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Generation of soluble, disulfide-rich JEV NS1 protein recognizable by anti-NS1 antibodies through a simplified, in vitro refolding approach

Chong HY, Leow CY, Leow CH

Int J Biol Macromol, 2021 Aug 31;185:485-493.
PMID: 34174313 DOI: 10.1016/j.ijbiomac.2021.06.146

Co-existence of Japanese Encephalitis virus (JEV) with highly homologous antigenic epitopes results in antibody-based serodiagnosis being inaccurate at detecting and distinguishing JEV from other flaviviruses. This often causes misdiagnosis and inefficient treatments of flavivirus infection. Generation of JEV NS1 protein remains a challenge as it is notably expressed in the form of inactive aggregates known as inclusion bodies using bacterial expression systems. This study evaluated two trxB and gor E. coli strains in producing soluble JEV NS1 via a cold-shock expression system. High yield of JEV NS1 inclusion bodies was produced using cold-shocked expression system. Subsequently, a simplified yet successful approach in generating soluble, active JEV NS1 protein through solubilization, purification and in vitro refolding of JEV NS1 protein from inclusion bodies was developed. A step-wise dialysis refolding approach was used to facilitate JEV NS1 refolding. The authenticity of the refolded JEV NS1 was confirmed by specific antibody binding on indirect ELISA commercial anti-NS1 antibodies which showed that the refolded JEV NS1 was highly immunoreactive. This presented approach is cost-effective, and negates the need for mammalian or insect cell expression systems in order to synthesize this JEV NS1 protein of important diagnostic and therapeutic relevance in Japanese Encephalitis disease.

Matched MeSH terms: Escherichia coli/growth & development*
Fulltext Green Synthesized Silver Nanoparticles Immobilized on Activated Carbon Nanoparticles: Antibacterial Activity Enhancement Study and Its Application on Textiles Fabrics

Wibawa PJ, Nur M, Asy'ari M, Wijanarka W, Susanto H, Sutanto H, et al.

Molecules, 2021 Jun 22;26(13).
PMID: 34206375 DOI: 10.3390/molecules26133790

This research aimed to enhance the antibacterial activity of silver nanoparticles (AgNPs) synthesized from silver nitrate (AgNO3) using aloe vera extract. It was performed by means of incorporating AgNPs on an activated carbon nanoparticle (ACNPs) under ultrasonic agitation (40 kHz, 2 × 50 watt) for 30 min in an aqueous colloidal medium. The successful AgNPs synthesis was clarified with both Ultraviolet-Visible (UV-Vis) and Fourier Transform Infrared (FTIR) spectrophotometers. The successful AgNPs-ACNPs incorporation and its particle size analysis was performed using Transmission Electron Microscope (TEM). The brown color suspension generation and UV-Vis's spectra maximum wavelength at around 480 nm confirmed the existence of AgNPs. The particle sizes of the produced AgNPs were about 5 to 10 nm in the majority number, which collectively surrounded the aloe vera extract secondary metabolites formed core-shell like nanostructure of 8.20 ± 2.05 nm in average size, while ACNPs themselves were about 20.10 ± 1.52 nm in average size formed particles cluster, and 48.00 ± 8.37 nm in average size as stacking of other particles. The antibacterial activity of the synthesized AgNPs and AgNPs-immobilized ACNPs was 57.58% and 63.64%, respectively (for E. coli); 61.25%, and 93.49%, respectively (for S. aureus). In addition, when the AgNPs-immobilized ACNPs material was coated on the cotton and polyester fabrics, the antibacterial activity of the materials changed, becoming 19.23% (cotton; E. coli), 31.73% (polyester; E. coli), 13.36% (cotton; S. aureus), 21.15% (polyester; S. aureus).

Matched MeSH terms: Escherichia coli/growth & development*
Fulltext Compositions and antimicrobial properties of binary ZnO-CuO nanocomposites encapsulated calcium and carbon from Calotropis gigantea targeted for skin pathogens

Govindasamy GA, Mydin RBSMN, Sreekantan S, Harun NH

Sci Rep, 2021 01 08;11(1):99.
PMID: 33420110 DOI: 10.1038/s41598-020-79547-w

Calotropis gigantea (C. gigantea) extract with an ecofriendly nanotechnology approach could provide promising antimicrobial activity against skin pathogens. This study investigates the antimicrobial capability of green synthesized binary ZnO-CuO nanocomposites from C. gigantea against non-MDR (Staphylococcus aureus and Escherichia coli) and MDR (Klebsiella pneumoniae, Pseudomonas aeruginosa and methicillin-resistant S. aureus) skin pathogens. Scanning electron microscopy and transmission electron microscopy revealed the size and shape of B3Z1C sample. Results of X-ray powder diffraction, energy-dispersive spectroscopy, FTIR and UV-Vis spectroscopy analyses confirmed the presence of mixed nanoparticles (i.e., zinc oxide, copper oxide, carbon and calcium) and the stabilising phytochemical agents of plant (i.e., phenol and carbonyl). Antimicrobial results showed that carbon and calcium decorated binary ZnO-CuO nanocomposites with compositions of 75 wt% of ZnO and 25 wt% CuO (B3Z1C) was a strong bactericidal agent with the MBC/MIC ratio of ≤ 4 and ≤ 2 for non-MDR and MDR pathogens, respectively. A significant non-MDR zone of inhibitions were observed for BZC by Kirby-Bauer disc-diffusion test. Further time-kill observation revealed significant fourfold reduction in non-MDR pathogen viable count after 12 h study period. Further molecular studies are needed to explain the biocidal mechanism underlying B3Z1C potential.

Matched MeSH terms: Escherichia coli/growth & development
Montmorillonite-based polyacrylamide hydrogel rings for controlled vaginal drug delivery

Sharifzadeh G, Hezaveh H, Muhamad II, Hashim S, Khairuddin N

Mater Sci Eng C Mater Biol Appl, 2020 May;110:110609.
PMID: 32204060 DOI: 10.1016/j.msec.2019.110609

Vaginal drug delivery is regarded as a promising route against women-related health issues such as unwanted pregnancies and sexually transmitted infections. However, only a very few studies have been reported on the use of hydrogel rings with low cytotoxicity for vaginal drug delivery applications. Moreover, the effect of nanoparticles on hydrogel vaginal rings has not been clearly evaluated. To overcome these challenges, we hereby developed nanocomposite hydrogel rings based on polyacrylamide-sodium carboxymethyl cellulose-montmorillonite nanoparticles in the ring-shaped aluminum mold for controlled drug delivery. The hydrogel rings were synthesized by using N,N'-methylene bisacrylamide, N,N,N',N'-tetramethyl ethylene diamine, and ammonium persulfate, as a crosslinker, accelerator, and initiator, respectively. The obtained rings were 5.5 cm in diameters and 0.5 cm in rims. Chemical structures of the nanocomposite rings were confirmed by Fourier transform infrared, and Nuclear Magnetic Resonance spectroscopies. Additionally, the swelling ratio of hydrogels was appeared to be adjusted by the introduction of nanoparticles. In vitro release experiment of methylene blue, as a hydrophilic model drug, revealed that the nanocomposite rings could not only reduce burst effect (almost more than twice), but also achieve prolonged release for 15 days in the vaginal fluid simulant which mimic the vaginal conditions at pH of almost 4.2, and a temperature of 37 °C. Importantly, the resultant hydrogel rings with or without various concentrations of montmorillonite showed low cytotoxicity toward human skin fibroblasts. Furthermore, different antibacterial activities against Escherichia coli were observed for various concentrations of montmorillonite in hydrogels. These results suggest the great potential of montmorillonite-based hydrogel rings for vaginal drug delivery.

Matched MeSH terms: Escherichia coli/growth & development*
Multi-dimensional zinc oxide (ZnO) nanoarchitectures as efficient photocatalysts: What is the fundamental factor that determines photoactivity in ZnO?

Chang JS, Strunk J, Chong MN, Poh PE, Ocon JD

J Hazard Mater, 2020 01 05;381:120958.
PMID: 31416043 DOI: 10.1016/j.jhazmat.2019.120958

While bulk zinc oxide (ZnO) is of non-toxic in nature, ZnO nanoarchitectures could potentially induce the macroscopic characteristics of oxidative, lethality and toxicity in the water environment. Here we report a systematic study through state-of-the-art controllable synthesis of multi-dimensional ZnO nanoarchitectures (i.e. 0D-nanoparticle, 1D-nanorod, 2D-nanosheet, and 3D-nanoflowers), and subsequent in-depth understanding on the fundamental factor that determines their photoactivities. The photoactivities of resultant ZnO nanoarchitectures were interpreted in terms of the photodegradation of salicylic acid as well as inactivation of Bacillus subtilis and Escherichia coli under UV-A irradiation. Photodegradation results showed that 1D-ZnO nanorods demonstrated the highest salicylic acid photodegradation efficiency (99.4%) with a rate constant of 0.0364 min-1. 1D-ZnO nanorods also exhibited the highest log reductions of B. subtilis and E. coli of 3.5 and 4.2, respectively. Through physicochemical properties standardisation, an intermittent higher k value for pore diameter (0.00097 min-1 per mm), the highest k values for crystallite size (0.00171 min-1 per nm) and specific surface area (0.00339 min-1 per m2/g) contributed to the exceptional photodegradation performance of nanorods. Whereas, the average normalised log reduction against the physicochemical properties of nanorods (i.e. low crystallite size, high specific surface area and pore diameter) caused the strongest bactericidal effect.

Matched MeSH terms: Escherichia coli/growth & development
Recent advancement of engineering microbial hosts for the biotechnological production of flavonoids

Shah FLA, Ramzi AB, Baharum SN, Noor NM, Goh HH, Leow TC, et al.

Mol Biol Rep, 2019 Dec;46(6):6647-6659.
PMID: 31535322 DOI: 10.1007/s11033-019-05066-1

Flavonoids are polyphenols that are important organic chemicals in plants. The health benefits of flavonoids that result in high commercial values make them attractive targets for large-scale production through bioengineering. Strategies such as engineering a flavonoid biosynthetic pathway in microbial hosts provide an alternative way to produce these beneficial compounds. Escherichia coli, Saccharomyces cerevisiae and Streptomyces sp. are among the expression systems used to produce recombinant products, as well as for the production of flavonoid compounds through various bioengineering approaches including clustered regularly interspaced short palindromic repeats (CRISPR)-based genome engineering and genetically encoded biosensors to detect flavonoid biosynthesis. In this study, we review the recent advances in engineering model microbial hosts as being the factory to produce targeted flavonoid compounds.

Matched MeSH terms: Escherichia coli/growth & development
Role of coaggregation in the pathogenicity and prolonged colonisation of Vibrio cholerae

Toh YS, Yeoh SL, Yap IKS, Teh CSJ, Win TT, Thong KL, et al.

Med Microbiol Immunol, 2019 Dec;208(6):793-809.
PMID: 31263955 DOI: 10.1007/s00430-019-00628-3

Cholera is an acute diarrheal illness caused by the Gram-negative bacterium Vibrio cholerae. The pathogen is known for its ability to form biofilm that confers protection against harsh environmental condition and as part of the colonisation process during infection. Coaggregation is a process that facilitates the formation of biofilm. In a preliminary in vitro study, high coaggregation index and biofilm production were found between V. cholerae with human commensals namely Escherichia coli and Enterobacter cloacae. Building upon these results, the effects of coaggregation were further evaluated using adult BALB/c mouse model. The animal study showed no significant differences in mortality and fluid accumulation ratio between treatment groups infected with V. cholerae alone and those infected with coaggregation partnership (V. cholerae with E. coli or V. cholerae with E. cloacae). However, mild inflammation was detected in both partnering pairs. Higher density of V. cholerae was recovered from faecal samples of mice co-infected with E. coli and V. cholerae in comparison with other groups at 24 h post-infection. This partnership also elicited slightly higher levels of interleukin-5 (IL-5) and interleukin-10 (IL-10). Nonetheless, the involvement of autoinducer-2 (AI-2) as the signalling molecules in quorum sensing system is not evident in this study. Since E. coli is one of the common commensals, our result may suggest the involvement of commensals in cholera development.

Matched MeSH terms: Escherichia coli/growth & development
Gellan gum incorporating titanium dioxide nanoparticles biofilm as wound dressing: Physicochemical, mechanical, antibacterial properties and wound healing studies

Ismail NA, Amin KAM, Majid FAA, Razali MH

Mater Sci Eng C Mater Biol Appl, 2019 Oct;103:109770.
PMID: 31349525 DOI: 10.1016/j.msec.2019.109770

In this work, the potential of titanium dioxide nanoparticles incorporated gellan gum (GG + TiO2-NPs) biofilm as wound dressing material was investigated. The GG + TiO2-NPs biofilm was prepared via evaporative casting technique and was characterized using FTIR, XRD, and SEM to study their physiochemical properties. The mechanical properties, swelling and water vapor transmission rate (WVTR) of biofilm was determined to comply with an ideal wound dressing material. In vitro and in vivo wound healing studies was carried out to evaluate the performance of GG + TiO2-NPs biofilm. In vitro wound healing was studied on 3 T3 mouse fibroblast cells for cell viability, cell proliferation, and scratch assay. The acridine orange/propidium iodide (AO/PI) staining and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay were used to evaluate the viability of cell and cell proliferation. Cell migration assay was analyzed using Essen BioScience IncuCyteTM Zoom system. In vivo wound healing via open excision wounds model on Sprague Dawley rat was studied within 14 days. The FT-IR spectra of GG + TiO2-NPs biofilm show main bands assigned to OH stretching, OH deformation, and TiO stretching modes. XRD pattern of GG + TiO2-NPs biofilm suggesting that TiO2-NPs was successfully incorporated in biofilm and well distributed on the surface as proved by SEM analysis. The GG + TiO2-NPs biofilm shows higher mechanical strength and swelling (3.76 ± 0.11 MPa and 1061 ± 6%) as compared to pure GG film (3.32 ± 0.08 Mpa and 902 ± 6%), respectively. GG + TiO2-NPs biofilm shows good antibacterial properties as 9 ± 0.25 mm and 11 ± 0.06 mm exhibition zone was observed against Staphylococcus aureus and Escherichia coli bacteria, respectively. While no exhibition zone was obtained for pure GG biofilm. GG + TiO2-NPs biofilm also demonstrated better cell-to-cell interaction properties, as it's promoted cell proliferation and cell migration to accelerate open excision wound healing on Sprague Dawley rat. The wound treated with GG + TiO2-NPs biofilm was healed within 14 days, on the other hand, the wound is still can be seen when it was treated with GG. However, GG and GG + TiO2-NPs biofilm show no cytotoxicity effects on mouse fibroblast cells.

Matched MeSH terms: Escherichia coli/growth & development*
Synthesis, spectral analysis and antibacterial activity of some novel 5-substituted-2-((6-bromo-3,4-methylenedioxybenzyl)thio)-1,3,4-oxadiazole derivatives

Rehman A, Siddiqa A, Abbasi MA, Siddiqui SZ, Khan SG, Rasool S, et al.

Pak J Pharm Sci, 2018 Sep;31(5):1783-1790.
PMID: 30150171

A number of novel 5-substituted-2-((6-bromo-3,4-methylenedioxybenzyl)thio)-1,3,4-Oxadiazole derivatives (6a-l) have been synthesized to evaluate their antibacterial activity. Using aryl/aralkyl carboxylic acids (1a-l) as precursors, 5-substituted-1,3,4-Oxadiazol-2-thiols (4a-l) were yielded in good amounts. The derivatives, 4a-l, were subjected to electrophilic substitution reaction on stirring with 6-bromo-3,4-methylenedioxybenzyl chloride (5) in DMF to synthesize the required compounds. All the synthesized molecules were well characterized by IR, 1H-NMR, 13C-NMR and EIMS spectral data and evaluated for antibacterial activity against some bacterial strains of Gram-bacteria. The molecule, 6d, demonstrated the best activity among all the synthesized molecules exhibiting weak to moderate inhibition potential.

Matched MeSH terms: Escherichia coli/growth & development
Fulltext Microtiter miniature shaken bioreactor system as a scale-down model for process development of production of therapeutic alpha-interferon2b by recombinant Escherichia coli

Tan JS, Abbasiliasi S, Kadkhodaei S, Tam YJ, Tang TK, Lee YY, et al.

BMC Microbiol, 2018 01 04;18(1):3.
PMID: 29439680 DOI: 10.1186/s12866-017-1145-9

BACKGROUND: Demand for high-throughput bioprocessing has dramatically increased especially in the biopharmaceutical industry because the technologies are of vital importance to process optimization and media development. This can be efficiently boosted by using microtiter plate (MTP) cultivation setup embedded into an automated liquid-handling system. The objective of this study was to establish an automated microscale method for upstream and downstream bioprocessing of α-IFN2b production by recombinant Escherichia coli. The extraction performance of α-IFN2b by osmotic shock using two different systems, automated microscale platform and manual extraction in MTP was compared.
RESULTS: The amount of α-IFN2b extracted using automated microscale platform (49.2 μg/L) was comparable to manual osmotic shock method (48.8 μg/L), but the standard deviation was 2 times lower as compared to manual osmotic shock method. Fermentation parameters in MTP involving inoculum size, agitation speed, working volume and induction profiling revealed that the fermentation conditions for the highest production of α-IFN2b (85.5 μg/L) was attained at inoculum size of 8%, working volume of 40% and agitation speed of 1000 rpm with induction at 4 h after the inoculation.
CONCLUSION: Although the findings at MTP scale did not show perfect scalable results as compared to shake flask culture, but microscale technique development would serve as a convenient and low-cost solution in process optimization for recombinant protein.

Matched MeSH terms: Escherichia coli/growth & development
Fulltext Composition and Bioactivity of Essential Oil from Citrus grandis (L.) Osbeck 'Mato Peiyu' Leaf

Tsai ML, Lin CD, Khoo KA, Wang MY, Kuan TK, Lin WC, et al.

Molecules, 2017 Dec 05;22(12).
PMID: 29206180 DOI: 10.3390/molecules22122154

'Mato Peiyu' pomelo (Citrus grandis (L.) Osbeck 'Mato Peiyu') leaves from pruning are currently an agricultural waste. The aim of this study was to isolate essential oils from these leaves through steam distillation (SD) and solvent-free microwave extraction (SFME) and to evaluate their applicability to skin care by analyzing their antimicrobial, antioxidant (diphenyl-1-picrylhydrazyl scavenging assay, β-carotene/linoleic acid assay, and nitric oxide scavenging assay), anti-inflammatory (5-lipoxygenase inhibition assay), and antityrosinase activities. The gas chromatography-mass spectrometry results indicated that the main components of 'Mato Peiyu' leaf essential oils were citronellal and citronellol, with a total percentage of 50.71% and 59.82% for SD and SFME, respectively. The highest bioactivity among all assays was obtained for 5-lipoxygenase inhibition, with an IC50 value of 0.034% (v/v). The MIC90 of the antimicrobial activity of essential oils against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Candida albicans ranged from 0.086% to 0.121% (v/v). Citronellal and citronellol were the main contributors, accounting for at least 54.58% of the essential oil's bioactivity. This paper is the first to report the compositions and bioactivities of 'Mato Peiyu' leaf essential oil, and the results imply that the pomelo leaf essential oil may be applied in skin care.

Matched MeSH terms: Escherichia coli/growth & development
Modifications in the pmrB gene are the primary mechanism for the development of chromosomally encoded resistance to polymyxins in uropathogenic Escherichia coli

Phan MD, Nhu NTK, Achard MES, Forde BM, Hong KW, Chong TM, et al.

J Antimicrob Chemother, 2017 10 01;72(10):2729-2736.
PMID: 29091192 DOI: 10.1093/jac/dkx204

Objectives: Polymyxins remain one of the last-resort drugs to treat infections caused by MDR Gram-negative pathogens. Here, we determined the mechanisms by which chromosomally encoded resistance to colistin and polymyxin B can arise in the MDR uropathogenic Escherichia coli ST131 reference strain EC958.
Methods: Two complementary approaches, saturated transposon mutagenesis and spontaneous mutation induction with high concentrations of colistin and polymyxin B, were employed to select for mutations associated with resistance to polymyxins. Mutants were identified using transposon-directed insertion-site sequencing or Illumina WGS. A resistance phenotype was confirmed by MIC and further investigated using RT-PCR. Competitive growth assays were used to measure fitness cost.
Results: A transposon insertion at nucleotide 41 of the pmrB gene (EC958pmrB41-Tn5) enhanced its transcript level, resulting in a 64- and 32-fold increased MIC of colistin and polymyxin B, respectively. Three spontaneous mutations, also located within the pmrB gene, conferred resistance to both colistin and polymyxin B with a corresponding increase in transcription of the pmrCAB genes. All three mutations incurred a fitness cost in the absence of colistin and polymyxin B.
Conclusions: This study identified the pmrB gene as the main chromosomal target for induction of colistin and polymyxin B resistance in E. coli.

Matched MeSH terms: Uropathogenic Escherichia coli/growth & development
Preparation and properties of cellulose nanocomposite films with in situ generated copper nanoparticles using Terminalia catappa leaf extract

Muthulakshmi L, Rajini N, Nellaiah H, Kathiresan T, Jawaid M, Rajulu AV

Int J Biol Macromol, 2017 Feb;95:1064-1071.
PMID: 27984140 DOI: 10.1016/j.ijbiomac.2016.09.114

In the present work, copper nanoparticles (CuNPs) were in situ generated inside cellulose matrix using Terminalia catappa leaf extract as a reducing agent. During this process, some CuNPs were also formed outside the matrix. The CuNPs formed outside the matrix were observed with transmission electron microscope (TEM) and scanning electron microscope (SEM). Majority of the CuNPs formed outside the matrix were in the size range of 21-30nm. The cellulose/CuNP composite films were characterized by Fourier transform infrared spectroscopic, X-Ray diffraction and thermogravimetric techniques. The crystallinity of the cellulose/CuNP composite films was found to be lower than that of the matrix indicating rearrangement of cellulose molecules by in situ generated CuNPs. Further, the expanded diffractogram of the composite films indicated the presence of a mixture of Cu, CuO and Cu2O nanoparticles. The thermal stability of the composites was found to be lower than that of the composites upto 350°C beyond which a reverse trend was observed. This was attributed to the catalytic behaviour of CuNPs for early degradation of the composites. The composite films possessed sufficient tensile strength which can replace polymer packaging films like polyethylene. Further, the cellulose/CuNP composite films exhibited good antibacterial activity against E.coli bacteria.

Matched MeSH terms: Escherichia coli/growth & development
Occurrence and distribution of bacteria indicators, chemical tracers and pathogenic vibrios in Singapore coastal waters

Goh SG, Bayen S, Burger D, Kelly BC, Han P, Babovic V, et al.

Mar Pollut Bull, 2017 Jan 15;114(1):627-634.
PMID: 27712861 DOI: 10.1016/j.marpolbul.2016.09.036

Water quality in Singapore's coastal area was evaluated with microbial indicators, pathogenic vibrios, chemical tracers and physico-chemical parameters. Sampling sites were grouped into two clusters (coastal sites at (i) northern and (ii) southern part of Singapore). The coastal sites located at northern part of Singapore along the Johor Straits exhibited greater pollution. Principal component analysis revealed that sampling sites at Johor Straits have greater loading on carbamazepine, while turbidity poses greater influence on sampling sites at Singapore Straits. Detection of pathogenic vibrios was also more prominent at Johor Straits than the Singapore Straits. This study examined the spatial variations in Singapore's coastal water quality and provided the baseline information for health risk assessment and future pollution management.

Matched MeSH terms: Escherichia coli/growth & development
Effect of zinc oxide amounts on the properties and antibacterial activities of zeolite/zinc oxide nanocomposite

Alswat AA, Ahmad MB, Saleh TA, Hussein MZB, Ibrahim NA

Mater Sci Eng C Mater Biol Appl, 2016 Nov 01;68:505-511.
PMID: 27524047 DOI: 10.1016/j.msec.2016.06.028

Nanocomposites of zinc oxide loaded on a zeolite (Zeolite/ZnO NCs) were prepared using co-precipitation method. The ratio effect of ZnO wt.% to the Zeolite on the antibacterial activities was investigated. Various techniques were used for the nanocomposite characterization, including UV-vis, FTIR, XRD, EDX, FESEM and TEM. XRD patterns showed that ZnO peak intensity increased while the intensities of Zeolite peaks decreased. TEM images indicated a good distribution of ZnO-NPs onto the Zeolite framework and the cubic structure of the zeolite was maintained. The average particle size of ZnO-nanoparticles loaded on the surface of the Zeolite was in the range of 1-10nm. Moreover, Zeolite/ZnO NCs showed noticeable antibacterial activities against the tested bacteria; Gram- positive and Gram- negative bacteria, under normal light. The efficiency of the antibacterial increased with increasing the wt.% from 3 to 8 of ZnO NPs, and it reached 87% against Escherichia coli E266.

Matched MeSH terms: Escherichia coli/growth & development*
Expression of soluble human Neonatal Fc-receptor (FcRn) in Escherichia coli through modification of growth environment

Ng WK, Lim TS, Lai NS

Protein Expr. Purif., 2016 11;127:73-80.
PMID: 27412717 DOI: 10.1016/j.pep.2016.07.004

Neonatal Fc-receptor (FcRn) with its affinity to immunoglobulin G (IgG) has been the subject of many pharmacokinetic studies in the past century. This protein is well known for its unique feature in maintaining the circulating IgG from degradation in blood plasma. FcRn is formed by non-covalent association between the α-chain with the β-2-microglobulin (β2m). Many studies have been conducted to produce FcRn in the laboratory, mainly using mammalian tissue culture as host for recombinant protein expression. In this study, we demonstrate a novel strategy to express the α-chain of FcRn using Escherichia coli as the expression host. The expression vector that carries the cDNA of the α-chain was transformed into expression host, Rosetta-gami 2 strain for inducible expression. The bacterial culture was grown in a modified growth medium which constitutes of terrific broth, sodium chloride (NaCl), glucose and betaine. A brief heat shock at 45 °C was carried out after induction, before the temperature for expression was reduced to 22 °C and grown for 16 h. The soluble form of the α-chain of FcRn expressed was tested in the ELISA and dot blot immunoassay to confirm its native functionality. The results implied that the α-chain of FcRn expressed using this method is functional and retains its pH-dependent affinity to IgG. Our study significantly suggests that the activity of human FcRn remain active and functional in the absence of β2m.

Matched MeSH terms: Escherichia coli/growth & development*
Plasma polymerized carvone as an antibacterial and biocompatible coating

Chan YW, Siow KS, Ng PY, Gires U, Yeop Majlis B

Mater Sci Eng C Mater Biol Appl, 2016 Nov 01;68:861-871.
PMID: 27524089 DOI: 10.1016/j.msec.2016.07.040

Antibacterial coating is important to prevent the colonization of medical devices by biofilm forming bacteria that would cause infection and sepsis in patients. Current coating techniques such as immobilization of antimicrobial compounds, time-releasing antibiotic agents and silver nanoparticles, require multiple processing steps, and they have low efficacy and low stability. We proposed a single-step plasma polymerization of an essential oil known as carvone to produce a moderately hydrophobic antibacterial coating (ppCar) with an average roughness of <1nm. ppCar had a static water contact angle of 78°, even after 10days of air aging and it maintained its stability throughout 24h of LB broth immersion. ppCar showed promising results in the live-dead fluorescence assay and crystal violet assay. The biofilm assay showed an effective reduction of E. coli and S. aureus bacteria by 86% and 84% respectively. ppCar is also shown to rupture the bacteria membrane for its bactericidal effects. The cytotoxicity test indicated that the coating is not cytotoxic to the human cell line. This study would be of interest to researcher keen on producing a bacteria-resistance and biocompatible coating on different substrates in a cost-effective manner.

Matched MeSH terms: Escherichia coli/growth & development*
Fulltext Risk factors and spatial distribution of extended spectrum β-lactamase-producing- Escherichia coli at retail poultry meat markets in Malaysia: a cross-sectional study

Aliyu AB, Saleha AA, Jalila A, Zunita Z

BMC Public Health, 2016 08 02;16:699.
PMID: 27484086 DOI: 10.1186/s12889-016-3377-2

BACKGROUND: The significant role of retail poultry meat as an important exposure pathway for the acquisition and transmission of extended spectrum β-lactamase-producing Escherichia coli (ESBL-EC) into the human population warrants understanding concerning those operational practices associated with dissemination of ESBL-EC in poultry meat retailing. Hence, the objective of this study was to determine the prevalence, spatial distribution and potential risk factors associated with the dissemination of ESBL-EC in poultry meat retail at wet-markets in Selangor, Malaysia.
METHODS: Poultry meat (breast, wing, thigh, and keel) as well as the contact surfaces of weighing scales and cutting boards were sampled to detect ESBL-EC by using culture and disk combination methods and polymerase chain reaction assays. Besides, questionnaire was used to obtain data and information pertaining to those operational practices that may possibly explain the occurrence of ESBL-EC. The data were analysed using logistic regression analysis at 95 % CI.
RESULTS: The overall prevalence of ESBL-EC was 48.8 % (95 % CI, 42 - 55 %). Among the risk factors that were explored, type of countertop, sanitation of the stall environment, source of cleaning water, and type of cutting board were found to be significantly associated with the presence of ESBL-EC.
CONCLUSIONS: Thus, in order to prevent or reduce the presence of ESBL-EC and other contaminants at the retail-outlet, there is a need to design a process control system based on the current prevailing practices in order to reduce cross contamination, as well as to improve food safety and consumer health.

Matched MeSH terms: Escherichia coli/growth & development*
Functionally graded polymeric materials: A brif review of current fabrication methods and introduction of a novel fabrication method

Almasi D, Sadeghi M, Lau WJ, Roozbahani F, Iqbal N

Mater Sci Eng C Mater Biol Appl, 2016 Jul 01;64:102-107.
PMID: 27127033 DOI: 10.1016/j.msec.2016.03.053

The present work reviews the current fabrication methods of the functionally graded polymeric material (FGPM) and introduces a novel fabrication method that is versatile in applications as compared to those of existing used methods. For the first time electrophoresis was used to control the distribution of the tetracycline hydrochloride (TC) in a film made of polylactic acid (PLA), aiming to induce antimicrobial effect on the film prepared. The elemental analysis on the film surface showed that by employing electrophoresis force, higher amount of TC was detected near the top surface of the film. Results also showed that the FGPM samples with higher percentage of the TC on the film surface were highly effective to minimize the growth of Escherichia coli. These findings are useful and important to improve dispersion quality of the particles in the composite material and further enhance its antibacterial property.

Matched MeSH terms: Escherichia coli/growth & development*
Characterization and antibacterial activity of silver exchanged regenerated NaY zeolite from surfactant-modified NaY zeolite

Salim MM, Malek NANN

Mater Sci Eng C Mater Biol Appl, 2016 Feb;59:70-77.
PMID: 26652350 DOI: 10.1016/j.msec.2015.09.099

The antibacterial activity of regenerated NaY zeolite (thermal treatment from cetyltrimethyl ammonium bromide (CTAB)-modified NaY zeolite and pretreatment with Na ions) loaded with silver ions were examined using the broth dilution minimum inhibitory concentration (MIC) method against Escherichia coli (E. coli ATCC 11229) and Staphylococcus aureus (S. aureus ATCC 6538). X-ray diffraction (XRD), attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy, field emission scanning electron microscopy (FESEM), energy dispersive X-ray (EDX) and chemical elemental analyses were used to characterize the regenerated NaY and AgY zeolites. The XRD patterns indicated that the calcination and addition of silver ions on regenerated NaY zeolite did not affect the structure of the regenerated NaY zeolite as the characteristic peaks of the NaY zeolite were retained, and no new peaks were observed. The regenerated AgY zeolite showed good antibacterial activity against both bacteria strains in distilled water, and the antibacterial activity of the samples increased with increasing Ag loaded on the regenerated AgY zeolite; the regenerated AgY zeolite was more effective against E. coli than S. aureus. However, the antibacterial activity of the regenerated AgY was not effective in saline solution for both bacteria. The study showed that CTAB-modified NaY zeolite materials could be regenerated to NaY zeolite using thermal treatment (550°C, 5h) and this material has excellent performance as an antibacterial agent after silver ions loading.

Matched MeSH terms: Escherichia coli/growth & development*

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