Search results for: isolated footings

Authors: Firoz Alam Faroque, Ankur Neog

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Earthquake causes significant loss of lives and severe damage to infrastructure. Base isolator is one of the most suitable solutions to make a building earthquake resistant. Base isolation consists of installing an isolator along with the steel plates covered with pads of strong material like steel, rubber, etc. In our study, we have used lead rubber bearing (LRB). The basic idea of seismic isolation is based on the reduction of the earthquake-induced inertia forces by shifting the fundamental period of the structure out of dangerous resonance range, and concentration of the deformation and energy dissipation demands at the isolation and energy dissipation systems, which are designed for this purpose. In this paper, RC frame buildings have been modeled and analyzed by response spectrum method using ETABS software. The LRB used in the model is designed as per uniform building code (UBC) 97. It is found that time period for the base isolated structures are higher than that of the fixed base structure and the value of base shear significantly reduces in the case of base-isolated buildings. It has also been found that buildings with vertical irregularities give better performance as compared to building with plan irregularities using base isolators.

Keywords: base isolation, base shear, irregularities in buildings, lead rubber bearing (LRB)

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Authors: Ilaisa Kacivakanadina, Samson Viulu, Brad Carte, Katy Soapi

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Two bioactive compounds namely Vulgamycin (also known as enterocin A) and 5-deoxyenterocin were purified from a marine bacterial strain 1903. Strain 1903 was isolated from marine sediments collected from the Solomon Islands. Morphological features of strain 1903 showed that it belongs to the genus Streptomyces. The two secondary metabolites were extracted using EtOAc and purified by chromatographic methods using EtOAc and hexane solvents. Mass spectrum and NMR data of pure compounds were used to elucidate the chemical structures. In this study, results showed that both compounds were strongly active against Wild Type Staphylococcus aureus (WTSA) (MIC < 1 µg/mL) and in Brine shrimp assays (BSA) (MIC < 1 µg/mL). 5-deoxyenterocin was also active against Rifamycin resistant Staphylococcus aureus (RRSA) (MIC, 250 µg/mL) while vulgamycin showed bioactivity against Methicillin resistant Staphylococcus aureus (MRSA) (MIC 250 µg/mL). To the best of our knowledge, this is the first study that showed the bio-activity of 5-deoxyenterocin. This is also the first time that Vulgamycin has been reported to be active in a BSA. There has not been any mechanism of action studies for these two compounds against pathogens. This warrants further studies on their mechanism of action against microbial pathogens.

Keywords: 5-deoxyenterocin, bioactivity, brine shrimp assay (BSA), vulgamycin

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Authors: Elisângela de Albuquerque Sobreira Borovoski, Ricardo Willian Borovoski

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Microbiological infections acquired by animals pose a risk to public health. In public health, monitoring the health of primates is linked to the risk of transmission of zoonoses through scratches, bites and contact with biological samples. The project was approved by the Ethics Committee on the Use of Animals Protocol No. 170/2019. It was authorized by ICMBio Protocol No. 52117-1. The study was carried out in the period 2019-2022 in the municipality of Anápolis. Iron and galvanized wire traps were used and the animals were anesthetized with 4.4mg/kg zolethyl intramuscularly and saliva was collected through swabs. Fifty-three capuchin monkeys were captured from the Onofre Quinan Environmental Park in Anápolis-Goiás for health monitoring purposes. In the laboratory, the samples were deposited on the agar surface and seeded by exhaustion to obtain isolated colonies. These colonies were analyzed according to morphocolonial characteristics. Morphometric characterization and biochemical tests for bacterial identification were performed. A total of 861 bacterial samples were isolated, nine of which were strict anaerobic bacteria of the genus Peptostreptococcus. Previous and constant knowledge of the prevalence of pathogenic agents in biological samples is essential to be prepared to act in pandemic situations.

Keywords: Brazil, microbiology, monkeys, public health

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Authors: Akwu A. Nneka, Naidoo, Yougasphree

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A significantly high number of microbes exist in higher plants; these microbes include bacteria, fungi, and actinomycetes. There are reports on the benefits of endophytic fungi and their products of metabolism to the host plant and man, consequently, it is expedient to explore the changes that could arise as a result of manipulating their growth media. Grewia lasiocarpa E. Mey. ex Harv. (Malvaceae) is an indigenous Southern African plant, that belongs to a genus with known medicinal properties. Three media were used to culture the endophytic fungi viz., Potato Dextrose Agar (PDA), Malt Extract Agar (MEA), and Bacteriological Agar (BA) were used singly, and supplemented with three dilutions of the leaves and stem bark extracts. The manipulated growth media composition had a significant effect on the diversity of the isolated fungal populations. Several endophytic fungi were isolated; their distribution and diversity revealed a significant relatedness with the manipulated media. The media supplemented with the plant extracts was observed to give a significant increase in the growth rate and yield of the endophytes. To the best of our knowledge, this is the first study describing the endophytic fungi present in the leaves and stem bark of G. lasiocarpa E. Mey. ex Harv.

Keywords: Grewia lasiocarpa, plant-based extracts, endophytic fungi, Malvaceae

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Authors: Sangram Shamrao Patil, Hara Mohan Jena

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Phthalates are among the most common organic pollutant since they have become widespread in the environment and found in sediments, natural waters, soils, plants, landfill leachates, biota including human tissue and aquatic organisms. Diethyl phthalate (DEP) is a low molecular weight phthalate which has wide applications as plasticizer and become a major cause of environmental pollution. Environmental protection agency (EPA) listed DEP as priority pollutant because of its toxicity and they recommended human health ambient water quality criterion for diethyl phthalate (DEP) as 4 mg/l. Therefore, wastes containing phthalates require proper treatment before being discharged into the environment. Biodegradation is attractive and efficient treatment method as it is cost effective and produces non-toxic end products. In the present study, a DEP degrading aerobic bacterium was isolated from soil contaminated with plastic industry wastewater. Morphological and biochemical characteristics of isolate were performed. 16S rRNA sequencing and phylogenetic analysis of isolate was carried out and it was identified as Empedobacter brevis. Isolate has been found to tolerate up to 1650 mg/l of DEP. This study will be significant for exploring an application of microbes for remediation of phthalates and development of a suitable bioreactor.

Keywords: diethyl phthalate, plasticizer, pollutant, biodegradation

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Authors: Sahira Nsayef Muslim, Israa M. S. Al-Kadmy, Nadheema Hammood Hussein, Alaa Naseer Mohammed Ali, Buthainah Mohammed Taha, Rayim Sabah Abbood, Sarah Naji Aziz

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A novel strain of Bacillus licheniformis isolated from spoilt cucumber and pepper samples have the ability to produce the chitosanase enzyme when grown on chitosan substrate. Chitosanase was purified to homogeneity with a recovery yield of 35.71% and 5.5 fold of purification by using ammonium sulfate at 45% saturation followed by ion exchange chromatography on DEAE-cellulose column and gel filtration chromatography on Sephadex G-100 column. The purified chitosanase inhibited the biofilm formation ability for all Gram-negative and Gram-positive biofilm-forming bacteria (biofilm producers) after using Congo Red agar and Microtiter plates methods. Highly antibiofilm of chitosanase recorded against Pseudomonas aeruginosa followed by Klebsiella pneumoniae with reduction of biofilm formation ratio to 22 and 29%, respectively compared with (100)% of control. Thus, chitosanase has promising benefit as antibiofilm agent against biofilm forming pathogenic bacteria and has promising application as alternative antibiofilm agents to combat the growing number of multidrug-resistant pathogen-associated infections, especially in situation where biofilms are involved.

Keywords: chitosanase, Bacillus licheniformis, vegetables, biofilm

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Authors: Vishvas N. Patel, Mehul Chorawala

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Currently, according to the authors best knowledge there are less effective therapeutic agents to limit intestinal mucosa damage associated with inflammatory bowel disease (IBD). Clinical studies have shown beneficial effects of several probiotics in patients of IBD. Probiotics are live organisms; confer a health benefit to the host by modulating immunoinflammation and oxidative stress. Although probiotics in murine and human improve disease severity, very little is known about the specific contribution of cell wall contents of probiotics in IBD. Herein, we investigated the ameliorative potential of cell wall contents of Lactobacillus casei (LC) in lipopolysaccharide (LPS)-induced murine colitis. Methods: Colitis was induced in LPS-sensitized rats by intracolonic instillation of LPS (50 µg/rat) for consecutive 14 days. Concurrently, cell wall contents isolated from 103, 106 and 109 CFU of LC was given subcutaneously to each rat for 21 days, considering sulfasalazine (100 mg/kg, p.o.) as standard. The severity of colitis was assessed by body weight loss, food intake, stool consistency, rectal bleeding, colon weight/length, spleen weight and histological analysis. Colonic inflammatory markers (myeloperoxidase (MPO) activity, C-reactive protein and proinflammatory cytokines) and oxidative stress markers (malondialdehyde, reduced glutathione and nitric oxide) were also assayed. Results: Cell wall contents of isolated from 106 and 109 CFU of LC significantly improved the severity of colitis by reducing body weight loss and diarrhea & bleeding incidence, improving food intake, colon weight/length, spleen weight and microscopic damage to the colonic mucosa. The treatment also reduced levels of inflammatory and oxidative stress markers and boosted antioxidant molecule. However, cell wall contents of isolated from 103 were ineffective. Conclusion: In conclusion, cell wall contents of LC attenuate LPS-induced colitis by modulating immuno-inflammation and oxidative stress.

Keywords: probiotics, Lactobacillus casei, immuno-inflammation, oxidative stress, lipopolysaccharide, colitis

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Authors: Ikram Mohamed Eltayeb Elsiddig, Abdel Khalig Muddather, Hiba Abdel Rahman Ali, Saad Mohamed Hussein Ayoub

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Anogeissusleiocarpus (Combretaceae) is well known for its medicinal uses in African traditional medicine, for treating many human diseases mainly skin diseases and infections.Mycetoma disease is a fungal and/ or bacterial skin infection, mainly cause by Madurella mycetomatis fungus.This study was carried out in vitro to investigate the antifungal activity of Anogeissusleiocarpus leaf extracts against the isolated pathogenicMadurellamycetomatis, by using the NCCLS modified method compared to Ketoconazole standard drug and MTT assay. The bioactive fraction was subjected to chemical analysis implementing different chromatographic analytical methods (TLC, HPLC, and LC-MS/MS). The results showed significance antifungal activity of A. leiocarpus leaf extractsagainst the isolated pathogenicM. mycetomatis, compared to negative and positive controls. The chloroform fraction showed the highest antifungal activity.The chromatographic analysis of the chloroform fraction with the highest activity showed the presence of important bioactive compounds such as ellagic and flavellagic acids derivatives, flavonoids and stilbenoid, which are well known for their antifungal activity.

Keywords: Anogeissus leiocarpus, crude extracts and fractions of Anogeissus leiocarpus, in vitrosusceptibility of Madurella mycetomatis, Madurella mycetomatis

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Authors: A. Nkohla, U. Nwodo, L. V. Mabinya, A. I. Okoh

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A potential source for low-cost production of value added products is the utilization of lignocellulosic materials. However, the huddle needing breaching would be the dismantlement of the complex lignocellulosic structure as to free sugar base therein. the current lignocellosic material treatment process is expensive and not eco-friendly hence, the advocacy for enzyme based technique which is both cheap and eco-friendly is highly imperative. Consequently, this study aimed at the screening of cellulose and xylan degrading bacterial strain isolated from decaying sawdust samples. This isolate showed high activity for cellulase and xylanase when grown on carboxymethyl cellulose and birtchwood xylan as the sole carbon source respectively. The 16S rDNA nucleotide sequence of the isolate showed 98% similarity with that of Chryseobacterium taichungense thus, it was identified as a Chryseobacterium sp. Optimum culture conditions for cellulase and xylanase production were medium pH 6, incubation temperature of 25 °C at 50 rpm and medium pH 6, incubation temperature of 25 °C at 150 rpm respectively. The high enzyme activity obtained from this bacterial strain portends it as a good candidate for industrial use in the degradation of complex biomass for value added products.

Keywords: lignocellulosic material, chryseobacterium sp., submerged fermentation, cellulase, xylanase

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Authors: Ali A. Al-Jazaairry, Tahsin T. Sabbagh

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One of the important concerns within the field of geotechnical engineering is the presence of cavities in soils. This present work is an attempt to understand the behaviour of strip footing subjected to inclined load and constructed on cavitied soil. The failure mechanism of strip footing located above such soils was studied analytically. The capability of analytical model to correctly expect the system behaviour is assessed by carrying out verification analysis on available studies. The study was prepared by finite element software (PLAXIS) in which an elastic-perfectly plastic soil model was used. It was indicated, from the results of the study, that the load carrying capacity of foundation constructed on cavity can be analysed well using such analysis. The research covered many foundation cases, and in each foundation case, there occurs a critical depth under which the presence of cavities has shown minimum impact on the foundation performance. When cavities are found above this critical depth, the load carrying capacity of the foundation differs with many influences, such as the location and size of the cavity and footing depth. Figures involving the load carrying capacity with the affecting factors studied are presented. These figures offer information beneficial for the design of strip footings rested on underground cavities. Moreover, the results might be used to design a shallow foundation constructed on cavitied soil, whereas the obtained failure mechanisms may be employed to improve numerical solutions for this kind of problems.

Keywords: axial load, cavity, inclined load, strip footing

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Authors: Mohamed M. Alsull

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In the present study, two microalgae species “Nannochloropsis sp. and Tetraselmis sp.” isolated from Penang National Park coastal waters, Malaysia; were cultivated under combined various laboratory conditions “salinity, light, nitrogen limitation and starvation”. Growth rate, dry weight, chlorophyll a content, total lipid and protein contents, were estimated at mid exponential growth phase. Both Nannochloropsis sp. and Tetraselmis sp. showed remarkable decrease in growth rate, chlorophyll a content and protein content companied with increase in lipid content under nitrogen limitation and starvation conditions. Maintaining Nannochloropsis sp. under salinity 15‰ caused only significant decrease in total protein content; while Tetraselmis sp. grown at the same salinity caused decrease in the growth rate, chlorophyll a, dry weight and total protein content only when nitrogen was available.

Keywords: biochemical composition, light, microalgae, nitrogen limitation, salinity

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Authors: Maha Soltan, Amal Z. Hassan, Howaida I. Abd-Alla, Atef G. Hanna

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Two naturally known cardenolides; acovenoside A and acobioside A were isolated from the Egyptian cultivar; Acokanthera spectabilis leaves. It is an ornamental and poisonous plant that has been traditionally claimed for their medicinal properties against infectious microbes, killing worms and curing some inflammations at little amounts. We examined the growth inhibition effects of both cardenolides against four types of human cancer cell lines using Sulphorhodamine B assay. In addition, the clonogenic assay was also performed for testing the growth inhibiting power of the isolated compounds. An in vitro mechanistic investigation was further accomplished against hepatocellular carcinoma HepG2 cell line. Microscopic examination, colorimetric ELISA and flow cytometry techniques were our tools of proving at least part of the anticancer pathway of the tested compounds. Both compounds were able to inhibit the growth of 4 human cancer cell lines at less than 100 nM. In addition, they were able to activate the executioner Caspase-3 and apoptosis was then induced as a consequence of cell growth arrest at G2/M. An attention must be payed to those bioactive agents particularly when giving their activity against cancer cells at considerable small values while presenting safe therapeutic margins as indicated by literature.

Keywords: anticancer, cardenolides, Caspase-3, apoptosis

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Authors: Sohan Sengupta, Arnab Pramanik, Abhrajyoti Ghosh, Maitree Bhattacharyya

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Newly emerged phyto-pathogens and multi drug resistance have been threating the world for last few decades. Actinomycetes, the most endowed group of microorganisms isolated from unexplored regions of the world may be the ultimate solution to these problems. Thus the aim of this study was to isolate several bioactive actinomycetes strains capable of producing antimicrobial secondary metabolite from Sundarbans, the only mangrove tiger land of the world. Fifty four actinomycetes were isolated and analyzed for antimicrobial activity against fifteen test organisms including three phytopathogens. Nine morphologically distinct and biologically active isolates were subjected to polyphasic identification study. 16s rDNA sequencing indicated eight isolates to reveal maximum similarity to the genus streptomyces, whereas one isolate presented only 93.57% similarity with Streptomyces albogriseolus NRRL B-1305T. Seventy-one carbon sources and twenty-three chemical sources utilization assay revealed their metabolic relatedness. Among these nine isolates three specific strains were found to have notably higher degree of antimicrobial potential effective in a broader range including phyto-pathogenic fungus. PCR base whole genome screen for PKS and NRPS genes, confirmed the occurrence of bio-synthetic gene cluster in some of the isolates for novel antibiotic production. Finally the strain SMS_SU21, which showed antimicrobial activity with MIC value of 0.05 mg ml-1and antioxidant activity with IC50 value of 0.242±0.33 mg ml-1 was detected to be the most potential one. True prospective of this strain was evaluated utilizing GC-MS and the bioactive compound responsible for antimicrobial activity was purified and characterized. Rare bioactive actinomycetes were isolated from unexplored heritage site. Diversity of the biosynthetic gene cluster for antimicrobial compound production has also been evaluated. Antimicrobial compound SU21-C has been identified and purified which is active against a broad range of pathogens.

Keywords: actinomycetes, sundarbans, antimicrobial, pks nrps, phyto-pathogens, GC-MS

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Authors: Mulata Haile Nega, Derebew Fikadu Berhe, Vera Ribeiro Marques

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There is no epidemiologic data on this gene polymorphism in several countries. Therefore, this study aimed to assess the genotype and allele frequencies of the gene variant in three countries. This study involved healthy individuals from Colombia, Mozambique, and Portugal. Genomic DNA was isolated from blood samples using the Qiamp DNA Extraction Kit (Qiagen). The isolated DNA was genotyped using Polymerase Chain Reaction (PCR) - Restriction Fragment Length Polymorphism. Microstat and GraphPad quick cal software were used for the Chi-square test and evaluation of Hardy-Weinberg equilibrium, respectively. A total of 181 individuals’ blood sample was analyzed. Overall, TT (74.0%) genotype was the highest, and CC (7.8%) was the lowest. Country wise genotypic frequencies were Colombia 47(70.2%) TT, 12(17.9%) TC and 8(11.9%) CC; Mozambique 47(88.7%) TT, 5(9.4%) TC, and 1(1.9%) CC; and Portugal 40(65.6%) TT, 16(26.2%) TC, and 5(8.2%) CC. The reference (T) allele was highest among Mozambicans (93.4%) compared to Colombians (79.1%) and Portuguese (78.7%). Mozambicans showed statistically significant genotypic and allelic frequency differences compared to Colombians (p<0.01) and Portuguese (p <0.01). Overall and country-wise, the CC genotype was less frequent and relatively high for Colombians and Portuguese populations. This finding may imply statins risk-benefit variability associated with CC genotype among these populations that needs further understanding.

Keywords: c.521T>C, polymorphism, SLCO1B1, SNP, statins

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Authors: Qiang Xu, Xingxin Wu, Wenjie Guo, Xingqi Wang, Yang Sun, Renxiang Tan

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The phosphatase SHP-2 is known to exert regulatory activities on cytokine receptor signaling and the dysregulation of SHP-2 has been implicated in the pathogenesis of a variety of diseases. Here we report several small molecule regulators of SHP-2 for the treatment of T cell-mediated diseases. The new cyclodepsipeptide trichomides A, isolated from the fermentation products of Trichothecium roseum, increased the phosphorylation of SHP-2 in activated T cells, and ameliorated contact dermatitis in mice. The trichomides A’s effects were significantly reversed by using the SHP-2-specific inhibitor PHPS1 or T cell-conditional SHP-2 knockout mice. Another compound is a cerebroside Fusaruside isolated from the endophytic fungus Fusarium sp. IFB-121. Fusaruside also triggered the tyrosine phosphorylation of SHP-2, which provided a possible mean of selectively targeting STAT1 for the treatment of Th1 cell-mediated inflammation and led to the discovery of the non-phosphatase-like function of SHP-2. Namely, the Fusaruside-activated pY-SHP-2 selectively sequestrated the cytosolic STAT1 to prevent its recruitment to IFN-R, which contributed to the improvement of experimental colitis in mice. Blocking the pY-SHP-2-STAT1 interaction, with SHP-2 inhibitor NSC-87877 or using T cells from conditional SHP-2 knockout mice, reversed the effects of fusaruside. Furthermore, the fusaruside’s effect is independent of the phosphatase activity of SHP-2, demonstrating a novel role for SHP-2 in regulating STAT1 signaling and Th1-type immune responses.

Keywords: SHP-2, small molecules, T cell, T cell-mediated diseases

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Authors: Jma Hannan, Prawej Ansari, Yasser H. A. Abdel-Wahab, Peter R. Flatt

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Spirulina platensis (SP, Blue-green algae) have been accepted as a supplement for the treatment of pre and post-diabetes. The present study investigated the effects of butanol fraction from ethanol extract of S. platensis on insulin release from BRIN BD11 cells, isolated mouse islets, and perfused rat pancreas, as well as glucose homeostasis in type 2 diabetic rats and their molecular pathways. In a dose-dependent manner, S. platensis increased insulin release from mouse islets and pancreatic β-cells. The extract also elevated insulin release in perfused rat pancreas. Glucose, isobutylmethylxanthine, tolbutamide, and a depolarizing concentration of KCl significantly potentiated insulin release from BRIN BD11 cells. The effect of diazoxide and verapamil, as well as the absence of extracellular Ca2+ showed a reduction in insulin secretion. When administered orally together with glucose (2.5g/kg bw), S. platensis extract improved fasting and postprandial blood glucose in type 2 diabetes. These data suggest that the anti-diabetic activity of S. platensis is partly mediated by insulin secretion via the KATP channel-dependent pathway/the intracellular cAMP pathway.

Keywords: Insulin, glucose, S. platensis, type 2 diabetes, medicinal plants

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Authors: Carol N. Flores-Fernández, Guadalupe Espilco, Cynthia Esquerre, Amparo I. Zavaleta

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Saline environments represent a valuable source of enzymes with novel properties and particular features for application in food, pharmaceutical and chemical industry. This study focuses on the isolation and screening of hydrolase-producing bacteria from Chilca salterns and the evaluation of their biotechnological potential. Soil samples were collected from Chilca salterns in Peru. For the isolation, medium containing 0.2 % of yeast extract, 5 % of NaCl and 10 % of the soil sample was used. After 72 h of incubation at 37 °C, serial dilutions were made up to 10−12 dilutions, spread on agar plates with 0.5 % of yeast extract and 5 % of NaCl, and incubated at 37 °C for 48 h. Screening of hydrolase-producing bacteria was carried out for cellulases, amylases, lipases, DNase, and proteases on specific media. Moreover, protease-producing bacteria were tested using protein extracted from the following legumes as substrate: Glycine max, Lupinus mutabilis, Pisum sativum, Erythrina edulis, Cicer arietinum, Phaseolus vulgaris and Vicia faba. A total of 16 strains were isolated from soil samples. On the screening media; 75, 44, 81 and 50 % were cellulase, amylase, DNase and protease producers, respectively. Also, 19 % of the isolates produced all the hydrolytic enzymes above mentioned. Lipase producers were not found. The 37 % and 12 % of the strains grew at 20 % and 30 % of salt concentration, respectively. In addition, 75 % of the strains grew at pH range between 5 and 10. From the total of protease-producing bacteria, 100 % hydrolyzed Glycine max, Lupinus mutabilis, and Pisum sativum protein, while 87 % hydrolyzed Erythrina edulis and Cicer arietinum protein. Finally, 75 % and 50 % of the strains hydrolyzed Phaseolus vulgaris and Vicia faba protein, respectively. Hydrolase-producing bacteria isolated from Chilca salterns in Peru grew at high salt concentrations and wide range of pH. In addition, protease-producing bacteria hydrolyzed protein from different sources such as leguminous. These enzymes have great biotechnological potential and could be used for different industrial processes and applications.

Keywords: bacteria, extracellular, hydrolases, Peru, salterns

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Authors: Jin-Hyeong Park, Hong-Seok Kim, Jin-Hyeok Yim, Young-Ji Kim, Dong-Hyeon Kim, Jung-Whan Chon, Kun-Ho Seo

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Salmonella contamination in chicken samples can cause major health problems in humans. However, not only the effects of antibiotic treatment during growth but also the impacts of poultry slaughter line on the prevalence of Salmonella in final chicken meat sold to consumers are unknown. In this study, we compared the isolation rates and antimicrobial resistance of Salmonella between antibiotic-free, conventional, conventional Korean native retail chicken meat samples and clonal divergence of Salmonella isolates by multilocus sequence typing. In addition, the distribution of extended-spectrum β-lactamase (ESBL) genes in ESBL-producing Salmonella isolates was analyzed. A total of 72 retail chicken meat samples (n = 24 antibiotic-free broiler [AFB] chickens, n = 24 conventional broiler [CB] chickens, and n = 24 conventional Korean native [CK] chickens) were collected from local retail markets in Seoul, South Korea. The isolation rates of Salmonella were 66.6% in AFB chickens, 45.8% in CB chickens, and 25% in CK chickens. By analyzing the minimum inhibitory concentrations of β -lactam antibiotics with the disc-diffusion test, we found that 81.2% of Salmonella isolates from AFB chickens, 63.6% of isolates from CB chickens, and 50% of isolates from CK chickens were ESBL producers; all ESBL-positive isolates had the CTX-M-15 genotype. Interestingly, all ESBL-producing Salmonella were revealed as ST16 by multilocus sequence typing. In addition, all CTX-M-15-positive isolates had the genetic platform of blaCTX-M gene (IS26-ISEcp1-blaCTX-M-15-IS903), to the best of our knowledge, this is the first report in Salmonella around the world. The Salmonella ST33 strain (S. Hadar) isolated in this study has never been reported in South Korea. In conclusion, our findings showed that antibiotic-free retail chicken meat products were also largely contaminated with ESBL-producing Salmonella and that their ESBL genes and genetic platforms were the same as those isolated from conventional retail chicken meat products.

Keywords: antibiotic-free poultry, conventional poultry, multilocus sequence typing, extended-spectrum β-lactamase, antimicrobial resistance

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Authors: T. Warinpramote, J. Sanguanjeen, P. Pholphakwaen, S. Kittisorayut, J. Kerdtubtim, S. Palachote, M. Taweechotipatr

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Cholesterol is a type of lipid molecule which is the significant risk factor for coronary heart disease. Currently, there are many cholesterol-lowering alternative treatments especially bile salt hydrolase positive lactobacilli. BSH can cleave the peptide linkage of bile salt, which results in removal of the amino acid group from the steroid core and increases production of the new bile acid by using more cholesterol. The purpose of this study was to isolate, and screen probiotic characteristics of lactobacilli from fermented Thai foods and further investigated for their comparative BSH activity. The result showed that 2 of 81 Lactobacillus strains, JPK2-2 and JPK3-2, isolated from Brassica juncea (L.) had significantly exhibited high BSH activity. In addition, these Lactobacillus strains showed their results that the ability to tolerate acid and bile salt. Furthermore, the using of 16S rDNA sequencing for definitive microbial identifications showed that these 2 strains belong to Lactobacillus plantarum. In the future, the L. plantarum with BSH activity strains JPK2-2 and JPK3-2 may be the candidate probiotics for application in functional foods and dairy products to improve in the patient with cardiovascular diseases.

Keywords: Lactobacillus plantarum, probiotics, bile salt hydrolase, cholesterol-lowering, fermented Thai food

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Authors: S. J. Sirima, C. Thirawan, R.Puntharikorn, K. Ungsumalin, J. Kaemwich

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Acinetobacter spp. is a gram negative bacterium causing the high incidence of multi-drug resistance in patients admitted to an intensive care unit. A hundred isolates of Imipenem-resistant Acinetobacter spp. isolated from patients admitted at tertiary health care center, Northeastern region, Ubon Ratchathani, Thailand, were subjected to modified Hodge test and combined disc test in order to evaluate the production of carbapenemases. The results revealed that about 35% of isolates were found to be carbapenemases producers. In addition, multiplex polymerase chain reactions were performed to detect blaOXA-like genes. It showed that 92% of isolates possess blaOXA-51-like and blaOXA-23-like genes. However, blaOXA-58-like gene was detected in only 8 isolates. No detection of blaOXA-24-like gene was observed in all isolates. In conclusion, an ability to produce carbepenemases would be an important mechanism of multi-drug resistance among clinical isolates of Acinetobacter spp. at tertiary health care center, Northeastern region, Ubon Ratchathani, Thailand. Furthermore, it was likely that the class D carbapenemases genes, blaOXA-51-like and blaOXA-23-like, might contribute to imipenem-resistance exhibiting among isolates.

Keywords: Acinetobacter spp., blaOXA-like genes, carbapenemases, tertiary health care center

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Authors: Ashraf Y. Z. Khalifa, Mohammed A. Almalki

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Medicago sativa (Alfalfa) is an important forage crop legume worldwide including Saudia Arabia due to its high nutritive value. Soil bacteria exist in root or root-nodules of Medicago sativa in either symbiotic relationships or in associations. The aim of the present study was to isolate and characterize endophytic bacteria that live in association with non-nodulated roots of Medicago sativa growing in Al-Ahsaa region, Saudia Arabia. Several bacterial strains were isolated from sterilized roots of Medicago sativa. Strains were characterized using 16S rRNA gene sequences, phylogenetic relationships analysis, morphological and biochemical characteristics. The strains utilized 50% (10 out of 20) of the different chemical substrates contained in the API20E strip. In general, many strains had the ability to ferment/oxidise all the carbohydrate tested except for rhamnose and the polyol carbohydrate, inositol. Comparative sequence analysis of the 16S rDNA gene indicated that the strains were closely related to the genus Bacillus. Furthermore, the growth parameters of Vigna sinensis were enhanced upon single-inoculation of the isolated strains, compared to the uninoculated control plants. The results highlighted that the root-nodules of Medicago sativa harbor non-nodulating bacterial strains that could have significant agricultural applications.

Keywords: Medicago sativa, endophytic bacteria, Pisum sativum, Vigna sinensis

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Authors: Walaa Al-Maghraby

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Endophytes are organisms that colonize internal plant tissues without causing apparent harm to their host. Almost all groups of microorganisms have been found in endophytic association with plants may be fungi. They stimulate the production of secondary metabolites with a diverse range of biological activities. Leptadenia pyrotechnica is a more or less leafless, erect shrub with straight stems which is highly distributed in Saudi Arabia. Four endophytes fungi were isolated from Leptadenia pyrotechnica and identified using 18S ribosomal RNA sequences, which revealed four fungi genuses, namely Aspergillus terreus; Aspergillus welwitschiae; Aspergillus fumigatus and Aspergillus flavus. In this present study, four endophytic fungi from Leptadenia pyrotechnica were used for obtaining crude aqueous and ethyl acetate extracts for antimicrobial screening against 6 human pathogens, the antibacterial tests presented satisfactory results, where the pathogenic bacteria were inhibited by the four extracts tested, except for Escherichia coli that was inhibited by all extracts except ethyl acetate extract of Aspergillus terreus. Analysis of variance showed that the extract produced by endophyte Leptadenia pyrotechnica was the most effective against all bacteria, either gram-negative or positive. However, the extract was not efficient against pathogenic fungi. Therefore, this study indicates that endophytes from medicinal plant Leptadenia pyrotechnica could be potential sources of antibacterial substances.

Keywords: antimicrobial activity, Aspergillus sp, endophytes, Leptadenia pyrotechnica

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Authors: Kaustuvmani Patowary, Suresh Deka

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Oil pollution accidents, nowadays, have become a common phenomenon and have caused ecological and social disasters. Microorganisms with high oil-degrading performance are essential for bioremediation of petroleum hydrocarbon. In this investigation, an effective biosurfactant producer and hydrocarbon degrading bacterial strain, Pseudomonas sp.PG1 (identified by 16s rDNA sequencing) was isolated from hydrocarbon contaminated garage soil of Pathsala, Assam, India, using crude oil enrichment technique. The growth parameters such as pH and temperature were optimized for the strain and upto 81.8% degradation of total petroleum hydrocarbon (TPH) has been achieved after 5 weeks when grown in mineral salt media (MSM) containing 2% (w/v) crude oil as the carbon source. The biosurfactant production during the course of hydrocarbon degradation was monitored by surface tension measurement and emulsification activity. The produced biosurfactant had the ability to decrease the surface tension of MSM from 72 mN/m to 29.6 mN/m, with the critical micelle concentration (CMC)of 56 mg/L. The biosurfactant exhibited 100% emulsification activity on crude oil. FTIR spectroscopy and LCMS-MS analysis of the purified biosurfactant revealed that the biosurfactant is Rhamnolipidic in nature with several rhamnolipid congeners. Gas Chromatography-Mass spectroscopy (GC-MS) analysis clearly demonstrated that the strain PG1 efficiently degrade different hydrocarbon fractions of the crude oil. The study suggeststhat application of the biosurfactant producing strain PG1 as an appropriate candidate for bioremediation of crude oil contaminants.

Keywords: petroleum hydrocarbon, hydrocarbon contamination, bioremediation, biosurfactant, rhamnolipid

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Authors: Rafia S. Al-lamki, Jun Wang, Simon Pacey, Jordan Pober, John R. Bradley

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Tumor necrosis factor alpha (TNF), signaling through TNFR2, may act an autocrine growth factor for renal tubular epithelial cells. Clear cell renal carcinomas (ccRCC) contain cancer stem cells (CSCs) that give rise to progeny which form the bulk of the tumor. CSCs are rarely in cell cycle and, as non-proliferating cells, resist most chemotherapeutic agents. Thus, recurrence after chemotherapy may result from the survival of CSCs. Therapeutic targeting of both CSCs and the more differentiated bulk tumor populations may provide a more effective strategy for treatment of RCC. In this study, we hypothesized that TNFR2 signaling will induce CSCs in ccRCC to enter cell cycle so that treatment with ligands that engage TNFR2 will render CSCs susceptible to chemotherapy. To test this hypothesis, we have utilized wild-type TNF (wtTNF) or specific muteins selective for TNFR1 (R1TNF) or TNFR2 (R2TNF) to treat either short-term organ cultures of ccRCC and adjacent normal kidney (NK) tissue or cultures of CD133+ cells isolated from ccRCC and adjacent NK, hereafter referred to as stem cell-like cells (SCLCs). The effect of cyclophosphamide (CP), currently an effective anticancer agent, was tested on CD133+SCLCs from ccRCC and NK before and after R2TNF treatment. Responses to TNF were assessed by flow cytometry (FACS), immunofluorescence, and quantitative real-time PCR, TUNEL, and cell viability assays. Cytotoxic effect of CP was analyzed by Annexin V and propidium iodide staining with FACS. In addition, we assessed the effect of TNF on isolated SCLCs differentiation using a three-dimensional (3D) culture system. Clinical samples of ccRCC contain a greater number SCLCs compared to NK and the number of SCSC increases with higher tumor grade. Isolated SCLCs show expression of stemness markers (oct4, Nanog, Sox2, Lin28) but not differentiation markers (cytokeratin, CD31, CD45, and EpCAM). In ccRCC organ cultures, wtTNF and R2TNF increase CD133 and TNFR2 expression and promote cell cycle entry whereas wtTNF and R1TNF increase TNFR1 expression and promote cell death of SCLCs. Similar findings are observed in SCLCs isolated from NK but the effect was greater in SCLCs isolated from ccRCC. Application of CP distinctly triggered apoptotic and necrotic cell death in SLCSs pre-treatment with R2TNF as compared to CP treatment alone, with SCLCs from ccRCC more sensitive to CP compared to SLCS from NK. Furthermore, TNF promotes differentiation of SCLCs to an epithelial phenotype in 3D cultures, confirmed by cytokeratin expression and loss of stemness markers Nanog and Sox2. The differentiated cells show positive expression of TNF and TNFR2. These findings provide evidence that selective engagement of TNFR2 drive CSCs to cell proliferation/differentiation, and targeting of cycling cells with TNFR2 agonist in combination with anti-cancer agents may be a potential therapy for RCC.

Keywords: cancer stem cells, ccRCC, cell cycle, cell death, TNF, TNFR1, TNFR2, CD133

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Authors: Karol Rodriguez-Peña, Martha L. Macias-Rubalcava, Leticia Rocha-Zavaleta, Sergio Sanchez

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A bioassay-guided study for anti-cancer compounds from endophytes of the Mexican medicinal plant Amphipteryygium adstringens resulted in the isolation of a streptomycete capable of producing a group of compounds with high cytotoxic activity. Microorganisms from surface sterilized samples of various sections of the plant were isolated and all the actinomycetes found were evaluated for their potential to produce compounds with cytotoxic activity against cancer cell lines MCF7 (breast cancer) and HeLa (cervical cancer) as well as the non-tumoural cell line HaCaT (keratinocyte). The most active microorganism was picked for further evaluation. The identification of the microorganism was carried out by 16S rDNA gene sequencing, finding the closest proximity to Streptomyces scabrisporus, but with the additional characteristic that the strain isolated in this study was capable of producing colorful compounds never described for this species. Crude extracts of dichloromethane and ethyl acetate showed IC50 values of 0.29 and 0.96 μg/mL for MCF7, 0.51 and 1.98 μg/mL for HeLa and 0.96 and 2.7 μg/mL for HaCaT. Scaling the fermentation to 10 L in a bioreactor generated 1 g of total crude extract, which was fractionated by silica gel open column to yield 14 fractions. Nine of the fractions showed cytotoxic activity. Fraction 4 was chosen for subsequent purification because of its high activity against cancerous cell lines, lower activity against keratinocytes. HPLC-UV-MS/ESI was used for the evaluation of this fraction, finding at least 10 different compounds with high values of m/z (≈588). Purification of the compounds was carried out by preparative thin-layer chromatography. The prevalent compound was Steffimycin B, a molecule known for its antibiotic and cytotoxic activities and also for its low solubility in aqueous solutions. Along with steffimycin B, another five compounds belonging to the steffimycin family were isolated and at this moment their structures are being elucidated, some of which display better solubility in water: an attractive property for the pharmaceutical industry. As a conclusion to this study, the isolation of endophytes resulted in the discovery of a strain capable of producing compounds with high cytotoxic activity that need to be studied for their possible utilization.

Keywords: amphipterygium adstringens, cytotoxicity, streptomyces scabrisporus, steffimycin

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Authors: Pampita Chakraborty, Sukumar Mukherjee

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People with diabetes mellitus are more susceptible to developing infections, as high blood sugar levels can weaken the patient's immune system defences. People with diabetes are more adversely affected when they get an infection than someone without the disease, because you have weakened immune defences in diabetes. People who have minimally elevated blood sugar levels experience worse outcomes with infections. Diabetic patients in hospitals do not necessarily have a higher mortality rate due to infections, but they do face longer hospitalisation and recovery times. A study was done in a tertiary care unit in eastern India. Patients with type 2 diabetes mellitus infection were recruited in the study. A total of 520 cases of Type 2 Diabetes Mellitus were recorded out of which 200 infectious cases was included in the study. All subjects underwent detailed history & clinical examination. Microbiological samples were collected from respective site of the infection for microbial culture and antibiotic sensitivity test. Out of the 200 infectious cases urinary tract infection(UTI) was found in majority of the cases followed by diabetic foot ulcer (DFU), respiratory tract infection(RTI) and sepsis. It was observed that Escherichia coli was the most commonest pathogen isolated from UTI cases and Staphylococcus aureus was predominant in foot ulcers followed by other organisms. Klebsiella pneumonia was the major organism isolated from RTI and Enterobacter aerogenes was commonly observed in patients with sepsis. Isolated bacteria showed differential sensitivity pattern against commonly used antibiotics. The majority of the isolates were resistant to several antibiotics that are usually prescribed on an empirical basis. These observations are important, especially for patient management and the development of antibiotic treatment guidelines. It is recommended that diabetic patients receive pneumococcal and influenza vaccine annually to reduce morbidity and mortality. Appropriate usage of antibiotics based on local antibiogram pattern can certainly help the clinician in reducing the burden of infections.

Keywords: antimicrobial resistance, diabetic foot ulcer, respiratory tract infection, urinary tract infection

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Authors: Vandana Bali, Manab B. Bera, Parmjit S. Panesar

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Microbial production of antimicrobials as biopreservatives is the major area of focus nowadays due to increased interest of consumers towards natural and safe preservation of ready to eat food products. The agro-industrial byproduct based medium and optimized process conditions can contribute in economical production of bacteriocins. Keeping this in view, the present investigation was carried out on agro-industrial byproducts utilization for the production of bacteriocin using Enterococcus faecium BS13 isolated from local fermented food. Different agro-industrial byproduct based carbon sources (whey, potato starch liquor, kinnow peel, deoiledrice bran and molasses), nitrogen sources (soya okra, pea pod and corn steep liquor), metal ions and surfactants were tested for optimal bacteriocin production. The effect of various process parameters such as pH, temperature, inoculum level, agitation and time were also tested on bacteriocin production. The optimized medium containing whey, supplemented with 4%corn steep liquor and polysorbate-80 displayed maximum bacteriocin activity with 2% inoculum, at pH 6.5, temperature 40oC under shaking conditions (100 rpm).

Keywords: Bacteriocin, biopreservation, corn steep liquor, Enterococcus faecium, waste utilization, whey

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Authors: Tripti Mishra, Shipra Shukla, Sanjeev Meena, , Ruchi Singh, Mahesh Pal, D. K. Upreti, Dipak Datta

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Roccella montagnei belongs to lichen family Roccelleceae growing luxuriantly along the coastal regions of India. As Roccella has been shown to be bioactive, we prepared methanolic extract and assessed its anticancer potential. The methanolic extract showed significant in vitro cytotoxic activity against four human cancer cell lines such as Colon (DLD-1, SW-620), Breast (MCF-7), Head and Neck (FaDu). This prompted us to isolate bioactive compounds through column chromatography. Two compounds Roccellic acid and Everninic acid have been isolated, out of which Everninic acid is reported for the first time. Both the compounds have been tested for in vitro cytotoxic activity in which Roccellic acid showed strong anticancer activity as compared to the Everninic acid. CDK-10 (Cyclin-dependent kinase) contributes to proliferation of cancer cells, and aberrant activity of these kinases has been reported in a wide variety of human cancers. These kinases, therefore, constitute biomarkers of proliferation and attractive pharmacological targets for the development of anticancer therapeutics. Therefore both the isolated compounds were tested for in silico molecular docking study against CDK-10 isomer enzyme to support the cytotoxic activity.

Keywords: cytotoxic activity, everninic acid, roccellic acid, R. montagnei

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Authors: A. T. Laiche, M. L. Tlil, Benine B., S. Bechoua

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The aim of this study is to isolate and select, from camel milk from El-Oued region in Algeria, a strains of lactic acid bacteria and possessing probiotic properties ; and to evaluate their potential effect on intestinal disorders in Wistar ratsmThe results relating to the selection of probiotic strains confirms that the Thermophilic streptococcus exhibits the best probiotic activity performance, with a resistance important to different degrees of pH and to bile salts, and a remarkable antibacterial activity and resistance to antibiotics compared to the other four isolated strains. In the in vivo study, diseases are induced in rats at the level of the digestive system, it was reported that the administration of Escherichia coli and castor oil caused an intestinal disorders. The microscopic observation of the histological section of the intestine showed a damaged intestinal structure and some symptoms of its irritation, including a decrease in the height of the villi and the presence of others destroyed cells, and after treatment with Streptococcus thermophilus, the microscopic observation of the cut of the histological section of the intestine showed almost complete disappearance of the mentioned symptoms, The dosage of the hematological parameters by complete blood count (CBC) is in agreement with the results of the histological sections.

Keywords: camel milk, probiotic, pathogenic bacteria, intestinal disorders, lactic acid bacteria

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Authors: Patcharakamon Nooeaid, Piyachat Chuysrinuan

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Electrospun natural polymers-based nanofibers are one of the most interesting materials used in tissue engineering and drug delivery applications. Bead-on-string nanofibers have gained considerable interest for sustained drug release. Vancomycin was used as the model drug and sodium alginate (SA)/soy protein isolated (SPI) as the polymer blend to fabricate the bead-on-string nanofibers by aqueous-based electrospinning. The bead-on-string SA/SPI nanofibers were successfully fabricated by the addition of poly(ethylene oxide) (PEO) as a co-blending polymer. SA-PEO with mass ratio of 70/30 showed the best spinnability with continuous nanofibers without the occurrence of beads. Bead structure formed with the addition of SPI and bead number increased with increasing SPI content. The electrospinning of 80/20 SA-PEO/SPI was obtained as a great promising bead-on-string nanofibers for drug loading, while the solution of 50/50 was not able to obtain continuous fibers. In vitro release tests showed that a more sustainable release profile up to 14 days with less initial burst release on day 1 could be obtained from the bead-on-string fibers than from smooth fibers with uniform diameter. In addition, vancomycin-loaded beaded fibers inhibited the growth of Staphylococcus aureus (S. aureus) bacteria. Therefore, the SA-PEO/SPI nanofibers showed the potential to be used as biomaterials for tissue engineering and drug delivery.

Keywords: bead-on-string fibers, electrospinning, drug delivery, tissue engineering

Procedia PDF Downloads 307