Development of drug resistance in opportunistic pathogens is one of the major healthcare challenges associated with infection management. Combination therapy has many advantages due to the simultaneous action of two drugs on two separate cellular targets. However, selection of the drugs should offer safety and synergistic interaction against most of the strains. Here, the efficacy of antibiotics in combination with quercetin, a natural flavonoid capable of targeting quorum sensing was tested against biofilm-forming Pseudomonas aeruginosa strains previously isolated from catheter associated urinary tract infection. Based on the antibiotic susceptibility pattern, synergistic effect of quercetin with selected antibiotics (levofloxacin, ceftriaxone, gentamycin, tobramycin and amikacin) was tested at the fractional concentrations of MIC by the checkerboard method and the fractional inhibitory concentration index (FICi) was calculated to estimate the synergistic effect. Effect of the synergistic combinations were further tested using time-kill assay, and against biofilm formation and biofilm cell viability. Cytotoxicity assays were performed using Human Embryonic Kidney 293T cells (HEK-293T) using the effective drug combinations with respective controls. The biofilm formation and biofilm cell viability were drastically affected with quercetin and selected antibiotics combinations with ≥80% inhibition. In vitro infection studies showed that all the strains could exert significant cell killing (68 to 85%) and the drug combinations decreased the infection rate significantly by reducing the cell killing effect of P. aeruginosa (p<0.05). The synergistic effect of quercetin is attributed to its quorum sensing inhibitory properties. These findings indicate that quercetin along with existing antibiotics can potentiate the treatment against P. aeruginosa infection and may reduce the selection pressure due to antibiotic overuse.
Background: Utilization of macroalgae has gained much attention in the field of pharmaceuticals, nutraceuticals, food and bioenergy. Macroalgae has been widely consumed in Asian countries as food from ancient days and proved that it has potential bioactive compounds which are responsible for its nutritional properties. Macroalgae consists of a diverse range of bioactive compounds including proteins, lipids, pigments, polysaccharides, etc. Polysaccharides from macroalgae have been utilized in food industries as gelling agents and drug excipients in the pharmaceutical industries owing to their biocompatibility and gel forming properties. Exploration of macroalgae derived sulfated polysaccharides in biomedical applications is increasing recently. Method: In the current review, we have provided information of three different sulfated polysaccharides such as carrageenan, fucoidan and ulvan and their isolation procedure (enzymatic precipitation, microwave assisted method, and enzymatic hydrolysis method), structural details, and their biomedical applications exclusively for bone tissue repair and regeneration. Results: From the scientific results on sulfated polysaccharides from macroalgae, we conclude that sulfated polysaccharides have exceptional properties in terms of hydrogel-forming ability, scaffold formation, and mimicking the extracellular matrix, increasing alkaline phosphatase activity, enhancement of biomineralization ability and stem cell differentiation for bone tissue regeneration. Conclusion: Overall, sulfated polysaccharides from macroalgae may be promising biomaterials in bone tissue repair and regeneration.
A novel Gram-stain-negative, horseshoe-shaped, non-motile bacterium, designated strain M12-11B T , was isolated from a marine sediment sample collected at a depth of 200 m from Kongsfjorden, Svalbard. The colony colour was orangish red due to the presence of carotenoids. Fatty acids were dominated by branched and unsaturated fatty acids (90.8 %), with a high abundance of iso-C 15 : 0 (14.9 %), anteiso-C 15 : 0 (11.4 %), iso-C 15 : 1 G (13.1 %), C 15 : 1 v6c (5.4 %), C 17 : 1 v6c (6.7 %), summed feature 3 (C 16 : 1 v7c and/or C 16 : 1 v6c; 9.3 %) and summed feature 9 (10-methyl C 16 : 0 and/or iso-C 17 : 1 v9c; 5.9 %). Strain M12-11B T contained MK-7 as the major respiratory quinone. The polar lipids consisted of phosphatidylcholine, phosphatidylethanolamine, one unidentified aminolipid and three unidentified lipids. Based on 16S rRNA gene sequence similarities, the type strains of Cyclobacterium amurskyense, Cyclobacterium marinum and Cyclobacterium lianum were most closely related to M12-11B T with sequence similarities of 98.2, 96.8 and 93.3 %, respectively. Other members of the family Cyclobacteriaceae had sequence similarities of ,92.0 %. However, DNA-DNA hybridization with Cyclobacterium amurskyense KCTC 12363 T and Cyclobacterium marinum DSM 745 T showed relatedness values of only 24.5 and 32.5 % with respect to strain M12-11B T . Based on the results of DNA-DNA hybridization experiments and phenotypic and chemotaxonomic data, it appears that strain M12-11B T represents a novel species of the genus Cyclobacterium, for which the name Cyclobacterium qasimii sp. nov. is proposed; the type strain is M12-11B T (5KCTC 23011 T 5NBRC 106168 T ) and it has a DNA G+C content of 40.5 mol%.
Uropathogenic bacteria are widely distributed in the environment and urinary tract infection is implicated in kidney stone disease. Here, we report on a urease negative bacterium Kalamiella piersonii (strain YU22) isolated from the urine of a struvite stone (MgNH4PO4·6H2O) patient. The closest species, K. piersonii IIIF1SW-P2T was reported from International Space Station samples. However, there are no earlier reports on its human association. Using whole genome and experimental analysis, its involvement in urinary tract colonization and struvite crystallization was explored. The strain YU22 showed many virulence factors that are needed for host cell invasion and colonization including cell adhesion factors, swimming and swarming motilities, biofilm and siderophore among others. In vitro infection studies in HEK-293T cells demonstrated the host cell attachment and killing. It was able to utilize amino acids as sole carbon source and showed growth in synthetic and healthy urine establishing metabolic adaptation to urinary tract. Increased pH and availability of ammonium ions from amino acid breakdown promoted struvite crystallization. The results from this study support the involvement of urease negative uropathogen in the struvite lithogenesis. Further studies on other isolates of K. peirsonii are warranted to assess its health risks.
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