNPs were evaluated. We found that, when using cellulosic derivatives, AgNPs with sizes ranging from 17 to 89 nm and different stabilities were obtained. The parameters, such as size and ζ potential indicate the stability of AgNPs, with AgNPs-CA and AgNPs-HPC being considered more stable than AgNPs-EC and AgNPs-MC since they show higher ζ potential values. In addition, the AgNPs showed antimicrobial activity against all reference strains and clinical isolates. MIC values between 0.0312 and 0.125 mM had a bactericidal effect on both Gram-positive and Gram-negative bacteria. The fungicidal effect was obtained at a MIC value of 0.125 mM. These results may provide rational support in the design of medical gauze products, including gauze pads, rolls, and sponges. Full article
(This article belongs to the Section Green Chemistry)
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20 pages, 5621 KB  
Article
Bio-Guided Isolation of SARS-CoV-2 Main Protease Inhibitors from Medicinal Plants: In Vitro Assay and Molecular Dynamics
by Hossam M. Abdallah, Ali M. El-Halawany, Khaled M. Darwish, Mardi M. Algandaby, Gamal A. Mohamed, Sabrin R. M. Ibrahim, Abdulrahman E. Koshak, Sameh S. Elhady, Sana A. Fadil, Ali A. Alqarni, Ashraf B. Abdel-Naim and Mahmoud A. Elfaky
Plants 2022, 11(15), 1914; https://doi.org/10.3390/plants11151914 - 24 Jul 2022
Cited by 16 | Viewed by 3513
Abstract
Since the emergence of the pandemic of the coronavirus disease (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the discovery of antiviral phytoconstituents from medicinal plants against SARS-CoV-2 has been comprehensively researched. In this study, thirty-three plants belonging to seventeen different [...] Read more.
Since the emergence of the pandemic of the coronavirus disease (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the discovery of antiviral phytoconstituents from medicinal plants against SARS-CoV-2 has been comprehensively researched. In this study, thirty-three plants belonging to seventeen different families used traditionally in Saudi Arabia were tested in vitro for their ability to inhibit the SARS-CoV-2 main protease (MPRO). Major constituents of the bio-active extracts were isolated and tested for their inhibition potential against this enzyme; in addition, their antiviral activity against the SARS-CoV-2 Egyptian strain was assessed. Further, the thermodynamic stability of the best active compounds was studied through focused comparative insights for the active metabolites regarding ligand–target binding characteristics at the molecular level. Additionally, the obtained computational findings provided useful directions for future drug optimization and development. The results revealed that Psiadia punctulata, Aframomum melegueta, and Nigella sativa extracts showed a high percentage of inhibition of 66.4, 58.7, and 31.5%, against SARS-CoV-2 MPRO, respectively. The major isolated constituents of these plants were identified as gardenins A and B (from P. punctulata), 6-gingerol and 6-paradol (from A. melegueta), and thymoquinone (from N. sativa). These compounds are the first to be tested invitro against SARS-CoV-2 MPRO. Among the isolated compounds, only thymoquinone (THY), gardenin A (GDA), 6-gingerol (GNG), and 6-paradol (PAD) inhibited the SARS-CoV-2 MPRO enzyme with inhibition percentages of 63.21, 73.80, 65.2, and 71.8%, respectively. In vitro assessment of SARS-CoV-2 (hCoV-19/Egypt/NRC-03/2020 (accession number on GSAID: EPI_ISL_430820) revealed a strong-to-low antiviral activity of the isolated compounds. THY showed relatively high cytotoxicity and was anti-SARS-CoV-2, while PAD demonstrated a cytotoxic effect on the tested VERO cells with a selectivity index of CC50/IC50 = 1.33 and CC50/IC50 = 0.6, respectively. Moreover, GNG had moderate activity at non-cytotoxic concentrations in vitro with a selectivity index of CC50/IC50 = 101.3/43.45 = 2.3. Meanwhile, GDA showed weak activity with a selectivity index of CC50/IC50 = 246.5/83.77 = 2.9. The thermodynamic stability of top-active compounds revealed preferential stability and SARS-CoV-2 MPRO binding affinity for PAD through molecular-docking-coupled molecular dynamics simulation. The obtained results suggest the treating potential of these plants and/or their active metabolites for COVID-19. However, further in-vivo and clinical investigations are required to establish the potential preventive and treatment effectiveness of these plants and/or their bio-active compounds in COVID-19. Full article
(This article belongs to the Special Issue Biological and Chemical Activity of Metabolites of Medicinal Plants)
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