Purification and characterization of deoxyribonuclease from small intestine of camel Camelus dromedarius

Abdel-Gany, Somia S.; El-Badry, Mohamed O.; Fahmy, Afaf S.; Mohamed, Saleh A.

doi:10.1016/j.jgeb.2017.06.008

Cited by 5 publications

(4 citation statements)

References 33 publications

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“…Both enzymes are proved to be stable up to 50 °C. 23 , 24 Further sonication was performed by a probe-type sonicator at 35 W for 30 min to produce the final product. The scheme of the liposome preparation procedures is illustrated in Suppl.…”

Section: Methodsmentioning

confidence: 99%

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Facile Biofilm Penetration of Cationic Liposomes Loaded with DNase I/Proteinase K to Eradicate Cutibacterium acnes for Treating Cutaneous and Catheter Infections

Fang¹,

Chou²,

Lin³

et al. 2021

IJN

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Background The biofilm produced by Cutibacterium acnes is a major infection threat for skin and implanted catheters. Nanoparticles provide a new approach to eradicate biofilms. The present study evaluated the capability of cationic liposomes loaded with DNase I (DNS) and proteinase K (PK) to remove preformed C. acnes biofilms. Methods DNS and PK were able to target and disassemble the biofilm by degrading extracellular polymer substances (EPS). Soyaethyl morpholinium ethosulfate (SME) was used to render a positive charge and enhance the antibacterial activity of the liposomes. Results The cationic liposomes containing enzymes yielded monodisperse nanovesicles ranging between 95 and 150 nm. The entrapment efficiency of the enzymes in the liposomes achieved a value of 67–83%. All liposomal formulations suppressed planktonic C. acnes growth at a minimum inhibitory concentration (MIC) equal to the free SME in the solution. The enzyme in the liposomal form inhibited biofilm growth much better than that in the free form, with the dual enzyme-loaded liposomes demonstrating the greatest inhibition of 54% based on a crystal violet assay. The biofilm-related virulence genes PA380 and PA1035 were downregulated by the combined enzymes in the liposomes but not the individual DNS or PK. Scanning electron microscopy (SEM) and confocal microscopy displayed reduced C. acnes aggregates and biofilm thickness by the liposomal system. The liposomes could penetrate through about 85% of the biofilm thickness. The in vitro pig skin permeation also showed a facile delivery of liposomes into the epidermis, deeper skin strata, and hair follicles. The liposomes exhibited potent activity to eliminate C. acnes colonization in mouse skin and catheters in vivo. The colony-forming units (CFUs) in the catheter treated with the liposomes were reduced by 2 logs compared to the untreated control. Conclusion The data suggested a safe application of the enzyme-loaded cationic liposomes as antibacterial and antibiofilm agents.

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Section: Methodsmentioning

confidence: 99%

“…Hematoxylin and eosin (H&E) staining was performed for a histological examination of the skin under optical microscopy, as described previously. 24 …”

Section: Methodsmentioning

confidence: 99%

Facile Biofilm Penetration of Cationic Liposomes Loaded with DNase I/Proteinase K to Eradicate Cutibacterium acnes for Treating Cutaneous and Catheter Infections

Fang¹,

Chou²,

Lin³

et al. 2021

IJN

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“…While we used abiotic environmental factors to quantify and predict eoDNA degradation, the actual degradation is most likely biotic, caused by free DNAses and microbes in the feces (Dash & Das, 2018; Regnaut et al, 2005). Free DNAses profit from water availability under high humidity and speed up enzymatic processes under higher temperature (Abdel‐Gany et al, 2017). Better growth of DNA‐degrading microbes at higher temperatures (Eichmiller et al, 2016) further expedites DNA degradation, for example, in feces (Murphy et al, 2007; Nsubuga et al, 2004), feathers (Vili et al, 2013), and hair (Sawaya et al, 2015).…”

Section: Discussionmentioning

confidence: 99%

How to quantify factors degrading DNA in the environment and predict degradation for effective sampling design

et al. 2023

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Extra-organismal DNA (eoDNA) from material left behind by organisms (noninvasive DNA, e.g., feces, hair) or from environmental samples (eDNA, e.g., water, soil) is a valuable source of genetic information. However, the relatively low quality and quantity of eoDNA, which can be further degraded by environmental factors, results in reduced amplification and sequencing success. This is often compensated for through cost-and time-intensive replications of genotyping/sequencing procedures.Therefore, system-and site-specific quantifications of environmental degradation are needed to maximize sampling efficiency (e.g., fewer replicates, shorter sampling durations), and to improve species detection and abundance estimates. Using 10 environmentally diverse bat roosts as a case study, we developed a robust modeling pipeline to quantify the environmental factors degrading eoDNA, predict eoDNA quality, and estimate sampling-site-specific ideal exposure duration. Maximum humidity was the strongest eoDNA-degrading factor, followed by exposure duration and then maximum temperature. We also found a positive effect when hottest days occurred later.The strength of this effect fell between the strength of the effects of exposure duration and maximum temperature. With those predictors and information on sampling period (before or after offspring were born), we reliably predicted mean eoDNA quality per sampling visit at new sites with a mean squared error of 0.0349. Site-specific simulations revealed that reducing exposure duration to 2-8 days could substantially improve eoDNA quality for future sampling. Our pipeline identified high humidity and temperature as strong drivers of eoDNA degradation even in the absence of rain and direct sunlight. Furthermore, we outline the pipeline's utility for other systems and study goals, such as estimating sample age, improving eDNA-based species detection, and increasing the accuracy of abundance estimates.

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“…We observed that DNA obtained from mice samples presented a smear whereas samples in vitro contained a single DNA band. The smear from caecum and colon samples could be derived from DNAses present in the intestinal lumen that could degrade the DNA 201 . We speculated that these unspecific fragments of different sizes could be affecting to the entire protocol, because after DNA fragmentation, adaptors will be added to these unspecific fragments.…”

Section: Transposition Mutagenesis Sequencing and Analysismentioning

confidence: 99%

Characterization of Genes and Functions Required by Multidrug-resistant Enterococci to Colonize the Intestine

Duro¹

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through nutrient depletion, specifically by reducing the levels of fructose. Finally, in vivo RNA-Seq analysis of each bacterial isolate of the consortium in combination with ex vivo and in vivo assays demonstrated that a single bacterium (Olsenella sp.) could recapitulate the protective effect.Altogether, the results obtained have identified the function of specific genes required by VRE to colonize the gut. In addition, we have identified a specific mechanism by which the microbiota confers protection against VRE colonization. These results could lead to novel therapeutic approaches to prevent infections caused by this multidrug-resistant pathogen.ARN-Seq in vivo de cada aislado en combinación con los ensayos ex vivo e in vivo demostraron que una sola bacteria (Olsenella sp.) proporciona protección.En conjunto, los resultados obtenidos han identificado la función de genes específicos requeridos por ERV para colonizar el intestino. Además, hemos identificado un mecanismo mediante el cual la microbiota confiere protección. Estos resultados podrían conducir a nuevos enfoques terapéuticos para prevenir las infecciones causadas por este patógeno multiresistente a los antibióticos.En conjunt, els resultats obtinguts han identificat la funció de gens específics requerits per ERV per a colonitzar l'intestí. A més, hem identificat un mecanisme mitjançant el qual la microbiota confereix protecció. Aquests resultats podrien conduir a nous enfocaments terapèutics per a previndre les infeccions causades per aquest patogen multiresistent als antibiòtics.

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Purification and characterization of deoxyribonuclease from small intestine of camel Camelus dromedarius

Cited by 5 publications

References 33 publications

Facile Biofilm Penetration of Cationic Liposomes Loaded with DNase I/Proteinase K to Eradicate Cutibacterium acnes for Treating Cutaneous and Catheter Infections

Facile Biofilm Penetration of Cationic Liposomes Loaded with DNase I/Proteinase K to Eradicate Cutibacterium acnes for Treating Cutaneous and Catheter Infections

How to quantify factors degrading DNA in the environment and predict degradation for effective sampling design

Characterization of Genes and Functions Required by Multidrug-resistant Enterococci to Colonize the Intestine

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